US20040039002A1

US20040039002A1 - 6,7-Dimethoxyquinazolines and therapeutic use thereof

Info

Publication number: US20040039002A1
Application number: US10/454,960
Authority: US
Inventors: Fatih Uckun; Xing-Ping Liu; Rama Narla
Original assignee: Parker Hughes Institute
Current assignee: Parker Hughes Institute
Priority date: 1999-03-19
Filing date: 2003-06-05
Publication date: 2004-02-26
Also published as: JP2002540103A; NO20014560L; US20020137757A1; KR20020011973A; MXPA01009453A; EP1163228A1; WO2000056720A1; NO20014560D0; US6258820B1; AU4171900A; US7038049B2; US6358962B2; US6638939B2; US20050075353A1; US20010016588A1; CA2367861A1; IL144954A0

Abstract

Quinazoline compounds and methods for the treatment of cancer and for the treatment of allergic reactions.

Description

FIELD OF THE INVENTION

This application relates to quinazoline compounds, compositions and therapeutic methods for the treatment of cancers and treatment of allergic disorders by administering quinazoline compounds.

BACKGROUND OF THE INVENTION

Quinazoline compounds have been suggested as useful compounds in the treatment of cell growth and differentiation characterized by activity of the human epidermal growth factor receptor type2 (HER2). See, for example, Myers et al., U.S. Pat. No. 5,721,237. Some quinazoline derivatives have been suggested as useful as anti cancer agents for the treatment of specific receptor tyrosine kinase-expressing cancers, especially those expressing epithelial growth factor (EGF) receptor tyrosine kinase. See, for example, Barker et al., U.S. Pat. No. 5,457,105. It is generally taught that quinazolines exert their anti-tumor effects via tyrosine kinase inhibition. However, while some quinazoline compounds inhibit the growth of brain tumor cells, others with equally potent tyrosine kinase inhibitory activity fail to do so (Naria et.al., 1998, Clin Cancer Res. 4:1405-1414; Naria et al., 1998, Clin Cancer Res. 4:2463-2471).

Several tumors expressing EGF receptors are not killed by quinazoline compounds, whereas some tumors not expressing EGF recptors are. Thus, the cytotoxic activity of quinazoline compounds cannot be attributed to the compound's tyrosine kinase inhibitory activity, and particularly not to the compound's ability to inhibit EGF receptor tyrosine kinase. A chemical structure-activity relationship determining the anti-cancer activity of quinazoline derivatives has not been established.

Novel quinazoline compounds may provide potent new therapeutic molecules for the treatment of disorders such as cancers. Methods of using both known and novel quinazoline compounds that employ an understanding of structure-function relationships are needed.

SUMMARY OF THE INVENTION

A series of quinazoline compounds were synthesized and analyzed for therapeutic activities, including anti cancer activities, particularly against EGR receptor-negative leukemias. Specific quinazoline compounds of the invention were found to possess potent and specific tyrosine kinase inhibitory activities affecting cell proliferation and survival. Quinazoline compounds of the invention are demonstrated as useful for the treatment of specific tumors, including breast tumors, brain tumors, and leukemias, particularly EGF receptor-negative leukemias, and to be particularly useful in the treatment of multi rug resistant leukemias.

The invention provides novel quinazoline compounds of formula I, as disclosed below, as well as therapeutic methods utilizing these compounds.

BRIEF DESCRIPTION OF THE DRAWINGS

FIGS. [0007] 1A-1C are graphs showing cytotoxic activity of fluoro-substituted dimethoxy quinazoline compounds (F-dmQ) against leukemic NALM-6 cells.
FIGS. [0008] 2A-2C are graphs showing cytotoxic activity of F-dmQ on breast cancer BT-20 cells.
FIGS. [0009] 3A-3F are photographs showing induction of apoptosis in cancer cells by F-dmQ.
FIG. 4 is a bar graph showing adhesive properties of various cancer cells to extracellular matrix proteins. [0010]
FIGS. [0011] 5A-5F are bar graphs showing the effect of F-dmQ on cancer cell-adhesion to extracellular matrix (ECM) proteins.
FIGS. [0012] 6A-6E are photographs showing the effects of HI-P353 and HI-P364 on glioblastoma cell migration from spheroids.
FIGS. 7A and 7B are bar graphs showing the anti-invasive activity of fluoro-substituted quinazoline compounds (F-dmQ) against glioblastoma U373 and breast cancer MDA-MB-231 cells. [0013]
FIGS. [0014] 8A-8D are photographs showing depolymerizaton of actin stress fibers and microtubules by HI-P353.
FIGS. [0015] 9A-9I are photographs showing inhibition of actin stress fiber formation in glioblastoma cells by HI-P154.
FIGS. [0016] 10A-10C are graphs showing the inhibition of cancer cell growth in vivo by the quinazolines of the invention.

DETAILED DESCRIPTION OF THE INVENTION

Definitions: [0017]
The terms “quinazoline”, “quinazoline compound”, and “quinazoline derivative” are used interchangeably in this application to mean compounds of formula I. All scientific and technical terms used in this application have meanings commonly used in the art unless otherwise specified. As used in this application, the following words or phrases have the meanings specified. [0018]
Halo is fluoro, chloro, bromo, or iodo. Alkyl, alkanoyl, etc., denote both straight and branched groups; but reference to an individual radical such as “propyl” embraces only the straight chain radical, a branched chain isomer such as “isopropyl” being specifically referred to. (C[0019] ₁-C₄)alkyl includes methyl, ethyl, propyl, isopropyl, butyl, iso-butyl, and sec-butyl; (C₁-C₄)alkoxy includes methoxy, ethoxy, propoxy, isopropoxy, butoxy, iso-butoxy, and sec-butoxy; and (C₁-C₄)alkanoyl includes acetyl, propanoyl and butanoyl.
As used herein, “pharmaceutically acceptable carrier” means any material which, when combined with the compound of the invention, allows the compound to retain biological activity, such as the ability to potentiate antibacterial activity of mast cells and macrophages. Examples include, but are not limited to, any of the standard pharmaceutical carriers such as a phosphate buffered saline solution, water, emulsions such as oil/water emulsions, and various types of wetting agents. Compositions comprising such carriers are formulated by well known conventional methods (see, for example, Remington's Pharmaceutical Sciences, Chapter 43, 14th Ed., Mack Publishing Co., Easton, Pa.). [0020]
The term “conjugate” means a compound formed as a composite between two or more molecules. More specifically, in the present invention, the quinazoline derivative is bonded, for example, covalently bonded, to cell-specific targeting moieties forming a conjugate compound for efficient and specific delivery of the agent to a cell of interest. [0021]
The phrase “targeting moiety” means a molecule which serves to deliver the compound of the invention to a specific site for the desired activity. Targeting moieties include, for example, molecules that specifically bind molecules on a specific cell surface. Such targeting moieties useful in the invention include anti-cell surface antigen antibodies. Cytokines, including interleukins and factors such as granulocyte/macrophage stimulating factor (GMCSF) are also specific targeting moieties, known to bind to specific cells expressing high levels of their receptors. [0022]
The term “prodrug moiety” is a substitution group which facilitates use of a compound of the invention, for example by facilitating entry of the drug into cells or administration of the compound. The prodrug moiety may be cleaved from the compound, for example by cleavage enzymes in vivo. Examples of prodrug moieties include phosphate groups, peptide linkers, and sugars, which moieties can be hydrolyzed in vivo. [0023]
“inhibit” means to reduce by a measurable amount, or to prevent entirely. [0024]
“to treat” means to inhibit or block at least one symptom that characterizes a pathologic condition, in a mammal threatened by, or afflicted with, the condition. [0025]
Compounds of the Invention [0026]
Compounds of the invention include quinazolines having the formula: [0027]
where: [0028]
R[0029] ⁸is iodo; (C₁-C₄)hydroxyalkyl, methylenedioxy, ethylenedioxy, benzyloxy, OCF₃. SCF₃, SO₃H, SO₂F, SO₂NR²R³in which R²is hydrogen or (C₁-₄)alkyl and R³is hydrogen, (C₁-C₄)alkyl, or phenyl, NR²R⁴in which R²is as defined above and R⁴is phenyl, or R^aa group of the formula
in which R[0030] ⁵and R⁶are each, independently, hydrogen, (C₁-C₄)alkyl or (C₁-C₄)perfluoroalkyl, and R⁷is hydrogen, halo, hydroxy, (C₁-C₄)alkyl, (C₁-C₄)alkoxy, (C₁-C₄)hydroxyalkyl, or N(R²)₂in which R²is as defined above;
n is an integer of 1-4; [0031]
R[0032] ^bis each, independently, hydrogen; halo; hydroxy; mercapto; (C₁-C₄)alkyl, (C₁-C₄)alkoxy, (C₁-C₄)thioalkyl, (C₁-C₄)hydroxyalkyl, nitro, cyano, methylenedioxy, ethylenedioxy, COCH₃, CF₃, OCF₃; SCF₃; COOH; SO₃H; SO₂F; phenyl or phenyl substituted by a group selected from halo, hydroxy, mercapto, (C₁-C₄)alkyl, (C₁-C₄)alkoxy, (C₁-C₄)thioalkyl, (C₁-C₄)hydroxyalkyl, amino, nitro, cyano, CF₃, COOH, SO₃H, SO₂NR²R³in which R²and R³are as defined below, and SO₂F. R^acan also be benzyloxy or benzyloxy substituted on the phenyl portion by a group defined above, NR²R³in which R²is H or (C₁-C₄)alkyl and R³is H, (C₁-C₄)alkyl, phenyl or phenyl substituted by a group as defined above;
R[0033] ¹is (C₁-C₄)alkyl, preferably methyl, or a pharmaceutically acceptable salt thereof, such as an acid addition salt.
Preferably, R[0034] ^ais a member selected from the group consisting of I, NHC6H5, —OCH₂CH₂O—, —OCH₂O—, OCF₃, SCF₃, CH₂OH, C₂H40H, SO₃H, SO₂NH₂, and SO₂F; and ore preferably R^ais I OCF₃or SO₂F. Most preferably, R^ais I or R^ais OCF₃.
In an alternative preferred compound, n is I and R[0035] ^ais a group of the formula:
Preferably, R[0036] ⁵and R⁶are each, independently, H, CH₃or CF₃, and most preferably, R⁵and R⁶are CF₃and R⁷is NH₂.
In another preferred compound, R[0037] ^bis at least one member selected from the group consisting of F, Cl, Br, I, OH, NH₂, NO₂, CN, COOH, CH₃, and CF₃, and more preferably R^bis at least one member selected from the group consisting of F, Cl, Br, OH, and CF₃.
Additional preferred quinazoline compounds useful in the treatment of tumors are described more fully below and particularly in the Examples. These include: [0038]
4-(3′,5′-dibromo-4′-methylphenyl)amino-6,7-dimethoxyquinazoline; [0039]
4-(2′,4′,6′-tribromophenyl)amino-6,7-dimethoxyquinazoline; [0040]
4-(2′,3′,5′,6′-tetrafluoro-5′-bromophenyl)amino-6,7-dimethoxyquinazoline; [0041]
4-(4′-fluorophenyl)amino-6,7-dimethoxyquinazoline; [0042]
4-(4′trifluoromethylphenyl)amino-6,7-dimethoxyquinazoline; and [0043]
4-(3′,5′-bis-trifluoromethylphenyl)amino-6,7-dimethoxyquinazoline. [0044]
Methods of Treatment [0045]
The compounds of the invention are useful for the treatment of animals, including humans. In particular, the compounds of the invention have been found to be potent inhibitors of tumor cell proliferation and survival, and effective to induce apoptosis of malignant cells. [0046]
Compounds of the invention have surprisingly been found to be effective for inducing apoptosis and/or cytotoxicity of leukemia cells. In particular, 4-(4′-hydroxyphenyl)amino-6,7-dimethoxyquinazoline compounds of the invention have been found to effectively induce apoptosis in multi-drug resistant leukemia. A preferred compound for the treatment of multi-drug resistant leukemia is 4-(3′-bromo-4′-hydroxyphenyl)amino-6,7-dimethoxyquinazoline. [0047]
Compounds of the invention that are particularly useful for treating leukemia include: [0048]
4-(3′,5′-bromo-4-methylphenyl)amino-6,7-dimethoxyquinazoline, [0049]
4-(2′,4′,6′-tribromophenyl)amino-6,7-dimethoxyquinazoline, [0050]
4-(2′,3′,5′,6′-tetrafluoro-4′-bromophenyl)amino-6,7-dimethoxyquinazoline, [0051]
4-(4′-fluorophenyl)amino-6,7-dimethoxyquinazoline, [0052]
4-(3′-fluorophenyl)amino-6,7-dimethoxyquinazoline, [0053]
4-(2′-fluorophenyl)amino-6,7-dimethoxyquinazoline, [0054]
4-(4′-trifluoromethylphenyl)amino-6,7-dimethoxyquinazoline, [0055]
4-(2′-trifluoromethylphenyl)amino-6,7-dimethoxyquinazoline, and [0056]
4-(3′,5′-bis-trifluoromethylphenyl)amino-6,7-dimethoxyquinazoline. [0057]
Compounds of the invention that are particularly useful for treating breast tumors include: [0058]
4-(3′-bromophenyl)amino-6,7-dimethoxyquinazoline, [0059]
4-(3′,5′-dibromo-4′-hydroxyphenyl)amino-6,7-dimethoxyquinazoline, [0060]
4-(3′-chloro-4′-hydroxyphenyl)amino-4,7-dimethoxyquinazoline, [0061]
4-(3′,5′-bis-trifluoromethylphenyl)amino-6,7-dimethoxyquinazoline, [0062]
4-(2′,3′,5′,6′-tetrafluoro-4′-bromophenyl)amino-6,7-dimethoxyquinazoline, [0063]
4-(4′-fluorophenyl)amino-6,7-dimethoxyquinazoline, [0064]
4-(3′-fluorophenyl)amino-6,7-dimethoxyquinazoline, and [0065]
4-(2′-fluorophenyl)amino-6,7-dimethoxyquinazoline. [0066]
Compositions [0067]
The compounds of the invention are useful as pharmaceutical compositions prepared with a therapeutically effective amount of a compound of [0068] claim 1 and a pharmaceutically acceptable carrier or diluent.
The quinazoline compounds of the invention can be formulated as pharmaceutical compositions and administered to a mammalian host, such as a human patient in a variety of forms adapted to the chosen route of administration, i.e., orally or parenterally, by intravenous, intramuscular, topical or subcutaneous routes. [0069]
Thus, quinazoline compounds of the invention may be systemically administered, e.g., orally, in combination with a pharmaceutically acceptable vehicle such as an inert diluent or an assimilable edible carrier, or by inhalation or insufflation. They may be enclosed in hard or soft shell gelatin capsules, may be compressed into tablets, or may be incorporated directly with the food of the patient's diet. For oral therapeutic administration, the quinazoline compounds may be combined with one or more excipients and used in the form of ingestible tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups, wafers, and the like. The quinazoline compounds may be combined with a fine inert powdered carrier and inhaled by the subject or insufflated. Such compositions and preparations should contain at least 0.1% quinazoline compounds. The percentage of the compositions and preparations may, of course, be varied and may conveniently be between about 2 to about 60% of the weight of a given unit dosage form. The amount of quinazoline compounds in such therapeutically useful compositions is such that an effective dosage level will be obtained. [0070]
The tablets, troches, pills, capsules, and the like may also contain the following: binders such as gum tragacanth, acacia, corn starch or gelatin; excipients such as dicalcium phosphate; a disintegrating agent such as corn starch, potato starch, alginic acid and the like; a lubricant such as magnesium stearate; and a sweetening agent such as sucrose, fructose, lactose or aspartame or a flavoring agent such as peppermnint, oil of wintergreen, or cherry flavoring may be added. When the unit dosage form is a capsule, it may contain, in addition to materials of the above type, a liquid carrier, such as a vegetable oil or a polyethylene glycol. Various other materials may be present as coatings or to otherwise modify the physical form of the solid unit dosage form. For instance, tablets, pills, or capsules may be coated with gelatin, wax, shellac or sugar and the like. A syrup or elixir may contain the active compound, sucrose or fructose as a sweetening agent, methyl and propylparabens as preservatives, a dye and flavoring such as cherry or orange flavor. Of course, any material used in preparing any unit dosage form should be pharmaceutically acceptable and substantially non-toxic in the amounts employed. In addition, the quinazoline compounds may be incorporated into sustained-release preparations and devices. [0071]
The quinazoline compounds may also be administered intravenously or intraperitoneally by infusion or injection. Solutions of the quinazoline compounds can be prepared in water, optionally mixed with a nontoxic surfactant. Dispersions can also be prepared in glycerol liquid polyethylene glycols, triacetin and mixtures thereof and in oils. Under ordinary conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms. [0072]
The pharmaceutical dosage forms suitable for injection or infusion can include sterile aqueous solutions or dispersions or sterile powders comprising the quinazoline compounds which are adapted for the extemporaneous preparation of sterile injectable or infusible solutions or dispersions, optionally encapsulated in liposomes. In all cases, the ultimate dosage form must be sterile, fluid and stable under the conditions of manufacture and storage. The liquid carrier or vehicle can be a solvent or liquid dispersion medium comprising, for example, water, ethanol, a polyol (for example, glycerol, propylene glycol, liquid polyethylene glycols, and the like), vegetable oils, nontoxic glyceryl esters, and suitable mixtures thereof. The proper fluidity can be maintained, for example, by the formation of liposomes, by the maintenance of the required particle size in the case of dispersions or by the use of surfactants. The prevention of the action of microorganisms can be brought about by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars, buffers or sodium chloride. Prolonged absorption of the injectable compositions can to be brought about by the use in the compositions of agents delaying absorption, for example, aluminum monostearate and gelatin. [0073]
Sterile injectable solutions are prepared by incorporating the quinazoline compounds in the required amount in the appropriate solvent with various of the other ingredients enumerated above, as required, followed by filter sterilization. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum drying and the freeze drying techniques, which yield a powder of the active ingredient plus any additional desired ingredient present in the previously sterile-filtered solutions. [0074]
For topical administration, the quinazoline compounds may be applied in pure form, i.e., when they are liquids. However, it will generally be desirable to administer them to the skin as compositions or formulations, in combination with a dermatologically acceptable carrier, which may be a solid or a liquid. [0075]
Useful solid carriers include finely divided solids such as talc, clay, microcrystalline cellulose, silica, alumina and the like. Other solid carriers include nontoxic polymeric nanoparticles or microparticles. Useful liquid carriers include water, alcohols or glycols or water-alcohol/glycol blends, in which the quinazoline compounds can be dissolved or dispersed at effective levels, optionally with the aid of non-toxic surfactants. Adjuvants such as fragrances and additional antimicrobial agents can be added to optimize the properties for a given use. The resultant liquid compositions can be applied from absorbent pads, used to impregnate bandages and other dressings, or sprayed onto the affected area using pumppe or aerosol sprayers. [0076]
Thickeners such as synthetic polymers, fatty acids, fatty acid salts and esters, fatty alcohols, modified celluloses or modified mineral materials can also be employed with liquid carriers to form spreadable pastes, gels, ointments, soaps, and the like, for application directly to the skin of the user. [0077]
Examples of useful dermatological compositions which can be used to deliver the quinazoline compounds to the skin are known to the art; for example, see Jacquet et al. (U.S. Pat. No. 4,608,392), Geria (U.S. Pat. No. 4,992,478), Smith et al. (U.S. Pat. No. 4,559,157) and Wortztman (U.S. Pat. No. 4,820,508). [0078]
Useful dosages of the compounds of formula I can be determined by comparing their in vitro activity, and in vivo activity in animal models. Methods for the extrapolation of effective dosages in mice, and other animals, to humans are known to the art; for example, see U.S. Pat. No. 4,938,949. [0079]
Generally, the concentration of the quinazoline compounds in a liquid composition, such as a lotion, will be from about 0.1-25 wt-%, preferably from about 0.5-10 wt-%. The concentration in a semi-solid or solid composition such as a gel or a powder will be about 0.1-5 wt-%, preferably about 0.5-2.5 wt-%. [0080]
The amount of the quinazoline compounds required for use in treatment will vary not only with the particular salt selected but also with the route of administration, the nature of the condition being treated and the age and condition of the patient and will be ultimately at the discretion of the attendant physician or clinician. [0081]
In general, however, a suitable dose will be in the range of from abut 0.5 to about 100 mg/kg, e.g., from about 10 to about 75 mg/kg of body weight per day, such as 3 to about 50 mg per kilogram body weight of the recipient per day, preferably in the range of 6 to 90 mg/kg/day, most preferably in the range of 15 to 60 mg/kg/day. [0082]
The quinazoline compounds are conveniently administered in unit dosage form; for example, containing 5 to 1000 mg, conveniently 10 to 750 mg, most conveniently, 50 to 500 mg of active ingredient per unit dosage form. [0083]
Ideally, the quinazoline compounds should be administered to achieve peak plasma concentrations of from about 0.5 to about 75 μM, preferably, about 1 to 50 μM, most preferably, about 2 to about 30 μM. This may be achieved, for example, by the intravenous injection of a 0.05 to 5% solution of the quinazoline compounds, optionally in saline, or orally administered as a bolus containing about 1-100 mg of the quinazoline compounds. Desirable blood levels may be maintained by continuous infusion to provide about 0.01-5.0 mg/kg/hr or by intermittent infusions containing about 0.4-15 mg/kg of the quinazoline compounds. [0084]
The quinazoline compounds may conveniently be presented in a single dose or as divided doses administered at appropriate intervals, for example, as two, three, four or more sub-doses per day. The sub-dose itself may be further divided, e.g., into a number of discrete loosely spaced administrations; such as multiple inhalations from an insulator or by application of a plurality of drops into the eye. [0085]
Targeting Quinazolines to Cells [0086]
In a preferred embodiment, the quinazoline compound is targeted to cells where treatment is desired, for example, to leukemia cells, to breast cells, or to other tumor cells. The compound is targeted to the desired cell by conjugation to a targeting moiety that specifically binds the desired cell, thereby directing administration of a conjugated molecule. Useful targeting moieties are ligands which specifically bind cell antigens or cell surface ligands, for example, antibodies against the B cell antigen, CD19 (such as B43) and the like. [0087]
To form the conjugates of the invention, targeting moieties are covalently bonded to sites on the quinazoline compound. The targeting moiety, which is often a polypeptide molecule, is bound to compounds of the invention at reactive sites, including NH[0088] ₂, SH, CHO, COOH, and the like. Specific linking agents are used to join the compounds. Preferred linking agents are chosen according to the reactive site to which the targeting moiety is to be attached.
Methods for selecting an appropriate linking agent and reactive site for attachment of the targeting moiety to the compound of the invention are known, and are described, for example, in Hermanson, et al., [0089] Bioconjugate Techniques, Academic Press, 1996; Hermanson, et al., Immobilized Affinity Ligand Techniques, Academic Press, 1992; and Pierce Catalog and Handbook, 1996, pp. T155-T201.
Administration of Quinazolines [0090]
According to the invention, quinazoline compounds may be administered prophylactically, i.e., prior to onset the pathological condition, or the quinazoline compounds may be administered after onset of the reaction, or at both times. [0091]

