WO1995012583A1 - New quinazolines as inhibitors of hiv reverse transcriptase - Google Patents

Abstract

Compounds having a quinazolin-2-one nucleus with a substituted alkynyl or substituted alkenyl at the 4-position are described. These compounds are useful in the inhibition of HIV reverse transcriptase (including its resistant varieties), the prevention or treatment of infection by HIV and the treatment of AIDS, either as compounds, pharmaceutically acceptable salts, pharmaceutical composition ingredients, whether or not in combination with other antivirals, immunomodulators, antibiotics or vaccines. Methods of treating AIDS and methods of preventing or treating infection by HIV are also described.

Description

TITLE OF THE INVENTION

NEW QUINAZOLINES AS INHIBITORS OF HIV REVERSE

TRANSCRIPTASE

BACKGROUND OF THE INVENTION

This application is a continuation-in-part of Merck Case 18727IA, USSN 07/991,164, filed December 16,1992, which is a continuation-in-part-of Merck Case 18727, USSN 07/880,119, filed May 7, 1992.

A retrovirus designated human immunodeficiency virus (HIV) is the etiological agent of the complex disease that includes progressive destruction of the immune system (acquired immune deficiency syndrome; AIDS) and degeneration of the central and peripheral nervous system. This virus was previously known as LAV, HTLV-III, or ARV. A common feature of retrovirus replication is reverse transcription of the RN A genome by a virally encoded reverse transcriptase to generate DNA copies of HIV sequences, a required step in viral replication. It is known that some compounds are reverse transcriptase inhibitors and are effective agents in the treatment of AIDS and similar diseases, e.g., azidothymidine or AZT.

Nucleotide sequencing of HIV shows the presence of a pol gene in one open reading frame [Ratner, L. et al, Nature, 313, 277(1985)]. Amino acid sequence homology provides evidence that the pol sequence encodes reverse transcriptase, an endonuclease and an HIV protease [Toh, H. et al, EMBO J., 4, 1267 (1985); Power, M.D. et al, Science, 231, 1567 (1986); Pearl, L.H. et al, Nature, 329, 351 (1987)].

Applicants demonstrate that the compounds of this invention are inhibitors of HIV reverse transcriptase. The particular advantages of the present compounds are their demonstrated inhibition of resistant HIV reverse transcriptase. BRIEF DESCRIPTION OF THE INVENTION

Compounds of Formula I, as herein defined, are disclosed. These compounds are useful in the inhibition of HIV reverse transcriptase (and its resistant varieties), the prevention of infection by HIV, the treatment of infection by HIV and in the treatment of AIDS and/or ARC, either as compounds, pharmaceutically acceptable salts (when appropriate), pharmaceutical composition ingredients, whether or not in combination with other antivirals, anti-infectives, immunomodulators, antibiotics or vaccines. Methods of treating AIDS, methods of preventing infection by HIV, and methods of treating infection by HIV are also disclosed.

DETAILED DESCRIPTION OF THE INVENTION AND PREFERRED

EMBODIMENTS

This invention is concerned with compounds of Formula I, combinations thereof, or pharmaceutically acceptable salts thereof, in the inhibition of HIV reverse transcriptase and its resistant varieties, the prevention or treatment of infection by HIV and in the treatment of the resulting acquired immune deficiency syndrome (AIDS). Compounds of Formula I are defined as follows:

I wherein X is O;

G, when present is halo, nitro, or cyano; n is 0-4;

Rl is C3-5cycloalkyl, C2-5alkynyl, C2-4alkenyl, or cyano;

R2 is C2-5alkynyl substituted with one or more of A, or C2-5alkenyl substituted with one or more of A, wherein A is i) halo; ii) hydroxy; iii) amino; iv) cyano; v) nitro; vi) azido; vii) C3-8cycloalkyl; viii) Cl-4alkoxy, unsubstituted or substituted with one or more of halo; ix) di-(Cl-4alkyl)amino; x) Cl-4alkylamino; xi) aryl, unsubstituted or substituted with one or more of D, wherein D is amino, nitro, cyano, or Cl-3alkoxy; xii) aryloxy, unsubstituted or substituted with one or more of D; xiii) heterocycle, unsubstituted or substituted with one or more of

D; xiv) heterocycle oxy; or xv) C2-5alkenyl; xvi) COOR, wherein R is H, Cl-4alkyl or aryl; xvii) CONR2; or xviii) COR;

R3 i IS i) H; ii) cyano; iii) amino; iv) hydroxyl; v) Cl -4alkyl, unsubstituted or substituted with one or more of

E, wherein E is halo, hydroxyl, amino, nitro, cyano,

Cl-4alkoxy, or C3-5cycloalkyl; vi) C2-4alkenyl, unsubstituted or substituted with E; or vii) C2-4alkynyl, unsubstituted or substituted with E; R4 is i) H; ii) Cl-4alkyl; iii) Cl-5alkylcarbonyl; iv) benzoyl, unsubstituted or substituted with one or more of A; or v) heterocyclecarbonyl; with the proviso that any terminal alkynyl carbon is not substituted with any substituent selected from the group consisting of halo, hydroxy, amino, cyano, nitro, azido, Cl-4alkoxy unsubstituted or substituted with one or more of halo, di-(Cl-4alkyl)amino, Cl-4alkylamino, aryloxy unsubstituted or substituted with one or more of D, or heterocycleoxy; or a pharmaceutically acceptable salt thereof.

In one embodiment, compounds further are limited to Formula II:

π wherein:

R2 is C2-5 alkynyl substituted with halo, hydroxy, amino, cyano, nitro, azido, C3-8 cycloalkyl, Cl-4 alkoxy, di-(Cl-4alkyl)- amino, Cl-4alkylamino, phenyl, 2-nitrophenyl, pyridyl, pyrimidyl, pyrazinyl, imidazolyl, or C2-3alkenyl;

R³ is H or Cl-3 alkyl; or a pharmaceutically acceptable salt thereof.

Specific illustrations of the compounds of this invention include those of the following Table. TABLE

Compound Ex. R3 R ] [C™ IC oDBL Mutant

1 4 H CH₂OCH₃ 49 nM 7300 nM

2 3 CH₃ CH₂OCH₃ 13 nM 700 nM

3 20 CH₃ CH₂OCH₂CF₃ 19 nM 2600 nM

4 13 CH₃ CH₂CH₂F 7 nM 230 nM

5 16 H CH₂CH₂F 31 nM 3500 nM

6 14 CH₃ CH₂CH₂C1 7 nM 250 nM

7 18 CH₃ CH₂N₃ 20 nM 1750 nM

8 17 CH₃ CH₂F 24 nM 2900 nM

9 33 CH₃ CH₂N(CH₃)₂ 54 nM 273 nM

10 29 CH₃ 2-pyridyl 21 nM 87 nM

11 24 H 2-pyridyl (+/-) 19 nM 635 nM

12 25 H 3-pyridyl 84 nM 310 nM

13 26 H 4-pyridyl 470 nM 3000 nM

14 28 H 5-pyrimidyl 1750 nM 3000 nM

15 35 H 2-pyrimidyl 130 nM 8800 nM

16 27 H 2-pyrazinyl 270 nM 9000 nM

17 21 CH₃ CH₂0-4- 165 nM 3900 nM pyridyl

18 22 CH₃ CH₂0-4- 9400 nM 65000 nM pyridyl-N-oxide

19 9 CH₃ CH₂(4- 1000 nM 22000 nM moφholinyl) Compound Ex. R3 R IC n ICsoDBL Mutant

