CA2611944A1 - Amine-containing lipids and uses thereof - Google Patents

Amine-containing lipids and uses thereof Download PDF

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Publication number
CA2611944A1
CA2611944A1 CA002611944A CA2611944A CA2611944A1 CA 2611944 A1 CA2611944 A1 CA 2611944A1 CA 002611944 A CA002611944 A CA 002611944A CA 2611944 A CA2611944 A CA 2611944A CA 2611944 A1 CA2611944 A1 CA 2611944A1
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branched
substituted
unsubstituted
unbranched
cyclic
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French (fr)
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Daniel G. Anderson
Andreas Zumbuehl
Elizaveta Sergeyevna Leshchiner
Robert S. Langer
Michael Goldberg
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Massachusetts Institute of Technology
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Massachusetts Institute Of Technology
Daniel G. Anderson
Andreas Zumbuehl
Elizaveta Sergeyevna Leshchiner
Robert S. Langer
Michael Goldberg
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Publication of CA2611944A1 publication Critical patent/CA2611944A1/en
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    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D233/00—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings
    • C07D233/54—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members
    • C07D233/56—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members with only hydrogen atoms or radicals containing only hydrogen and carbon atoms, attached to ring carbon atoms
    • C07D233/61—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members with only hydrogen atoms or radicals containing only hydrogen and carbon atoms, attached to ring carbon atoms with hydrocarbon radicals, substituted by nitrogen atoms not forming part of a nitro radical, attached to ring nitrogen atoms
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    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/16—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing nitrogen, e.g. nitro-, nitroso-, azo-compounds, nitriles, cyanates
    • A61K47/18—Amines; Amides; Ureas; Quaternary ammonium compounds; Amino acids; Oligopeptides having up to five amino acids
    • A—HUMAN NECESSITIES
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    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/22—Heterocyclic compounds, e.g. ascorbic acid, tocopherol or pyrrolidones
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    • A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
    • A61K48/0008—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy characterised by an aspect of the 'non-active' part of the composition delivered, e.g. wherein such 'non-active' part is not delivered simultaneously with the 'active' part of the composition
    • A61K48/0025—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy characterised by an aspect of the 'non-active' part of the composition delivered, e.g. wherein such 'non-active' part is not delivered simultaneously with the 'active' part of the composition wherein the non-active part clearly interacts with the delivered nucleic acid
    • A61K48/0033—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy characterised by an aspect of the 'non-active' part of the composition delivered, e.g. wherein such 'non-active' part is not delivered simultaneously with the 'active' part of the composition wherein the non-active part clearly interacts with the delivered nucleic acid the non-active part being non-polymeric
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    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
    • A61K9/16—Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction
    • A61K9/1605—Excipients; Inactive ingredients
    • A61K9/1617—Organic compounds, e.g. phospholipids, fats
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
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    • C07C227/00—Preparation of compounds containing amino and carboxyl groups bound to the same carbon skeleton
    • C07C227/04—Formation of amino groups in compounds containing carboxyl groups
    • C07C227/10—Formation of amino groups in compounds containing carboxyl groups with simultaneously increasing the number of carbon atoms in the carbon skeleton
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    • C07C229/04—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated
    • C07C229/06—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated having only one amino and one carboxyl group bound to the carbon skeleton
    • C07C229/10—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated having only one amino and one carboxyl group bound to the carbon skeleton the nitrogen atom of the amino group being further bound to acyclic carbon atoms or to carbon atoms of rings other than six-membered aromatic rings
    • C07C229/12—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated having only one amino and one carboxyl group bound to the carbon skeleton the nitrogen atom of the amino group being further bound to acyclic carbon atoms or to carbon atoms of rings other than six-membered aromatic rings to carbon atoms of acyclic carbon skeletons
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    • C07C229/04—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated
    • C07C229/06—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated having only one amino and one carboxyl group bound to the carbon skeleton
    • C07C229/10—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated having only one amino and one carboxyl group bound to the carbon skeleton the nitrogen atom of the amino group being further bound to acyclic carbon atoms or to carbon atoms of rings other than six-membered aromatic rings
    • C07C229/14—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated having only one amino and one carboxyl group bound to the carbon skeleton the nitrogen atom of the amino group being further bound to acyclic carbon atoms or to carbon atoms of rings other than six-membered aromatic rings to carbon atoms of carbon skeletons containing rings
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    • C07C229/00—Compounds containing amino and carboxyl groups bound to the same carbon skeleton
    • C07C229/02—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton
    • C07C229/04—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated
    • C07C229/06—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated having only one amino and one carboxyl group bound to the carbon skeleton
    • C07C229/10—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated having only one amino and one carboxyl group bound to the carbon skeleton the nitrogen atom of the amino group being further bound to acyclic carbon atoms or to carbon atoms of rings other than six-membered aromatic rings
    • C07C229/16—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated having only one amino and one carboxyl group bound to the carbon skeleton the nitrogen atom of the amino group being further bound to acyclic carbon atoms or to carbon atoms of rings other than six-membered aromatic rings to carbon atoms of hydrocarbon radicals substituted by amino or carboxyl groups, e.g. ethylenediamine-tetra-acetic acid, iminodiacetic acids
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    • C07C229/00—Compounds containing amino and carboxyl groups bound to the same carbon skeleton
    • C07C229/02—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton
    • C07C229/04—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated
    • C07C229/06—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated having only one amino and one carboxyl group bound to the carbon skeleton
    • C07C229/18—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated having only one amino and one carboxyl group bound to the carbon skeleton the nitrogen atom of the amino group being further bound to carbon atoms of six-membered aromatic rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C231/00—Preparation of carboxylic acid amides
    • C07C231/12—Preparation of carboxylic acid amides by reactions not involving the formation of carboxamide groups
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    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C237/00—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by amino groups
    • C07C237/02—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by amino groups having the carbon atoms of the carboxamide groups bound to acyclic carbon atoms of the carbon skeleton
    • C07C237/04—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by amino groups having the carbon atoms of the carboxamide groups bound to acyclic carbon atoms of the carbon skeleton the carbon skeleton being acyclic and saturated
    • C07C237/06—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by amino groups having the carbon atoms of the carboxamide groups bound to acyclic carbon atoms of the carbon skeleton the carbon skeleton being acyclic and saturated having the nitrogen atoms of the carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms
    • C—CHEMISTRY; METALLURGY
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    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D211/00—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings
    • C07D211/04—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D211/06—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members
    • C07D211/08—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members with hydrocarbon or substituted hydrocarbon radicals directly attached to ring carbon atoms
    • C07D211/18—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members with hydrocarbon or substituted hydrocarbon radicals directly attached to ring carbon atoms with substituted hydrocarbon radicals attached to ring carbon atoms
    • C07D211/26—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members with hydrocarbon or substituted hydrocarbon radicals directly attached to ring carbon atoms with substituted hydrocarbon radicals attached to ring carbon atoms with hydrocarbon radicals, substituted by nitrogen atoms
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    • C07D243/00—Heterocyclic compounds containing seven-membered rings having two nitrogen atoms as the only ring hetero atoms
    • C07D243/06—Heterocyclic compounds containing seven-membered rings having two nitrogen atoms as the only ring hetero atoms having the nitrogen atoms in positions 1 and 4
    • C07D243/08—Heterocyclic compounds containing seven-membered rings having two nitrogen atoms as the only ring hetero atoms having the nitrogen atoms in positions 1 and 4 not condensed with other rings
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    • C07D295/04—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with substituted hydrocarbon radicals attached to ring nitrogen atoms
    • C07D295/12—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with substituted hydrocarbon radicals attached to ring nitrogen atoms substituted by singly or doubly bound nitrogen atoms
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    • C07D317/28—Radicals substituted by nitrogen atoms
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    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/87—Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
    • C12N15/88—Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation using microencapsulation, e.g. using amphiphile liposome vesicle
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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Abstract

Nitrogen-containing lipids prepared from the conjugate addition of amines to acrylates, acrylamides, or other carbon-carbon double bonds conjugated to electron-withdrawing groups are described. Methods of preparing these lipids from commercially available starting materials are also provided. These amine-containing lipids or salts forms of these lipids are preferably biodegradable and biocompatible and may be used in a variety of drug delivery systems. Given the amino moiety of these lipids, they are particularly suited for the delivery of polynucleotides. Complexes or nanoparticles containing the inventive lipid and polynucleotide have been prepared. The inventive lipids may also be used to in preparing microparticle for drug delivery. They are particularly useful in delivering labile agents given their ability to buffer the pH of their surroundings.

Description

AMINE-CONTAINING LIPIDS AND USES THEREOF
Related Applications [0001] The present application claims priority under 35 U.S.C. 119(e) to U.S.
provisional patent applications, USSN 60/690,608, filed June 15, 2005, and USSN
60/785,176, filed March 23, 2006, each of which is incorporated herein by reference.

Government Support [0002] The work described herein was supported, in part, by grants from the National Institutes of Health (EB00244). The United States government may have certain rights in the invention.

Background of the Invention [0003] The treatment of human diseases through the application of nucleotide-based drugs such as DNA and RNA has the potential to revolutionize the medical field (Anderson Nature 392(Suppl.):25-30, 1996; Friedman Nature Med. 2:144-147, 1996; Crystal Science 270:404-410, 1995; Mulligan Science 260:926-932, 1993;
each of which is incorporated herein by reference). Thus far, the use of modified viruses as gene transfer vectors has generally represented the most clinically successful approach to gene therapy. While viral vectors are currently the most efficient gene transfer agents, concerns surrounding the overall safety of viral vectors, which include the potential for unsolicited immune responses, have resulted in parallel efforts to develop non-viral alternatives (for leading references, see: Luo et al. Nat.
Biotechnol.
18:33-37,2000; Behr Acc. Chem. Res. 26:274-278, 1993; each of which is incorporated herein by reference). Current alternatives to viral vectors include polymeric delivery systems (Zauner et al. Adv. Drug Del. Rev. 30:97-113, 1998;
Kabanov et al. Bioconjugate Chem. 6:7-20, 1995; each of which is incorporated herein by reference), liposomal formulations (Miller Angew. Chem. Int. Ed.
37:1768-1785, 1998; Hope et al. Molecular Membrane Technology 15:1-14, 1998; Deshmukh et al. New J. Chem. 21:113-124, 1997; each of which is incorporated herein by reference), and "naked" DNA injection protocols (Sanford Trends Biotechnol.
6:288-302, 1988; incorporated herein by reference). While these strategies have yet to achieve the clinical effectiveness of viral vectors, the potential safety, processing, and economic benefits offered by these methods (Anderson Nature 392(Suppl.):25-30, 1996; incorporated herein by reference) have ignited interest in the continued development of non-viral approaches to gene therapy (Boussif et al. Proc.
Natl. Acacl.
Sci. USA 92:7297-7301, 1995; Putnam et al. Macromolecules 32:3658-3662, 1999;
Lim et al. J. Am. Chem. Soc. 121:5633-5639, 1999; Gonzalez et al. Bioconjugate Chem. 10:1068-1074, 1999; Kukowska-Latallo et al. Proc. Natl. Acad. Sci. USA
93:4897-4902, 1996; Tang et al. Bioconjugate Chem. 7:703-714, 1996; Haensler et al.
Bioconjugate Chem. 4:372-379, 1993; each of which is incorporated herein by reference).
[0004] There exists a continuing need for non-toxic, biodegradable, biocompatible lipids that can be used to transfect nucleic acids and that are easily prepared efficiently and economically. Such lipids would have several uses, including the delivery of nucleic acids in gene therapy as well as in the packaging and/or delivery of diagnostic, therapeutic, and prophylactic agents.

Summary of the Invention [0005] The present invention provides novel lipids of the formula (I):
R1- \
(R5)2C-C(R6)2 ~N-R3 (R5)2C-C(R6)2 (I).
These lipids may be prepared by the addition of a primary amine to a double bond conjugated with an electron withdrawing groups such as a carbonyl moiety. Two equivalents of an a,(3-unsaturated ketone such as an acrylate are reacted with one equivalent of a primary amine to prepare the inventive lipids as shown in the scheme below:

Rl,,~,~O O,,-,~Rl O
RI ~ H2NR2 ~ N

R

These lipids typically have a hydrophobic half and a hydrophilic half. The hydrophobic portion is typically provided by fatty acid moieties attached to the acrylate, and the hydrophilic portion is provided by the esters, amines, and side chain of the amine. The fatty acid groups may be straight chain alkyl groups (CI -C30) with no substitutions. In certain embodiments, the fatty acid groups are substituted and/or branched. The amine may be protonated or alkylated thereby forming a positively charged amine. These lipids may be used in the delivery of therapeutic agents to a subject. The inventive lipids are particularly useful in delivering negatively charged agents given the tertiary amine available for protonation tlius forming a positive charge. For example, these lipids may be used to delivery DNA,RNA, or other polynucleotides to a subject or to a cell. As would be appreciated by one of skill in the art, the above reaction may result in a mixture with some lipids have one acrylate tail and other having two acrylate tails. Also, two different acrylates may be used in the reaction mixture to prepare a lipid with two different acrylate tails.
[0006] In another aspect, the invention provides lipids of the formula (II):

NiR3 A
N

Lipids of the formula (II) are prepared by the addition of a primary or secondary diamine to a double bond conjugated to an electron-withdrawing group such as a carbonyl. The lipids of formula (II) have two amines per lipid molecule as compared to the one amine per lipid molecule in the lipids of formula (I). These amines may be protonated or alkylated to form positively charged amino groups. These lipids may also be used to deliver DNA, RNA, or other polynucleotides. As with the primary amine, the acrylate tails may be the same or different. Also, the lipid may include any where from one acrylate tail to as many acrylate tails as is chemically possible.
[0007] In another aspect, the invention provides lipis of the formulae (III) or (IV):

~N A-f'u N A-IU N N A-A, N A N
(R6)2C R7 C(R6)2 (R6)2C (K6)2C
n n \i (R6)2 ~ kR5)2 (R5)2C~ ~ C(R5)2 C(R5)2 (R5)2C\

I I I I
R1 R2 (III) or R1 Rl R2 (IV).
Lipids of the formulae (III) or (IV) are prepared by the addition of primary or secondary amino groups to a double bond conjugated to an electron-withdrawing groups as as a carbonyl. The lipids of formulae (III) and (IV) have multiple amino groups per lipid molecule. In certain embodiments, the number of amino groups per lipid molecule is 3, 4, 5, 6, 7, 8, 9, or 10. These amines may be protonated or alkylated to form positively charged amino groups. The acrylate tails may all be the saine or they may be different. Any number of acrylate tails may be present on the molecule. The lipids may be used to delivery DNA, RNA, or other polynucleotides.
[0008] In one aspect of the invention, the inventive lipids are combined with an agent to form microparticles, liposomes, or micelles. The agent to be delivered by the microparticles, liposomes, or micelles may be in the form of a gas, liquid, or solid, and the agent may be a polynucleotide, protein, peptide, or small molecule.
The inventive lipids may be combined with other lipids, polymers, surfactants, cholesterol, carbohydrates, proteins, etc. to form the particles. These particles may be combined with a pharmaceutically excipient to form pharmaceutical compositions.
[0009] The invention also provides methods of making the inventive lipids.
One or more equivalents of an acrylate are allowed to react with one equivalent of a primary amine, diamine, or polyamine under suitable conditions to form a lipid of the formula (I), (II), (III), or (IV). In certain embodiments, all the amino groups of the amine are fully reacted with acrylates to form tertiary amines. In other embodiments, all the amino groups of the amine are not fully reacted with acrylate to form tertiary amines thereby resulting in primary or secondary amines in the lipid molecule.
These primary or secondary amines are left as is or may be reacted with another electrophile such as a different acrylate. As will be appreciated by one of skill in this art, reacting an amine with less than an excess of acrylate will result in a plurality of different lipid amines. Certain molecules may include a full complement of acrylate moieties while other molecules will not include a full complement of acrylates. For example, a diamine or polyamine may include only one , two, three, four, five, or six acrylate moieties off the various amino moieties of the molecule resulting in primary, secondary, and tertiary amines. In certain embodiments, it is preferred that all the amino groups not be fully functionalized. In certain embodiments, the two of the same type of acrylate are used. In other embodiments, two or more different acrylates are used. The synthesis of the lipid may be performed with or without solvent, and the synthesis may be performed at temperatures ranging from 25 C to 100 C, preferably approximately 95 C. The prepared lipids may be optionally purified. For example, the mixture of lipids may be purified to yield a lipid with a certain number of acrylate moieties. The lipids may also be alkylated using an alkyl halide (e.g., methyl iodide) or other alkylating agent.
[0010] The invention also provides libraries of lipids prepared by the inventive methods. These lipids may be prepared and/or screened using high-throughput techniques involving liquid handlers, robots, microtiter plates, computers, etc. In certain embodiments, the lipids are screened for their ability to transfect DNA, RNA, or other polynucleotides into the cell.