EXAMPLES

The invention may be further clarified by reference to the following Examples, which serve to exemplify some of the preferred embodiments, and not to limit the invention in any way. [0092]

Example 1

Synthesis of Quinazoline Derivatives

All chemicals were purchased from the Aldrich Chemical Company, Milwaukee, Wis., and were used directly for synthesis. Anhydrous solvents such as acetonitrile, methanol, ethanol, ethyl acetate, tetrahydrofuran, chloroform, and methylene chloride were obtained from Aldrich as sure seal bottles under nitrogen and were transferred to reaction vessels by cannulation. All reactions were carried out under a nitrogen atmosphere. [0093]
The key starting material, 4-chloro-6,7-dimethoxyquinazoline, was prepared according to published procedures (Nomoto, et al., 1990, [0094] Chem. Pharm. Bull., 38:1591-1595; Thomas, C. L., 1970, IN:Catalytic Processes and Proven Catalysts, Academic Press, New York, N.Y.) as outlined below in Scheme 1. Specifically, 4,5-dimethoxy-2-nitrobenzoic acid (compound 1) was treated with thionyl chloride to form acid chloride, followed by reacting with ammonia to yield 4,5-dimethoxy-2-nitrobenzamide (compound 2). Compound 2 was reduced with sodium borohydride in the presence of catalytic amounts of copper sulphate to give 4,5-dimethoxy-2-aminobenzamide (compound 3), which was directly refluxed with formic acid to yield 6,7-dimethoxyquinazoline-4(3H)-one (compound 4). Compound 4 was refluxed with phosphorus oxytrichloride to give 4-chloro-6,7-dimethoxyquinazoline (compound 5) in good yield.
Substituted quinazoline derivatives were prepared by the condensation of 4-chloro-6,7-dimethoxyquinazoline with substituted anilines as outlined below in Scheme 2: [0095]
Specifically, a mixture of 4-chloro-6,7-dimethoxyquinazoline (448 mg, 2 mmols) and the substituted aniline (2.5 mmols) in EtOH (20 ml) was heated to reflux. After refluxing for 4-24 hours, an excess amount of Et[0096] ₃N was added, and the solvent was concentrated to give the crude product which was recrystalized from DMF.
As discussed above, the novel hydroxy-substituted quinazoline derivatives of the invention were created by reacting the appropriate substituted anilines with the key starting material, 4-chloro-6,7-dimethoxyquinazoline. [0097]
Physical Characteristics: [0098]
Melting points are uncorrected. [0099] ¹H NMR spectra were recorded using a Varian Mercury 300 spectrometer in DMSO-d₆or CDCl₃. Chemical shifts are reported in parts per million (ppm) with tetramethylsilane (TMS) as an internal standard at zero ppm. Coupling constants (3) are given in hertz and the abbreviations s, d, t, q, and m refer to singlet, doublet, triplet, quartet and multiplet, respectively. Infrared spectra were recorded on a Nicolet PROTEGE 460-IR spectrometer. Mass spectroscopy data were recorded on a FINNIGAN MAT 95, VG 7070E-HF G.C. system with an HP 5973 Mass Selection Detector. UV spectra were recorded on BECKMAN DU 7400 and using MEOH as the solvent. TLC was performed on a precoated silica gel plate (Silica Gel KGF; Whitman Inc). Silica gel (200-400 mesh, Whitman Inc.) was used for all column chromatography separations. All chemicals were reagent grade and were purchased from Aldrich Chemical Company (Milwaukee, Wis.) or Sigma Chemical Company (St. Louis, Mo.).

Example 2

Bromine Substituted Quinazoline Compounds

Bromine substituted quinazoline derivatives were synthesized and characterized as discussed above in Example 1. The structures and physical data are shown below:

Bromine Substituted Quinazoline Compounds

No	Name	Structure	Formula	MW


1	P-79	C₁₆H₁₄BrN₃O₂	360

2	P-88	C₁₇H₁₄BrN₃O₄	404

3	P-97	C₁₆H₁₃Br₂N₃O₃	455

4	P-111	C₁₇H₁₆BrN₃O₂	374

5	P-112	C₁₆H₁₃Br₂N₃O₂	439

6	P-154	C₁₆H₁₄BrN₃O₃	376

7	P-160	C₂₃H₁₈BrN₃O₂	448

8	P-164	C₁₇H₁₃BrN₂O₃	373

9	P-190	C₁₇H₁₆BrN₃O₃	389

10	P-210	C₁₇H₁₅Br₂N₃O₂	453

11	P-211	C₁₇H₁₅Br₂N₃O₂	453

12	P-212	C₁₇H₁₅Br₂N₃O₂	453

13	P-214	C₁₆H₁₃BrFN₃O₂	378

14	P-222	C₁₆H₁₂Br₃N₃O₂	518

15	P-234	C₁₇H₁₇N₃O₂	295

16	P-241	C₁₇H₁₅Br₂N₃O₂	453

17	P-258	C₁₆H₁₅N₃O₂	281

18	P-260	C₁₆H₁₄BrN₃O₂	360

19	P-261	C₁₆H₁₄BrN₃O₂	360

20	P-262	C₁₆H₁₃Br₂N₃O₂	439

21	P-263	C₁₆H₁₃Br₂N₃O₂	439

4-(3′-Bromophenyl)-amino-6,7-dimethoxyquinazoline (HI-P79)

Yield 84.17%; m.p. 246.0-249.0° C. ¹H NMR(DMSO-d₆): δ 10.42(br, s, 1H, NH), 8.68(s, 1H, 2-H),

8.07-7.36(m, 5H, 5,2′,4′,5′,6′-H), 7.24(s, 1H, 8H), 3.98(s, 3H, —OCH₃), 3.73(s, 3H, —OCH₃);

IR(KBr)ν_max: 3409, 2836, 1632, 1512, 1443, 1243, 1068 cm⁻¹;

GC/MS m/z 361(M⁺ + 1, 61.8), 360(M⁺, 100.0), 359(M⁺ − 1, 63.5), 344(11.3), 222(10.9), 140(13.7).

Anal. (C₁₆H₁₄BrN₃O₂HCl) C, H, N.

4-(4′-Bromo-2′-caboxylphenyl)-amino-6,7-dimethoxyquinazoline (HI-P88)

Yield 92.82%; m.p. > 300.0° C. ¹H NMR(DMSO-d₆+ CF₃CO₂H): δ 9.95(d, 1H),

8.74(d, 1H, Ar—H), 8.30, 8.28(2d, 2H), 7.95(d, 1H), 7.83(s, 1H), 4.21(s, 3H, —OCH₃),

4.15(s, 3H, —OCH₃). UV(MeOH): 205, 229.0 nm. IR(KBr)ν_max: 3444(br), 2737, 1592, 1504, 1443,

1273, 1070 cm⁻¹. GC/MS m/z 388(M⁺ + 1 —OH, 18.08), 387(M⁺ —OH, 100.00),

386(M⁺ − 1 —OH, 30.84), 385(97.52), 299(4.78). Anal. (C₁₆H₁₄BrN₃O₂HCl) C, H, N.

4-(3′,5′-Dibromo-4′-hydroxylphenyl)-amino-6,7-dimethoxyquinazoline (HI-P97).

Yield 72.80%; m.p. > 300.0° C. ¹H NMR(DMSO-d₆): δ 9.71(s, 1H, —NH), 9.39(s, 1H, —OH),

8.48(s, 1H, 2-H), 8.07(s, 2H, 2′,6′-H), 7.76(s, 1H, 5-H), 7.17(s, 1H, 8-H), 3.94(s, 3H, —OCH₃),

3.91(s, 3H, —OCH₃). UV(MeOH): 208.0, 210.0, 245.0, 320.0 nm; IR(KBr)ν_max:

3504(br), 3419, 2868, 1627, 1512, 1425, 1250, 1155 cm⁻¹; GC/MS m/z 456(M⁺ + 1, 54.40),

455(M⁺, 100.00), 454(M⁺ − 1, 78.01), 439(M⁺ —OH, 7.96), 376(M⁺ + 1 —Br, 9.76),

375(M⁺ —Br, 10.91), 360(5.23). Anal. (C₁₆H₁₃Br₂N₃O₃) C, H, N.

4-(3′-Bromo-4′-methylphenyl)-amino-6,7-dimethoxyquinazoline (HI-P111):

Yield 82.22%; m.p. 225.0-228° C. ¹H NMR(DMSO-d₆): δ 10.23(s, 1H, —NH), 8.62(s, 1H, 2-H),

8.06(d, 1H, J_2′,6′=2.1 Hz, 2′-H), 7.89(s, 1H, 5-H), 7.71(dd, 1H, J_5′,6′=8.7 Hz, J_2′,6′=2.1 Hz, 6′-H),

7.37(d, 1H, J_5′,6′=8.7 Hz, 5′-H), 7.21(s, 1H, 8-H), 3.96(s, 3H, —OCH₃), 3.93(s, 3H, —OCH₃).

UV(MeOH): 204.0, 228.0, 255.0, 320.0 nm. IR(KBr)ν_max: 3431, 3248, 2835,

1633, 1517, 1441, 1281, 1155 cm⁻¹. GC/MS m/z 375(M⁺ + 1, 76.76), 374(M⁺, 100.00),

373(M⁺ − 1, 76.91), 358(M⁺ + 1 —OH, 11.15), 357(1.42), 356(6.31).

Anal. (C₁₇H₁₆BrN₃O₂.HCl) C, H, N.

4-(2′,5′-Dibromophenyl)-amino-6,7-dimethoxyquinazoline (HI-P112):

Yield 70.05%; m.p. > 300.0° C. ¹H NMR(DMSO-d₆): δ 11.51(s, 1H, —NH), 8.76(s, 1H, 2-H),

8.21(s, 1H, 5-H), 7.81(d, 1H, J_4′,6′=2.4 Hz, 6′-H), 7.75(d, 1H, J_3′,4′=8.7 Hz, 3′-H),

7.55(dd, 1H, J_4′,6′=2.4 Hz, J_3′,4′=8.7 Hz, 4′-H), 7.33(s, 1H, 8-H), 3.98(s, 3H, —OCH₃), 3.97(s, 3H,

—OCH₃). UV(MeOH): 208.0, 238.0, 330.0 nm. IR(KBr)ν_max: 3444, 2836, 1628, 1510, 1431, 1277,

1070 cm⁻¹. GC/MS m/z 440(M⁺ + 1, 10.12), 439(M⁺, 7.0), 438(M⁺ − 1, 3.63),

360(M⁺ + 1 —Br, 99.42), 359(M⁺ —Br, 20.45), 358(M⁺ − 1 —Br, 100.00),

343(20.80), 299(8.62). Anal. (C₁₆H₁₃Br₂N₃O₂.HCl) C, H, N.

4-[(3′-Bromo-9′-fluorenone)-2′]-amino-6,7-dimethoxyquinazoline (HI-P119):

Yield 75.23%; m.p. 255.0-257.0° C. ¹H NMR(DMSO-d₆): δ 8.77(s, 1H, —NH), 8.33(s, 1H, 2-H).

7.89(s, 1H, 5-H), 7.40(s, 1H, 8-H), 7.74-7.26(m, 6H, Ar—H), 4.12(s, 3H, —OCH₃),

4.11(s, 3H, —OCH₃). UV(MeOH): 205, 229.0, 251.0, 320.0 nm.

IR(KBr)ν_max: 3444, 2836, 1628, 1510, 1431, 1277, 1070 cm⁻¹.

GC/MS m/z 464(M⁺ + 2, 40.81), 463(M⁺ + 1, 7.56), 462(M⁺, 27.26), 384(M⁺ + 2 —Br, 69.56),

383(M⁺ + 1 —Br, 35.50), 382(M⁺ —Br, 100.00), 352(10.85),

206(26.73), 191(11.31). Anal. (C₂₃H₁₆BrN₃O₃HCl) C, H, N.

4-(2′,3′,5′,6′-Tetrafluoro-4′-bromolphenyl)amino-6,7-dime-thoxyquinazoline (HI-P144:

Yield 78.24%; m.p. 180.0-182.0° C. ¹H NMR(DMSO-d₆): δ 7.78(s, 1H, 2-H), 7.53(s, 1H, 5-H),

6.79(s, 1H, 8-H), 3.81(s, 3H, —OCH₃), 3.3.79(s, 3H, —OCH₃).