20 19 CH₃ CH₂(1- 620 nM 960 nM imidazolyl)

21 35 CH₃ CH=CH₂ 4.3 nM 248 nM

22 36 CH₃ CH₂OH 255 nM

23 34 H phenyl 6.1 nM 135 nM

24 37 CH₃ CH₂0-2- 125 nM 4900 nM pyridyl

25 38 H 2-nitrophenyl 10.5 nM 390 nM

26 40 H 2-pyridyl (-) 7 nM 400 nM

Preferred compounds include:

6-chloro-4-cyclopropyl-4-(4-fluoro-l-butynyl)-3, 4-dihydro-3-methyl- quinazolin-2(lH)-one (Compound 4),

6-chloro-4-cyclopropyl-3,4-dihydro-3-methyl-4-((2-pyridyl)ethynyl)- quinazolin-2(lH)-one (Compound 10),

6-chloro-4-cyclopropyl-3,4-dihydro-4-((2-pyridyl)ethynyl)quinazolin- 2(lH)-one (Compound 11),

6-chloro-4-cyclopropyl-3,4-dihydro-4-(phenylethynyl)quinazolin-2-(lH)- one (Compound 23), or

(-)6-chloro-4-cyclopropyl-3,4-dihydro-4-((2-pyridyl)ethynyl)-quinazolin-

2(lH)-one (Compound 26), or a pharmaceutically acceptable salt thereof.

Compound 26 has (S) stereochemistry at the 4-position, with the structure:

6-chloro-4(S)-cyclopropyl-3,4-dihydro-4-((2-pyridyl)ethynyl)quin- azolin-2(lH)-one. This compound has also been isolated as a monohydrate.

The compounds of the present invention may have asymmetric centers and may occur, except when specifically noted, as racemates, racemic mixtures or as individual diastereomers, or enantiomers, with all isomeric forms being included in the present invention.

When any variable (e.g., G, Rl, R2, R3, etc.) occurs more than one time in any constituent or in Formula I, its definition on each occurrence is independent of its definition at every other occurrence. Also, combinations of substituents and/or variables are permissible only if such combinations result in stable compounds.

As used herein except where noted, "alkyl" is intended to include both branched- and straight-chain saturated aliphatic hydrocarbon groups having the specified number of carbon atoms; "alkenyl" is intended to cover both branched- and straight chain alkyl groups with at least one carbon-carbon double bond; "alkynyl" is intended to cover both branched- and straight chain alkyl groups with at least one carbon-carbon triple bond. "Halogen" or "halo" as used herein, means fluoro, chloro, bromo and iodo.

As used herein, with exceptions as noted, "aryl" is intended to mean phenyl, naphthyl, tetrahydronaphthyl, biphenyl, phenanthryl, anthryl or acenaphthyl.

The term heterocycle or heterocyclic, as used herein except where noted, represents a stable 5- to 7-membered monocyclic or stable 8- to 11-membered bicyclic heterocyclic ring which is either saturated, partially unsaturated or unsaturated, and which consists of carbon atoms and from one to four heteroatoms selected from the group consisting of N, O and S, and wherein the nitrogen and sulfur heteroatoms may optionally be oxidized, and including any bicyclic group in which any of the above-defined heterocyclic rings is fused to a benzene ring. The heterocyclic ring may be attached at any heteroatom or carbon atom which results in the creation of a stable structure. Examples of such heterocyclic elements include piperidinyl, piperazinyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, 2-oxoazepinyl, azepinyl, pyrrolyl, 4- piperidonyl, pyrrolidinyl, pyrazolyl, pyrazolidinyl, imidazolyl, imidazolinyl, imidazolidinyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, oxazolyl, oxazolidinyl, isoxazolyl, isoxazolidinyl, moφholinyl, thiazolyl, thiazolidinyl, isothiazolyl, quinuclidinyl, isothiazolidinyl, indolyl, quinolinyl, isoquinolinyl, benzimidazolyl, thiadiazoyl, benzopyranyl, benzothiazolyl, benzoxazolyl, furyl, tetrahydrofuryl, benzofuranyl, tetrahydropyranyl, thienyl, benzothienyl, thiamoφholinyl, thiamoφholinyl sulfoxide, thiamoφholinyl sulfone, and oxadiazolyl.

The compounds of the present invention can be synthesized by the following methods.

METHOD A

5a

The characteristic feature of Method A is R-metal addition to a dihydroquinazoline in the presence of magnesium ions and other good Lewis acids. Method A is further illustrated by Examples 24-29. METHOD B

B), H

8

Method B involves a cross-coupling reaction in the presence of palladium (II) chloridetriphenylphosphine couplex as a catalyst, to give aryl and heterocyclic substitutions of the 4-acetylene group. Example 35 illustrates the Method. METHOD C

5a Method C depicts another method of obtaining substituted 4- acetylene derivatives. A tetrahydropyran derivative 9 is formed by R- metal addition as in Method A, followed by reaction with an alcohol in the presence of pyridinium paratoluene sulf onate (PPTS) to form the corresponding alcohol intermediate ii, wherein R is CH₃. Chlorination, followed by nucleophilic substitution with the desired end group gives the appropriate product 5a. Method C is specifically illustrated by Examples 5-9.

METHOD D

9 EtOH/PPTS

11

Method D is suitable for halo substituted 4-alkynyl derivatives. The penultimate hydroxy derivative ϋ is formed as in Method C, followed by reaction with the florinating agent diethylamino- sulfurtrifluoride (DAST). Deprotection may then be desired. Chlorination is a side reaction. Method D is specifically illustrated by Examples 11-14.

The compounds of the present inventions are useful in the inhibition of HIV reverse transcriptase, the prevention of treatment of infection by human immunodeficiency virus (HIV) and the treatment of consequent pathological conditions such as AIDS. Treating AIDS or preventing or treating infection by HTV is defined as including, but not limited to, treating a wide range of states of HIV infection: AIDS, ARC (AIDS related complex), both symptomatic and asymptomatic, and actual or potential exposure to HIV. For example, the compounds of this invention are useful in treating infection by HIV after suspected Oast exposure to HTV by e.g., blood transfusion, exchange of body fluids, bites, accidental needle stick, or exposure to patient blood durir ■^■? surgery.

The particular advantage of the compounds of this invention is their potent inhibition against HTV .-.verse transcriptase rendered resistant to other antivirals, such as L-697,661, which is 3-([(4,7- dichloro- 1 ,3-benzoxazol-2-yl)methyl]-amino)-5-ethyl-6-methyl-pyridin- 2(lH)-one; or L-696,229, which is 3-[2-(l ,3-benzoxazol-2-yl)ethyl]-5- ethyl-6-methylpyridin-2(lH)-one; or AZT.

The compounds of this invention are also useful in the preparation and execution of screening assays for antiviral compounds. For example, the compounds of this invention are useful for isolating enzyme mutants, which are excellent screening tools for more powerful antiviral compounds. Furthermore, the compounds of this invention are useful in establishing or determining the binding site of other antivirals to HTV reverse transcriptase, e.g., by competitive inhibition. Thus the compounds of this invention are commercial products to be sold for these puφoses.

