WO1999033431A2 - Preparation and selection of pharmaceutically useful compounds from a diverse universal library - Google Patents

Abstract

This invention relates to novel combinatorial libraries of bridged biaromatic and triaromatic ring compounds, methods for preparing such libraries, and an apparatus for storing such libraries and for use as a component of assay systems for identifying compounds for drug development.

Description

METHODS FOR PREPARING AND SELECTING

PHARMACEUTICALLY USEFUL BRIDGED

BIAROMATIC AND TRIAROMATIC RING COMPOUNDS FROM

A STRUCTURALLY DIVERSE UNIVERSAL LIBRARY

Cross-reference to Related Applications

This application is related to copending Provisional Application Serial No. 60/070,021, filed December 30, 1997, incorporated by reference in its entirety, and claims the benefit of its filing date under 35 USC Section 119(e) .

Field of the Invention The invention relates to methods for preparing bridged biaromatic and triaromatic ring compounds having desired pharmaceutical or other biological utility. More particularly, the invention relates to structurally diverse libraries of bridged biaromatic and triaromatic ring compounds, methods for preparing such libraries, and an apparatus for storing such libraries and for use as a component of assay systems for identifying compounds for drug development .

Background of the Invention A key step in preparing and selecting pharmaceutically or other biologically useful compounds is identification of structurally-unique lead compounds. In 1990 it was estimated that nearly one- third of the $231 million average cost for making a new therapeutic compound available for widespread public use was spent in identifying and optimizing a lead chemical structure. Traditionally and currently mass screening of large numbers of compounds and mixtures of compounds has been and is the most successful method for identifying chemical leads. Recent availability of robotic, rapid, high throughput biological screens is beginning to make possible efficient screening of hundreds of thousands of compounds per year.

Most screening libraries consist of a historical collection of compounds synthesized in the course of pharmaceutical research, natural products, and, more recently, peptide libraries. Each of these libraries has limitations. Historical pharmaceutical collections of synthesized compounds contain a limited number of diverse structures which represent only a small fraction of total diversity possibilities. Limitations of natural products libraries include the structural complexity of the leads identified and the difficulty of reducing these leads to useful pharmaceutical agents. Peptide libraries are limited to peptides or peptide mimics. To date, conversion of peptide-chemical leads into pharmaceutically useful, orally active, non-peptide drug candidates in the absence of a small molecule chemical lead has been met with limited success.

Some of the peptide and peptide mimic libraries referred to above were prepared using combinatorial chemistry. A challenge facing medicinal chemists is to translate the success using combinatorial chemistry to prepare peptide and peptide- like compounds into technology suitable for efficiently preparing large libraries of low molecular weight non- peptide compounds. Solid phase chemistry for preparing low molecular weight compounds is desirable to effect such a translation. The following references are examples of the types of solid phase chemistry methods that may be useful in low molecular weight compound combinatorial chemistry. In 1974, F. Camps et al . (Annales De Quimica 70 , 848) reported solid phase synthesis of four related benzodiazepines . More recently Bunin and Ellman (J. Am. Chem. Soc. (1992) 114, 10997, and Proc. Nat. Acad. Sci. USA (1994) j31, 4708) and S.H. De itt et al .. (Proc. Nat. Acad. Sci. USA (1993) 90_., 6909) also reported preparation of a small number of benzodiazepines using solid phase chemistry. Two tetradecene-1-ol acetates also have been prepared on solid supports (CC. Leznoff et al . , Can. J. Chem.

(1977) 5_5, 1143) . Additionally, solid phase synthesis of 4 , 4 ' -stilbenecarbaldehyde has been reported (J.Y. Wong et al . , Angew. Chem. Int. Ed. (1974) 13., 666) . The following references are examples of biphenyl and triphenyl compounds that have been prepared by well known synthetic organic chemical methods. A.A. Patchett et al . recently reported that certain biphenyl acylsulfonamides and biphenyl sulfonylcarbamates are orally active antagonists of the angiotensin II receptor (Medicinal Chemistry Abstract #80 (1993) ACS Meeting-Chicago) . Other recently reported angiotensin II antagonists include several imidazopyridine and tetrazole-substituted biphenyl compounds (E.M. Naylor et al . , Medicinal Chemistry Abstract #76 (1993) ACS Meeting-Chicago) and a series of carbon-tethered biphenyl pyrrole compounds (J.M. Hamby et al . , Medicinal Chemistry Abstract #72 (1993) ACS Meeting-Chicago) . Others recently have reported that certain orthobiphenylphenols are leukotriene antagonists (M.J. Sofia et al . , Medicinal Chemistry Abstract #5 (1993) ACS Meeting-Chicago) .

Preparation of various other substituted biphenyls has been reported. An example of the many references describing methoxy substituted biphenyls is M.G. Banwell et al . which describes certain trimethoxy and tetramethoxy biphenyls that have tubulin binding properties (CA118 (19) :191308u (1992)). Another such reference describes synthesis of several methoxy and ethoxysubstituted biphenyls for use in a peroxidase indicator system for basis media (CA118 (1) :3411a (1992)). 2,4 ' , 5-Trimethoxy-4-biphenylcarboxylic acid has been reported to have estrogenic activity (CA54 :19584c (1959) ) .

Synthesis of 2 , 2 ' , 5, 5 ' - (tetrapropynl-1- oxy) biphenyl has been reported without indication of its use (CA116(11) :105745p (1991)). Similarly, 2 , 2 ' , 6 , 6 ' -tetrabenzyloxybiphenyl has been reported (CA110 (21) :192346b (1988)) and 2 , 2 ' , 3 , 3 ' - tetramethoxymethylbiphenyl (CA97 (11) : 91847y (1982)) have been reported without a suggested utility. Preparation of several trisubstituted and tetrasubstituted biphenyls and terphenyls has been reported (CA118 (21) :212566u (1993)).

Preparation of substituted bisphenyl compounds having a bridging group between the two phenyl rings has been reported. K. Edogawa et al. disclosed substituted bisphenyl compounds have S0₂, S, CMe₂, or 0 moieties between the rings that are useful in making semipermeable composite membranes for liquid separation (CA109 (18) : 151003y (1986)). Several tetrahydroxy substituted bisphenyl methanes without an indication of their utility have been reported (Marsh et al. Ind. Eng. Chem. (1949) 41, 2176) .

Thus, there remains a need for methods to efficiently prepare large libraries of low molecular weight non-peptide compounds and to select from such libraries compounds having desired pharmaceutical or other biological utility.

Summary of the Invention The present invention provides methods for preparing and selecting bridged biaromatic and triaromatic ring compounds having desired pharmaceutical or other biological utility, libraries prepared by such methods, and a system which utilizes such libraries for rapidly generating large rationally designed libraries of structurally diverse small molecule compounds to explore multiparameter space that overcomes many of the disadvantages associated with using currently available libraries as a basis for identifying and selecting new pharmaceutical agents. The present invention makes possible preparation of libraries of low molecular weight organic chemical compounds which have diverse chemical structures that are known and can be controlled. Additionally, other characteristics of the compounds that are important for pharmaceutical utility, such as solubility, can be controlled. Most importantly, however, because the compounds prepared using this invention are low molecular weight non-peptide compounds they are expected to be useful in a much broader spectrum of therapeutic applications than peptides which generally can only be administered by injection or inhalation. One embodiment of the present invention provides an assay kit for the identification of pharmaceutical lead bridged biaromatic and triaromatic ring compounds, said kit comprising assay materials and a well plate apparatus or equivalent apparatus providing a two-dimensional array of defined reservoirs. The well plate apparatus provides a diverse combinatorial library, wherein each well (reservoir) contains a unique member of the bridged biaromatic and triaromatic ring compound library. The well plate apparatus is used to provide multiple reaction zones for making the library and to store and provide a readily accessible source of library member compounds .

Detailed Description of the Invention

The method of the invention includes a multiple combinatorial approach to prepare structurally diverse libraries of biaromatic and triaromatic ring compounds which contain biologically useful compounds . Combinatorial chemistry takes advantage of the nature of the interaction between biological ligates such as antibodies, receptors, enzymes, ion channels, and transcription factors, and their ligands such as antigens, hormones, neurotransmitters, and pharmaceutical agents. It generally is agreed that ligate/ligand affinity and interaction results from binding or interaction between at least three functional groups or chemical functionalities on the ligand and complementary sites on the ligate. Strong interactions between ligates and ligands are dependent upon the properties and three dimensional spacial orientation of the functional groups or chemical functionalities on the ligands. High affinity specific ligands for a given ligate have functional groups that:

(1) bind tightly to the binding sites on the ligate and

(2) are positioned to bring the functional groups into close proximity with the ligate binding sites in the biological milieu where the interactions occur.

Compounds prepared according to present method have molecular weights of between about 200 and 1000 daltons, preferably between about 300 and 600 daltons, and comprise: (1) scaffold moieties and (2) at least two functional groups as components thereof. As used herein a "scaffold" is an invariable region or core of the compounds which are members of the combinatorial libraries of the present invention, onto which functional groups can be attached in a manner that when two or more scaffold moieties are attached results in the desired spacial orientation of the functional groups. Scaffold moieties preferably are selected such that they can be prepared from available materials by known chemical reactions and readily allow for attachment of desired functional groups and/or other scaffold moieties in a variety of positions on the molecule. In this specification and claims, as indicated by the context, "scaffold" may also refer to two or more attached scaffold moieties.

Suitable scaffolds are compounds of the following formula:

wherein:

M-. and M₂ independently are CH(NRR') or CH(OR) , wherein R and R' independently are H, C₀- C₆alkylCOR₁₅,

C₁.₆al ylOR₂₄, C₁.₆alkylNR_2SR₂₆, C₀__salkylNR₈₀C(NR₈₁)NR₈₂R₈₃, C^alkylindole, C₀_₆alkyl-D, C^ ₆alkylCN, C₀._εalkylSOR₁₅, C₀.₆alkylC (0) R₁₆, S00R₁₆, or C_x_

X₁₇ Y-. and Z_x are any accessible combination of CH, CHCH, 0, S, N, and NH provided that at least one is CH or CHCH and not more than one is CHCH;

X₂, Y₂ and Z₂ are any accessible combination of CH, CHCH, 0, S, N, and NH provided that at least one is CH or CHCH and not more than one is CHCH;

V is H,

halo, (C₀.₄alkyl) OH, (C₀_ alkylSH, (C₀_₄alkyl) NR₂₂R₂₃, (C₀.₄alkyl) C0₂R₇₆, or C₀_ ₄alkylOR₇;

W is H or

X₃, Y₃, and Z₃ are any accessible combination of CH, CHCH, 0, S, N, and NH provided that at least one is CH or CHCH and not more than one is CHCH. Preferred scaffolds are: (1) bisphenyl, (2) trisphenyl, (3) bispyridyl, and (4) pyridyl-phenyl wherein the bridging group between the rings is as defined for M_x and M₂ above. Useful functional groups include the side chains of the 19 naturally occurring L-amino acids and the side chains of nucleotides found in nature. Additionally, non-natural occurring mimics of these groups are useful. Preferred compounds of the invention which are prepared by combining preferred scaffold moieties with preferred functional groups are shown in Formula I below:

Formula I wherein:

X₁ , Y_x and Z are any accessible combination of CH, CHCH, 0, S, N, and NH provided that at least one is CH or CHCH and not more than one is CHCH; X₂, Y₂ and Z₂ are any accessible combination of CH, CHCH, 0, S, N, and NH provided that at least one is CH or CHCH and not more than one is CHCH; W is H or

X₃, Y₃ and Z₃ are any accessible combination of CH, CHCH, 0, S, N, and NH provided that at least one is CH or CHCH and not more than one is CHCH; _x and M₂ independently are CH(NRR') or CH(OR) , wherein R and R' independently are H, C₀-

C₆alkylCOR₁₅,

C₁.₆alkylOR₂₄, C₁.₆alkylNR₂₅R₂₆, C_0.6alkylNR₈₀C(NR₈₁)NR₈₂R₈₃, C^alkylindole, C₀_₆alkyl-D, C_x_ ₆alkylCN, C₀.₆alkylSOR₁₅, C₀.₆alkylC (0) R₁₆, S00R₁₆, or C_x. _galkylOC^_galkylR^; V is H, C^alkyl, halo, (C₀.₄alkyl) OH, (C₀.