Anal(C₁₆H₁₀BrF₄N₃O₂.HCl) C, H, N.

4-(3′-Bromo-4′-hydroxylphenyl)-amino-6,7-dimethoxyquinazoline (HI-P154):

Yield 89.90%; m.p. 233.0-233.5° C. ¹H NMR(DMSO-d₆): δ 10.08(s, 1H, —NH), 9.38(s, 1H, —OH),

8.40(s, 1H, 2-H), 7.89(d, 1H, J_2′,6′=2.7 Hz, 2′-H), 7.75(s, 1H, 5-H), 7.55(dd, 1H, J_5′,6′=9.0 Hz,

J_2′,6′=2.7 Hz, 6′-H), 7.14(s, 1H, 8-H), 6.97(d, 1H, J_5′,6′=9.0 Hz, 5′-H),

3.92(s, 3H, —OCH₃), 3.90(s, 3H, —OCH₃). UV(MeOH): 203.0, 222.0, 250.0, 335.0 nm.

IR(KBr)ν_max: 3431(br), 2841, 1624, 1498, 1423, 1244 cm⁻¹. GC/MS m/z 378(M⁺ + 2, 90.68),

377(M⁺ + 1, 37.49), 376(M⁺, 100.00), 360(M⁺, 3.63), 298(18.86), 282(6.65).

Anal. (C₁₆H₁₄BrN₃O₃.HCl) C, H, N.

4-[(7′-Bromofluorene)-2′]-amino-6,7-dimethoxyquinazoline (HI-P160):

Yield 73.21%; m.p. 254.0-256.0° C. ¹H NMR(DMSO-d₆): δ 9.69(br, s, 1H, —NH), 8.52(s, 1H, 2-H),

8.12-7.20(m, 9H, 5, 8, 1′, 3′, 4′, 5′, 6′, 8′, 9′-H), 3.99(s, 3H, —OCH₃), 3.94(s, 3H, —OCH₃).

UV(MeOH): 208.0, 223.0, 348.0 nm. IR(KBr)ν_max: 3421, 2820, 1624, 1516, 1431, 1294, 1223 cm⁻¹.

GC/MS m/z 450(M⁺ + 2, 100), 449(M⁺ + 1, 35), 448(M⁺, 95), 311(25).

Anal. (C₂₃H₁₈BrN₃O₂.HCl) C, H, N.

4-(3′-Bromobenzoyl)-6,7-dimethoxyquinazoline (HI-P164).

Yield 81.20%, m.p. 258.0-263.0° C. ¹H NMR(DMSO-d₆): δ 9.25(s, 1H, 2-H), 8.14(s, 1H, 5-H),

7.92-7.43(m, 4H, 2′,4′,5′,6′-H), 7.40(s, 1H, 8-H), 4.11(s, 3H, —OCH₃), 4.00(s, 3H, —OCH₃).

UV(MeOH): 203.0, 220.0, 238.0 nm. IR(KBr)ν_max: 3432, 1664, 1504, 1431, 1230 cm⁻¹.

GC/MS m/z 374(M⁺ + 1, 48.96), 373(M⁺, 34.93), 372(M⁺ − 1, 47.67), 357(58.74), 343(100.00),

293(M⁺ —Br, 31.48), 189(26.27). Anal. (C₁₇H₁₃BrN₂O₃) C, H, Br, N.

4-(4′-Bromo-6′-hydroxymethylphenyl)-amino-6,7-dimethoxyquinazoline (HI-P190):

Yield 73.08%; m.p. 222.0-223.0° C. ¹H NMR(DMSO-d₆): δ 11.30(s, 1H, —OH), 8.22(s, 1H, —NH),

7.77-7.23(m, 5H, 5,8,2′,3′,5′-H), 4.49(s, 2H, PhCH₂—H), 4.01(s, 3H, —OCH₃),

3.90(s, 3H, —OCH₃). UV(MeOH): 207.0, 250.0, 332.0 nm. IR(KBr)ν_max: 3446, 2829, 2752,

1652, 1560, 1471, 1365, 1280 cm⁻¹. GC/MS m/z 391(M⁺ + 1, 29.33), 389(M⁺, 29.82),

360(M⁺ —CH₂OH, 50.76), 358(52.39), 311(18.33), 280(43.20), 206(62.80), 191(100.00).

Anal. (C₁₇H₁₆BrN₃O₃.HCl) C, H, N.

4-(2′,3′-Dibromo-4′-methylphenyl)-amino-6,7-dimethoxyquinazoline (HI-P210):

Yield 81.24%, mp 233.0-236.0° C., ¹H NMR(DMSO-d₆): δ 8.55(s, 1H, —NH), 8.08(s, 1H, 2-H),

7.33-7.17(m, 4H, 5,8,5′,6′-H), 3.89(s, 6H, —OCH₃), 2.35(s, 3H, —CH₃).

UV(MeOH): 207.0, 232.0, 247.0, 330.0 nm. IR(KBr)ν_max: 3448, 2840, 1629, 1580, 1525, 1420,

1281 cm⁻¹. GC/MS m/z 454(M⁺ + 1, 4.45) , 453(M⁺, 11.31), 452(M⁺ − 1, 4.45),

375(20.36), 374(97.59), 373( 23.55), 372(100.00), 358(19.61), 356(18.43).

Anal. (C₁₇H₁₅Br₂N₃O₂.HCl) C, H, N.

4-(2′,5′-Dibromo-4′-methylphenyl)-amino-6,7-dimethoxyquinazoline (HI-P211):

Yield 83.50%; m.p. 282.0-284.0° C. ¹H NMR(DMSO-d₆): δ 11.30(s, 1H, —NH), 8.58(s, 1H, 2-H),

8.00(s, 1H, 5-H), 7.65(s, 1H, 6′-H), 7.60(s, 1H, 3′-H), 7.13(s, 1H, 8-H), 3.79(s, 3H, —OCH₃),

3.78(s, 3H, —OCH₃), 2.29(s, 3H, —CH₃). UV(MeOH): 207.0, 239.0, 330.0 nm.

IR(KBr)ν_max: 3442, 2620, 1631, 1580, 1514, 1380, 1280 cm⁻¹. GC/MS m/z 454(M⁺ + 1, 5.86),

453(M⁺, 16.16), 452(M⁺ − 1, 5.35), 374(92.12), 373(23.66), 372(100.00), 358(17.68),

356(17.35). Anal. (C₁₇H₁₅Br₂N₃O₂.HCl) C, H, N.

4-(3′,5′-Dibromo-4′-methylphenyl)-amino-6,7-dimethoxyquinazoline (HI-P212):

Yield 83.47%; m.p. 275.0-279.0° C. ¹H NMR(DMSO-d₆): δ 11.30(s, 1H, —NH), 8.58(s, 1H, 2-H),

8.35(s, 1H, 5-H), 7.24(s, 2H, 2′,6′-H), 7.13(s, 1H, 8-H), 3.91(s, 3H, —OCH₃), 3.88(s, 3H, —OCH₃),

2.31(s, 3H, —CH₃). UV(MeOH): 237.0, 307.0, 319.0 nm.

IR(KBr)ν_max: 3471, 3434, 2640, 1633, 1580, 1504, 1420, 1281 cm⁻¹.

GC/MS m/z 454(M⁺ + 1, 5.34), 453(M⁺, 16.05), 452(M⁺ − 1, 5.87), 374(99.02), 373(26.20),

372(100.00), 358(20.39), 356(19.98), 32(8.29), 314(8.49), 206(19.02).

Anal. (C₁₇H₁₅Br₂N₃O₂HCl) C, H, N.

4-(2′-Fluoro-4′-bromophenyl)-amino-6,7-dimethoxyquinazoline (HI-P214):

Yield 77.21%; m.p. 243.0-245.0° C. ¹H NMR(DMSO-d₆): δ 8.57(s, 1H, 2-H), 7.91(s, 1H, 5-H),

7.57(d, 1H, 3′-H), 7.34(m, 2H, 5′,6′-H), 7.07(s, 1H, 8-H), 3.78(s, 3H, —OCH₃),

3.77(s, 3H, —OCH₃). UV(MeOH): 204.0, 215.0, 250.0, 330.0 nm. IR(KBr)ν_max: 3431, 2629, 1633,

1580, 1511, 1420, 1278 cm⁻¹. GC/MS m/z 379(M⁺ + 1, 34.39), 378(M⁺, 21.33),

377(M⁺ − 1, 39.08), 360(62.05), 359(31.58), 358(62.57), 357(19.81), 299(19.31),

298(100.00), 282(17.88), 240(28.76). Anal. (C₁₆H₁₃BrFN₃O₂HCl) C, H, N.

4-(2′,4′,6′-Tribromophenyl)amino-6,7-dimethoxyquinazoline (HI-P222):

Yield 54.86%; m.p. 250.0-255.0° C. ¹H NMR(DMSO-d₆): δ 8.00(s, 1H, 2-H), 7.89(s, 2H, 3′, 5′-H),

7.74(s, 1H, 5-H), 7.01(s, 1H, 8-H), 3.87(s, 3H, —OCH₃), 3.86(s, 3H, —OCH₃).

UV(MeOH): 209.0, 236.0, 333.0 nm. IR(KBr)ν_max: 3417, 2838, 1625, 1514, 1429, 1276, 1073 cm⁻¹.

GC/MS m/z 519(M⁺ + 1, 18.12), 518(M⁺, 17.30), 517(M⁺ − 1, 16.63),

439(M⁺ + 1-Br, 99.42), 438(M⁺ − Br, 95.45), 437(M⁺ − 1 —Br, 100.00),

359(20.80), 358(18.62), 357(19.32), 281(88.98), 207(15.42).

Anal. (C₁₆H₁₂Br₃N₃O₂HCl) C, H, N.

4-(2′,6′-Dibromo-4′-methylphenyl)-amino-6,7-dimethoxyquinazoline (HI-P241):

Yield 79.47%, m.p. 235.0-237.0° C. ¹H NMR(DMSO-d₆): δ 9.77(s, 1H, —HN), 8.20(s, 1H, 2-H),

7.87(s, 1H, 8-H), 7.61(s, 2H, 3′,5′-H), 7.15(s, 1H, 5-H), 3.93(s, 6H, —OCH₃).

UV(MeOH): 208.0, 245.0, 318.0, 339.0 nm. IR(KBr)ν_max: 3241, 2839, 2783, 1635, 1580, 1514,

1420, 1360, 1281 cm⁻¹. GC/MS m/z 454(M⁺ + 1, 7.86), 453(M⁺, 56.16), 452(M⁺ − 1, 15.30),

374(95.12), 373(18.66), 372(100.00), 358(29.64), 356(19.36). Anal. (C₁₇H₁₅Br₂N₃O₂HCl) C, H, N.

4-(4′-Bromophenyl)-amino-6,7-dimethoxyquinazoline (HI-P260):

Yield 75.28%. m.p. 270.0-272.0° C. ¹H NMR(DMSO-d₆): δ 11.30(s, 1H, —NH), 8.85(s, 1H, 2-H),

8.27(s, 1H, 5-H), 7.70(s, 4H, 2′,3′,5′,6′-H), 7.32(s, 1H, 8H), 4.02(s, 3H, —OCH₃).

4.00(s, 3H, —OCH₃). UV(MeOH): 204.0, 218.0, 252.0, 335.0 nm.

IR(KBr)ν_max: 3431, 3034, 2636, 1635, 1589, 1514, 1435, 1284 cm⁻¹.

GC/MS m/z 361(M⁺ + 1, 74.00), 360(M⁺, 100.00),

359(M⁺ − 1, 72.00), 358(M⁺ − 2, 95.00), 329(3.20), 301(13.0), 281(21.0), 207(38.0).

Anal. (C₁₆H₁₄BrN₃O₂.HCl) C, H, N.

4-(2′-Bromophenyl)-amino-6,7-dimethoxyquinazoline (HI-P261):

Yield 71.94%; m.p. 241.0-243.0° C. ¹H NMR(DMSO-d₆) δ 11.67(d, 1H, —NH), 8.79(s, 1H, 2-H),

8.32(s, 1H, 5-H), 7.86-7.38(m, 4H, 3′,4′,5′,6′-H), 7.40(s, 1H, 8H), 4.01(s, 6H, —OCH₃).

UV(MeOH): 204.0, 226.0, 248.0, 330.0 nm. IR(KBr)ν_max: 3454, 3032, 2638, 1630, 1589, 1514,

1430, 1281 cm⁻¹. GC/MS m/z 361(M⁺ + 1, 7.00), 360(M⁺, 5.00),

359(M⁺ − 1, 6.00), 358(M⁺ − 2, 5.00), 301(13.0), 281(21.0), 280(100.00), 207(25.00).

Anal (C₁₆H₁₄BrN₃O₂.HCl) C, H, N.

4-(2′,6′-Dibromophenyl)-amino-6,7-dimethoxyquinazoline (HI-P262):

Yield 69.45%, mp 243.0-246.0° C., ¹H NMR(DMSO-d₆): δ 11.91(d, 1H, —NH), 8.80(s, 1H, 2-H),

8.43(s, 1H, 5-H), 7.86(d, 2H, J=8.4 Hz, 3′,5′-H), 7.49(s, 1H, 8H), 7.35(t, 1H, J=8.4 Hz, 4′-H),

4.02(s, 3H, —OCH₃), 4.01(s, 3H, —OCH₃). UV(MeOH): 208.0, 227.0, 245.0, 330.0 nm.

IR(KBr)ν_max: 3454, 3032, 2638, 1630, 1589, 1514, 1430, 1281 cm⁻¹.

4-(2′,4′-Dibromophenyl)-amino-6,7-dimethoxyquinazoline (HI-P263):

Yield 70.62%; m.p. 257.0-262.0° C. ¹H NMR(DMSO-d₆): δ 11.91(d, 1H, —NH), 8.79(s, 1H, 2-H),

8.21(s, 1H, 5-H), 8.12-7.51(m, 3H, 3′,5′,6′-H), 7.35(s, 1H, 8-H), 4.01(s, 3H, —OCH₃),

3.99(s, 3H, —OCH₃). UV(MeOH): 208.0, 210.0, 248.0, 330.0 nm.

IR(KBr)ν_max: 3458, 3028, 2641, 1633, 1594, 1511, 1435, 1277 cm⁻¹.

Example 3

Chlorine Substituted Quinazoline Compounds

Chlorine substituted quinazoline derivatives were synthesized and characterized as

discussed above in Example 1. The structures and physical data are shown below:

No	Name	Structure	Formula	MW


1	P-87	C₁₆H₁₄ClN₃O₂	316

2	P-93	C₁₆H₁₄ClN₃O₃	331

3	P-189	C₁₆H₁₃Cl₂N₃O₃	365

4	P-197	C₁₆H₁₄ClN₃O₃	331

5	P-268	C₁₆H₁₄ClN₃O₂	316

6	P-269	C₁₆H₁₄ClN₃O₂	316

7	P-278	C₁₆H₁₄ClN₃O₃	331

8	P-415	C₂₀H₁₆ClN₃O₂	365

4-(3′-Chlorophenyl)-amino-6,7-dimethoxyquinazoline (HI-P87). Yield 76.98%;

m.p. 242.0-245.0° C. ¹H NMR(DMSO-d₆): δ 10.47(br, s, 1H, NH), 8.69(s, 1H, 2-H),

8.06(s, 1H, 5-H), 7.95-7.23(m, 4H, 2′,4′,5′,6′-H), 7.24(s, 1H, 8-H), 3.98(s, eH, —OCH₃),

3.35(s, 3H, 0OCH₃). UV(MeOH): 228.0, 251.0, 332.0 nm. IR(KBr)ν_max: 3406, 2839, 1632,

1516, 1443, 1278, 1068 cm⁻¹. GC/MS m/z 316(M⁺ − 1, 68.34), 314(M⁺ − 2,

100.00, 344(11.34), 222(4.35), 140(9.86). Found: C, 54.62; H, 4.68; N, 11.93; Cl, 19.23.

C₁₆H₁₄ClN₃O₂.HCl requires: C, 54.70; H, 4.28; N, 11.96; Cl, 19.96%.

4-(c′-Chloroo-6′-hydroxyphenyl)amino-6,7-dimethoxyquinazoline (HI-P93)

Yield 83.08%; m.p. 295.0° C.(dec). ¹H NMR9DMSO-d₆: δ 10.14(s, 1H, —OH),

8.37(s, 1H, 2-H), 7.78(s, 1H, 5H), 7.57(d, 1H, J_2′,4′=2.4 Hz, 2′-H), ), 7.16(s, 1H, 8-H),

7.07(dd, 1H, J_2′,4′=2.4 Hz, J_4′,5′=8.7 Hz, 4′-H),

6.92(d, 1H, J_4′,5′=8.7 Hz, 5′-H), 3.93(s, 3H, —OCH₃).

3.92(s, 3H, —OCH₃). UV(MeOH): 205, 229.0, 251.0, 320.0 nm. IR(KBr)ν_max: 3500(br),

3430, 2835, 1622, 1512, 1432, 1259 cm⁻¹. GC/MS m/z 333(M⁺ + 2, 13.41),

332(M⁺ + 1, 9.73, 331(M⁺, 39.47), 314(M⁺ —OH, 100.00), 298(7.64).

Found: C, 52.25; H, 4.07; N, 11.39. C₁₆H₁₄ClN₃O₃.HCl

requires: C, 52.32; H, 4.09; N, 11.44%.

4-(4′-Hydroxyl-3′,5′-dichlorophenyl)amino-6,7-dimethoxyquinazoline (HI-P189)

Yield 79.45%; m.p. 293.0-295.0° C. ¹H NMR-DMSO-d₆): δ 11.32(s, 1H, —NH),

10.34(s, 1H, —OH), 8.87(s, 1H, 2-H), 8.29(s, 1H, 5-H), 7.90(s, 2H, 2′,6′-H),

7.32(s, 1H, 8-H), 4.01(s, 3H, —OCH₃), 3.99(s, 3H, —OCH₃).