For these puφoses, the compounds of the present invention may be administered orally, parenterally (including subcutaneous injections, intravenous, intramuscular, intrastemal injection or infusion techniques), by inhalation spray, or rectally, in dosage unit formulations containing conventional non-toxic pharmaceutically-acceptable carriers, adjuvants and vehicles.

Thus, in accordance with the present invention there is further provided a method of treating and a pharmaceutical composition for treating HIV infection and AIDS. The treatment involves administering to a patient in need of such treatment a pharmaceutical composition comprising a pharmaceutical carrier and a therapeutically- effective amount of a compound of the present invention.

These pharmaceutical compositions may be in the form of orally-administrable suspensions or tablets; nasal sprays; sterile injectable preparations, for example, as sterile injectable aqueous or oleagenous suspensions or suppositories.

When administered orally as a suspension, these compositions are prepared according to techniques well-known in the art of pharmaceutical formulation and may contain microcrystalline cellulose for imparting bulk, alginic acid or sodium alginate as a suspending agent, methylcellulose as a viscosity enhancer, and sweeteners/flavoring agents known in the art. As immediate release tablets, these compositions may contain microcrystalline cellulose, dicalcium phosphate, starch, magnesium stearate and lactose and/or other excipients, binders, extenders, disintegrants, diluents and lubricants known in the art.

When administered by nasal aerosol or inhalation, these compositions are prepared according to techniques well-known in the art of pharmaceutical formulation and may be prepared as solutions in saline, employing benzyl alcohol or other suitable preservatives, absoφtion promoters to enhance bioavailability, fluorocarbons, and/or other solubilizing or dispersing agents known in the art.

The injectable solutions or suspensions may be formulated according to known art, using suitable non-toxic, parenterally-acceptable diluents or solvents, such as mannitol, 1,3-butanediol, water, Ringer's solution or isotonic sodium chloride solution, or suitable dispersing or wetting and suspending agents, such as sterile, bland, fixed oils, including synthetic mono- or diglycerides, and fatty acids, including oleic acid. When rectally administered in the form of suppositories, these compositions may be pre! ared by mixing the drug with a suitable non-irritating excipient, such a& cocoa butter, synthetic glyceride esters or polyethylene glycols, which are solid at ordinary temperatures, but liquidify and/or dissolve in the rectal cavity to release the drag.

The compounds of this invention can be administered orally to humans in a dosage range of 1 to 100 mg/kg body weight in divided doses. One preferred dosage range is 0.1 to 10 mg/kg body weight orally in divided doses. Another preferred dosage range is 0.1 to 20 mg/kg body weight orally in divided doses. It will be understood, however, that the specific dose level and frequency of dosage for any particular patient may . varied and will depend upon a variety of factors including the activity of the specific compound employed, the metabolic stability and length of action of that compound, the age, body weight, general health, sex, diet, mode and time of administration, rate of excretion, drug combination, the severity of the particular condition, and the host undergoing therapy.

The present invention is also directed to combinations of the HIV reverse transcriptase inhibitor compounds with one or more agents useful in the treatment of AIDS. For example, the compounds of this invention may be effectively administered, whether at periods of pre- exposure and/or post-exposure, in combination with effective amounts of the AIDS antivirals, immunomodulators, a infectives, or vaccines, such as those in the following Table C.

TABLE C

Drug Name Manufacturer Indication

AL-721 Ethigen ARC, PGL

(Los Angeles, CA) HIV positive, AIDS

Recombinant Human Triton Biosciences AIDS, Kaposi's

Inteiferon Beta (Almeda, CA) sarcoma, ARC

Acemannan Caπϊngton Labs ARC

(Irving, TX) (See also immunomodulators)

Cytovene Syntex sight threatening CMV

Ganciclovir (Palo Alto, CA) peripheral CMV retinitis

d4T Bristol-Myers AIDS, ARC

Didehydrodeoxy- (New York, NY) thymidine

ddl Bristol-Myers AIDS, ARC

Dideoxyinosine (New York, NY)

EL10 Elan Coφ, PLC HIV infection

(Gainesville, GA) (See also immunomodulators) Drug Name Manufacturer Indication Trisodium Indication Astra Pharm CMV retinitis, HIV Phosphonoformate Products, Inc. infection, other CMV (Westborough, MA) infections

Dideoxycytidine; Hoffman-La Roche AIDS, ARC ddC (Nutley, NJ)

Novapren Novaferon Labs, Inc. HIV inhibitor (Akron, OH) Diapren, Inc. (Roseville, MN, marketer)

Peptide T Peninsula Labs AIDS

Octapeptide (Belmont, CA)

Sequence

Zidovudine; AZT Burroughs Wellcome AIDS, adv, ARC (Rsch. Triangle Park, pediatric AIDS

NC) Kaposi's sarcoma, asymptomatic HTV infection, less severe HIV disease, neurological involvement, in combination with other therapies. Drug Name Manufacturer Indication

Ansamycin LM 427 Adria Laboratories ARC (Dublin, OH) Erbamont (Stamford, CT)

Dextran Sulfate Ueno Fine Chem. AIDS, ARC, HIV

Ind. Ltd. positive asymptomatic

(Osaka, Japan)

Virazole Viratek/ICN asymptomatic HIV

Ribavirin (Costa Mesa, CA) positive, LAS, ARC

Alpha Interferon Burroughs Wellcome Kaposi's sarcoma, HIV

(Rsch. Triangle in combination

Park, NC) w/Retrovir

Acyclovir Burroughs Wellcome AIDS, ARC, asymptomatic HIV positive, in combination with AZT.

Antibody which Advanced Biotherapy AIDS, ARC neutralizes pH labile Concepts alpha aberrant (Rockville, MD)

Interferon in an immuno-adsoφtion column Drug Name Manuiacturer Indication L-697,661 Merck AIDS, ARC,

(Rahway, NJ) asymptomatic

HIV positive, also in combination with AZT.

L-696,229 Merck AIDS, ARC,

(Rahway, NJ) asymptomatic

HTV positive, also in combination with AZT.

L-735,524 Merck AIDS, ARC,

(Rahway, NJ) asymptomatic

HIV positive, also in combination with AZT.