₄)alkylSH, (C₀.₄alkyl) NR₂₂R₂₃, (C₀.₄alkyl) C0₂R₇₆, or C₀. ₄alkylOR₇ ;

A_χ/ A₂, A₃, A₄, A₅, and A₆ independently are absent or present as 0, S, NR₆₀; or C₀.₆alkylC (0) NR₂₁, provided that at least two are present ;

R₁₇ R₂, R₃, R₄, R₅, and R_s independently are H, C_0.6alkylCOR₁₅, C₁.₆alkylR_1gR₁₇, C₁.₆alkylOR₂₄ except methoxymethyl ,

C₀_₆alkylNR₈₀C (NR₈₁) NR₈₂R₈₃, C^_galkylindole, C₀.₆alkyl-D, C^alkylCN, C₀.₆alkylSOR₁₅, or C^_galkylOC^_galkylR^;

D is one or multiple fused saturated or unsaturated five or six membered cyclic hydrocarbon or heterocyclic ring system containing one or two 0, N, or S atoms that is unsubstituted or substituted by an accessible combination of 1 to 4 substituents selected from C^_galkyl, NR₇R₈, 0R₉, SR₁₀, COR^, halogen, CF₃, or 0;

^•K-7 ' ^8' ^9 ' " 0 ' "19' "20' 21 ' "-22' "23' "60' "80'

R₈₁, R₈₂, and R₈₃ independently are H or

"12 ' "13' "-14 ' "16' "17' "18 ' "24' "25' "26' and R_?g independently are H, C₁.₆alkyl, phenyl, or substituted phenyl ;

R_1X is 0R₁₂ or NR₁₃R₁₄; R₁₅ is 0R₁₈ or NR₁₉R₂₀; or any pharmaceutically useful salt thereof. The compounds of Formula I constitute a universal library of compounds that includes pharmaceutically useful compounds. As used in Formula I and elsewhere in this specification and the claims, "C_x._yalkyl" is a straight chain or branched, saturated or unsaturated alkyl group containing x to y carbons atoms wherein x and y are integers and "halo" includes bromo, chloro, fluoro, and iodo, and "substituted phenyl" is a phenyl group substituted by any accessible combination of halo, CF₃, OH, C^alkyl, C^alkoxy, COOH, COOC₁_₆alkyl , NRR', CONRR', CN, O, N0₂, C₀.₆alkylCOR_ls, or C₀.₆alkylC (O) R₁₆, wherein R and R' independently are H or C_halky1.

In more preferred compounds of the invention X_x to X₃, Y-. to Y₃, and Z_λ to Z₃ are selected so that one or more of the ring systems is pyrrole, furan, thiophene, pyridine, pyrazole, pyrimidine or isoxazole with phenyl being most preferred. Also in more preferred compounds of the invention D is one of the following ring systems substituted as described above: pyrrole, furan, imidazole, thiophene, pyridine, pyrazole, pyrimidine, pyridazine, or isoxazole with phenyl being most preferred.

More preferred compounds of the invention are shown in the following Formula II:

Formula II wherein:

Mi is CH(O(C₀.₄alkyl)R) , wherein R is H, C_x_ ₆alkyl, D, COOH, COOC^alkyl , OH, OC^alkyl, NH₂, N(C_X. ₆alkyl)₂, or CN;

V_x is H, CH₃, OH, or CH₂OH; A₇, A₈, A₉, and A₁₀ independently are absent or present as 0 provided that three are O; and R₃₀, R₃₁, R₃₂, and R₃₃ independently are H, C _ ₆alkyl, D, COOH, COOC^alkyl, OH, OC^alkyl, NH₂, N(C₁. ₆alkyl)₂, or CN.

Pharmaceutically useful salts of the above compounds include, for example, sodium, potassium, trialkyl ammonium, calcium, zinc, lithium, magnesium, aluminum, diethanolamine, ethylenediamine, megulmine, acetate, maleate, fumarate, lactate, oxalate, methansulfonate, ethanesulfonate, benzenesulfonate, tartrate, citrate, hydrochloride, hydrobromide , sulfate, phosphate, and nitrate. Other pharmaceutically useful salts are readily apparent to skilled medicinal chemists.

Some of the compounds included in Formula I can exist in more than one chiral form and thus exhibit stereoisomerism. Formula I includes all purified stereoisomers and racemic mixtures of the compounds within its scope.

In one aspect of the invention a preliminary step in preparing and selecting compounds having desired pharmaceutical or other biologic utility is preparation of a universal library. As stated above medicinal chemists and pharmacologists generally agree that interactions between biological ligates and ligands require that the ligand contain at least three functional groups in a spacial orientation that is complementary to the binding sites on the ligate. It also is known that the distance between the binding sites on ligates is determined by the conformation of the ligate as it exists in its native environment and that effective ligands are those that have functional groups positioned to be complementary to such conformation. Because ligates are three dimensional in their natural setting, for any selected intramolecular distance between binding sites an essentially infinite number of possible specific positions for the binding sites exist . Thus there similarly is a very large number of possible functional group positions on the ligands that effectively interact with particular ligates. As used in this specification and claims a universal library is a collection of related small molecular weight compounds that with respect to spacial orientation of functional groups effectively samples a large segment of the possible specific positions with a selected distance and a sub-universal library is a universal library that is targeted to a particular biological ligate. The number of compounds in the library can vary. Preferably the library includes at least ten compounds, more preferably at least about 100 compounds , or more .

Preparation of bradykinin antagonists provides an example of the general approach to designing a sub-universal library. Bradykinin is a naturally-occurring nonapeptide (Arg-Pro-Pro-Gly-Phe- Ser-Pro-Phe-Arg) that is formed enzymatically in the blood and extracellular fluids after injury (a review covering all aspects of bradykinin has appeared (M. Hall, Pharmac. Ther. 56_. (1992) 131) .

At least two distinct receptor types, Bl and B2 appear to exist. Although activation of B12 receptors appears to underlie the most relevant biological actions of kinins, both Bl and B2 receptors could be important in developing therapeutic strategies. Bradykinin is a major pain producing substance that excites and sensitizes sensory nerves following trauma, burns, injury and infection. Peptide bradykinin antagonists block bradykinin-induced pain in animal models suggesting that a bradykinin antagonist would be effective for the treatment of a variety of painful disorders. Bradykinin has also been found in plasma exudates taken from the scalp of migraneurs and has been shown to cause severe vascular head pain upon intravenous injection suggesting that bradykinin antagonists would be useful for the treatment of headache . Bradykinin is a potent vasodilator of most peripheral arteries and also causes neurogenic inflammation by the peripheral release of substance P, neurokinin A, and CGRP from sensory nerve fibers. Bradykinin has also been found in fluid from arthritic joints. These results suggest that bradykinin antagonists might have an important role as antiinflammatory agents. Bradykinin has been proposed to play a role in the pathogenesis of asthma as well. While an orally-active bradykinin antagonist is likely to be of immense therapeutic benefit, the potent bradykinin agonists and antagonists reported to date have been peptide derivatives similar in size to bradykinin (which like bradykinin are expected to be rapidly degraded in body fluids) .

Peptide analogs of bradykininhave shown that in general, replacement of Pro 7 with D-He or conformationally-constrained analogues as well as replacement of the Phe 5 and 8 with thienylalanine or conformationally-constrained phenyl analogues affords competitive and selective antagonists of bradykinin. The C-terminal arginine is crucial for receptor activity. It appears that the N-terminal amino groups is not necessary for activity since it can be acylated or removed without significant loss of activity. Bl selective antagonists are obtained by making the des-9 Arg analogues .

As an example, D-Arg°-Hyp³-Thi⁵-D-Tic⁷-Oic⁸- bradykinin is a specific, potent, and long-lasting bradykinin antagonist being developed by Hoechst (Hoe- 140) for allergic rhinitis and asthma. Furthermore, Kyle et_. al . have incorporated unnatural amino acids in the C-terminus of bradykinin which introduce B-turn stability and conclude that a B-turn in the four C- terminal amino acid residues might be a prerequisite for high receptor affinity (D.J. Kyle et_. al . , J. Med. Chem (1991) 3_4 (3) : 1230-33) . Using the information described above it is possible to design a sub-universal library that is likely to possess bradykinin antagonist activity. The B-turn likely at the C-terminal portion of bradykinin suggests that the peptide antagonists are not fully extended at the receptor and likely occupy a distance of 10-18A. This is an ideal size to be mimicked by a bisphenyl scaffold and the size, shape, and group variations are explored by preparing a large library of compounds guided, or limited, by previously reports SAR studies on bradykinin receptor antagonists. This approach can be carried over to B₁ receptors by leaving out the arginine mimic on the A-ring. Using the previously described SAR data on bradykinin peptide antagonists the following compounds of Formula III are expected to include bradykinin antagonists:

Formula III wherein:

M_j. and M₂ independently are CH(NRR') or CH(OR) , wherein R and R" independently are H, C₀- C₆alkylCOR₁₅,

C₁.₆alkylNR₂₅R_2fi, C_0.6alkylNR₈₀C(NR₈₁)NR₈₂R₈₃, C₀.₆alkyl-D, C^alkylCN, C₀_ ₆alkylSOR₁₅, C₀.₆alkylC (0) R₁₆, SOOR₁₆, or

₆alkylR₁₅;

B and B ' are H, O (CH₂) _nNR₄₀C (NR₄₁) NR₄₂R₆₁, or O (CH₂)_n,NR₄₃R₄₄ wherein R₄₀, R₄₁, R₄₂, R₄₃, R₄₄, and R₆₁, independently are H or C^alkyl, n and n' are 2 or 3; provided one of B and B' is H; E is

wherein X is CH, N, NH, 0, S; n is 1-3; n' is 1 when X is 0,S, or NH; and n' is 2 when X is CH or N;

F, F', and F" are H, 0 (CH₂) _nNR₄₅C (NR₄₆) NR₄₇R₆₂, or 0(CH₂)_n,NR₄₈R₄₉ wherein R_4S, R₄₆, R₄₇, R₄₈, R₄₉, and R₆₂ independently are H or C^alkyl, and n and n' are 2 or 3; provided two of F, F ' , and F" are H;

G and G' are H, O(CH₂)_nOR₅₀, or

wherein X' is CH, N, NH, 0, or S'; R₅₀ is H or C^alkyl; and

R₅₁ is H, C₁.₃alkyl, halogen, OH, or OC-^alkyl ; n is 1-3; and n¹ is 1 or 2; provided one of G and G' is

H.

In preferred compounds:

B or B' is OCH₂CH₂NHC(NH)NH₂;

E is

or

F¹ and F" are OCH₂CH₂NHC (NH) NH₂; and G and G' are H.

These compounds are tested for bradykinin antagonist activity using a high-volume biochemical binding assay such as is referenced in the examples below. For potential use in rapid mass screening, a rat B2 receptor has been cloned by Jarnagin et. al . IPNAS (1991) ' 7724) . It appears to be a 7- transmembrane domain G-protein coupled receptor with a molecular weight of 42 kD and 366 amino acids. Furthermore, human B2 receptor was cloned by Hess et al. (Biochem. Biophys. Res. Comm. (1992) 184, 260) and has a molecular weight of 41.1 kD and 364 amino acids with 81% sequence homology to the rat B2 receptor. The binding assays are followed by examination of the compound in an in vitro smooth muscle preparation. Functional activity is assessed by examining in vitro PI turnover. In vivo models include bradykinin paw pressure in rate, both IP and PO.