Definitions [0011] Definitions of specific functional groups and chemical terms are described in more detail below. For purposes of this invention, the chemical elements are identified in accordance with the Periodic Table of the Elements, CAS
version, Handbook of Chemistry and Physics, 75th Ed., inside cover, and specific functional groups are generally defined as described therein. Additionally, general principles of organic chemistry, as well as specific functional moieties and reactivity, are described in "Organic Chemistry", Thomas Sorrell, University Science Books, Sausalito:
1999, the entire contents of which are incorporated herein by reference.
[0012] Certain compounds of the present invention may exist in particular geometric or stereoisomeric forms. The present invention contemplates all such compounds, including cis- and trans-isomers, R- and S-enantiomers, diastereomers, (D)-isomers, (L)-isomers, the racemic mixtures thereof, and other mixtures thereof, as falling within the scope of the invention. Additional asymmetric carbon atoms may be present in a substituent such as an alkyl group. All such isomers, as well as mixtures thereof, are intended to be included in this invention.
[0013] Isomeric mixtures containing any of a variety of isomer ratios may be utilized in accordance with the present invention. For example, where only two isomers are combined, mixtures containing 50:50, 60:40, 70:30, 80:20, 90:10, 95:5, 96:4, 97:3, 98:2, 99:1, or 100:0 isomer ratios are all contemplated by the present invention. Those of ordinary skill in the art will readily appreciate that analogous ratios are contemplated for more complex isomer mixtures.
[0014] If, for instance, a particular enantiomer of a compound of the present invention is desired, it may be prepared by asymmetric synthesis, or by derivation with a chiral auxiliary, where the resulting diastereomeric mixture is separated and the auxiliary group cleaved to provide the pure desired enantiomers.
Alternatively, where the molecule contains a basic functional group, such as amino, or an acidic functional group, such as carboxyl, diastereomeric salts are formed with an appropriate optically-active acid or base, followed by resolution of the diastereomers thus formed by fractional crystallization or chromatographic means well known in the art, and subsequent recovery of the pure enantiomers.
[0015] One of ordinary skill in the art will appreciate that the synthetic methods, as described herein, utilize a variety of protecting groups. By the term "protecting group", as used herein, it is meant that a particular functional moiety, e.g., 0, S, or N, is temporarily blocked so that a reaction can be carried out selectively at another reactive site in a multifunctional compound. In preferred embodiments, a protecting group reacts selectively in good yield to give a protected substrate that is stable to the projected reactions; the protecting group should be selectively removable in good yield by readily available, preferably non-toxic reagents that do not attack the other functional groups; the protecting group forms an easily separable derivative (more preferably without the generation of new stereogenic centers); and the protecting group has a minimum of additional functionality to avoid further sites of reaction. As detailed herein, oxygen, sulfur, nitrogen, and carbon protecting groups may be utilized. Hydroxyl protecting groups include methyl, methoxylmethyl (MOM), methylthiomethyl (MTM), t-butylthiomethyl, (phenyldiinethylsilyl)methoxymethyl (SMOM), benzyloxymethyl (BOM), p-methoxybenzyloxymethyl (PMBM), (4-methoxyphenoxy)methyl (p-AOM), guaiacolmethyl (GUM), t-butoxymethyl, 4-pentenyloxymethyl (POM), siloxymethyl, 2-methoxyethoxymethyl (MEM), 2,2,2-trichloroethoxymethyl, bis(2-chloroethoxy)methyl, 2-(trimethylsilyl)ethoxymethyl (SEMOR), tetrahydropyranyl (THP), 3-bromotetrahydropyranyl, tetrahydrothiopyranyl, 1-methoxycyclohexyl, 4-methoxytetrahydropyranyl (MTHP), 4-methoxytetrahydrothiopyranyl, 4-methoxytetrahydrothiopyranyl S,S-dioxide, 1-[(2-chloro-4-methyl)phenyl]-4-methoxypiperidin-4-yl (CTMP), 1,4-dioxan-2-yl, tetrahydrofuranyl, tetrahydrothiofuranyl, 2,3,3a,4,5,6,7,7a-octahydro-7,8,8-trimethyl-4,7-methanobenzofuxan-2-yl, 1-ethoxyethyl, 1-(2-chloroethoxy)ethyl, 1-methyl-l-methoxyethyl, 1-methyl-l-benzyloxyethyl, 1-methyl-l-benzyloxy-2-fluoroethyl, 2,2,2-trichloroethyl, 2-trimethylsilylethyl, 2-(phenylselenyl)ethyl, t-butyl, allyl, p-chlorophenyl, p-methoxyphenyl, 2,4-dinitrophenyl, benzyl, p-methoxybenzyl, 3,4-dimethoxybenzyl, o-nitrobenzyl, p-nitrobenzyl, p-halobenzyl, 2,6-dichlorobenzyl, p-cyanobenzyl, p-phenylbenzyl, 2-picolyl, 4-picolyl, 3-methyl-2-picolyl N-oxido, diphenylmethyl, p,p'-dinitrobenzhydryl, 5-dibenzosuberyl, triphenylmethyl, a-naphthyldiphenylmethyl, p-methoxyphenyldiphenylmethyl, di(p-methoxyphenyl)phenyhnethyl, tri(p-methoxyphenyl)methyl, 4-(4'-bromophenacyloxyphenyl)diphenylmethyl, 4,4',4"-tris(4,5-dichlorophthalimidophenyl)methyl, 4,4',4"-tris(levulinoyloxyphenyl)methyl, 4,4',4"-tris(benzoyloxyphenyl)methyl, 3-(imidazol-l-y1)bis(4',4"-dimethoxyphenyl)methyl, 1,1-bis(4-methoxyphenyl)-1'-pyrenylmethyl, 9-anthryl, (9-phenyl)xanthenyl, 9-(9-phenyl-l0-oxo)anthryl, 1,3-benzodithiolan-2-yl, benzisothiazolyl S,S-dioxido, trimethylsilyl (TMS), triethylsilyl (TES), triisopropylsilyl (TIPS), dimethylisopropylsilyl (IPDMS), diethylisopropylsilyl (DEIPS), dimethylthexylsilyl, t-butyldimethylsilyl (TBDMS), t-butyldiphenylsilyl (TBDPS), tribenzylsilyl, tri p-xylylsilyl, triphenylsilyl, diphenylmethylsilyl (DPMS), t-butylmethoxyphenylsilyl (TBMPS), formate, benzoylformate, acetate, chloroacetate, dichloroacetate, trichloroacetate, trifluoroacetate, methoxyacetate, triphenylmethoxyacetate, phenoxyacetate, p-chlorophenoxyacetate, 3-phenylpropionate, 4-oxopentanoate (levulinate), 4,4-(ethylenedithio)pentanoate (levulinoyldithioacetal), pivaloate, adamantoate, crotonate, 4-methoxycrotonate, benzoate, p-phenylbenzoate, 2,4,6-trimethylbenzoate (mesitoate), alkyl methyl carbonate, 9-fluorenylmethyl carbonate (Fmoc), alkyl ethyl carbonate, alky12,2,2-trichloroethyl carbonate (Troc), 2-(trimethylsilyl)ethyl carbonate (TMSEC), 2-(phenylsulfonyl) ethyl carbonate (Psec), 2-(triphenylphosphonio) ethyl carbonate (Peoc), alkyl isobutyl carbonate, alkyl vinyl carbonate alkyl allyl carbonate, alkyl p-nitrophenyl carbonate, alkyl benzyl carbonate, alkyl p-methoxybenzyl carbonate, alky13,4-dimethoxybenzyl carbonate, alkyl o-nitrobenzyl carbonate, alkyl p-nitrobenzyl carbonate, alkyl S-benzyl thiocarbonate, 4-ethoxy- 1 -napththyl carbonate, methyl dithiocarbonate, 2-iodobenzoate, 4-azidobutyrate, 4-nitro-4-methylpentanoate, o-(dibromomethyl)benzoate, 2-formylbenzenesulfonate, 2-(methylthiomethoxy)ethyl, 4-(methylthiomethoxy)butyrate, 2-(methylthiomethoxymethyl)benzoate, 2,6-dichloro-4-methylphenoxyacetate, 2,6-dichloro-4-(1,1,3,3-tetramethylbutyl)phenoxyacetate, 2,4-bis(1,1-dimethylpropyl)phenoxyacetate, chlorodiphenylacetate, isobutyrate, monosuccinoate, (E)-2-methyl-2-butenoate, o-(methoxycarbonyl)benzoate, a-naphthoate, nitrate, alkyl N,N,N',N'-tetramethylphosphorodiamidate, alkyl N-phenylcarbamate, borate, dimethylphosphinothioyl, alky12,4-dinitrophenylsulfenate, sulfate, methanesulfonate (mesylate), benzylsulfonate, and tosylate (Ts). For protecting 1,2- or 1,3-diols, the protecting groups include methylene acetal, ethylidene acetal, 1 -t-butylethylidene ketal, 1 -phenylethylidene ketal, (4-methoxyphenyl)ethylidene acetal, 2,2,2-trichloroethylidene acetal, acetonide, cyclopentylidene ketal, cyclohexylidene ketal, cycloheptylidene ketal, benzylidene acetal, p-methoxybenzylidene acetal, 2,4-dimethoxybenzylidene ketal, 3,4-dimethoxybenzylidene acetal, 2-nitrobenzylidene acetal, methoxymethylene acetal, ethoxymethylene acetal, dimethoxymethylene ortho ester, 1-methoxyethylidene ortho ester, 1-ethoxyethylidine ortho ester, 1,2-dimethoxyethylidene ortho ester, a-methoxybenzylidene ortho ester, 1-(N,N-dimethylamino)ethylidene derivative, a-(N,N'-dimethylamino)benzylidene derivative, 2-oxacyclopentylidene ortho ester, di-t-butylsilylene group (DTBS), 1,3-(1,1,3,3-tetraisopropyldisiloxanylidene) derivative (TIPDS), tetra-t-butoxydisiloxane-1,3-diylidene derivative (TBDS), cyclic carbonates, cyclic boronates, ethyl boronate, and phenyl boronate. Amino-protecting groups include methyl carbamate, ethyl carbamante, 9-fluorenylmethyl carbamate (Fmoc), 9-(2-sulfo)fluorenylmethyl carbamate, 9-(2,7-dibromo)fluoroenylmethyl carbamate, 2,7-di-t-butyl-[9-(10,10-dioxo-10,10,10,10-tetrahydrothioxanthyl)]methyl carbamate (DBD-Tmoc), 4-methoxyphenacyl carbamate (Phenoc), 2,2,2-trichloroethyl carbamate (Troc), 2-trimethylsilylethyl carbamate (Teoc), 2-phenylethyl carbamate (hZ), 1-(1-adamantyl)-1-methylethyl carbamate (Adpoc), 1, 1 -dimethyl-2-haloethyl carbamate, 1,1-dimethyl-2,2-dibromoethyl carbamate (DB-t-BOC), 1,1-dimethyl-2,2,2-trichloroethyl carbamate (TCBOC), 1-methyl-l-(4-biphenylyl)ethyl carbamate (Bpoc), 1-(3,5-di-t-butylphenyl)-1-methylethyl carbamate (t-Bumeoc), 2-(2'-and 4'-pyridyl)ethyl carbamate (Pyoc), 2-(N,N-dicyclohexylcarboxamido)ethyl carbamate, t-butyl carbamate (BOC), 1-adamantyl carbamate (Adoc), vinyl carbamate (Voc), allyl carbamate (Alloc), 1-isopropylallyl carbamate (Ipaoc), cinnamyl carbamate (Coc), 4-nitrocimlamyl carbamate (Noc), 8-quinolyl carbamate, N-hydroxypiperidinyl carbamate, alkyldithio carbamate, benzyl carbamate (Cbz), p-methoxybenzyl carbamate (Moz), p-nitobenzyl carbamate, p-bromobenzyl carbamate, p-chlorobenzyl carbamate, 2,4-dichlorobenzyl carbamate, 4-methylsulfinylbenzyl carbamate (Msz), 9-anthrylmethyl carbamate, diphenylmethyl carbamate, 2-methylthioethyl carbamate, 2-methylsulfonylethyl carbamate, 2-(p-toluenesulfonyl)ethyl carbamate, [2-(1,3-dithianyl)]methyl carbamate (Dmoc), 4-methylthiophenyl carbamate (Mtpc), 2,4-dimethylthiophenyl carbamate (Bmpc), 2-phosphonioethyl carbamate (Peoc), 2-triphenylphosphonioisopropyl carbamate (Ppoc), 1, 1 -dimethyl-2-cyanoethyl carbamate, m-chloro-p-acyloxybenzyl carbamate, p-(dihydroxyboryl)benzyl carbamate, 5-benzisoxazolylmethyl carbamate, 2-(trifluoromethyl)-6-chromonylmethyl carbamate (Tcroc), m-nitrophenyl carbamate, 3,5-dimethoxybenzyl carbamate, o-nitrobenzyl carbamate, 3,4-dimethoxy-6-nitrobenzyl carbamate, phenyl(o-nitrophenyl)methyl carbamate, phenothiazinyl-(10)-carbonyl derivative, N'-p-toluenesulfonylaminocarbonyl derivative, N'-phenylaminothiocarbonyl derivative, t-amyl carbamate, S-benzyl thiocarbamate, p-cyanobenzyl carbamate, cyclobutyl carbamate, cyclohexyl carbamate, cyclopentyl carbamate, cyclopropylmethyl carbainate, p-decyloxybenzyl carbamate, 2,2-dimethoxycarbonylvinyl carbamate, o-(N,N-dimethylcarboxamido)benzyl carbamate, 1,1-dimethyl-3-(N,N-dimethylcarboxamido)propyl carbamate, 1, 1 -dimethylpropynyl carbamate, di(2-pyridyl)methyl carbamate, 2-furanylmethyl carbamate, 2-iodoethyl carbamate, isoborynl carbamate, isobutyl carbamate, isonicotinyl carbamate, p-(p'-methoxyphenylazo)benzyl carbamate, 1-methylcyclobutyl carbamate, 1-methylcyclohexyl carbamate, 1-methyl-l-cyclopropylmethyl carbamate, 1-methyl-l-(3,5-dimethoxyphenyl)ethyl carbamate, 1-methyl-l-(p-phenylazophenyl)ethyl carbamate, 1-methyl-l-phenylethyl carbamate, 1-methyl-l-(4-pyridyl)ethyl carbamate, phenyl carbamate, p-(phenylazo)benzyl carbamate, 2,4,6-tri-t-butylphenyl carbamate, 4-(trimethylammonium)benzyl carbamate, 2,4,6-trimethylbenzyl carbamate, formamide, acetamide, chloroacetamide, trichloroacetamide, trifluoroacetamide, phenylacetamide, 3-phenylpropanamide, picolinamide, 3-pyridylcarboxamide, N-benzoylphenylalanyl derivative, benzamide, p-phenylbenzamide, o-nitophenylacetamide, o-nitrophenoxyacetamide, acetoacetamide, (N'-dithiobenzyloxycarbonylamino)acetamide, 3-(p-hydroxyphenyl)propanamide, 3-(o-nitrophenyl)propanamide, 2-methyl-2-(o-nitrophenoxy)propanamide, 2-methyl-2-(o-phenylazophenoxy)propanamide, 4-chlorobutanamide, 3-methyl-3-nitrobutanamide, o-nitrocinnamide, N-acetylmethionine derivative, o-nitrobenzamide, o-(benzoyloxymethyl)benzamide, 4,5-diphenyl-3-oxazolin-2-one, N-phthalimide, N-dithiasuccinimide (Dts), N-2,3-diphenylmaleimide, N-2,5-dimethylpyrrole, N-1,1,4,4-tetramethyldisilylazacyclopentane adduct (STABASE), 5-substituted 1,3-dimethyl-1,3,5-triazacyclohexan-2-one, 5-substituted 1,3-dibenzyl-1,3,5-triazacyclohexan-2-one, 1-substituted 3,5-dinitro-4-pyridone, N-methylamine, N-allylamine, N-[2-(trimethylsilyl)ethoxy] methylamine (SEM), N-3-acetoxypropylamine, N-(1-isopropyl-4-nitro-2-oxo-3-pyroolin-3-yl)amine, quatemary ammonium salts, N-benzylamine, N-di(4-methoxyphenyl)methylamine, N-5-dibenzosuberylamine, N-triphenylmethylamine (Tr), N-[(4-methoxyphenyl)diphenylmethyl]amine (MMTr), N-9-phenylfluorenylamine (PhF), N-2,7-dichloro-9-fluorenylmethyleneamine,lV
ferrocenylmethylamino (Fem), N-2-picolylamino N'-oxide, N-1,1-dimethylthiomethyleneamine, N-benzylideneamine, N-p-methoxybenzylideneamine, N-diphenylmethyleneamine, N-[(2-pyridyl)mesityl]methyleneamine, N-(N')N'-dimethylaminomethylene)amine, N,N'-isopropylidenediamine, N-p-nitrobenzylideneamine, N-salicylideneamine,lV-5-chlorosalicylideneamine, N-(5-chloro-2-hydroxyphenyl)phenylmethyleneamine, N-cyclohexylideneamine, IV-(5,5-dimethyl-3-oxo-l-cyclohexenyl)amine, N-borane derivative, N-diphenylborinic acid derivative, N-[phenyl(pentacarbonylchromium- or tungsten)carbonyl]amine, N-copper chelate, N-zinc chelate, N-nitroamine, N-nitrosoamine, amine N-oxide, diphenylphosphinamide (Dpp), dimethylthiophosphinamide (Mpt), diphenylthiophosphinamide (Ppt), dialkyl phosphoramidates, dibenzyl phosphoramidate, diphenyl phosphoramidate, benzenesulfenamide, o-nitrobenzenesulfenamide (Nps), 2,4-dinitrobenzenesulfenamide, pentachlorobenzenesulfenamide, 2-nitro-4-methoxybenzenesulfenamide, triphenylmethylsulfenamide, 3-nitropyridinesulfenamide (Npys), p-toluenesulfonamide (Ts), benzenesulfonamide, 2,3,6,-trimethyl-4-methoxybenzenesulfonamide (Mtr), 2,4,6-trimethoxybenzenesulfonamide (Mtb), 2,6-dimethyl-4-methoxybenzenesulfonamide (Pme), 2,3,5,6-tetramethyl-4-methoxybenzenesulfonamide (Mte), 4-methoxybenzenesulfonamide (Mbs), 2,4,6-trimethylbenzenesulfonamide (Mts), 2,6-dimethoxy-4-methylbenzenesulfonamide (iMds), 2,2,5,7,8-pentamethylchroman-6-sulfonamide (Pmc), methanesulfonamide (Ms), (3-triinethylsilylethanesulfonamide (SES), 9-anthracenesulfonamide, 4-(4',8'-dimethoxynaphthylmethyl)benzenesulfonamide (DNMBS), benzylsulfonamide, trifluoromethylsulfonamide, and phenacylsulfonamide.. Exemplary protecting groups are detailed herein, however, it will be appreciated that the present invention is not intended to be limited to these protecting groups; rather, a variety of additional equivalent protecting groups can be readily identified using the above criteria and utilized in the method of the present invention. Additionally, a variety of protecting groups are described in Protective Groups in Organic Synthesis, Third Ed.
Greene, T.W. and Wuts, P.G., Eds., John Wiley & Sons, New York: 1999, the entire contents of which are hereby incorporated by reference.
[0016] It will be appreciated that the compounds, as described herein, may be substituted with any number of substituents or functional moieties. In general, the term "substituted" whether preceded by the term "optionally" or not, and substituents contained in formulas of this invention, refer to the replacement of hydrogen radicals in a given structure with the radical of a specified substituent. When more than one position in any given structure may be substituted with more than one substituent selected from a specified group, the substituent may be either the same or different at every position. As used herein, the term "substituted" is contemplated to include all permissible substituents of organic compounds. In a broad aspect, the perniissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and nonaromatic substituents of organic compounds. For purposes of this invention, heteroatoms such as nitrogen may have hydrogen substituents and/or any permissible substituents of organic compounds described herein which satisfy the valencies of the heteroatoms. Furthermore, this invention is not intended to be limited in any manner by the permissible substituents of organic compounds. Combinations of substituents and variables envisioned by this invention are preferably those that result in the formation of stable compounds useful in the treatment, for example, of infectious diseases or proliferative disorders. The term "stable", as used herein, preferably refers to compounds which possess stability sufficient to allow manufacture and which maintain the integrity of the compound for a sufficient period of time to be detected and preferably for a sufficient period of time to be useful for the purposes detailed herein.
[0017] The term "aliphatic", as used herein, includes both saturated and unsaturated, straight chain (i.e., unbranched), branched, acyclic, cyclic, or polycyclic aliphatic hydrocarbons, which are optionally substituted with one or more functional groups. As will be appreciated by one of ordinary skill in the art, "aliphatic" is intended herein to include, but is not limited to, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, and cycloalkynyl moieties. Thus, as used herein, the term "alkyl"
includes straight, branched and cyclic alkyl groups. An analogous convention applies to other generic terms such as "alkenyl", "alkynyl", and the like.
Furthermore, as used herein, the terms "alkyl", "alkenyl", "alkynyl", and the like encompass both substituted and unsubstituted groups. In certain embodiments, as used herein, "lower alkyl" is used to indicate those alkyl groups (cyclic, acyclic, substituted, unsubstituted, branched or unbranched) having 1-6 carbon atoms.
[0018] In certain embodiments, the alkyl, alkenyl, and alkynyl groups employed in the invention contain 1-20 aliphatic carbon atoms. In certain other embodiments, the alkyl, alkenyl, and alkynyl groups employed in the invention contain 1-10 aliphatic carbon atoms. In yet other embodiments, the alkyl, alkenyl, and alkynyl groups employed in the invention contain 1-8 aliphatic carbon atoms. In still other embodiments, the alkyl, alkenyl, and alkynyl groups employed in the invention contain 1-6 aliphatic carbon atoms. In yet other embodiments, the alkyl, alkenyl, and alkynyl groups employed in the invention contain 1-4 carbon atoms.
Illustrative aliphatic groups thus include, but are not limited to, for example, methyl, ethyl, n-propyl, isopropyl, cyclopropyl, -CH2-cyclopropyl, vinyl, allyl, n-butyl, sec-butyl, isobutyl, tert-butyl, cyclobutyl, -CH2-cyclobutyl, n-pentyl, sec-pentyl, isopentyl, tert-pentyl, cyclopentyl, -CH2-cyclopentyl, n-hexyl, sec-hexyl, cyclohexyl, -CH2-cyclohexyl moieties and the like, which again, may bear one or more substituents. Alkenyl groups include, but are not limited to, for example, ethenyl, propenyl, butenyl, 1-methyl-2-buten-l-yl, and the like. Representative alkynyl groups include, but are not limited to, ethynyl, 2-propynyl (propargyl), 1-propynyl, and the like.
[0019] The term "alkyl" as used herein refers to saturated, straight- or branched-chain hydrocarbon radicals derived from a hydrocarbon moiety containing between one and twenty carbon atoms by removal of a single hydrogen atom.
Examples of alkyl radicals include, but are not limited to, methyl, ethyl, propyl, isopropyl, n-butyl, tert-butyl, n-pentyl, neopentyl, n-hexyl, n-heptyl, n-octyl, n-decyl, n-undecyl, and dodecyl.
[0020] The term "alkenyl" denotes a monovalent group derived from a hydrocarbon moiety having at least one carbon-carbon double bond by the removal of a single hydrogen atom. Alkenyl groups include, for example, ethenyl, propenyl, butenyl, 1-methyl-2-buten-1-yl, and the like.
[0021] The term "alkynyl" as used herein refers to a monovalent group derived form a hydrocarbon having at least one carbon-carbon triple bond by the removal of a single hydrogen atoin. Representative alkynyl groups include ethynyl, 2-propynyl (propargyl), 1-propynyl, and the like.
[0022] The term "alkoxy", or "thioalkyl" as used herein refers to an alkyl group, as previously defined, attached to the parent molecule through an oxygen atom or through a sulfur atom. In certain embodiments, the alkyl, alkenyl, and alkynyl groups contain 1-20 alipahtic carbon atoms. In certain other embodiments, the alkyl, alkenyl, and alkynyl groups contain 1-10 aliphatic carbon atoms. In yet other embodiments, the alkyl, alkenyl, and alkynyl groups employed in the invention contain 1-8 aliphatic carbon atoms. In still other embodiments, the alkyl, alkenyl, and alkynyl groups contain 1-6 aliphatic carbon atoms. In yet other embodiments, the alkyl, alkenyl, and alkynyl groups contain 1-4 aliphatic carbon atoms.
Examples of alkoxy, include but are not limited to, methoxy, ethoxy, propoxy, isopropoxy, n-butoxy, tert-butoxy, neopentoxy, and n-hexoxy. Examples of thioalkyl include, but are not limited to, methylthio, ethylthio, propylthio, isopropylthio, n-butylthio, and the like.
[0023] The term "alkylamino" refers to a group having the structure -NHR', wherein R' is aliphatic, as defined herein. In certain embodiments, the aliphatic group contains 1-20 aliphatic carbon atoms. In certain other embodiments, the aliphatic group contains 1-10 aliphatic carbon atoms. In yet other embodiments, the aliphatic group employed in the invention contain 1-8 aliphatic carbon atoms. In still other embodiments, the aliphatic group contains 1-6 aliphatic carbon atoms. In yet other embodiments, the aliphatic group contains 1-4 aliphatic carbon atoms. Examples of alkylamino groups include, but are not limited to, methylamino, ethylamino, n-propylamino, iso-propylamino, cyclopropylamino, n-butylamino, tert-butylamino, neopentylamino, n-pentylamino, hexylamino, cyclohexylamino, and the like.
10024] The term "carboxylic acid" as used herein refers to a group of formula -CO2H.
10025] The term "dialkylamino" refers to a group having the structure -NRR', wherein R and R' are each an aliphatic group, as defined herein. R and R' may be the same or different in an dialkyamino moiety. In certain embodiments, the aliphatic groups contains 1-20 aliphatic carbon atoms. In certain other embodiments, the aliphatic groups contains 1-10 aliphatic carbon atoms. In yet other embodiments, the aliphatic groups employed in the invention contain 1-8 aliphatic carbon atoms.
In still other embodiments, the aliphatic groups contains 1-6 aliphatic carbon atoms.
In yet other embodiments, the aliphatic groups contains 1-4 aliphatic carbon atoms.
Examples of dialkylamino groups include, but are not limited to, dimethylamino, methyl ethylamino, diethylamino, methylpropylainino, di(n-propyl)amino, di(iso-propyl)amino, di(cyclopropyl)amino, di(n-butyl)amino, di(tert-butyl)amino, di(neopentyl)amino, di(n-pentyl)amino, di(hexyl)amino, di(cyclohexyl)amino, and the like. In certain embodiments, R and R' are linked to form a cyclic structure.
The resulting cyclic structure may be aromatic or non-aromatic. Examples of cyclic diaminoalkyl groups include, but are not limted to, aziridinyl, pyrrolidinyl, piperidinyl, morpholinyl, pyrrolyl, imidazolyl, 1,3,4-trianolyl, and tetrazolyl.
[0026] Some examples of substituents of the above-described aliphatic (and other) moieties of compounds of the invention include, but are not limited to aliphatic;
heteroaliphatic; aryl; heteroaryl; arylalkyl; heteroarylalkyl; alkoxy;
aryloxy;
heteroalkoxy; heteroaryloxy; alkylthio; arylthio; heteroalkylthio;
heteroarylthio; F; Cl;
Br; I; -OH; -NO2; -CN; -CF3; -CH2CF3; -CHC12; -CH2OH; -CH2CH2OH; -CH2NH2; -CH2SO2CH3; -C(O)RX; -CO2(Rx); -CON(RX)2; -OC(O)RX; -OCO2RX; -OCON(Rx)Z; -N(RX)2; -S(O)2RX; -NRX(CO)Rx wherein each occurrence of RX independently includes, but is not limited to, aliphatic, heteroaliphatic, aryl, heteroaryl, arylalkyl, or heteroarylalkyl, wherein any of the aliphatic, heteroaliphatic, arylalkyl, or heteroarylalkyl substituents described above and herein may be substituted or unsubstituted, branched or unbranched, cyclic or acyclic, and wherein any of the aryl or heteroaryl substituents described above and herein may be substituted or unsubstituted. Additional examples of generally applicable substituents are illustrated by the specific embodiments shown in the Examples that are described herein.
100271 In general, the terms "aryl" and "heteroaryl", as used herein, refer to stable mono- or polycyclic, heterocyclic, polycyclic, and polyheterocyclic unsaturated moieties having preferably 3-14 carbon atoms, each of which may be substituted or unsubstituted. Substituents include, but are not limited to, any of the previously mentioned substitutents, i.e., the substituents recited for aliphatic moieties, or for other moieties as disclosed herein, resulting in the formation of a stable compound. In certain embodiments of the present invention, "aryl" refers to a mono- or bicyclic carbocyclic ring system having one or two aromatic rings including, but not limited to, phenyl, naphthyl, tetrahydronaphthyl, indanyl, indenyl, and the like. In certain embodiments of the present invention, the term "heteroaryl", as used herein, refers to a cyclic aromatic radical having from five to ten ring atoms of which one ring atom is selected from S, 0, and N; zero, one, or two ring atoms are additional heteroatoms independently selected from S, 0, and N; and the remaining ring atoms are carbon, the radical being joined to the rest of the molecule via any of the ring atoms, such as, for example, pyridyl, pyrazinyl, pyrimidinyl, pyrrolyl, pyrazolyl, imidazolyl, thiazolyl, oxazolyl, isooxazolyl, thiadiazolyl,oxadiazolyl, thiophenyl, furanyl, quinolinyl, isoquinolinyl, and the like.
[00281 It will be appreciated that aryl and heteroaryl groups can be unsubstituted or substituted, wherein substitution includes replacement of one, two, three, or more of the hydrogen atoms thereon independently with any one or more of the following moieties including, but not limited to: aliphatic;
heteroaliphatic; aryl;
heteroaryl; arylalkyl; heteroarylalkyl; alkoxy; aryloxy; heteroalkoxy;
heteroaryloxy;
alkylthio; arylthio; heteroalkylthio; heteroarylthio; -F; -Cl; -Br; -I; -OH; -NO2; -CN; -CF3; -CH2CF3; -CHC12i -CHZOH; -CH2CH2OH; -CH2NH2; -CH2SO2CH3i -C(O)RX; -CO2(RY); -CON(Rx)2; -OC(O)Rx; -OCO2Rx; -OCON(RX)2; N(RX)2; -S(O)2RX; -NRX(CO)RX, wherein each occurrence of RX independently includes, but is not limited to, aliphatic, heteroaliphatic, aryl, heteroaryl, arylalkyl, or heteroarylalkyl, wherein any of the aliphatic, heteroaliphatic, arylalkyl, or heteroarylalkyl substituents described above and herein may be substituted or unsubstituted, branched or unbranched, cyclic or acyclic, and wherein any of the aryl or heteroaryl substituents described above and herein may be substituted or unsubstituted. Additional examples of generally applicable substitutents are illustrated by the specific embodiments shown in the Examples that are described herein.
[0024] The term "cycloalkyl", as used herein, refers specifically to groups having three to seven, preferably three to ten carbon atoms. Suitable cycloalkyls include, but are not limited to cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and the like, which, as in the case of other aliphatic, heteroaliphatic, or hetercyclic moieties, may optionally be substituted with substituents including, but not limited to aliphatic; heteroaliphatic; aryl; heteroaryl; arylalkyl;
heteroarylalkyl;
alkoxy; aryloxy; heteroalkoxy; heteroaryloxy; alkylthio; arylthio;
heteroalkylthio;
heteroarylthio; -F; -Cl; -Br; -I; -OH; -NO2; -CN; -CF3; -CH2CF3; -CHC12, -CHzOH; -CHZCH2OH; -CH2NH2; -CH2SO2CH3; -C(O)RX; -CO2(RX); -CON(RX)2; -OC(O)R,; -OCO2RX; -OCON(R,,)z; -N(R,,)2; -S(O)2Rx; -NRX(CO)RX, wherein each occurrence of R,t independently includes, but is not limited to, aliphatic, heteroaliphatic, aryl, heteroaryl, arylalkyl, or heteroarylalkyl, wherein any of the aliphatic, heteroaliphatic, arylalkyl, or heteroarylalkyl substituents described above and herein may be substituted or unsubstituted, branched or unbranched, cyclic or acyclic, and wherein any of the aryl or heteroaryl substituents described above and herein may be substituted or unsubstituted. Additional examples of generally applicable substitutents are illustrated by the specific embodiments shown in the Exainples that are described herein.
[0030] The term "heteroaliphatic", as used herein, refers to aliphatic moieties that contain one or more oxygen, sulfur, nitrogen, phosphorus, or silicon atoms, e.g., in place of carbon atoms. Heteroaliphatic moieties may be branched, unbranched, cyclic or acyclic and include saturated and unsaturated heterocycles such as morpholino, pyrrolidinyl, etc. In certain embodiments, heteroaliphatic moieties are substituted by independent replacement of one or more of the hydrogen atoms thereon with one or more moieties including, but not limited to aliphatic;
heteroaliphatic; aryl;
heteroaryl; arylalkyl; heteroarylalkyl; alkoxy; aryloxy; heteroalkoxy;
heteroaryloxy;
alkylthio; arylthio; heteroalkylthio; heteroarylthio; -F; -Cl; -Br; -I; -OH; -NOZ; -CN; -CF3; -CH2CF3; -CHC12; -CH2OH; -CH2CH2OH; -CH2NH2; -CH2SO2CH3; -C(O)RX; -COz(RX); -CON(RX)2; -OC(O)RX; -OCOZRx; -OCON(Rx)2; -N(Rx)2; -S(O)2Rx; -NRx(CO)Rx, wherein each occurrence of R,t independently includes, but is not limited to, aliphatic, heteroaliphatic, aryl, heteroaryl, arylalkyl, or heteroarylalkyl, wherein any of the aliphatic, heteroaliphatic, arylalkyl, or heteroarylalkyl substituents described above and herein may be substituted or unsubstituted, branched or unbranched, cyclic or acyclic, and wherein any of the aryl or heteroaryl substituents described above and herein may be substituted or unsubstituted. Additional examples of generally applicable substitutents are illustrated by the specific embodiments shown in the Examples that are described herein.
10031] The terms "halo" and "halogen" as used herein refer to an atom selected from fluorine, chlorine, bromine, and iodine.
[0032] The term "haloalkyl" denotes an alkyl group, as defined above, having one, two, or three halogen atoms attached thereto and is exemplified by such groups as chloromethyl, bromoethyl, trifluoromethyl, and the like.
[0033] The term "heterocycloalkyl" or "heterocycle", as used herein, refers to a non-aromatic 5-, 6-, or 7- membered ring or a polycyclic group, including, but not limited to a bi- or tri-cyclic group comprising fused six-membered rings having between one and three heteroatoms independently selected from oxygen, sulfur and nitrogen, wherein (i) each 5-membered ring has 0 to 1 double bonds and each 6-membered ring has 0 to 2 double bonds, (ii) the nitrogen and sulfur heteroatoms may be optionally be oxidized, (iii) the nitrogen heteroatom may optionally be quaternized, and (iv) any of the above heterocyclic rings may be fused to a benzene ring.
Representative heterocycles include, but are not limited to, pyrrolidinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, piperidinyl, piperazinyl, oxazolidinyl, isoxazolidinyl, morpholinyl, thiazolidinyl, isothiazolidinyl, and tetrahydrofuryl. In certain embodiments, a "substituted heterocycloalkyl or heterocycle" group is utilized and as used herein, refers to a heterocycloalkyl or heterocycle group, as defined above, substituted by the independent replacement of one, two or three of the hydrogen atoms thereon with but are not limited to aliphatic; heteroaliphatic;
aryl;
heteroaryl; arylalkyl; heteroarylalkyl; alkoxy; aryloxy; heteroalkoxy;
heteroaryloxy;
alkylthio; arylthio; heteroalkylthio; heteroarylthio; -F; -Cl; -Br; -I; -OH; -NO2; -CN; -CF3; -CH2CF3; -CHC12; -CH2OH; -CH2CH2OH; -CH2NH2; -CH2SO2CH3i -C(O)RX; -CO2(Rx); -CON(RX)2; -OC(O)%; -OCO2RX; -OCON(RX)2; -N(RX)2; -S(O)2RX; -NRX(CO)RX, wherein each occurrence of Rx independently includes, but is not limited to, aliphatic, heteroaliphatic, aryl, heteroaryl, arylalkyl, or heteroarylalkyl, wherein any of the aliphatic, heteroaliphatic, arylalkyl, or heteroarylalkyl substituents described above and herein may be substituted or unsubstituted, branched or unbranched, cyclic or acyclic, and wherein any of the aryl or heteroaryl substituents described above and herein may be substituted or unsubstituted. Additional examples of generally applicable substitutents are illustrated by the specific embodiments shown in the Examples which are described herein.
[0034] "Carbocycle": The term "carbocycle", as used herein, refers to an aromatic or non-aromatic ring in which each atom of the ring is a carbon atom.
[0035] "Independently selected": The term "independently selected" is used herein to indicate that the R groups can be identical or different.
[0036] "Labeled": As used herein, the term "labeled" is intended to mean that a compound has at least one element, isotope, or chemical compound attached to enable the detection of the compound. In general, labels typically fall into three classes: a) isotopic labels, which may be radioactive or heavy isotopes, including, but not limited to, 2H, 3H, 32P, 35S, 67Ga, 99i'Tc (Tc-99m), 11I In, 123I1125 I, 169Yb and 186Re; b) immune labels, which may be antibodies or antigens,which may be bound to enzyines (such as horseradish peroxidase) that produce detectable agents; and c) colored, luminescent, phosphorescent, or fluorescent dyes. It will be appreciated that the labels may be incorporated into the compound at any position that does not interfere with the biological activity or characteristic of the compound that is being detected.
In certain embodiments of the invention, photoaffi.nity labeling is utilized for the direct elucidation of intermolecular interactions in biological systems. A variety of known photophores can be employed, most relying on photoconversion of diazo compounds, azides, or diazirines to nitrenes or carbenes (See, Bayley, H., Photogenerated Reagents in Biochemistry and Molecular Biology (1983), Elsevier, Amsterdam.), the entire contents of which are hereby incorporated by reference. In certain embodiments of the invention, the photoaffinity labels employed are o-, m- and p-azidobenzoyls, substituted with one or more halogen moieties, including, but not limited to 4-azido-2,3,5,6-tetrafluorobenzoic acid.
[0037] The terms halo and halogen as used herein refer to an atom selected from fluorine, chlorine, bromine, and iodine.
[0038] The terin "heterocyclic", as used herein, refers to a non-aromatic partially unsaturated or fully saturated 3- to 10-membered ring system, which includes single rings of 3 to 8 atoms in size and bi- and tri-cyclic ring systems which may include aromatic six-membered aryl or aromatic heterocyclic groups fused to a non-aromatic ring. These heterocyclic rings include those having from one to three heteroatoms independently selected from oxygen, sulfur, and nitrogen, in which the nitrogen and sulfur heteroatoms may optionally be oxidized and the nitrogen heteroatom may optionally be quaternized.
[0039] The term "heteroaryl", as used herein, refers to a cyclic aromatic radical having from five to ten ring atoms of which one ring atom is selected from sulfur, oxygen, and nitrogen; zero, one, or two ring atoms are additional heteroatoms independently selected from sulfur, oxygen, and nitrogen; and the remaining ring atonls are carbon, the radical being joined to the rest of the molecule via any of the ring atoms, such as, for example, pyridyl, pyrazinyl, pyrimidinyl, pyrrolyl, pyrazolyl, imidazolyl, thiazolyl, oxazolyl, isooxazolyl, thiadiazolyl, oxadiazolyl, thiophenyl, furanyl, quinolinyl, isoquinolinyl, and the like.
[0040] Specific heterocyclic and aromatic heterocyclic groups that may be included in the compounds of the invention include: 3-methyl-4-(3-methylphenyl)piperazine, 3 methylpiperidine, 4-(bis-(4-fluorophenyl)methyl)piperazine, 4-(diphenylmethyl)piperazine, 4-(ethoxycarbonyl)piperazine, 4-(ethoxycarbonylmethyl)piperazine, 4-(phenylmethyl)piperazine, 4-(1-phenylethyl)piperazine, 4-(1,1-dimethylethoxycarbonyl)piperazine, 4-(2-(bis-(2-propenyl) amino)ethyl)piperazine, 4-(2-(diethylamino)ethyl)piperazine, 4-(2-chlorophenyl)piperazine, 4-(2-cyanophenyl)piperazine, 4-(2-ethoxyphenyl)piperazine, 4-(2-ethylphenyl)piperazine, 4-(2-fluorophenyl)piperazine, 4-(2-hydroxyethyl)piperazine, 4-(2-methoxyethyl)piperazine, 4-(2-methoxyphenyl)piperazine, 4-(2-methylphenyl)piperazine, 4-(2-methylthiophenyl) piperazine, 4-(2-nitrophenyl)piperazine, 4-(2-nitrophenyl)piperazine, 4-(2-phenylethyl)piperazine, 4-(2-pyridyl)piperazine, 4-(2-pyrimidinyl)piperazine, 4-(2,3-dimethylphenyl)piperazine, 4-(2,4-difluorophenyl) piperazine, 4-(2,4-dimethoxyphenyl)piperazine, 4-(2,4-dimetliylphenyl)piperazine, 4-(2,5-dimethylphenyl)piperazine, 4-(2,6-dimethylphenyl)piperazine, 4-(3 -chlorophenyl)piperazine, 4-(3 -methylphenyl)piperazine, 4-(3-trifluoromethylphenyl)piperazine, 4-(3,4-dichlorophenyl)piperazine, 4-3,4-dimethoxyphenyl)piperazine, 4-(3,4-dimethylphenyl)piperazine, 4-(3,4-methylenedioxyphenyl)piperazine, 4-(3,4,5-trimethoxyphenyl)piperazine, 4-(3,5-dichlorophenyl)piperazine, 4-(3,5-dimethoxyphenyl)piperazine, 4-(4-(phenylmethoxy)phenyl)piperazine, 4-(4-(3,1-dimethylethyl)phenylmethyl)piperazine, 4-(4-chloro-3-trifluoromethylphenyl)piperazine, 4-(4-chlorophenyl)-3-methylpiperazine, 4-(4-chlorophenyl)piperazine, 4-(4-chlorophenyl)piperazine, 4-(4-chlorophenylmethyl)piperazine, 4-(4-fluorophenyl)piperazine, 4-(4-methoxyphenyl)piperazine, 4-(4-methylphenyl)piperazine, 4-(4-nitrophenyl)piperazine, 4-(4-trifluoromethylphenyl)piperazine, 4-cyclohexylpiperazine, 4-ethylpiperazine, 4-hydroxy-4-(4-chlorophenyl)methylpiperidine, 4-hydroxy-4-phenylpiperidine, 4-hydroxypyrrolidine, 4-methylpiperazine, 4-phenylpiperazine, 4-piperidinylpiperazine, 4-(2-furanyl)carbonyl)piperazine, 4-((1,3-dioxolan-5-yl)methyl)piperazine, 6-fluoro-1,2,3,4-tetrahydro-2-methylquinoline, 1,4-diazacylcloheptane, 2,3-dihydroindolyl, 3,3-dimethylpiperidine, 4,4-ethylenedioxypiperidine, 1,2,3,4-tetrahydroisoquinoline, 1,2,3,4-tetrahydroquinoline, azacyclooctane, decahydroquinoline, piperazine, piperidine, pyrrolidine, thiomorpholine, and triazole.
[0041] The terms "substituted," whether preceded by the term "optionally" or not, and substituent, as used herein, refer to the ability, as appreciated by one skilled in this art, to change one functional group for another functional group provided that the valency of all atoms is maintained. When more than one position in any given structure may be substituted with more than one substituent selected from a specified group, the substituent may be either the same or different at every position.
The substituents may also be further substituted (e.g., an aryl group substituent may have another substituent off it, such as another aryl group, which is further substituted with fluorine at one or more positions).
[0042] The following are more general terms used throughout the present application:
[0043] "Animal": The term animal, as used herein, refers to humans as well as non-huinan animals, including, for example, mammals, birds, reptiles, amphibians, and fish. Preferably, the non-human animal is a mammal (e.g., a rodent, a mouse, a rat, a rabbit, a monkey, a dog, a cat, a primate, or a pig). An animal may be a transgenic animal.
[0044] "Associated with": When two entities are "associated with" one another as described herein, they are linked by a direct or indirect covalent or non-covalent interaction. Preferably, the association is covalent. Desirable non-covalent interactions include hydrogen bonding, van der Waals interactions, hydrophobic interactions, magnetic interactions, electrostatic interactions, etc.
[0045] "Biocompatible": The term "biocompatible", as used herein is intended to describe compounds that are not toxic to cells. Compounds are "biocompatible" if their addition to cells in vitro results in less than or equal to 20 % cell death, and their administration in vivo does not induce inflammation or other such adverse effects.

[0046] "Biodegradable": As used herein, "biodegradable" compounds are those that, when introduced into cells, are broken down by the cellular machinery or by hydrolysis into components that the cells can either reuse or dispose of without significant toxic effect on the cells (i.e., fewer than about 20 % of the cells are killed when the components are added to cells in vitro). The components preferably do not induce inflammation or other adverse effects in vivo. In certain preferred embodiments, the chemical reactions relied upon to break down the biodegradable compounds are uncatalyzed.
[0047] "Effective amount": In general, the "effective amount" of an active agent or drug delivery device refers to the amount necessary to elicit the desired biological response. As will be appreciated by those of ordinary skill in this art, the effective amount of an agent or device may vary depending on such factors as the desired biological endpoint, the agent to be delivered, the composition of the encapsulating matrix, the target tissue, etc. For example, the effective amount of microparticles containing an antigen to be delivered to immunize an individual is the amount that results in an immune response sufficient to prevent infection with an organism having the administered antigen.
[0048] "Peptide" or "protein": According to the present invention, a "peptide"
or "protein" comprises a string of at least three amino acids linked together by peptide bonds. The terms "protein" and "peptide" may be used interchangeably. Peptide may refer to an individual peptide or a collection of peptides. Inventive peptides preferably contain only natural amino acids, although non-natural amino acids (i.e., compounds that do not occur in nature but that can be incorporated into a polypeptide chain) and/or amino acid analogs as are known in the art may alternatively be employed. Also, one or more of the amino acids in an inventive peptide may be modified, for example, by the addition of a chemical entity such as a carbohydrate group, a phosphate group, a farnesyl group, an isofarnesyl group, a fatty acid group, a linker for conjugation, functionalization, or other modification, etc. In a preferred embodiment, the modifications of the peptide lead to a more stable peptide (e.g., greater half-life in vivo). These modifications may include cyclization of the peptide, the incorporation of D-amino acids, etc. None of the modifications should substantially interfere with the desired biological activity of the peptide.

[0049] "Polynucleotide" or "oligonucleotide": Polynucleotide or oligonucleotide refers to a polymer of nucleotides. Typically, a polynucleotide comprises at least three nucleotides. The polymer may include natural nucleosides (i.e., adenosine, thymidine, guanosine, cytidine, uridine, deoxyadenosine, deoxythymidine, deoxyguanosine, and deoxycytidine), nucleoside analogs (e.g., aminoadenosine, 2-thiothymidine, inosine, pyrrolo-pyrimidine, 3-methyl adenosine, C5-propynylcytidine, C5-propynyluridine, C5-bromouridine, C5-fluorouridine, C5-iodouridine, C5-methylcytidine, 7-deazaadenosine, 7-deazaguanosine, 8-oxoadenosine, 8-oxoguanosine, 0(6)-methylguanine, and 2-thiocytidine), chemically modified bases, biologically modified bases (e.g., methylated bases), intercalated bases, modified sugars (e.g., 2'-fluororibose, ribose, 2'-deoxyribose, arabinose, and hexose), or modified phosphate groups (e.g., phosphorothioates and 5' -N-phosphoramidite linkages).
[0050] "Small molecule": As used herein, the term "small molecule" refers to organic compounds, whether naturally-occurring or artificially created (e.g., via chemical synthesis) that have relatively low molecular weight and that are not proteins, polypeptides, or nucleic acids. Typically, small molecules have a molecular weight of less than about 1500 g/mol. Also, small molecules typically have multiple carbon-carbon bonds. Known naturally-occurring small molecules include, but are not limited to, penicillin, erythromycin, taxol, cyclosporin, and rapamycin.
Known synthetic small molecules include, but are not limited to, ampicillin, methicillin, sulfametlioxazole, and sulfonamides.

Brief Description of the Drawing [0051] Figure 1 shows acrylates and amines used in the synthesis of exemplary amine-containing lipids.
[0052] Figure 2 shows 1H NMR spectra of lipids LD28 (A), LD86 (B), LD87 (C), ND32 (D), ND86 (E), and ND87 (F).
[0053] Figure 3 shows the DNA transfection efficiency of several of the inventive amine-containing lipids.
[0054] Figure 4 shows the percentage of luciferase knockdown for several of the inventive lipids.

Detailed Description of Certain Preferred Embodiments of the Invention [0055] The present invention provides novel lipids and delivery systems based on the use of amino lipids. The system may be used in the pharmaceutical/drug delivery arts to delivery polynucleotides, proteins, small molecules, peptides, antigen, drugs, etc. to a patient, tissue, organ, cell, etc.
[0056] The amino lipids of the present invention provide for several different uses in the drug delivery art. The lipids with their amine-containing hydrophilic portion may be used to complex polynucleotides and thereby enhance the delivery of polynucleotide and prevent their degradation. The lipids may also be used in the formation of nanoparticles, microparticles, liposomes, and micelles containing the agent to be delivered. Preferably, the lipids are biocompatible and biodegradable, and the formed particles are also biodegradable and biocompatible and may be used to provide controlled, sustained release of the agent. These lipids and their corresponding particles may also be responsive to pH changes given that these lipids are protonated at lower pH.

Lipids [0057] The lipids of the present invention are lipids containing primary, secondary, or tertiary amines and salts thereof. In a particularly preferred embodiment, the inventive lipids are relatively non-cytotoxic. In another particularly preferred embodiment, the inventive lipids are biocompatible and biodegradable. In a particularly preferred embodiment, the lipids of the present invention have pKas in the range of 5.5 to 7.5, more preferably between 6.0 and 7Ø In another particularly preferred embodiment, the lipid may be designed to have a desired pKa between 3.0 and 9.0, more preferably between 5.0 and 8Ø The inventive lipids are particularly attractive for drug delivery for several reasons: 1) they contain amino groups for interacting with DNA, RNA, other polynucleotides, and other negatively charged agents, for buffering the pH, for causing endosomolysis, etc.; 2) they can be synthesized from commercially available starting materials; and 3) they are pH
responsive and can be engineered with a desired pKa.

[0058] In certain embodiments, the lipids of the present invention are of the formula (I):

Rj-V~
(R5)2C-C(R6)2 (R5)2C-C(R6)2 /
R2-V (I) wherein each occurrence of V is independently selected from the group consisting of C=O, C=S, S=O, and SO2;
Rl is selected from the group consisting of hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;
substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; -ORA; -C(=O)RA; -CO2RA; -CN; -SCN; -SRA; -SORA; -SO2RA; -NO2i -N3; -N(RA)2; -NHC(=0)RA; -NRAC(=0)N(RA)2; -OC(=O)ORA; -OC(=0)RA; -OC(=O)N(RA)2; -NRAC(=O)ORA; or -C(RA)3; wherein each occurrence of RA is independently a hydrogen; a protecting group; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;
substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; an acyl moiety; alkoxy; aryloxy; alkylthio; arylthio; amino, alkylamino, dialkylamino, heteroaryloxy; or heteroarylthio moiety;
R2 is selected from the group consisting of hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;
substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; -ORB; -C(=0)RB; -COZRB; -CN; -SCN; -SRB; -SORB; -SO2RB; -NO2, -N3; -N(RB)2; -NHC(=O)RB; -NRBC(=O)N(RB)2; -OC(=0)ORB; -OC(=0)RB; -OC(=O)N(RB)2; -NRBC(=O)ORB; or -C(RB)3; wherein each occurrence of RB is independently a hydrogen; a protecting group; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;
substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; an acyl moiety; alkoxy; aryloxy; alkylthio; arylthio; ainino, alkylamino, dialkylamino, heteroaryloxy; or heteroarylthio moiety;
wherein Rl and R2 may be taken together to form a cyclic structure;
R3 is selected from the group consisting of hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;
substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; -ORc; -C(=O)Rc; -CO2Rc; -CN; -SCN; -SRc; -SORc; -SO2Rc; -NOZ; -N3; -N(Rc)2; -NHC(=O)Rc; -NRcC(=O)N(Rc)2i -OC(=O)ORc; -OC(=0)Rc; -OC(=O)N(RC)2i -NRcC(=O)ORc; or -C(Rc)3i wherein each occurrence of Rc is independently a hydrogen; a protecting group; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;
substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; an acyl moiety; alkoxy; aryloxy; alkylthio; arylthio; amino, alkylamino, dialkylamino, heteroaryloxy; or heteroarylthio moiety;
each occurrence of R5 is independently selected from the group consisting of hydrogen and C 1-C6 alkyl;
each occurrence of R6 is independently selected from the group consisting of hydrogen and C1-C6 alkyl; and salts thereof.
[0059] In certain embodiments, the tertiary amine of formula (I) is protonated or alkylated to form a compound of formula (Ia):

Rl-V
(R5)2C-C (R6)2R7 (R5)2C-C(R6)2 /
R2-V (Ia) wherein Rl, R2, R3, R5, R6, and V are defined above;
R7 is hydrogen or C1-C6 aliphatic, preferably C1-C6 alkyl, more preferably hydrogen or methyl; and X is any anion. Possible anions include fluoride, chloride, bromide, iodide, sulfate, bisulfate, phosphate, nitrate, acetate, fumarate, oleate, citrate, valerate, maleate, oxalate, isonicotinate, lactate, salicylate, tartrate, tannate, pantothenate, bitartrate, ascorbate, succinate, gentisinate, gluconate, glucaronate, saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, and pamoate (i.e., 1,1'-methylene-bis-(2-hydroxy-3-naphthoate).
[0060] In certain embodiments, V is C=O. In other embodiments, V is C=S. In yet other embodiments, V is S=O. In still other embodiments, V is SOZ.
[0061] In certain embodiments, RI is hydrogen. In other embodiments, R1 is a cyclic or acyclic, substituted or unsubstituted, branched or un branched aliphatic or heteroaliphatic moiety. In certain embodiments, Rl is a substituted or unsubstituted aryl or heteroaryl moiety. Preferably, the aryl or heteroaryl moiety is a monocyclic 5-or 6-membered ring system. In certain embodiments, Rl is -ORA, -SRA, -N(RA)2, or -NHRA. In certain embodiments, Rl is -ORA. In other embodiments, Rl is -N(RA)2 or -NHRA. In certain embodiments, RA is hydrogen. In other embodiments, RA is a cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic or heteroaliphatic moiety. In certain embodiments, RA is an acyclic, substituted or unsubstituted aliphatic moiety. In certain other embodiments, RA is an acyclic, unsubstituted, unbranched aliphatic moiety, preferably C6-C30, more preferably Clo-CZO. In certain embodiments, RA is an unsubstituted, straight chain alkyl group with at least 5 carbons. In certain embodiments, RA is an unsubstituted, straight chain alkyl group, preferably C6-C30, more preferably CIO-C20. In certain embodiments, R, is -ORA, wherein RA is an unsubstituted, unbranched C9 alkyl chain. In certain embodiments, Rl is -ORA, wherein RA is an unsubstituted, unbranched CIo alkyl chain. In certain embodiments, Rl is -ORA, wherein RA is an unsubstituted, unbranched C11 alkyl chain. In certain embodiments, Rl is -ORA, wherein RA is an unsubstituted, unbranched C12 alkyl chain. In certain embodiments, Rl is -ORA, wherein RA is an unsubstituted, unbranched C13 alkyl chain. In certain embodiments, Rl is -ORA, wherein RA is an unsubstituted, unbranched C14 alkyl chain. In certain embodiments, RI is -ORA, wherein RA is an unsubstituted, unbranched C15 alkyl chain. In certain embodiments, R, is -ORA, wherein RA is an unsubstituted, unbranched C16 alkyl chain. In certain embodiments, R, is -ORA, wherein RA is an unsubstituted, unbranched C17 alkyl chain. In certain embodiments, RI is -ORA, wherein RA is an unsubstituted, unbranched C 18 alkyl chain. In certain embodiments, Rl is -ORA, wherein RA is an unsubstituted, unbranched C19 alkyl chain. In certain embodiments, Rl is -ORA, wherein RA is an unsubstituted, unbranched C20 alkyl chain. In yet other embodiments, RA is a substituted or unsubstituted aryl or heteroaryl moiety.
[0062] In certain embodiments, R2 is hydrogen. In other embodiments, R2 is a cyclic or acyclic, substituted or unsubstituted, branched or un branched aliphatic or heteroaliphatic moiety. In certain embodiments, R2 is a substituted or unsubstituted aryl or heteroaryl moiety. Preferably, the aryl or heteroaryl moiety is a monocylic 5-or 6-membered ring system. In certain embodiments, R2 is -ORB, -SRB, -N(RB)2, or -NHRB. In certain embodiments, R2 is -ORB. In other embodiments, R2 is -N(RB)2 or NHRB. In certain embodiments, RB is hydrogen. In other embodiments, RB is a cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic or heteroaliphatic moiety. In certain embodiments, RB is an acyclic, substituted or unsubstituted aliphatic moiety. In certain embodiments, RB is an unsubstituted, straight chain alkyl group with at least 5 carbons. In certain other embodiments, RB is an acyclic, unsubstituted, unbranched aliphatic moiety, preferably C6-C30, more preferably C10-C20. In certain embodiments, RB is an unsubstituted, straight chain alkyl group, preferably C6-C30, more preferably C10-C20. In certain embodiments, R2 is -ORB, wherein RB is an unsubstituted, unbranched C9 alkyl chain. In certain embodiments, R2 is -ORB, wherein RB is an unsubstituted, unbranched Clo alkyl chain. In certain embodiments, R2 is -ORB, wherein RB is an unsubstituted, unbranched C 11 alkyl chain. In certain embodiments, R2 is -ORB, wherein RB is an unsubstituted, unbranched C12 alkyl chain. In certain embodiments, R2 is -ORB, wherein RB is an unsubstituted, unbranched C13 alkyl chain. In certain embodiments, R2 is -ORB, wherein RB is an unsubstituted, unbranched C14 alkyl chain. In certain embodiments, R2 is -ORB, wherein RB is an unsubstituted, unbranched C15 alkyl chain. In certain embodiments, R2 is -ORA, wherein RB is an unsubstituted, unbranched C16 alkyl chain. In certain embodiments, R2 is -ORA, wherein RB is an unsubstituted, unbranched C17 alkyl chain. In certain embodiments, R2 is -ORA, wherein RB is an unsubstituted, unbranched C18 alkyl chain. In certain embodiments, R2 is -ORB, wherein RB is an unsubstituted, unbranched C 19 alkyl chain. In certain embodiments, R2 is -ORB, wherein RB is an unsubstituted, unbranched C20 alkyl chain. In yet other embodiments, RB is a substituted or unsubstituted aryl or heteroaryl moiety.
[0063] In certain embodiments, R3 is a cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic moiety. In other embodiments, R3 is a cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic moiety. In certain embodiments, R3 is a polyethylene glycol moiety.
In certain embodiments, R3 is an aliphatic moiety substituted with one or more hydroxyl groups. In other embodiments, R3 is an aliphatic moiety substituted with one or more amino, alkylamino, or dialkylamino groups. In certain embodiments, is a heteroaliphatic moiety. In certain embodiments, R3 is cyclic aliphatic, preferably a monocylic ring system with a 5- or 6-membered ring. In other embodiments, R3 is aryl or heteroaryl, preferably a monocyclic ring system with a 5- or 6-membered ring.
In certain embodiments, the lipids are prepared from the primary amines 1, 11, 20, 24, 25, 28, 31, 32, 36, 76, 77, 80, 86, 87, 93, 94, 95, 96, 99, or 100 shown in Figure 1. In certain other embodiments the lipids are prepared from the primary amines 31, 93, or 94 as shwon in Figure 1.
[0064] In certain embodiments, each occurrence of R5 is hydrogen. In certain embodiments, at least one occurrence of R5 is methyl and the other occurrences are hydrogen. In certain embodiments, at least two occurrences of R5 are methyl, and the other occurrences are hydrogen. In other embodiments, at least two occurrences of R5 are hydrogen.
[0065] In certain embodiments, each occurrence of R6 is hydrogen. In certain other embodiments, at least two occurrences of R6 are hydrogen. In certain embodiments, at least one occurrence of R6 is methyl, and the other occurrences are hydrogen. In certain embodiments, at least two occurrences of R6 are methyl, and the other occurrences are hydrogen.
RI- V
(R5)2C-C(R6)2 (R5)2C-C(R6)2 [0066] In certain embodiments, ~ and R2-V which Rj- \
(R5)2C-C(R6)2 are attached to N, are the same. In other embodiments, ~ and ~
(R5)2C-C(R6)2 /
R2-V which are attached to N are the same and are different than R3. In Rl- \
(R5)2C-C(R6)2 (R5)2C-"C(R6)2 yet other embodiments, , R2-V , and R3 are all different.
[0067] In certain subclasses of lipids, the lipids are of the formula:
Rq---V

wherein V, Rl, R2, and R3 are defined as above; and all occurrences of R5 and R6 are hydrogen.. In certain embodiments, Rl and R2 are the same. In other embodiments, Rl and R2 are different. In certain embodiments, V is C=0 as shown in the formula:
O
Ri O
In certain embodiments, Rl and R2 are the same. In other embodimets, Rl and R2 are different. In certain embodiments, Rl is -ORA and R2 is -ORB, as shown in the formula below:

O
Rq0 RBO
O . In certain embodiments, RA and RB are the same. In other embodiments, RA and RB are different. In certain embodiments, at least one of RA and RB is an unsubstituted, straight chain alkyl group with at least 5 carbons. In certain embodiments, both of RA and RB are an unsubstituted, straight chain alkyl group with at least 5 carbons. In certain embodiments, RA and RB are C6-C30 straight chain alkyl groups, or C21-C30 straight chain alkyl groups, preferably C9-C20 straight chain alkyl groups. In certain embodiments, RA and RB are C6-C30 straight chain alkenyl groups, or C21-C30 straight chain alkenyl groups, preferably C9-C20 straight chain alkenyl groups. In certain embodiments, RA and RB are C6-C30 straight chain alkynyl groups, or C21-C30 straight chain alkynyl groups, preferably C9-C20 straight chain alkynyl groups. In certain embodiments, when RA and RB are the same, RA and RB are not O h 1 - ~"~O"Jt"~. met y, ethyl, n propyl, or In other embodiments, when RA and RB are the same, RA and RB each comprise at least 4 carbon atoms. In other embodiments, when RA and RB are the same, RA and RB
each comprise at least 5 carbon atoms. In other embodiments, when RA and RB are the same, RA and RB each comprise at least 6 carbon atoms. In other embodiments, RA
and RB each comprise at least 4 carbon atoms. In other embodiments, RA and RB
each comprise at least 5 carbon atoms. In other embodiments, RA and RB each comprise at least 6 carbon atoms. Exemplary classes of the above formula include:

O

O
O

O
O

O
O

O
O

O
O

O
In certain embodiments, the acrylate used in the synthesis of the lipid is acrylate LD, LF, or LG in Figure 1. In certain embodiments the acrylate is acrylate LF in Figure 1. In certain embodiments the acrylate is acrylate LG in Figure 1.