IMMUNO-MODULATORS

Drug Name Manufacturer Indication AS-101 Wyeth-Ayerst Labs. AIDS

(Philadelphia, PA)

Bropirimine Upjohn advanced AIDS

(Kalamazoo, MI)

Acemannan Carrington Labs, Inc. AIDS, ARC

(Irving, TX) (See also antivirals) Drug Name Manufacturer Indication

CL246.738 American Cyanamid AIDS, Kaposi's

(Pearl River, NY) sarcoma

Lederle Labs

(Wayne, NJ)

EL10 Elan Coφ, PLC HIV infection

(Gainesville, GA) (See also antivirals)

Gamma Interferon Genentech ARC, in combination

(S. San Francisco, w/TNF (tumor

CA) necrosis factor)

Granulocyte Genetics Institute AIDS

Macrophage Colony (Cambridge, MA)

Stimulating Sandoz

Factor (East Hanover, NJ)

Granulocyte Hoeschst-Roussel AIDS

Macrophage Colony (SomerviUe, NJ)

Stimulating Immunex

Factor (Seattle, WA)

Granulocyte Schering-Plough AIDS

Macrophage Colony (Madison, NJ)

Stimulating Factor AIDS, in combination w/AZT

HIV Core Particle Rorer seropositive HIV

Immunostimulant (Ft. Washington, PA)

IL-2 Cetus AIDS, in combination

Interleukin-2 (Emeryville, CA) w/AZT Drug Name Manufacturer Indication

IL-2 Hoffman-La Roche AIDS, ARC, HIV, in

Interleukin-2 (Nutley, NJ) combination

Immunex w/AZT

Immune Globulin Cutter Biological pediatric AIDS, in

Intravenous (Berkeley, CA) combination

(human) w/AZT

IMREG- 1 Imreg AIDS, Kaposi's

(New Orleans, LA) sarcoma, ARC, PGL

IMREG-2 Imreg AIDS, Kaposi's

(New Orleans, LA) sarcoma, ARC, PGL

Imuthiol Diethyl Merieux Institute AIDS, ARC

Dithio Carbamate (Miami, FL)

Alpha-2 Schering Plough Kaposi's sarcoma

Interferon (Madison, NJ) w/AZT: AIDS

Methionine- TNI Pharmaceutical AIDS, ARC

Enkephalin (Chicago, IL)

MTP-PE Ciba-Geigy Coφ. Kaposi's sarcoma

Muramyl- (Summit, NJ)

Tripeptide

Granulocyte Amgen AIDS, in combinatioi

Colony (Thousand Oaks, CA) w/AZT

Stimulating

Factor Drug Name Manufacturer Indication rCD4 Genentech AIDS, ARC

Recombinant (S. San Francisco,

Soluble Human CD4 CA)

rCD4-IgG AIDS, ARC hybrids

Recombinant Biogen AIDS, ARC

Soluble Human CD4 (Cambridge, MA)

Interferon Hoffman-La Roche Kaposi's sarcoma

Alfa 2a (Nutley, NJ) AIDS, ARC, in combination w/AZT

SK&F106528 Smith, Kline & French HIV infection

Soluble T4 Laboratories (Philadelphia, PA)

Thymopentin Immunobiology HIV infection Research Institute (Annandale, NJ)

Tumor Necrosis Genentech ARC, in combina

Factor; T F (S. San Francisco, tion w/gamma

CA) Interferon

ANTI-INFECTIVES

Drug Name Manufacturer Indication Clindamycin with Upjohn PCP Primaquine (Kalamazoo, MI) Drug Name Manufacturer Indication Fluconazole Pfizer cryptococcal

(New York, NY) meningitis, candidiasis

Pastille Squibb Coφ. prevention of Nystatin Pastille (Princeton, NJ) oral candidiasis

Ornidyl Merrell Dow PCP Eflornithine (Cincinnati, OH)

Pentamidine LyphoMed PCP treatment Isethionate (IM (Rosemont, IL) & IV)

Piritrexim Burroughs Wellcome PCP treatment (Rsch. Triangle Park, NC)

Pentamidine Fisons Coφoration PCP prophylaxis isethionate for (Bedford, MA) inhalation

Spiramycin Rhone-Poulenc cryptosporidial Pharmaceuticals diarrhea (Princeton, NJ)

Intraconazole- Janssen Pharm histoplasmosis; R51211 (Piscataway, NJ) cryptococcal meningitis

Trimetrexate Warner-Lambert PCP - "t -

OTHER

Drug Name Manufacturer Indication

Recombinant Human Ortho Pharm. Coφ. severe anemia

Erythropoietin (Raritan, NJ) assoc. with AZT therapy

Megestrol Acetate Bristol-Myers treatment of

(New York, NY) anorexia assoc. w/AIDS

Total Enteral Norwich Eaton diarrhea and

Nutrition Pharmaceuticals malabsoφtion

(Norwich, NY) related to AIDS

It will be understood that the scope of combinations of the compounds of this invention with AIDS antivirals, immunomodulators, anti-infectives or vaccines is not limited to the list in the above Table, but includes in principle any combination with any pharmaceutical composition useful for the treatment of AIDS. The compound L-735,524 is an HIV protease inhibitor with the chemical name N-(2(R)-hydroxy- 1 (S)-indanyl)-2(R)-phenylmethyl-4-(S)-hydroxy-5-(l -(4-(3-pyridyl- methyl)-2(S)-N'-(t-butylcarboxamido)-piperazinyl))-pentaneamide, or pharmaceutically acceptable salt thereof. L-735,524 is synthesized by the methods of EP 0541168, herein incoφorated by reference for these puφoses. EXAMPLE 1

6-chloro-4-cyclopropyl-3,4-dihydro-l-(4-methoxybenzyl)-4-(3-methoxy- 1 -propynyl quinazolin-2π H)-one

A. (2-Amino-4-chlorophenyl cyclopropyl ketone

A solution of cyclopropylmagnesium bromide, prepared from 2.4 g (0.099 g atom) of magnesium turnings and 13.0 g (0.107 mol) of cyclopropyl bromide in 100 mL of THF, was stirred at 38 °C as a solution of 5-chloro-anthranilonitrile (3.65 g, 0.0239 mol) in 40 mL THF was added over 20 minutes. Stirring was continued at 40°C for 2 h, following which the reaction mixture was cooled in an ice bath and 50 mL of saturated NH4CI was added, followed by 100 mL of 2N HCl. The cooling bath was removed and stirring was continued at room temperature for 2 h. The mixtu -^■ was then brought to pH 9 by addition of 20% NaOH and was extracted . imes with ether. The combined organic phases were washed with brine and dried over MgS04. Following removal of the solvents the oily residue was flash chromatographed, eluting with 15% EtOAc in hexane, to provide 3.0 g (64%) of the title compound as a yellow solid, mp 66-68°C; iH NMR (CDCI3): δ 1.02 (m, 2H), 1.18 (m, 2H), 2.57 (m, 1H), 6.17 (s, 1H), 6.61 (d, J=8.5 Hz, IH), 7.1.2 (dd, J=2, 8.5 Hz, IH), 7.92 (d, J=2 Hz, IH).

B. 6-chloro-4-cyclopropylquinazolin-2(THVone

To a stirred suspension of 150 mg (0.767 mmol) of the product from Step A in 3 mL of glacial acetic acid at 0° was added a solui 1 of 75 mg (0.922 mmol) of potassium cyanate in 0.3 mL of H2θ in on^ portion. After stirring for 1 hour at 0°-5°, the reaction mixture was allowed to warm to room temperature over a 1 hour period. The reaction mixture was partitioned between EtOAc and H2O, the organic layer washed with H2O, filtered, washed with brine, dried over Na2Sθ4, and concentrated to afford 120 mg of a light yellow solid. This material was chromatographed on silica gel to give 122 mg of the title compound as a solid:

!H NMR (CDC13): δ 1.27 (m, 2H), 1.57 (m, 2H), 2.55 (m, IH), 7.48 (d,

J=8 Hz, IH), 7.61 (dd, J=8, 2 Hz, IH), 8.10 (d, J=2 Hz, IH). FAB MS

M+H=221, mp=215-217°C.