UV(MeOH): 213.0, 232.0, 250.0, 335.0 nm. IR(KBr)ν_max: 3479, 2564, 1641, 1579, 1429,

1282, 1147 cm⁻¹. GC/MS m/z 367(M⁺ = 2, 66.57), 366(M⁺=1, 75.91),

365(M⁺, 100.00), 364(M⁺ − 194.08), 349(M⁺ —OH, 11.16).

Anal. (C₁₆H₁₃Cl₂N₃O₃) C, H, N.

Found: C, 48.93; H, 4.51; N, 10.00. C_17oH₁₇Cl₂N₃O₃.Hcl

requires: C, 48.80; H, 4.31; N, 10.04.%.

4-(3′-Chloro-4′-hydroxylphenyl)-amino-6,7-dimethoxyquinazoline (HI-P197).

Yield 84.14%; m.p. 245.0° C.(dec). ¹H NMR(DMSO-d₆): δ 10.00(s, 1H, —NH),

9.37(s, 1H, —OH), 8.41(s, 1H, 2-H), 7.78(s, 1H, 5-H), 7.49(d, 1H, J_2′,5′=2.7 Hz, 2′-H),

7.55(dd, 1H, J_5′,6′=9.0 Hz, J_2′,6′=2.7 Hz, 6′-H), 7.16(s, 1H, 8-H),

6.97(d, 1H, J_5′,6′=9.0 Hz, 5′H), 3.93(s, 3H, —OCH₃), 3.91(s, 3H, —OCH₃).

UV(MeOH): 209.0, 224.0, 249.0, 330.0 nm. IR(KBr)ν_max: 3448, 2842, 1623, 1506,

1423, 1241 cm⁻¹. GC/MX m/z: 341(M⁺, 100.00), 326(M⁺ —CH₃, 98.50),

310(M⁺ —OCH₃, 12.5), 295(9.0.), 189(13.5), 155(13.8).

Found: C, 521.35; H, 4.16; Cl, 19.15; N, 11.39. C₁₆H₁₄ClN₃O₃.HCl

requires: C, 52.32; H, 4.09; Cl, 19.07; N, 11.44%.

4-(2′-Chlorophenyl)-amino-6,7-dimethoxyquinazoline (HI-P268)

Yield 87.28%; m.p. 247.0-279.5° C. ¹H NMR(DMSO-d₆): δ 11.71(s, 1H, —NH),

8.78(s, 1H, 2-H), 8.33(s, 1H, 5-H), 7.67(s, 1H, 8H), 7.68-7.42(m, 4H, 3′,4,5,6′-H),

4.00(s, 3H, —OCH₃), 3.99(s, 3H, —OCH₃). UV(MeOH): 213.0, 234.0, 251.0, 331.0 nm.

IR(KBr)ν_max: 3479, 2566, 1643, 1577, 1429, 1282, 1147 cm⁻¹.

GC/MX m/z 317(M⁺ + 1, 6.60), 316(M⁺, 6.60), 315(M⁺ − 1, 18.52),

314(M⁺ − 2, 11.11), 281(21.22), 280(M⁺ —Cl, 100.00), 264(29.62).

Found: C, 54.51; H, 4.41; N, 11.81. C₁₆H₁₄ClN₃O₂.HCl

requires: C, 54.45; H, 4.26; N, 11.93%.

4-(4′-Chlorophenyl)-amino-6,7-dimethoxyquinazoline (HI-P269)

Yield 94.94%. m.p. 248.0-250.0° C. ¹H NMR(DMSO-d₆): δ 11.62(s, 1H, —NH),

8.85(s, 1H, 2-H), 8.42(s, 1H, 5-H), 7.88(d, 2H, J=8.7 Hz, 3′, 5′, —H),

7.54(d, 2H, J=8.7 Hz, 2′, 6′, —H), 7.38(s, 1H, 8-H0, 4.02(s, 3H, —OCH₃),

3.99(s, 3H, —OCH₃). UV(MeOH): 215.0, 230.0, 253.0, mn.

IR(KBr)ν_max: 3477, 2563, 1640, 1578 cm⁻¹.

GC/MX m/z 317(M⁺ + 1, 18.18), 316(M⁺, 29.55),

315(M⁺ − 1, 48.85), 314(M⁺ − 2, 61.36), 281(32., 95), 207(100.00).

Found: C, 54.65; H, 4.38; N, 11.92. C₁₆H₁₄ClN₃O₂.HCl

requires: C, 54.55; H, 4.26; N, 11.93%.

4-(4′-Hydroxyl-2′-chlorophenyl)-amino-6,7-dimethoxy-quinazoline (HI-P278)

Yield 81.44%; m.p. 245.0-247.0° C. ¹H NMR(DMSO-d₆): δ 11.39(s, 1H, —NH),

10.30(s, 1H, —OH), 8.75(s, 1H, 2-H), 8.24(s, 1H, 5-H), 7.38-6.85(m, 3H, 3′,5′,6′-H),

7.37(s, 1H, 8H), 3.98(s, 3H, —OCH₃), 3.96(s, 3H, —OCH₃). UV(MeOH): 222.0,

234.0, 239.0, 245.0, 254.0, 348.0 nm. IR(KBr)ν_max: 3448, 3242, 3144, 3025, 2917,

2834, 1638, 1591, 1514, 1437, 1365, 1277, 1209 cm⁻¹. GC/MS m/z: 332(M⁺ + 1, 5.00),

331(M⁺, 17.00), 330(M⁺ − 1, 5.00), 297(17.00), 296(100.00), 281(18.00),

280(29.00), 253(9.00). Found: C, 52.17; 11, 4.06; N, 11.32. C₁₆H₁₄ClN₃O₃.HCl

requires: C, 52.32; H, 4.01; N, 11.44%.

4-(4′-Chloronaphthy-1′)-amino-6,7-dimethoxyquinazoline (HI-P415)

Yield, 85.07%. m.p. 245.0-248.0° C. ¹H NMR(DMSO-d₆): δ 11.91(s, 1H, —NH),

8.66(s, 1H, 2-H), 8.45(s, 1H, 5-H), 8.30-7.62(m, 6H, 2′,3′,5′,6′,7′,8′-H), 7.38(s, 1H, 8-H),

4.03(s, 3H, —OCH₃), 4.01(s, 3H, —OCH₃). UV(MeOH): 211.0, 233.0, 250.0, mn.

IR(KBr)ν_max: 3481, 2567, 1645, 1579 cm⁻¹.

Found: C, 59.32; H, 4.27; N, 10.24. C₂₀H₁₆ClN₃O₂.HCl

requires: C, 59.70; 11, 4.23; N, 10.48%.

Example 4

Iodine Substituted Quinazoline Compounds

Iodine substituted quinazoline derivatives were synthesized as discussed above in Example 1, and analyzed. The structures and physical data are shown below:

Iodine Substituted Quinazoline Compounds

No	Name	Structure	Formula	MW


1	P-270	C₁₆H₁₄IN₃O₂	407

2	P-271	C₁₆H₁₄IN₃O₂	407

3	P-300	C₁₆H₁₄IN₃O₂	407

4	P-294	C₁₆H₁₃I₂N₃O₃	549

5	P-299	C₁₆H₁₄IN₃O₃	423

4-(2′-Iodophenyl)-amino-6,7-dimethoxyquinazoline (P-270):

Yield 75.37%; m.p. 225.0-230.0° C. ¹H NMR(DMSO-d₆): δ 11.74(s, 1H, —NH),

8.79(s, 1H, 2-H), 8.33(s, 1H, 5-H), 8.05-7.13(m, 4H, 3′,4,5,6′-H), 7.44(s, 1H, 8H),

4.01(s, 6H, —OCH₃). UV(MeOH): 219.0, 222.0, 253.0, 342.0 nm.

IR(KBr)ν_max: 3165, 3027, 2827, 1639, 1572, 1501, 1434, 1275, 1070 cm⁻¹.

GC/MS m/z 408(M⁺ + 1, 3.47), 407(M⁺, 15.28), 406(M⁺ − 1, 3.47), 281(33.33),

280(M⁺ —I, 100.00), 264(50.00), 207(34.72). Found: C, 43.62; H, 3.60; N, 9.42.

C₁₆H₁₄IN₃O₂.HCl requires: C, 43.34; H, 3.38; N, 9.48%.

4-(3′-Iodophenyl)-amino-6,7-dimethoxyquinazoline (HI-P271):

Yield 79.85%; m.p. 235.0-242.0° C. ¹H NMR(DMSO-d₆): δ 11.43(s, 1H, —NH),

8.88(s, 1H, 2-H), 8.33(s, 1H, 5-H), 8.13(s, 1H, 2′-H), 7.80-7.26(m, 3H, 4′,5′,6′-H),

7.35(s, 1H, 8H), 4.02(s, 3H, —OCH₃), 4.00(s, 3H, —OCH₃).

UV(MeOH): 203.0, 210.0, 228.0, 251.0, 331.0 nm. (KBr)ν_max: 3191, 3022, 2940, 2836,

2576, 1629, 1516, 1444, 1276, 1153, 1060 cm¹. GC/MS m/z 406(M⁺, 1.52),

405(M⁺ − 1, 6.22), 281(35.33), 207(100.00). Found: C, 43.55; H, 3.43; N, 9.32.

C₁₆H₁₄IN₃O₂.HCl requires: C, 43.34; H, 3.38; N, 9.48%.

4-(4′-Hydroxy-3,5-diiodophenyl)-amino-6,7-dimethoxy-quinazoline (HI-P294:

Yield 77.47%; m.p. 259.0-260.0° C. ¹H NMR(DMSO-d₆): δ 11.13(s, 1H, NH),

9.73(s, 1H, —OH), 8.87(s, 1H, 2-H), 8.16(s, 1H, 5-H), 8.09(s, 2H, 2′,6′-H),

7.28(s, 1H, 8H), 3.98(s, 6H, —OCH₃). UV(MeOH)λ_max(ε):. 217.0, 227.0, 252.0 nm.

IR(KBr)ν_max: 3457, 3201, 2934, 2832, 2566, 1629, 1562, 1521, 1439, 1275, 1075 cm⁻¹.

GC/MS m/z: 422(M⁺ —I, 33.53), 405(7.50), 281(86.67), 221(51.80), 207(91.30).

Found: C, 32.60; H, 2.50; N, 6.92. C₁₆H₁₃I₂N₃O₃.HCl requires: C, 32.82; H, 2.39; N, 7.18%.

4-(4′-Hydroxy-3′-iodophenyl)-amino-6,7-dimethoxyquinazoline (HI-P299)

Yield 71.59%; m.p. 248.0-250.0° C. ¹H NMR(DMSO-d₆): δ 11.32(d, 1H, NH),

10.62(s, 1H, —OH, 8.79(s, 1H, 2-H), 8.26(s, 1H, 5-H), 7.98-6.98(m, 3H, 2′,3′,6′-H),

7.32(s, 1H, 8H), 3.98(s, 3H, —OCH₃), 3.97(s, 3H, —OCH₃). UV(MeOH)λ_max(ε): 217.0 ,

227.0, 252.0 nm. IR(KBr)ν_max: 3411, 2975, 2730, 2366, 1634, 1573, 1501, 1429, 1229,

1075 cm⁻¹. GC/MS m/z: 406(M⁺ − 1, 3.33), 405(M⁺ − 2, 7.50),

281(M⁺ − 1 —I, 26.67), 253(11.80), 207(100.00).

Found: C, 41.96; H, 3.40; N, 8.98. C₁₆H₁₄IN₃O₃.HCl requires: C, 41.83; H, 3.26; N, 9.15%.

4-(4′-Iodophenyl)-amino-6,7-dimethoxyquinazoline (HI-P300):

Yield 85.24%; m.p. 240.0-242.0° C. ¹H NMR(DMSO-d₆): δ 11.51(s, 1H, NH),

8.82(s, 1H, 2-H), 8.37(s, 1H, 5-H), 7.81(d, 2H, J=8.4 Hz, 2′,6′-H), 7.55(d, 2H, J=8.4 Hz,

3′,5′-H), 7.35(s, 1H, 8H), 4.01(s, 3H, —OCH₃), 3.98(s, 3H, —OCH₃).

UV(MeOH): 217.0, 227.0, 252.0 nm. IR(KBr)ν_max: 3211, 3032, 2832, 2720, 1629, 1573,

1501, 1434, 1235, 1153, 1070 cm⁻¹. GC/MS m/z 406(M⁺ − 1, 3.33),

405(M⁺ − 2, 7.50), 281(M⁺ − 1 —I, 26.67), 253(11.80), 207(100.00).

Found: C, 43.40; H, 3.39; N, 9.36. C₁₆H₁₄IN₃O₂.HCl. requires: C, 43.34; H, 3.38; N, 9.48%.

Example 5

OH Group Substituted Quinazoline Compounds

OH group substituted quinazoline derivatives were synthesized and characterized as

discussed above for Example 1. The structures and physical data are shown below:

No	Name	Structure	Formula	MW


1	P-93	C₁₆H₁₄ClN₃O₃	331

2	P-97	C₁₆H₁₃Br₂N₃O₃	455

3	P-131	C₁₆H₁₅N₃O₃	297

4	P-132	C₁₆H₁₅N₃O₃	297

5	P-133	C₁₉H₁₆N₄O₃	348

6	P-150	C₁₅H₁₄N₄O₃	298

7	P-154	C₁₆H₁₄BrN₃O₃	376

8	P-180	C₁₆H₁₅N₃O₃	297

9	P-182	C₁₇H₁₅N₃O₅	341

10	P-189	C₁₆H₁₃Cl₂N₃O₃	365

11	P-190	C₁₇H₁₆BrN₃O₃	389

12	P-191	C₁₇H₁₇N₃O₃	311

13	P-192	C₁₆H₁₅N₃O₄	313

14	P-197	C₁₆H₁₄ClN₃O₃	331

15	P-215	C₁₄H₁₃N₅O₄	315

16	P-259	C₁₇H₁₇N₃O₃	311

17	P-265	C₁₈H₁₉N₃O₃	325

18	P-266	C₁₈H₁₉N₃O₃	325

19	P-274	C_2OH₁₇N₃O₃	347

20	P-275	C₂₀H₁₇N₃O₃	347

21	P-276	C₁₈H₁₉N₃O₃	325

22	P-277	C₂₈H₂₃N₃O₃	449

23	P-278	C₁₆H₁₄ClN₃O₃	331

24	P-289	C₁₈H₁₉N₃O₅	357

25	P-292	C₂₀H₁₇N₃O₃	341

26	P-293	C₂₀H₁₇N₃O₃	341

27	P-294	C₁₆H₁₃I₂N₃O₃	549

28	P-229	C₁₆H₁₄IN₃O₃	423

29	P-312	C₁₆H₁₄N₄O₅	342

30	P-313	C₁₆H₁₄N₄O₅	342

31	P-315	C₁₆H₁₄N₄O₅	342

32	P-323	C₁₆H₁₄N₄O₅	342

4-(3′-Chlooro-6′-hydroxylphenyl)amino-6,7-dimethoxyquinazoline (HI-P93)

yield 93.08%; m.p. 295° C.(dec). ⁻H NMR-DMSO-d₆: δ 10.14(s, 1H, —NH), 9.16(s, 1H, —OH),

8.37(s, 1H, 2-h), 7.78(s, 1H, 5H), 7.57(d, 1H, J_2′,2′=2.4 Hz, 2′-H), ), 7.16(s, 1H, 8-H),

7.07(dd, 1H, J_2′,4′=2.4 Hz, J_4′,5′=8.7 Hz, 4′-H), 6.92(d, 1H, J_4′,5′=8.7 Hz, 5′-H),

3.93(s, 3H, —OCH₃), 3.92(s, 3H, —OCH₃. UV(MeOH): 205, 229.0, 251.0, 320.0 nm.

IR(KBr)ν_max: 3500(br), 3430, 2835, 1622, 1512, 1432, 1259 cm⁻¹.

GC/MS m/z 333(M⁻ = 2, 13.41), 332(M⁻ = 1, 9.73), 331(M⁺, 39.47), 314(M⁺ —OH, 100.00),

298(7.64). Found: C, 52.25; H, 4.07; N, 11.39, C₁₆H₁₄ClN₃O₃.HCl requires: C, 52.32; H, 4.09;

N, 11.44%.

4-(3′,5′-Dibromo-4′-hydroxylphenyl)-amino-6,7-dimethoxyquinazoline-(HI-P97).

Yield 72.80%; m.p. > 300.000. ¹H NMR(DMSO-d₆): δ 9.71(s, 1H, —NH), 9.39(s, 1H, —OH),

8.48(s, 1H, 2-h), 8.07(s, 2H, 2′,6′-H), 7.76(s, 1H, 5-H), 7.17(s, 1H, 8-H), 3.94(s, 3H, —OCH₃,

3.91(s, 3H, —OCH₃). UV(MeOH): 208.0, 210.0, 245.0, 320.0 nm;

IR(KBr)ν_max: 3504(br), 3419, 2868, 1627, 1512, 1425, 1250, 1155 cm⁻¹;

GC/MS m/z 456(M¹= 1, 54.40), 455(M⁻, 100.00), 454(M ⁻ 1, 78.01), 439(M⁻ —OH, 7.96),

376(M⁻ + 1 —Br, 9.76), 375(M⁻ Br, 10.91), 360(5.23). Anal. (C₁₆H₁₃Br₂N₃O₃) C, H, N.