O
~ RC
In certain embodiments, R3 is not O , wherein Ro is defined as above. In certain embodiments, R3 is not -CH2CH2ORc', wherein Rc' is methyl, ethyl, propyl, isopropyl, butyl, s-butyl, isobutyl, t-butyl, pentyl, cyclopentyl, hexyl, cyclohexyl, decyl, methoxymethyl, 2-methoxyethyl, 1-ethoxyethyl, 2-ethoxyethyl, (2-methoxyethoxy)methyl, 2-tetrahydrofuranyl, 2-tetrahydropyranyl, tetrahydrofurfuryl, formyl, acetyl, propionyl, butyryl, isobutyryl, pivaloyl, valeryl, methoxyacetyl, ethoxyacetyl, acetoxyacetyl, 2-formyloxyethyl, 2-acetoxyethyl, 2-oxopropyl, 2-oxobutyl, 2-oxocyclopentyl, 2-oxo-3-tetrahydrofuranyl, 2-oxo-3-tetrahydropyranyl, methoxycarbonyl, ethoxycarbonyl, and t-butoxycarbonyl. In yet other embodiments, R3 is not -CH2CH2ORc", wherein Rc" is a straight chain, branched or cyclic alkyl group of 1 to 20 carbons atoms, which may contain an ether, carbonyl, or carbonyloxy group. In yet other embodiments, R3 is not -CH2CHZORC", wherein Rc" is a straight chain, branched or cyclic alkyl group of 1 to 10 carbons atoms, which may contain an ether, carbonyl, or carbonyloxy group. In certain particular embodiments, R3 is not -CHZCH2ORC", wherein Rc" is formyl; acetyl; or methyl group.
[0068] In other embodiments, Rl is -NRA and R2 is NRB, as shown in the formula below:
O
RqHN

RBHN
O . In certain embodiments, RA and RB are the same. In other embodiments, RA and RB are different. In certain embodiments, RA and RB are C6-straight chain alkyl groups, or C21-C30 straight chain alkyl groups, preferably C9-C20 straight chain alkyl groups. In certain embodiments, RA and RB are C6-C30 straight chain alkenyl groups, or C21-C30 straight chain alkenyl groups, preferably C9-straight chain alkenyl groups. In certain embodiments, RA and RB are C6-C30 straight chain alkynyl groups, or C21-C30 straight chain alkynyl groups, preferably C9-straight chain alkynyl groups. In certain embodiments, when RA and RB are the same, O

not h 1 t 1- ro 1A~~'O~
RA and Ra are met y, e hy , n p py , or In other embodiments, when RA and RB are the same, RA and RB each comprise at least 4 carbon atoms. In other embodiments, when RA and RB are the same, RA
and RB each comprise at least 5 carbon atoms. In other embodiments, when RA and RB
are the same, RA and RB each comprise at least 6 carbon atoms. In other embodiments, RA and RB each comprise at least 4 carbon atoms. In other embodiments, RA and RB each comprise at least 5 carbon atoms. In other embodiments, RA and RB each comprise at least 6 carbon atoms. Exemplary classes of the above formula include:
O
H

H
N 4~
O
O
N
H

N
H-j O
O
H

H
N

N
H

H
N

N~
H

H
N

N~
H

H
N

In certain embodiments, the acrylate used in the synthesis of the lipid is acrylate ND, NF, NG, or NP in Figure 1. In certain embodiments the acrylate is acrylate ND
in Figure 1. In certain embodiments the acrylate is acrylate NF in Figure 1. In certain embodiments the acrylate is acrylate NP in Figure 1.

[0069] Particular exemplary compounds include:

O
OH

O N "~~N F-I

OH
O
OH
N
O N

OH
O

O N
OH
O

OH
O N N f--l OH
O
OH
N
O N

OH
O

O N
OH

O

O

N~~ OH

j"~j DN

O NN

O
~
I_ N

O N 111-1~ N ~

H
O N
OH

H O OH
~Me Me NN
O OH

O
O mMe N+
O

O

N+
U 0 Me [0070] In other subclasses of lipids, the lipids are of the formula:
Rj-V
HC~

H3C~

wherein V, Rl, and R3 are defined as above; all occurrences of R6 are hydrogen; and R5 is defined as in the formula. In certain embodiments, Rl and R2 are the same. In certain embodiments, V is C=O as shown in the formula:
O
RI

H3~ N-Rs Z
0 , preferably Rl and R2 are the same.
In certain embodiments, RI is -ORA and R2 is -ORB, as shown in the formula below:
O
RAO

RBO
0 , preferably RA and RB are the sanle. In certain embodiments, RA and RB are C6-C30 straight chain alkyl groups, preferably C9-C20 straight chain alkyl groups.

In other embodiments, Rl is -NRA and R2 is NRB, as shown in the formula below:
O
RAHN

RBHN
O , preferably RA and RB are the same. In certain embodiments, RA and RB are C6-C30 straight chain alkyl groups, preferably C9-C20 straight chain alkyl groups.
Rj- ;
(R5)2C-C(R6)2 (R5)~C-C~R6)2 [0071] In certain embodiments, ~ and R2--V in formulae (I) and (Ia) are selected from the group consisting of:

O

O

O) v O/'~~ 0 ~Ss O~
In certain embodiments, the lipids are prepared using acrylates LC, LD, LE, LF, and LG in Figure 1.

RI- V
(R5)2C-C(R6)2 (Rs)~~-~tRs)2 [0072] In certain embodiments, ~ and R2-U in formulae (I) and (Ia) are selected from the group consisting of:

O
N
H O ~Sl N
H

N)~'~
H
O
N
H
O
N' v H O

N
H
O
Nl v H

N)L"'' H O N' v '~5' H O S~
N' v H
In certain embodiments, the lipids are prepared using acrylates NC, ND, NF, NG, or NP in Figure 1. In certain embodiments, the lipids are prepared using acrylate ND.
In other embodiments, the lipids are prepared using acrylate NF.

[0073] In certain embodiments, ~ is selected from the group consisting of:

,t-\ OH jlfj\ OH ~~
N N N
OH ''L~ OH N
~ ~ O H
\ ~~

N N N /
N
~ 0 \ \
N O
N N N
OH
N N N N N

s'r!~ OH
-\-N ~.S OH
N

N N ~ O ,fs'1 0 j,S" P
\N \ \N

\
N0- \ NO \

,S'fs~ -O ,~~ 0 N O N ,~~ N
NH N N~
N "J / -\--/
NOH N OH ,~\ HO OH
N
OH

N

HO ~
N
~\ ~~ OOH
N OH N N

OH ,t", OH
-yj N
OH N N

,t"\ OH

N N NNCOH '2~ N H ''~

,~~ 0H ~\ OH
N /N
~NH '2~ OH

N-Ra [0074] In certain embodiments, ~ is selected from the group consisting of:

Sk N
.nnr Sk N
Sk N

.nnr Sk N
~Jw N
%nnr Sk N
~ .Nv N
U'\/V
N

ss~.rvtir ~N
~rvv /N'_ R3 [0075] In certain embodiments, is Sk N O OiR31 Iv n , wherein n is an interger between 0 and 10, inclusive; and R3' is hydrogen, aliphatic, heteroaliphatic, carbocyclic, heterocyclic, aryl, acyl, or heteroaryl. In certain embodiments, R3' is hydrogen, In other embodiments, R3' is C1-C6 alkyl. In yet other embodiments, R3' is acyl (e.g., acetyl).
[0076] In certain embodiments, the inventive lipid is of formula:
( i H2)nCH3 N
,,, 0 H

O

HN
I-I
(CH2)nCH3 wherein n is an integer between 5 and 20, inclusive; and m is an integer between 1 and 10, inclusive; and pharmaceutically acceptable salts thereof. In certain embodiments, n is 11. In other embodiments, n is 12.
In yet other embodiments, n is 13. In still other embodiments, n is 14. In certain embodiments, m is 1. In other embodiments, m is 2. In other embodiments, m is 3.
In other embodiments, m is 4. In other embodiments, m is 5. In other embodiments, m is 6.
[0077] In certain embodiments, the inventive lipid is of formula:
( i H2)nCH3 HN-/
,,, O H
~
wherein n is an integer between 5 and 20, inclusive; and m is an integer between 1 and 10, inclusive; and pharmaceutically acceptable salts thereof. In certain embodiments, n is 11. In other embodiments, n is 12.
In yet other embodiments, n is 13. In still other embodiments, n is 14. In certain embodiments, m is 1. In other embodiments, m is 2. In other embodiments, m is 3.
In other embodiments, m is 4. In other embodiments, m is 5. In other embodiments, mis6.
[0078] In certain embodiments, the inventive lipid is of formula:
( i H2)nCH3 m OR3' O -~-Iyf HN11-1(CH2)nCH3 wherein R3' is C1_6alkyl;
n is an integer between 5 and 20, inclusive; and m is an integer between 1 and 10, inclusive; and pharmaceutically acceptable salts thereof. In certain embodiments, R3' is methyl. In other embodiments, R3' is ethyl. In other embodiments, R3' is n-propyl., In still other embodiments, R3' is iso-propyl. In certain embodiments, n is 11. In other embodiments, n is 12. In yet other embodiments, n is 13. In still other embodiments, n is 14. In certain embodiments, m is 1. In other embodiments, m is 2. In other embodiments, m is 3. In other embodiments, m is 4. In other embodiments, m is 5. In other embodiments, m is 6.
[0079] In certain embodiments, the inventive lipid is of formula:
( i H2)nCH3 H N
m OR3'~
wherein R3' is CI_6alkyl;

n is an integer between 5 and 20, inclusive; and m is an integer between 1 and 10, inclusive; and pharmaceutically acceptable salts thereof. In certain embodiments, R3' is methyl. In other embodiments, R3' is ethyl. In other embodiments, R3' is n-propyl. In still other embodiments, R3' is iso-propyl. In certain embodiments, n is 11. In other embodiments, n is 12. In yet other embodiments, n is 13. In still other embodiments, n is 14. In certain embodiments, m is 1. In other embodiments, m is 2. In other embodiments, m is 3. In other embodiments, m is 4. In other embodiments, m is 5. In other embodiments, m is 6.
[0080] In certain embodiments, the inventive lipid is of formula:

( i H2)nCH3 N

HN
IN, (CH2)nCH3 wherein R3' is carbocyclic; heterocyclic; aryl or heteroaryl;
n is an integer between 5 and 20, inclusive; and m is an integer between 1 and 10, inclusive; and pharmaceutically acceptable salts thereof. In certain embodiments, R3' is phenyl. In other embodiments, R3' is heteroaryl. In other embodiments, R3' is aryl. In still other embodiments, R3' is histidine. In certain embodiments, n is 11. In other embodiments, n is 12. In yet other embodiments, n is 13. In still other embodiments, n is 14. In certain embodiments, m is 1. In other embodiments, m is 2. In other embodiments, m is 3.
In other embodiments, m is 4. In other embodiments, m is 5. In other embodiments, mis6.

[0081] In certain embodiments, the inventive lipid is of formula:

(i H2)nCH3 HN~
~
3, wherein R3' is carbocyclic; heterocyclic; aryl or heteroaryl;
n is an integer between 5 and 20, inclusive; and m is an integer between 1 and 10, inclusive; and pharmaceutically acceptable salts thereof. In certain embodiments, R3' is phenyl. In other embodiments, R3' is heteroaryl. In other embodiments, R3' is aryl. In still other embodiments, R3' is histidine. In certain embodiments, n is 11. In other embodiments, n is 12. In yet other embodiments, n is 13. In still other embodiments, n is 14. In certain embodiments, m is 1. In other embodiments, m is 2. In other embodiments, m is 3.
In other embodiments, m is 4. In other embodiments, in is 5. In other embodiments, mis6.
[0082] In certain embodiments, the inventive lipid is of formula:
( i H2)nCH3 N
m NH
0 N~/
HNI--,(CH2)nCH3 wherein n is an integer between 5 and 20, inclusive; and m is an integer between 1 and 10, inclusive; and pharmaceutically acceptable salts thereof. In certain embodiments, n is 11. In other embodiments, n is 12.
In yet other embodiments, n is 13. In still other embodiments, n is 14. In certain embodiments, m is 1. In other embodiments, m is 2. In other embodiments, m is 3.

In other embodiments, m is 4. In other embodiments, m is 5. In other embodiments, mis6.
[0083] In certain embodiments, the inventive lipid is of formula:
( i H2)nCH3 HN
r" N H
Nzz,/ ~
wlierein n is an integer between 5 and 20, inclusive; and m is an integer between 1 and 10, inclusive; and pharmaceutically acceptable salts thereof. In certain embodiments, n is 11. In other embodiments, n is 12.
In yet other embodiments, n is 13. In still other embodiments, n is 14. In certain embodiments, m is 1. In other embodiments, m is 2. In other embodiments, m is 3.
In other embodiments, m is 4. In other embodiments, m is 5. In other embodiments, mis6.

[0084] The present invention also provides amino lipids prepared from reacting acrylates with diamines, triamines, or polyamines. The amino moieties are completely or partially reacted with acrylate or acrylamides. Also, as would be appreciated by one of skill in this art, amino lipids with different number of acrylate or acrylamide tails will result in various isomers. These various forms of the linventive lipids are prepared individually, or the lipid is prepared as a mixture and then purified from the other forms. A single form mya be used in a composition, or a mixture of forms may be used.
[0085] The tails of the inventive amino lipids may also be the same or different.
Non-exhaustively reacted amino groups may be reacted with a second acrylate, second acrylamide, or other electrophiles to created a mixed amino lipid.
Again, various isomeric forms may be prepared and may optionally be purified.
[0086] In certain embodiments, the lipids of the present invention are of the formula (II):

Rl-V (R6)2 (R5 2C-C~N,-R3 A

N
(R5)2C-Ci R4 R2-1% (Rs)2 wherein A is selected from the group consisting of cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic; substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; and substituted or unsubstituted, branched or unbranched heteroaryl;
V is selected from the group consisting of C=O, C=S, S=O, and SO2;
Rl is selected from the group consisting of hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;
substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; -ORA; -C(=0)RA; -CO2RA; -CN; -SCN; -SRA; -SORA; -SO2RA; -NO2; -N3; -N(RA)2; -NHC(=O)RA; -NRAC(=O)N(RA)2; -OC(=0)ORA; -OC(=0)RA; -OC(=O)N(RA)2; -NRAC(=O)ORA; and -C(RA)3; wherein each occurrence of RA is independently a hydrogen; a protecting group; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branclied or unbranched heteroaliphatic;
substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; an acyl moiety; alkoxy; aryloxy; alkylthio; arylthio; amino, alkylamino, dialkylamino, heteroaryloxy; or heteroarylthio moiety;
R2 is selected from the group consisting of hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;
substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; -ORB; -C(=O)RB; -COZRB; -CN; -SCN; -SRB; -SORB; -SOZRB; -NO2; -N3; -N(RB)2; -NHC(=O)RB; -NRBC(=O)N(RB)2; -OC(=O)ORB; -OC(=O)RB; -OC(=O)N(RB)2; -NRBC(=0)ORB; or -C(RB)3; wherein each occurrence of RB is independently a hydrogen; a protecting group; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;
substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; an acyl moiety; alkoxy; aryloxy; alkylthio; arylthio; amino, alkylamino, dialkylamino, heteroaryloxy; or heteroarylthio moiety;
wherein Rl and R2 may be taken together to form a cyclic structure;
R3 is selected from the group consisting of hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;
substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; -ORo; -C(=O)Ro; -CO2Rc; -CN; -SCN; -SRo; -SORc; -SO2RC; -NO2, -N3; -N(RC)2; -NHC(=O)RC; -NRcC(=O)N(Rc)2i -OC(=O)ORc; -OC(=O)Rc; -OC(=O)N(Ro)2; -NRCC(=O)ORc; or -C(Ro)3; wherein each occurrence of RC is independently a hydrogen; a protecting group; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;
substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; an acyl moiety; alkoxy; aryloxy; alkylthio; arylthio; amino, alkylamino, dialkylamino, heteroaryloxy; or heteroarylthio moiety;
R4 is selected from the group consisting of hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;
substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; -ORD; -C(=O)RD; -COZRD; -CN; -SCN; -SRD; -SORp; -SOZRp; -NO2; -N3; -N(RD)2; -NHC(=0)RD; -NRCC(=O)N(RD)Z; -OC(=O)ORD; -OC(=O)RD; -OC(=0)N(Ro)2a -NRoC(=0)ORD; or -C(RD)3; wherein each occurrence of RD is independently a hydrogen; a protecting group; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;
substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; an acyl moiety; alkoxy; aryloxy; alkylthio; arylthio; amino, alkylamino, dialkylamino, heteroaryloxy; or heteroarylthio moiety;
wherein R3 and R4 may be taken together to form a cyclic structure;
each occurrence of R5 is independently selected from the group consisting of hydrogen and Ci-C6 alkyl;
each occurrence of R6 is independently selected from the group consisting of hydrogen and Ca-C6 alkyl; and salts thereof: In certain embodiments, the lipid is prepared using amine 95, 96, 99, 100, 103, and 109 in Figure 1. In certain embodiments, the lipid is prepared using amine 99 in Figure 1. In certain embodiments, the lipid is prepared using amine 100 in Figure 1. In certain embodiments, the lipid is prepared using acrylate ND, NF, NP, LF, and LG in Figure 1. In certain embodiments, the lipid is prepared using acrylate ND in Figure 1. In certain embodiments, the lipid is prepared using acrylate NF in Figure 1. In certain embodiments, the lipid is prepared using acrylate NP in Figure 1.
[0087) In certain embodiments, the tertiary amine of formula (II) is protonated or alkylated to form a compound of formula (IIa):
Rj- V
(R5)2C-C(R 3 5,'R

x-A

~
N' (R5)2C-C(Rs)4 R2.-.v (IIa) wherein Rl, R2, R3, R4, R5, R6, and V are defined above;

each occurrence of R7 is hydrogen or C1-C6 aliphatic, preferably C1-C6 alkyl, more preferably hydrogen or methyl;
each dashed line represents a bond or the absence of a bond, wherein when the dashed line represents a bond, the attached nitrogen is positively charged;
and X is any anion. Possible anions include fluoride, chloride, bromide', iodide, sulfate, bisulfate, phosphate, nitrate, acetate, fumarate, oleate, citrate, valerate, maleate, oxalate, isonicotinate, lactate, salicylate, tartrate, tannate, pantothenate, bitartrate, ascorbate, succinate, gentisinate, gluconate, glucaronate, saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, and pamoate (i.e., 1,1'-methylene-bis-(2-hydroxy-3-naphthoate).
In certain embodiments, both dashed lines presents bonds, and both nitrogen atoms are positively charged.
[0088] In certain embodiments, A is a cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic or heteroaliphatic group. In certain embodiments, A is a substituted or unsubstituted, branched or unbranched aliphatic group. In certain particular embodiments, A is a substituted or unsubstituted, branched or unbranched alkyl group. In certain embodiments, A is an unsubstituted, C1-C6 straight chain alkyl group. In other embodiments, A is a polyethylene group.
In yet other embodiments, A is a polyethylene glycol moiety. In certain embodiments, A, the two nitrogen atoms attached to A, R3 and R4 form a heterocyclic ring. In certain embodiments, the ring is aromatic. In other embodiments, the ring is non-aromatic. In certain embodiments, is selected from the group consisting of:

s o"/"o s ss In certain particular embodiments, is ~ . In certain embodiments, is In certain embodiments, A is n , wherein n is an interger between 0 and 10, inclusive.
[0089] In certain embodiments, V is C=O. In otller embodiments, V is C=S. In yet other embodiments, V is S=O. In still other embodiments, V is SO2.
[0090] In certain embodiments, Rl is hydrogen. In other embodiments, Rl is a cyclic or acyclic, substituted or unsubstituted, branched or un branched aliphatic or heteroaliphatic moiety. In certain embodiments, R, is a substituted or unsubstituted aryl or heteroaryl moiety. Preferably, the aryl or heteroaryl moiety is a monocylic 5-or 6-membered ring system. In certain embodiments, R, is -ORA, -SRA, -N(RA)2, or -NHRA. In certain embodiments, Rl is -ORA. In other embodiments, Rl is -N(RA)2 or NHRA. In certain embodiments, RA is hydrogen. In certain embodiments, RA is not hydrogen. In other embodiments, RA is a cyclic or acyclic, substituted or unsubstituted, branched or, unbranched aliphatic or heteroaliphatic moiety. In certain embodiments, RA is an acyclic, substituted or unsubstituted aliphatic moiety.
In certain embodiments, RA is an unsubstituted, straight chain alkyl group with at least 5 carbons. In certain other embodiments, RA is an acyclic, unsubstituted, unbranched aliphatic moiety, preferably C6-C30, more preferably C10-C20. In certain embodiments, RA is an unsubstituted, straight chain alkyl group, preferably C6-C30, more preferably CIO-C20. In certain embodiments, RI is -ORA, wherein RA is an unsubstituted, unbranched C9 alkyl chain. In certain embodiments, Rl is -ORA, wherein RA is an unsubstituted, unbranched Clo alkyl chain. In certain embodiments, Rl is -ORA, wherein RA is an unsubstituted, unbranched C11 alkyl chain. In certain embodiments, Rl is -ORA, wherein RA is an unsubstituted, unbranched C12 alkyl chain. In certain embodiments, Rl is -ORA, wherein RA is an unsubstituted, unbranched C13 alkyl chain. In certain embodiments, Rl is -ORA, wherein RA is an unsubstituted, unbranched C14 alkyl chain. In certain embodiments, Rl is -ORA, wherein RA is an unsubstituted, unbranched C15 alkyl chain. In certain embodiments, Rl is -ORA, wherein RA is an unsubstituted, unbranched C16 alkyl chain. In certain embodiments, Rl is -ORA, wherein RA is an unsubstituted, unbranched C17 alkyl chain. In certain embodiments, Rl is -ORA, wherein RA is an unsubstituted, unbranched C18 alkyl chain. In certain embodiments, Rl is -ORA, wherein RA is an unsubstituted, unbranched C19 alkyl chain. In certain embodiments, R, is -ORA, wherein RA is an unsubstituted, unbranched C20 alkyl chain. In yet other embodiments, RA is a substituted or unsubstituted aryl or heteroaryl moiety.
[0091] In certain embodiments, R2 is hydrogen. In other embodiments, R2 is a cyclic or acyclic, substituted or unsubstituted, branched or un branched aliphatic or heteroaliphatic moiety. In certain embodiments, R2 is a substituted or unsubstituted aryl or heteroaryl moiety. Preferably, the aryl or heteroaryl moiety is a monocylic 5-or 6-membered ring system. In certain embodiments, R2 is -ORB, -SRB, -N(RB)2, or -NHRB. In certain embodiments, R2 is -ORB. In other embodiments, R2 is -N(RB)2 or -NHRB. In certain embodiments, RB is hydrogen. In certain embodiments, RB is not hydrogen. In other embodiments, RB is a cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic or heteroaliphatic moiety. In certain embodiments, RB is an acyclic, substituted or unsubstituted aliphatic moiety.
In certain embodiments, RB is an unsubstituted, straight chain alkyl group with at least 5 carbons. In certain other embodiments, RB is an acyclic, unsubstituted, unbranched aliphatic moiety, preferably C6-C30, more preferably Clo-C20. In certain embodiments, RB is an unsubstituted, straight chain alkyl group, preferably C6-C30, more preferably C10-C20. In certain embodiments, R2 is -ORB, wherein RB is an unsubstituted, unbranched C9 alkyl chain. In certain embodiments, R2 is -ORB, wherein RB is an unsubstituted, unbranched C10 alkyl chain. In certain embodiments, R2 is -ORB, wherein RB is an unsubstituted, unbranched C11 alkyl chain. In certain embodiments, R2 is -ORB, wherein RB is an unsubstituted, unbranched C12 alkyl chain. In certain embodiments, R2 is -ORB, wherein RB is an unsubstituted, unbranched C13 alkyl chain. In certain embodiments, R2 is -ORB, wherein RB is an unsubstituted, unbranched C14 alkyl chain. In certain embodiments, R2 is -ORB, wherein RB is an unsubstituted, unbranched C15 alkyl chain. In certain embodiments, R2 is -ORA, wherein RB is an unsubstituted, unbranched C16 alkyl chain. In certain embodiments, R2 is -ORA, wherein RB is an unsubstituted, unbranched C17 alkyl chain. In certain embodiments, R2 is -ORA, wherein RB is an unsubstituted, unbranched C18 alkyl chain. In certain embodiments, R2 is -ORB, wherein RB is an unsubstituted, unbranched C19 alkyl chain. In certain embodiments, R2 is -ORB, wherein RB is an unsubstituted, unbranched C20 alkyl chain. In yet other embodiments, RB is a substituted or unsubstituted aryl or heteroaryl moiety.
[0092] In certain embodiments, R3 is a cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic moiety. In other embodiments, R3 is a cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic moiety. In certain embodiments, R3 is an aliphatic moiety substituted with one or more hydroxyl groups. In other embodiments, R3 is an aliphatic moiety substituted with one or more amino, alkylamino, or dialkylamino groups. In certain embodiments, R3 is C1-C6 alkyl. In certain embodiments, R3 is methyl. In certain embodiments, R3 is ethyl. In other embodiments, R3 is n-propyl. In other embodiments, R3 is iso-propyl. In certain embodiments, R3 is hydrogen. In certain embodiments, R3 is a heteroaliphatic moiety. In certain embodiments, R3 is cyclic aliphatic, preferably a monocylic ring system with a 5- or 6-membered ring. In other embodiments, R3 is aryl or heteroaryl, preferably a monocyclic ring system with a 5-or 6-membered ring. In certain embodiments, R3 is Op O / R31 , wherein n is an interger between 0 and 10, inclusive; and R3' is hydrogen, aliphatic, heteroaliphatic, carbocyclic, heterocyclic, aryl, acyl, or heteroaryl. In certain embodiments, R3' is hydrogen, In other embodiments, R3' is C1-C6 alkyl. In yet other embodiments, R3' is acyl (e.g., acetyl). In certain embodiments, R3 is . In other embodiments, R3 is [0093] In certain embodiments, R4 is a cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic moiety. In other embodiments, R4 is a cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic moiety. In certain embodiments, R4 is an aliphatic moiety substituted with one or more hydroxyl groups. In other embodiments, R4 is an aliphatic moiety substituted with one or more amino, alkylamino, or dialkylamino groups. In certain embodiments, R4 is C1-C6 alkyl. In certain embodiments, R3 is methyl. In certain embodiments, R3 is ethyl. In other embodiments, R3 is n-propyl. In other embodiments, R3 is iso-propyl. In certain embodiments, R4 is hydrogen. In certain embodiments, R4 is a heteroaliphatic moiety. In certain embodiments, R3 is cyclic aliphatic, preferably a monocylic ring system with a 5- or 6-membered ring. In other embodiments, R4 is aryl or heteroaryl, preferably a monocyclic ring system with a 5-~R4' or 6-membered ring. In certain embodiments, R4 is ~ n O , wherein n is an interger between 0 and 10, inclusive; and R4' is hydrogen, aliphatic, heteroaliphatic, carbocyclic, heterocyclic, aryl, acyl, or heteroaryl. In certain embodiments, R4' is hydrogen, In other embodiments, R4' is CI -C6 alkyl. In yet other embodiments, R4' is acyl (e.g., acetyl). In certain embodiments, R4 is . In other embodiments, R4 is .
[0094] In certain embodiments, R3 and R4 are the same. In other embodiments, R3 and R4 are different.
[0095] In certain embodiments, each occurrence of R5 is hydrogen. In certain einbodiments, at least one occurrence of R5 is methyl and the other occurrences are hydrogen. In certain embodiments, at least two occurrences of R5 are methyl, and the other occurrences are hydrogen. In other embodiments, at least two occurrences of R5 are hydrogen.
[0096] In certain embodiments, each occurrence of R6 is hydrogen. In certain other embodiments, at least two occurrences of R6 are hydrogen. In certain embodiments, at least one occurrence of R6 is methyl, and the other occurrences are hydrogen. In certain embodiments, at least two occurrences of R6 are methyl, and the other occurrences are hydrogen.
Rj- V
(R5)2C-C(R6)2 (R5)~C-C(Rs)2 [0097] In certain embodiments, ~ and R2-V which Rl- V
(R5)2C-C(R6)2 are attached to N are the same. In other embodiments, - ~ and ~
(R5)2C-C(R6)2 R2-V which are attached to N are the same and are different than R3. In Rl- V
(R5)2C-C(R6)2 (R5)~C-C(Rs)2 yet other embodiments, R2-U , and R3 are all different.
Rj- V
(R5)2C-C(R6)2 (R5);C-C(Rs)2 [0098] In certain embodiments, ~ and R2-V in formulae (II) and (IIa) are selected from the group consisting of:

O
O~~\
O

O' v ~

O

O)\'~
0 ~S
O ~S
O

In certain embodiments, the lipids are prepared using acrylates LC, LD, LE, LF, and LG in Figure 1.
RI- ~
(R5)2C-C(R6)2 (R5)~C-C~Rs)2 [0099] In certain embodiments, ~ and R2-V in formulae (II) and (IIa) are selected from the group consisting of:

N v H) O
N
H
O
N
H
O
N)'--H

N
H

N
H
O
N
H O
N
H O
N
H
In certain embodiments, the lipids are prepared using acrylates NC, ND, NF, NG, or NP in Figure 1. In certain embodiments, the lipids are prepared using acrylate ND.
In other embodiments, the lipids are prepared using acrylate NF. In other embodiments, the lipids are prepared using acrylate NP.

Rj- V
\
(R5)2C-C(R6)2 (R5)2C-C(R6)2 [00100] In certain embodiments, ~ and R2-v in RI-V
(R5)2C-C(R6)2 formulae (II) and (IIa) are the same. In other embodiments, ~ and ~
(R5)2C-C(R6)2 ~
R2-V in formulae (II) and (IIa) are different.
R3 ~Ra [00101] In certain embodiments, "'L~ is selected from the group consisting of:
vlvv N NN~~ N N
I v .vV I nr ~r ,r I r I
HON,-~ N"-"~-~OH

N~ I
N N~~ N
~v Inr Inr N0N/ N~\ N\/\
Inr =nl H H

N N/~ NN
H H H
.rvv iPr Pri~ iPr Pri i I I
~nnr ~v .nnr N N N
Iv I
V-U,,Lr R3 ~R4 N-rv A-11- N
~
[00102] In certain embodiments, ~ ,s'j is selected from the group consisting of:

~N~

~ N N
wherein R3 and R4 form a cyclic structure.

[001031 In other embodiments, the lipids of the present invention are of the formula (III):