C. 6-chloro-4-cyclopropyl- 1 -(4-methoxybenzyl)quinazolin-

2(THVone

To a stirred solution of 75 mg (0.338 mmol) of the product from Step B in 6 mL of dry DMF was added 17 mg (0.423 mmol) of sodium hydride (60% in mineral oil) in one portion. After 20 minutes when gas evolution ceased, 50 μL (0.372 mmol) of 4-methoxybenzyl- chloride was added in one portion. The reaction solution was stirred at room temperature for 2.5 hours, then heated to 80° under Ar for 4 hours, and allowed to stir at room temperature for 2.5 days. The reaction mixture was concentrated at reduced pressure and the residue partitioned between EtOAc and H2θ. The organic layer was washed with water, brine, dried over Na2Sθ4, and concentrated to give a residue which was chromatographed on silica gel using 1:1 EtOAc-hexane to give 73 mg of the title compound. An analytical sample was obtained by crystallization from EtOAc-hexane: iH NMR (CDCI3): δ 1.25 (m, 2H), 1.56 (m, 2H), 2.52 (m, IH), 3.76 (s, 3H), 5.40 (s, 2H), 6.83 (d, J=8.7 Hz, 2H), 7.18 (d, J=8.7 Hz, 2H), 7.21 (d, J=9.2 Hz, IH), 7.53 (dd, J=2.3, 9.2 Hz, IH), 8.1 (d, J=2.3Hz, IH). FAB MS M+H=341, mp 211-213°C.

D. 6-chloro-4-cyclopropyl-3,4-dihydro-l -^-methoxyenzy ^- O-methoxy-l-propynyDquinazolin^πHVone

A suspension of 548 mg (1.61 mmol) of 6-chloro-4- cyclopropyl-l-(4-methyoxybenzyl)-quinazolin-2(lH)-one and 1.55 g (4.82 mmol) of magnesium triflate was stirred for 30 minutes in 18 mL of ether under Ar at room temperature. To this solution was added a -78° solution of l-lithio-3-methoxypropyne (prepared by adding 543 μL (6.4 mmol) of 3-methoxy-l-propyne dropwise to a -78° solution of 6.4 mmol of lithiumdiisoproylamide in 18 mL of THF under Ar) via cannula. After 2 h at rt, the reaction was quenched by pouring into ice-cold IM citric acid and extracted with two portions of CHCI3. The organic layers were washed with 10% Na2Cθ3, dried over MgSOφ, treated with activated carbon and solvents removed to give 662 mg of an amber foam: NMR (CDCI3): δ 0.55-0.72 (m, 2H), 0.74-0.90 (m, 2H), 1.38-1.50 (m, IH), 3.32 (s, 3H), 3.76 (s, 3H), 4.08 (s, 2H), 5.02 (d, J=16.8 Hz, IH), 5.19 (d, J=16.8 Hz, IH), 5.36 (s, IH), 6.75 (d, J=9.0 Hz, IH), 6.82 (d, J=9.0 Hz, 2H), 7.12 (dd, J=9.0, 2.4 Hz, IH), 7.18 (d, J=9.0 Hz, 2H), 7.49 (d, J=2.4 Hz, IH).

EXAMPLE 2

6-ch]oro-4-cyclopropyl-3,4-dihydro-3-methyl-l-(4-methoxybenzyl)-4-

(3 -methoxy- 1 -propyny l)quinazolin-2( 1 HV one

A quantity of 130 mg (0.316 mmol) of the product from Example 1 was dissolved in 5 mL of dry DMF and treated with 25 mg (0.63 mmol) of 60% sodium hydride in oil under Ar. After stirring at room temperature for 40 minutes, 40 μL (0.63 mmol) of methyl iodide (dried by passing through a pad of alumina) was added in one portion via syringe, and the reaction mixture stirred overnight at room temperature. The DMF was removed by rotovap and the residue partitioned between CHCI3 and IM citric acid. The aqueous layer was extracted with CHCI3, and the combined organic layers washed with 10% Na2C03, dried over MgSθ4, and solvents removed to afford 140 mg of a dark brown oil which was chromatographed on 13 g of fine Siθ2 using 98.5:1.5 to 90:10 CHCI3-CH3CN to give 49 mg of the title compound as a pale yellow oil: NMR (CDCI3): δ 0.40-0.55 (m, 2H), 0.66-0.90 (m, 2H), 1.25-1.40 (m, IH), 3.29 (s, 3H), 3.40 (s, 3H), 3.76 (s, 3H), 4.21 (s, 2H), 4.98 (d, J=16.8 Hz, IH), 5.21 (d, J=16.8 Hz, IH), 6.70 (d, J=8.8 Hz, IH), 6.83 (d, J=8.8 Hz, 2H), 7.10 (dd, J=8.8, 2.4 Hz, IH), 7.165 (d, J=8.8 Hz, 2H), 7.47 (d, J=2.4 Hz, IH). EXAMPLE 3

6-chloro-4-cyclopropyl-3,4-dihydro-3-memyl-4-(3-methoxy-l- propynyl quinazolin-2(l H)-one

A quantity of 49 mg (0.115 mmol) of the product from Example 2 was treated with a solution of 2.5 mL of trifluoroacetic acid in 3.5 mL of methylene chloride for 3 h under Ar. The solvents were removed by rotary evaporation and the residue partitioned between CHC13 and 10% Na2Cθ3. The organic layer was dried over Na2S04 and the solvents removed to give 47 mg of a yellow oil which was chromatographed on 5 g fine Siθ2 using 98:2 CHCI3-CH3OH to afford 33 mg of the title compound which solidified upon lyophilization from dioxane: mp 119-121 °C,

NMR (CDCI3): δ 0.40-0.55 (m, 2H), 0.66-0.87 (m, 2H), 1.32-1.42 (m, IH), 3.25 (s, 3H), 3.41 (s, 3H), 4.21 (s, 2H), 6.77 (d, J=8.5 Hz, IH), 7.17 (dd, J=8.5, 2.3 Hz, IH), 7.38 (d, J=2.3 Hz, IH), 9.23 (s, IH). Anal. Calc'd for Cl6HπClN2θ2«0.5 H2O

C, 61.24; H, 5.78; N, 8.92 Found C, 61.07; H, 5.47; N, 8.63

EXAMPLE 4

6-chloro-4-cyclopropyl-3,4-dihydro-4-(3-methoxy-l-propynyl)- quinazolin-2( 1 HVone

A quantity of 48 mg (0.116 mmol) of the product from Example 1 was treated according to the procedure of Example 3 above for 21 h to give a yellow oil which was chromatographed on 7 g fine Siθ2 using 97:3 CHCI3-CH3OH to afford 12 mg of the title compound as an amoφhous solid upon lyophilization from dioxane: NMR (CDCI3): δ 0.57-0.68 (m, 2H), 0.74-0.88 (m, 2H), 1.40-1.48 (m, IH), 3.32 (s, 3H), 4.09 (s, 2H), 5.23 (s, IH), 6.69 (d, J=8.5 Hz, IH), 7.21 (dd, J=8.5, 2.3 Hz, IH), 7.38 (s, IH), 7.45 (d, J=2.3 Hz, IH), Anal. Calc'd for Cl5Hl5ClN2θ2*0.3 dioxane