Most of the remaining seven transmembrane G- protein coupled receptors (GPCR) are viable candidates for the approach described herein. Such receptors include, but are not limited to, CCK, angiotensin, bombesin, bradykinin, endothelin, neuropeptide Y, neurotensin, opiod, somatostatin, tachykinin (NK-1, NK- 2, NK-3) , thromboxane A₂, and vasopressin. The angiotensin-2 receptor might be of particular interest as a test case in light of the recently reported activity of a number of functionalized bisphenyl molecules.

The ligands for many of the GPCRs range from small-medium sized organics to small-medium peptides (4-35 amino acids) . Most of these ligands are expected to occupy a 10-30 cubic A volume making them ideal candidates for the libraries described herein. An increasing number of modeling and mutagenesis studies are not only indicating the appropriate approximate size but are also giving specific information on important residues of the receptor that interact with the ligand. This information can be readily applied to the design of receptor specific sub-universal libraries .

Some examples of recently available information includes the TXA₂ receptor (Yamamoto et, al . , J. Med. Chem (1993) 3_6, 820-25) . These workers propose the TXA₂ binding site and suggest specific residues of the receptor that are important for ligand binding, including Ser-201, Arg-295, and Trp-258. Groups that are complimentary to these residues would be built into the sub-universal library. The NK 1-3 receptors have been cloned and expressed and mutational studies are ongoing which suggest the binding site for NK-1 antagonists is likely to be around the junction of extracellular loop 2 and the top of TMV and TMV1. Furthermore, the identification of non-peptide leads for the NK-1 receptor suggests some groups that allow initial selection of groups for a sub-universal library (Watling, TIPS (1993) 14, 81) . It is believed that NK- 1 antagonists will be useful for treating pain, inflammation, arthritis, and asthma.

Identification of residues for design of a somatostatin sub-universal library is guided by the work of Hirschmann et. al . (J. Amer. Chem. Soc. (1992) 114, 9217) . Preparation of compounds that interact with ion channels provides another example of designing a sub-universal library. Ion channels are proteins which span cell membranes providing pathways for the flow of ions such as chloride or potassium. These channel proteins are involved in many cellular functions such as nerve signaling, muscle contraction and hormone secretion. Over the past several years there has been an explosive growth in the number of cloned and expressed ion channels, as well as in discoveries which link channels to disease. Moreover, now that it is clear that there are many subtypes of ion channels, differentially distributed throughout the body, the possibilities for selective targeting of specific channels in specific tissues are unlimited. This selective targeting will reduce unwanted drug-related side effects and toxicities. Potassium channels can be divided into at least 6 major classes, and 15 subclasses, each with its own distinct biophysical and pharmacological identity. Agents which modulate specific potassium channels in specific tissues are expected to target select disease states withoutaltering normal functions. Potassium channels are largely responsible for maintenance functions like establishing the membrane potential in unstimulated cells, or in switching on, or off, a cell's electrical activity. Thus, these channels in part control the cell ' s capacity for nervous transmissions, muscle contraction and secretion. Due to their integral roles in almost all normal signal processing, agents which modulate potassium channels are likely to be useful for treating conditions such as diabetes and muscular sclerosis, cardiac arrhythmias and vascular hyperactivity.

Various types of ligand-activated and voltage-activated ion channels have now been cloned and functionally expressed. Sequence comparisons and hydropathy analyses have revealed a great deal of structural homology among these channels. Each channel sequence is composed of a repeating motif of transmembrane spanning domains which combine in various ways to form channels (For a recent review of the field, see Andersen and Koeppe, II, Physiological Reviews (1992) Vol. 72) . Site-directed mutagenesis has allowed researchers to probe the primary structure of ion channel proteins for critical amino acid residues involved in the binding sites of drug molecules. These studies will allow for the development of agents targeted for specific channel subtypes and binding sites. To date, several classes of ion channels, including potassium and chloride, have received intensive characterization leading to a basis on which to consider structure-based drug design.

Toxins, such as those from scorpion venoms, have proven useful in defining potential drug interaction sites on ion channels as well as defining physiological roles for channels. These peptide toxins which are 36-38 residues long, contain three disulfide bridges, and share strong sequence similarity among isoforms, block both voltage-gated and Ca-activated K channels with nanomolar affinity. Within this group of toxins, ther are specific subtypes which bind to specific subtypes of potassium channels. Electrostatic interactions between charybdotoxin (CTX, a specific peptide pore blocker of K channels and a Ca-activated K channel have been extensively investigated. Charybdotoxin has eight positively charged residues (four lysines, three arginines, and one histidine) .

Electrostatic forces are known to favor CTX binding to the negatively charged mouths of K channels. However, only replacement of Arg25, Lys27, or Lys34 with a Gin residue strongly decreased the affinity of the toxin for the channel. These three residues are located close to one another on one side of the CTX molecule and make direct contact with the channel mouth. On the opposite side are five charged residues whose neutralization show little effect. Therefore the positively charged groups on CTX promote toxin channel interaction in two ways; by weak electrostatic influences and by direct and intimate contact with the channel on one side of the toxin molecule. The solution structure of CTX has been recently determined (Bontems et al ■ , Biochemistry (1992) 3_1, 7756) and it has been shown that Arg 25 and Lys34 are located within A of Lys 37 and each is crucial for high affinity binding of CTX. The receptor site in the channel's mouth must be wide (>22A) and flat to accommodate the CTX molecule. The wide mouth must narrow abruptly into an ion-selective pore in order to provide a selective K binding site with which lys27 interacts (Miller and

Park, Biochemistry (1992) 3_1, 749, and Neuron (1992) 9_., 307) . These studies reveal a molecular surface of CTX which make direct contact with the extracellular mouth of the K channel and a single CTX molecule physically occludes the K conduction pathway by binding to a receptor located in the externally-facing mouth of the channel protein.

Using the information described above, a sub- universal library targeted to K channels which mimics the three important binding residues both electronically (three positive charges) and spatially (6-18A total spearation) is designed. Such a library is expected to identify non-peptide CTX mimics with therapeutic potential. The compounds of Formula IV represent a sub-universal library targeted to potassium channels :

Formula IV wherein: M_x and M₂ independently are CH(NRR') or CH(OR), wherein R and R' independently are H, C₀- C₆alkylCOR₁₅,

C₀.₆alkylNR₈₀C(NR₈₁)NR₈₂R₈₃, C^alkylindole, C₀._salkyl-D, C_x. ₆alkylCN, C₀.₆alkylSOR₁₅, C₀.₆alkylC (0) R₁₆, S00R₁₆, or C_x.

J, J', and M independently are O(CH₂)_nNR₅₀C(NR_sl)NR₅₂R₆₅ or 0 (CH₂) _n,NR₅₃R₅₄ wherein R₅₀, R₅₁, ^R ₅₂' ^R ₅₃' ^R ₅₄' ^R ₆₅ independently are H or

and n and n' independently are 2-3;

Q and Q' are H or 0 (C₁.₄alkyl) T wherein T is C^alkyl, C0₂R₅₅, OR₅₆, or

wherein:

X₇ is CH, N, NH, S, or 0; n* ' ' is 1 or 2;

U is H, C^_galkyl, halogen, CF₃, or OR₅₇; and R_55/ R₅₆, and R₅₇ independently are H or C_x.

₆alkyl; provided that at least Q or Q' is H.

The presently invented multiple combinatorial method for preparing and selecting small molecular weight compounds having pharmaceutical utility or other biologic utility is used to efficiently prepare universal libraries. As used herein a multiple combinatorial method is a method for preparing compounds that uses two or more scaffold molecules each carrying functional group (s) that have been attached in a combinatorial fashion. Generally, compounds comprising two scaffold moieties are used for ligates of about 12 to 20A and compounds having three scaffold moieties yield ligands for ligates of about 20 to 35A. The power of the invented multiple combinatorial method is demonstrated by the numbers of compounds that can be prepared quickly and efficiently. Libraries with greater number of compounds can be readily prepared in accordance with the invention. For example, using two scaffold molecules each containing two of twenty possible functional groups arranged in four different orientations yields more than 1,000,000 compounds. Using the same parameters with a third scaffold molecule allows for preparation of a universal library containing more than 1,000,000,000 compounds. The compounds of Formula I are an example of a universal library of compounds that are prepared according to the invention. In another aspect, the invention is used to prepare large quantities of a desired target compound rather than small amounts of multiple compounds as is the case in preparing universal or sub-universal libraries.

Preferably when preparing universal or sub- universal libraries, multiple compounds are prepared by simultaneously conducting different chemical reactions in multiple reaction vessels, as discussed below. To prepare large quantities of a selected compound the same reaction is carried out simultaneously in different vessels.

The process of the invention may be carried out in any vessel capable of holding the reaction medium. In one embodiment, the process of the invention is carried out in containers adaptable to parallel array synthesis. The process can be conducted in an apparatus providing multiple reaction zones, typically a two-dimensional array of defined reservoirs, wherein one member of the libraries of the invention is prepared in each reservoir. Thus the library of the invention comprises a plurality of reservoir arrays, e.g., well plates or equivalent apparatus, each reservoir or well containing a unique member of the library. Accordingly the library compounds are typically identified by reference to their well plate number and their x column and y row well plate coordinates. Following simultaneous preparation of the library member compounds in the reservoir array, the compounds can be transferred in whole or in part to other reservoir arrays to prepare multiple copies of the library apparatus or to subject the library to additional reaction conditions. Copies of the library apparatus (each comprising a two-dimensional array of defined reservoirs with each reservoir containing a predetermined member of the library) are useful as replaceable elements in automated assay instruments. The apparatus of the invention allows convenient access to a wide variety of structurally related compounds. One preferred reservoir array for use in making and using this invention is a multi-well titer plate, typically a 96 -well microtiter plate. The compounds and libraries of the invention preferably are prepared according to Scheme I below. In Scheme I the preferred method of synthesizing the compounds on a solid support is depicted. The libraries and compounds of the invention, however, also can be prepared using solution phase chemistry.

[This space intentionally left blank]

Scheme I

(1) (2)

1. Deprotect

2. Introduce Functional

Group *~ er

3. Repeat if Desired

4. Activate X' for Gh Link

Coupling (3)

1. Deprotect

2. Introduce Functional

Group *^*-

3. Repeat if Desired

(5)

1. Remove Protecting Group

2. Cleave from Solid Support

(6)

(7) Scheme I demonstrates the invented method of preparing universal libraries of compounds. According to this scheme functional groups are attached to a first scaffold moiety to yield a compound comprising a scaffold and one or two functional groups (Compound 3) . Thereafter a second scaffold molecule (Compound 4) is added with the formation of M followed by addition of functional group (s) to the second scaffold moiety to yield Compound 6 which can have three or four functional groups. Compounds of Formula I then are prepared by cleaving Compound 6 from the solid support . Compounds of the invention wherein M_x is -CH(OR)- are prepared as described in Example 1.

In Scheme I "SS" is a solid support material such as the cross-linked polystyrene resin known as the Merrifield resin (R.S. Merrifield, J. Am. Chem. Soc. (1963) 15_., 2149) . Alternatively, any other suitable polymeric resin or other support material such as, for example, silica, glass, cotton, and cellulose is used. Also in Scheme I "AG" is any suitable group for attachment to the linker such as, for example, OH, NH₂, COOH, CH₂OH, CH₂Br, CHO, CH₂Cl, CH₂SH, SH and V is the same as in Formula I .