N A-f\-,N A-n-N
(R6)2C R7 \i (Rs)2 n ~ (R5)2 (R5)2C\
V V
I I
Rl R2 (III) wherein A is selected from the group consisting of cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic; substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; and substituted or unsubstituted, branched or unbranched heteroaryl;
V is selected from the group consisting of C=O, C=S, S=O, and SO2;
n is an integer between 0 and 10, inclusive;
RI is selected from the group consisting of hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;
substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; -ORA; -C(=0)RA; -CO2RA; -CN; -SCN; -SRA; -SORA; -SO2RA; -NO2; -N3; -N(RA)2; -NHC(=0)RA; -NRAC(=O)N(RA)2; -OC(=O)ORA; -OC(=0)RA; -OC(=O)N(RA)2; -NRAC(=O)ORA; and -C(RA)3; wherein each occurrence of RA is independently a hydrogen; a protecting group; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;
substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; an acyl moiety; alkoxy; aryloxy; alkylthio; arylthio; amino, alkylamino, dialkylamino, heteroaryloxy; or heteroarylthio moiety;
R2 is selected from the group consisting of hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;
substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; -ORB; -C(=O)RB; -COZRB; -CN; -SCN; -SRB; -SORB; -SOZRB; -NO2; -N3; -N(RB)2; -NHC(=0)RB; -NRBC(=O)N(RB)2; -OC(=O)ORB; -OC(=O)RB; -OC(=O)N(RB)Z; -NRBC(=O)ORB; or -C(RB)3; wherein each occurrence of RB is independently a hydrogen; a protecting group; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;
substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; an acyl moiety; alkoxy; aryloxy; alkylthio; arylthio; amino, alkylamino, dialkylamino, heteroaryloxy; or heteroarylthio moiety;
wherein Rl and R2 may be taken together to form a cyclic structure;
R3 is selected from the group consisting of hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;
substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; -ORc; -C(=O)Ro; -CO2Ro; -CN; -SCN; -SRc; -SORc; -SO2Rc; -NOZ; -N3; -N(Rc)2; -NHC(=O)Rc; -NRCC(=O)N(Rc)Z, -OC(=O)ORc; -OC(=O)Rc; -OC(=O)N(Rc)2i -NRcC(=0)ORc; or -C(RC)3; wherein each occurrence of Rc is independently a hydrogen; a protecting group; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;
substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; an acyl moiety; alkoxy; aryloxy; alkylthio; arylthio; amino, alkylamino, dialkylamino, heteroaryloxy; or heteroarylthio moiety;
R4 is selected from the group consisting of hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;
substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; -ORD; -C(=O)RD; -CO2RD; -CN; -SCN; -SRD; -SORD; -SO2RD; -NO2, -N3; -N(RD)2; -NHC(=O)RD; -NRCC(=O)N(RD)2; -OC(=O)ORD; -OC(=0)RD; -OC(=O)N(RD)2; -NRcC(=O)ORD; or -C(RD)3; wherein each occurrence of RD is independently a hydrogen; a protecting group; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;
substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; an acyl moiety; alkoxy; aryloxy; alkylthio; arylthio; amino, alkylamino, dialkylamino, heteroaryloxy; or heteroarylthio moiety;
wherein R3 and R4 may be taken together to form a cyclic structure;
each occurrence of R5 is independently selected from the group consisting of hydrogen and C1-C6 alkyl;
each occurrence of R6 is independently selected from the group consisting of hydrogen and C1-C6 alkyl;
R7 is selected from the group consisting of hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;
substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; -ORG; -C(=O)RG; -COzRG; -CN; -SCN; -SRG; -SORG; -SOzRG; -NO2i -N3; -N(RG)2; -NHC(=O)RG; -NRGC(=O)N(RG)2; -OC(=O)ORG; -OC(=O)RG; -OC(=O)N(RG)Z; -NRGC(=O)ORG; and -C(RG)3i wherein each occurrence of RG is independently a hydrogen; a protecting group; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;
substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; an acyl moiety; alkoxy; aryloxy; alkylthio; arylthio; amino, alkylamino, dialkylamino, heteroaryloxy; or heteroarylthio moietyand salts thereof. In certain embodiments, n is 0. In other embodiments, n is 1. In still other embodiments, n is 2. In other embodimetns, n is 3. In yet other embodiments, n is 4. In other embodiments, n is 5.
In other embodiments, n is 6. In certain embodiments, the lipid is prepared using amine 98. In other embodiments, the lipid is prepared using amine 100.
[00104] In certain embodiments, the tertiary amine of formula (III) is protonated or alkylated to form a compound of formula (IIIa):
X' R8-' N A-rlj N Av'\j N/ "Re (R6)2C I/ R7 I (R6)2 n / L;(R5)2 (R5)2Q \
V V
I I
Rl R2 (IIIa) wherein RI, R2, R3, R4, R5, R6, R7, n, and V are defined above;
each occurrence of R8 is hydrogen or C1-C6 aliphatic, preferably C1-C6 alkyl, more preferably hydrogen or methyl;
each dashed line represents a bond or the absence of a bond, wherein when the dashed line represents a bond, the attached nitrogen is positively charged;
and X is any anion. Possible anions include fluoride, chloride, bromide, iodide, sulfate, bisulfate, phosphate, nitrate, acetate, fumarate, oleate, citrate, valerate, maleate, oxalate, isonicotinate, lactate, salicylate, tartrate, tannate, pantothenate, bitartrate, ascorbate, succinate, gentisinate, gluconate, glucaronate, saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, and pamoate (i.e., 1,1'-methylene-bis-(2-hydroxy-3-naphthoate).
In certain embodiments, both dashed lines presents bonds, and both nitrogen atoms are positively charged.
[00105] In certain embodiments, A is a cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic or heteroaliphatic group. In certain embodiments, A is a substituted or unsubstituted, branched or unbranched aliphatic group. In certain particular embodiments, A is a substituted or unsubstituted, branched or unbranched alkyl group. In certain embodiments, A is an unsubstituted, C1-C6 straight chain alkyl group. In other embodiments, A is a polyethylene group.
In yet other embodiments, A is a polyethylene glycol moiety. In certain embodiments, A, the two nitrogen atoms attached to A, R3 and R4 form a heterocyclic ring. In certain embodiments, the ring is aromatic. In other embodiments, the ring is non-aromatic.
[00106] In certain embodiments, V is C=O. In other embodiments, V is C=S. In yet other embodiments, V is S=O. In still other embodiments, V is SOZ.
[00107] In certain embodiments, Rl is hydrogen. In other embodiments, Rl is a cyclic or acyclic, substituted or unsubstituted, branched or un branched aliphatic or heteroaliphatic moiety. In certain embodiments, Rl is a substituted or unsubstituted aryl or heteroaryl moiety. Preferably, the aryl or heteroaryl moiety is a monocylic 5-or 6-membered ring system. In certain embodiments, Rl is -ORA, -SRA, -N(RA)2, or -NHRA. In certain embodiments, Rl is -ORA. In other embodiments, Rl is -N(RA)2 or NHRA. In certain embodiments, RA is hydrogen. In other embodiments, RA is a cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic or heteroaliphatic moiety. In certain embodiments, RA is an acyclic, substituted or unsubstituted aliphatic moiety. In certain other embodiments, RA is an acyclic, unsubstituted, unbranched aliphatic moiety, preferably C6-C30, more preferably C20. In certain embodiments, RA is an unsubstituted, straight chain alkyl group, preferably C6-C30, more preferably Clo-C20. In certain embodiments, Rl is -ORA, wherein RA is an unsubstituted, unbranched C9 alkyl chain. In certain embodiments, Rl is -ORA, wherein RA is an unsubstituted, unbranched Clo alkyl chain. In certain embodiments, Rl is -ORA, wherein RA is an unsubstituted, unbranched C11 alkyl chain. In certain embodiments, Rl is -ORA, wherein RA is an unsubstituted, unbranched C12 alkyl chain. In certain embodiments, Rl is -ORA, wherein RA is an unsubstituted, unbranched C13 alkyl chain. In certain embodiments, R, is -ORA, wherein RA is an unsubstituted, unbranched C14 alkyl chain. In certain embodiments, Rl is -ORA, wherein RA is an unsubstituted, unbranched C15 alkyl chain. In certain embodiments, Rl is -ORA, wherein RA is an unsubstituted, unbranched C16 alkyl chain. In certain embodiments, Rl is -ORA, wherein RA is an unsubstituted, unbranched C17 alkyl chain. In certain embodiments, Rl is -ORA, wherein RA is an unsubstituted, unbranched C18 alkyl chain. In certain embodiments, Rl is -ORA, wherein RA is an unsubstituted, unbranched C19 alkyl chain. In certain embodiments, Rl is -ORA, wherein RA is an unsubstituted, unbranched C20 alkyl chain. In yet other embodiments, RA is a substituted or unsubstituted aryl or heteroaryl moiety.
[00108] In certain embodiments, R2 is hydrogen. In other embodiments, R2 is a cyclic or acyclic, substituted or unsubstituted, branched or un branched aliphatic or heteroaliphatic moiety. In certain embodiments, R2 is a substituted or unsubstituted aryl or heteroaryl moiety. Preferably, the aryl or heteroaryl moiety is a monocylic 5-or 6-membered ring system. In certain embodiments, R2 is -ORB, -SRB, -N(RB)2, or -NHRB. In certain embodiments, R2 is -ORB. In other embodiments, R2 is -N(RB)2 or -NHRB. In certain embodiments, RB is hydrogen. In other embodiments, RB is a cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic or heteroaliphatic moiety. In certain embodiments, RB is an acyclic, substituted or unsubstituted aliphatic moiety. In certain other embodiments, RB is an acyclic, unsubstituted, unbranched aliphatic moiety, preferably C6-C30, more preferably C1o-C20. In certain embodiments, RB is an unsubstituted, straight chain alkyl group, preferably C6-C30, more preferably C10-Ca0. In certain embodiments, R2 is -ORB, wherein RB is an unsubstituted, unbranched C9 alkyl chain. In certain embodiments, R2 is -ORB, wherein RB is an unsubstituted, unbranched C 10 alkyl chain. In certain embodiments, R2 is -ORB, wherein RB is an unsubstituted, unbranched CI 1 alkyl chain. In certain embodiments, R2 is -ORB, wherein RB is an unsubstituted, unbranched C12 alkyl chain. In certain embodiments, R2 is -ORB, wherein RB is an unsubstituted, unbranched C13 alkyl chain. In certain embodiments, R2 is -ORB, wherein RB is an unsubstituted, unbranched C14 alkyl chain. In certain embodiments, R2 is -ORB, wherein RB is an unsubstituted, unbranched C15 alkyl chain. In certain embodiments, R2 is -ORA, wherein RB is an unsubstituted, unbranched C16 alkyl chain. In certain embodiments, R2 is -ORA, wherein RB is an unsubstituted, unbranched C17 alkyl chain. In certain embodiments, R2 is -ORA, wherein RB is an unsubstituted, unbranched C18 alkyl chain. In certain embodiments, R2 is -ORB, wherein RB is an unsubstituted, unbranched C19 alkyl chain. In certain embodiments, R2 is -ORB, wherein RB is an unsubstituted, unbranched.C20 alkyl chain. In yet other embodiments, RB is a substituted or unsubstituted aryl or heteroaryl moiety.
[001091 In certain embodiments, R3 is hydrogen. In certain embodiments, R3 is a cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic moiety. In other embodiments, R3 is a cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic moiety. In certain embodiments, R3 is an aliphatic moiety substituted with one or more hydroxyl groups. In other embodiments, R3 is an aliphatic moiety substituted with one or more amino, alkylamino, or dialkylamino groups. In certain embodiments, R3 is C1-C6 alkyl.
In certain embodiments, R3 is hydrogen. In certain embodiments, R3 is a heteroaliphatic moiety. In certain embodiments, R3 is cyclic aliphatic, preferably a monocylic ring system with a 5- or 6-membered ring. In other embodiments, R3 is aryl or heteroaryl, preferably a monocyclic ring system with a 5- or 6-membered ring. In certain R, ~ ~
(R5)2C-C(R6)2 (R5)~C-C(Rs)2 embodiments, R3 is or R2-V , wherein Rl, R2, R5, R6, and V are defined as above.
[00110] In other embodimets, R4 is hydrogen. In certain embodiments, R4 is a cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic moiety. In other embodiments, R4 is a cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic moiety. In certain embodiments, R4 is an aliphatic moiety substituted with one or more hydroxyl groups. In other embodiments, R4 is an aliphatic moiety substituted with one or more ainino, alkylamino, or dialkylamino groups. In certain embodiments, R4 is C1-C6 alkyl.
In certain embodiments, R4 is hydrogen. In certain embodiments, R4 is a heteroaliphatic moiety. In certain embodiments, R3 is cyclic aliphatic, preferably a monocylic ring system with a 5- or 6-membered ring. In other embodiments, R4 is aryl or heteroaryl, preferably a monocyclic ring system with a 5- or 6-membered ring. In certain RI- ~
(R5)2C-C(R6)2 (R5)~C-C~R6)2 embodiments, R4 is or R2-V , wherein Rl, R2, R5, R6, and V are defined as above.
[00111] In certain embodiments, R3 and R4 are the same. In other embodiments, R3 and R4 are different. In certain embodiments, both R3 and R4 are hydrogen.
In certain embodiments, only one of R3 and R4 is hydrogen. In certain embodiments, Rl- ~
(R5)2C-C(R6)2 (R5)~C-C~Rs)z both R3 and R4 are or R2-U , wherein RI, R2, R5, R6, and V are defined as above. In certain embodiments, one of R3 and R4 is Rj- V /
(R5)2C-C(R6)2 (R5)~C-~~R6)2 ~ or R2-v , wherein Rl, R2, R5, R6, and V are defined as above; and the other is hydrogen. In certain embodiments, both R3 and R4 are Rl-CO

wherein Rlis as defined above. In certain embodiments, one of R3 Rl-CO

and R4 is , wherein Rl is defined as above; and the other is hydrogen.
In certain embodiments, each occurrence of R5 is hydrogen. In certain embodiments, at least one occurrence of R5 is methyl and the other occurrences are hydrogen. In certain embodiments, at least two occurrences of R5 are methyl, and the other occurrences are hydrogen. In other embodiments, at least two occurrences of R5 are hydrogen.
In certain embodiments, each occurrence of R6 is liydrogen. In certain other embodiments, at least two occurrences of R6 are hydrogen. In certain embodiments, at least one occurrence of R6 is methyl, and the other occurrences are hydrogen. In certain embodiments, at least two occurrences of R6 are methyl, and the other occurrences are hydrogen.

RIr (R5)2C-C(R6)2 (R5)2C-C(R6)2 [001121 In certain embodiments, R7 is or R2-V
wherein Rl, R2, R5, R6, and V are defined as above. In certain embodiments, R7, RI-V
(R5)2 -C(R6)2 (R5)~C-C(R6)2 ~, and R2-V are the same. In other embodiments, R7, Rl- V
(R5)2C-C(R6)2 (R5)~C-C(R6)2 ~, and R2-V are different. In certain embodiments, R7 RI-V
(R5)2C-C(R6)2 (R5)~C-C(R6)2 and ~ are the same. In other embodiments, R7 and R2-V
are the same. In certain embodiments, all R7 are the same.
Rl- V
(R5)2C-C(R6)2 (R5)fC'C(R6)2 [00113] In certain embod'unents, ~ and R2-V which Rj- V
(R5)2C-C(R6)2 are attached to N are the same. In other embodiments, ~ and ~
(R5)2C._'C(R6)2 ~
R2-'V which are attached to N are the same and are different than R3 or RI- ~
(R5)2C-C(R6)2 (R5)2 CT C(R6)2 R4. In yet other embodiments, , R2-V , R3, and R4 are all different. In certain embodiments, R3 and R4 are the same. In other embodiments, R3 and R4 are different.
RI \
(R5)2C-C(R6)2 )2 (R5)2C-C(R6 [00114] In certain embodiments, \ and R2-'V in formulae (III) and (IIIa) are selected from the group consisting of:

0 ~S

0) \~~FF

''II
0 Jv~\Ss 0' v ' S' In certain embodiments, the lipids are prepared using acrylates LC, LD, LE, LF, and LG in Figure 1.
Rl- ~
(R5)2C-C(R6)2 (R5)~C-C~R6)2 [00115] In certain embodiments, ~ and R2-v in formulae (III) and (IIIa) are- selected from the group consisting of:

N)L""~

NJ"~
H O

N
H

N' v H
O
N
H O ~S
N
H ~Ss NJ"' H

N' v ' N' v '~,S' H 0 S~
N" v H
In certain embodiments, the lipids are prepared using acrylates NC, ND, NF, NG, and NP in Figure 1. In certain embodiments, the lipids are prepared using acrylate ND.
In other embodiments, the lipids are prepared using acrylate NF.

A~
[00116] In certain embodiments, is selected from the group consisting of:

o~"o ~ s ~
s s-,'~ s ~-U- A., In certain particular embodiments, is G . In certain particular embodiments, is and n is 0, 1, 2, 3, 4, 5, or 6. In certain particular embodiments, is ~ and n is 2.
~
~-,P q ~n, In certain embodiments, is ~. In certain embodiments, is and n is 0, 1, 2, 3, 4, 5, or 6. In certain embodiments, is and n is 2.

[00117] In certain embodiments, the lipid is of the formula (IV):

N N
R7 x N R7 ly x wherein each occurrence of x is an integer between 1 and 10, inclusive; preferably, between 1 and 6, inclusive;

y is an integer between 0 and 10, inclusive; preferably, between 0 and 6, inclusive;
each occurrence of R7 is hydrogen; substituted or unsubstituted, branched or unbranched aliphatic; substituted or unsubstituted, branched or unbranched heteroaliphatic; substituted or unsubstituted aryl; substituted or unsubstituted Rl-CO

heteroaryl; or Rl is selected from the group consisting of hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;
substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; -ORA; -C(=O)RA; -CO2RA; -CN; -SCN; -SRA; -SORA; -SO2RA; -NO2; -N3; -N(RA)2; -NHC(=0)RA; -NRAC(=O)N(RA)2; -OC(=0)ORA; -OC(=0)RA; -OC(=O)N(RA)2; -NRAC(=0)ORA; and -C(RA)3; wherein each occurrence of RA is independently a hydrogen; a protecting group; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;
substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; an acyl moiety; alkoxy; aryloxy; alkylthio; arylthio; amino, alkylamino, dialkylamino, heteroaryloxy; or heteroarylthio moiety; and salts thereof. In certain embodiments, x is 1, 2, 3, 4, or 5. In certain particular embodiments, x is 1. In other particular embodiments, x is 2. In certain embodiments, y is 0. In certain embodiments, y is 1.
In other embodiments, y is 2. In yet other embodiments, y is 3. In still other embodiments, y is 4. In certain embodiments, R, is -ORA. In other embodiments, R, is -NHRA. In certain embodiments, at least one R, is C1-C20 alkyl. In certain Rl-CO
embodiments, all R7 are of the formula . In certain embodiments, at least one R7 is branched or unbranched, substituted or unsubstituted aliphatic. In certain embodiments, at least one R7 is C1-CZO alkyl. In certain embodiments, at least one R7 is C1-C12 alkyl. In certain embodiments, at least one R7 is branched or unbranched, substituted or unsubstituted heteroaliphatic. In certain embodiments, at ' least one R7 is CZ2" k 0 , wherein k is an interger between 0 and 10, inclusive, and R7' is hydrogen or Ca_6alkyl. In certain embodiments, at least one R7 NHa R~ is G . In other embodiments, at least one R7 is . In other embodiments, at least one R7 is a hydrogen. In other embodiments, at least two R7 are each hydrogen. In still other embodiments, at least three R7 are each hydrogen.
In still fiurthe embodiments, at least four R7 are each hydrogen.
[00118] In certain embodiments, each R7 in formulae (IV) is independently selected from the group consisting of hydrogen and O)L"v i ss O' v Ol v OAv O
[00119] In certain embodiments, each R7 in formulae (IV) is independently selected from the group consisting of hydrogen and O
N

N S.S'r H O

N
H
O
N
H
O
N )11S.SS
H O

N
O
N )~S,SS
H O

H O
H O

N S,S's H

[00120] In certain embodiments, the lipid is of the forinula (V), (VI), or (VII):
I8 I$ I$
~N'NR7 R7 x ~N~~/('N, R, R$ 11-1 N\\~N'-~R7 R7 x I I ( R7 (V), R7 (VI), or R8 (VII) wherein x is an integer between 1 and 10, inclusive; preferably, between 1 and 6, inclusive; more preferably, between 1 and 3, inclusive;
RI-CO
each occurrence of R7 is hydrogen or R, is selected from the group consisting of hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;
substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; -ORA; -C(=O)RA; -CO2RA; -CN; -SCN; -SRA; -SORA; -SOzRA; -NOZ; -N3; -N(RA)2; -NHC(=O)RA; -NRAC(=0)N(RA)2; -OC(=O)ORA; -OC(=0)RA; -OC(=O)N(RA)2; -NRAC(=0)ORA; and -C(RA)3; wlierein each occurrence of RA is independently a hydrogen; a protecting group; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;
substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; an acyl moiety; alkoxy; aryloxy; alkylthio; arylthio; amino, alkylamino, dialkylamino, heteroaryloxy; or heteroarylthio moiety;
each occurrence of R8 is independently hydrogen, C1-C6 alkyl, hydroxy-C1-C6-alkyl; or '~ k 0 , wherein k is an interger between 0 and 10, inclusive, and R8' is hydrogen or C1_6alkyl; and salts thereof. In certain embodiments, x is 1, 2, 3, 4, or 5. In certain particular embodiments, x is 1. In other particular embodiments, x is 2. In other embodiments, x is 3. In certain embodiments, Rl is -ORA. In other embodiments, Rl is NHRA. In certain embodiments, all R7 are of the Rj-CO

formula . In other embodiments, at least one R7 is a hydrogen. In other embodiments, at least two R7 are each hydrogen. In still other embodiments, at least three R7 are each hydrogen. In still furthe embodiments, at least four R7 are each hydrogen. In certain embodiments, all R$ are the same. In certain particular embodiments, R8 is hydrogen. In certain embodiments, R8 is methyl. In other embodiments, R8 is ethyl. In yet other embodiments, R8 is hydroxymethyl. In still other embodiments, R8 is hydroxyethyl.
[00121] In certain embodiments, each R7 in formula (V), (VI), or (VII) is independently selected from the group consisting of hydrogen and o o) o o) v o o) \
rSs [00122] In certain embodiments, each R7 in fonnula (V), (VI), or (VII) is independently selected from the group consisting of hydrogen and N~

N
H
O
N
H

N
H

~S
N" v H
O
N' v H

N
H

N
H O
H O
N
H
[00123] Exemplary compounds of the formulae (V), (VI), and (VII) are of the fomzula:

I e IR 7 IE t IE t M
N N~ N N

Me Me i7 R7 /N\~~ R7 M e ~N N,. R7 ' Et R7 (iH2)20H R7 N N
EtN R7 (CH2)2 \R7 0~

i7 (CH2)20H R7 R7N'~~ i ~ iN

(CH2)20N
In certain embodiments, the lipid is one of the formulae:
( i H2)nCH3 HNN "~~ N /(CH2)nCH3 H
O '~Yf HN
N-I
(CH2)nCH3 0 i N H
(CH2)nCH3 ( i H2)nCH3 N
"~ N H

O )-f HNNI-I
(CH2)nCH3 0 i N H

(CH2)nCH3 ( i H2)nCH3 ( i H2)nCH3 HN 0 HN O

HN
NH
N
"*'~ N H 2 (CH2)nCH3 (CH2)nCH3 or ( i H2)nCH3 H
NH2~

wherein n is an integer ranging from 1 to 15, inclusive; prefererably, n is an integer ranging from 6 to 12, inclusive, or 1 to 6, inclusive. In certain embodiments, n is 1, 2, 3, 4, 5, 6, 7, 9, 9, 10, 11, 12, 13, 14, or 15. In certain particular embodiments, n is 10, 11, or 12. In certain embodiments, n is 11. In other emboidments, n is 10. In certain embodiments, each n is independently an integer ranging from 1 to 15, inclusive. In other embodiments, all n are the same integer. In certain embodiments, one n is different from the other n in the compound.
[001241 In other embodiments, the coinpound is of one of the formulae:

( i (CH2)nCH3 N --~ N 0 ,,(CH2)nCH3 O

(CH2)nCH3 0 i (CH2)nCH3 ( i (CH2)nCH3 N
"'~ NH

0 --Tf (CH2)nCH3 0 i (CH2)nCH3 (iH2)nCH3 ( i (CH2)nCH3 0 0 H N
N H
N
~~NH2 (CH2)nCH3 (CH2)nCH3 or ( i H2)nCH3 H~~
NH2, wherein n is an integer ranging from 1 to 15, inclusive; prefererably, n is an integer ranging from 6 to 12, inclusive, or 1 to 6, inclusive. In certain embodiments, n is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15. In certain particular embodiments, n is 10, 11, or 12. In certain embodiments, n is 11. In other emboidments, n is 10. In certain embodiments, each n is independently an integer ranging from 1 to 15, inclusive. In other embodiments, all n are the same integer. In certain embodiments, one n is different from the other n in the compound.
[00125] In certain embodiments, the lipid is of one of the formulae:

(CH2)nCH3 HN N N N
H
(CH2)nCH3 I I
(CH2)nCH3 (CH2)nCH3 H N N N H
I
(CH2)nCH3 I I
((;H2)nCH3 (CH2)nCH3 H i N N NH2 (CH2)nCH3 I
(CH2)nCH3 0 iN H 0 iN H 0 iN H
(CH2)nCH3 (CH2)nCH3 or (CH2)nCH3, wherein n is an integer ranging from 1 to 15, inclusive; prefererably, n is an integer ranging from 6 to 12, inclusive, or 1 to 6, inclusive. In certain embodiments, n is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15. In certain particular embodiments, n is 10, 11, or 12. In certain embodiments, n is 11. In other emboidments, n is 10. In certain embodiments, each n is independently an integer ranging from 1 to 15, inclusive. In other embodiments, all n are the same integer. In certain embodiments, one n is different from the other n in the compound.
[00126] In certain embodiments, the lipid is of one of the formulae:

N N "~Ao /(CH2)nCH3 O

(CH2)nCH3 (CH2)nCH3 (CH2)nCH3 I
(CH2)nCH3 (CH2)nCH3 (CH2)nCH3 0 )L~"~ i N NH2 (CH2)nCH3 (CH2)nCH3 0 i 0 i 0 i (CH2)nCH3 (CH2)nCH3 or (CH2)nCH3, wherein n is an integer ranging from 1 to 15, inclusive; prefererably, n is an integer ranging from 6 to 12, inclusive, or 1 to 6, inclusive. In certain embodiments, n is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15. In certain particular embodiments, n is 10, 11, or 12. In certain embodiments, n is 11. In other emboidments, n is 10. In certain embodiments, each n is independently an integer ranging from 1 to 15, inclusive. In other embodiments, all n are the same integer. In certain embodiments, one n is different from the other n in the compound.

[00127] In certain embodiments, the lipid is of one of the formulae:

( i H2)nCH3 ( i H2)nCH3 N~~ /(CH2)nCH3 N N H
0 ZZTf HN
INI
NH
(CH2)nCH3 0 I NH 0 I

(CH2)nCH3 (CH2)nCH3 ( i (CH2)nCH3 N~~ N N N /(CH2)nCH3 N

HN
N H
(CH2)nCH3 0 I N H 0 I
(CH2)nCH3 (CH2)nCH3 ( i (CH2)nCH3 ( i (CH2)nCH3 N~~ N /(CH2)nCH3 H N H
0 lzz~Yf HN
INI
(CH2)nCH3 0 i N H
(CH2)nCH3 ( i (CH2)nCH3 H
N-"~ N-~ N-"~ NH
O --Zllf HN
NH
(CH2)nCH3 0 I N H 0 I
(CH2)nCH3 (CH2)nCH3 ( i (CH2)nCH3 N~~ N N /(CH2)nCH3 H N H

HN
(CH2)nCH3 0 i N H
(CH2)nCH3 ( i (CH2)nCH3 HN~\ N N -~A /(CH2)nCHs H

I I
(CH2)nCH3 (CH2)nCH3 ( i H2)nCH3 H
N--'~ NNN N~(CH2)nCH3 H H
O lz)lf HN

(CH2)nCH3 0 i N H
(CH2)nCH3 H H
N---~ NNNI-II(CH2)nCH3 H H
O _ZYf HN

(CH2)nCH3 0 i NH
(CH2)nCH3 ( i H2)nCH3 H
HNNNNH

I I
(CH2)nCH3 (CH2)nCH3 ( i (CH2)nCH3 ( i (CH2)nCH3 N-"-~HNNH2 0 --ZTf HNN" (CH2)nCH3 ( i (CH2)nCH3 H
N--'~ NN--'~NH2 0 'Yf HN
(CH2)nCH3 0 i N H
((;H2)nCH3 ( i H2)nCH3 N~~~ NH2 I
(CH2)nCH3 ( i H2)nCH3 H
HNN~~NH2 I
(CH2)nCH3 ( i H2)nCH3 H
N-'-~HNNH2 O _Yf HNI--, (CH2)nCH3 H H
N--'~ NNNH
H

0 ZYf HN
(CH2)nCH3 0 i N H
(CH2)nCH3 H H
N--'~HN~~NH2 N O -Z~f HN
NI-I(CH2)nCH3 or H

N --'~NH2 I
(CH2)nCH3 wherein n is an integer ranging from 1 to 15, inclusive; prefererably, n is an integer ranging from 6 to 12, inclusive, or 1 to 6, inclusive. In certain embodiments, n is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15. In certain particular embodiments, n is 10, 11, or 12. In certain embodiments, n is 11. In other emboidments, n is 10. In certain embodiments, each n is independently an integer ranging from 1 to 15, inclusive. In other embodiments, all n are the same integer. In certain embodiments, one n is different from the other n in the compound.

[00128] In certain embodiments, the lipid is of one of the formulae:
( i H2)nCH3 ( i H2)nCH3 ,-(CH2)nCH3 N N N O
N
O ':ZYf (CH2)nCH3 0 i 0 0 0 (CH2)nCH3 (CH2)nCH3 ( i (CH2)nCH3 H
N--'~ NNN 0,,(CH2)nCH3 O ~rf (CH2)nCH3 0 i 0 i (CH2)nCH3 (CH2)nCH3 ( i (CH2)nCH3 ( i (CH2)nCH3 N--'~ N-~ N--'~ N 0,(CH2)nCH3 H

(CH2)nCH3 0 i (CH2)nCH3 ( i (CH2)nCH3 H
NNN--'-~N H

(CH2)nCH3 0 i 0 i (CH2)nCH3 (CH2)nCH3 ( i H2)nCH3 /(CH2)nCH3 N N N N O
H
O _Yf (CH2)nCH3 0 0 (CH2)nCH3 ( i H2)nCH3 O O

H
/(CH2)nCH3 HN--'~ N N N "-~ O

(CH2)nCH3 (CH2)nCH3 ( i H2)nCH3 H
/(CH2)nCH3 NN N N'-~~ J~ 0 H

O
1~11 (CH2)nCH3 0 0 (CH2)nCH3 H H
N--'~ NN N -,~A 0~ (C H2)nC H 3 H

(CH2)nCH3 0 0 (CH2)nCH3 ( i (CH2)nCH3 H
HN~'~ NNNH

(CH2)nCH3 (CH2)nCH3 ( i (CH2)nCH3 ( i (CH2)nCH3 NHN~~NH2 1-.(CH2)nCH3 ( i H2)nCH3 H
N--'~ NNNH2 0 _Yf (CH2)nCH3 0 i (CH2)nCH3 ( i H2)nCH3 (CH2)nCH3 ( i H2)nCH3 H
HN"~~ NN""~ NH2 (CH2)nCH3 ( i H2)nCH3 O O
H
N"~~HN~~NH2 N~'(CH2)nCH3 H H
NN
H

O -z:zTf O
INI
(CH2)nCH3 0 0 (CH2)nCH3 H H
NHN N~~NH2 0 4zzzl-f N-1(CH2)nCH3 or H

( (CH2)nCH3 wherein n is an integer ranging from 1 to 15, inclusive; prefererably, n is an integer ranging from 6 to 12, inclusive, or 1 to 6, inclusive. In certain embodiments, n is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15. In certain particular embodiments, n is 10, 11, or 12. In certain embodiments, n is 11. In other emboidments, n is 10. In certain embodiments, each n is independently an integer ranging from 1 to 15, inclusive. In other embodiments, all n are the same integer. In certain embodiments, one n is different from the other n in the compound.

[00129) In another aspect of the invention, the lipid or composition of lipids of the invention is lipid or composition prepared by reacting an amine of one of the formula (1-117):
OH
1 O.,--,,NH2 24 NO,/,NH2 77 C_ ~ V NH2 NHZ N
6cOY~ 25 HO}NH2 I\ NH2 ~O~NH2 HO NH 79 \N~
7 0 26 HO,~6H 80 N,_,,,,NH2 -O'--NH2 28 HO,-~~NH2 81 N"~NH2 ~pNH 2 31 HO~~O~~NH2 82 CN-~-NM2 13 -O 32 H0-/"-/'NHZ 86 HO"~N_j-NH2 NH2 33 HO~NH2 HO' 2 87 HO~N~

NH2 34 HO NH2 I~

HO'~NH2 36 HO NN2 90 0\---/N----NH2 21 Y,,NH 91 0 NNH2 OH 93 ~ N~'--NM2 N~~N, H H 94 HN~NH2 61 -~N~~N~ ~N

H H
62 N-,,,N,,<
H 96 H~~NHz 98 H2N'/-N--,N"---NH2 H
64 H H 99 H2N~iNH~

H
75 NH2 103 HOl/'N-,,,N~~OH
H
H
76 CN -,,_,NH2 109 H2N,,~N '-'-"OH

95 ~H~UNHz 112 H2N/~/~H,~'NH:

96 'H-~NHz 113 HaN~/~N~~NH~
H
98 HZN"~N-,,N=/~NH= 114 HYN"~N-,,iNHz H H
99 HzNi,~NH% NH2 100 HzN'-'-'-'NHz N-~

109 HaN-_N---~OH HZ
/NH2 116 õoNHp HZN~~N J( 110 ~ Hz NHz 111 HzN,/-N~iNNH2 H H
with an acrylate of formula:
O O
LA NA
H
O O
LB NB NA~
H

H

LD p~0 ND
H

LE pJL,,o- NE
H

LF ~ NF H
p 0 LG NG H

0 NP LG.2 H

LH NH H~
In certain embodiments, one equivalent of amine is reacted with one equivalent of acrylate. In certain embdiments, one equivalent of amine is reacted with one, two, three, four, five, six, or more equivalents of acrylate. In certain embodiments, the amount of acrylate is limiting to prevent the functionalization of all amino groups.
The resulting lipid or lipid composition in these instances contain secondary amino groups or primary amino groups. Lipids having secondary amines are particular useful in certain instances. In certain embodiments, amine-containing lipids that have not been fully functionalize are futher reacted with another electrophile (e.g., an acrylate, acrylamide, alkylating agent, acylating agent, etc.). Such further functionalization of the amines of the lipid results in lipids with different tails. One, two, three, four, five, or more tails may be different from the other tails of the lipid.
[00130] In certain embodiments, the amine and acrylate are reacted together neat. In other embodiments, the reaction is done in a solvent (e.g., THF, CH2CI2, MeOH, EtOH, CHC13, hexanes, toluene, benzene, CC14, glyme, diethyl ether, etc.). In certain embodiments, the reaction mixture is heated. In a particularly preferred embodiment, the reaction mixture is heated to temperature ranging from 50-150 C.
In another particularly preferred embodiment, the reaction mixture is heated to approximately 95 C. The reaction may also be catalyzed. For example, the reaction may be catalyzed by the addition of an acid, base, or metal. The reaction may be allowed to proceed for hours, days, or weeks. In certain embodiments, the reaction is allowed to proceed for 1-7 days, preferably 7 days. The resulting composition may be used with or without purification. In certain embodiments, the lipids are subsequently subjected to an alkylation step (e.g., reaction with methyl iodide) to form quanternary amine salts. Optionally, various salt forms of the lipids may be prepared. In certain embodiments, the salts are pharmaceutically acceptable salts.

[00131] In certain embodiments, the lipid is prepared by reacting amine 98 with acrylate NC to form lipid NC98. In certain embodiments, the lipid NC98 is of one of the formulae below:

( i H2)10CH3 (CH2)10CH3 H2)10CH3 NNN -"-~ N,(CH2)10CH3 H

HN
(CH2)10CH3 0 I NH 0 I
N H
(CH2)10CH3 (CH2)10CH3 (i H2)1oCH3 H
NN~~=NN "-~ N/(CH2)1oCH3 H

HN
N H
(CH2)1oCH3 0 I N H 0 I

(CH2)10L;H3 (CH2)10CH3 ( i H2)1oCH3 ( i H2)1oCH3 HNN/(CH2)1oCH3 H
0 ~-f HN
(CH2)1oCH3 0 NH
I
(CH2)10CH3 ( i H2)IoCH3 H
N-"~ N_--~ N--'~ N H

HNN-I
N H
(CH2)10CH3 0 I NH 0 I

(CH2)10CH3 (CH2)10CH3 ( i H2)10CH3 H
NNNN N.-(CH2)10CH3 H H
O zYf HNINI
(CH2)IoCH3 0 i N H
(CH2)10CH3 ( i H2)IoCHs H
HNNN N,(CH2)10CH3 H

I I
(CH2)10CH3 (CH2)10CH3 ( i H2)10CH3 H
NNNN N/(CH2)10CH3 H H
O

HN
I--, (CH2)10CH3 0 i H
(CH2)10CH3 H H --~ N--'~ NNN N/(CH2)1oCH3 H H
O

HN
(CH2)10CH3 0 i N H
(CH2)10CH3 ( i H2)10CH3 H
HNNNNH
0 iN H 0 iN H

(CH2)10CH3 (CH2)1oCH3 ( i H2)10CH3 (CH2)10CH3 H2)10CH3 N--'~H~~~N~~NH2 0 -Z)lf HN
N'-,(CH2)1oCH3 ( i H2)10CH3 H
NNN-'-~NH2 O ::ZI-f HNI--, (CH2)1oCH3 0 i N H
(CH2)1oCH3 ( i H2)10CH3 N~~ NH2 I
(CH2)10CH3 ( i H2)loCH3 H
HN_-~ N-"~NH2 I
(CH2)IoCH3 ( i H2)1oCHa H
N--'~HNNH2 HN
NN(CH2)10CH3 H H
NNNNH
H

O '~Tf (CH2)10CH3 0 i N H

(CH2)loCH3 H H
H
NN--'~NH2 O _Yf HN
I*-.(CH2)loCH3 or H

I
(CH2)10CH3 In other embodiments, the lipid is a composition of one or more of the above lipids.
[00132] In certain embodiments, the lipid is prepared by reacting amine 99 with acrylate NC to form lipid NC99. In certain embodiments, the lipid NC99 is of one of the formulae below:

( i H2)10CH3 N N N/(CH2)1oCH3 H
0 ZYf HN
(CH2)10CH3 0 i N H
(CH2)10CH3 (CH2)10CH3 H2)10CH3 N
'*'~ N H

O -~-f HN
NI-I
(CH2)10CH3 0 i N H
(CH2)1oCH3 (CH2)10CH3 H2)100H3 (CH2)10CH3 H2)10CH3 HN 0 H
NH
N
~"~NH2 HN I
(CH2)10CH3 (L;H2)10CH3 or ( i H2)1o0H3 HN
NH2, In other embodiments, the lipid is a composition of one or more of the above lipids.
[00133] In certain embodiments, the lipid is prepared by reacting amine 100 with acrylate NC to form lipid NC 100. In certain embodiments, the lipid NC 100 is of one of the formulae below:

HN "'~ N N N,(CH2)10CH3 I H
(CH2)10CH3 I I
(CH2)1oCH3 (CH2)10CH3 H N N N H
(CH2)10CH3 I I
(CH2)10CH3 (CH2)10CH3 H i N )111~ N NH2 (CH2)1oCH3 ( (CH2)1oCH3 I ( I
(CH2)10CH3 (CH2)1oeH3 or (CH2)10CH3.
In other embodiments, the lipid is a composition of one or more of the above lipids.
[00134] In certain embodiments, the lipid is prepared by reacting amine 20 with acrylate ND to form lipid ND20. In certain embodiments, the lipid ND20 is of one of the formulae below:
(iH2)11CH3 ( i H2)11CH3 HNIN, (CH2)11CH3 or HN OH

In other embodiments, the lipid is a composition of one or more of the above lipids.
[00135] In certain embodiments, the lipid is prepared by reacting amine 24 with acrylate ND to form lipid ND24. In certain embodiments, the lipid ND24 is of one of the formulae below:

(i H2)1,CH3 OH (CH2)11CH3 H2)11CH3 N OH HN O

0 zYf OH
HN~(CH2)11CH3 or HN OH

In other embodiments, the lipid is a composition of one or more of the above lipids.
[00136] In certain embodiments, the lipid is prepared by reacting amine 25 with acrylate ND to form lipid ND25. In certain embodiments, the lipid ND25 is of one of the formulae below:

( i H2)11CH3 ( i H2)11CH3 OH

N
OH
O HN
OH

(CH2)11CH3 or OH.