4-(4′-Hydroxylphenyl)-amino-6,7-dimethoxyquinazoline (HI-P131):

yield 84.29%; m.p. 245.0-248.0° C. IR(KBr)ν_max: 3428, 2836, 1635, 1516, 1443, 1234 cm:

¹H NMR(DMSO-d₆: δ 11.21(s, 1H, —NH), 9.70(s, 1H, —OH), 8.74(s, 1H, 2-h), 8.22(s, 1H, 5-h),

7.40(d, 2H, J−8.9 Hz, 2′,6′-H), 7.29(s, 1H, 8-H), 6.85(d, 2H, J=8.9 Hz, 3′,5′-H),

3.98(s, 3H, —OCH₃, 3.97(s, 3H, —OCH₂). GC/MS m/z 298(M⁻ = 1, 100.00), 297(M⁻, 26.6),

296(M⁺ − 1, 12.5). Anal. (C₁₆H₁₅N₃O₃HCl) Cl, H, N.

4-(2′-Hydroxylphenyl)-amino-6,7-dimethoxyquinazoline (HI-P132):

yield 82.49%; m.p. 255.0-258.0° C. IR(KBr)ν_max: 3500(br), 3425, 2833, 1625, 1512, 1456, 1251,

1068 cm⁻¹. ¹H NMR(DMSO-d₆): δ 9.78(s, 1H, —NH), 9.29(s, 1H, —OH), 8.33(s, 1H, 2-h),

7.85(s, 1H, 5-H), 7.41-6.83(m, 4H, 3′,4′,5′,6′-H), 7.16(s, 1H, 8-H), 3.93(s, 3H, —OCH₃,

3.92(s, 3H, —OCH₃), 280(M⁺ —OH, 10.0). Anal. (C₁₆H₁₅N₃O₃, HCl) C, H, N.

4-[(8′-Hydroxyquiline)-5′-Jamino-6,7-dimethoxyquinazoline (HI-P133)

yield 83.51%; m.p. 238.0-239.0° C. ₁H NME(DMSO-d₆: δ 10.12(br, s, 1H, —NH),

8.93-7.09 M, 8H, 2,5,2,2′,3′,4′,6′,7′-H), 4.04(s, 3H, —OCH₃), 3.96(s, 3H, —OCH₃).

UV(MeOH): 204.0, 245.0, 332.0 nm. IR(KBr)ν_max: 3425(br), 2935, 1632, 1510, 1437, 1273 cm⁻¹.

GC/MS m/z 349(M⁻ = 1, 100.00), 348(m+, 26.56), 307(38.50), 289(21.00).

4-[(3′-Hydroxylpyridine)-2′]-amino-6,7-dimethoxyquinazoline (HI-P150)

Yield 78.65%; m.p. 185.0-187.0° C. ¹H NMR(DMSO-d₆): δ 10.08(br, s, 1H, —NH),

8.52(s, 1H, 2-H), 7.88-7.86(m, 1H, 6′-H), 7.60(s, 1H, 5-H), 7.39-7.35(m, 1H, 4′-H),

7.32(s, 1H, 8-H), 6.63-6.58(m, 1H, 5′-H), 5.96(s, 1H, —OH), 3.97(s, 3H, —OCH₃),

3.94(s, 3H, —OCH₃). UV(MeOH): 204.0, 238.0, 321.0 nm. IR(KBr)ν_max: 3500, 3446, 2960,

1475, 1236, 1375, 1182 cm⁻¹. GC/MS m/z 299(M⁻ = 1, 100), 298(M⁺, 34),

289(11), 291(9). Found: C, 60.26; H, 4.81; N, 18.68. C₁₅H₁₄N₄O₅,

requires: C, 60.26; H, 4.81; N, 18.68%.

4-(3′-Bromo-4′-hydroxylphenyl)-amino-6,7-dimethoxyquinazoline (HI-P154);

yield 89.90%; m.p. 233.0-233.5° C. ¹H NMR(DMSO-d₆): 10.08(s, 1h, —NH), 9.38(s, 1H, —OH),

8.40(s, 1H 2-H), 7.89(d, 1H, J_2′,6′=2.7 Hz, 2′-H), 7.75(s, 1H, 5-h), 7.55(dd, 1H, J_5′,6′=9.0 Hz,

3.92(s, 3H, —OCH₃), 3.90(s, 3H, —OCH₃). UV(MeOH): 203.0, 222.0, 25.0, 335.0 nm.

IR(KBr)ν_max: 3431(br), 2841, 1624, 1498, 1423, 1244 cm⁻¹. GC/MS m/z 378(M⁺ = 2, 90.68),

377(M⁺ = 1, 37.49), 376(M⁺, 100.00), 360(MK⁺, 3.63), 298(28.86), 282(6.65).

Anal. (C₁₆H₁₄BrN₃O₃, HCl) C, H, N.

4-(3′-Hydroxyphenyl)-amino-6,7-dimethoxyquinazoline (HI-P180)

Yield 71.55%; m.p. 256.0-258.0° C. IR(KBr)ν_max: 3394, 2836, 1626, 1508, 1429, 1251 cm⁻¹.

¹H NMR(DMSO-d₆): 9.41(s, 1H, —NH), 9.36(s, 1H, —OH), 8.46(s, 1H, 2-H), 7.84(s, 1H, 5-H),

7.84-6.50(m, 4H, 2′,4′,5′,6′-H), 7.20(s, 1H, 8-H), 3.96(s, 3H, —OCH³), 3.93(s, 3H —OCH₃).

GC/MS m/z: (C₁₆H₁₅N₃O₃.HCl) C, H, N.

4-(4′-Hydroxyl-3′-Carboxyphenyl)-amino-6,7-dimethoxyquinazoline (HI-P182)

Yield 79.25%; m.p. > 300.0° C. ⁻H NMR(DMSO-d₆)I: δ 10.53(s, 1H, —NH), 8.53(s, 1H, 2-H),

8.10-78.2(m, o3H, 2′, 5′, 6′, —H), 7.26(s, 1H, 5-H), 6.9(s, 1H, 80H), 4.01(s, 3H, —OCH₃),

3.99(s, 3H, —OCH₃). UV(MeOH): 210.0, 239.0, 335.0 nm. IR(KBr)ν_max: 3421, 2839, 1686,

1631, 1508, 1491, 1280 cm⁻¹. GC/MS m/z 341(M⁺, 7.91), 323(M⁺ —OH, 12.19),

297(M⁺ —COOH, 12.35), 296(M⁺ —COOH − 1.1760), 295(M⁺ —COOH − 2, 28.65),

206(11.28).

4-(4′-Hydroxyl-3′-5′-dicholophenyl-6,7-dimethoxyquinazoline (HI-P189)

Yield 79.45%; m.p. 293.0-295.0° C. ¹H NMR(DMSO-d₆): 11.32(s, 1H, —NH),

10.34(a, 1H, —OH), 8.87(s, 1H, 2-H), 8.29(s, 1H, 5-H), 7.90(s, 2H, 2′,6′-H), 7.32(s, 1H, 8-H),

4.01(s, 3H, —OCH₃), 3.99(s, 3H, —OCH₃). UV(MeOH): 213.0, 232.0, 250.0, 335.0 nm.

IR(KBr)ν_max: 3479, 2564, 1641, 1579, 1429, 1282, 1147 cm⁻¹. GC/MS m/z 367(M⁺ + 2; 66.57),

366(M⁺ + 1, 75.91), 365(M⁺, 100.00), 364(M⁺ − 1, 94.08), 349(M⁻ OH, 11.16.

Anal. (C₁₆H₁₃Cl₂N₃O₃) C, H, N. Found: C, 48.93; H, 4.51; N, 10.00. —H₁—Cl₂N₃O₃.HCl

requires: C, 48.80; H, 4.31; N, 10.04%.

4-(4′-Bromo-6′-Hydroxymethylphenyl)-amino-6,7-dimethoxyquinazoline (HI-P190)

Yield 7o3.08%; m.p. 222.0-223.0° C. ¹H NMR(DMSO-d₆): δ 11.30(s, 1H, —OH),

8.22(s, 1H, —NH)O, 7.77.7.23(m, 5H, 5,8,2′,3′,5′-H), 4.49(s, 2H, PhCH₂—H),

4.01(s, 3H, —OCH₃), 3.90(s, 3H, —OCH₃). UV(MeOH): 207.0, 250.0, 332.0 nm.

IR(KBr)ν_max: 3446, 2829, 2752, 1652, 1560, 1471, 1365, 1280 cm⁻¹.

GC/MS m/z 391(M⁻ = 1, 29.33), 389(M⁻, 29.82),

360(M⁻ CH²OH, 50.76), 358(52.39), 311(18.33), 280(43.20), 206(62.80), 191(100.00).

Anal. (C¹⁷H¹⁶BrN₃O_3.HCl) C, H, N.

4-(6′-Hydroxymethylphenyl)-amino-6,7-dimethosyquinazoline (HI-P191)

Yield 78.45%; m.p. 215.0-218.0° C. ¹H NMR(DMSO-d₆): δ 11.54(s, 1H, —NH)O,

8.70(s, 1H, 2-H), 8.34(s, 1H, 5-H), 7.62-7.33(m, 4H, 3′,4′,5′,6′-H), 7.39(s, 1H, 8-H),

4.49(s, 2H, PhCH₂OH), 3.99(s, 3H, —OCH₃), 3.98(s, 3H, —OCH₃).

UV9MeOH): 209.0, 224.0, 246.0, 335.0 nm. IR(KBr)ν_max: 3421, 2941, 1675, 2606, ′128, 1508,

1437o, 1244 cm⁻¹. GC/MS m/z 311(M⁻, 38.07), 310(M⁻ − 1, 27.04),

28o0(M⁻ CH₂OH, 100.00), 206(17.24), 191(51.34).

4-(2′,4′-Dihydroxyphenyl)-amino-6,7-dimethoxyquinazoline (HI-P192)

Yield 86.25%; m.p. 240.0° C.(dec). ¹H NMR(DMSO-d₆): 10.92(s, 1H, —NH), 976(s, 1H, —OH),

9.59(s, 1H, —OH), 8.67(s, 1H, 20H), 81.9(s, 1H, 8-H), 7.36(s, 1H, 50H),

705(d, 1H, J−8.7 Hz, 1′-H), 6.51(s, 1H, 5′-H), 6.31(d, 1H, J−8.7 Hz, 3′-H), 3.98(s, 6H, —OCH₃).

UV(MeOH): 206.0, 209.0, 223.0, 250.0, 342.0, 486 nm. IR(KBR)ν_max: 3391, 3139, 2938, 2850,

1633, 1607, 1567, 1509, 1447, 1359, 1220, 1189, 1055 cm⁻¹. GC/MS m/z: 314(M⁻ = 1, 13.00),

313(m⁻, 72.80), 312(m⁺ − 1, 10.20), 296(524), 206(17.50).

4-(2′,3′-Dihydroxyphenyl)-amino-6,7-dimethoxyquinazoline (HI-P192)

Yield 86.25%; m.p 240.0° C.(dec). ¹H NMR(DMSO-d₆): 10.00(s, 1H, —NH), 9.37(s, 1H, —OH),

8.41(s, 1H, 2-H), 7.78(s, 1H, 5-H), 7.49(d, 1H, J_2′,3′=2.7 Hz, 2′-H),

7.55(dd, 1H, J_5′,6′=9.0 Hz, J_2′,6′=2.7 Hz, 6′-H),

7.16(s, 1H, 8-H), 6.97(d, 1H, J_5′,6′=9.0 Hz, 5′-h), 3.93(s, 3H, —OCH₃),

3.91(s, 3H, —OCH₃). UV9MeOH): 209.0, 224.0, 249.0, 330.0 nm. IR(KBr)ν_max: 3448, 2842,

1623, 1506, 1423, 1241 cm⁻¹. GC/MS m/z: 341(M⁺, 100.00), 326(M⁻ CH₃, 98.50),

310(M⁺ —OCH₃, 12.5), 295(9.0), 189(13.5), 155(13.8). Found: C, 52.35; H, 4.16; Cl, 19.15;

N, 11.39. C₁₆H₁₄ClN₃O₃HCl requires: C, 52.32; H, 4.09; Cl, 19.07; N, 11.44%.

4-(2′,4′-Dihydroxyl-1′,3′-diazine-5′)-amino-6,7-dime-thoxyquinazoline (HI-P215)

(Yield 89.23%, m.p. > 300.0° C.) ¹H NMR(DMSO-d₆): δ 8.59(s, 1H, 2-H), 7.89(s, 1H, 5-H),

7.60(d, 1H, 6′-H), 7.09(s, 1H, 8-H), 3.78(s, 3H, —OCH₃), 3.76(s, 3H, —OCH₃).

UV(MeOH): 222.0, 246.0, 331.0 nm. IR(KBr)ν_max: 3446, 3212, 3057, 1750, 1682, 1620,

1590, 1511, 1420, 1265 cm⁻¹. GC/MS m/z: 315(M⁻ .57.52), 206(46.50), 191(18.21), 127(100.00).

4-(3′-Hydroxymethylphenyl)-amino-6,7-dimethoxyquina-zoline (HI-P259)

Yield 74.28%; m.p. 230.0-232.0° C. ¹H NMR(DMSO-d₆): δ 11.29(s, 1H, —NH),

8.83(s, 1H, 2-H)I, 8.28(s, 1H, 5-H), 7.61-7.25(m, 4H, 2′,4′,5′,6′-H), 7.36(s, 1H, 8H)O,

4.57(s, 2H, —CH2OH), 4.02(s, 3H, —OCH₃), 4.00(s, 3H, —OC₃).

UV(MeOH): 207.0, 224.0, 251.0, 334.0 nm. IR(KBr)ν_max: 3500, 3029, 1639, 1589, 1514,

1456, 1284 cm⁻¹. GC/MS m/z: 281(M⁻ + 1 —CH₂OH, 54.00),

280(M⁻ —CH2OH, 100.00). Found: C, 58.68; H;, 5.30; N, 12.02.

C₁₆H₁₅N₃O₂.HCl requires: C, 58.79; H, 5.19; N, 12.10%.

4-[4′-(2″-Hydroxylethylphenyl)]-amino-6,7-dimethoxyqui-nazoline (HI-P265)

Yield 92.30%; m.p. 235.0-240.0° C. ¹H NMR(DMSO-d₆): δ 11.44(s, 1H, —NH),

8.79(s, 1H, 2-H), 8.34(s, 1H, 5-h)I, 7.56(d, 2H, J=8.1 Hz, 2′,6′-H),

7.34(d, 2H, J−8.1 Hz, 3′,5′-H), 7.31(s, 1H, 8H), 4.00(s, 3H, —OCH₃),

3.99(s, 3H, —OCH₃), 3.64(t, 2H, j=6.9 Hz, 1″-H)I,

2.77(t, 2H, J=6.9 Hz, 2″-H). UV(MeOH): 204.0, 226.0, 251.0, 335.0 m. IR(KBr)ν_max: 3361, 3015,

27o6o7, 1628, 1581, 1514, 1432, 1282 cm⁻¹. GC/MS m/z: 281(17.00), 253(10.00), 207(100.00).

4-[2′-(2″-Hydroxylethylphenyl)]-amino-6,7-dimethoxyqui-nazoline (HI P266)

Yield 87.69%; m.p/ 228.0-230.0° C. ¹H NMR-DMSO-d₆): δ 11.32(s, 1H, —NH),

8.74(s, 1H, 2′-H), 8.13(s, 1H, 5-H), 7.46-7.34(m, 4H, 3′,4′,5,6′-H), 7.32(s, 1H, 8H),

4.00(s, 3H, —OCH₃), 3.99(s, eH, —OCH₃), 3.58(t, 2H, J−7.2 Hz, 1″-H),

2.75(t, 2H, J=7.2 Hz, 2″-H). UV(MeOH): 210.0, 226.0, 249.0, 332.0 nm.

IR(KBr)ν_max: 3366, 3226, 3056, 2917o, 2685, 21638, 1571, 1514, 1467, 1277 cm⁻¹.

GC/MS m/z: 281(20.00), 253(9.00), 207(100.00).

4-(1′-Naphthol-4′)-amino-6,7-edimethoxyquinazoline (HI-P274)

Yield 79.26; m.p. 205.0-208.0° C. ¹H NMR-DMSO-d₆): δ 11.64(s, 1H, —NH),

10.61(s, 1H, —OH), 8.59(s, 1H, 2-h), 8.41(s, 1H, 5-H), 8.22-6.98(m, 5H, 3′,5′,6′,7′,8′-H),

7.40(s, 1H, 8H), 4.00(s, 3H, —OCH₃), 3.99(s, 3H, —OCH₃). UV9MeOH): 208.0, 215.0, 225.0,

240.0, 330.0 nm. IR(KBr)ν_max: 3438, 3211, 3061, 2932, 2834, 1633, 1576, 1509, 1437, 1380,

1276, 1215 cm⁻¹. GC/MS m/z: 281(51.00), 253(22.00), 207(88.00).

Found: C, 62.26; H, 4.87; N, 10.77. C₂₀H₁₇N₃O₃.HCl requires: C, 62.66; H, 4.70; N, 10.96%.

4-(2′-Naphthol-1′)-amino-6,7-dimethoxyquinazoline (HI-P275)

Yield 83.17%; m.p. 218.0-220.0° C. ¹H NMR(DMSO-d₆): δ 11.33(s, 1H, —NH),

10.22(s, 1H, —OH), 8.62(s, 1H, 2-H), 8.40(s, 1H, 5-H), 7.98-7.31(m, 6H, 3′,4′,5′,6′,7″8′-H),

7.41(s, 1H, 8H), 4.02(s, 3H, —OCH₂), 4.00(s, 3H, —OCH₃), . UV(MeOH): 206.0, 210,

219.0, 225.0, 230.0, 340.0 nm. IR(KBr)ν_max: 3391, 3165, 3051, 2938, 2840, 1628, 1576, 1504,

1437, 1281, 1215 cm⁻¹. GC/MS m/z: 348(M ^{− +}1, 7.00), 347(M⁻, 100.00),

346(M⁻ 1.22.00), 331(15.00), 330(12.00), 281(23.00), 253(12.00), 207(49.00).