The linker group shown in Scheme I is any group that hold the first scaffold (Compound I) onto the solid support and is formed by reaction of AG with the solid support, is stable to the reaction conditions necessary to complete the synthesis, and is easily cleavable upon completion of the synthesis. Suitable linkers are, for example, an OH, NH₂, halogen, SH or COOH group. An olefin group also is used as a linker. In such case, for example, AG in Compound 1 is CHO and it is attached to the solid support using a Wittig-like reaction. When an olefin group is used the final product is cleaved from the linker by treatment with ozone or other known methods. A sulfide or oxygen bond is another suitable linker. When a sulfide or oxygen bond is the desired linker AG in Compound 1 is CH₂halogen, preferably CH₂Br, and the bond between the solid support and Compound 1 is formed by reaction between the AG on Compound 1 and an SH or OH group on the solid support. Upon completion of the synthesis a sulfide or oxygen bond linker is cleaved by, for example, treatment with hydrogenolysis, Raney nickel, or dissolving metal reduction.

When a benzyl ester group is the desired linker AG in Compound 1 is CH₂OH and the bond between the solid support and Compound 1 is formed by reaction between the AG on Compound 1 and a C0₂H group on the solid support . Upon completion of the synthesis the benzyl ester group is cleaved by, for example, hydrogenolysis conditions.

P and P' in Scheme I are protecting groups for aromatic hydroxy groups. P and P' can be the same or different to allow for selective deprotection. Choice P and P' also is influenced by compatibility with the chemistry to be used in the remainder of the synthesis. Preferred protecting groups are C(0)CH₃ and Ph-Co wherein "Ph" is phenyl. Deprotection of a C(0)CH₃ is performed by treatment with an amine according to known procedures and deprotection of a Ph-CO group is accomplished by treatment with a nucleophile such as methoxide using known conditions and procedures.

In Scheme I X' and Y' are groups that allow for formation of M_r A preferred method for joining the rings is through reaction of a trimethylstannyl group on Compound 1 wherein X' is SnMe₃, with an acid chloride on Compound 4 wherein Y' is C0₂C1. Alternatively, a preferred method for joining the rings is through reaction of an OH group on Compound 1 wherein X' is OH with an acid chloride, I, or CH₂Br. When compounds having more than two scaffold moieties are desired the procedure of Scheme I is modified by repeating the steps needed to add one or more additional scaffolds before cleaving from the solid support. Also, the general procedure shown in Scheme I is used when scaffolds other than phenyl rings are used. Thus, any of the compounds included in Formula I can be prepared using Scheme I modified as may be necessary to accommodate different scaffold moieties. Any such necessary modifications are apparent to those skilled in the organic chemical synthetic arts. As used in Scheme I "FG" is a functional group which may be the same or different at different positions on the compounds. Suitable functional groups are the R through R₆ groups as defined in Formula I above. Although Scheme I shows preparation of compounds having two scaffold moieties and four functional groups such compounds having three functional groups are prepared by using a scaffold having one functional group in place of Compound 1 or Compound 4. Also, Compounds 1 and 4 provide for attachment of functional groups through an oxygen. By suitable replacement of these compounds a sulfur atom, a nitrogen atom, or an N-alkylamide group can be used in place of one or more of the oxygens . Procedures for introducing functional groups onto scaffolds are included in the examples below.

Scheme II is a modification of Scheme I procedure that is used to prepare compounds wherein the functional group is attached to the scaffold moiety using a (CH₂)_n,C (0) NR' and n' is O and R' is H or C_x_ ₆alkyl . In Scheme II AG, X', Y', and FG have the same meanings as in Scheme I . Scheme II

( ssj— Urker +

(8) (9)

1. Hydrolyze

2. Introduce

Functional QOu s

(13) According to Scheme II a scaffold molecule having two cyano groups attached (Compound 8) first is attached to a solid support via a linker and then is hydrolyzed to yield free carboxylic acid groups (Compound 10) . Then, functional groups are attached by treatment with HN(CH₃)FG to yield a scaffold with two functional groups (Compound 11) . Next a second scaffold moiety with two cyano groups is attached as described in Scheme I followed by addition of functional groups to yield Compound 13. Compounds to be included in the libraries of the invention then are prepared by adjusting the M_x group as needed, deprotecting and cleaving Compound 13 from the solid support as described in Scheme I . Scheme III describes an alternate method of producing compounds wherein the functional groups are linked to the scaffold moieties via a C(0)N(CH₃) residue. In Scheme III X', Y', P, P', and FG have the same meanings as in Scheme I .

[This space intentionally left blank]

Scheme I I I

1. Deprotect

2. Introduce Functional Groups

(14) 3. Repeat if

Desired (15)

1. Conversion of X, if needed

(17) (16) *

1. Deprotect

2. Introduce Functional Groups

3. Repeat if Desired

(18)

According to Scheme III Compound 14 is prepared by adding HN(CH₃)P or HN(CH₃)P' to the COOH functionalities of Compound 10 from Scheme II. Compound 15 is then prepared by deprotecting, differentially if desired, and introducing functional groups onto Compound 14. Compound 16 then is added to Compound 15 using the procedure for attaching scaffold moieties described in Scheme I to yield Compound 17. Compound 18 next is prepared by deprotecting, differentially if desired, and introducing functional groups onto Compound 17. Compounds included in the invented libraries are prepared by adjusting the M group as needed, and deprotecting and cleaving Compound 18 from the solid support as described in Scheme I. Scheme IV describes an alternate method of producing compounds wherein the functional groups are linked to the scaffold moieties via a C(0)N(CH₃) residue. In Scheme IV, X', Y', P, P' and FG have the same meanings as in Scheme I .

[This space intentionally left blank]

Scheme IV

1. Deprotect

2. Introduce Functional Groups

(19) 3. Repeat if

Desired (20)

(22)

(21)

1. Deprotect

2. Introduce Functional Groups

3. Repeat if Desired

(23)

The starting compound in Scheme IV (Compound 19) is prepared by standard procedures. Compounds included in the invented libraries are prepared by cleaving Compound 23 from the solid support. Alternatively, two or more scaffolds are independently derivatized with one or two functional groups, then are combined in a convergent approach. In a preferred method of this approach, two scaffolds are independently attached through a separate linker to a separate solid support material. The linkers and solid supports can be the same or different . The scaffolds can have handles for introducing side chains that are optionally protected or differentiated as described herein. After the attachment of one or two functional groups to each scaffold, one derivatized scaffold can be cleaved from its solid support, then reattached to the other scaffold through an appropriate coupling reaction. After any additional desired or needed synthetic transformations, (e.g., side chain protecting group removal), the functionalized scaffold (s) is cleaved from the remaining solid support to give compounds of the invented libraries.

Alternatively, two scaffolds can be prepared independently with desired functional groups attached thereto. The functionalized scaffolds can then be attached to each other through an appropriate coupling reaction, and the coupled scaffolds attached to a solid support material and manipulated as desired (e.g., side chain protecting group removal, derivatization of linking group of the scaffolds) . The resultant scaffold can then be cleaved from the solid support to give compounds of the libraries of the invention.

When compounds having more than two scaffolds are desired, a third scaffold can be independently functionalized, then coupled in the desired manner to one or both of the other scaffolds attached to a solid support or a combination of the two strategies can be employed whereby two scaffolds are attached together on a solid support in the manner described in the Schemes herein (a linear approach) , then a third functionalized scaffold derived from a separate solid support is attached. In any case two or more scaffolds can be separately functionalized in a parallel, simultaneous fashion.

The disclosed invention includes the following Formula V compounds which are useful as intermediates in preparing the invented libraries and compounds :

Formula V wherein :

W is H or

X₃, Y₃, Z₃, A₅, and A₆ are as defined in Formula I ;

R'_lf R'j, R'₃, R'₄, R'₅, and R'₆ are a protecting group or R_ιr R₂, R₃, R₄, R₅, and R_s as defined in Formula I, provided that at least one R ¹ ₁ to R'_ε is a protecting group;

V is V as defined in Formula I or a bond to a solid support; and the remaining variables are as defined in Formula I . As used in Formula V, a protecting group is any of the well known protecting groups that is suitable in view of the synthetic conditions used. Preferred protecting groups are C(0)CH₃ and Ph-CO. Preparation of libraries of Formula I compounds is the first step in the invented method of preparing and selecting compounds having pharmaceutical or other biologic utility.

In one embodiment of the present invention, an assay kit is provided for the identification of pharmaceutical lead compounds. The assay kit comprises, as essential parts, (1) a well plate apparatus (containing one of the compounds of the present invention in each of its individual wells) , and (2) biological assay materials. The biological assay materials are generally known to be predictive of success for an associated disease state. Illustrative of biological assay materials useful in the kit of this invention are those required to conduct assays known in the art, which include, but are not limited to: in vitro assays such as enzymatic inhibition, receptor- ligand binding, protein-protein interaction, protein- DNA interaction, and the like; cell-based, functional assays such as transcriptional regulation, signal transduction/second messenger, viral infectivity, and the like; and add, incubate & read assays such as scintillation proximity assays, fluorescence polarization assays, fluorescence correlation spectroscopy, colorimeric biosensors, receptor gene constructs for cell based assays, cellular reporter assays utilizing, for example, reporters such as luciferase, green fluorescent protein, B-lactamase, and the like; electrical cell impedance sensor assays; and the like. Generally, any activity identified in vitro is confirmed by evaluation in a suitable animal model if such is available and predictive of human pharmaceutical activity. The examples below include assays that are useful to select compounds of the invention that have pharmaceutical utility.

The compounds of Formula I that are useful as pharmaceutical agents can be incorporated into convenient dosage unit forms such as capsules, tablets, or injectable preparations. Pharmaceutical carriers which can be employed include, among others, syrup, peanut oil, olive oil, and water. Similarly, the carrier or diluent may include any time delay material, such as glycerol monostearate or glycerol distearate, alone or with a wax. The amount of solid carrier will very widely but, preferably, will be from about 25 mg to about 1 g per dosage unit. If a liquid carrier is used, the preparation will be in the form of a syrup, emulsion, soft gelatin, capsule, sterile injectable liquid such as an ampule, or an aqueous or non-aqueous suspension.

Pharmaceutical preparations are made following conventional techniques of a pharmaceutical chemist involving mixing, granulating, and compressing, when necessary, for tablet forms, or mixing, filling, and dissolving the ingredients, as appropriate, to give the desired oral or parenteral end products. Doses of the pharmaceutically useful compounds of the invention will be an effective amount, that is, an amount necessary to produce the desired effect without producing untoward toxicity selected from the range of 0.1-1000 mg/kg of active compound, preferably 10-100 mg/kg. The selected dose is administered to a patient in need of treatment from 1-5 times per day, orally, rectally, by bolus injection, or by infusion.

The present invention will be further illustrated by the following non-limiting examples. EXA P E 1

General Procedures for Formula I Wherein M_x is -CH(OR)-

A-Rincr Synthesis

5-Iodo-2-hydroxybenzaldehvde (2)

2-Hydroxybenzaldehyde (42 mL, 400 mmol) was dissolved in 400 mL of 1.0 M ICl/CH₂Cl₂ solution under argon and stirred for 12 hours. The reaction mixture was concentrated and redissolved in CH₂C1₂ three times and washed with saturated sodium bisulfite solution. The organic layer was dried with MgS0₄ and once again evaporated to dryness. The crude product was recrystallized from CH₂C1₂ to give 30.6 g (124 mmol, 31% yield) of pure 2..