In other embodiments, the lipid is a composition of one or more of the above lipids.
[00137] In certain embodiments, the lipid is prepared by reacting amine 28 with acrylate ND to form lipid ND28. In certain embodiments, the lipid ND28 is of one of the formulae below:

( i H2)11CH3 ( i H2)11CH3 0 -Tf (CH2)11CH3 or OH.
In other embodiments, the lipid is a composition of one or more of the above lipids.
[00138] In certain embodiments, the lipid is prepared by reacting amine 32 with acrylate ND to form lipid ND32. In certain embodiments, the lipid ND32 is of one of the formulae below:

( i H2)11CH3 ( i H2)11CH3 0 --Tf HNI--, (CH2)11CH3 or HN OH

In other embodiments, the lipid is a composition of one or more of the above lipids.
[00139] In certain embodiments, the lipid is_ prepared by reacting amine 36 with acrylate ND to form lipid ND36. In certain embodiments, the lipid ND36 is of one of the formulae below:

( i H2)11CH3 ( i H2)11CH3 OH .
O ZZTf HNIN, HN
(CH2)11CH3 or OH.
In other embodiments, the lipid is a composition of one or more of the above lipids.
[00140] In certain embodiments, the lipid is prepared by reacting amine 98 with acrylate ND to form lipid ND98. In certain embodiments, the lipid ND98 is of one of the formulae below:

( i H2)11CH3 ( i H2)11CH3 N-'-~ N_-~ N--'~ N N/(CH2)11CH3 H
O --Yf (CH2)11CH3 0 i N H 0 i N H
(CH2)11CH3 (CH2)11L;H3 ( i H2)11CH3 N N _-~ N--'~ N N ,(CH2)11CH3 H
0 ~Yf HNI--, (CH2)11CH3 0 i NH 0 i N H
(CH2)11CH3 (CH2)11CH3 (CH2)11CH3 H2)11CH3 (CH2)11CH3 H2)11CH3 N N N--,~ N N /(CH2)11CH3 H H
O ~~f HN
(CH2)11CH3 0 i N H
(CH2)11CH3 ( i H2)11CH3 H
N-"~ NNNH

HN
~(CH2)11CH3 0 i N H 0 i N H
(CH2)11CH3 (CH2)11CH3 ( i H2)11CH3 H
/(CH2)11CH3 N N N N N
H H
0~Tf HN
(CH2)11CH3 0 i NH
(CH2)11CH3 ( i H2)11CH3 H
HN~~ N N "-~ N ,(CH2)110H3 N

0 iN H 0 'N H
(CH2)11CH3 ((;H2)11CH3 ( i H2)11CH3 H
NNN '11-~ N/(CH2)11CH3 H

O 'Yf HNN~-, (OH2)110H3 0 i N H

(CH2)11CH3 NNN "-~ N/(OH2)110H3 H

O -:Yf HNINI
(OH2)110H3 0 i N H
(OH2)110H3 (CH2)11CH3 H2)11CH3 H
HNN~~NH

I I
(CH2)11CH3 (CH2)11CH3 ( i H2)11CH3 (CH2)11CH3 H2)11CH3 N--'~H-~~N~~NH2 0 Z~Tf HN
\(CH2)11CH3 ( i H2)11CH3 H
N--'~ NNNH2 0 --ZTf (CH2)11CH3 0 i N H
(UH2)11CH3 ( CH2)11CH3 H2NN~NH2 I
t(;H2)11CH3 (CH2)11CH3 H2)11CH3 H
HNNN\~NH2 i (CH2)11CH3 (i H2)11CH3 H
N--'~ N--~ N\~N H2 HN
(CH2)11CH3 H H
N-"~ N---~ N--'~ N H
H

O -Z:~rf HNIN, (CH2)11CH3 0 i N H

(CH2)11 CH3 H H
N--'~HN N~~NH2 0 -Z~f HN~
(CH2)11CH3 or H

(UH2)11CH3 In other embodiments, the lipid is a composition of one or more of the above lipids.
[00141] In certain embodiments, the lipid is prepared by reacting amine 94 with acrylate ND to form lipid ND94. In certain embodiments, the lipid ND94 is of one of the formulae below:

( i H2)11CH3 ( i H2)11CH3 N
NH

HN
HN NH
(CH2)11CH3 or N~

In other embodiments, the lipid is a composition of one or more of the above lipids.
[00142] In certain embodiments, the lipid is prepared by reacting amine 95 with acrylate ND to form lipid ND95. In certain embodiments, the lipid ND95 is of one of the formulae below:
Me 0 NN "-~ N/(CH2)11CH3 H
O ':Yf HNI-I
(CH2)11CH3 0 i NH
(CH2)11CH3 Me I
N
'*'~ N H

0 -)--f HNI--, (CH2)11CH3 0 i NH

(CH2)11CH3 Me I
Me HN
NH
N

HN I
(CH2)11CH3 (CH2)11CH3 or Me 0 HN N N /(CH2)11CH3 H

I
(CH2)11CH3 In other embodiments, the lipid is a composition of one or more of the above lipids.
[00143] In certain embodiments, the lipid is prepared by reacting amine 96 with acrylate ND to form lipid ND96. In certain embodiments, the lipid ND96 is of one of the formulae below:

Me N "-~ /(CH2)11CH3 N N

I I
(CH2)11 CH3 (CH2)11 CH3 Me,-, N NH

I I
(CH2)11L;H3 (L;H2)11CH3 MeN., N NH2 H i NH
Me I I
(CH2)11CH3 (CH2)11CH3 or Me H N "-~ H /(CH2)11CH3 I
(CH2)11CH3 In other embodiments, the lipid is a composition of one or more of the above lipids.
[00144] In certain embodiments, the lipid is prepared by reacting amine 99 with acrylate ND to form lipid ND99. In certain embodiments, the lipid ND99 is of one of the formulae below:

( I H2)11CH3 N--"~ N N-,(CH2)11CH3 H

HN
~(CH2)11CH3 0 i N H
(CH2)11CH3 ( i H2)11CH3 N
N"~ NH

O ~rf HN~
(OH2)11CH3 0 i N H

(CH2)11CH3 ( i H2)11CH3 ( i H2)11CH3 HN 0 HN
NH
N

-~Tf 0 N H
H N ~ I .
(CH2)11CH3 (CH2)11CH3 or ( i H2)11CH3 HN"'~NH2, In other embodiments, the lipid is a composition of one or more of the above lipids.
In certain embodiments, ND99 is treated with Mel or another alkylating agent to form lipids of the formulae:

( I H2)11CH3 NMe+Ne+/(CH2)11CH3 N
H
0 -;~~f HN~
(CH2)11CH3 0 i NH
(OH2)110H3 ( i H2)11CH3 HN O
NMe/
NMe2+
0 -~'f HN

(CH2)11CH3 0 i NH
(CH2)11CH3 (CH2)11CH3 H2)11CH3 (CH2)11CH3 H2)11CH3 HN 0 NMe2/~
NMe2+
NMe/
NMe3+

HN I
(CH2)11CH3 (CH2)11CH3 or ( i H2)11CH3 HN O

+Me2 ~'~NMe3+

[00145] In certain embodiments, the lipid is prepared by reacting amine 100 with acrylate ND to form lipid ND 100. In certain embodiments, the lipid ND 100 is of one of the formulae below:

H N --~ N N N~(CH2)11CH3 I H
(CH2)11CH3 I I
(CH2)11CH3 (CH2)11CH3 H N J-"~ N N H
I
(CH2)11CH3 I I
((;H2)11CH3 (CH2)11CH3 H i N N NH2 (CH2)11CH3 I
(CH2)11CH3 0 iN H 0 iN H 0 iH
(L;H2)11CH3 (CH2)11CH3 or (CH2)11L;H3.
In other embodiments, the lipid is a composition of one or more of the above lipids.
[00146] In certain embodiments, the lipid is prepared by reacting amine 103 with acrylate ND to form lipid ND 103. In certain embodiments, the lipid ND 103 is of one of the formulae below:

( i H2)11CH3 H
HON~~OH HONNOH

I I
(CH2)11CH3 or ((;H2)11CH3 In other embodiments, the lipid is a composition of one or more of the above lipids.
[00147] In certain embodiments, the lipid is prepared by reacting amine 109 with acrylate ND to fomz lipid ND 109. In certain embodiments, the lipid ND 109 is of one of the formulae below:

( i H2)11CH3 HN 0 (CH2)11CH3 H2)11CH3 HN O
N N /~OH

H

_Yf HN1-.
(CH2)11CH3 0 NH
I HN
(CH2)11L;H3 (CH2)11CH3 H
N
N H
0 N"-"'-."N,-,,,/OH
H

)1f HN

(CH2)11CH3 0 NH
I HN
(CH2)11CH3 (CH2)11CH3 or H2N /,,,_/OH
N

I
(L; H2)11CH3.
In other embodiments, the lipid is a composition of one or more of the above lipids.
[00148] In certain embodiments, the lipid is prepared by reacting amine 98 with acrylate NE to form lipid NE98. In certain embodiments, the lipid NE98 is of one of the formulae below:

(CH2)12CH3 H2)12CH3 (CH2)12CH3 N--'~ N -,~N _,_~ N N/(CH2)12CH3 H
0~Yf HN~-, (CH2)12CH3 O j N H 0 N H
(CH2)12CH3 (CH2)12CH3 ( i H2)12CH3 H
N",/"-"'N~/N~~N N/(CH2)12CH3 H
O

HN
(CH2)12CH3 0 i N H O i N H
(CH2)12CH3 (CH2)12CH3 (CH2)12CH3 H2)12CH3 (CH2)12CH3 H2)12CH3 N~./~~ N~~ N N ,-(C H2)12CH3 H H
0 -'-If HN
(CH2)12CH3 0 ' N H
(CH2)12CH3 ( i H2)12CH3 H
NNN--'~ N H
O --Tf HN

(CH2)12CH3 0 i N H 0 i N H
(CH2)12CH3 (CH2)12CH3 (CH2)12CH3 H2)12CH3 H
/(CH2)12CH3 N N N N N
H H

HN
(CH2)12CH3 0 i H
(CH2)12CH3 ( i H2)12CH3 H
/(CH2)12CH3 HN N N ll-~ N
H

I I
(CH2)12CH3 (CH2)12CH3 (CH2)12CH3 H2)12CH3 H
NNNN N/(CH2)12CH3 H H

HNI--, (CH2)12CH3 0 i N H
(CH2)12CH3 H H
N ""'N /(CH2)12CH3 ~N ~~N N
H H
O -~~f HN
N", (CH2)12CH3 0 i N H
(CH2)12CH3 ( i H2)12CH3 H
HNNNNH

I I
(CH2)12CH3 (CH2)12CH3 (CH2)12CH3 H2)12CH3 (CH2)12CH3 H2)12CH3 N--'~H~~NNH2 HNI~-I(CH2)12CH3 (i H2)12CH3 H
NN--~NH2 0 )If HNNl~l (CH2)12CH3 0 i N H
(CH2)12CH3 (CH2)12CH3 HN' 0 I
(CH2)12CH3 ( i H2)12CHa H
HNN~~NH2 I
(CH2)12CH3 (CH2)12CH3 H2)12CH3 N-"~ N_~ N--'~NH2 H

O -~If HN
\ (CH2)12CH3 H H
N--'~ NN--'~NH
H

0 )-f HN i H
~(CH2)12CH3 0 N (CH2)12CH3 H H
N-'~H~~N-~~NH2 0 '~Zyf HNI-I(CH2)12CH3 or H
H2NNN-~'~NH2 O NH
(CH2)12CH3 In other embodiments, the lipid is a composition of one or more of the above lipids.
[00149] In certain embodiments, the lipid is prepared by reacting amine 94 with acrylate NE to forin lipid NE94. In certain embodiments, the lipid NE94 is of one of the formulae below:
(CH2)12CH3 H2)12CH3 (CH2)12CH3 H2)12CH3 N /
NH
0 N, HN
/
HN NH
(CH2)12CH3 or N-,,/ .

In other embodiments, the lipid is a composition of one or more of the above lipids.
[00150] In certain embodiments, the lipid is prepared by reacting amine 95 with acrylate NE to form lipid NE95. In certain embodiments, the lipid NE95 is of one of the formulae below:
Me 0 N --'~ N N / (CH2)12CH3 H
0 -.Tf HN
(CH2)12CH3 0 i H
(CH2)12CH3 Me I
N
NH

HNN., (CH2)12CH3 0 i H
(CH2)12CH3 Me I
Me HN
I NH
N

O ZZ:Yf HN ~
~(CH2)12CH3 (CH2)12CH3 or Me 0 HN N N /(CH2)12CH3 H
0. NH
I
(CH2)12CH3 In other embodiments, the lipid is a composition of one or more of the above lipids.
[00151] In certain embodiments, the lipid is prepared by reacting amine 96 with acrylate NE to form lipid NE96. In certain embodiments, the lipid NE96 is of one of the formulae below:

Me", N N N/(CH2)12CH3 H

I I
(CH2)12CH3 (CH2)12CH3 Me'.. N NH

0 ~H 0 NIH
(CH2)12CH3 (CH2)12CH3 Me,-. N NH2 H i NH

Me 0 ~ H 0 NH
(CH2)12CH3 (CH2)12L;H3 or Me,-, H N "-~ H ,(CH2)12CH3 I
(CHz)12CH3 In other embodiments, the lipid is a composition of one or more of the above lipids.
[00152] In certain embodiments, the lipid is prepared by reacting amine 99 with acrylate NE to form lipid NE99. In certain embodiments, the lipid NE99 is of one of the formulae below:
(CH2)12CH3 H2)12CH3 N N N ,,(CH2)12CH3 H
0 --Tf HN
(CH2)12CH3 0 ~ H
(CH2)12CH3 (CH2)12CH3 H2)12CH3 HN O
N
NH