Found: C, 62.91; H, 4.76; N, 10.75. C₂₀H₁N₃O₃.HCl

requires: C, 62.66; H, 4, 70; N, 10.96%.

4-[3′-(1″-Hydroxyethyl)]-amino-6,7-dimethoxyquinazoline (HI-P276)

Yield 79.21%; m.p. 215.0-218.0° C. ¹H NMR(DMSO-d₆): δ 11.40(s, 1H, —NH),

8.81(s, 1H, 20H), 8.31(s, 1H, 5-H)O, 7.60-7.26(m, 4H, 2′,4′,5′,6′-H), 7.41(s, 1H, 8H),

4.65(q, 1H, J=6.6 Hz, —CH(OH)CH₃), 4.00(s, 3H, —OCH₃), 3.98(s, 3H, —OCH₃),

1.350(d, 3H, J=6.6 Hz, —CH₃). UV9MeOH): 204.0, 216.0, 220.0, 224.0, 250.00, 348.0 nm.

IR(KBr)ν_max: 3407, 3030, 2977, 2840, 1643, 1591 1514, 1463, 1370, 1282, 1230 cm⁻¹.

GC/MS m/z: 325(M⁻ + 1, 67.00), 324(M⁻, 100.00), 323(M⁻ 1.22.00),

308(17.00), 307(56.00), 306(21.00), 281(2.00), 280(8.00), 264(6.00).

4-(4′-Hydroxy-3′,5′-diphenylphenyl)-amino-6,7-dime-hoxyquinazoline (HI-P277)

Yield 76.11%; m.p. 255.0-257.0° C. ¹H NMR_DMSO-d₆): δ 11.50(s, 1H, —NH),

8.80(d, d, 2H, 2′,6′-H), 8.58(s, 1H, 5-H), 7.60-7.30(m, 10H, 3′, 5′, Ph—H), 7.39(s, 1H, 8H),

4.00(s, 3H, —OCH₃), 3.97(s, 3H, —OCH₃), 1.350(d, eH, J=6.6 Hz, —CH₃).

UV(MeOH): 210.0, 214.0, 229.0, 239.0, 345.0, 248.0, 352.0 nm. IR(KBr)ν_max: 3520, 3218,

3023, 2935, 1630, 1562, 1518, 1457, 1281, 1234 cm⁻¹. GC/MS m/z: 281(35.00), 267(6.00),

253(10.00), 207(100.00).

4-(4′-Hydroxyl-2′-chlorophenyl)-amino-6,7-dimethoxy-quinazoline (HI-P2878)

Yield 81.44%; m.p. 245.0-247.0° C. ¹H NMR(DMSO-d₆): δ 11.39(s, 1H, —NH)O,

7.37(s, 1H, 8H), 3.98(s, 3H, —OCH₃), 3.96(s, H₃, —OCH₃).

UV(MeOH): 222.0, 234.0, 239.0, 245.0, 254.0, 348.0 nm.

(R(KBr)ν_max: 3448, 3242, 3144, 3025, 2917, 2834, 1638, 1591, 1514, 1437, 1365,

1277, 1209 cm⁻¹. GC/MS c/z: 332(M⁻ + 1, 5.00), 331(M⁻, 17.00),

330(M⁻ − 1, 5.00), 297(17.00), 296(100.00), 281(18.00), 280o(29.00), 253(9.00).

4-(2′-Hydroxy-naphthyl-3′)-amino-6,7-dimethoxyquinazolin (HI-P292)

Yield 87.41%; m.p. 277.0-279.0° C. ¹H NMR(DMSO-d₆): δ 11.38(s, 1H, —NH)O,

10.35(s, 1H, —OH), 8.73(s, 1H, 2-H), 8.25(s, 1H, 5-H), 7.93-7.30(m, 6H, 1′,4′,5′,6′,7′,8′-H),

7.37(s, 1H, 8H)O, 4.00(s, 6H, —OCH₃). UV(MeOH): 204.0, 221.0, 224.0, 230.0,

256.0, 344.0 nm. IR(KBr)ν_max: 3479, 3386, 3036, 2901, 1632, 1581, 1504, 1437, 1281 cm⁻¹.

GC/MS m/z: 281(41.00), 253(11.00), 207(100.00). Found: C, 62.87; H;, 4.83; N, 1o0.78.

C₂₀H₁N₃O₃.HCl requires: C, 62.66; H, 4.70, N, 10.96%.

4-(1′-Hydroxy-naphthyl-5′)-amino-6,70-dimethoxyquina-zoline (HI-P293)

Yield 87.21%; m.p. 204.0-205.0° C. ¹H NMR(DMSO-d₆): δ 11.73(s, 1H, —NH),

10.43(s, 1H, —OH), 8.65(s, 1H, 2-H, 8.38(s, 1H, 5-H), 8.21-6.95(m, 6H, 2′,3′,4′,6′,7′,8′-H),

7.33(s, 1H, 8H)O, 4.00(s, 6H, —OCH₃). UV9MeOH): 204.0, 214.0, 224.0, 229.0, 235.0 348 nm.

IR(KBrν_max: 3449, 3335, 3102, 2927o, 1633, 1571, 1509, 1437, 1287 cm⁻¹.

Found: C, 62.23; H, 4.96; N, 10.89. C₂₀H₁₇N₃O₃.HCl requires. C, 62.66; H, 4.70; N, 10.96%.

4-(4′-Hydroxy-3,5-diiodophenyl)-amino-6,7-dimethoxy-quinazoline (HI-P294)

Yield 77.47&; m.p. 259.0-260.0° C. ¹H NMR(DMSO-d₆): δ 11.13(s, 1H, NH),

9.73(s, 1H, —OH), 8.87(s, 1H, 2-H), 8.16(s, 1H, 5-H), 8.09(s, 2H, 1′,6′-H), 7.28(s, 1H, 8H),

3.98(s, 6H, —OCH₃),. UV(MeOH)λ_max: 217.0, 227.0, 252.00 nm.

IR(KBrν_max: 3457, 3201, 2934, 2832, 2566, 1629, 1562, 1521, 1439, 1275, 1075 cm⁻¹.

GC/MS m/z: GC/MS m/z 422(M⁻ 1.33.53), 405(7.50), 281(86.67), 221(51.80), 207(91.30).

Found: C, 32.60; H, 2.50; N, 6.92. C₁₆H₁₃I₂N₃O₃.HCl requires: C. 32/82.′ J. 2.39; N, 7.18%.

4-(4′-Hydroxy-3′-iodophenyl)-amino-6,7-dimethoxyquinazoline (HI-P299)

Yield 71.59%; m.p. 248.0-250.0° C. ¹H NMR(DMSO-d₆): δ 11.32(d, 1H, NHO),

7.32(s, 1H, 8H), 3.98(s, 3H, —OCH₃), 3.97(s, 3H, —OCH₃).

UV(MeOH)λ_max(ε): 217.0, 227.0, 252.0 nm.

IR(KBr)ν_max: 3411, 2975, 2730, 2366, 1634, 1573, 1501, 1429, 1229, 1075 cm⁻¹.

GC/MS m/z: 406(M⁻ 1.3.33), 405(M⁻ 2, 7.50), 281(M⁺ − 1 —I, 26.67), 253(11.80), 207(100.00).

TABLE 5


Fluoroquinazoline Derivatives

(HI-P352)		(HI-P353)
No	R	Formular	MW

HI-P144	2-F, 3-F, 5-F, 6-F, 4-Br	C₁₆H₁₀BrF₄N₃O₂	432
HI-P214	2-F, 4-Br	C₁₆H₁₃BrFN₃O₂	378
HI-P218	3-CF₃	C₁₇H₁₄F₃N₃O₂	349
HI-P219	4-OCF₃	C₁₇H₁₄F₃N₃O₃	365
HI-P221	4-F	C₁₆H₁₄FN₃O₂	299
HI-P223	4-CF₃	C₁₇H₁₄F₃N₃O₂	349
HI-P224	3-F	C₁₆H₁₄FN₃O₂	299
HI-P228	2-CF₃	C₁₇H₁₄F₃N₃O₂	349
HI-P232	4-SO₂F	C₁₆H₁₄FN₃O₄S	363
HI-P264	2-F	C₁₆H₁₄FN₃O₂	299
HI-P352	*	C₂₅H₂₀F₆N₄O₂	522
HI-P353	*	C₂₅H₂₀F₆N₄O₂	522
HI-P364	3-OCF₃	C₁₇H₁₄F₃N₃O₃	365
HI-P365	2-OCF₃	C₁₇H₁₄F₃N₃O₃	365
HI-P366	3-CF₃, 5-CF₃,	C₁₈H₁₃F₆N₃O₂	417
HI-P367	2-CF₃, 5-CF₃,	C₁₈H₁₃F₆N₃O₂	417
HI-P369	3-F, 4-OH	C₁₆H₁₄FN₃O₃	315
HI-P408	3-F, 5-F, 4-OH	C₁₆H₁₃F₂N₃O₃	333

HI-P352

HI-P353

Example 6

Fluorine Substituted Quinazoline Compounds

Fluorine substituted quinazoline derivatives were synthesized and characterized as

No	Name	Structure	Formula	MW


1	P-144	C₁₆H₁₀BrF₄N₃O₂	432

2	P-214	C₁₆H₁₃BrFN₃O₂	378

3	P-218	C₁₇H₁₃F₄N₃O₂	367

4	P-219	C₁₇H₁₄F₃N₃O₃	365

5	P-221	C₁₆H₁₄FN₃O₂	299

6	P-223	C₁₇H₁₄F₃N₃O₂	349

7	P-224	C₁₆H₁₄FN₃O₂	299

8	P-228	C₁₇H₁₄F₃N₃O₂	349

9	P-232	C₁₆H₁₄F₂SN₃O₄	363

10	P-264	C₁₆H₁₄FN₃O₂	299

11	P-352	C₂₅H₂₀F₆N₄O₂	522

12	P-353	C₂₅H₂₀F₆N₄O₂	522

13	P-364	C₁₇H₁₄F₃N₃O₃	365

14	P-365	C₁₇H₁₄F₃N₃O₃	365

15	P-366	C₁₈H₁₃F₆N₃O₂	417

16	P-367	C₁₈H₁₃F₆N₃O₂	417

17	P-369	C₁₆H₁₄FN₃O₃	315

18	P-408	C₁₆H₁₃F₂N₃O₃	333

4-(2′,3′,5′,6′-Terrafluoro-4′-bromophenyl)-amino-6,7-dime-thoxyquinazoline (HI-P144)

The yield 78.24%: m.p. 180.0-182.0° C. ¹H NMR(DMSO-d₆): δ 7.78(s, 1H, 2-H),

7.53(s, 1H, 5-H), 6.79(s, 1H, 8-H), 3.81(s, 3H, —OCH₃), 3.3.79(s, 3H, —OCH₃).

Found: C, 41.12; H, 2.41; N, 9.89, C₁₀H₁₀BrF_N₃O₂.HCl.

requires: C, 41.11; H, 2.36; N, 9.93%.

4-(2′-Fluoro-4′-bromophenyl)-amino-6,7-dimethoxyquina-zoline (HI-P214)

The yield 77.21%; m.p. 247.0-252.0 0° C. ¹H NMR(DMSO-d₆): δ 8.57(s, 1H, 2-H),

7.91(s, 1H, 5-H), 7.57(d, 1H, 3′-H), 7.34(m, 2H, 5′,6′-H), 7.07(s, 1H, 8-H), 3.78(s, 3H, —OCH₃),

3.77(s, 3H, —OCH₃). UV(MeOH): 204.0, 215.0, 250.0, 330.0 nm.

IR(KBr)ν_max: 3431, 2629, 1633, 1580, 1511, 1420, 1278 cm⁻¹.

GC/MS m/z 379(M⁺ + 1, 34.39), 378(M⁻, 31.33), 377(M⁻ − 1, 39.08),

360(62.05), 359(31.58), 358(62.57), 357(19.81), 299(19.31), 298(100.00), 282(17.88), 240(28.76).

4-(3′-Trifluoromethylphenyl)-amino-6,7-dimethoxyquinazo-line (HI-P218)

The yield 85.61%: m.p. 242.0-245.0 0° C. ¹H NMR(DMSO-d₆): δ 11.09(s, 1H, —NH),

8.67(s, 1H, 2-H), 8.03(s, 1H, 5-H), 7.92-7.43(m, 4H, 2′4′5′,6′-H), 7.10(s, 1H, 8-H),

3.81(s, 3H, —OCH₃), 3.79(s, 3H, —OCH₃). UV(MeOH): 206.0, 276.0, 349.0 nm.

IRν_max(KBr): 3372, 3257, 2935, 1626, 1512, 1380, 1225 cm⁻¹. GC/MS m/z 350(M⁺ + 1, 10.5),

249(M⁻, 85.5), 173(M⁻ − 1, 100.0), 332(10.5), 290(8.8).

4-(4′-Trifluoromethoxylphenyl)-amino-6,7-dimethoxyqui-nazoline (HI-P219)

The yield 83.14%; m.p. 228.0-230.0 0° C. ¹H NMR(DMSO-d₆): δ 11.39(s, 1H, —HN),

8.63(s, 1H, 2-H), 8.18(s, 1H, 5-H), 7.63(t, 2H, 3′,5′-H), 7.27(t, 2H, 2′,6′-H), 7.15(s, 1H, 8-H),

3.81(s, 3H, —OCH₃), 3.78(s, 3H, —OCH₃). UV(MeOH): 209.0, 216.0, 251.0, 332.0 nm.

IR(KBr)ν_max: 3207, 2839, 2762, 1633, 1508, 1480, 1276 cm⁻¹.

GC/MS m/z 366(M⁺ + 1, 12.50), 365(M⁻, 75.00), 364(M⁻ − 1, 100.00),

348(17.50), 319(19), 306(8.00), 207(15.00).

4-(4′-Fluorophenyl)-amino-6,7-dimethoxyquinazoline (HI-P221)

The yield 84.25%: ¹H NMR(DMSO-d₆): δ 11.19(s, 1H, —HN), 8.60(s, 1H, 2-H), 8.08(s, 1H, 5-H),

7.50(t, 2H, 3′-H), 7.13(s, 1H, 8-H), 7.12(t, 2H, 2′,6′-H), 3.79(s, 3H, —OCH₃), 3.78(s, 3H, —OCH₃).

UV(MeOH): 225.0, 251.0, 333.0 nm. IR(KBr)ν_max: 3205, 3007, 2837, 1633, 1580,

1508, 1470, 1220 cm⁻¹. GC/MS m/z 300(M⁺ + 1, 10.76), 299(m⁻, 76.92), 39.8(M⁻ − 1, 100.00),

282(20.00), 253(13.08), 207(3.80). Found: C, 57.17; H, 4.37; N, 12.47, C₁₆H₁₄FN₃O₂.HCl

requires C, 57, 31; H, 4.48; N, 12.54%.

4-(4′-Trifluoromethylphenyl)-amino-6,7-dimethoxyquinazoline (HI-P223)

The yield 91.70%: m.p. 243.0-245.0 0° C. ¹H NMR(DMSO-d₆): δ 11.47(s, 1H, —NH), 8.67(s, 1H, 2-H),

8.23(s, 1H, 5-H), 7.79(d, 2H, J=8.4 Hz, 3′5′-H), 7.61(d, 2H, J=8.4 Hz, 2′6′-H),

7.17(s, 1H, 8-H), 3.82(s, 3H, —OCH₃), 3.78(s, 3H, —OCH₃). GC/MS m/z 350(M⁻ + 1, 11.00),

349(M⁻, 65.00), 348(M⁻ − 1, 100.00), 332(18.50), 303(10.00), 207(18.50). Found: C, 53.01;

H, 3.94; N, 10.88. C₁—H₁₄F₃N₃O₂HCl requires C, 52.98; H, 3.90; N, 10.91%.

4-(4′-Flurophenyl)-amino-6,7-dimethoxyquinazoline (HI-P224)

The yield 88.69%; m.p. 254.0-255.0 0° C. ¹H NMr(DMSO-d₆): δ 11.16(s, 1H, —HN), 8.67(s, 1H, 2-H),

8.09(s, 1H, 5-H), 7.13(s, 1H, 8-H), 7.51-6/94(m, 4H, 2′,3′,5′,6′-H)O, 3.80(s, 3H, —OCH₃),

3.79(s, 3H, —OCH₃). UV(MeOH): 206.0, 226.0, 251.0, 333.0, 343 nm. IR(KBr)ν_max: 3437, 3211, 2619,

1637, 1580, 1500, 1448, 1281 cm⁻¹. GC/MS m/z (300(M⁺ + 1, 8.00), 299(M⁻, 68.00),

398(M ⁻ 1, 100.00), 282(21.60), 253(25.00), 207(80.00),. Found: C, 57.25; H, 4.58; N, 12.42.

C₁₆H₁₄FN₃O₂.Hcl requires C, 57.31; H, 4.48; N, 12.54%.

4-(2′-Trifluoromethylphenyl)-amino-6,7-dimethoxyquinazoline (HI-P228).

The yield 83.57%; m.p. 242.0-245.0 0° C. ¹H NMR(DMSO-d₆): δ 11.58(s, 1H, —HN), o8.76(s, 1H, 2-H),

8.25(s, 1H, 5-H), 7.95-7.62(m, 4H, 3′,4′,5′,6′-H), 7.38(s, 1H, 8H), 4.01(S, 3H, —OCH₃),

3.00(s, 3H, —OCH₃). GC/MS m/z 350(M⁻ + 1, 8.50), 349(M⁻, 32.00), 348(M⁺ − 1.31.50),

328(18.50), 207(5.0)I, 280(M⁺ —CF₃, 100.00), 264(18.50), 207(32.50).