Acetal Formation (3) 2-Hydroxy-5-iodobenzaldehyde (28.50 g, 115 mmol) was combined with excess 1, 3-dihydroxypropane (42 mL, 575 mmol) in dry toluene (150 mL) under argon in a 250 mL round bottomed flask equipped with a Dean-Stark trap. A catalytic amount of p-toluenesulfonic acid

(0.1 g) was added and the solution refluxed for 10 hours. The mixture was cooled, washed with two 100 mL portions of water, dried over MgS0₄, and evaporated to dryness . The crude product was purified by column chromatography over silica gel to give 33.6 g (109 mmol, 95% yield) of the acetal .

Alkylation of the Phenol

The acetal (3_) (5.03 g, 16.4 mmol) was dissolved in dry THF (50 mL) under argon. Phenethyl alcohol (2.0 mL, 16.4 mmol) and triphenylphosphine (4.30 g, 16.4 mmol) were added and the solution cooled to 0°C. Diisopropyl azodicarboxylate (3.3 mL, 16.4 mmol) was added dropwise with stirring. The solution wa allowed to come to room temperature over 10 hours. The reaction mixture was diluted with Et₂0 (150 mL) , washed with 3 x 100 mL portions of water and dried over MgS0₄. After concentrating, the crude product was purified by column chromatography to give 5.96 g (89% yield) of 4_..

Deprotection of the Acetal

The acetal (4.) (5.96 g, 14.5 mmol) was dissolved in 50 mL of THF prior to slow addition of 40 mL deionized water and 15 mL of cone. HCl. The mixture was vigorously stirred for 8 hours, extracted with 100 mL of Et₂0 and washed with 3 x 100 mL portions of deionized water. The organic layer was dried with MgS0₄ and concentrated and the crude product purified by column chromatography to give 4.57 g (90% yield) of the aldehyde 5_.

Stannylation of the A-Ring

The iodo compound 6. (4.57 g, 13 mmol) was dissolved in 50 mL of toluene under argon. Hexamethyl ditin (2.8 mL, 13 mmol) and the tetrakis (triphenylphosphine) palladium (0) catalyst (0.72 g, 0.6 mmol) were added and the mixture brought to reflux. After 20 minutes a palladium mirror formed on the inside of the flask and the reaction was allowed to cool prior to filtration through a pad of Celite and evaporation to dryness . The crude product was purified by column chromatography on silica gel to give 3.95 g (86% yield) of the stannane.

Synthesis of the B-Rincr

8

Synthesis of Diprotected 8

3-Hydroxybenzoic acid (24.86 g, 180 mmol) was dissolved in 600 mL of dry CH₂C1₂ under argon and cooled to 0°C. Triethylamine (56 mL, 400 mmol) was added. A solution of t-butyldimethylsilyl chloride (56.0 g, 370 mmol in 200 mL of dry CH₂C1₂) was added dropwise with stirring using a pressure equalized addition funnel and the reaction stirred for 12 hours. The mixture was the evaporated to dryness, redissolved in 400 mL of Et₂0, and washed with 3 x 100 mL portions of deionized water. The organic layer was dried with Na₂S0₄, concentrated, and used without further purification.

Conversion to the Acid Chloride 9

The silyl ester 8_. (11.3 g, 30 mmol) was dissolved in 100 mL of dry CH₂C1₂ containing 5 drops of DMF under argon. The solution was cooled to 0°C prior to dropwise addition of oxalyl chloride (10.5 mL, 120 mmol) . The reaction was allowed to warm to room temperature over 12 hours . The crude product was evaporated to approximately 25 mL and filtered through a plug of silica gel using 25 mL of CH₂C1₂ as a wash. The product was concentrated and used without further purification.

Synthesis of Resin Bound Benzophenone

Benzophenone 10

The stannane (6_.) (3.95 g, 10.1 mmol) was dissolved with stirring in 100 mL of dry THF under argon prior to addition of oven dried K₂CO₃(0.71 g, 5.0 mmol). Hunig's base (2.4 mL, 13.5 mmol) was then added with stirring. The acid chloride ( 9_) (2.70 g, 10 mmol) was dissolved in 50 mL of dry THF and slowly added to the stannane solution. Tris (dibenzylidene acetone) dipalladium (0) (0.50 g, 0.6 mmol) was added in one portion and the mixture was stirred at room temperature until formation of a palladium mirror was observed (approximately 20 minutes) . Diethyl ethyl ether (100 mL) was added and the mixture filtered through a pad of Celite. The solution was evaporated under reduced pressure and purified by column chromatography over silica gel to give 2.16 g (4.7 mmol, 47% yield) of benzophenone 10.

10 11

Reduction to Benzyl Alcohol 11

A solution of benzophenone 10 (2.16 g, 4.7 mmol) in THF (50 mL) was cooled under argon with stirring to 0°C. Excess lithium tri-t-butoxy aluminohydride (1.22 g, 4.8 mmol) was added in one portion and the mixture stirred for 3 hours. Deionized water (20 mL) was used to quench the reaction and the resulting mixture was extracted with 3 x 100 mL portions of diethyl ether. The combined organic fractions were dried with magnesium sulf te, concentrated under reduced pressure and purified by column chromatography on silica gel to give 1.94 g (89% yield) of the benzyl alcohol.

Synthesis of Acid Chloride Resin

Carboxylated polystyrene resin (30.0 g, 3.0 mmol/g) was heated to reflux in dry toluene (200 mL) under argon with excess oxalyl chloride (39.0 mL, 1.455 g/mL, 450 mmol) for 26 hours. The slurry was cooled, filtered under dry nitrogen, and washed with toluene (3 x 100 mL portions) . The resin was dried under high vacuum for 12 hours (2.57 mmol Cl by elemental analysis) .

Coupling to Resin

The benzyl alcohol 3L (1.94 g, 4.2 mmol), triethylamine (1.1 mL, 8.0 mmol), acid chloride resin (1.44 g, 2.57 mmol/g) and a catalytic amount of N,N- dimethylaminopyridine (15 mg) were combined with 50 mL of dry methylene chloride under argon in a resin ' reactor and shaken for 18 hours. The resin was filtered and sequentially washed with 2 x 20 mL portions of methylene chloride, THF, DMF, THF, and finally methylene chloride. The resin was then dried under high vacuum to 2.46 g of Y2_ (1.0 mmol/g) .

On Resin Manipulation

Deprotection of 12

The resin (^2_.) was deprotected in 10 hours using a stock TBAF solution (0.25 mL of a 1.0 M TBAF/THF solution) , drained, and washed with 1 mL portions of CH₂C1₂ (3), THF (3), DMF (3), THF (3), and CH₂C1₂ (3) to give the phenolic resin 13. This was then treated with 0.4 mL of a stock methyl alcohol solution (49 mL in 1.75 mL toluene) and 0.5 mL of freshly prepared Merck reagent (6.50 g suspended in 32 mL of CH₂C1₂) . The well was capped and rotated for 4 days prior to draining and washing with 1 mL portions of CH₂C1₂ (3), THF (3), DMF (3), THF (3), and CH₂Cl₂ (3) to give 14.

Hydride Reduction of Benzophenone (14)

Borane (5.0 mL, 1.0 M in THF) was added to a 50 mL round bottom flask containing dry THF (10 mL) under argon and cooled to 0°C. 2 -Methyl-2 -butene (5.0 mL, 2.0 M in THF) was added dropwise with stirring and the solution was stirred for 1 hour. The resulting disiamylborane solution was added in one portion to a 25 mL vial containing 500 mg of resin 14_.. After the evolution of gas had subsided, the mixture was capped and rotated overnight. Filtration and washing with CH₂C1₂ (2 X 20 mL) , 0.1 N HCl: THF (1:10) (1 X 20 mL) , THF (2 X 20 mL) , and finally CH₂C1₂ (2 X 20 mL) gave 500 mg of reduced resin 15.

Cleavage

A stock solution of NaOMe was prepared by dissolving sodium methoxide (0.47 g, 8.8 mmol) in methanol (12 mL) and diluting with THF (52 mL) . The resin was treated with the methoxide solution (0.95 mL) for hours without agitation. The well was drained and the filtrate evaporated to dryness. The residue was neutralized with excess HCl/Et₂0 (0.9 mL, 1.0 M) and once again evaporated to dryness to give .16_. (LC/ MS 73% pure, [M+H] = 365) .

EXAMPLE 2

General Procedures for Formula I Wherein M_η is -CH(NRR')- Derivazation of Resin 14

To a suspension of the resin 14 (50 mg, 0.05 mmol) in a mixture of THF and trimethylorthoformate (1/1, 5 ml) is added phenethyamine (10 eq.) . The mixture is stirred for 6 hours, prior to draining and washing with dry THF (2 x 10 ml) . The resin is again suspended in dry THF and treated for 12 hours with a 1 M solution in THF of sodium cyanoborohydride (10 eq. , 0.5 ml) . The resin is filtered and washed with THF (2 x 10 ml) , then with MeOH (2 x 10 ml) , followed again with THF (2 x 10 ml) , to give 17. Cleavage

Similar conditions as described above with regard to compound 1.6 are used to give 18.

EXAMPLE 3

Protein Kinase C Activity Determination Compounds of the invention are tested for ability to inhibit protein kinase C using rat brain as the enzyme source accordingly to widely used procedures such as described by A.C. McArdle and P.M. Conn, Methods in Enzymology (1989) 168, 287-301, and by U. Kikkawa et al . , Biochem. Biophys. Res. Commun. (1986), 135 , 636-634. Alternatively, protein kinase C activity is determined using purified human protein kinase C isozymes by methods such as described in P. Basta et al. , Biochim. Biophys. Acta. (1992) 1132, 154-160.

EXAMPLE 4

Membrane Receptor Affinity Determinations A. Bradykinin Receptor

The bradykinin receptor affinity of compounds prepared according to this invention is determined for ability to displace [³H] bradykinin binding from guinea pig ileal membrane as described in S.G. Farmer et al . , J.Pharmacol. Exp. Ther. (1989) 248, 677.

B. Other Receptors

Generally applicable methods for testing receptor affinity of the compounds of the invention are described by H.I. Yamamura et al . , Methods in Neurotransmitter Receptor Analysis, raven Press, 1990.

EXAMPLE 5

Membrane of Interaction with Target Enzymes

A. Angiotensin Converting Enzyme

Methods useful for determining the ability of compounds of the invention to inhibit angiotensin converting enzyme are disclosed by J.W. Ryan, Methods in Enzymology (1988) 164, 194-211.

B . Phospholipase A., A procedure useful to test efficacy of the invented compounds in inhibiting phospholipase A₂ is described by J. Reynolds et al . , Methods in Enzymology (1991) 197, 3-23.

EXAMPLE 6

Determination of Ion Channel Binding Xenopus oocytes are will known as tools for studying ion channels and receptors. A. L. Buller and M. M. White have described methods useful to measure interaction between compounds of the invention and various ion channels or receptors . Methods in Enzymology (1992) 207. 368-375. EXAMPLE 7

Transcription Factor Effects J. M. Gottesfeld is an example of a reference describing a procedure suitable for analyzing the ability of the invented compounds to influence transcription factor function. Methods in Enzymology (1977) 170, 346,359.

The disclosures of all references cited in this specification herein are incorporated in their entireties by reference.

The foregoing examples are illustrative of the present invention and are not to be construed as limiting thereof.