C, 61.35; H, 5.53; N, 8.83 Found C, 61.05; H, 5.24; N, 8.75

EXAMPLE 5

6-chloro-4-cyclopropyl-3,4-dihydro-l-(4-methoxybenzyl)-4-(3-(tetra- hydropyran-2-vPoxy- 1 -propynyPquinazolin-2( 1 HV one

A suspension of 4.8 g (14.1 mmol) of 6-chloro-4-cyclo- propyl-l-(4-methyoxybenzyl)-quinazolin-2(lH)-one (of Example 1, Step C) and 13.6 g (42.2 mmol) of magnesium triflate was stirred for 30 minutes in 125 mL of ether under Ar at room temperature. To this solution was added a -78° solution of l-lithio-3-(tetrahydropyran-2-yl)- oxypropyne (prepared by adding 5.94 mL (42.2 mmol) of 3- (tetrahydropyran-2-yl)oxy-l-propyne dropwise to a -78° solution of 16.9 mL of 2.5 M butyllithium in hexanes and 75 mL ether) via cannula. After stirring under Ar overnight at room temperature, two additional equivalents of l-lithio-3-(tetrahydropyran-2-yl)oxypropyne (prepared as described above) were added to the reaction mixture. After 1 h, the reaction was quenched by pouring into ice-cold 1 M citric acid. The aqueous layer was extracted with EtOAc, and the combined organic layers were washed with. The organic layers were washed with 10% NaHC03, water, brine, dried over Na2S04, and the solvents removed to give an oil which was chromatographed on fine Siθ2 using 1 :2 EtOAc- hexane to afford 4.51 g (67%) of the title compound as a colorless foam: NMR (CDC13): δ 0.55-0.87 (m, 4H), 1.40-1.85 (m, 7H), 3.45-3.55 (m, IH), 3.73-3.85 (m, IH), 3.76 (s, 3H), 4.25 (s, 2H), 4.71 (t, J = 2Hz, IH), 5.11 (dd, J=34.2, 16.8 Hz, 2H), 5.28 (s, IH), 6.735 (d, J=9.0 Hz, IH), 6.835 (d, J=8.2 Hz, 2H), 7.10 (dd, J=9.0, 2.4 Hz, IH), 7.18 (d, J=8.2 Hz, 2H), 7.47 (d, J=2.4 Hz, IH). EXAMPLE 6

6-chloro-4-cyclopropyl-3,4-dihydro-3-memyl-l-(4-me oxybenzyl)-4-

(3-(tetrahydropyran-2-yl oxy-l-propynyDquinazolin-2(iHVone

A quantity of 4.1 g (8.52 mmol) of the product from Example 5 was treated by the procedure of Example 2 above to give 4.4 g of the title compound as an oil which was used in the subsequent step without further purification.

EXAMPLE 7

6-chloro-4-cyclopropyl-3,4-dihydro-3-memyl-l-(4-memoxybenzyl)-4-

(3 -hydroxy- 1 -propynyl)quinazolin-2(^' 1 HVone

The product from Example 6 (4.22 g, 0.85 mmol) was dissoved in 75 mL of ethanol and treated with 214 mg (852 mmol) of pyridinium p-toluenesulfonate at 60°C under Ar for 5 h, followed by 17 h at room temperature. The reaction mixture was concentrated and the residue partitioned between EtOAc and 10% NaHCθ3. The organic layer was washed with 10% NaHCθ3, water, brine, dried over Na2Sθ4 and solvents removed to give 3.73 g (quant.) of an off-white solid which was used in subsequent reactions without further purification.

EXAMPLE 8

6-chloro-4-cyclopropyl-3,4-dihydro-3-methyl-l-(4-methoxybenzyl)-4-

(3-chloro-l-propynyl^')quinazolin-2(^'lH)-one

To a solution of 1.25 g (3.04 mmol) of the product from Example 7 in 30 mL of CH2CI2 was added 223 mg (1.82 mmol) of 4- (dimethylamino)pyridine, 696 mg (3.65 mmol) of p-toluenesulfonyl- chloride, 424 μL (3.04 mmol) of triethylamine, and 129 mg (3.04 mmol) of lithium chloride. After stirring at room temperature under Ar overnight, the reaction mixture was diluted with 75 mL of ether and filtered. The filtrate was washed with 10% CuS04, water, 10% NaHCθ3, brine, dried over Na2S04 and solvents removed to give an oily solid which was chromatographed on fine Siθ2 using 1 :12 EtOAc - CHCI3 to afford 1.0 g (77%) of the title compound as a colorless solid: NMR (CDCI3): δ 0.40-0.57 (m, 2H), 0.68-0.86 (m, 2H), 1.31-1.38 (m, IH), 3.27 (s, 3H), 3.76 (s, 3H), 4.22 (s, 2H), 4.97 (d, J=16.2 Hz, IH), 5.21 (d, J=16.2 Hz, IH), 6.71 (d, J=8.8 Hz, IH), 6.825 (d, J=8.8 Hz, 2H), 7.11 (dd, J=8.8, 2.4 Hz, IH), 7.16 (d, J=8.8 Hz, 2H), 7.44 (d, J=2.4 Hz, IH).

EXAMPLE 9

6-chloro-4-cyclopropyl-3,4-dihydro-3-methyl-4-(3-(4-moφholinyl)-l- propynyl quinazolin-2(l HVone

A solution of 50 mg (0.116 mmol) of the product from Example 8 in 1 mL of moφholine was stirred at room temperature under Ar for 1 h, then stored in the freezer for 56 h. The solvent was removed by rotary evaporation, and the residue partitioned between water and EtOAc. The organic layer was washed with water, brine, dried over Na2Sθ4 and the solvents removed to give an oily solid which was treated by the procedure of Example 3 to afford 33 mg (78%) of the title compound as a colorless solid: mp 157-160°C;

NMR (CDCI3): δ 0.43-0.51 (m, 2H), 0.66-0.84 (m, 2H), 1.30-1.42 (m, IH), 2.59 (t, J=4.6 Hz, 4H), 3.25 (s, 3H), 3.46 (s, 2H), 3.76 (t, J=4.5 Hz, 4H), 6.76 (d, J=8.5 Hz, IH), 7.17 (dd, J=8.5, 2.2 Hz, IH), 7.379 (d, J=2.2 Hz, IH), 9.215 (s, IH).

EXAMPLE 10

6-chloro-4-cyclopropyl-3,4-dihydro-l-(4-methoxybenzyl)-4-(4-

(tetrahydropyran-2-yl)oxy-l-butyny quinazolin-2-πHVone

A quantity of 200 mg (0.587 mmol) of 6-chloro-4-cyclo- propyl-l-(4-methoxybenzyl)-quinazolin-2(lH)-one (of Example 1, Step C) was treated with l-lithio-4-((tetrahydropyran-2-yl)oxy)butyne according to the procedure of Example 5 above to afford 281 mg (97%) of the title compound as a colorless foam: NMR (CDC13): δ 0.55-0.87 (m, 4H), 1.40-1.85 (m, 7H), 3.45-3.55 (m, IH), 3.73-3.85 (m, IH), 3.76 (s, 3H), 4.25 (s, 2H), 4.71 (t, J = 2Hz, IH), 5.11 (dd, J=34.2, 16.8 Hz, 2H), 5.28 (s, IH), 6.735 (d, J=9.0 Hz, IH), 6.835 (d, J=8.2 Hz, 2H), 7.10 (dd, J=9.0, 2.4 Hz, IH), 7.18 (d, J=8.2 Hz, 2H), 7.47 (d, J=2.4 Hz, IH).