O IzYf HNI--, (CH2)12CH3 0 i N H

(CH2)12CH3 (CH2)12CH3 ( i H2)12CH3 HNl 0 HN
NH
N
~~~NH2 _Yf 0 NH
HNI--, I
(CH2)12CH3 (CH2)12CH3 or (CH2)12CH3 H2)12CH3 HN"-""~"NH2, In other embodiments, the lipid is a composition of one or more of the above lipids.
In certain embodiments, NE99 is treated with Mel or another alkylating agent to forni lipids of the formulae:

(CH2)12CH3 H2)12CH3 O
MeNe+~N/(CH2)12CH3 H
O

HN
I--, (CH2)12CH3 0 i N H
(CH2)12CH3 ( i H2)12CH3 NMe/
NMe2+
0 _Yf HN
(CH2)12CH3 0 'N H
(CH2)12CH3 ( i H2)12CH3 (CH2)12CH3 H2)12CH3 HN 0 NMe~NMe2+
Ne/
NMe3+

HNINI(CH2)12CH3 (CH2)12CH3 or ( i H2)12CH3 +Me2N -'~NMe3+

[00153] In certain embodiments, the lipid is prepared by reacting amine 100 with acrylate NE to form lipid NE100. In certain embodiments, the lipid NE100 is of one of the formulae below:

HN N N (CH2)12CH3 H
(CH2)12CH3 I I
(CH2)12CH3 (CH2)12CH3 H i N N N H H i N NH2 (CH2)12CH3 (CH2)12L;H3 I I I
(CH2)12CH3 (CH2)12CH3 (CH2)12CH3 I I I
((;H2)12CH3 (CH2)12CH3 or (CH2)12CH3.
In other embodiments, the lipid is a composition of one or more of the above lipids.
[00154] In certain embodiments, the lipid is prepared by reacting amine 103 with acrylate NE to form lipid NE 103. In certain embodiments, the lipid NE103 is of one of the formulae below:

(CH2)12CH3 H2)12CH3 H
HOOH HO"-"' NN-~OH

I i (L;H2)12CH3 or (CH2)12CH3 In other embodiments, the lipid is a composition of one or more of the above lipids.
[00155] In certain embodiments, the lipid is prepared by reacting amine 109 witll acrylate NE to form lipid NE109. In certain embodiments, the lipid NE109 is of one of the formulae below:

(CH2)12CH3 H2)12CH3 HN 0 (CH2)12CH3 H2)12CH3 NN ,~,,"/OH

H
O
HN~
(CH2)12CH3 0 NH
-Tf i HN
(CH2)12CH3 ~(CH2)12CH3 H
N ~~/OH
N H

H

-~Tf HN
(CH2)12CH3 0 NH

I (CH2)12CHs HN~(CH2)12CHs or H2N,-,,,/OH

i (CH2)12CH3, In other embodiments, the lipid is a composition of one or more of the above lipids.
[00156] In certain embodiments, the lipid is prepared by reacting amine 1 with acrylate NF to form lipid NF 1. In certain embodiments, the lipid NF 1 is of one of the forinulae below:

(i H2)13CH3 (CH2)13CH3 H2)13CH3 N OMe H N 0 0 ~-f HN.~(CH2)1aCH3 or HN OMe In other embodiments, the lipid is a composition of one or more of the above lipids.
[00157] In certain embodiments, the lipid is prepared by reacting amine 10 with acrylate NF to form lipid NF 10. In certain embodiments, the lipid NF 10 is of one of the formulae below:

(CH2)13CH3 H2)13CH3 (CH2)13CH3 H2)13CH3 N
---~OMe HN 0 HN~
(CH2)13CH3 or HN~--~OMe, In other embodiments, the lipid is a composition of one or more of the above lipids.
[00158] In certain embodiments, the lipid is prepared by reacting amine 11 with acrylate NF to form lipid NF 11. In certain embodiments, the lipid NF10 is of one of the formulae below:

( i H2)13CH3 ( i H2)13CH3 ""~O Et 0 -~-f (CH2)13CH3 or '~~OEt.

In other embodiments, the lipid is a composition of one or more of the above lipids.
[00159] In certain embodiments, the lipid is prepared by reacting amine 20 with acrylate NF to form lipid NF20. In certain embodiments, the lipid NF20 is of one of the formulae below:

(CH2)13CH3 H2)13CH3 ( i H2)13CH3 HNIN, (CH2)13CH3 or HN OH

In other embodiments, the lipid is a composition of one or more of the above lipids.
[00160] In certain embodiments, the lipid is prepared by reacting amine 25 with acrylate NF to form lipid NF25. In certain embodiments, the lipid NF25 is of one of the formulae below:

( i H2)13CH3 (CH2)13CH3 H2)13CH3 OH

N
OH

OH
HNNI-I
(CH2)13CH3 or OH.

In other embodiments, the lipid is a composition of one or more of the above lipids.
[00161] In certain embodiments, the lipid is prepared by reacting amine 28 with acrylate NF to form lipid NF28. In certain embodiments, the lipid NF28 is of one of the formulae below:

( i H2~13CH3 (CH2)13CH3 H2)13CH3 OH

0 Zzllf HN HN
\(OH2?130H3 or OH.
In other embodiments, the lipid is a composition of one or more of the above lipids.
[00162] In certain embodiments, the lipid is prepared by reacting amine 32 with acrylate NF to form lipid NF32. In certain embodiments, the lipid NF32 is of one of the formulae below:

(CH2)13CH3 I

(CH2)13CH3 H2)13CH3 HN~(CH2)13CH3 or HN OH
In other embodiments, the lipid is a composition of one or more of the above lipids.
[00163] In certain embodiments, the lipid is prepared by reacting amine 36 with acrylate NF to form lipid NF36. In certain embodiments, the lipid NF36 is of one of the formulae below:
(CH2)13CH3 H2)13CH3 (CH2)13CH3 H2)13CH3 OH
O -~-f HN
~(CH2)13CH3 or HN OH.
In other embodiments, the lipid is a composition of one or more of the above lipids.
[00164] In certain embodiments, the lipid is prepared by reacting amine 60 with acrylate NF to form lipid NF60. In certain embodiments, the lipid NF60 is of one of the forinulae below:

( i H2)13CH3 Me~NN,Me ( i H2)130H3 0 i H N ,Me N
(CH2)13CH3 or M H

In other embodiments, the lipid is a composition of one or more of the above lipids.
[00165] In certain embodiments, the lipid is prepared by reacting amine 61 with acrylate NF to form lipid NF61. In certain embodiments, the lipid NF61 is of one of the formulae below:

( i H2)13CH3 EtNNEt ( i H2)13CH3 0 i N H EtNEt (CH2)13CH3 or H

In other embodiments, the lipid is a composition of one or more of the above lipids.
[00166] In certain embodiments, the lipid is prepared by reacting amine 63 with acrylate NF to form lipid NF63. In certain embodiments, the lipid NF63 is of one of the formulae below:

/(CH2)13CH3 N N "~~ N -11~~ N H1 I I H

(CH2)13CH3 Me Me or HN N NH
I I
(CH2)13CH3 Me Me.

In other embodiments, the lipid is a composition of one or more of the above lipids.
[00167] In certain embodiments, the lipid is prepared by reacting amine 64 with acrylate NF to form lipid NF64. In certain embodiments, the lipid NF64 is of one of the formulae below:

(CH2)13CH3 HN N N N
H
(CH2)13CH3 Et Et or HN N NH
I I
(CH2)13CH3 Et Et In other embodiments, the lipid is a composition of one or more of the above lipids.
[00168] In certain embodiments, the lipid is prepared by reacting amine 61 with acrylate NF to fonn lipid NF70. In certain embodiments, the lipid NF70 is of one of the formulae below:

H H
H3C(H2C)13 N N~~N N~(CH2)13CH3 0 0 or H
H3C(H2C)13 N NNH

In other embodiments, the lipid is a composition of one or more of the above lipids.
[00169] In certain embodiments, the lipid is prepared by reacting amine 86 with acrylate NF to form lipid NF86. In certain embodiments, the lipid NF86 is of one of the formulae below:

(CH2)13CH3 H2)13CH3 H2)13CH3 HN O (CH2)13CH3 HN O
N /~/OH
N HN ,-,,/0H
0 -)-f HN OH
(CH2)13CH3 or OH
In other embodiments, the lipid is a composition of one or more of the above lipids.
[00170] In certain embodiments, the lipid is prepared by reacting amine 87 with acrylate NF to form lipid NF87. In certain embodiments, the lipid NF87 is of one of the formulae below:

( i H2)13CH3 OH

(CH2)13CH3 H2)13CH3 N N~~OH H N 0 OH
O 'zYf HN~ HN N
(CH2)13CH3 or In other embodiments, the lipid is a composition of one or more of the above lipids.
[00171] In certain embodiments, the lipid is prepared by reacting amine 91 with acrylate NF to form lipid NF91. In certain embodiments, the lipid NF91 is of one of the formulae below:

( i H2)13CH3 0 (CH2)13CH3 O --Yf 0 HN1-1(CH2)13CH3 or HN NI

In other embodiments, the lipid is a composition of one or more of the above lipids.
[00172] In certain embodiments, the lipid is prepared by reacting amine 95 with acrylate NF to form lipid NF95. In certain embodiments, the lipid NF95 is of one of the formulae below:

( j H2)13CH3 N,,Me N
O ~~rf HNNl~' (CH2)13CH3 0 i N H
(CH2)13CH3 (CH2)13CH3 H2)13CH3 H
N"'~ N ,Me Me H
0 'f -~~Yf HN
~(CH2)13CH3 0 i N H
HN ~
(CH2)13CH3 (CH2)13CH3 ( i H2)13CH3 H2N~~~ Me HN,Me 0 i H

H or (UH2)13CH3.
In other embodiments, the lipid is a composition of one or more of the above lipids.
[00173] In certain embodiments, the lipid is prepared by reacting amine 96 with acrylate NF to form lipid NF96. In certain embodiments, the lipid NF96 is of one of the formulae below:

H N N N 'Me ( (CH2)13CH3 I i (CH2)13CH3 (CH2)13CH3 HN Jl"~ N N Me HN N ' Me I H
(CH2)13CH3 I I I
(CH2)13CH3 (CH2)13CH3 (CH2)13CH3 O

Me Me H i N H H2N N
(CH2)13CH3 I I
(CH2)13CH3 or (CH2)13CH3.

In other embodiments, the lipid is a composition of one or more of the above lipids.
[00174] In certain embodiments, the lipid is prepared by reacting amine 98 with acrylate NF to form lipid NF98. In certain embodiments, the lipid NF98 is of one of the formulae below:

(CH2)13CH3 H2)13CH3 (CH2)13CH3 N--'~ N_,-~ N--,~ N -"-~ N,-(CH2)13CH3 H
O ~-f HN

N H
(CH2)13CH3 0 I N H 0 I

(CH2)13CH3 (CH2)13CH3 (CH2)13CH3 H2)13CH3 H
/(CH2)13CH3 NN N N 11-~ N
H
O :Yf (CH2)13CH3 0 i N H 0 i N H
(CH2)13CH3 (CH2)13CH3 (CH2)13CH3 H2)13CH3 ( i H2)13CH3 NNN--~ N "-~ N,(CH2)13CH3 H H
0 ~)If HN
~(CH2)13CH3 0 i N H
(CH2)13CH3 ( i H2)13CH3 H
N--'~ N-~ N-"~ N H
0 ':Yf N H
(CH2)13CH3 0 I N H 0 I

(CH2)13CH3 (CH2)13CH3 (CH2)13CH3 H2)13CH3 H
NNNN N~(C H2)13CH3 H H
O 'Yf (OH2)13CH3 0 NH
I
(CH2)13CH3 ( i H2)13CH3 H
HNNNN "--~ N/(CH2)13CH3 H
0 NH 0 iN H

(CH2)13L;H3 (CH2)13CH3 ( i H2)13CH3 H
NNNN N/(C H2)13C H3 H H
O

HN
1~11 (CH2)130H3 0 i N H
(UH2)13UH3 H H
N,,~H H
NN "-~A N /(CH2)13CH3 0 -Z~Tf HN
INI
(GH2)130H3 0 i N H
(CH2)13CH3 ( i H2)13CH3 H
HNNN"-"""NH

I I
(CH2)13CH3 (CH2)13CH3 (CH2)13CH3 H2)13CH3 ( i H2~13CH3 N--'~HNNH2 O

HN
I-N (CH2)130H3 ( i H2)13CH3 H
N \~ N --'.~NH2 O -~Zzyf HNINI
(CH2)13CH3 0 i N H
(CH2)13CH3 ( i H2)13CH3 HZN--'~NH2 I
(CH2)13CH3 ( i H2)13CH3 HN O

H
HNN., ~ NH2 I
(CH2)13CH3 ( i H2)13CH3 HN O
H
NNN'~~NH2 H

HNNII(CH2)13CH3 NNNH
H

O ~Tf HNN~l (GH2)130H3 0 i N H
(CH2)130N3 NN'-~ NN"-~NH2 H
0 _Yf HN
~(CH2)13CH3 or H
HZNN~~NH2 {CH2113C~3 In other embodiments, the lipid is a composition of one or more of the above lipids.
1001751 In certain embodiments, the lipid is prepared by reacting amine 99 with acrylate NF to form lipid NF99. In certain embodiments, the lipid NF99 is of one of the formulae below:

( i H2)13CH3 N.~~N N,(GHz)13CH3 H
0 ~~rf HN
'-1 (CH2)13CH3 0 ' N H
tCt12)13C"3 (~ H2)13CH3 HN

N
N H
O -~~f HN
(CH2)13CH3 0 i N H
(CH2)13CH3 (CH2)13CH3 (CH2)13CH3 H2)13CH3 HN' 0 HN
NH
N
"~~ NH2 HNIII I
(CH2)13CH3 (CH2)13CH3 or (CH2)13CH3 H2)13CH3 HN
NH2, In other embodiments, the lipid is a composition of one or more of the above lipids.
In certain embodiments, NF99 is treated with Mel or another alkylating agent to form lipids of the formula:

( i H2)13CH3 NMe/Me+.~~ /(CH2)13CH3 N N
H
O Z:Yf HNNl-~
(CH2)130H3 0 i N H
(CH2)13CH3 ( i H2)13CH3 NMe/
NMe2+

HN

(CH2)13CH3 0 i N H
(CH2)13CH3 ( i H2)13CH3 (CH2)13CH3 H2)13CH3 HN 0 HN O

Me2/~
NMe2+
NMe NMe3+

HNN~' I
(CH2)13CH3 (CH2)13CH3 or (CH2)13CH3 H2)13CH3 Me2/~
NMe3+
[00176] In certain embodiments, the lipid is prepared by reacting amine 100 with acrylate NF to form lipid NF 100. In certain embodiments, the lipid NF 100 is of one of the formulae below:

HN N N N~(CH2)13CH3 I H
(UH2)13CH3 I I
~UH2)13UH3 (UH2)13CH3 H N )L-'~ N N H
I
(CH2)13CH3 I I
(CH2)13CH3 (L; H2)13CH3 O

H i N N NH2 H N N H
(UH2)13CH3 I I I
(CH2)13CH3 (CH2)13CH3 (CHO13CH3 or ( (CH2)13CH3.

In other embodiinents, the lipid is a composition of one or more of the above lipids.
[00177] In certain embodiments, the lipid is prepared by reacting amine 103 with acrylate NF to forni lipid NF103. In certain embodiments, the lipid NE 103 is of one of the formulae below:

(CH2)13CH3 H2)13CH3 H
H0N-"~0H HO-~NN"-"'~~OH

I I
(CH2)13CH3 or (CH2)13CH3 In other embodiments, the lipid is a composition of one or more of the above lipids.
[00178] In certain embodiments, the lipid is prepared by reacting amine 109 with acrylate NF to form lipid NF 109. In certain embodiments, the lipid NF 109 is of one of the formulae below:

(CH2)13CH3 H2)13CH3 HN 0 (CH2)13CH3 H2)13CH3 N
N ~/0H
0 fNoH
H

~~Yf HNN~' (CH2)13CH3 0 NH
-~Tf I HN
(OH2)13CH3 ~(CH2)13CH3 H
N /~/OH
N H

H

-)lf HN
I--, (CH2)13CH3 0 NH
I HN'-1 (CH2)13CH3 (CH2)13CH3 or H2N N /-,,/OH

I
(CH2)13CH3.
In other embodiments, the lipid is a composition of one or more of the above lipids.
[00179] In certain embodiments, the lipid is prepared by reacting amine 61 with acrylate NG to form lipid NG61. In certain embodiments, the lipid NG61 is of one of the formulae below:
(CH2)14CH3 H2)14CH3 H2)14CH3 N~~~ Et (CH2)14CH3 Et N HN 0 0 NH N Et I
(CH2)14CH3 or Et H
In other embodiments, the lipid is a composition of one or more of the above lipids.
[00180] In certain embodiments, the lipid is prepared by reacting amine 64 with acrylate NG to form lipid NG64. In certain embodiments, the lipid NG64 is of one of the formulae below:

HN )L"~ N N "~~ N,,(CH2)14CH3 I I I H
(CH2)14CH3 Et Et or HN N NH
I ( I
(CH2)14CH3 Et Et In other embodiments, the lipid is a composition of one or more of the above lipids.

[00181] In certain embodiments, the lipid is prepared by reacting amine 77 with acrylate NG to form lipid NG77. In certain embodiments, the lipid NG77 is of one of the formulae below:

(CH2)14CH3 HN O

(CH2)14CH3 HN O
N

O MeN
HN
HN
(CH2)14CH3 or MeN

In other embodiments, the lipid is a composition of one or more of the above lipids.
[00182] In certain embodiments, the lipid is prepared by reacting amine 86 with acrylate NG to form lipid NG86. In certain embodiments, the lipid NG86 is of one of the formulae below:

(i H2)14CH3 H2)14CH3 HN O (CH2)14CH3 HN O
N OH
N HN~~ OH

HNI-I OH
(CHO14CH3 or OH
In other embodiments, the lipid is a composition of one or more of the above lipids.
[00183] In certain embodiments, the lipid is prepared by reacting amine 87 with acrylate NG to form lipid NG87. In certain embodiments, the lipid NG87 is of one of the formulae below:

( i H2)14CH3 HN O
OH

(CH2)14CH3 N N ~~ HN 0 OH OH
O

HN1-~I(OH2)14CH3 or HN N"-"~'~OH.
In other embodiments, the lipid is a composition of one or more of the above lipids.
[00184] In certain embodiments, the lipid is prepared by reacting amine 95 with acrylate NG to form lipid NG95. In certain embodiments, the lipid NG95 is of one of the formulae below:

( i H2)14CH3 N N ,Me HN~
(CH2)14CH3 0 i N H
(CH2)14CH3 (CH2)14CH3 H2)14CH3 H
N"-~ N ,Me N~~ Me O

H
HN
N H
HN,~ \(CH2)140H3 0 I
(CH2)14CH3 (CH2)14CH3 (CH2)14CH3 Hz)14CHs H2N~Me HN,Me 0 iH

H or kCH2)14CH3.
In other embodiments, the lipid is a composition of one or more of the above lipids.
[001851 In certain embodiments, the lipid is prepared by reacting amine 100 with acrylate NG to form lipid NG100. In certain embodiments, the lipid NG100 is of one of the formulae below:

HN )t"~~ N N "~~ N~(CH2)14CH3 I H
~CH2)14CH3 I ( (CH2)14CH3 (CH2)14CH3 H i N N N H H i N 'jl"~ N NH2 (CH2)14CH3 (CH2)14GH3 0 iN H 0 iN H 0 iN H
(CH2)14CH3 (CH2)14CH3 (CH2)14CH3 0 iN H 0 iN H 0 iN H
~CH2)14CH3 ~CH2)14CH3 or (CH2)14CH3.
In other embodiments, the lipid is a composition of one or more of the above lipids.
In certain embodiments, NG100 is alkylated with methyl iodide or another alkylating agent.

[00186] In certain embodiments, the lipid is prepared by reacting amine 62 with acrylate NP to form lipid NP62. In certain embodiments, the lipid NP62 is of one of the formulae below:
(CH2)15CH3 H2)15CH3 ~, /~ /iPr ( i H2)15CH3 iPr \/ ~N HN 0 NH /N~~ iPr O I
(CH2)15CH3 or iPr H

In other embodiments, the lipid is a composition of one or more of the above lipids.
[00187] In certain embodiments, the lipid is prepared by reacting amine 63 with acrylate NP to form lipid NP63. In certain embodiments, the lipid NP63 is of one of the formulae below:

HN N N N /(CH2)15CH3 I I I H
(CH2)15CH3 Me Me or HN N NH
I I I
(CHZ)15CH3 Me Me.

In other embodiments, the lipid is a composition of one or more of the above lipids.
[00188] In certain embodiments, the lipid is prepared by reacting amine 86 with acrylate NP to form lipid NP86. In certain embodiments, the lipid NP86 is of one of the formulae below:

(CH2)15CH3 H2)15CH3 (CH2)15CH3 H2)15CH3 HN O

N ,-, /OH
N HN~~

HN~ OH
(CH2)15CH3 or OH
In other embodiments, the lipid is a composition of one or more of the above lipids.
[00189] In certain embodiments, the lipid is prepared by reacting amine 87 with acrylate NP to form lipid NP87. In certain embodiments, the lipid NP87 is of one of the formulae below:

( ~ H2)15CH3 OH

( i H2)15CH3 N N H N O
OH OH

HN HN N~~~
~(CH2)15CH3 or ON.
In other embodiments, the lipid is a composition of one or more of the above lipids.
[00190] In certain embodiments, the lipid is prepared by reacting amine 96 with acrylate NP to form lipid NP96. In certain embodiments, the lipid NP96 is of one of the formulae below:

Hj N N N' Me ((;H2)15CH3 I I
(CH2)15CH3 (CH2)15CH3 HN N N__Me HN N__Me I H
(CH2)15CH3 I I I
(CH2)15CH3 (CH2)15CH3 (CH2)15CH3 H ~Me H2N N ~Me i H
N
(CH2)15CH3 I I
(CH2)15CH3 or (L;H2)15CH3.
In other embodiments, the lipid is a composition of one or more of the above lipids.
[00191] In certain embodiments, the lipid is prepared by reacting amine 98 with acrylate NP to form lipid NP98. In certain embodiments, the lipid NP98 is of one of the formulae below:

(CH2)15CH3 H2)15CH3 (CH2)15CH3 H2)15CH3 NN_--~ NN N~(CH2)15CH3 H
O 'zYf HN1~1 (CH2)15CH3 0 i N H 0 i N H
(CH2)15L;H3 (UH2)15CH3 ( i H2)15CH3 H
NN-~ NN/(OH2)15CH3 H
O :~Tf N H
(OH2)15CH3 0 I N H 0 I

(CH2)15CH3 (CH2)15CH3 (CH2)15CH3 H2)15CH3 (CH2)15CH3 H2)15CH3 N"'~ NN"'~ N "--~ N/(CH2)150H3 H H
0 IZZYf HN

(CH2)15CH3 0 i N H
(CH2)15CH3 ( i H2)15CH3 H
N N__,~ NN H

HN
N H
(CH2)150H3 0 I N H 0 I

(CH2)15CH3 (CH2)15CH3 ( i H2)15CH3 ,,(CH2)15CH3 N N N N N
H H
O --Yf HNI--, (CH2)15CH3 O i N H
(CH2)15CH3 ( i H2)15CH3 HN~~ N N "-~ /(CH2)15CH3 H

I i (CH2)15CH3 (CH2)15CH3 ( i H2)15CH3 0 H N~~ N~~~ (CH2)15CH3 H N H
O -:Yf HN

(CH2)15CH3 0 i NH
(CH2)15CH3 H H
NNNN N,(CH2)15CH3 H H

HN
(CH2)15CH3 0 i N H
(CH2)15CH3 (CH2)15CH3 H2)15CH3 H
HNNNNH

I I
(CH2)15CH3 (CH2)15CH3 (CH2)15CH3 H2)15CH3 (CH2)15CH3 H2)15CH3 NH~~NNH2 0 -.~Tf HN~
(CH2)15CH3 (CH2)15CH3 H2)15CH3 HN O

H
N--'~ NN--~~NH2 0 -)-f HNNI-I
(CH2)15CH3 0 i N H
(CH2)15CH3 ( i H2)15CH3 N~~ NH2 I
(CH2)15L;H3 (CH2)15CH3 H2)15CH3 H

I
(UH2)15UH3 (CH2)15CH3 H2)15CH3 HN O
H

0 '~~f HN
INI(CH2)15CH3 H H
NN-'-~NH
H

HN
"I
(CH2)15CH3 0 i NH
(CH2)15CH3 H H
N~~NH2 H

0 'zYf HN
(CH2)15CH3 or H

--'~NH2 (CH2)15CH3 In other embodiments, the lipid is a composition of one or more of the above lipids.
[00192] In certain embodiments, the lipid is prepared by reacting amine 99 with acrylate NP to form lipid NP99. In certain embodiments, the lipid NP99 is of one of the formulae below:

(CH2)15CH3 H2)15CH3 NN N/(OH2)150H3 H
0 )lf HN

(CH2)150H3 0 i NH
(CH2)15CH3 ( i H2)15CH3 N
0~Yf HNN~l (OH2)150H3 0 i N H
(CH2)15CH3 (iH2)15CH3 (CH2)15CH3 H2)15CH3 HN 0 HN
NH
N
"~~NH2 HN i (CH2)15CH3 (CH2)15CH3 or ( i H2)15CH3 HN "-"'~'NH2, In other embodiments, the lipid is a composition of one or more of the above lipids. In certain embodiments, NF99 is treated with MeI or another alkylating agent to form lipids of the formula:

( i H2)15CH3 O
HNM M/(CH2)15CH3 N N
H
O --Zllf HN
~(CH2)15CH3 0 i NH
(CH2)15L;H3 (CH2)15CH3 H2)15CH3 NMe/~
NMe2+
HN
(CH2)15CH3 0 i NH
(CH2)15CH3 ( i H2)15CH3 (CH2)15CH3 H2)15CH3 HN 0 NMe2/~
NMe2+
NMe NMe3+

O NH
HNI-I I
(CH2)15CH3 (CH2)15CH3 or (CH2)15CH3 H2)15GH3 NMe2/~~
NMe3+
[00193] In certain embodiments, the lipid is prepared by reacting amine 100 with acrylate NP to form lipid NP 100. In certain embodiments, the lipid NP 100 is of one of the formulae below:

HN N N (CH2)15CH3 H
(CH2)15CH3 I I
(CH2)15CH3 (CH2)15CH3 H i N )L~~ N N H HN) N NH2 (CH2)15CH3 (CH2)15CH3 I I I
(CH2)15CH3 (CH2)15CH3 (CH2)15CH3 (CH2)15CH3 (CH2)15CH3 or (CH2)15CH3.
In other embodiments, the lipid is a composition of one or more of the above lipids.
[00194] In certain embodiments, the lipid is prepared by reacting amine 103 with acrylate NP to form lipid NP 103. In certain embodiments, the lipid NP 103 is of one of the formulae below:

(CH2)15CH3 H
H 0 N/~/ N~~O H H O~\ NN0 H

I I
(CH2)15CH3 or (CH2)15CH3 In other embodiments, the lipid is a composition of one or more of the above lipids.
[00195] In certain embodiments, the lipid is prepared by reacting amine 31 with acrylate LD to form lipid LD3 1. In certain embodiments, the lipid LD31 is of one of the formulae below:

( i H2)11CH3 ( i H2)11CH3 0 'Yf 0 HN,~ ~ /OH
~(CH2)11CH3 or v 'O/ v In other embodiments, the lipid is a composition of one or more of the above lipids.
[00196] In certain embodiments, the lipid is prepared by reacting amine 98 with acrylate LD to form lipid LD98. In certain embodiments, the lipid LD98 is of one of the formulae below:

( i H2)11CH3 ( i H2)11CH3 O

/~~ /(CH2)11CH3 N N N N O

(CH2)11CH3 0 i 0 0 0 (CH2)11 CH3 (CH2)11 CH3 ( i H2)11CH3 N--'~ N N-'-~ N O /(CH2)11CH3 O

~(CH2)11CH3 0 i 0 0 i 0 ((;H2)11CH3 (CH2)11CH3 ( i H2)11CH3 ( i H2)11cH3 NNNN 0/(CH2)11CH3 H

0 lz~f O

(CH2)11CH3 0 0 (CH2)11CH3 ( i H2)11CH3 H ' NNNH
O --Tf O
INI
(CH2)11CH3 0 0 0 0 I I
(CH2)11CH3 (CH2)11CH3 ( i H2)11CH3 H
NNN~~N (CH2)11CH3 H

O ~-f O

(CH2)11CH3 0 0 (CH2)11CH3 ( i H2)11CH3 O O

H
/(CH2)11CH3 HN N N N O

(CH2)11CH3 (CH2)11CH3 ( i H2)11CH3 H
/(CH2)11CH3 N,'-~ N N N 0 H
O -Z~f (CH2)11CH3 0 i (CH2)11CH3 N--'~ N NN 0 /(CH2)11CH3 H

0 ~~rf ~(CH2)11CH3 0 i (CH2)11CH3 ( i H2)11CH3 H
HN--~ N-~ N~/~NH

(CH2)11CH3 (CH2)11CH3 ( i H2)11CH3 (CH2)11CH3 H2)11CH3 N--"~ HNNH2 0 ~-f ll~l (CH2)11CH3 ( i H2)11CH3 H
N--'~ NN-'-~NH2 0 4zYf (CH2)11CH3 0 ~ H
(CH2)11CH3 ( i H2)11CH3 O O

N'~'/ \~~NH2 O O

kCH2)11CH3 ( i H2)11CH3 O O

H
HNN.~~NH2 O O

(OH2)11CH3 ( i H2)11CH3 O O
H
N'/~~' N*,~ NN H2 H

1-,(CH2)11CH3 H H
NNN N H
H

O -Z::Tf ~(CH2)11CH3 0 0 ((;H2)11CH3 H H
H
N N ---/\ NH2 0 --Zyf ~(CHz)11CH3 or H

((;H2)11CH3 . In other embodiments, the lipid is a composition of one or more of the above lipids.
[00197] In certain embodiments, the lipid is prepared by reacting amine 99 with acrylate LD to form lipid LD99. In certain embodiments, the lipid LD99 is of one of the formulae below:

( i H2)11CH3 NN ,,(C H2)11 C H3 O 4z:Yf ~(CH2)11CH3 0 i ((jH2)11 CH3 ( ~ H2)11CH3 N
~'~NH

O 'Yf O

(CH2)11CH3 0 0 (CH2)11CH3 (CH2)11CH3 H2)11CH3 ( ~ H2)11CH3 0 0 HN
NH

0 \(CH2)11CH3 (CH2)11CH3 or ( i H2)11CH3 NH2, In other embodiments, the lipid is a composition of one or more of the above lipids. In certain embodiments, LD99 is treated with Mel or another alkylating agent to form lipids (QD99) of the formula:

( H2)11CH3 Me/~Ne/(CH2)11CH3 (CH2)11CH3 0 0 (CH2)11CH3 (CH2)13CH3 H2)13CH3 Me\~NMe2+

0 -Z~f (CH2)13CH3 0 0 (CH2)13CH3 ( i H2)11CH3 ( H2)11CH3 0 0 N Me2/~
NMe2+
NMe/~
NMe3+

0 ~
~(CH2)11CH3 (CH2)11CH3 or (CH2)11CH3 H2)11CH3 NMe2/~
NMe3+, [00198] In certain embodiments, the lipid is prepared by reacting amine 100 with acrylate LD to form lipid LD 100. In certain embodiments, the lipid LD 100 is of one of the formulae below:

O J""~ N N 0/(CH2)11CH3 I
(CH2)11CH3 (CH2)11CH3 (CH2)11CH3 0 --~ i N N H i N NH2 (CH2)11CH3 (CH2)11CH3 (CH2)11CH3 (CH2)11CH3 (CH2)11CH3 (CH2)11CH3 (CH2)11CH3 or (CN2)11CH3. In other embodiments, the lipid is a composition of one or more of the above lipids. In certain embodiments, LD 100 is treated with Mel or another alkylating agent to form lipids (QD 100) of the formula:

N e+~~ N e+-~p/ (C H2)11 C H 3 0 J,'~ I

(CH2)11CH3 (CH2)11CH3 (CH2)11CH3 M e+~~
i N NMe2+
(CH2)11CH3 (CH2)11CH3 (CH2)11CH3 Me+'-~~
)L"~ N NMe3+ +Me2N NMe2+
i (CH2)11CH3 (CH2)11CH3 (L;H2)11CH3 (CH2)11CH3 or +Me3N NMe2+

(CH2)11CH3.

[00199] In certain embodiments, the lipid is prepared by reacting amine 87 with acrylate LE to form lipid LE87. In certain embodiments, the lipid LE87 is of one of the formulae below:

( i H2)12CH3 O
OH

(CH2)12CH3 H2)12CH3 N N """-'~'O O
OH OH
0 -::~Tf HN N
(CH2)12CH3 or In other embodiments, the lipid is a composition of one or more of the above lipids.
[00200] In certain embodiments, the lipid is prepared by reacting amine 94 with acrylate LE to form lipid LE94. In certain embodiments, the lipid LE94 is of one of the formulae below:

H2)12CH3 ( I
(CH2)12CH3 H2)12CH3 o O 0 0 N
NH HN

O 'zYf N ---/ N, 1~r NH

1-1 (CH2)12CH3 or N --:z .

In other embodiments, the lipid is a composition of one or more of the above lipids.
[00201] In certain embodiments, the lipid is prepared by reacting amine 31 with acrylate LF to form lipid LF3 1. In certain embodiments, the lipid LF31 is of one of the formulae below:

( ~ H2)13CH3 ( i H2)13CH3 N~~ /~/OH 0 O
O -Z~f 0 HN' ~ ~ 'OH
'(CH2)13CH3 or v ' / v In other embodiments, the lipid is a composition of one or more of the above lipids.
[00202] In certain embodiments, the lipis is prepared by reacting amine 94 with acrylate LF to form lipid LF94. In certain embodiments, the lipid LF94 is of one of the formulae below:

( i H2)13CH3 ( i H2)13CH3 N
N H
O N
HN
HN NH
(CH2)13CH3 or N-,-/ . In other embodiments, the lipid is a composition of one or more of the above lipids.
[00203] In certain embodiments, the lipid is prepared by reacting amine 95 with acrylate LF to form lipid LF95. In certain embodiments, the lipid LF95 is of one of the formulae below:

(CH2)13CH3 H2)13CH3 O O
N~~ Me O

O

(CH2)130H3 O i (CH2)13CH3 ( i H2~13CH3 H
NN Me 1N-"~~Me O

O (CH2)13CH3 0 i 0 ~(CH2)13CH3 (CH2)13CH3 (OH2)13 0 3 H2N~~ ~Me N
H"~~ ~Me O I
H or (CH2)13CH3. In other embodiments, the lipid is a composition of one or more of the above lipids.
[00204] In certain embodiments, the lipid is prepared by reacting amine 99 with acrylate LF to forin lipid LF 99. In certain embodiments, the lipid LF99 is of one of the formulae below:

( i H2)13CH3 N N "-~~ 0 /(CH2)13CH3 0 -~f (CH2)13CH3 0 0 (CH2)13CH3 (CH2)13CH3 H2)13CH3 N
"~~ NH

0 --ZTf O

(CH2)13CH3 0 0 (CH2)13CH3 ( i H2)13CH3 (CH2)13CH3 H2)13CH3 0 0 HN~~
NH

0 (CH2)13CH3 (CH2)13CH3 or ( i H2)13CH3 HN ~'~NH2, In other embodiments, the lipid is a composition of one or more of the above lipids. In certain embodiments, LF99 is treated with MeI or another alkylating agent to form lipid (QF99) of the formula:

( i H2)13CH3 HNM M/(CH2)13CH3 O

(CH2)13CH3 0 0 (CH2)13CH3 ( i H2)13CH3 Me/~
NMe2+
0 -Yf N., (CH2)13GH3 0 0 (UH2)13GH3 (CH2)13CH3 H2)13CH3 (CH2)13CH3 H2)13CH3 0 0 NMe2/~
NMe2+
NMe/
NMe3+
O zz~Tf 0 0 ONN(CH2)13CH3 (CH2)13CH3 or (CH2)13CH3 H2)13CH3 NMe2\;--~NMe3+

[00205] In certain embodiments, the lipid is prepared by reacting amine 32 with acrylate LG to form lipid LG32. In certain embodiments, the lipid LG32 is of one of the formulae below:
( i H2)14eH3 (i H2)14CH3 O ~~rf \(OH2)140H3 or HN 0H In other embodiments, the lipid is a composition of one or more of the above lipids.
[00206] In certain embodiments, the lipid is prepared by reacting amine 77 with acrylate LG to form lipid LG77. In certain embodiments, the lipid LG77 is of one of the formulae below:

(CH2)14CH3 H2)14CH3 O O

( i H2~14CH3 N

"'f HN
O
1-1(CH2)14CH3 or . In other embodiments, the lipid is a composition of one or more of the above lipids.
[00207] In certain embodiments, the lipid is prepared by reacting amine 80 with acrylate LG to form lipid LG80. In certain embodiments, the lipid LG80 is of one of the formulae below:

(CH2)14CH3 H2)14CH3 (CH2)14CH3 H2)14CH3 O -Z~Yf 0~ HN I
(CH2)14CH3 or In other embodiments, the lipid is a composition of one or more of the above lipids.
[00208] In certain embodiments, the lipid is prepared by reacting amine 96 with acrylate LG to form lipid LG96. In certain embodiments, the lipid NG96 is of one of the formulae below:

i N N~Me (CH2)14CH3 0 i 0 o ~

(CH2)14CH3 (CH2)14L;H3 N ~Me HN N ' Me i N

(CH2)14CH3 (CH2)14CH3 (CH2)14CH3 (CH2)14CH3 N Me H2N N ,Me i H ~

(CH2)14CH3 0 0 o 0 (CH2)14CH3 or (CH2)14CH3.

In other embodiments, the lipid is a composition of one or more of the above lipids.
[00209] In certain embodiments, the lipid is prepared by reacting amine 100 with acrylate LG to form lipid LG100. In certain embodiments, the lipid LG100 is of one of the formulae below:

0 N N 0 ,(CH2)140H3 {
(CH2)14CH3 I I
(CH2}14CH3 (CH2)14CH3 0 )L"~'~ i N N H i -"~ N NH2 (CH2)14CH3 (CH2)14CH3 (CH2)14CH3 (CH2)14CH3 (CH2)14CH3 (CH2)14CH3 (L;H2)14CH3 or (CH2)14CH3. In other embodiments, the lipid is a composition of one or more of the above lipids. In certain embodiments, LG100 is treated with Mel or another alkylating agent to form lipids (QG100) of the formula:

Me}Ne}~~ /(CH2)14CH3 O N O
(CH2)14CH3 (L;H2)14CH3 (CH2)14CH3 M e+"~~
O N NMe2+
(CH2)14CH3 I I
(CH2)14CH3 (CH2)14CH3 p N e+~~~NMe3+ +Me2N NMe2+
(CH2)14CH3 O i 0 i 0 i (GH2)14CH3 (CH2)14CH3 (CH2)14CH3 or +Me3N NMe2+

(CH2)14CH3.
[00210] In certain embodiments, the lipid is prepared by reacting amine 109 with acrylate LG to form lipid LG109. In certain embodiments, the lipid NG109 is of one of the formulae below:
( i H2)14CH3 O 0 (CH2)14CH3 H2)14CH3 N N ~,'/OH

H

~)'f \(CH2)14CH3 0 0 -Yf (CH2)14CH3 (CH2)14CH3 H
N~\ OH
N H
0 N N/~OH
H

'Yf O
\(CH2)14CH3 0 0 (CH2)14CH3 (CH2)14CH3 or H2N~~ ~/OH
N

(CH2)14CH3.
In other embodiments, the lipid is a composition of one or more of the above lipids.
[002111 In certain embodiments, the lipid is prepared by reacting amine 64 with acrylate LG to form lipid LG64. In certain embodiments, the lipid LG64 is of one of the formulae below:

0 )CH2)14CH3 N N
I I I
(UH2)14CH3 Et Et or O

O N NH
I I I
(CH2)14CH3 Et Et In other embodiments, the lipid is a composition of one or more of the above lipids.
[00212] In certain embodiments, the lipid is prepared by reacting amine 31 with acrylate LG to form lipid LG3 1. In certain embodiments, the lipid LG31 is of one of the formulae below:

( i H2)14CH3 ( i H2)14CH3 O

O HN OH
~(CH2)14CH3 or ~~0~~~ .
In other embodiments, the lipid is a composition of one or more of the above lipids.
[00213] In certain embodiments, the lipid is prepared by reacting ainine 32 with acrylate LG to form lipid LG32. In certain embodiments, the lipid NG32 is of one of the formulae below:

(i H2)14CH3 O

(CH2)14CH3 N OH O O
O

(OH2)14CH3 or HN OH
In other embodiments, the lipid is a composition of one or more of the above lipids.
Synthesis of Lipids [00214] The inventive lipids may be prepared by any method known in the art.
Preferably the lipids are prepared from commercially available starting materials, such acrylates or acrylamides, and amines. In another preferred embodiment, the lipids are prepared from easily and/or inexpensively prepared starting materials. As would be appreciated by one of skill in the art, the inventive lipids can be prepared by total synthesis starting from commercially available starting materials A
particular lipid may be the desired final product of the synthesis, or a mixture of lipids may be the desired final product.
[00215] In a particularly preferred embodiment, the inventive lipid is prepared via the conjugate addition of primary amines to acrylates or acrylamides. An exemplary reaction scheme is shown below:
O O
II R1 J), R1 2equiv. O + -{2~j~ ~
R

I

Any primary amine is useful in preparing inventive lipids. Primary amines useful in this invention include, but are not limited to, methylamine, ethylamine, isopropylamine, aniline, substituted anilines, and ethanolamine. The primary amine may be a bis(primary amine). Preferably, the amine is commercially available.
In certain embodiments, the amine used in the synthesis of the lipid is of the formula:

1 ~O,,,,NH2 24 OH HO,,~,NH2 77 ~~NH2 COY~ 25 HO:>--NH2 I\ NH2 NHz HO 79 O
7 0~ 26 HO,~6H 80 N,_,-~NH2 'O'-~NH2 28 HO~~NH2 81 N/--"NH2 11 o ~O NH2 NH2 31 HO'~O~~NH2 82 CN2 13 _0 HO"--~'NH2 HO--~N__-NH2 32 86 --l O-NH2 33 HO" '~' 'NH2 HO NH2 O I HO'--"N-r-l _0 87 HO 90 O N~~
HO'~~NH2 36 HO NH2 U NH2 21 NH 2 38 HO- 91 O~ ~,,NH2 ~-NH2 22 j~NH2 60 H NJ
N
H H 94 HN-'~NH2 ~
61 /--N~
H H 95 ~NNHZ
H
62 H~=~N~ 96 \N---NHa H H H

98 H2N'---H---_,N'/-NH2 NN
64 H H 99 HaN,NH2 H
75 NtiNH2 103 HO~~N-~,,N'-"-"OH
/ H
H
76 CN-,_,NH2 109 H2N,-~N'/~OH

95 'IN..... INHZ 112 HZN/H~iNHZ

96 '-H~~NHZ 113 HZN~~Ni~NHz H
98 HxN/-H-~N,''NH, 114 HzN~~N~NH=
H
99 HzNi-,,~NHz NH2 100 HzN~-'~~NH= N, H NH=
109 HZN,~iN-/'OH 61"NH, NH, 116 H,N~~NJ( 110 ~ NH2 NH= 117 NHZ
ll1 HzN"N-~N~/~N/\/NH' H H
[00216] Acrylate or acrylamide monomers that are useful in the present invention include any acrylates and acrylamides In certain embodiments, the acrylates or acrylamides are acrylates or acrylamides of straight chain alkyl groups.
In certain embodiments, the acrylate or acrylamide is of the formula:

LA NA N
H

LB NB
H

H
LD ND Nx/
H

LE NE
H

LF NF
NF H
O

LG.2 O0 NP H

LH NH NH N
H
In other embodiments, the acrylate or acrylamide may include branched, substituted, or cyclic aliphatic or heteroaliphatic groups. In certain embodiments, the acrylate or acrylamide is substituted with C 1-C6 alkyl group, halogens, amino groups, hydroxyl groups, alkoxy groups, etc.
[00217] In certain embodiments, the reaction is performed neat without the use of a solvent. In other embodiments, a solvent is used for the reaction. Both or one of the monomers is dissolved in an organic solvent (e.g., THF, CH2C12, MeOH, EtOH, CHC13, hexanes, toluene, benzene, CC14, glyme, diethyl ether, etc.). The resulting solutions are combined, and the reaction mixture is heated to yield the desired lipid.
In a particularly preferred embodiment, the reaction mixture is heated to temperature ranging from 50-150 C. In another particularly preferred embodiment, the reaction mixture is heated to approximately 95 C. The reaction may also be catalyzed.
For example, the reaction may be catalyzed by the addition of an acid, base, or metal. The reagents may be allowed to react for fours, days, or weeks. Preferably, the reaction is allowed to proceed from overnight (e.g., 8-2 hours) to 7 days.
[00218] In another particularly preferred embodiment, the inventive lipids are prepared by the conjugate addition of a bis(anline) to an acrylate. The bis(amine) may be a bis(secondary amine) or a bis(primary amine). En exemplary reaction scheme using bis(amines) is shown below:
O
RI

O~~ H5 5 n equiv. J.~ /+ A --~ A
Rl~\% 5 5 N~i N\
\R4 Rq Rl O
In certain embodiments, the reaction is performed neat without a solvent. In other embodiments, the reaction is performed in a solvent. One or both of the monomers are dissolved in an organic solvent (e.g., THF, CH2C12, MeOH, EtOH, CHC13, hexanes, CC14, glyme, diethyl ether, etc.). Organic solvents are preferred due to the susceptibility of polyesters to hydrolysis. The resulting solutions are combined, and the reaction mixture is heated to yield the desired lipid. In a particularly preferred embodiment, the reaction mixture is maintained at a temperature ranging from C. In another particularly preferred embodiment, the reaction mixture is heated to approximately 95 C. The reaction may also be catalyzed. For example, the reaction may be catalyzed by the addition of an acid, base, or metal.
1002191 In yet another particularly preferred embodiment, the inventive lipids are prepared by the conjugate addition of a poly(amine) to an acrylate or acrylamide.
The poly(amine) may include primary, secondary, tertiary, or quaternary amines. In certain embodiments, the poly(amine) contains only primary and secondary amines.
An exemplary reaction scheme using poly(amines) is shown below:
O
R 3 R4 R 3 Ra /1f } HN A,-tN Av"-NH ---> N A%rvN A-"-N
Ri H
n equiv, Io % o Io Rl R, R2 In certain embodiments, the reaction is performed with an excess of acrylate or acrylamide to fully funcationlize all amino groups of the poly(amine). In other embodiments, the equivalents of acrylate are limiting; therefore, all amino groups of the poly(amine) are not functionalized. In certain embodiments, the reaction is performed neat without a solvent. In other embodiments, the reaction is performed in a solvent. One or both of the monomers are dissolved in an organic solvent (e.g., THF, CH2C12, MeOH, EtOH, CHC13, hexanes, CCI4, glyme, diethyl ether, etc.).
Organic solvents are preferred due to the susceptibility of polyesters to hydrolysis.
The resulting solutions are combined, and the reaction mixture is heated to yield the desired lipid. In a particularly preferred embodiment, the reaction mixture is maintained at a temperature ranging from 50-150 C. In another particularly preferred embodiment, the reaction mixture is heated to approximately 95 C. The reaction may also be catalyzed. For example, the reaction may be catalyzed by the addition of an acid, base, or metal.
[00220] The synthesized lipid may be purified by any technique known in the art including, but not limited to, precipitation, crystallization, chromatography, distillation, etc. In a particularly preferred embodiment, the lipid is purified through repeated precipitations in organic solvent (e.g., diethyl ether, hexane, etc.). In a particularly preferred embodiment, the lipid is isolated as a salt. The lipid is reacted with an acid (e.g., an organic acid or inorganic acid) to form the corresponding salt.
In certain embodiments, the tertiary amine is alkylated to form a quaternary ammonium salt of the lipid. The tertiary amines may be alkylated with any alkylating agent, for example, alkyl halides such as methyl iodide may be used to from the quaternary amino groups. The anion associated with the quaternary amine may be any organic or inorganic anion. Preferably, the anion is a pharmaceutically acceptable anion.
[00221] In certain embodiments, the reaction mixture results in a mixture of isomers with varying numbers and positions of acrylate tails. Such mixtures of products may be used as is, or a single isomer may be purified from the reaction mixture. When an amine is not exhaustively alkylated, the resulting primary, secondary, or tertiary amines may be further reacted witli another acrylate, acrylamide, or other electrophile. The resulting lipid may then be optionally purified.
[00222] In certain embodiments, a desired lipid is prepared by traditional total synthesis. In certain embodiments, a commercially available amine is the starting material. One or more amino groups of the amine are optionally protected. The unprotected amino groups are reacted with a acrylate or acrylamide. The product is optionally purified. Protecting groups are removed, and the free amino groups are optionally reacted with another acrylate, acrylamide, or other electrophile.
Such a sequence may be repeated depending on the desired complexity of the inventive product being prepared. The final product may then be optionally purified.
[00223] In one embodiment, a library of different lipids is prepared in parallel.
A different amine and/or acrylate is added to each vial in a set of vials or to each well of a multi-well plate used to prepare the library. The array of reaction mixtures is incubated at a temperature and length of time sufficient to allow formation of the lipids to occur. In one embodiment, the vials are incubated at approximately overnight. In other embodiments, the vials are incubated from 1 to 7 days at approximately 95 C. The lipids may then be isolated and purified using techniques known in the art. The lipids may then be screened using high-throughput techniques to identify lipids with a desired characteristic (e.g., solubility in water, solubility at different pH, ability to bind polynucleotides, ability to bind heparin, ability to bind small molecules, ability to form microparticles, ability to increase tranfection efficiency, etc.). In certain embodiments the lipids may be screened for properties or characteristics useful in gene therapy (e.g., ability to bind polynucleotides, increase in transfection efficiency).

Polynucleotide Coniplexes [00224] The ability of cationic compounds to interact with negatively charged polynucleotides through electrostatic interactions is well known. Cationic lipids such as Lipofectamine have been prepared and studied for their ability to complex and transfect polynucleotides. The interaction of the lipid with the polynucleotide is thought to at least partially prevent the degradation of the polynucleotide.
By neutralizing the charge on the backbone of the polynucleotide, the neutral or slightly-positively-charged complex is also able to more easily pass through the hydrophobic membranes (e.g., cytoplasmic, lysosomal, endosomal, nuclear) of the cell. In a particularly preferred embodiment, the complex is slightly positively charged.
In another particularly preferred embodiment, the complex has a positive ~-potential, more preferably the C-potential is between +1 and +30.
[00225) The lipids of the present invention possess tertiary amines. Although these amines are hindered, they are available to interact with a polynucleotide (e.g., DNA, RNA, synthetic analogs of DNA and/or RNA, DNA/RNA hydrids, etc.).
Polynucleotides or derivatives thereof are contacted with the inventive lipids under conditions suitable to form polynucleotide/lipid complexes. The lipid is preferably at least partially protonated so as to form a complex with the negatively charged polynucleotide. In a preferred embodiment, the polynucleotide/lipid complexes form nanoparticles that are useful in the delivery of polynucleotides to cells. In certain embodiments, multiple lipid molecules may be associated with a polynucleotide molecule. The complex may include 1-100 lipid molecules, 1-1000 lipid molecules, 10-1000 lipid molecules, or 100-10,000 lipid molecules. In certain embodiments, the complex may form a nanoparticle. In a particularly preferred embodiment, the diameter of the nanoparticles ranges from 10-500 nm, more preferably the diameter of the nanoparticles ranges from 10-1200 nm, and most preferably from 50-150 nm.
The nanoparticles may be associated with a targeting agent as described below.

Polynucleotide [00226] The polynucleotide to be complexed, encapsulated by the inventive lipids, or included in a composition with the inventive lipds may be any nucleic acid including but not limited to RNA and DNA. In certain embodiments, the polynucleotide is DNA. In other embodiments, the polynucleotide is RNA. In other embodiments, the polynucleotide is an siRNA. In other embodiments, the polynucleotide is an shRNA. The polynucleotides may be of any size or sequence, and they may be single- or double-stranded. In certain preferred embodiments, the polynucleotide is greater than 100 base pairs long. In certain other preferred embodiments, the polynucleotide is greater than 1000 base pairs long and may be greater than 10,000 base pairs long. The polynucleotide is preferably purified and substantially pure. Preferably, the polynucleotide is greater than 50% pure, more preferably greater than 75% pure, and most preferably greater than 95% pure.
The polynucleotide may be provided by any means known in the art. In certain preferred embodiments, the polynucleotide has been engineered using recombinant techniques (for a more detailed description of these techniques, please see Ausubel et al. Current Protocols in Molecular Biology (John Wiley & Sons, Inc., New York, 1999);
Molecular Cloning: A Laboratory Manual, 2nd Ed., ed. by Sambrook, Fritsch, and Maniatis (Cold Spring Harbor Laboratory Press: 1989); each of which is incorporated herein by reference). The polynucleotide may also be obtained from natural sources and purified from contaminating components found normally in nature. The polynucleotide may also be chemically synthesized in a laboratory. In a preferred embodiment, the polynucleotide is synthesized using standard solid phase chemistry.
[00227] The polynucleotide may be modified by chemical or biological means.
In certain preferred embodiments, these modifications lead to increased stability of the polynucleotide. Modifications include methylation, phosphorylation, end-capping, etc.
[00228] Derivatives of polynucleotides may also be used in the present invention. These derivatives include modifications in the bases, sugars, and/or phosphate linkages of the polynucleotide. Modified bases include, but are not limited to, those found in the following nucleoside analogs: 2-aminoadenosine, 2-thiothymidine, inosine, pyrrolo-pyrimidine, 3-methyl adenosine, 5-methylcytidine, C5-bromouridine, C5-fluorouridine, C5-iodouridine, C5-propynyl-uridine, C5-propynyl-cytidine, C5-methylcytidine, 7-deazaadenosine, 7-deazaguanosine, 8-oxoadenosine, 8-oxoguanosine, 0(6)-methylguanine, and 2-thiocytidine.