Claims

THAT WHICH IS CLAIMED IS:

A library of compounds having the formula

wherein: M₁ and M₂ independently are CH(NRR') or

CH(OR) , wherein R and R' independently are H, C₀- C₆alkylCOR₁₅,

C₁.₆alkylNR_2SR₂g, C₀.₆alkylNR₈₀C(NR₈₁)NR₈₂R₈₃, C^alkylindole, C₀.₆alkyl-D, C_x. ₆alkylCN, C₀_₆alkylSOR₁₅, C₀.₆alkylC (0) R₁₆, S00R₁₆, or C_x_ _galkylOC^_galkylR^;

X_1# Y.. and Z are any accessible combination of CH, CHCH, O, S, N, and NH provided that at least one is CH or CHCH and not more than one is CHCH;

X₂, Y₂ and Z₂ are any accessible combination of CH, CHCH, O, S, N, and NH provided that at least one is CH or CHCH and not more than one is CHCH;

V is H, C^alkyl, halo, (C₀.₄alkyl) OH, (C₀_ alkylSH, (C₀.₄alkyl) NR₂₂R₂₃, (C₀.₄alkyl) C0₂R₇₆, or C₀_ ₄alkylOR₇; W is H or

X₃, Y₃, and Z₃ are any accessible combination of CH, CHCH, 0, S, N, and NH provided that at least one is CH or CHCH and not more than one is CHCH; R₇, Rig, R ₀' ^₂2' R-23' ^₈₀' ^₈₁' ^₈2' ³ⁿ R_g3 independently are H or C_halky!; R₁₆, R₁₇, R₁₈, R₂₄, R₂₅, R₂₆, and R₇₆ independently are H, C^alkyl, phenyl, or substituted phenyl; R₁₅ is 0R₁₈ or NR₁₉R₂₀; and

^R22' ^-23' ^R75' K.76' ^77' ^an<3 ^78' independently are H or C_halky1.

2. The library of Claim 1, wherein the compounds have a molecular weight of between about 300 and about 600 daltons.

3. The library of Claim 1, wherein said compounds have the formula:

wherein :

X_x, Y_x and Z₁ are any accessible combination of CH, CHCH, 0, S, N, and NH provided that at least one is CH or CHCH and not more than one is CHCH;

X₂, Y₂ and Z₂ are any accessible combination of CH, CHCH, 0, S, N, and NH provided that at least one is CH or CHCH and not more than one is CHCH; W is H or

X₃, Y₃ and Z₃ are any accessible combination of CH, CHCH, 0, S, N, and NH provided that at least one is CH or CHCH and not more than one is CHCH; M_x and M₂ independently are CH(NRR') or CH(OR) , wherein R and R' independently are H, C₀- C₆alkylCOR_ls,

C₁.₆alkylNR₂₅R₂₆, C₀.₆alkylNR₈₀C(NR₈₁)NR₈₂R₈₃, C^alkylindole, C₀._salkyl-D, C_x. ₆alkylCN, C₀.₆alkylSOR₁₅, C₀.₆alkylC (O) R₁₆, SOOR₁₆, or C_λ_

V is H, C^alkyl, halo, (C₀.₄alkyl) OH, C(C₀. ₄alkyl)SH, (C₀.₄alkyl) NR₂₂R₂₃, (C₀.₄alkyl) C0₂R₇₆ or C₀_ ₄alkylOR₇; A₁₇ A₂, A₃, A₄, A₅, and A₆ independently are absent or present as 0, S, NR₆₀; or C₀.₆alkylC (0) NR₂₁, provided that at least two are present;

R_x, R₂, R₃, R₄, R₅, and R₆ independently are H, C₀__salkylCOR₁₅, C^_galkylR^R^, C^alkylOR^ except methoxymethyl , C₁.₆alkylNR₂₅R₂₆, C₀.₆alkylNR₈₀C (NR₈₁) NR₈₂R₈₃, C^alkylindole, C₀.₆alkyl-D, C^alkylCN, C₀.₆alkylSOR₁₅, or C^alkylOC^alkylR^ ;

D is one or multiple fused saturated or unsaturated five or six membered cyclic hydrocarbon or heterocyclic ring system containing one or two 0, N, or S atoms that is substituted or unsubstituted by any accessible combination of 1 to 4 substituents selected from C^alkyl, NR₇R₈, OR₉,SR₁₀, COR^, halogen, CF₃, or O;

R-7 ' ^8 ' "9' 10' "-19' R-20' "21' "22' "23' "^■SO' "80 ' R₈₁, R₈₂, and R₈₃ independently are H or

independently are H,

phenyl, or substituted phenyl ;

R_xl is 0R₁₂ or NR₁₃R₁₄; and R₁₅ is OR₁₈ or NR₁₉R₂₀.

4. The library of Claim 1, wherein in the compounds of the library, the X, Y, and Z containing ring is pyridine, pyrimidine, triazine, pyrrole, imidazole, oxazole, isoxazole, thiophene, thiazole, or furan .

5. The library of Claim 3 , wherein said compounds have the formula :

wherein :

M-_L is CH(O(C₀.₄alkyl)R) , wherein R is H, C_x. ₆alkyl, D, COOH, COOC^alkyl, OH, OC^alkyl, NH₂, N(C₁_ ₆alkyl)₂, or CN;

V_x is H, CH₃, OH, or CH₂OH; A₇, A₈, A₉, and A₁₀ independently are absent or present as O provided that two are 0; and

R₃₀, R₃₁, R₃₂, and R₃₃ independently are H, C_x. ₆alkyl, D, COOH, COOC^alkyl, OH, OC^alkyl, NH₂, N(C_X. ₆alkyl)₂, or CN.

6. The library of Claim 3 , wherein said compounds have the formula:

wherein:

M-_L and M₂ independently are CH(NRR') or CH(OR) , wherein R and R' independently are H, C₀-

C₆alkylCOR₁₅ , C^C_galkylR^R,, , C^_galkylOR^ , , C₁.₆alkylNR₂₅R₂₆ ,

C₀.₆alkylNR₈₀ C (NR₈₁) NR₈₂R₈₃ , C₀., _Salkyl -D , C^a ilkylCN, C₀.

₆alkylSOR₁₅ , C₀.₆alkylC (O) R₁₆ , SOOR₁₆ , or C . .₆alkylOC_x.

₆alkylR₁₅ ; B and B' are H, O (CH₂) _nNR₄₀C (NR₄₁) NR₄₂R₆₁, or O (CH₂)_n,NR₄₃R₄₄ wherein R₄₀, R₄₁, R₄₂, R₄₃, R₄₄, and R_S1, independently are H or C^alkyl, n and n' are 2 or 3; provided one of B and B' is H; E is

wherein X is CH, N, NH, 0, S; n is 1-3; n' is 1 when X is 0,S, or NH; and n' is 2 when X is CH or N; F, F', and F" are H, 0 (CH₂) _nNR₄₅C (NR₄₆) NR₄₇R₆₂, or 0(CH₂)_n,NR₄₈R₄₉ wherein R₄₅, R₄₆, R₄₇, R₄₈, R₄₉, and R₆₂ independently are H or C-^alkyl, and n and n' are 2 or 3; provided two of F, F',and F" are H;

G and G' are H, O(CH₂)_nOR₅₀, or

wherein X' is CH, N, NH, 0, or S';

R₅₀ is H or C_j^^alkyl; and

R₅₁ is H, C^alkyl, halogen, OH, or OC^alkyl; n is 1-3; and n' is 1 or 2; provided one of G and G' is H.

7. The library of Claim 3 , wherein said compounds have the formula :

wherein:

M_x and M₂ independently are CH(NRR') or CH(OR) , wherein R and R' independently are H, C₀-

C₆alkylCOR₁₅,

C₀.₆alkylNR₈₀C(NR₈₁)NR₈₂R₈₃, C^alkylindole, C₀_₆alkyl-D, C .

₆alkylCN, C₀_₆alkylSOR₁₅, C₀.₆alkylC (0) R₁₆, SOOR₁₆, or C _

J, J' , and M independently are

O(CH₂)_nNR₅₀C(NR₅₁)NR₅₂R_6S or 0 (CH₂) _n,NR₅₃R₅₄ wherein R₅₀, R₅₁,

R₅₂, R₅₃, R₅₄, R₆₅ independently are H or C^alkyl, and n and n¹ independently are 2-3;

Q and Q' are H or 0 (C^alkyl) T wherein T is C^alkyl, C0₂R₅₅, OR_S6, or

wherein:

X₇ is CH, N, NH, S, or 0; n' is 1 or 2;

U is H, C^alkyl, halogen, CF₃, or OR₅₇; and ^R _s5, ^R ₅₆' ^an( R₅₇ independently are H or C₁_ _salkyl; provided that at least Q or Q ' is H.

A library of compounds having the formula :

wherein:

X₁, Y_x and Z_x are any accessible combination of CH, CHCH, 0, S, N, and NH provided that at least one is CH or CHCH and not more than one is CHCH;

X₂, Y₂ and Z₂ are any accessible combination of CH, CHCH, 0, S, N, and NH provided that at least one is CH or CHCH and not more than one is CHCH;

W is H or

X₃, Y₃ and Z₃ are any accessible combination of CH, CHCH, 0, S, N, and NH provided that at least one is CH or CHCH and not more than one is CHCH;

M₁ and M₂ independently are CH(NRR') or CH(OR) , wherein R and R' independently are H, C₀-

C₆alkylCOR₁₅, C^ alkylR^R^, C^alkylOR^, C₁._salkylNR₂₅R₂₆, C₀._salkylNR₈₀C(NR₈₁)NR₈₂R₈₃, C^alkylindole, C₀._salkyl-D, C_x. ₆alkylCN, C₀_₆alkylSOR₁₅, C₀.₆alkylC (0) R₁₆, S00R₁₆, or C _ _galkylOC.^alkylR^; V is H, C^alkyl, halo, (C₀.₄alkyl) OH, C(C₀.

₄alkyl)SH, (C₀.₄alkyl) NR₂₂R₂₃, (C₀.₄alkyl) C0₂R₇₆ or C₀_ ₄alkylOR₇;

A_x, A₂, A₃, A₄, A₅, and A₆ independently are absent or present as 0, S, NR₆₀; or C₀.₆alkylC (0) NR₂₁, provided that at least two are present;

R_x, R₂, R₃, R₄, R₅, and R₆ independently are H, C₀.₆alkylCOR₁₅,

except methoxymethyl , C₁.₆alkylNR₂₅R₂₆ , C₀.₆alkylNR₈₀C (NR₈₁ ) NR₈₂R₈₃ ,

C₀.₆alkylSOR₁₅, or C^_galkylOC^_galkylR^; D is one or multiple fused saturated or unsaturated five or six membered cyclic hydrocarbon or heterocyclic ring system containing one or two 0, N, or S atoms that is substituted or unsubstituted by any accessible combination of 1 to 4 substituents selected from C_x.₆alkyl, NR₇R₈, OR₉,SR₁₀, COR_11# halogen, CF₃, or 0;

"7' "8' "9' "10' "19' "20' "21' "22' "23' "60' "80'

^Rβi' ^R ₈₂' ^and ^R ₈₃ independently are H or

^R12' ^R13' ^R14' ^R16' ^R17' ^R18 ' ^R24 ' ^R25 ' ^R26 ' ^a^ld R_7g independently are H,

phenyl, or substituted phenyl ;

R_X1 is OR₁₂ or NR₁₃R₁₄; and R₁₅ is 0R₁₈ or NR₁₉R₂₀.

9. The library of compounds of Claim 8, wherein X₁₇ Y_ιr X₂, X₃, and Y₃ are CH and Z_x, Z₂, and Z₃ are CHCH.

10. The library of compounds of Claim 9, wherein A_1# A₂, A₃, A₄, A₅, and A₆ are oxygen.

11. The library of compounds of Claim 8, wherein X_lf Y_lf X₂, Y₂ are CH, Z and Z₂ are CHCH, and W is H.

12. The library of compounds of Claim 11, wherein A_x, A₂, A₃, and A₄ are oxygen.

13. The library of compounds of Claim 8, having the following formula:

wherein:

M_x is CH(O(C₀.₄alkyl)R) , wherein R is H, C_x_ ₆alkyl, D, COOH, COOC₁__βalkyl, OH, OC^alkyl, NH₂, N(C,_ ₆alkyl) ₂, or CN; Vj_. is H, CH₃, OH, or CH₂OH;

A₇, A₈, A₉, and A₁₀ independently are absent or present as 0 provided that three are O; and

R₃₀, R₃₁, R₃₂, and R₃₃ independently are H, C₁_ ₆alkyl, D, COOH, COOC^alkyl, OH, OC^alkyl, NH₂, N(C_X. ₆alkyl) ₂, or CN.