Found: C, 52.71; H, 4.26; N, 10.91%.

4-[4′-benzenesulfanilyl fluoride]-amino-6,7-dimethoxyquinazoline (HI-P232)

Yield 84.02%; m.p. 228.0-230.0° C. ¹H NMR9DMSO-d₆): δ 11.62(s, 1H, —HN), 8.78(s, 1H, 2-H),

8.29(s, 1H, 5-H), 8.12-8.02(m, 4H, 2″,3″,5″,6″-H), 7.21(s, 1H, 8-H), 3.86(s, 3H, —OCH₃),

3.81(s, 3H, —OCH₃). UV(MeOH): 208.0, 215.0, 253.0, 278.0, 338.0 nm.

IR(KBr)ν_max: 3440, 3277, 2571, 1635, 1580, 1516, 1435, 1209 cm⁻¹.

GC/MS m/z: 281(43.00), 253(12.00), 207(100.00). Found: C, 48.13; H, 3.73; N, 10.53.

C₁₆H₁₄FN₃O₄S.HCl requires: C, 48.12; H, 3.76; N, 10.53%.

4-(2′-Fluorophenyl)-amino-6,7-dimethoxyquinazoline (HI-P264)

Yield 73.58%; m.p. 233.0-235.0 0° C. ¹H NMR(DMSO-d₆): δ 11.69(d, 1H, —NH), 8.82(s, 1H, 2-H),

8.37(s, 1H, k 50H), 7.59-7.32(m, 4H 3′,4′5′,6′-H), 7.41(s, 1H, 8H)O, 4.02(s, 3H, —OCH₃),

4.01(s, 3H, —OCH₃). UV(MeOH): 204.0, 226.0, 248.0, 330.0 nm. IR(KBrν_max: 3454, 3032, 2638,

1630, 1589, 1514, 1430, 1291 cm⁻¹. GC/MS m/z 300(M⁺ = 1, 7.00), 299(M⁻, 38.00),

298(M⁻ 1.22.00), 280(M⁻ F, 100.00), 264(15.00), 207(35.00). Found: C, 57.12; H, 4.57; N, 12.45.

C₁₆H₁₄FN₃O₂.HCl requires: C, 57.31; H, 4.48; N, 12.54%.

4-{4′-[2″-(4′′′-Aminophenyl)-hexafluoropropyl]phenyl}-amino-6,7-dimethoxyquinazoline (HI-P352)

Yield, 80.41%, m.p. 280.0-282.0° C. ¹H NMR(DMSO-d₆): δ 11.87(s, 1H, —NH), 8.91(s, 1H, 2-H)I,

8.55-7.18(m, 10H, 5,8,2′,3′,5′,6′,2′′′,3′′′,5′′′,6′′′-H),

4.05(s, 3H, —OCH₃), 4.00(s, 3H, —OCH₃). ¹⁹F NMR(DMSO-d₆): 128.76.

Found: C, 50.33; H, 3.87; N, 9.57. C₂₅H₂₀F₆N₄O₂.2HCl requires: C, 50.50; H, 3.70; N, 9.42%

4-{3′-[2″-(3′′′-Aminophenyl)-hexafluoropropyl]phenyl}-amino-6,7-

dimethoxyquinazoline (HI-P353)

Yield, 83.11%, m.p. 292.0-284.0° C. ¹H NMR(DMSO-d₆): δ 11.68(s, 1H, —NH), 8.81(s, 1H, 2-H),

8.44-7.09(m, 10H, 5,8,2′,4′,5′,6′,2′′′,4′′′,5′′′6′′′-H).

4.00(s, 3H, —OCH₃), 3.97(s, 3H, —OCH₃). ¹⁹F NMR(DMSO-d₆): 129.21.

Found: C, 53.96; H, 3.93; N, 9.77. C₂₅H₂₀F₆N₄O₂.HCl requires: C, 53.76; H, 3.76; N, 10.03%

4-(3′-Trifluoromethoxylphenyl)-amino-6,7-dimethoxyquinazoline (HI-P364)

Yield, 83.25%. m.p. 233.0-235.0° C. ¹H NMR(DMSO-d₆): δ 11.65(s, 1H, —NH), 8.88(s, 1H, 2-H),

8.41(s, 1H, 5-H), 7.86-7.29(m, 4H, 2′,4′,5′,6′-H), 7.36(s, 1H, 8-H), 4.02(s, 3H, —OCH₃).

3.98(s, 3H, —OCH₃). ¹⁹F NMR(DMSO-d₆): 135.37. GC/MS m/z: 366(M⁺ + 1, 11.0),

365(M⁺, 67.0), 364(M⁺ − 1, 100.0). Found: C, 50.93; H, 3.75; N, 10.61.

C₁₇H₁₄F₃N₃O₃.HCl requires: C, 50.97; H, 3.74; N, 10.47%.

4-(2′-Trifluoromethoxylphenyl)-amino-6,7-dimethoxyquinazoline (HI-P365)

Yield, 77.85%. m.p. 235.0-237.0° C. ¹H NMR(DMSO-d₆): δ 11.68(s, 1H, —NH), 8.80(s, 1H, 2-H),

8.32(s, 1H, 5-H), 7.64-7.53(m, 4H, 3′,4′,5′,6′-H), 7.40(s, 1H, 8-H), 3.99(s, 6H, —OCH₃).

¹⁹F NMR(DMSO-d₆): 135.71. GC/MS m/z: 366(M⁻ + 1, 2.0), 365(M⁺, 15.0),

364(M⁺ − 1, 4.0), 281(21.0), 280(M⁻ —OCF₃100). Found: C, 50.83; H, 3.79; N, 10.52.

C_1—H₁₄F₃N₃O₃.HCl requires: C, 50.87; H, 3.74; N, 10.47%.

4-(3′,5′-Ditrifluoromethylphenyl)-amno-6,7-dimethoxyquinazoline (HI-P366)

Yield, 82.88% m.p. 270.0-272.0° C. ¹H NMR(DMSO-d₆): δ 11.87(s, 1H, —NH), 8.97(s, 1H, 2-H),

8.60)s, 2H, 2′,6′-H), 8.43(s, 1H, 5-H), 7.98(s, 1H, 4′-H), 7.35(s, 1H, 8-H), 4.03(s, 3H, —OCH₃).

3.99(s, 3H, —OCH₃). ¹⁹F NMR(DMSO-d₆): XX GC/MS m/z: 418(M⁻ + 1, 19.0),

417(M⁻, 100.0), 416(M⁻ − 1, 73.0), 398(M⁻ —F, 16.0), 398(M⁻ —F, 16.0), 348(M⁻ —CF₃, 16.0).

Found: C, 47.78; H, 3.20; N, 9.26. C₁₈H₁₃F₆N₃O₂.HCl requires: C, 47.68; H, 3.09; N, 9.27%.

4-(4′-Hydroxyl-3′-fluorophenyl)-amino-6,7-dimethoxyquinazoline (HI-P369)

Yield, 84.28%. m.p. 268.0-270.0° C. ¹H NMR(DMSO-d₆: δ 11.36(s, 1H, —NH), 10.13(s, 1H, —OH),

8.80(s, 1H, 2-H), 8.30(s, 1H, 5-H), 7.60-7.02(m, 3H, 2′,5′,6′-H), 7.31(s, 1H, 8-H), 3.99(s, 3H, —OCH₃),

3.97(s, 3H, —OCH₃). ¹⁹F NMR(DMSO-d₆): δ 57.38. Found: C, 54.90; H, 4.28; N, 11.91.

C₁₆H₁₄FN₃O₃.HCl requires C, 54.70; H, 4.27; N, 11.97%.

4-(4′-Hydroxyl-3′,5′-difluorophenyl)-amino-6,7-dimethoxy-quinazoline (HI-P408)

Yield, 83.15%, m.p. 228.0-230.0° C. ¹H NMR(DMSO-d₆): δ 11.46(s, 1H, —NH),

10.39(s, 1H, 2-H), 8.36(s, 1H, 5-H), 7.56, 7.54 (s, s, 2H, 2′,6′-H), 7.33(s, 1H, 8-H), 4.00)s, 3H, —OCH₃),

3.98(s, 3H, —OCH₃). ¹⁹F NMR(DMSO-d₆: δ 60.25, 60.22.

Found: C, 52.04; H, 4.17; N, 11.10. C₁₆H₁₃F₂N₃O₃.HCl. requires C, 52.03; H, 3.79; N, 11.38%.

Example 7

Anti-Tumor Activities of Specific Quinazoline Compounds

The cytotoxicity of the substituted quinazoline derivative compounds against a variety of human tumor cells was evaluated. The relative importance of particular substituent groups on the compounds was also studied. The substituted quinazoline derivative compounds, prepared as described above, were tested, along with DMSO as a control. [0106]
Cytotoxicity Assay [0107]
The cytotoxicity assay of various compounds against human tumor cell lines was performed using the MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide) assay (Boehringer Mannheim Corp., Indianapolis, Ind.). Briefly, exponentially growing tumor cells were seeded into a 96-well plate at a density of 2.5×10[0108] ⁴cells/well and incubated for 36 hours at 37° C. prior to drug exposure. On the day of treatment, culture medium was carefully aspirated from the wells and replaced with fresh medium containing the quinazoline compounds at concentrations ranging from 0.1 to 250 μM. Triplicate wells were used for each treatment.
Human cell lines were obtained from American Type Culture Collection (Rockville, Md.) and maintained as a continuous cell line in Dulbecco's modified Eagles' medium supplemented with 10% fetal bovine serum and antibiotics. Cells used in this study include human leukemia cells (NALM-6 and MOLT-3), human breast cancer cells (BR20), human prostate cancer cells (PC3), and human brain tumor cells (U373). [0109]
The cells were incubated with the various compounds for 24-36 hours at 37° C. in a humidified 5% CO[0110] ₂atmosphere. To each well, 10 μl of MIT (0.5 mg/ml final concentration) was added and the plates were incubated at 37° C. for 4 hours to allow MTT to form formazan crystals by reacting with metabolically active cells. The formazan crystals were solubilized overnight at 37° C. in a solution containing 10% SDS in 0.01 M HCl. The absorbance of each well was measured in a microplate reader (Labsystems) at 540 nm and a reference wavelength of 690 nm. To translate the OD 540 values into the number of live cells in each well, the OD 540 values were compared to those on standard OD 540-versus-cell number curves generated for each cell line. The percent survival was calculated using the formula: $% Survival = \frac{live cell number [test]}{live cell number [control]} \times 100$
The IC[0111] ₅₀values were calculated by non-linear regression analysis.
Detection of Apoptosis [0112]
The demonstration of apoptosis was performed by the in situ nick-end-labeling method using ApopTag in situ detection kit (Oncor, Gaithersburg, Md.) according to the manufacturer's recommendations. Exponentially growing cells were seeded in 6-well tissue culture plates at a density of 50×10[0113] ⁴cells/well and cultured for 36 hours at 37° C. in a humidified 5% CO₂atmosphere. The supernatant culture medium was carefully aspirated and replaced with fresh medium containing unconjugated EGF or EGF-P154 at a concentration of 10, 25, or 50 μg/ml. After a 36 hour incubation at 37° C. in a humidified 5% CO₂incubator, the supernatants were carefully aspirated and the cells were treated for 1-2 minutes with 0.1% trypsi The detached cells were collected into a 15 mil centrifuge tube, washed with medium and pelleted by centrifugation at 1000 rpm for 5 minutes. Cells were resuspended in 50 μl of PBS, transferred to poly-L-lysine coated coverslips and allowed to attach for 15 minutes. The cells were washed once with PBS and incubated with equilibration buffer for 10 minutes at room temperature.
After removal of the equilibration buffer, cells were incubated for 1 hour at 37° C. with the reaction mixture containing terminal deoxynucleotidyl transferase (TdT) and digoxigenin-11-UTP for labeling of exposed 3′-hydroxyl ends of fragmented nuclear DNA. The cells were washed with PBS and incubated with anti-digoxigenin antibody conjugated to FITC for 1 hour at room temperature to detect the incorporated dUTP. After washing the cells with PBS, the coverslips were mounted onto slides with Vectashield containing propidium iodide (Vector Labs, Burlingame, Calif.) and viewed with a confocal laser scanning microscope. Non-apoptotic cells do not incorporate significant amounts of dUITP due to lack of exposed 3-hydroxyl ends, and consequently have much less fluorescence than apoptotic cells which have an abundance of exposed 3′-hydroxyl ends. In control reactions, the TdT enzyme was omitted from the reaction mixture. [0114]
Results: [0115]

The cytotoxicity results for each tested group of compounds is shown in Tables 1-5 below:

TABLE 1


Cytotoxic Activity of Bromo Substituted Quinazoline Compounds
against Leukemic (NALM-6 & MOLT-3) and Breast Cancer (BT-20)

	NALM-6	MOLT-3	BT20
	IC50	IC50	IC50
Drug	(μM)	(μM)	(μM)

HI-P79	142.1	194.9	201.5
HI-P88	>250	>250	>250
HI-P97	>250	>250	26.1
HI-P111	200.6	>250	>250
HI-P154	12.5	9.1	>250
HI-P160	135.2	240.7	25.5
HI-P164	>250	>250	39.2
HI-P190	>250	>250	>250
HI-P210	>250	>250	>250
HI-P211	>250	>250	>250
HI-P212	52.7	54.5	>250
HI-P214	>250	>250	>250
Hl-P222	34.0	48.3	>250
HI-P234	>250	>250	>250
HI-P241	>250	>250	>250
HI-P258	>250	>250	>250
HI-P260	32.4	51.3	82.1
HI-P261	72.6	148.5	218.6
HI-P262	>250	>250	>250

TABLE 2


Cytotoxic Activity of Chloro Substituted Quinazoline Compounds
against Leukemic (NALM-6 & MOLT-3) and Breast Cancer (BT-20)

	NALM-6	MOLT-3	BT20
	IC50	IC50	IC50
Drug	(μM)	(μM)	(μM)

HI-P87	95.9	>104.6	>250
HI-P93	>250	>250	>250
HI-P189	>250	>250	>250
HI-P197	39.3	68.0	136.9
HI-P239	29.6	28.7	25.7
HI-P246	>250	>250	>250
HI-P268	215.2	227.4	121.5
HI-P269	>250	>250	>250
HI-P415	67.9	>250	38.1

[0118]

TABLE 3

Cytotoxic Activity of Iodide Substituted Quinazoline

Compounds against Leukemic (NALM-6 & MOLT-3),

Breast Cancer (BT-20) and Brain Tumor (U373) cells

NALM-6 MOLT-3 BT20 U373

IC50 IC50 IC50 IC50

Drug (μM) (μM) (μM) (μM)

HI-P270 >250 78.9 >250 >250

HI-P271 6.1 9.6 >250 >250

HI-P294 >250 >250 >250 >250

HI-P299 15.4 60.1 >250 >250

HI-P300 58.0 59.1 72.6 116.2

TABLE 4


Cytotoxic Activity of OH Substituted Quinazokline
Compounds against Leukemic (NALM-6 & MOLT-3),
Breast Cancer (BT-20) and Brain Tumor (U373) cells

	NALM-6	MOLT-3	BT20	U373
	IC50	IC50	IC50	IC50
Drug	(μM)	(μM)	(μM)	(μM)

HI-P93	>250	>250	>250	>250
HI-P97	>250	>250	26.1	161.2
HI-P131	32.1	38.6	>250	>250
HI-P154	12.5	9.1	>250	167.4
HI-P189	>250	>250	>250	>250
HI-P190	>250	>250	>250	>250
HI-P192	>250	>250	>250	>250
HI-P197	68.5	63.8	71.5	>250
HI-P294	>250	>250	>250	>250
HI-P299	66.3	51.2	>250	>250

TABLE 5


Cytotoxic activity of fluoro-substituted dimethoxy quinazolines on cancer cells.