Modified sugars include, but are not limited to, 2'-fluororibose, ribose, 2'-deoxyribose, 3'-azido-2',3'-dideoxyribose, 2',3'-dideoxyribose, arabinose (the 2'-epimer of ribose), acyclic sugars, and hexoses. The nucleosides may be strung together by linkages other than the phosphodiester linkage found in naturally occurring DNA and RNA.
Modified linkages include, but are not limited to, phosphorothioate and 5'-N-phosphoramidite linkages. Combinations of the various modifications may be used in a single polynucleotide. These modified polynucleotides may be provided by any means known in the art; however, as will be appreciated by those of skill in this art, the modified polynucleotides are preferably prepared using synthetic chemistry in vitro.
[00229] The polynucleotides to be delivered may be in any form. For example, the polynucleotide may be a circular plasmid, a linearized plasmid, a cosmid, a viral genome, a modified viral genome, an artificial chromosome, etc.
[00230) The polynucleotide may be of any sequence. In certain preferred embodiments, the polynucleotide encodes a protein or peptide. The encoded proteins may be enzymes, structural proteins, receptors, soluble receptors, ion channels, pharmaceutically active proteins, cytokines, interleukins, antibodies, antibody fragments, antigens, coagulation factors, albumin, growth factors, hormones, insulin, etc. The polynucleotide may also comprise regulatory regions to control the expression of a gene. These regulatory regions may include, but are not limited to, promoters, enhancer elements, repressor elements, TATA box, ribosomal binding sites, stop site for transcription, etc. In other particularly preferred embodiments, the polynucleotide is not intended to encode a protein. For example, the polynucleotide may be used to fix an error in the genome of the cell being transfected.
[00231] The polynucleotide may also be provided as an antisense agent or RNA
interference (RNAi) (Fire et al. Nature 391:806-811, 1998; incorporated herein by reference). Antisense therapy is meant to include, e.g., administration or in situ provision of single- or double-stranded oligonucleotides or their derivatives which specifically hybridize, e.g., bind, under cellular conditions, with cellular mRNA
and/or genomic DNA, or mutants thereof, so as to inhibit expression of the encoded protein, e.g., by inhibiting transcription and/or translation (Crooke "Molecular mechanisms of action of antisense drugs" Biochim. Biophys. Acta 1489(l):31-44, 1999; Crooke "Evaluating the mechanism of action of antiproliferative antisense drugs" Antisense Nucleic Acid Drug Dev. 10(2):123-126, discussion 127, 2000;
Methods in Enzyrnology volumes 313-314, 1999; each of which is incorporated herein by reference). The binding may be by conventional base pair complementarity, or, for example, in the case of binding to DNA duplexes, through specific interactions in the major groove of the double helix (i.e., triple helix formation) (Chan et al.
J. Mol. Med.
75(4):267-282, 1997; incorporated herein by reference).
[00232) In a particularly preferred embodiment, the polynucleotide to be delivered comprises a sequence encoding an antigenic peptide or protein.
Nanoparticles containing these polynucleotides can be delivered to an individual to induce an immunologic response sufficient to decrease the chance of a subsequent infection and/or lessen the symptoms associated with such an infection. The polynucleotide of these vaccines may be combined with interleukins, interferon, cytokines, and adjuvants such as cholera toxin, alum, Freund's adjuvant, etc.
A large number of adjuvant compounds are known; a useful compendium of many such compounds is prepared by the National Institutes of Health and can be found on the internet (http:/www.niaid.nih.gov/daids/vaccine/pdf/compendium.pdf, incorporated herein by reference; see also Allison Dev. Biol. Stand. 92:3-11, 1998;
Uiikeless et al.
Annu. Rev. Imnaunol. 6:251-281, 1998; and Phillips et al. Vaccine 10:151-158,1992, each of which is incorporated herein by reference).

[00233) The antigenic protein or peptides encoded by the polynucleotide may be derived from such bacterial organisms as Streptococccus pneumoniae, Haemophilus influenzae, Staphylococcus aureus, Streptococcus pyrogenes, Corynebacterium diphtheriae, Listeria rnonocytogenes, Bacillus anthracis, Clostridium tetani, Clostridium botulinuna, Clostridium perfringens, Neisseria naeningitidis, Neisseria gonorrhoeae, Streptococcus mutans, Pseudomonas aeruginosa, Salmonella typhi, Haemophilus parainf uenzae, BoNdetella pef tussis, Francisella tular=ensis, Yersinia pestis, Vibrio cholerae, Legionella pneumophila, Mycobacterium tuberculosis, Mycobacterium leprae, Treponema pallidum, Leptospirosis interrogans, Borrelia burgdorferi, Canaphylobacterjejuni, and the like; from such viruses as smallpox, influenza A and B, respiratory syncytial virus, parainfluenza, measles, HIV, varicella-zoster, herpes simplex 1 and 2, cytomegalovirus, Epstein-Barr virus, rotavirus, rhinovirus, adenovirus, papillomavirus, poliovirus, mumps, rabies, rubella, coxsackieviruses, equine encephalitis, Japanese encephalitis, yellow fever, Rift Valley fever, hepatitis A, B, C, D, and E virus, and the like; and from such fungal, protozoan, and parasitic organisms such as Cryptococcus neoformans, Histoplasma capsulatuna, Candida albicans, Candida tropicalis, Nocardia asteroides, Rickettsia ricketsii, Rickettsia typhi, Mycoplasma pneumoniae, Chlamydial psittaci, Chlamydial trachomatis, Plasmodium falciparurn, Trypanosoma brucei, Entanzoeba histolytica, Toxoplasma gondii, Trichomonas vaginalis, Schistosoma mansoni, and the like.
Microparticles [002341 The lipids of the present invention may also be used to form drug delivery devices. The inventive lipids may be used to encapsulate agein.ts including polynucleotides, small molecules, proteins, peptides, metals, organometallic compounds, etc. The inventive lipids have several properties that make them particularly suitable in the preparation of drug delivery devices. These include 1) the ability of the lipid to complex and "protect" labile agents; 2) the ability to buffer the pH in the endosome; 3) the ability to act as a "proton sponge" and cause endosomolysis; and 4) the ability to neutralize the charge on negatively charged agents. In a preferred embodiment, the lipids are used to form microparticles containing the agent to be delivered. These microparticles may include other materials such as proteins, carbohydrates, synthetic polymers (e.g., PEG, PLGA), and natural polymers. In a particularly preferred embodiment, the diameter of the microparticles ranges from between 500 nm to 50 micrometers, more preferably from 1 micrometer to 20 micrometers, and most preferably from 1 micrometer to 10 micrometers. In another particularly preferred embodiment, the microparticles range from 1-5 micrometers.

Methods of Preparing Microparticles [00235] The inventive microparticles may be prepared using any method known in this art. These include, but are not limited to, spray drying, single and double emulsion solvent evaporation, solvent extraction, phase separation, simple and complex coacervation, and other methods well known to those of ordinary skill in the art. Particularly preferred methods of preparing the particles are the double emulsion process and spray drying. The conditions used in preparing the microparticles may be altered to yield particles of a desired size or property (e.g., hydrophobicity, hydrophilicity, external morphology, "stickiness", shape, etc.). The method of preparing the particle and the conditions (e.g., solvent, temperature, concentration, air flow rate, etc.) used may also depend on the agent being encapsulated and/or the composition of the matrix.
[00236] Methods developed for making microparticles for delivery of encapsulated agents are described in the literature (for example, please see Doubrow, M., Ed., "Microcapsules and Nanoparticles in Medicine and Pharn7acy," CRC
Press, Boca Raton, 1992; Mathiowitz and Langer, J Controlled Release 5:13-22, 1987;
Mathiowitz et al. Reactive Polymers 6:275-283, 1987; Mathiowitz et al. J.
Appl.
Polymer= Sci. 35:755-774, 1988; each of which is incorporated herein by reference).
[00237] If the particles prepared by any of the above methods have a size range outside of the desired range, the particles can be sized, for example, using a sieve.
The particle may also be coated. In certain embodiments, the particles are coated with a targeting agent. In other embodiments, the particles are coated to achieve deisirable surface properties (e.g., a particular charge).

Micelles and Liposomes [00238] The lipids of the invention may be used to prepare micelles or liposomes. Many techniques for preparing micelles and liposomes are known in the art, and any method may be used with the inventive lipids to make micelles and liposomes. In addition, any agent including polynucleotides, small molecules, proteins, peptides, metals, organometallic compounds, etc. may be included in a micelle or liposome. Micelles and liposomes are particularly useful in delivering hydrophobic agents such as hydrophobic small molecules.

[002391 In certain embodiments, liposomes (lipid vesicles) are formed through spontaneous assembly. In other embodiments, liposomes are formed when thin lipid films or lipid cakes are hydrated and stacks of lipid crystalline bilayers become fluid and swell. The hydrated lipid sheets detach during agitation and self-close to form large, multilamellar vesicles (LMV). This prevents interaction of water with the hydrocarbon core of the bilayers at the edges. Once these particles have formed, reducing the size of the particle can be modified through input of sonic energy (sonication) or mechanical energy (extrusion). See Walde, P. "Preparation of Vesicles (Liposomes)" In Encylopedia of Nanoscience and Nanotechnology; Nalwa, H. S. Ed. American Scientific Publishers: Los Angeles, 2004; Vol. 9, pp. 43-79;
Szoka et al. "Comparative Properties and Methods of Preparation of Lipid Vesicles (Liposomes)" Ann. Rev. Biophys. Bioeng. 9:467-508, 1980; each of which is incorporated herein. The preparation of lipsomes involves preparing the lipid for hydration, hydrating the lipid with agitation, and sizing the vesicles to achieve a homogenous distribution of liposomes. Lipids are first dissolved in an organic solvent to assure a homogeneous mixture of lipids. The solvent is then removed to forin a lipid film. This film is thoroughly dried to remove residual organic solvent by placing the vial or flask on a vaccuum pump overnight. Hydration of the lipid film/cake is accomplished by adding an aqueous medium to the container of dry lipid and agitating the mixture. Disruption of LMV suspensions using sonic energy typically produces small unilamellar vesicles (SUV) with diameters in the range of 15-50 nm. Lipid extrusion is a technique in which a lipid suspension is forced through a polycarbonate filter with a defined pore size to yield particles having a diameter near the pore size of the filter used. Extrusion through filters with 100 nm pores typically yields large, unilamellar vesicles (LUV) with a mean diameter of 120-140 nm.
1002401 In certain embodiments of the invention, liposomes are formed comprising an inventive lipid, PEG-ceramide, cholesterol, and a polynucleotide. In certain embodiments, the polynucleotide is an RNA molecule (e.g., an RNAi molecule). In other embodiments, the polynucleotide is a DNA molecule. In certain embodiments, the lipid is ND98. In other embodiments, the lipid is ND28, ND32, LF94, ND99, ND95, NP103, NP98, ND25, ND20, ND100, NF96, NF103, NF109, NF11, ND24, NF86, NP96, ND36, NF61, NF87, NF95, QG100, NF60, NP100, NF1, NP99, QD99, NF63, LG109, ND103, LF95, QF99, LG100, LF31, LG32, NF109, NF64, LE87, LG77, LG96, ND96, LD31, NG64, ND109, or LG80. In certain embodiments, the amount of lipid in the liposome ranges from 30-80 mol%, preferably 40-70 mol%, more preferably 60-70 mol%. In certain embodiments, the amount of PEG-ceramide in the liposomes ranges from 5-20 mol%, preferably 10-mol%, more preferably approximately 10 mol%. In certain embodiments, the amount of cholesterol in the liposome ranges from 5-25 mol%, preferably 10-20 mol%, more preferably approximately 15 mol%. In certain embodiments, the amount of cholesterol in the liposome is approximately 20 mol%. These liposomes may be prepared using any method known in the art. In certain embodiments (e.g., liposomes containing RNAi molecules), the liposomes are prepared by lipid extrusion.
[00241] Certain lipids can spontaneously self assemble around certain molecules, such as DNA and RNA, to form liposomes. For some applications such as the delivery of polynucleotides, these are preferred. Use of these lipids allows for simple assembly of liposomes without the need for additional steps or devices such as an extruder.
[00242] The following scientific papers described other methods for preparing liposomes and micelles: Narang et al. "Cationic Lipids with Increased DNA
Binding Affinity for Nonviral Gene Transfer in Dividing and Nondividing Cells"
Bioconjugate Cherra. 16:156-68, 2005; Hofland et al. "Formation of stable cationic lipid/DNA
complexes for gene transfer" Proc. Natl. Acad. Sci. USA 93:7305-7309, July 1996;
Byk et al. "Synthesis, Activity, and Structure-Activity Relationship Studies of Novel Cationic Lipids for DNA Transfer" J. Med. Chem. 41(2):224-235, 1998; Wu et al. "Cationic Lipid Polymerization as a Novel Approach for Constructing New DNA
Delivery Agents" Bioconjugate Chem. 12:251-57, 2001; Lukyanov et al. "Micelles from lipid derivatives of water-soluble polymers as delivery systems for poorly soluble drugs" Advanced Drug Delivery Reviews 56:1273-1289, 2004; Tranchant et al. "Physicochemical optimisation of plasmid delivery by cationic lipids" J.
Gene Med. 6:S24-S35, 2004; van Balen et al. "Liposome/Water Lipophilicity: Methods, Information Content, and Pharmaceutical Applications" Medicinal Research Rev.
24(3):299-324, 2004; each of which is incorporated herein by reference.

Agent [00243] The agents to be delivered by the system of the present invention may be therapeutic, diagnostic, or prophylactic agents. Any chemical compound to be administered to an individual may be delivered using the inventive comlexes, nanoparticles, or microparticles. The agent may be a small molecule, organometallic compound, nucleic acid, protein, peptide, polynucleotide, metal, an isotopically labeled chemical compound, drug, vaccine, immunological agent, etc.
[00244] In a preferred embodiment, the agents are organic compounds with pharmaceutical activity. In another embodiment of the invention, the agent is a clinically used drug. In a particularly preferred embodiment, the drug is an antibiotic, anti-viral agent, anesthetic, steroidal agent, anti-inflammatory agent, anti-neoplastic agent, antigen, vaccine, antibody, decongestant, antihypertensive, sedative, birth control agent, progestational agent, anti-cholinergic, analgesic, anti-depressant, anti-psychotic, 0-adrenergic blocking agent, diuretic, cardiovascular active agent, vasoactive agent, non-steroidal anti-inflammatory agent, nutritional agent, etc.
[00245] In a preferred embodiment of the present invention, the agent to be delivered may be a mixture of agents.
[00246] Diagnostic agents include gases; metals; commercially available imaging agents used in positron emissions tomography (PET), computer assisted tomography (CAT), single photon emission computerized tomography, x-ray, fluoroscopy, and magnetic resonance imaging (MRI); and contrast agents.
Examples of suitable materials for use as contrast agents in MRI include gadolinium chelates, as well as iron, magnesium, manganese, copper, and chromium. Examples of materials useful for CAT and x-ray imaging include iodine-based materials.
[00247] Prophylactic agents include, but are not limited to, antibiotics, nutritional supplements, and vaccines. Vaccines may comprise isolated proteins or peptides, inactivated organisms and viruses, dead organisms and viruses, genetically altered organisms or viruses, and cell extracts. Prophylactic agents may be combined with interleukins, interferon, cytokines, and adjuvants such as cholera toxin, alum, Freund's adjuvant, etc. Prophylactic agents include antigens of such bacterial organisms as Streptococccus pneumoniae, Haemophilus influenzae, Staphylococcus aureus, Streptococcus pyrogenes, Coiynebacterium diphtheriae, Listeria monocytogenes, Bacillus anthracis, Clostridium tetani, Clostridium botulinum, Clostridiunz perfringens, Neisseria meningitidis, Neisseria gonorrhoeae, Streptococcus inutans, Pseudomonas aeruginosa, Saltnonella typhi, Haemophilus parainfluenzae, Bordetella pertussis, Francisella tularensis, Yersinia pestis, Vibrio cholerae, Legionella pneumophila, Mycobacterium tuberculosis, Mycobacteriurn leprae, Treponema pallidum, Leptospirosis interrogans, Borrelia burgdorferi, Camphylobacter jejuni, and the like; antigens of such viruses as smallpox, influenza A and B, respiratory syncytial virus, parainfluenza, measles, HIV, varicella-zoster, herpes simplex 1 and 2, cytomegalovirus, Epstein-Barr virus, rotavirus, rhinovirus, adenovirus, papillomavirus, poliovirus, mumps, rabies, rubella, coxsackieviruses, equine encephalitis, Japanese encephalitis, yellow fever, Rift Valley fever, hepatitis A, B, C, D, and E virus, and the like; antigens of fungal, protozoan, and parasitic organisms such as Cryptococcus neofornians, Histoplasnza capsulatum, Candida albicans, Candida tropicalis, Nocardia asteroides, Rickettsia ricketsii, Rickettsia typhi, Mycoplasma pneumoniae, Chlamydial psittaci, Chlamydial trachomatis, Plasrnodiurn falciparum, Trypanosoma brucei, Entamoeba histolytica, Toxoplasma gondii, Trichomonas vaginalis, Schistosoma mansoni, and the like. These antigens may be in the form of whole killed organisms, peptides, proteins, glycoproteins, carbohydrates, or combinations thereof.

Targeting Agents [00248] The inventive complexes, liposomes, micelles, microparticles, and nanoparticles may be modified to include targeting agents since it is often desirable to target a particular cell, collection of cells, or tissue. A variety of targeting agents that direct pharmaceutical compositions to particular cells are known in the art (see, for example, Cotten et al. Methods Enzym. 217:618, 1993; incorporated herein by reference). The targeting agents may be included throughout the particle or may be only on the surface. The targeting agent may be a protein, peptide, carbohydrate, glycoprotein, lipid, small molecule, etc. The targeting agent may be used to target specific cells or tissues or may be used to promote endocytosis or phagocytosis of the particle. Examples of targeting agents include, but are not limited to, antibodies, fragments of antibodies, low-density lipoproteins (LDLs), transferrin, asialycoproteins, gp120 envelope protein of the human immunodeficiency virus (HIV), carbohydrates, receptor ligands, sialic acid, etc. If the targeting agent is included throughout the particle, the targeting agent may be included in the mixture that is used to form the particles. If the targeting agent is only on the surface, the targeting agent may be associated with (i.e., by covalent, hydrophobic, hydrogen bonding, van der Waals, or other interactions) the formed particles using standard chemical techniques.

Pharmaceutical Compositions [00249] Once the complexes, micelles, liposomes, microparticles, or nanoparticles have been prepared, they may be combined with one or more pharmaceutical excipients to form a pharmaceutical composition that is suitable to administer to animals including humans. As would be appreciated by one of skill in this art, the excipients may be chosen based on the route of administration as described below, the agent being delivered, time course of delivery of the agent, etc.
[00250] Pharmaceutical compositions of the present invention and for use in accordance with the present invention may include a pharmaceutically acceptable excipient or carrier. As used herein, the term "pharmaceutically acceptable carrier"
means a non-toxic, inert solid, semi-solid or liquid filler, diluent, encapsulating material or formulation auxiliary of any type. Some examples of materials which can serve as pharmaceutically acceptable carriers are sugars such as lactose, glucose, and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethyl cellulose, ethyl cellulose, and cellulose acetate;
powdered tragacanth; malt; gelatin; talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil; safflower oil;
sesame oil;
olive oil; corn oil and soybean oil; glycols such as propylene glycol; esters such as ethyl oleate and ethyl laurate; agar; detergents such as Tween 80; buffering agents such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethyl alcohol; and phosphate buffer solutions, as well as other non-toxic compatible lubricants such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, releasing agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and-antioxidants can also be present in the composition, according to the judgment of the formulator.
The pharmaceutical compositions of this invention can be administered to humans and/or to animals, orally, rectally, parenterally, intracisternally, intravaginally, intranasally, intraperitoneally, topically (as by powders, creams, ointments, or drops), bucally, or as an oral or nasal spray.
[00251] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups, and elixirs. In addition to the active ingredients (i.e., microparticles, nanoparticles, liposomes, micelles, polynucleotide/lipid complexes), the liquid dosage forms may contain inert diluents commonly used in the art such as, for example, water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (in particular, cottonseed, groundnut, corn, germ, olive, castor, and sesame oils), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof.
Besides inert diluents, the oral compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.
[00252] Injectable preparations, for example, sterile injectable aqueous or oleaginous suspensions may be formulated according to the known art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution, suspension, or emulsion in a nontoxic parenterally acceptable diluent or solvent, for example, as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution, U.S.P. and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose any bland fixed oil can be employed including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid are used in the preparation of injectables. In a particularly preferred embodiment, the particles are suspended in a carrier fluid comprising 1% (w/v) sodium carboxymethyl cellulose and 0.1% (v/v) Tween 80.

[00253] The injectable formulations can be sterilized, for example, by filtration through a bacteria-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved or dispersed in sterile water or other sterile injectable medium prior to use.
[00254] Compositions for rectal or vaginal administration are preferably suppositories which can be prepared by mixing the particles with suitable non-irritating excipients or carriers such as cocoa butter, polyethylene glycol, or a suppository wax which are solid at ambient temperature but liquid at body temperature and therefore melt in the rectum or vaginal cavity and release the microparticles.
[00255] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the particles are mixed with at least one inert, pharmaceutically acceptable excipient or carrier such as sodium citrate or dicalcium phosphate and/or a) fillers or extenders such as starches, lactose, sucrose, glucose, mannitol, and silicic acid, b) binders such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone, sucrose, and acacia, c) humectants such as glycerol, d) disintegrating agents such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate, e) solution retarding agents such as paraffin, f) absorption accelerators such as quaternary ammonium compounds, g) wetting agents such as, for example, cetyl alcohol and glycerol monostearate, h) absorbents such as kaolin and bentonite clay, and i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets, and pills, the dosage form may also comprise buffering agents.
[00256] Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polyethylene glycols and the like.
[00257] The solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings and other coatings well known in the pharmaceutical formulating art. They may optionally contain opacifying agents and can also be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally, in a delayed manner. Examples of embedding compositions which can be used include polymeric substances and waxes.

[00258] Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polyethylene glycols,and the like.
[00259] Dosage forms for topical or transdermal administration of an inventive pharmaceutical composition include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants, or patches. The particles are admixed under sterile conditions with a pharmaceutically acceptable carrier and any needed preservatives or buffers as may be required. Ophthalmic formulation, ear drops, and eye drops are also contemplated as being within the scope of this invention.
[00260] The ointments, pastes, creams, and gels may contain, in addition to the particles of this invention, excipients such as animal and vegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc, and zinc oxide, or mixtures thereof.
[00261] Powders and sprays can contain, in addition to the particles of this invention, excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates, and polyamide powder, or mixtures of these substances. Sprays can additionally contain customary propellants such as chlorofluorohydrocarbons.
[00262] Transdermal patches have the added advantage of providing controlled delivery of a compound to the body. Such dosage forms can be made by dissolving or dispensing the microparticles or nanoparticles in a proper medium. Absorption enhancers can also be used to increase the flux of the compound across the skin. The rate can be controlled by either providing a rate controlling membrane or by dispersing the particles in a polymer matrix or gel.

[00263] These and other aspects of the present invention will be further appreciated upon consideration of the following Examples, which are intended to illustrate certain particular embodiments of the invention but are not intended to limit its scope, as defined by the claims.

Examples Example 1-Preparation and Testing of Amine-containing Lipids [00264] Lipid synthesis. Monomers were purchased from Aldrich (Milwaukee, WI), TCI (Portland, OR), Pfaltz & Bauer (Waterbury, CT), Matrix Scientific (Columbia, SC), Acros-Fisher (Pittsburg, PA), Scientific Polymer (Ontario, NY), Polysciences (Warrington, PA), and Dajac monomer-polymer (Feasterville, PA).
The acrylate and amine monomers were used neat to prepare the lipids. All possible pair wise combinations of amine and acrylate monomers shown in Figure 1 were prepared in sealed vials. The vials were then incubated overnight at approximately 95 C with shaking. The synthesized lipids were used without further purification.
[00265) The molecular weights of the synthesized lipids were determined by mass spectroscopy and compared to predicted molecular weights to confirm synthesis of the lipid. Mass spectrometric data are shown in the table below.
Table 1: Mass-spectrometry data of amine-containing lipids.
Lipid Formula Predicted MW Actual MW
LF1 C38H75NO5 626.572 626.651 LF6 C39H75NO6 654.5667 654.6604 LF7 C40H77N06 668.5824 668.6972 LF10 C37H73NO5 612.5562 612.5917 LF11 C38H75NO5 626.5718 626.6789 LF15 C39H76NO5 638.5723 638.6649 LF17 C44H79NO6 718.598 718.6921 LF20 C37H73NO5 612.5562 612.5946 LF21 C37H73NO5 612.556 612.5959 LG1 C40H79N05 654.6036 654.8644 LG6 C41H79NO6 682.5985 682.8408 LG7 C42H81NO6 696.6142 696.9988 LG10 C39H78NO5 640.588 640.9817 LG13 C40H80NO6 670.5985 670.9156 LG15 C41H79NO5 666.6036 666.9696 LG17 C46H83NO5 746.6298 746.9586 LG20 C39H77N05 640.588 640.9586 LG21 C39H77NO5 640.588 640.9292 LG22 C39H77N05 640.588 640.8809 LG24 C39H77N06 656.5829 656.9402 QF1 C39H78NO5 640.588 640.6866 QF6 C40H78N06 668.5829 668.7032 QF7 C41H80N06 682.5985 682.7867 QF 10 C38H76NO5 626.5723 626.6509 QF11 C39H78NO5 640.588 640.6297 ND25 C33H67N304 570.5204 570.6493 ND36 C36H73N303 596.5725 596.6654 ND75 C36H74N402 595.5885 595.6977 ND87 C37H76N404 641.5939 641.7349 NH32 C47H96N303 750.7451 750.8913 NH36 C48H98N303 764.7608 764.8723 NH60 C45H93N402 735.7455 735.8695 NH86 C48H99N404 795.7666 795.804 NH87 C49H100N404 809.7822 809.8638 Q in the lipid name indicates that the amino groups of the lipid were quaternized using methyl iodide. L indicates lipids prepared from the indicated acrylates and amines. N indicates that the ester functional group of the acrylate has been replaced with an amide group.

[00266] Transfection experiments. 14,000 cos-7 cells (ATCC, Manassas, VA) were seeded into each well of a solid white or clear 96 well plate (Corning-Costar, Kennebunk, ME) and allowed to attached overnight in growth medium, composed of:
500 ml phenol red minus DMEM, 50 ml heat inactivated FBS, 5 ml penicillin/streptomycin (Invitrogen, Carlsbad, CA).
[00267] A small liquot of lipid was tranferred to an Eppendorf tube. Based on the mass of the lipid in the tube, sterile 25 mM sodium acetate buffer was added to each tube to yield a concentration of 60 mg/ml. The resulting mixture was vortexed fro approximately 20 minutes until the lipid was fully dissolved. DNA was prepared based on 300 ng DNA per well of a 96-well plate. 291 g of Lc DNA was dissolved in 9210 l of 25 mM sodium acetate buffer. Aliquots of 30 l of DNA solution were added to each well expect for the last column which was reserved for the Lipo2000 standard. For the last column of the plate, 61 g of DNA was added to 1940 l Optimem. 150 l of inedia/Optimem was added to each well of plates. 50 gl of lipid solution was aliquoted into wells of robot plate. The following amounts were aliquoted to obtain the correct ratios of DNA to lipid:
For 300 ng DNA well:
w/w ratio l of lipid from robot plate l of NaOAc buffer 2.5 5 195 In quadruplicate, 30 l of lipid was aliquoted onto DNA in four rows for each ratio.
For the Lipo2000 control (2.5 w/w ratio to DNA), 152.5 g of Lipo sample was aliquoted into 1847.5 l of Optimem. 30 l of this solution was aliquoted onto DNA
in the Optimem in the last columns of each plate. The plates were incubated for 15-20 minutes, and then 36.5 l of lipid+DNA complexes was transferred into 150 l of media/Optimem, then add to cells. The media was aspirated off the cells, and 105 l of the lipid/DNA/media/Optimem solution was added to the cells. The luciferase assay was performed after 48 hours.

[00268] Luminescence was analyzed using bright-glo assay kits (Promega).
Briefly, 100 gl of bright-glo solution was added to each well of the microtiter plate containing media and cells. Luminescence was measured using a Mithras Luminometer (Berthold, Oak Ridge, TN). In some cases, a neutral density filter (Chroma, Brattleboro, VT) was used to prevent saturation of the luminometer. A
standard curve for Luciferase was generated by titration of Luciferase enzyme (Promega) into growth media in white microtiter plates. Luciferase in ng per well are calculated for each of the lipids at 2.5 w/w, 5 w/w, 10 w/w, 15 w/w, 20 w/w, and 25 w/w lipid to DNA based on the standard curve. These data are shown in the table below. eGFP expression was examined using a Zeiss Aciovert 200 inverted microscope.

Table 2: Luciferase expression (measured in relative light units) as a percentage of that achieved using LipofectamineTM 2000 (ng per well) for lipids at specific lipid/DNA (w/w) ratios using 300 ng Luciferase DNA per well 2.5 w/w 5 w/w 10 w/w 15 w/w 20 w/w 25 w/w QG7 0.4 0.3 0.5 0.3 0.4 0.3 QBI 0.4 0.3 0.4 0.3 0.4 0.4 QF1 0.4 0.3 0.4 0.3 0.3 0.3 QGI 0.4 0.5 0.4 0.4 0.3 0.3 QB77 0.4 0.4 0.4 0.3 0.3 0.3 QF77 0.4 0.4 0.7 1.3 2.7 9.9 QG77 0.5 0.6 1.5 5.6 18.8 29.5 LD90 0.5 0.4 0.4 0.6 0.4 0.4 LE90 0.5 0.3 0.4 0.4 0.4 0.4 LF90 0.5 0.4 0.7 0.4 0.4 0.4 LG90 0.6 0.6 0.4 0.4 0.4 0.4 LB64 0.1 0.1 0.2 0.1 0.1 0.2 LD64 0.1 0.1 0.1 0.1 0.1 0.1 LE64 0.1 0.1 0.1 0.0 0.1 0.1 LF64 0.1 0.1 0.4 0.1 0.2 0.1 LG64 0.2 0.1 0.1 0.1 0.1 0.2 LB31 1.2 0.8 8.3 1.9 1.3 4.2 LD31 44.2 38.7 18.8 11.3 42.8 174.1 LE31 1.0 1.0 0.9 2.5 2.9 9.5 LF31 64.1 78.7 13.4 69.5 97.3 266.8 LG31 19.6 27.6 34.0 8.5 14.3 94.0 LB63 0.1 0.2 0.1 1.0 0.1 0.2 ND28 124.8 116.0 28.9 0.0 0.0 0.0 ND86 0.5 0.2 0.0 0.0 0.0 0.0 ND87 0.0 0.0 0.0 0.0 0.0 0.0 QB6 0.0 0.0 0.0 0.0 0.0 0.0 QF6 0.0 0.0 0.0 0.0 0.0 0.0 QG6 0.0 0.0 0.0 0.0 0.0 0.0 QB7 0.0 0.0 0.0 0.0 0.0 0.0 QF7 0.0 0.0 0.0 0.0 0.0 0.0 LBI 0.1 0.1 0.1 0.1 0.1 0.2 LB6 0.1 0.1 0.1 0.1 0.1 0.1 LB7 0.1 0.1 0.1 0.1 0.1 0.2 LB10 0.1 0.1 0.1 0.1 0.1 0.1 LB11 0.1 0.1 0.1 0.1 0.1 0.1 LB13 0.1 0.1 0.1 0.1 0.1 0.1 LB15 0.1 0.1 0.1 0.1 0.1 0.1 LB1 7 0.1 0.1 0.1 0.1 0.1 0.1 LB20 0.1 0.1 0.4 0.2 0.3 0.7 LB21 0.7 0.8 0.7 0.7 0.7 0.8 LB22 0.3 0.3 0.4 0.4 0.4 0.4 LB24 0.1 0.2 0.4 0.1 0.1 0.1 LB25 0.8 2.4 2.6 3.3 2.3 1.8 LB26 0.1 0.1 0.1 0.1 0.1 0.1 LB28 0.1 0.1 0.1 0.1 0.1 0.1 LB31 0.1 0.1 0.1 0.1 0.1 0.1 LB32 0.1 0.1 0.1 0.1 0.1 0.1 LB33 0.1 0.1 0.1 0.1 0.1 0.1 LB34 0.1 0.1 0.2 0.4 0.1 0.1 LB36 0.1 0.1 0.1 0.1 0.1 0.1 LB38 0.7 0.8 0.7 0.8 0.7 0.8 LB60 0.4 0.4 0.4 0.4 0.4 0.4 LB61 0.1 0.1 0.1 0.1 0.1 0.1 LB62 0.1 0.1 0.1 0.1 0.1 0.1 LB63 0.1 0.1 0.1 0.1 0.1 0.1 LB64 0.1 0.1 0.1 0.1 0.1 0.1 LB70 0.1 0.1 0.1 0.1 0.1 0.1 LB75 0.1 0.1 0.1 0.1 0.1 0.1 LB76 0.1 0.1 0.1 0.1 0.1 0.1 LB77 0.1 0.1 0.1 0.2 0.1 0.3 LB79 0.1 0.1 0.1 0.1 0.1 0.1 LB80 0.7 0.8 0.1 0.2 1.0 1.5 LB81 0.4 0.4 0.2 0.3 0.4 0.5 LF1 0.1 0.1 0.0 0.0 0.1 0.1 LF64 0.1 0.1 0.0 0.0 0.1 0.1 LF7 0.1 0.1 0.0 0.0 0.1 0.1 LF10 0.1 0.1 0.0 0.0 0.1 0.1 LF11 0.1 0.1 0.0 0.0 0.1 0.1 LF13 0.1 0.1 0.0 0.0 0.1 0.1 LF15 0.1 0.1 0.0 0.0 0.1 0.1 LF17 0.1 0.1 0.1 0.1 0.1 0.1 LF20 0.2 0.4 0.3 0.2 0.1 0.1 LF21 0.4 0.4 0.4 0.4 0.4 0.4 LD28 1.7 8.5 LD86 29.6 16.0 LD87 53.9 43.3 LG34 1.4 0.8 LG77 43.5 34.0 LH28 0.2 0.2 QD28 0.1 0.1 QD86 2.0 2.0 QD87 0.5 0.7 LF22 0.1 0.1 0.1 0.1 0.1 0.1 LF24 0.1 0.1 0.1 0.2 0.3 0.3 LF25 0.1 0.2 0.3 0.4 0.9 1.5 LF26 0.1 0.2 0.7 0.1 0.1 0.3 LF28 0.1 0.1 0.1 0.2 0.3 0.4 LF32 0.1 0.1 0.2 0.1 0.4 0.4 LF33 0.1 0.1 0.1 0.1 0.1 0.1 LF34 0.1 0.3 1.1 0.5 0.2 0.3 LF36 0.1 0.1 0.1 0.1 0.1 0.1 LF38 0.8 0.9 0.9 0.9 0.8 1.0 LF60 0.4 0.4 0.5 0.5 0.5 0.5 LF61 0.1 0.1 0.1 0.1 0.1 0.1 LF62 0.1 0.1 0.2 0.1 0.4 0.2 LF63 0.1 0.1 0.1 0.3 0.3 0.4 LF64 0.1 0.1 0.1 0.1 0.1 0.1 LF70 0.1 0.1 0.2 0.3 0.2 0.3 LF75 0.7 1.0 0.7 1.9 1.7 1.7 LF76 1.7 5.6 9.8 24.3 22.2 19.5 LF77 3.7 25.1 28.2 24.1 17.0 22.1 LF79 0.1 0.3 0.3 0.3 0.3 0.3 LF80 2.5 35.1 35.5 34.3 19.9 14.8 LF81 0.5 1.6 1.5 4.9 4.6 4.0 LF82 0.4 0.3 0.7 0.5 0.6 0.8 LF86 21.5 17.9 19.6 21.2 10.5 10.4 LF87 19.4 11.3 30.2 13.3 11.0 10.0 LF90 0.4 0.5 0.3 1.3 1.1 1.5 LF91 0.5 0.5 0.8 1.4 1.3 1.6 LF93 32.0 50.4 15.0 150.7 143.2 171.3 LF94 41.8 37.7 96.3 114.7 99.0 98.6 LF95 15.3 51.3 44.3 71.8 64.6 75.1 LF96 52.4 62.8 79.3 47.7 64.4 36.0 LF98 2.5 7.9 17.8 17.2 9.5 9.9 LF99 32.2 49.8 26.5 10.7 6.0 6.2 LF100 17.6 70.0 69.0 85.9 44.2 50.9 LF103 43.9 11.6 65.4 91.8 61.6 61.4 LF109 16.0 28.3 16.9 21.9 28.7 49.5 LG1 0.0 0.0 0.0 0.0 0.0 0.0 LG64 0.0 0.0 0.0 0.0 0.0 0.0 LG77 0.0 0.0 0.0 0.0 0.0 0.0 LG10 0.0 0.0 0.0 0.0 0.0 0.0 LG11 0.0 0.0 0.0 0.0 0.0 0.0 LG13 0.0 0.0 0.0 0.0 0.0 0.0 LG15 0.7 0.8 0.5 0.6 0.7 0.8 LG17 0.4 0.3 0.1 0.0 0.3 0.3 LG20 2.0 2.5 0.4 0.2 0.1 0.1 LG21 0.1 0.1 0.1 0.2 0.2 0.4 LG22 0.2 0.3 0.4 0.5 0.4 0.7 LG24 2.0 3.3 4.8 11.2 16.4 32.5 LG25 15.9 32.0 43.2 56.9 42.1 63.9 LG26 0.6 13.0 1.9 0.5 0.3 0.4 LG28 0.2 0.5 0.2 0.2 0.1 0.2 LG32 0.2 10.2 1.0 0.7 0.4 0.7 LG33 0.1 0.1 0.1 0.1 0.1 0.1 LG60 0.9 1.0 0.8 0.9 0.8 1.0 LG61 0.4 0.5 0.4 0.4 0.4 0.5 LG63* 0.5 0.5 0.5 0.5 0.5 0.7 LG64* 0.3 0.3 1.8 1.8 1.2 1.6 LG 75* 0.9 1.1 3.1 4.7 3.1 5.3 LG76* 6.2 14.1 21.4 48.6 54.1 92.5 LG79* 0.6 0.4 1.5 1.6 1.4 1.4 LG93* 45.0 43.8 310.5 281.8 185.9 183.8 160A* 0.4 0.4 1.7 1.5 1.1 1.1 160B* 0.8 0.8 1.1 1.1 0.9 0.9 160C* 0.5 0.5 0.7 0.7 0.6 0.8 160D* 0.5 0.3 0.4 0.4 0.3 0.3 160E* 0.6 0.5 0.6 0.6 0.6 0.3 LD109 9.3 18.6 31.1 20.7 7.0 2.0 LD103 11.6 17.2 24.4 27.1 12.3 6.6 LD100 3.9 1.5 12.0 15.3 6.9 1.3 LD99 4.4 12.8 44.6 27.0 6.2 0.8 LD98 0.2 0.7 0.8 1.0 0.8 0.6 LD96 2.3 0.3 1.3 3.6 1.2 0.4 LD95 1.2 19.5 3.6 9.1 9.4 5.9 LD94 1.5 5.9 2.0 8.5 9.2 7.3 LD93 1.8 4.2 3.9 24.6 15.8 10.8 LD91 0.2 0.2 0.2 0.2 0.2 0.2 LD90 0.4 0.4 0.4 0.4 0.5 0.5 LD82 0.1 0.1 0.2 0.3 0.4 0.7 LD81 0.1 0.1 15.2 7.6 5.4 2.3 LD80 0.1 0.1 3.7 6.5 7.1 2.1 LD79 0.1 0.1 0.1 0.2 0.3 0.2 LD77 0.1 0.1 6.9 11.1 6.4 2.9 LD76 0.1 0.1 0.2 0.3 0.2 0.4 LD75 0.1 0.1 0.1 0.2 0.2 0.2 LD70 0.1 0.1 0.6 0.6 0.6 0.6 LD64 0.1 0.1 0.3 0.3 0.3 0.4 LD63 0.7 0.8 0.3 0.3 0.4 0.6 LD62 0.4 0.4 0.4 0.4 0.4 0.5 LG109 16.3 36.0 23.4 37.9 25.7 34.5 LGIOO 21.5 32.7 11.8 18.0 8.6 8.1 LG98 0.7 2.3 13.2 9.8 6.8 7.8 LG96 46.7 82.9 37.3 32.6 14.1 18.5 LG93 3.6 9.2 19.8 37.3 24.0 24.6 LG91 0.2 0.2 0.1 0.2 0.1 0.1 LG90 0.1 0.1 0.0 0.0 0.1 0.0 LG87 14.5 7.5 9.2 10.4 5.9 4.7 LG82 0.2 0.2 0.6 0.8 0.5 1.2 LG81 0.7 0.8 1.2 4.1 14.8 19.9 LG80 2.2 11.9 19.8 31.3 21.9 17.4 LHI 0.0 0.0 0.0 0.0 0.0 0.0 LH6 0.0 0.0 0.0 0.0 0.0 0.0 LH7 0.0 0.0 0.0 0.0 0.0 0.0 LHIO 0.0 0.0 0.0 0.0 0.0 0.0 LHII 0.0 0.0 0.0 0.0 0.0 0.0 LH13 0.0 0.0 0.0 0.0 0.0 0.0 LH15 0.0 0.0 0.0 0.0 0.0 0.0 LH17 0.0 0.0 0.0 0.0 0.0 0.0 LH2O 0.0 0.0 0.0 0.0 0.0 0.0 LH21 0.0 0.0 0.0 0.0 0.0 0.0 LH22 0.0 0.0 0.0 0.0 0.0 0.0 Q in the lipid name indicates that the tertiary amines of the lipid were quaternized using methyl iodide. L indicates lipids prepared from the indicated acrylates and amines. N indicates that the ester functional group of the acrylate has been replaced with an amide group. * indicates 72 hours incubation before bright-glo.

[00269] The table below summarizes the data as a % of the luciferase activity obtained from the use of Lipofectamine 2000. The table indicates the best lipids for transfection.
Table 3 2.5 w/w 5 w/w 10 w/w 15 w/w 20 w/w 25 w/w LD28 1.7 8.5 LD31 44.2 38.7 18.8 11.3 42.8 174 LD77 0.1 0.1 6.9 11.1 6.4 2.9 LD81 0.1 0.1 15.2 7.6 5.4 2.3 LD86 29.6 16.0 LD87 5.3.9 43.3 LD93 1.8 4.2 3.9 24.6 15.8 10.8 LD94 1.5 5.9 2.0 8.5 9.2 7.3 LD95 1.2 19.5 3.6 9.1 9.4 5.9 LD99 4.4 12.8 44.6 27.0 6.2 0.8 LD100 3.9 1.5 12.0 15.3 6.9 1.3 LD103 11.6 17.2 24.4 27.1 12.3 6.6 LD109 9.3 18.6 31.1 20.7 7.0 2.0 LE86 31.1 22.7 22.6 2.5 0.0 0.0 LE87 10.5 9.0 38.4 4.3 0.0 0.0 LE96 13.5 0.5 0.0 0.0 0.0 0.0 LE99 9.9 5.4 13.7 2.7 0.0 0.0 LE103 20.4 22.1 11.8 2.8 0.0 0.0 LE109 1.5 5.3 28.7 18.0 1.8 0.6 LF31 64.,1. 78 7 13.4 6915 , 9,7.3 266 LF76 1.7 5.6 9.8 24.3 22.2 19.5 LF77 3.7 25.1 28.2 24.1 17.0 22.1 LF80 2.5 35.1 35.5 34.3 19.9 14.8 LF86 21.5 17.9 19.6 21.2 10.5 10.4 LF87 19.4 11.3 30.2 13.3 11.0 10.0 LF93 32.0 50.4. 15.0 150.7 1412, 171.3 LF94 41.8 37.7 96':3114.7 , ~99:0', ' 98.6 'LF95 15.3 51:344:3 71'.8 :1 64,6 75.1 LF96 52:4 62.8 79:3 477,64.4 36.0 LF98 2.5 7.9 17.8 17.2 9.5 9.9 LF99 32.2 49.8, 26.5 10.7 6.0 6.2 LFIOO 17.6 70:0 '69:0' ' 859,,,'; 44:2, 501:9' LF103 43.9 11.6 65':4 91.861 :6 61:4 , LF109 16.0 28.3 16.9 21.9 28.7 49:5 LG25 15.9 32.0 43.2 56.9 42.1 63.9 LG31 19.6 27.6 34.0 8.5 14.3 94.0 LG32 0.2 10.2 1.0 0.7 0.4 0.7 LG76* 6.2 14.1 21.4 48.6 54.1 92.5 LG77 43.5 34.0 LG80 2.2 11.9 19.8 31.3 21.9 17.4 LG81 0.7 0.8 1.2 4.1 14.8 19.9 LG87 14.5 7.5 9.2 10.4 5.9 4.7 LG93* 3.6 9.2 19.8 37.3 24.0 24.6 LG96 46.7, 82.9 37.3 32.6 14.1 18.5 LG98 0.7 2.3 13.2 9.8 6.8 7.8 LG100 21.5 32.7 11.8 18.0 8.6 8.1 LG109 16.3 36.0 23.4 37.9 25.7 34.5 LG93 46.0 43:8,, 310,5281.8, 1851.9183 ND28 124.8 10 28.9 0.0 0.0 0.0 QG75 10.0 20.6 34.0 2.7 1.0 0.8 QG76 5.6 16.9 20.7 2.5 0.0 0.0 QG80 1.3 4.9 32.2 684.', 36.2 24.5 QG81 0.7 3.6 41.1 15.5 2.4 1.4 QG82 25.0 24.6 32.6 8.6 2.3 1.4 QG87 89.094 42.4 64.7621 44.1 , QG90 1.1 4.3 7.7 17.4 7.3 5.4 QG91 0.4 3.3 25.9 45.2 18.5 2.8 QG98 2.1 10.3 22.3 14.9 9.3 4.5 QGIOO 11.2 32.1 57.1' 930 .94 QG109 29.9 ~40:6 31':3 52.& 51,:3 466Q in the lipid name indicates that the tertiary amines of the lipid were quaternized using methyl iodide. L indicates lipids prepared from the indicated acrylates and ainines. N indicates that the ester functional group of the acrylate has been replaced with an amide group. * indicates 72 hour incubuation before bright-glo.

Example 2 - Testing of Lipids for RNA Delivery [00270] Reporter-protein knockdown achieved by the top transfecting lipids relative to LipofectamineTM 2000 (where negative values indicate improved knockdown). The assay accounts for toxicity, monitoring expression of both renilla and firefly luciferases, where the latter serves as viability control. For each lipid, 50 ng of siRNA was added per well at specific lipid/RNA w/w ratios (from top to bottom: 2.5, 5, 10, 15).

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?z r ~ LO I' ~~ i O ~ i Example 3- Testing of Lipds for DNA Delivery [00271] Raw values for luciferase expression (relative lights units) of best-transfecting lipids at lipid/DNA (w/w) ratios listed. 300 ng of DNA was added per well.

2.5 w/w 5 w/w 10 w/w 15 w/w [ 33 99~, ~f ND66. . ~___ 8354 12915 ~ ~' ND94 774425 1382667mm,~

['! NQ96_ 29107Q0 42837~

N109219.[484 2087995 NF10 1192'1~8 838233:

f N F 25 2473702 _._ ' 13661~94, ! NF63 114'1469., 865897NF63' J~rv 303171'8 1138429 NF64 384317 2283250 ~ F,70 160574 887500 ~ ~NF91 1993'690_ _ ., 16~065 ~

i NG61 ;,' 1452027 765795 _tiNG64_ 2585784 1972838 N,677 972002 1184771 _ ' -N986 1357355 1521687, '. NG86 151.3703 '1223360~
NG87 823764_õ _,1'005875 j NG95 834849 1208120, Ã

._.-.-_ 4 1;, F NP62 144' 3678 33 188~, , ._.. _ ~_.
NP63 1L I 0415_ 1543275y~ NP63 1388102 i98342 ;

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C7 C7 ~ ~ 0 ~ ~ 0 0 ~ ~ ~ L] ~ L~ 0 L T- LL. IrL L ~ L o' a a F-+
~a ZZZZZZZZZZZZZZ zZZZZZZ

Other Embodiments [00272] The foregoing has been a description of certain non-limiting preferred embodiments of the invention. Those of ordinary skill in the art will appreciate that various changes and modifications to this description may be made without departing from the spirit or scope of the present invention, as defined in the following claims.

Claims (25)

1. A compound of the formula:

wherein V is selected from the group consisting of C=O, C=S, S=O, and SO2;
R1 is selected from the group consisting of hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;

substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstitued, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; -OR A; -C(=O)R A; -CO2R A; -CN; -SCN; -SR A; -SOR A; -SO2R A; -NO2; -N3;
-N(R A)2; -NHC(=O)R A; -NR A C(=O)N(R A)2; -OC(=O)OR A; -OC(=O)R A; -OC(=O)N(R A)2, -NR A C(=O)OR A; or -C(R A)3; wherein each occurrence of R A is independently a hydrogen; a protecting group; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic; substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstitued, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; an acyl moiety;
alkoxy; aryloxy; alkylthio; arylthio; amino, alkylamino, dialkylamino, heteroaryloxy;
or heteroarylthio moiety;
R2 is selected from the group consisting of hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;

substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstitued, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; -OR B; -C(=O)R B; -CO2R B; -CN; -SCN; -SR B; -SOR B; -SO2R B; -NO2; -N3;
-N(R B)2; -NHC(=O)R B; -NR B C(=O)N(R B)2; -OC(=O)OR B; -OC(=O)R B; -OC(=O)N(R B)2; -NR B C(=O)OR B; or -C(R B)3; wherein each occurrence of R B is independently a hydrogen; a protecting group; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic; substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstitued, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; an acyl moiety;
alkoxy; aryloxy; alkylthio; arylthio; amino, alkylamino, dialkylamino, heteroaryloxy;
or heteroarylthio moiety;
wherein R1 and R2 may be taken together to form a cyclic structure;
R3 is selected from the group consisting of hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;

substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstitued, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; -OR c; -C(=O)R C; -CO2R C; -CN; -SCN; -SR C; -SOR C; -SO2R C; -NO2; -N3;
-N(R C)2; -NHC(=O)R C; -NR C C(=O)N(R C)2; -OC(=O)OR C; -OC(=O)R C; -OC(=O)N(R C)2, -NR C C(=O)OR C; or -C(R C)3; wherein each occurrence of R C is independently a hydrogen; a protecting group; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic; substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstitued, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; an acyl moiety;
alkoxy; aryloxy; alkylthio; arylthio; amino, alkylamino, dialkylamino, heteroaryloxy;
or heteroarylthio moiety;
each occurrence of R5 is independently selected from the group consisting of hydrogen and C1-C6 alkyl;
each occurrence of R6 is independently selected from the group consisting of hydrogen and C1-C6 alkyl;
with the proviso that when all occurrences of R5 and R6 are hydrogen, V is C=O, R1 is -OR A, R2 is -OR B, and R1 and R2 are the same, then R3 is not -CH2CH2OR C', wherein R C' is methyl, ethyl, propyl, isopropyl, butyl, s-butyl, isobutyl, t-butyl, pentyl, cyclopentyl, hexyl, cyclohexyl, decyl, methoxymethyl, 2-methoxyethyl, 1-ethoxyethyl, 2-ethoxyethyl, (2-methoxyethoxy)methyl, 2-tetrahydrofuranyl, 2-tetrahydropyranyl, tetrahydrofurfuryl, formyl, acetyl, propionyl, butyryl, isobutyryl, pivaloyl, valeryl, methoxyacetyl, ethoxyacetyl, acetoxyacetyl, 2-formyloxyethyl, 2-acetoxyethyl, 2-oxopropyl, 2-oxobutyl, 2-oxocyclopentyl, 2-oxo-3-tetrahydrofuranyl, 2-oxo-3-tetrahydropyranyl, methoxycarbonyl, ethoxycarbonyl, and t-butoxycarbonyl; and with the proviso that when all occurrences of R5 and R6 are hydrogen, V is C=O, R1 is -OR A, wherein R A is a straight-chain, branched or cyclic alkyl group of 1 to 20 carbon atoms which may contain an ether, carbonyl, or carbonyloxy group;
R2 is -OR B, wherein R B is a straight-chain, branched or cyclic alkyl group of 1 to 20 carbon atoms which may contain an ether, carbonyl, or carbonyloxy group; and R1 and R2 are the same, the R3 is not -CH2CH2OR C", whererin R C" is a straight-chain, branched or cyclic alkyl group of 1 to 20 carbon atoms which may contain an ether, carbonyl, or carbonyloxy group;
and salts thereof.
2. A compound of the formula:

wherein V is selected from the group consisting of C=O, C=S, S=O, and SO2;
R1 is selected from the group consisting of hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;

substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstitued, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; -OR A; -C(=O)R A; -CO2R A; -CN; -SCN; -SR A; -SOR A; -SO2R A; -NO2; -N3;
-N(R A)2; -NHC(=O)R A; -NR A C(=O)N(R A)2; -OC(=O)OR A; -OC(=O)R A; -OC(=O)N(R A)2; -NR A C(=O)OR A; or -C(R A)3; wherein each occurrence of R A is independently a hydrogen; a protecting group; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic; substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstitued, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; an acyl moiety;
alkoxy; aryloxy; alkylthio; arylthio; amino, alkylamino, dialkylamino, heteroaryloxy;
or heteroarylthio moiety;
R2 is selected from the group consisting of hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;

substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstitued, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; -OR B; -C(=O)R B; -CO2R B; -CN; -SCN; -SR B; -SOR B; -SO2R B; -NO2; -N3;
-N(R B)2; -NHC(=O)R B; -NR B C(=O)N(R B)2; -OC(=O)OR B; -OC(=O)R B; -OC(=O)N(R B)2; -NR B C(=O)OR B; or -C(R B)3; wherein each occurrence of R B is independently a hydrogen; a protecting group; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic; substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstitued, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; an acyl moiety;
alkoxy; aryloxy; alkylthio; arylthio; amino, alkylamino, dialkylamino, heteroaryloxy;
or heteroarylthio moiety;

wherein R1 and R2 may be taken together to form a cyclic structure;
R3 is selected from the group consisting of hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;

substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstitued, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; -OR C; -C(=O)R C; -CO2R C; -CN; -SCN; -SR C; -SOR C; -SO2R C; -NO2; -N3;
-N(R C)2; -NHC(=O)R C; -NR C C(=O)N(R C)2; -OC(=O)OR C; -OC(=O)R C; -OC(=O)N(R C)2; -NR C C(=O)OR C; or -C(R C)3; wherein each occurrence of R C is independently a hydrogen; a protecting group; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic; substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstitued, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; an acyl moiety;
alkoxy; aryloxy; alkylthio; arylthio; amino, alkylamino, dialkylamino, heteroaryloxy;
or heteroarylthio moiety;
each occurrence of R5 is independently selected from the group consisting of hydrogen and C1-C6 alkyl;
each occurrence of R6 is independently selected from the group consisting of hydrogen and C1-C6 alkyl;
R7 is hydrogen or C1-C6 aliphatic; and X is an anion;

with the proviso that when all occurrences of R5 and R6 are hydrogen, V is C=O, R1 is -OR A, R2 is -OR B, and R1 and R2 are the same, then R3 is not -CH2CH2OR C', wherein R C' is methyl, ethyl, propyl, isopropyl, butyl, s-butyl, isobutyl, t-butyl, pentyl, cyclopentyl, hexyl, cyclohexyl, decyl, methoxymethyl, 2-methoxyethyl, 1-ethoxyethyl, 2-ethoxyethyl, (2-methoxyethoxy)methyl, 2-tetrahydrofuranyl, 2-tetrahydropyranyl, tetrahydrofurfuryl, formyl, acetyl, propionyl, butyryl, isobutyryl, pivaloyl, valeryl, methoxyacetyl, ethoxyacetyl, acetoxyacetyl, 2-formyloxyethyl, 2-acetoxyethyl, 2-oxopropyl, 2-oxobutyl, 2-oxocyclopentyl, 2-oxo-3-tetrahydrofuranyl, 2-oxo-3-tetrahydropyranyl, methoxycarbonyl, ethoxycarbonyl, and t-butoxycarbonyl; and with the proviso that when all occurrences of R5 and R6 are hydrogen, V is C=O, R1 is -OR A, wherein R A is a straight-chain, branched or cyclic alkyl group of 1 to 20 carbon atoms which may contain an ether, carbonyl, or carbonyloxy group;
R2 is -OR B, wherein R B is a straight-chain, branched or cyclic alkyl group of 1 to 20 carbon atoms which may contain an ether, carbonyl, or carbonyloxy group; and R1 and R2 are the same, the R3 is not -CH2CH2OR C", whererin R C" is a straight-chain, branched or cyclic alkyl group of 1 to 20 carbon atoms which may contain an ether, carbonyl, or carbonyloxy group;
and salts thereof.
3. The compound of claim 2, wherein X is an anion selected from the group consisting of fluoride, chloride, bromide, iodide, sulfate, bisulfate, phosphate, nitrate, acetate, fumarate, oleate, citrate, valerate, maleate, oxalate, isonicotinate, lactate, salicylate, tartrate, tannate, pantothenate, bitartrate, ascorbate, succinate, gentisinate, gluconate, glucaronate, saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, and pamoate (i.e., 1,1'-methylene-bis-(2-hydroxy-3-naphthoate).
4. The compound of claim 2, wherein R7 is C1-C6 alkyl
5. The compound of claim 2, wherein R7 is hydrogen.
6. The compound of claim 2, wherein R7 is methyl
7. The compound of claim 1 or 2, wherein each occurrence of R5 and R6 is hydrogen.
8. The compound of claim 1 or 2, wherein and are the same.
9. The compound of claim 1 or 2, wherein and are the same and are different than R3.
10. The compound of claim 1 or 2, wherein , and R3 are all different.
11. The compound of claim 1 or 2, wherein and are independently selected from the group consisting of:

12. The compound of claim 1 or 2, wherein and are the same and are selected from the group consisting of:

13. The compound of claim 1 or 2, wherein and are the same and are selected from the group consisting of:

14. The compound of claim 1 or 2, wherein and are independently selected from the group consisting of:

15. The compound of claim 1 or 2, wherein and are the same and are selected from the group consisting of:

16. The compound of claim 1 or 2, wherein and are the same and are selected from the group consisting of:
17. The compound of claim 1, wherein is selected from the group consisting of:

18. The compound of claim 1, wherein is selected from the group consisting of:

19. The compound of claim 1, wherein is selected from the group consisting of:

20. The compound of claim 1 of formula:

wherein R3' is C1-6alkyl, hydroxyl, thiol; C1-6alkoxy; amino, C1-6alkylamino, diC1-6alkylamino; carbocyclic moiety; heterocyclic moiety; aryl; or heteroaryl moiety;
n is an integer between 5 and 20, inclusive; and m is an integer between 1 and 10, inclusive; and pharmaceutically acceptable salts thereof.
21. The compound of claim 20, wherein R3' is hydroxyl.
22. The compound of claim 20, wherein R3' is methoxy or ethoxy.
23. The compound of claim 20, wherein R3' is heteroaryl.
24. The compound of claim 20, wherein R3' is histidinyl.
25. The compound of claim 20, wherein R3' is a heterocyclic moiety.

25. The compound of claim 1 selected from the group consisting of:

27. The compound of claim 1 of the formula:

28. The compound of claim 1 of the formula:
wherein V is C=O.

29. The compound of claim 28 of the formula:
wherein R1 is -OR A; and R2 is -OR B.

30. The compound of claim 29, wherein R A and R B are the same.

31. The compound of claim 29, wherein R A and R B are C6-C30 straight chain alkyl groups.

32. The compound of claim 29, wherein R A and R B are C9-C20 straight- chain alkyl groups.

33. The compound of claim 29, wherein R A and R B are C21-C30 straight-chain alkyl groups.

34. The compound of claim 29, wherein R A and R B are C6-C30 straight chain alkenyl groups.

35. The compound of claim 29, wherein R A and R B are C6-C30 straight chain alkynyl groups.

36. The compound of claim 29, wherein R A and R B each comprise at least 4 carbon atoms.

37. The compound of claim 29, wherein R A and R B each comprise at least 5 carbon atoms.

38. The compound of claim 29, wherein R A and R B each comprise at least 6 carbon atoms.

39. The compound of claim 29, wherein R A and R B are independently selected from the group consisting of a hydrogen; a protecting group; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;

substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstitued, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; an acyl moiety; alkoxy; aryloxy; alkylthio; arylthio; amino, alkylamino, dialkylamino, heteroaryloxy; or heteroarylthio moiety.

40. The compound of claim 29, wherein R A and R B are cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic moities.

41. The compound of claim 29, wherein R A and R B are the same.

42. The compound of claim 29, wherein R A and R B are acyclic, substituted or unsubstituted, branched or unbranched aliphatic moieties.

43. The compound of claim 29, wherein R A and R B are acyclic, unsubstituted, unbranched aliphatic moeities.

44. The compound of claim 29, wherein R A and R B are alkyl groups.

45. The compound of claim 29, wherein R A and R B are C1-C30 alkyl groups.
46. The compound of claim 29, wherein R A and R B are C5-C20 alkyl groups.
47. The compound of claim 29, wherein R A and R B are C5-C12 alkyl groups.
48. The compound of claim 29, wherein R A and R B are polyethylene glycol groups.

49. The compound of claim 29, wherein R3 is a cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic moiety; a cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic moiety; a substituted or unsubstitued, branched or unbranched aryl moiety; or a substituted or unsubstituted, branched or unbranched heteroaryl moiety.

50. The compound of claim 29, wherein R3 is not 51. The compound of claim 28 of the formula:

wherein R1 is -N(R A)2; and R2 is N(R B)2.

52. The compound of claim 28 of the formula:

wherein R1 is -NHR A; and R2 is -NHR B.

53. A compound of the formula:

wherein A is selected from the group consisting of cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic; substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstitued, branched or unbranched aryl; and substituted or unsubstituted, branched or unbranched heteroaryl;
V is selected from the group consisting of C=O, C=S, S=O, and SO2;
R1 is selected from the group consisting of hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;

substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstitued, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; -OR A; -C(=O)R A; -CO2R A; -CN; -SCN; -SR A; -SOR A; -SO2R A; -NO2; -N3;
-N(R A)2; -NHC(=O)R A; -NR A C(=O)N(R A)2; -OC(=O)OR A; -OC(=O)R A; -OC(=O)N(R A)2; -NR A C(=O)OR A; and -C(R A)3; wherein each occurrence of R A
is independently a hydrogen; a protecting group; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic; substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstitued, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; an acyl moiety;
alkoxy; aryloxy; alkylthio; arylthio; amino, alkylamino, dialkylamino, heteroaryloxy;
or heteroarylthio moiety;
R2 is selected from the group consisting of hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;

substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstitued, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; -OR B; -C(=O)R B; -CO2R B; -CN; -SCN; -SR B; -SOR B; -SO2R B; -NO2; -N3;
-N(R B)2; -NHC(=O)R B; -NR B C(=O)N(R B)2; -OC(=O)OR B; -OC(=O)R B; -OC(=O)N(R B)2; -NR B C(=O)OR B; or -C(R B)3; wherein each occurrence of R B is independently a hydrogen; a protecting group; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic; substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstitued, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; an acyl moiety;
alkoxy; aryloxy; alkylthio; arylthio; amino, alkylamino, dialkylamino, heteroaryloxy;
or heteroarylthio moiety;
wherein R1 and R2 may be taken together to form a cyclic structure;
R3 is selected from the group consisting of hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;

substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstitued, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; -OR C; -C(=O)R C; -CO2R C; -CN; -SCN; -SR C; -SOR C; -SO2R C; -NO2; -N3;
-N(R C)2; -NHC(=O)R C; -NR C C(=O)N(R C)2; -OC(=O)OR C; -OC(=O)R C; -OC(=O)N(R C)2; -NR C C(=O)OR C; or -C(R C)3; wherein each occurrence of R C is independently a hydrogen; a protecting group; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic; substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstitued, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; an acyl moiety;
alkoxy; aryloxy; alkylthio; arylthio; amino, alkylamino, dialkylamino, heteroaryloxy;
or heteroarylthio moiety;
R4 is selected from the group consisting of hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;

substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstitued, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; -OR D; -C(=O)R D; -CO2R D; -CN; -SCN; -SR D; -SOR D; -SO2R D; -NO2; -N3;
-N(R D)2; -NHC(=O)R D; -NR C C(=O)N(R D)2; -OC(=O)OR D; -OC(=O)R D; -OC(=O)N(R D)2; -NR C C(=O)OR D; or -C(R D)3; wherein each occurrence of R D is independently a hydrogen; a protecting group; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic; substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstitued, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; an acyl moiety;
alkoxy; aryloxy; alkylthio; arylthio; amino, alkylamino, dialkylamino, heteroaryloxy;
or heteroarylthio moiety;
wherein R3 and R4 may be taken together to form a cyclic structure;
and salts thereof.

54. A compound of the formula:

wherein A is selected from the group consisting of cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic; substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstitued, branched or unbranched aryl; and substituted or unsubstituted, branched or unbranched heteroaryl;
V is selected from the group consisting of C=O, C=S, S=O, and SO2;
R1 is selected from the group consisting of hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;

substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstitued, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; -OR A; -C(=O)R A; -CO2R A; -CN; -SCN; -SR A; -SOR A; -SO2R A; -NO2; -N3;
-N(R A)2; -NHC(=O)R A; -NR A C(=O)N(R A)2; -OC(=O)OR A; -OC(=O)R A; -OC(=O)N(R A)2; -NR A C(=O)OR A; and -C(R A)3; wherein each occurrence of R A
is independently a hydrogen; a protecting group; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic; substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstitued, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; an acyl moiety;
alkoxy; aryloxy; alkylthio; arylthio; amino, alkylamino, dialkylamino, heteroaryloxy;
or heteroarylthio moiety;
R2 is selected from the group consisting of hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;

substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstitued, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; -OR B; -C(=O)R B; -CO2R B; -CN; -SCN; -SR B; -SOR B; -SO2R B; -NO2; -N3;
-N(R B)2; -NHC(=O)R B; -NR B C(=O)N(R B)2; -OC(=O)OR B; -OC(=O)R B; -OC(=O)N(R B)2; -NR B C(=O)OR B; or -C(R B)3; wherein each occurrence of R B
is independently a hydrogen; a protecting group; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic; substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstitued, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; an acyl moiety;

alkoxy; aryloxy; alkylthio; arylthio; amino, alkylamino, dialkylamino, heteroaryloxy;
or heteroarylthio moiety;

wherein R1 and R2 may be taken together to form a cyclic structure;
R3 is selected from the group consisting of hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;

substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstitued, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; -OR C; -C(=O)R C; -CO2R C; -CN; -SCN; -SR C; -SOR C; -SO2R C; -NO2; -N3;
-N(R c)2; -NHC(=O)R c; -NR c C(=O)N(R c)2; -OC(=O)OR c; -OC(=O)R c; -OC(=O)N(R c)2; -NR c C(=O)OR c; or -C(R c)3; wherein each occurrence of R c is independently a hydrogen; a protecting group; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic; substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstitued, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; an acyl moiety;
alkoxy; aryloxy; alkylthio; arylthio; amino, alkylamino, dialkylamino, heteroaryloxy;
or heteroarylthio moiety;
R4 is selected from the group consisting of hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;

substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstitued, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; -OR D; -C(=O)R D; -CO2R D; -CN; -SCN; -SR D; -SOR D; -SO2R D; -NO2; -N3;
-N(R D)2; -NHC(=O)R D; -NR c C(=O)N(R D)2; -OC(=O)OR D; -OC(=O)R D; -OC(=O)N(R D)2; -NR c C(=O)OR D; or -C(R D)3; wherein each occurrence of R D is independently a hydrogen; a protecting group; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic; substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstitued, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; an acyl moiety;
alkoxy; aryloxy; alkylthio; arylthio; amino, alkylamino, dialkylamino, heteroaryloxy;

or heteroarylthio moiety;
wherein R3 and R4 may be taken together to form a cyclic structure;
each occurrence of R7 is C1-C6 aliphatic;
X is an anion; and salts thereof.

55. The compound of claim 54, wherein X is an anion selected from the group consisting of fluoride, chloride, bromide, iodide, sulfate, bisulfate, phosphate, nitrate, acetate, fumarate, oleate, citrate, valerate, maleate, oxalate, isonicotinate, lactate, salicylate, tartrate, tannate, pantothenate, bitartrate, ascorbate, succinate, gentisinate, gluconate, glucaronate, saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, and pamoate (i.e., 1,1'-methylene-bis-(2-hydroxy-3-naphthoate).

56. The compound of claim 54, wherein R7 is hydrogen.

57. The compound of claim 54, wherein R7 is C1-C6 alkyl.

58. The compound of claim 54, wherein R7 is methyl.

59. The compound of claim 53, wherein A is a cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic moiety; or a cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic moiety.

60. The compound of claim 53, wherein A is an acyclic, unsubstituted, unbranched aliphatic moiety.

61. The compound of claim 53, wherein A is an acyclic, unsubstituted, unbranched alkyl group.

62. The compound of claim 53, wherein A is an acyclic, unsubstituted, unbranched C1-C6 alkyl group.

63. The compound of claim 53, wherein A is an acyclic, unsubstituted, unbranched heteroaliphatic moiety.

64. The compound of claim 53, wherein A is a polyethylene glycol moiety.

65. The compound of claim 53, wherein R3 and R4 are the same.

66. The compound of claim 53, wherein R3 and R4 are C1-C6 alkyl group.

67. The compound of claim 53, wherein R3 or R4 is 68. The compound of claim 53, wherein is selected from the group consisting of:

69. The compound of claim 53, wherein is selected from the group consisting of:

70. The compound of claim 53, wherein is selected from the group consisting of:

71. The compound of claim 53, wherein is selected from the group consisting of:

72. A compound of formula:

wherein A is selected from the group consisting of cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic; substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; and substituted or unsubstituted, branched or unbranched heteroaryl;
V is selected from the group consisting of C=O, C=S, S=O, and SO2;
R1 is selected from the group consisting of hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;

substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; -OR A; -C(=O)R A; -CO2R A; -CN; -SCN; -SR A; -SOR A; -SO2R A; -NO2; -N3; -N(R A)2; -NHC(=O)R A; -NR A C(=O)N(R A)2; -OC(=O)OR A; -OC(=O)R A; -OC(=O)N(R A)2; -NR A C(=O)OR A; and -C(R A)3; wherein each occurrence of R A is independently a hydrogen; a protecting group; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;
substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; an acyl moiety; alkoxy; aryloxy; alkylthio; arylthio; amino, alkylamino, dialkylamino, heteroaryloxy; or heteroarylthio moiety;
R2 is selected from the group consisting of hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;

substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; -OR B; -C(=O)R B; -CO2R B; -CN; -SCN; -SR B; -SOR B; -SO2R B; -NO2; -N3; -N(R B)2; -NHC(=O)R B; -NR B C(=O)N(R B)2; -OC(=O)OR B; -OC(=O)R B; -OC(=O)N(R B)2; NR B C(=O)OR B; or -C(R B)3; wherein each occurrence of R B is independently a hydrogen; a protecting group; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;
substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; an acyl moiety; alkoxy; aryloxy; alkylthio; arylthio; amino, alkylamino, dialkylamino, heteroaryloxy; or heteroarylthio moiety;
wherein R1 and R2 may be taken together to form a cyclic structure;
R3 is selected from the group consisting of hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;

substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; -OR C; -C(=O)R c; -CO2R C; -CN; -SCN; -SR c; -SOR c; -SO2R c; -NO2; -N3; -N(R c)2; -NHC(=O)R c; -NR c C(=O)N(R c)2; -OC(=O)OR c; -OC(=O)R c; -OC(=O)N(R c)2; -NR c C(=O)OR c; or -C(R c)3; wherein each occurrence of R c is independently a hydrogen; a protecting group; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;
substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; an acyl moiety; alkoxy; aryloxy; alkylthio; arylthio; amino, alkylamino, dialkylamino, heteroaryloxy; or heteroarylthio moiety;
R4 is selected from the group consisting of hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;

substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; -OR D; -C(=O)R D; -CO2R D; -CN; -SCN; -SR D; -SOR D; -SO2R D; -NO2; -N3; -N(R D)2; -NHC(=O)R D; -NR c C(=O)N(R D)2; -OC(=O)OR D; -OC(=O)R D; -OC(=O)N(R D)2; -NR c C(=O)OR D; or -C(R D)3; wherein each occurrence of R D is independently a hydrogen; a protecting group; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;
substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; an acyl moiety; alkoxy; aryloxy; alkylthio; arylthio; amino, alkylamino, dialkylamino, heteroaryloxy; or heteroarylthio moiety;
wherein R3 and R4 may be taken together to form a cyclic structure;
each occurrence of R5 is independently selected from the group consisting of hydrogen and C1-C6 alkyl;
each occurrence of R6 is independently selected from the group consisting of hydrogen and C1-C6 alkyl;
R7 is selected from the group consisting of hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;

substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; -OR G; -C(=O)R G; -CO2R G; -CN; -SCN; -SR G; -SOR G; -SO2R G; -NO2; -N3; -N(R G)2; -NHC(=O)R G; -NR G C(=O)N(R G)2; -OC(=O)OR G; -OC(=O)R G; -OC(=O)N(R G)2; -NR G C(=O)OR G; and -C(R G)3; wherein each occurrence of R G is independently a hydrogen; a protecting group; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;
substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; an acyl moiety; alkoxy; aryloxy; alkylthio; arylthio; amino, alkylamino, dialkylamino, heteroaryloxy; or heteroarylthio moiety and salts thereof.

73. A compounds of the formula:

wherein A is selected from the group consisting of cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic; substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; and substituted or unsubstituted, branched or unbranched heteroaryl;
V is selected from the group consisting of C=O, C=S, S=O, and SO2;
R1 is selected from the group consisting of hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;

substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; -OR A; -C(=O)R A; -CO2R A; -CN; -SCN; -SR A; -SOR A; -SO2R A; NO2; -N3; -N(R A)2; -NHC(=O)R A; -NR A C(=O)N(R A)2; -OC(=O)OR A; -OC(=O)R A; -OC(=O)N(R A)2; -NR A C(=O)OR A; and -C(R A)3; wherein each occurrence of R A is independently a hydrogen; a protecting group; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;
substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; an acyl moiety; alkoxy; aryloxy; alkylthio; arylthio; amino, alkylamino, dialkylamino, heteroaryloxy; or heteroarylthio moiety;
R2 is selected from the group consisting of hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;

substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; -OR B; -C(=O)R B; -CO2R B; -CN; -SCN; -SR B; -SOR B; -SO2R B; -NO2; -N3; -N(R B)2; -NHC(=O)R B; -NR B C(=O)N(R B)2; -OC(=O)OR B; -OC(=O)R B; -OC(=O)N(R B)2; -NR B C(=O)OR B; or -C(R B)3; wherein each occurrence of R B is independently a hydrogen; a protecting group; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;
substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; an acyl moiety; alkoxy; aryloxy; alkylthio; arylthio; amino, alkylamino, dialkylamino, heteroaryloxy; or heteroarylthio moiety;
wherein R1 and R2 may be taken together to form a cyclic structure;
R3 is selected from the group consisting of hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;

substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; -OR C; -C(=O)R c; -CO2R c; -CN; -SCN; -SR c; -SOR c; -SO2R c; -NO2; -N3; -N(R C)2; -NHC(=O)R c; -NR C C(=O)N(R c)2; -OC(=O)OR C; -OC(=O)R c; -OC(=O)N(R C)2; -NR C C(=O)OR C; or -C(R C)3; wherein each occurrence of R c is independently a hydrogen; a protecting group; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;
substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; an acyl moiety; alkoxy; aryloxy; alkylthio; arylthio; amino, alkylamino, dialkylamino, heteroaryloxy; or heteroarylthio moiety;
R4 is selected from the group consisting of hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;

substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; -OR D; -C(=O)R D; -CO2R D; -CN; -SCN; -SR D; -SOR D; -SO2R D; -NO2; -N3; -N(R D)2; -NHC(=O)R D; -NR c C(=O)N(R D)2; -OC(=O)OR D; -OC(=O)R D; -OC(=O)N(R D)2; -NR C C(=O)OR D; or -C(R D)3; wherein each occurrence of R D is independently a hydrogen; a protecting group; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;
substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; an acyl moiety; alkoxy; aryloxy; alkylthio; arylthio; amino, alkylamino, dialkylamino, heteroaryloxy; or heteroarylthio moiety;
wherein R3 and R4 may be taken together to form a cyclic structure;
each occurrence of R5 is independently selected from the group consisting of hydrogen and C1-C6 alkyl;
each occurrence of R6 is independently selected from the group consisting of hydrogen and C1-C6 alkyl;
R7 is selected from the group consisting of hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;

substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; -OR G; -C(=O)R G; -CO2R G; -CN; -SCN; -SR G; -SOR G; -SO2R G; -NO2; -N3; -N(R G)2; -NHC(=O)R G; -NR G C(=O)N(R G)2; -OC(=O)OR G; -OC(=O)R G; -OC(=O)N(R G)2; -NR G C(=O)OR G; and -C(R G)3; wherein each occurrence of R G is independently a hydrogen; a protecting group; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;
substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; an acyl moiety; alkoxy; aryloxy; alkylthio; arylthio; amino, alkylamino, dialkylamino, heteroaryloxy; or heteroarylthio moiety;
each occurrence of R8 is hydrogen or C1-C6 aliphatic;
each dashed line represents a bond or the absence of a bond, wherein when the dashed line represents a bond, the attached nitrogen is positively charged;
and X is any anion.

74. The compound of claim 73, wherein each occurrence of R8 is hydrogen or methyl.

75. The compound of claim 72, wherein is selected from the group 76. The compound of claim 72 of the formula:

wherein each occurrence of x is an integer between 1 and 10, inclusive;
y is an integer between 0 and 10, inclusive;

each occurrence of R7 is hydrogen or R1 is selected from the group consisting of hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;

substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; -OR A; -C(=O)R A; -CO2R A; -CN; -SCN; -SR A; -SOR A; -SO2R A; -NO2; -N3; -N(R A)2; -NHC(=O)R A; -NR A C(=O)N(R A)2; -OC(=O)OR A; -OC(=O)R A; -OC(=O)N(R A)2; -NR A C(=O)OR A; and -C(R A)3; wherein each occurrence of R A is independently a hydrogen; a protecting group; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;
substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstituted, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; an acyl moiety; alkoxy; aryloxy; alkylthio; arylthio; amino, alkylamino, dialkylamino, heteroaryloxy; or heteroarylthio moiety; and salts thereof.

77. The compound of claim 76, wherein x is an integer between 1 and 6, inclusive.
78. The compound of claim 76, wherein y is an integer between 1 and 3, inclusive.
79. The compound of claim 76, whereinR1 is -OR A.

80. The compound of claim 76, wherein R1 is -ONHR A.
81. The compound of claim 76, wherein R7 is hydrogen or 82. The compound of claim 76, wherein R7 is hydrogen or 83. A composition prepared by the method of reacting an amine of one of the formula (1-117):

with an acrylate or acrylamide of formula:

84. The composition of claim 83, wherein the amine is amine 1, 10, 11, 20, 24, 25, 28, 31, 32, 36, 60, 61, 62, 63, 64, 66, 77, 80, 86, 87, 91, 94, 95, 96, 98, 99, 100, 103, or 109.

85. The composition of claim 83, wherein an amine is reacted with an acrylate.

86. The composition of claim 83, wherein an amine is reacted with an acrylamide.
87. The composition of claim 83, wherein the acrylate is acrylate LD, LE, LF, or LG.

88. The composition of claim 83, wherein the acrylamide is acrylamide ND, NF, NG, or NP.

89. The composition of claim 83, wherein amine 98 is reacted with acrylamide ND; or amine 94 is reacted with acrylamide ND; or amine 20 is reacted with acrylamide ND; or amine 24 is reacted with acrylamide ND; or amine 25 is reacted with acrylamide ND; or amine 28 is reacted with acrylamide ND; or amine 36 is reacted with acrylamide ND; or amine 95 is reacted with acrylamide ND; or amine 96 is reacted with acrylamide ND; or amine 99 is reacted with acrylamide ND; or amine 100 is reacted with acrylamide ND; or amine 32 is reacted with acrylamide ND;
or amine 103 is reacted with acrylamide ND; or amine 109 is reacted with acrylamide ND; or amine 1 is reacted with acrylamide NF; or amine 10 is reacted with acrylamide NF; or amine 11 is reacted with acrylamide NF; or amine 20 is reacted with acrylamide NF; or amine 25 is reacted with acrylamide NF; or amine 63 is reacted with acrylamide NF; or amine 60 is reacted with acrylamide NF; or amine 61 is reacted with acrylamide NF; or amine 64 is reacted with acrylamide NF; or amine 86 is reacted with acrylamide NF; or amine 87 is reacted with acrylamide NF;
or amine 103 is reacted with acrylamide NF; or amine 91 is reacted with acrylamide NF;
or amine 95 is reacted with acrylamide NF; or amine 96 is reacted with acrylamide NF; or amine 109 is reacted with acrylamide NF; or amine 61 is reacted with acrylamide NG; or amine 64 is reacted with acrylamide NG; or amine 77 is reacted with acrylamideNG; or amine 86 is reacted with acrylamide NG; or amine 87 is reacted with acrylamide NG; or amine 95 is reacted with acrylamide NG; or amine 62 is reacted with acrylamide NP; or amine 63 is reacted with acrylamine NP, or amine 86 is reacted with acrylamide NP; or amine 87 is reacted with acrylamide NP;
or amine 96 is reacted with acrylamide NP; or amine 98 is reacted with acrylamide NP;
or amine 99 is reacted with acrylamide NP; or amine 100 is reacted with acrylamide NP; or amine 103 is reacted with acrylamide NP; or amine 31 is reacted with acrylate LD; or amine 99 is reacted with acrylate LD; or amine 87 is reacted with acrylate LE;
or amine 31 is reacted with acrylate LF; or amine 95 is reacted with acrylate LF; or amine 94 is reacted with acrylate LF; or amine 99 is reacted with acrylate LF;
or amine 32 is reacted with acrylate LG; or amine 77 is reacted with acrylate LG;
or amine 80 is rected with acrylate LG; or amine 96 is reacted with acrylate LG;
or amine 100 is reacted with acrylate LG; or amine 109 is reacted with acrylate LG; or amine 64 is reacted with acrylate NG; or amine 32 is reacted with acrylate LG;
or amine 31 is reacted with acrylate LG.

90. The composition of claim 83, wherein amine 98 is reacted with acrylamide ND.

91. The composition of claim 83, wherein the composition is a mixture of lipids 92. A compound of formula:

93. The compound of claim 92 of formula:

95. A composition comprising one or more of the compounds of formula:

96. A microparticle comprising a compound of claim 1 and an agent to be delivered.

97. A liposome comprising a compound of claim 1 and an agent to be delivered.

98. The liposome of claim 97, wherein the agent is a polynucleotide.

99. The liposome of claim 97, hwerein the agent is RNA.

100. The liposome of claim 97, wherein the agent is a drug.

101. The liposome of claim 97, wherein the agent is a protein or peptide.

102. The liposome of claim 97, wherein the agent is a small molecule.

103. The liposome of claim 97, wherein the agent is a gas.

104. The liposome of claim 97, wherein the liposome ranges in size from 20 nanometers to 2000 nanometers.

105. The liposome of claim 97 further comprising cholesterol.

106. The liposome of claim 97 further comprising PEG-ceramide.

107. The liposome of claim 97 comprising a compound of claim 1;

an agent, wherein the agent is an RNA;
cholesterol; and PEG-ceramide.

108. A micelle comprising a compound of claim 1 and an agent to be delivered.

109. A pharmaceutical composition comprising a compound of claim 1 and pharmaceutical agent.

110. The pharmaceutical composition of claim 109, wherein the pharmaceutical agent is selected from the groups consisting of polynucleotides, proteins, peptides, and small molecule drugs.

111. The pharmaceutical composition of claim 109, wherein the pharmaceutical agent is a polynucleotide.

112. A method of preparing a compound of claim 1, the method comprising steps of:
reacting one or more equivalents of a compound of formula:

wherein R1 is selected from the group consisting of hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;

substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstitued, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; -OR A; -C(=O)R A; -CO2R A; -CN; -SCN; -SR A; -SOR A; -SO2R A; -NO2; -N3;
-N(R A)2; -NHC(=O)R A; -NR A C(=O)N(R A)2; -OC(=O)OR A; -OC(=O)R A; -OC(=O)N(R A)2; -NR A C(=O)OR A; or -C(R A)3; wherein each occurrence of R A is independently a hydrogen; a protecting group; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic; substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstitued, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; an acyl moiety;
alkoxy; aryloxy; alkylthio; arylthio; amino, alkylamino, dialkylamino, heteroaryloxy;
or heteroarylthio moiety; and R5 is independently selected from the group consisting of hydrogen and C1-C6 alkyl;
with an amine of formula:

wherein R3 is selected from the group consisting of hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;

substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstitued, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; -OR C; -C(=O)R C; -CO2R C; -CN; -SCN; -SR C; -SOR C; -SO2R C; -NO2; -N3;
-N(R C)2; -NHC(=O)R C; -NR C C(=O)N(R C)2; -OC(=O)OR C; -OC(=O)R C; -OC(=O)N(R C)2; -NR C C(=O)OR C; or -C(R C)3; wherein each occurrence of R C is independently a hydrogen; a protecting group; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic; substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstitued, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; an acyl moiety;
alkoxy; aryloxy; alkylthio; arylthio; amino, alkylamino, dialkylamino, heteroaryloxy;
or heteroarylthio moiety;
to form a compound of formula:

113. A method of preparing a compound of claim 53, the method comprising steps of:
reacting a compound of formula:

wherein R1 is selected from the group consisting of hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;

substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstitued, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; -OR A; -C(=O)R A; -CO2R A; -CN; -SCN; -SR A; -SOR A; -SO2R A; -NO2; -N3;
-N(R A)2; -NHC(=O)R A; -NR A C(=O)N(R A)2; -OC(=O)OR A; -OC(=O)R A; -OC(=O)N(R A)2; -NR A C(=O)OR A; or -C(R A)3; wherein each occurrence of R A is independently a hydrogen; a protecting group; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic; substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstitued, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; an acyl moiety;
alkoxy; aryloxy; alkylthio; arylthio; amino, alkylamino, dialkylamino, heteroaryloxy;
or heteroarylthio moiety; and R5 is independently selected from the group consisting of hydrogen and C1-C6 alkyl;
with an amine of formula:

wherein A is selected from the group consisting of cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic; substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstitued, branched or unbranched aryl; and substituted or unsubstituted, branched or unbranched heteroaryl;
R3 is selected from the group consisting of hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;

substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstitued, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; -OR C; -C(=O)R C; -CO2R C; -CN; -SCN; -SR C; -SOR C; -SO2R C; -NO2; -N3;
-N(R C)2; -NHC(=O)R C; -NR C C(=O)N(R C)2; -OC(=O)OR C; -OC(=O)R C; -OC(=O)N(R C)2; -NR C C(=O)OR C; or -C(R C)3; wherein each occurrence of R C is independently a hydrogen; a protecting group; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic; substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstitued, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; an acyl moiety;
alkoxy; aryloxy; alkylthio; arylthio; amino, alkylamino, dialkylamino, heteroaryloxy;
or heteroarylthio moiety;
R4 is selected from the group consisting of hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;

substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstitued, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; -OR D; -C(=O)R D; -CO2R D; -CN; -SCN; -SR D; -SOR D; -SO2R D; -NO2; -N3;
-N(R D)2; -NHC(=O)R D; -NR C C(=O)N(R D)2; -OC(=O)OR D; -OC(=O)R D; -OC(=O)N(R D)2; -NR C C(=O)OR D; or -C(R D)3; wherein each occurrence of R D is independently a hydrogen; a protecting group; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic; substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstitued, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; an acyl moiety;
alkoxy; aryloxy; alkylthio; arylthio; amino, alkylamino, dialkylamino, heteroaryloxy;
or heteroarylthio moiety;
wherein R3 and R4 may be taken together to form a cyclic structure to form a compound of formula:

114. The method of claim 113, wherein wherein is selected from the group consisting of:

115. The method of claim 112, wherein R1 is -OR A.

116. The method of claim 115, wherein -OR A is 117. The method of claim 112, wherein R1 is -NHR A.

118. The method of claim 117, wherein -NHR A is 119. The method of claim 112, wherein is selected from the group consisting of:

120. The method of claim 112, wherein the step of reacting is performed with no solvent present.

121. The method of claim 112, wherein the step of reacting is performed in the presence of an aprotic solvent.

122. The method of claim 112, wherein the solvent is tetrahydrofuran, diethyl ether, ethyl acetate, DMSO, or DMF.

123. The method of claim 112, wherein the step of reacting is performed at a temperature ranging from 25 °C to 100 °C.

124. The method of claim 123, wherein the step of reacting is performed at approximately 95°C.

125. The method of claim 112 further comprising the step of purifying the product.

126. The method of claim 112 further comprising alkylating at least one amine in the product with an alkyl halide.

127. The method of claim 126, wherein the alkyl halide is methyl iodide.

128. 1A method of preparing a compound of claim 53, the method comprising steps of:
reacting a compound of formula:

wherein R1 is selected from the group consisting of hydrogen; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic;
cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic;

substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstitued, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; -ORA; -C(=O)R A; -CO2R A; -CN; -SCN; -SR A; -SOR A; -SO2R A; -NO2;
-N3;
-N(R A)2; -NHC(=O)R A; -NR A C(=O)N(R A)2; -OC(=O)OR A; -OC(=O)R A; -OC(=O)N(R A)2; -NR A C(=O)OR A; or -C(R A)3; wherein each occurrence of R A is independently a hydrogen; a protecting group; halogen; cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic; substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstitued, branched or unbranched aryl; substituted or unsubstituted, branched or unbranched heteroaryl; an acyl moiety;
alkoxy; aryloxy; alkylthio; arylthio; amino, alkylamino, dialkylamino, heteroaryloxy;
or heteroarylthio moiety; and R5 is independently selected from the group consisting of hydrogen and C1-C6 alkyl;
with an amine of formula:

wherein A is selected from the group consisting of cyclic or acyclic, substituted or unsubstituted, branched or unbranched aliphatic; cyclic or acyclic, substituted or unsubstituted, branched or unbranched heteroaliphatic; substituted or unsubstituted, branched or unbranched acyl; substituted or unsubstitued, branched or unbranched aryl; and substituted or unsubstituted, branched or unbranched heteroaryl;
to form a compound of formula:

129. A method of preparing microparticles, the method comprising steps of:
contacting a compound of claim 1 with an agent to be delivered in the presence of a solvent to form a mixture; and spray drying the mixture.

130. A method of administering an agent, the method comprising steps of:
administering to a subject in need thereof a therapeutically effective amount of a pharmaceutical composition comprising a compound of claim 1 and an agent to be delivered.

131. The method claim 130, wherein the agent is selected from the group consisting of polynucleotides, proteins, peptides, and small molecules.
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