EXAMPLE 11

6-chloro-4-cyclopropyl-3,4-dihydro-l-(4-memoxybenzyl)-3-me yM-

(4-(tetrahydropyran-2-yl oxy- 1 -butynyl)quinazolin-2(l HVone

A quantity of 281 mg (0.568 mmol) of the product from Example 10 above was treated by the procedure of Example 2 above to afford 264 mg (91 %) of the title compound which was used in the subsequent reaction without further purification.

EXAMPLE 12

6-chloro-4-cyclopropyl-3,4-dihydro-l-(4-memoxybenzyl)-3-methyl-4-

(4-hydroxy-l-butynyl quinazolin-2πH)-one

A quantity of 264 mg (0.519 mmol) of the product from Example 11 was treated by the procedure of Example 7 to afford 107 mg (48%) of the title compound which was used without further purification in the subsequent reaction.

EXAMPLE 13

6-chloro-4-cyclopropyl-4-(4-fluoro-l-butynyl)-3,4-dihydro-3-methyl- quinazolin-2( 1 H -one

To a stirred solution of 107 mg (0.252 mmol) of the product from Example 12 in 2.0 mL of CH2CI2 at 0°C was added 100 μL (0.755 mmol) of diethylaminosulfurtrifluoride. The cold bath was removed, and the reaction stirred at room temperature for 2.5 h, poured into 20 mL of sat. NaHCθ3, and extracted with two portions of EtOAc. The combined organic layers were washed with water, brine, dried over MgS04 and solvents removed to give an oil which was chromatographed on 10 g of fine Siθ2 using 3:7 EtOAc-hexane to give 60 mg of a colorless oil which was treated by the procedure of Example 3 and purified by reversed phase HPLC using acetonitrile-0.1% aqueous trifluoroacetic acid on a C-18 column to give 29 mg (37 %) of the title compound as an amoφhous solid:

NMR (CDC13): δ 0.39-0.47 (m, 2H), 0.63-0.71 (m, IH), 0.76-0.84 (m, IH), 1.30-1.38 (m, IH), 2.72 (dt, J = 21.4, 6.2 Hz, 2H), 3.23 (s, 3H), 4.54 (dt, J = 46.7, 6.2 Hz, 2H), 6.76 (d, J=8.4 Hz, IH), 7.168 (dd, J=8.4, 2.4 Hz, IH), 7.362 (d, J=2.2 Hz, IH), 9.2 (s, IH).

EXAMPLE 14

6-chloro-4-(4-chloro-l-butynyl)-4-cyclopropyl-3,4-dihydro-3-methyl- quinazolin-2( 1 H V one

A quantity of 8 mg (10%) of the title compound was isolated as a byproduct from Example 13 as an amoφhous solid: NMR (CDC13): δ 0.40-0.51 (m, 2H), 0.64-0.72 (m, IH), 0.80-0.90 (m, IH), 1.30-1.38 (m, IH), 2.785 (t, J = 6.6 Hz, 2H), 3.23 (s, 3H), 3.65 (t, J= 6.6 Hz, 2H), 6.68 (d, J=8.4 Hz, IH), 7.173 (dd, J=8.4, 2.2 , z, IH), 7.42 (d, J=2.2 Hz, IH), 7.94 (s, IH).

EXAMPLE 15

6-chloro-4-cyclopropyl-3,4-dihydro-l-(4-memoxybenzyl)-4-(4-hydroxy-

1 -butynyl quinazolin-2( 1 HV one

A quantity of 75 mg (0.152 mmol) of the product from Example 10 was treated by the procedure of Example 7 to afford 61 mg (97%) of the title compound which was used without further purification in the subsequent reaction. EXAMPLE 16

6-chloro-4-cyclopropyl-4-(4-fluoro-l-butynyl)-3,4-dihydroquinazolin-

2πHVone

A quantity of 61 mg (0.148 mmol) of the product from Example 15 was treated by the procedure of Example 13 to give 7 mg (14%) of the title compound as an amoφhous solid: NMR (CDC13): δ 0.53-0.85 (m, 4H), 1.37-1.46 (m, IH), 2.60 (dt, J = 20.5, 6.5 Hz, 2H), 4.44 (dt, J = 46.6, 6.5 Hz, 2H), 5.44 (s, IH), 6.73 (d, J=8.4 Hz, IH), 7.18 (dd, J=8.4, 2.3 Hz, IH), 7.415 (d, J=2.2 Hz, IH), 8.52 (s, IH).

EXAMPLE 17

6-chloro-4-cyclopropyl-4-(3-fluoro-l-propynyl)-3,4-dihydro-3- methylquinazolin-2d H)-one

A quantity of 100 mg (0.243 mmol) of the product from Example 7 was treated by the procedure of Example 13 to give 29 mg (41 %) of the title compound as an amoφhous solid: NMR (CDC13): δ 0.46-0.57 (m, 2H), 0.68-0.84 (m, 2H), 1.34-1.42 (m, IH), 3.23 (s, 3H), 5.06 (d, J=47.2 Hz, 2H), 6.63 (d, J=8.4 Hz, IH), 7.12 (s, IH), 7.20 (dd, J=8.4, 2.2 Hz, IH), 7.397 (d, J=2.4 Hz, IH).

EXAMPLE 18

4-(3-azido-l-propynyl)-6-chloro-4-cyclopropyl-3,4-dihydro-3-methyl- quinazolin-2Cl HVone

A stirred solution of 50 mg (0.116 mmol) of the product from Example 8 was treated with 38 mg (0.582 mmol) of sodium azide in 0.5 mL dry DMF under Ar for 2 h at room temperature. The solvent was removed by rotovap, and the residue partitioned between water and EtOAc. The organic layer was washed with water, brine, dried over MgSθ4 and the solvents removed to give an oil which was treated according to the procedure of Example 3 to afford 20 mg (54%) of the title compound as an amoφhous solid:

NMR (CDC13): δ 0.48-0.58 (m, 2H), 0.69-0.77 (m, IH), 0.77-0.85 (m, IH), 1.35-1.43 (m, IH), 3.25 (s, 3H), 4.04 (s, 2H), 6.79 (d, J=8.4 Hz, IH), 7.18 (dd, J=8.4, 2.4 Hz, IH), 7.39 (d, J=2.2 Hz, IH), 9.23 (s, IH).

EXAMPLE 19

6-chloro-4-cy opropyl-3,4-dihydro-4-(3-(l-imidazolyl)-l-propynyl)-3- methylquinazolin-2(l HVone

A stirred solution of 50 mg (0.116 mmol) of the product from Example 8 was treated with 40 mg (0.582 mmol) of imidazole in 1.0 mL of DMF under Ar for 0.5 h at room temperature. The reaction was warmed to 50° for 14 hours. The solvent was removed by rotary evaporation, and the residue partitioned between water and CHCI3. The organic layer was washed with brine, dried over Na2S04 and the solvents removed to give an oil which was treated by the procedure of Example 3 to afford 15 mg (38%) of the title compound as an amoφhous solid: NMR (CDCI3): δ 0.40-0.48 (m, IH), 0.51-0.58 (m, IH), 0.67-0.75 (m, 2H), 1.34-1 42 (m, IH), 3.20 (s, 3H), 4.88 (s, 2H), 6.779 (d, J=8.4 Hz, IH), 7.03 (s, IH), 7.14 (s, IH), 7.194 (dd, J=8.5, 2.4 Hz, IH), 7.333 (d, J=2.2 Hz, IH), 7.69 (s, IH), 9.13 (s, IH).