	NALM-6	MOLT-3	U373	BT20	PC3
	IC50	IC50	IC50	IC50	IC50
Compound	(μM)	(μM)	(μM)	(μM)	(μM)

HI-P144	28.1 ± 2.6	24.9 ± 3.7	49.5 ± 11.3	63.4 ± 5.5	>250
Hl-P214	>250	>250	>250	>250	>250
HI-P218	37.0 ± 5.8	33.2 ± 3.3	29.9 ± 7.3	37.62 ± 5.2	126.1 ± 5.8
HI-P219	22.3 ± 3.0	41.3 ± 4.4	83.6 ± 6.5	44.2 ± 10.9	58.3 ± 3.2
HI-P221	100.5 ± 4.8	98.73 ± 3.8	28.8 ± 12.7	30.67 ± 7.9	>250
HI-P223	39.5 ± 8.0	40.8 ± 15.1	32.1 ± 3.9	27.56 ± 8.6	>250
HI-P224	20.15 ± 8.1	23.3 ± 7.7	22.4 ± 5.9	58.33 ± 5.8	>250
HI-P228	57.3 ± 24.8	237.1 ± 4.8	>250	>250	>250
HI-P232	41.4 ± 6.9	43.6 ± 2.3	207.7 ± 18.1	70.54 ± 8.2	88.9 ± 17.2
HI-P264	47.0 ± 19.5	70.9 ± 17.3	53.3 ± 6.7	33.33 ± 7.5	>250
HI-P352	7.1 ± 1.8	21.8 ± 1.7	65.5 ± 11.2	50.3 ± 14.8	72.6 ± 2.5
HI-P353	6.1 ± 1.4	17.4 ± 1.5	14.5 ± 7.6	14.1 ± 3.3	64.9 ± 11.9
HI-P364	7.9 ± 1.9	25.3 ± 9.1	27.7 ± 1.2	40.1 ± 8.6	>250
HI-P365	86.5 ± 3.4	110.7 ± 7.5	>250	>250	>250
HI-P366	52.8 ± 14.0	137.2 ± 10.3	55.5 ± 13.2	61.7 ± 12.1	>250
HI-P369	>250	>250	>250	>250	>250
HI-P408	116.3 ± 17.8	228.5 ± 20.8	>250	>250	>250

The compounds were tested for activity against various cancer cells. For example, NALM-6 cells were incubated with HI-P144, HI-P214, HI-P221, HI-P224,.HI-P258, HI—P264, HI-P218, HI-P223, HI-P228, HI-P366, HI-P367, HI-P219, HI-P352, HI-P353, HI-P364 or HI-P365 for 24 hours in 96-well plates and cell survival was determined by MTT assay. The data points represent the means (±SE) values from 3 independent experiments. [0121]
BT-20 breast cancer cells were incubated with HI-P144, HI-214, HI-P221, HI-P224, HI-P258, HI-P264, HI-P218, HI-P223, HI-P228, HI-P366, HI-P367, HI-P219, HI-P352, HI-P353, HI-P364, or HI-P365 for 24 hours in 96-well plates and cell survival was determined by MTT assay. The data points represent the mean (±SE) values from 3 independent experiments. [0122]
Apoptosis was induced by the compounds. The cells were incubated with HI-P353 or HI-P219 for 24 hours, fixed, permeabilized, and visualized for DNA degradation in a TUNEL assay using digoxigenin-UTP-labeling kit. In the color photographs, red fluorescence represents nuclei stained with propidium iodide. Green or yellow (i.e., superimposed red plus green) represents the apoptotic nuclei. Shown are control NALM-6 (FIG. 3A) and BT-20 (FIG. 3B) cells; HI-P353 treated NALM (FIG. 3D) and BT-20 cells (FIG. 3E); and control (FIG. 3C) and HI-P219 treated PC3 cells (FIG. 3F). [0123]

Example 8

Pharmacokinetic Studies

In pharmacokinetic studies, mice were injected either intravenously (i.v.) via the tail vein or intraperitoneally (i.p.) with a bolus dose of 300 μg/mouse (˜12.5 mg/kg=34 μmols/kg) of HI-P131. Blood samples were obtained from the ocular venous plexus by retroorbital venupuncture prior to and at 3 minutes, 5 minutes, 10 minutes, 15 minutes, 30 minutes, 45 minutes and 1 hour, 2 hours, 4 hours, and 8 hours after administration of HI-P131. All collected blood samples were heparinized and centrifuged at 7,000×g for 10 minutes in a microcentrifuge to obtain plasma. The plasma samples were stored at −20° C. until analysis. Aliquots of plasma were used for extraction and HPLC analysis. Pharmacokinetic modeling and pharmacokinetic parameter calculations were carried out using the pharmacokinetics software, WinNonline Program, Version 1.1. (Scientific Consulting Inc., Cary, N.C.). Concentration data were weighted by I/concentration. An appropriate pharmacokinetic model was chosen on the basis of lowest weighted squared residuals, lowest Schwartz criterion (SC), lowest Akaike's Information Criterion (AIC) value, lowest standard errors of the fitted parameters, and dispersion of the residuals. The elimination half-life was estimated by linear regression analysis of the terminal phase of the plasma concentration profile. The area under the concentration time curve (AUC) was calculated by the trapezoidal rule between first (0 h) and last sampling time plus C/k, where C is the concentration of last sampling and k is the elimination rate constant. Systemic clearance (CL) was determined by dividing the dose by the AUC. The apparent volume of distribution at steady-state was calculated using the following equation, V[0124] _ss=Dose·AUMC/(AUC)². Bioavailability (F) was estimated using the equation F(%)=AUC_ip×Dose_iv/AUC_iv×Dose_ip.
HPLC Analysis of Plasma HI-P131 Levels: [0125]
A highly sensitive quantitative HPLC detection method (Chen et al., 1998, [0126] J. Chromatography B (Biomedical sciences), in press) was used to determine the pharmocokinetics of HI-P131. In brief, the HPLC system consisted of a Hewlett Packard (HP) series 1100 equipped with an automated electronic degasser, a quaternary pump, an autosampler, an automatic thermostatic column compartment, diode array detector and a computer with a Chemstation software program for data analysis. A 250×4 mm Lichrospher 100, RP-18 (5 μm) analytical column and a 4×4 mm Lichrospher 100, RP-18 (5 μm) guard column were obtained from Hewlett Packard Inc. (San Fernando, Calif.). Acetonitrile/water containing 0.1% of trifluoroacetic acid (TFA) and 0.1% triethylamine (TEA) (28:72, v/v) was used as the mobile phase. The wavelength of detection was set at 340 nm. Peak width, response time and slit were set at >0.03 minutes, 0.5 seconds and 8 nm, respectively.
For determination of HI-P131 levels, 10 μL of internal standard HI-P154 (50 μM) was added to a 100 μL plasma sample. For extraction, 7 ml chloroform was then added to the plasma sample, and the mixture was vortexed thoroughly for 3 minutes. Following centrifugation (300×g, 5 minutes), the aqueous layer was frozen using acetone/dry ice and the organic phase was transferred into a clean test tube. The chloroform extracts were dried under a slow steady stream of nitrogen. The residue was reconstituted in 100 μL of methanol: water (9:1) and 50 μL aliquot of this solution was used for HPLC analysis. Under the described chromatographic separation conditions, the retention times for HI-P131 and HI-P154 were 5.1 minutes and 9.5 minutes, respectively. At the retention time, HI-P131 and its internal standard HI-P154 were eluted without any interference peaks from the blank plasma. [0127]
HI-P131 was not toxic to mice at intraperitoneal single bolus doses ranging from 0.5 mg/kg to 250 mg/kg. None of the 50 mice treated with HI-P1131 experienced side effects or died of toxicity during the 30 day observation period. In particular, we observed no hematologic side effects such as neutropenia, lymphopenia, or anemia at the tested dose levels. No histopathologic lesions were found in the organs of HI-P131 treated mice that were selectively killed at 30 days and there was no bone marrow hypoplasia or lymphoid cell depletion in spleen and lymph nodes. Thus, the maximum tolerated dose (MTD) of HI-P131 was not reached at 250 mg/kg. We next examined the pharmacokinetic features of HI-P131 in mice. A two-compartment pharmacokinetic model was fit to the pharmacokinetics data obtained following the intravenous (i.v.) or intraperitoneal (i.p.) administration of a single non-toxic 12.5 mg/kg bolus dose of HI-P131. The estimated maximum plasma concentrations (Cma.) of HI-P131 were 85.6 μM after i.v. administration. [0128]

Example 9

Antitumor Activity of Quinazolines In Vivo

To test the anti-tumor activity of quinazolines in vivo, cancer cells were implanted and grown in mice in the presence of quinazoline. [0129]
Inhibition of Breast Cancer Cells [0130]
The left hind legs of CB.17 SCID mice were inoculated subcutaneously with 0.75×10[0131] ⁶MDA-MB-231 human breast cancer cells in 0.1 ml PBS. Twenty-four hours after inoculation, the mice were treated with HI-P353 (10 mg/kg/day x 5 days/week, N=7), or HI-P364 (10 mg/kg/day×5 days/week, N=8), or vehicle (50% DMSO in PBS, N=7) for four weeks. The mice were monitored daily for health status and tumor growth. Measurements were taken on the tumors 3 times a week using a Vernier caliper. Tumor volumes were calculated using the following formula: (width)²×(length/2). Comparisons of the outcomes of the three groups were done using the log-rank test.
The data are shown in FIG. 10A, and demonstrate that treatment of animals with the quinazolines of the invention (HI-P353 and HI-P364) inhibited the growth of breast cancer cell tumors as compared with untreated controls. [0132]
Inhibition of Brain Tumor Cells [0133]
An analogous experiment was done implanting brain tumor cells into mice. The right hind legs of CB.17 SCID mice were inoculated subcutaneously with 1×10[0134] ⁶of U373 human glioblastoma cells in 0.1 ml of PBS. Twenty four hours after inoculation, mice were treated with HI-P353 (10 mg/kg/day×5 days/week, N=7), or HI-P364 (10 mg/kg/day×5 days/week, N=8), or vehicle (50% DMSO in PBS, N=7) for four weeks. Mice were monitored daily for health status and tumor growth. Tumors were measured 3 times a week using a Vernier caliper. Tumor volumes were calculated using the following formula: (width)²×(length/2). Comparisons of outcome between groups were done using the log-rank test.
The data are shown in FIG. 10B and demonstrate that treatment of animals with the quinazolines of the invention (HI-P353 and HI-P364) inhibited the growth of brain tumors as compared with untreated controls. [0135]
Inhibition of Intracranial Brain Tumors [0136]
The anti-tumor activity of the quinazolines of the invention was also studied with intracranial tumors. Nude mice were first anesthetized with Avertin. Under aseptic conditions in a laminar flow hood, a small hole was drilled at 2 mm to the right of the midline and 2 mm posterior to the bregma. An amount of 4×10[0137] ⁵U87 glioblastoma cells in 10 μL of PBS were intracranially implanted using a Hamilton syringe into the right cerebral hemisphere of mice and a stereotaxic apparatus according to the method described in Huang, H. J. S. et al., J. Biol. Chem. 272:2927-2935, 1997.
Twenty-four hours after inoculation, mice were treated with 1H—P353 (20 mg/kg/day×10 days, n=10), HI-P364 (20 mg/kg/day×10 days, n=10), or vehicle (50% DMSO in PBS, n=10). The mice were monitored twice daily for health status. [0138]
FIG. 10C shows the survival rate of mice inflicted with intracranial tumors. Treatment of mice with quinazolines (HI-P353 and HI-P364) resulted in prolonged survival as compared with mice treated with vehicle alone. [0139]
All publications, patents, and patent documents described herein are incorporated by reference as if fully set forth. The invention described herein may be modified to include alternative embodiments. All such obvious alternatives are within the spirit and scope of the invention, as claimed below. [0140]

Claims

We claim:

1. A compound of the formula:

wherein:

R^ais iodo, (C₁-C₄)hydroxyalkyl, methylenedioxy, ethylenedioxy, benzyloxy, OCF₃. SCF₃SO₃H, SO₂F, SO₂NR²R³where R²is hydrogen or (C₁-C₄)alkyl and R³is hydrogen, (C_1-C₄)alkyl, or phenyl, NR²R⁴where R²is hydrogen or (C₁-C₄)alkyl and R⁴is phenyl; or a group of the formula

wherein R⁵and R⁶are each independently, hydrogen, (C₁-C₄)alkyl, or (C₁-C₄) perfluoroalkyl, and R⁷is hydrogen, halo, hydroxy, (C₁-C₄)alkyl, (C₁-C₄)alkoxy, (C₁-C₄)hydroxyalkyl, or N(R²)₂, where R²is hydrogen or (C₁-C₄)alkyl;

n is an integer of 1-4;

R^bis each, independently, hydrogen, halo, hydroxy, mercapto, (C₁-C₄)alkyl, (C₁-C₄)alkoxy, (C₁-C₄)thioalkyl, (C₁-C₄)hydroxyalkyl, nitro, cyano, methylenedioxy, ethylenedioxy, COCH_3,CF_3,OCF_3,SCF_3,COOH, SO₃H, SO₂F, phenyl, or phenyl substituted by a group selected from halo, hydroxy, mercapto, (C₁-C₄)alkyl, (C₁-C₄) alkoxy, (C₁-C₄)thioalkyl, (C₁-C₄)hydroxyalkyl, amino, nitro, cyano, CF₃, COOH, SO₃H, SO₂NR²R³, SO₂F where R²is H or (C₁-C₄)alkyl and R³is H, (C₁-C₄)alkyl, phenyl, or phenyl substituted by a group as defined above; benzyloxy or benzyloxy substituted on the phenyl portion by a group defined above; NR²R³where R²is H or (C₁-C₄)alkyl and R³is H, (C₁-C₄)alkyl, phenyl, or phenyl substituted by a group as defined above; and

R¹is (C₁-C₄)alkyl or a pharmaceutically acceptable salt thereof.

2. The compound of claim 1 wherein the pharmaceutically acceptable salt is an acid addition salt.

3. The compound of claim 1, wherein R¹is methyl.

4. The compound of claim 3, wherein R^ais a member selected from the group consisting of I, NHC₆H₅, —OCH₂CH₂O—, —OCH₂O—, OCF₃, SCF₃, CH₂OH, C₂H₄OH, SO₃H, SO₂NH₂, and SO₂F.

5. The compound of claim 4, wherein R^ais I, OCF_3,or SO₂F.

6. The compound of claim 5, wherein R^ais I.

7. The compound of claim 5 wherein R^ais OCF₃.

8. The compound of claim 3, wherein n is I and R^ais a group of the formula

9. The compound of claim 8, wherein R⁵and R⁶are each, independently, H, CH_3,or CF₃.

10. The compound of claim 9, wherein R⁵and R⁶are CF₃and R⁷is NH₂.

11. The compound of claim 3, wherein R^bis at least one member selected from the group consisting of F, Cl, Br, I, OH, NH₂, NO₂, CN, COOH, CH₃and CF₃.

12. The compound of claim 11, wherein R^bis at least one member selected from the group consisting of F, Cl, Br, OH and CF₃.

13. A pharmaceutical composition comprising a therapeutically effective amount of a compound of claim 1 and a pharmaceutically acceptable carrier or diluent.

14. A method of treating tumors in a subject comprising administering to said subject an anti-tumor effective amount of a compound of claim 1.

15. A method of treating leukemia in a subject comprising administering to said subject a compound of claim 1 in an amount effective for inducing apoptosis of leukemia cells.

16. A method of inhibiting cancer cell growth in a subject comprising administering to said subject a cytotoxic effective amount of a compound of claim 1.

17. A method of treating leukemia in a subject comprising administering to said subject a compound selected from the group consisting of:

4-(3′,5′-dibromo-4′-methylphenyl)amino-6,7-dimethoxyquinazoline,

4-(2′,4′,6′-tribromophenyl)amino-6,7-dimethoxyquinazoline,

4-(2′,3′,5′,6′-tetrafluoro-4′-bromophenyl)amino-6,7-dimethoxyquinazoline,

4-(4′-fluorophenyl)amino-6,7-dimethoxyquinazoline,

4-(3′-fluorophenyl)amino-6,7-dimethoxyquinazoline,

4-(2′-fluorophenyl)amino-6,7-dimethoxyquinazoline,

4-(4′-trifluoromethylphenyl)amino-6,7-dimethoxyquinazoline,

4-(2′-trifluoromethylphenyl)amino-6,7-dimethoxyquinazoline,

4-(3′,5′-bis-trifluoromethylphenyl)amino-6,7-dimethoxyquinazoline,

and pharmaceutically acceptable acid addition salts thereof.

18. A method of treating breast tumors in a subject comprising administering to said subject a compound selected from the group consisting of

4-(3′-bromophenyl)amino-6,7-dimethoxyquinazoline,

4-(3′,5′-dibromo-4′-hydroxyphenyl)amino-6,7-dimethoxyquinazoline,

4-(3′-chloro-4′-hydroxyphenyl)amino-6,7-dimethoxyquinazoline,

4-(3′,5′-bis-trifluoromethylphenyl)amino-6,7-dimethoxyquinazoline,

4-(4′-fluorophenyl)amino-6,7-dimethoxyquinazoline,

4-(3′-fluorophenyl)amino-6,7-dimethoxyquinazoline,

4-(2′-fluorophenyl)amino-6,7-dimethoxyquinazoline, and

pharmaceutically acceptable acid addition salts thereof.

19. A method of treating multi-drug resistant leukemia in a subject comprising administering to said subject 4-(4′-hydroxyphenyl)amino-6,7-dimethoxyquinazoline or a pharmaceutically acceptable salt thereof, in an amount effective for inducing apoptosis of multi-drug resistant leukemia cells.

20. A method of treating multi-drug resistant leukemia in a subject comprising administering to said subject 4-(3′-bromo-4′-hydroxyphenyl)amino-6,7-dimethoxyquinazoline or a pharmaceutically acceptable acid addition salt thereof, in an amount effective for inducing apoptosis of multi-drug resistant leukemia cells.

21. The compound 4-(3′,5′-dibromo-4′-methylphenyl)amino-6,7-dimethoxyquinazoline or a pharmaceutically acceptable acid addition salt thereof.

22. The compound 4-2′,4′,6′-tribromophenyl)amino6,7-dimethoxyquinazoline or a pharmaceutically acceptable acid addition salt thereof.

23. The compound 4-(2′,3′,5′,6′-tetrafluoro-5′-bromophenyl)amino-6,7-dimethoxyquinazoline or a pharmaceutically acceptable acid addition salt thereof.

24. The compound 4-(4′-fluorophenyl)amino-6,7-dimethoxyquinazoline or a pharmaceutically acceptable acid addition salt thereof.

25. The compound 4-(4′-trifluoromethylphenyl)amino-6,7-dimethoxyquinazoline or a pharmaceutically acceptable acid addition salt thereof.

26. The compound 4-(3′,5′-bis-trifluoromethylphenyl)amino-6,7-dimethoxy-quinazoline or a pharmaceutically acceptable acid addition salt thereof.

27. A pharmaceutical composition comprising an effective amount of a compound selected from the group consisting of:

4-(3,5-bis-trifluoromethylphenyl)amino-6,7-dimethoxyquinazoline,

4-(2′,4′,6′-tribromophenyl)amino-6,7-dimethoxyquinazoline,

4-(3′,5′-dibromo-4′-methylphenyl)amino-6,7-dimethoxyquinazoline,

4-(4′-fluorophenyl)amino-6,7-dimethoxyquinazoline,

4-(4′-tifluoromethylphenyl)amino-6,7-dimethoxyquinazoline, and

pharmaceutically acceptable acid addition salts thereof; and

a pharmaceutically acceptable carrier or diluent.