14. The library of compounds of Claim 8, having the following formula:

wherein: M_x and M₂ independently are CH(NRR') or

CH(OR) , wherein R and R' independently are H, C₀- C₆alkylCOR₁₅, C.-C₈alkylR^R,,, C^alkylOR^, C₁.₆alkylNR₂₅R₂₆, C₀._ealkylNR₈₀C(NR₈₁)NR₈₂R₈₃, C₀.₆alkyl-D, ₀. ₆alkylSOR₁₅, C₀_₆alkylC (O) R₁₆, SOOR₁₆, o

₆alkylR₁₅ ;

B and B' are H, 0 (CH₂) _nNR₄₀C (NR₄₁) NR₄₂R₆₁, or O (CH₂)_n,NR₄₃R₄₄ wherein R₄₀, R₄₁, R₄₂, R₄₃, R₄₄, and R₆₁, independently are H or C^alkyl, n and n' are 2 or 3 ; provided one of B and B' is H; E is

wherein X is CH, N, NH, 0, S; n is 1-3; n' is 1 when X is 0,S, or NH; and n' is 2 when X is CH or N;

F, F', and F" are H, O (CH₂) _nNR₄₅C (NR_4S) NR₄₇R₆₂, or 0(CH₂)_n,NR₄₈R₄₉ wherein R₄₅, R₄₆, R₄₇, R₄₈, R₄₉, and R₆₂ independently are H or C₁.₃alkyl, and n and n' are 2 or 3; provided two of F, F ' , and F" are H;

G and G' are H, O(CH₂)_nOR₅₀, or

wherein X' is CH, N, NH, 0, or S¹;

R₅₀ is H or C^alkyl; and

R₅₁ is H, C^alkyl, halogen, OH, or OC^alkyl; n is 1-3; and n' is 1 or 2; provided one of G and G' is H.

15. The library of compounds of Claim 14 , wherein:

B or B' is OCH₂CH₂NHC(NH)NH₂; E is

or

F and F" are OCH₂CH₂NHC (NH) NH₂; and G and G' are H.

16. The library of compounds of Claim 8, having the following formula:

wherein:

M_x and M₂ independently are CH(NRR') or CH(OR) , wherein R and R' independently are H, C₀-

C₆alkylCOR₁₅, C₁-C₆alkylR₁₆R₁₇, C^alkylOR^, C^alkylNR^R^, C₀_₆alkylNR₈₀C(NR₈₁)NR₈₂R₈₃, C^alkylindole, C₀_₆alkyl-D, C_x_ ₆alkylCN, C₀.₆alkylSOR₁₅, C₀.₆alkylC (0) R₁₆, S00R₁₆, or C_x.

J, J' , and M independently are

O(CH₂)_nNR₅₀C(NR₅₁)NR₅₂R₆₅ or 0 (CH₂) _n,NR₅₃R₅₄ wherein R₅₀, R₅₁, R₅₂, R₅₃, R₅₄, R₆₅ independently are H or C₁.₃alkyl, and n and n' independently are 2-3;

Q and Q' are H or 0 (C^alkyl) T wherein T is C_halky!, C0₂R₅₅, OR₅₆, or

wherein:

X₇ is CH, N, NH, S, or O; n' ' ' is 1 or 2;

U is H,

halogen, CF₃, or OR₅₇; and R_55/ R₅₆, and R₅₇ independently are H or C₁_ ₆alkyl; provided that at least Q or Q' is H.

17. A library of compounds having the formula :

wherein: X_ιr Y_x and Z are any accessible combination of CH, CHCH, 0, S, N, and NH provided that at least one is CH or CHCH and not more than one is CHCH;

X₂, Y₂ and Z₂ are any accessible combination of CH, CHCH, 0, S, N, and NH provided that at least one is CH or CHCH and not more than one is CHCH; W is H or

X₃, Y₃ and Z₃ are any accessible combination of CH, CHCH, 0, S, N, and NH provided that at least one is CH or CHCH and not more than one is CHCH;

M₁ and M₂ independently are CH(NRR') or CH(OR) , wherein R and R' independently are H, C₀- C₆alkylCOR₁₅,

C₀.₆alkylNR₈₀C(NR₈₁)NR₈₂R₈₃,

C₀.₆alkyl-D, C_x_ _GalkylCN, C₀.₆alkylSOR₁₅, C₀.₆alkylC (0) R₁₆, S00R₁₆, or C_x.

V is a bond to a solid support; A₁₇ A₂, A₃, A₄, A₅, and A₆ independently are absent or present as 0, S, NR₆₀; or C₀.₆alkylC (0) NR₂₁, provided that at least two are present ; R₁₇ R₂, R₃, R₄, R₅, and R₆ independently are H,

C₀.₆alkylCOR₁₅,

except methoxymethyl ,

C₀.₆alkylNR₈₀C (NR₈₁) NR₈₂R₈₃, Ci.galkylindole, C₀_₆alkyl-D, C^alkylCN, C₀.₆alkylSOR₁₅, or

D is one or multiple fused saturated or unsaturated five or six membered cyclic hydrocarbon or heterocyclic ring system containing one or two 0, N, or S atoms that is substituted or unsubstituted by any accessible combination of 1 to 4 substituents selected from C^alkyl, NR₇R₈, OR₉,SR₁₀, COR_lx, halogen, CF₃, or 0;

"7/ "8' "9' "10' "19' "20' "21' "22' "23' "60' "80'

R₈₁, R₈₂, and R₈₃ independently are H or C₁.₆alkyl ;

^R12' ^R13' ^R14' ^R16' ^R17' ^R18 ' ^R24 ' ^R25 ' ^R26 ' ^a^d R_7g independently are H,

phenyl, or substituted phenyl ;

R_π is 0R₁₂ or NR₁₃R₁₄; and R₁₅ is 0R₁₈ or NR₁₉R₂₀.

18. The library of compounds of Claim 17, wherein X_lf Y_lt X₂, Y₂, X₃, and Y₃ are CH and Z_x, Z₂, and Z₃ are CHCH.

19. The library of compounds of Claim 18, wherein A_ι; A₂, A₃, A₄, A_s, and A₆ are oxygen.

20. The library of compounds of Claim 17, wherein X_lf Y_ιr X₂, Y₂ are CH, Z_x and Z₂ are CHCH, and W is H or Ci.galkyl.

21. The library of compounds of Claim 20, wherein A_lt A₂, A₃, and A₄ are oxygen.

22. A method for preparing a combinatorial library of bridged biaromatic and triaromatic ring compounds of Formula I :

wherein:

X_λ, Y and Z₁ are any accessible combination of CH, CHCH, 0, S, N, and NH provided that at least one is CH or CHCH and not more than one is CHCH;

X₂, Y₂ and Z₂ are any accessible combination of CH, CHCH, 0, S, N, and NH provided that at least one is CH or CHCH and not more than one is CHCH;

W is H or

X₃, Y₃ and Z₃ are any accessible combination of CH, CHCH, 0, S, N, and NH provided that at least one is CH or CHCH and not more than one is CHCH;

M₁ and M₂ independently are CH(NRR') or CH(OR) , wherein R and R' independently are H, C₀-

C₆alkylCOR₁₅,

C₀.₆alkylNR₈₀C(NR₈₁)NR₈₂R₈₃,

C₀.₆alkyl-D, C_x_ ₆alkylCN, C₀.₆alkylSOR₁₅, C₀._salkylC (0) R₁₆, S00R₁₆, or C_x.

V is a moiety which can be bonded to a cleavable linker of a solid support and is selected from the group consisting of H,

halo, (C₀_ ₄alkyl)OH, C (C₀_₄alkyl) SH, (C₀.₄alkyl) NR₂₂R₂₃, (C₀_ ₄alkyl) C0₂R₇₆ and C₀.₄alkylOR₇;

A_1# A₂, A₃, A₄, A₅, and A₆ independently are absent or present as 0, S, NR₆₀; or C₀.₆alkylC (0) NR₂₁, provided that at least two are present;

R_x, R₂, R₃, R₄, R₅, and R₆ independently are H, C₀.₆alkylCOR₁₅,

except methoxymethyl , C₁.₆alkylNR₂₅R_2g, C₀.₆alkylNR₈₀C (NR₈₁) NR₈₂R₈₃, C^alkylindole, C₀.₆alkyl-D, C^alkylCN, C₀.₆alkylSOR₁₅, or C^_galkylOC^_galkylR^ ;

D is one or multiple fused saturated or unsaturated five or six membered cyclic hydrocarbon or heterocyclic ring system containing one or two 0, N, or S atoms that is substituted or unsubstituted by any accessible combination of 1 to 4 substituents selected from C^_galkyl, NR₇R₈, OR₉,SR₁₀, C0R₁₁₇ halogen, CF₃, or 0;

"7' "8' "9' "10' "19' "20' "21' "22' "23' "60' "80'

^Rβi' ^Rβ₂'

^R12' 13' ^R14' 16' ^R17' ^R18 ' ^R24 ' ^R25 ' ^R26 ' ^and R₇g independently are H,

phenyl, or substituted phenyl ;

R_X1 is 0R₁₂ or NR₁₃R₁₄; and

R₁₅ is 0R₁₈ or NR₁₉R₂₀, wherein each library compound is made in a separate reaction zone, said process comprising the steps of: coupling a first scaffold of the formula

to a second scaffold of the formula

wherein V, X_lf Y_x, Z_x , X₂, Y₂, Z₂, A_XR₁₇ A₂R₂, A₃R₃, A₄R₄ and W have the meanings ascribed above and X and Y are moieties that react with one another to bond said first and second scaffolds; coupling said bonded scaffolds to a solid phase support with a cleavable linker; reacting said coupled scaffolds with a reagent suitable to form M_x; and cleaving said coupled scaffolds from said linker to form compounds of Formula I .

23. A method of screening for biologically active compounds, comprising the steps of: contacting a solid support containing a plurality of compounds physically separated from each other with an assay kit; and determining if any of the plurality of compounds has biological activity, wherein the plurality of compounds are of the formula :

wherein :

M-_L and M₂ independently are CH (NRR ' ) or CH (OR) , wherein R and R ' independently are H, C₀- C₆alkylCOR₁₅ ,

, C₁.₆alkylNR₂₅R₂₆ , C_0.6alkylNR₈₀C (NR₈₁) NR₈₂R₈₃ , C^alkylindole , C₀.₆alkyl-D, C^ ₆alkylCN, C₀.₆alkylSOR₁₅ , C₀.₆alkylC (0) R₁₆ , S00R₁₆ , or C_x_ _galkylOC^_galkylR^ ;

X_{l f} Y-, and Z_x are any accessible combination of CH, CHCH, 0, S , N, and NH provided that at least one is CH or CHCH and not more than one is CHCH; X₂, Y₂ and Z₂ are any accessible combination of CH, CHCH, 0, S, N, and NH provided that at least one is CH or CHCH and not more than one is CHCH;

V is H, C^alkyl, halo, (C₀.₄alkyl) OH, (C₀_ alkylSH, (C₀_₄alkyl) NR₂₂R₂₃, (C₀.₄alkyl) C0₂R₇₆, or C₀_ ₄alkylOR₇;

W is H or

X₃, Y₃, and Z₃ are any accessible combination of CH, CHCH, O, S, N, and NH provided that at least one is CH or CHCH and not more than one is CHCH;

"7/ "19' "20' "22' "23' "80' "81' "82' anC. Kg₃ independently are H or

R₁₆, R₁₇, R₁₈, R₂₄, R₂₅, R₂₆, and R₇₆ independently are H,

phenyl, or substituted phenyl;

R₁₅ is OR₁₈ or NR₁₉R₂₀; and 22' ^R23' 75' 76' ^R77' ^an<3- 7β ' independently are H or C_halky1.

24. An apparatus suitable as a replacement element in an automated assay instrument as a source of individual members of a library of structurally related compounds, said apparatus comprising a two-dimensional array of defined reservoirs, each reservoir containing a unique compound of said library, wherein said structurally related compounds are of the formula:

wherein: M.. and M₂ independently are CH(NRR') or CH(OR) , wherein R and R' independently are H, C₀- C_salkylCOR₁₅,

C₁.₆alkylNR_2SR₂₆, C₀__galkylNR₈₀C(NR₈₁)NR₈₂R₈₃, C^alkylindole, C₀.₆alkyl-D, C_x_ ₆alkylCN, C₀.₆alkylSOR₁₅, C₀.₆alkylC (0) R₁₆, S00R₁₆, or C_λ_

X₁₇ Y₁ and Z-. are any accessible combination of CH, CHCH, 0, S, N, and NH provided that at least one is CH or CHCH and not more than one is CHCH; X₂, Y₂ and Z₂ are any accessible combination of CH, CHCH, 0, S, N, and NH provided that at least one is CH or CHCH and not more than one is CHCH;

V is H, C^alkyl, halo, (C₀.₄alkyl) OH, (C₀_ alkylSH, (C₀_₄alkyl) NR₂₂R₂₃, (C„.₄alkyl) C0₂R₇₆, or C₀_ ₄alkylOR₇;

W is H or

X₃, Y₃, and Z₃ are any accessible combination of CH, CHCH, 0, S, N, and NH provided that at least one is CH or CHCH and not more than one is CHCH;

"77 "19' "20' "22' "23' "80' "81' "82' ^aU^Q _g3 independently are H or

R₁₆, R₁₇, R₁₈, R₂₄, R₂₅, R₂₆, and R₇₆ independently are H, C-^_galkyl, phenyl, or substituted phenyl; R₁₅ is OR₁₈ or NR₁₉R₂₀; and

R₂₂, R₂₃, R₇₅, R₇₆, R₇₇, and R₇₈, independently are H or C_halky1.

25. The apparatus of Claim 24, wherein the library compound in each reservoir has the formula

wherein:

X₁₇ Y_x and Z_x are any accessible combination of CH, CHCH, 0, S, N, and NH provided that at least one is CH or CHCH and not more than one is CHCH;

X₂, Y₂ and Z₂ are any accessible combination of CH, CHCH, 0, S, N, and NH provided that at least one is CH or CHCH and not more than one is CHCH;

W is H or

X₃, Y₃ and Z₃ are any accessible combination of CH, CHCH, 0, S, N, and NH provided that at least one is CH or CHCH and not more than one is CHCH;

M_x and M₂ independently are CH(NRR') or CH(OR) , wherein R and R' independently are H, C₀-

C₆alkylCOR₁₅, C₁-C₆alkylR₁₆R₁₇, C^alkylOR^, C^alkylNR^R^, C₀_₆alkylNR₈₀C(NR₈₁)NR₈₂R₈₃, C^alkylindole, C₀.₆alkyl-D, C _ ₆alkylCN, C₀.₆alkylSOR₁₅, C₀.₆alkylC (0) R₁₆, S00R₁₆, or C^ βalkylOCi.galkylRis ; V is H, C-^alkyl, halo, (C₀.₄alkyl) OH, C(C₀.

₄alkyl)SH, (C₀.₄alkyl) NR₂₂R₂₃, (C₀.₄alkyl) C0₂R₇₆ or C₀_ ₄alkylOR₇;

A₁₇ A₂, A₃, A₄, A₅, and A₆ independently are absent or present as 0, S, NR₆₀; or C₀.₆alkylC (0) NR₂₁, provided that at least two are present;

R₁₇ R₂, R₃, R₄, R₅, and R₆ independently are H, C₀.₆alkylCOR₁₅,

C₁_₆alkylOR₂₄ except methoxymethyl , C₁.₆alkylNR₂₅R₂₆ , C₀.₆alkylNR₈₀C (NR₈₁ ) NR₈₂R₈₃ ,

C₀.₆alkylSOR₁₅, or Ci.galkylOCi.galkylRis,-

D is one or multiple fused saturated or unsaturated five or six membered cyclic hydrocarbon or heterocyclic ring system containing one or two 0, N, or S atoms that is substituted or unsubstituted by any accessible combination of 1 to 4 substituents selected from C^alkyl, NR₇R₈, OR₉,SR₁₀, COR₁₁₇ halogen, CF₃, or 0; ₇ , g , Kg , K₁₀ , ₁₉ , K₂₀ , ₂₁ , K₂₂ , "₂₃ ' "6₀ ' "₈₀ '

R₈₁ , R₈₂ , and R₈₃ independently are H or

;

"12 ' "13 ' "14 ' "16 ' "17 ' "18 ' "24 ' "25 ' "26 ' an^Q ₇g independently are H, C^alkyl, phenyl, or substituted phenyl ; R_λl is OR₁₂ or NR₁₃R₁₄; and

R₁₅ is OR₁₈ or NR₁₉R₂₀, and is prepared in accordance with the process of Claim 22 and wherein each reservoir provides one reaction zone .

26. The apparatus of Claim 24, wherein said apparatus comprises a multi-well microtiter plate.

27. The apparatus of Claim 26, wherein said apparatus comprises a 96 well microtiter plate.

28. An assay kit for identification of pharmaceutical lead compounds, said kit comprising biological assay materials and a well plate apparatus, wherein each well in said apparatus contains a unique library compound of Claim 1.

29. The assay kit of Claim 28, containing biological materials for performing assay tests selected from the group consisting of in vitro assays, cell based functional assays, and add, incubate and read assays.

30. A pharmaceutically useful compound having the formula:

wherein:

X₁₇ Y-. and Z_x are any accessible combination of CH, CHCH, 0, S, N, and NH provided that at least one is CH or CHCH and not more than one is CHCH;

X₂, Y₂ and Z₂ are any accessible combination of CH, CHCH, 0, S, N, and NH provided that at least one is CH or CHCH and not more than one is CHCH; W is H or

X₃, Y₃ and Z₃ are any accessible combination of CH, CHCH, 0, S, N, and NH provided that at least one is CH or CHCH and not more than one is CHCH; _x and M₂ independently are CH(NRR') or

CH(OR) , wherein R and R' independently are H, C₀- C₆alkylCOR₁₅,

C₁.₆alkylNR₂₅R_2S, C₀.₆alkylNR₈₀C(NR₈₁)NR₈₂R₈₃, C^alkylindole, C₀.₆alkyl-D, C_x. ₆alkylCN, C₀.₆alkylSOR₁₅, C₀._salkylC (0) R₁₆, S00R₁₆, or C_x_

V is H, C^alkyl, halo, (C₀.₄alkyl) OH, C(C₀. ₄alkyl)SH, (C₀_₄alkyl) NR₂₂R₂₃, (C₀_₄alkyl) C0₂R₇₆ or C₀_ ₄alkylOR₇;

A₁₇ A₂, A₃, A₄, A₅, and A₆ independently are absent or present as 0, S, NR₆₀; or C₀.₆alkylC (0) NR₂₁, provided that at least two are present; R_1# R₂, R₃, R₄, R₅, and R₆ independently are H, C₀_₆alkylCOR₁₅,

except methoxymethyl , C₁_₆alkylNR₂₅R₂₆, C₀.₆alkylNR₈₀C (NR₈₁) NR₈₂R₈₃,

C₀.₆alkylSOR₁₅, or C^_galkylOC^_galkylR^;

D is one or multiple fused saturated or unsaturated five or six membered cyclic hydrocarbon or heterocyclic ring system containing one or two 0, N, or S atoms that is substituted or unsubstituted by any accessible combination of 1 to 4 substituents selected from C^alkyl, NR₇R₈, OR₉,SR₁₀, C0R_lι; halogen, CF₃, or O;

"7' "8' "9' "10' "19' "20' "21' "22' "23' "60' "80'

R₈₁, R₈₂, and R₈₃ independently are H or C₁_₆alkyl;

^R12' ^R13' ^R14' 16' ^R17' ^R18 ' ^R24 ' ^R25 ' ^R26 ' and R_?g independently are H, C₁_₆alkyl, phenyl, or substituted phenyl ;

R is OR₁₂ or NR₁₃R₁₄; R₁₅ is OR₁₈ or NR₁₉R₂₀; or pharmaceutically useful salts thereof.

31. A compound having the following formula:

wherein:

X₁₇ Y-_L and Z_x are any accessible combination of CH, CHCH, 0, S, N, and NH provided that at least one is CH or CHCH and not more than one is CHCH;

X₂, Y₂ and Z₂ are any accessible combination of CH, CHCH, O, S, N, and NH provided that at least one is CH or CHCH and not more than one is CHCH; W is H or

X₃, Y₃ and Z₃ are any accessible combination of CH, CHCH, 0, S, N, and NH provided that at least one is CH or CHCH and not more than one is CHCH;

M_x and M₂ independently are CH(NRR') or CH(OR) , wherein R and R' independently are H, C₀- C₆alkylCOR₁₅,

C₁.₆alkylOR₂₄, C₁_₆alkylNR₂₅R₂g, C₀_₆alkylNR₈₀C(NR₈₁)NR₈₂R₈₃, C^alkylindole, C₀.₆alkyl-D, C₁_ _εalkylCN, C₀.₆alkylSOR₁₅, C₀.₆alkylC (0) R₁₆, S00R₁₆, or C_x_ _βalkylOCi.galkylRi_s ;

V is H, C^alkyl, halo, (C₀.₄alkyl) OH, C(C₀. ₄alkyl)SH, (C₀.₄alkyl) NR₂₂R₂₃, (C₀.₄alkyl) C0₂R₇₆, or C₀_ ₄alkylOR₇ or a bond to a solid support; A₁₇ A₂, A₃, A₄, A₅, and A₆ independently are absent or present as 0, S, NR₆₀; or C₀.₆alkylC (0) NR₂₁, provided that at least two are present ;

R'₁₇ R'₂, R'₃, R'₄, R'₅ and R'₆ independently are a protecting group or H, C₀.₆alkylCOR₁₅, C_x. ₆alkylR₁₆R₁₇, C₁._ealkyl0R₂₄ except methoxymethyl , C_x_

₆alkylNR_2SR₂₆ , C₀.₆alkylNR₈₀C (NR₈₁)NR₈₂R₈₃, C^alkylindole, C₀_ ₆alkyl-D,

₆alkylR₁₅, provided that at least one of R' ₁ to R'₆ is a protecting group; D is one or multiple fused saturated or unsaturated five or six membered cyclic hydrocarbon or heterocyclic ring system containing one or two 0, N, or S atoms that is substituted or unsubstituted by any accessible combination of 1 to 4 substituents selected from

halogen, CF₃, or 0;

"7' "8' "9' "10' "19' "20' "21' "22' "23' "60' "80'

R₈₁, R₈₂, and R₈₃ independently are H or C_halky!; ^R12' ^R13' ^R14' ^R16' ^R17' 18 ' ^R24 ' ^R25 ' 26 ' ^and R_?6 independently are H,

phenyl, or substituted phenyl ;

R_1X is OR₁₂ or NR₁₃R₁₄; and R₁₅ is OR₁₈ or NR₁₉R₂₀.