EXAMPLE 20

6-chloro-4-cyclopropyl-3,4-dihydro-3-memyl-4-(3-(2,2,2-trifluoro- ethox v 1 -prop vn yl)quinazolin-2f 1 HVone

A solution of 20 mg (0.047 mmol) of the product from Example 8 in 1 mL of CH2CI2 was added to stirred mixture of 4.0 μL (0.047 mmol) of trifluoroethanol, 3 mg (0.051 mmol) powdered K2CO3, and 1 μL of tricaprylylmethylammonium chloride under Ar. After stirring for 3 days at room temperature, the reaction was diluted with 15 mL of CHCI3 and washed with water, brine, dried over Na2Sθ4 and the solvents removed to give an oil which was chromatographed on 3 g fine Siθ2 using 1 :3 EtOAc-hexane to give an oil which was treated by the procedure of Example 3 to afford 6 mg (13%) of the title compound as a solid: mp 128-129°C;

NMR (CDC13): δ 0.42-0.51 (m, IH), 0.51-0.57 (m, IH), 0.68-0.80 (m, 2H), 1.34-1.42 (m, IH), 3.22 (s, 3H), 3.90 (q, J = 8.6 Hz, 2H), 4.42 (s, 2H), 6.645 (d, J=8.4 Hz, IH), 7.191 (dd, J=8.4, 2.2 Hz, IH), 7.23 (s, IH), 7.385 (d, J=2.2 Hz, IH).

EXAMPLE 21

6-chloro-4-cyclopropyl-3,4-dihydro-3-methyl-4-(3-(4-pyridyloxy)-l- propynyl)quinazolin-2(l HVone

A stirred solution of 50 mg (0.116 mmol) of the product from Example 8 was treated with 96 mg (0.349 mmol) of silver carbonate and 12 mg (0.128 mmol) of 4-hydroxypyridine in 2.0 mL of DMF under Ar for 1.5 h at room temperature. The reaction was warmed to 50° overnight. The solvent was removed by rotary evaporation, and the residue partitioned between water and EtOAc. The organic layer was washed with water, brine, dried over Na2Sθ4 and the solvents removed to give an oil which was chromatographed on fine Siθ2 using 1 :3 CHCl3-EtOAc to afford 20 mg of an oil which was treated by the procedure of Example 3 to afford 15 mg (17%) of the title compound as a solid: mp 180-184°C;

NMR (CDC13): δ 0.35-0.47 (m, IH), 0.48-0.58 (m, IH), 0.63-0.72 (m, 2H), 1.28-1.38 (m, IH), 3.16 (s, 3H), 4.90 (s, 2H), 6.77 (d, J=8.4 Hz, IH), 6.98 (d, J=6.5 Hz, 2H), 7.18 (dd, J=8.4, 2.3 Hz, IH), 7.29 (d, J=2.3 Hz, IH), 8.55 (br d, J=6 Hz, 2H), 8.78 (s, IH). EXAMPLE 22

6-chloro-4-cyclopropyl-3,4-dihydro-3-methyl-4-(3-(4-(N- oxopyridyPoxy )- 1 -propynvDquinazolin-2( 1 H)-one

The title compound (41 mg, 46%) was isolated as a byproduct from Example 21 as a solid: mp 139-141 °C; NMR (CDCI3+CD3OD): δ 0.42-0.53 (m, IH), 0.60-0.82 (m, 3H), 1.34- 1.45 (m, IH), 3.16 (s, 3H), 5.07 (s, 2H), 6.75 (d, J=8.5 Hz, IH), 6.90-6.97 (m, 2H), 7.21 (dd, J=8.5, 1.8 Hz, IH), 7.33 (d, J=2.2 Hz, IH), 8.00 (d, J=7.4 Hz, 2H).

EXAMPLE 23

6-chloro-4-cyclopropyl-3,4-dihydro-l-(4-methoxybenzyl)-4-((2- pyridyl)ethynyDquinazolin-2f 1 HVone

A suspension of 200 mg (0.59 mmol) of 6-chloro-4-cyclo- propyl-l-(4-methoxybenzyl)-quinazolin-2(lH)-one (of Example 1, Step C) and 567 mg (1.76 mmol) of magnesium triflate was stirred for 30 minutes in 10 mL of ether under Ar at room temperature. In a separate flask, 181 mg (1.76 mmol) of 2-ethynylpyridine was dissolved in 10 mL of dry THF under Ar, cooled to -78°C, and treated with 704 μL of 2.5 M n-butyllithium in hexanes. After this solution was stirred at -78°C for 0.5 h, it was added dropwise to the ether suspension described above. The cold bath was removed and stirring c ntinued at room temp, for 2.5 h. A second 1.76 mmol portion of l-lithio-2-(2-pyridyl)acetylene solution was added to the reaction mixture which was stirred overnight to complete the reaction. The reaction was quenched by pouring into 10% citric acid and extracted with two portions of EtOAc. The organic layers were washed with water, brine, dried over MgSθ4 and solvents removed to give an oil which was chromatographed on fine Siθ2 using 1 :3 hexanes-EtOAc to provide 183 mg (70%) of the title compound as a yellow solid which was used without further purification in the subsequent steps. EXAMPLE 24

6-chloro-4-cyclopropyl-3,4-dihydro-4-((2-pyridyl)ethynyl)quinazolin-

2(TH)-one

A quantity of 70 mg (0.16 mmol) of the product from Example 23 was treated by the procedure of Example 3 for 96 h to afford 38 mg (73%) of the title compound as an amoφhous solid: NMR (CDC13): δ 0.58-0.72 (m, IH), 0.73-0.90 (m, 2H), 0.91-1.04 (m, IH), 1.47-1.60 (m, IH), 5.85 (s, IH), 6.78 (d, J=8 Hz, IH), 7.15 (dd, J=8, 2 Hz, IH), 7.20-7.28 (m, IH), 7.39 (d, J=8 Hz, IH), 7.52 (d, J=2 Hz, IH), 7.63 (td, J=8, 2 Hz, IH), 8.58(d, J=4 Hz, IH), 9.13 (s, IH).

EXAMPLE 25

6-chloro-4-cyclopropyl-3,4-dihydro-4-((3-pyridyl)ethynyl)quinazolin-

2αHVone

A quantity of 300 mg (0.88 mmol) of 6-chloro-4-cyclo- propyl-l-(4-methyoxybenzyl)-quinazolin-2(lH)-one (of Example 1, Step C) was treated with 3-ethynylpyridine (prepared according to Sakamoto et al, Synthesis, No. 1, p. 312, 1983) by the procedure of Example 23 to afford 187 mg of a yellow solid. A quantity of 100 mg of this material was treated by the procedure of Example 3 to provide 68 mg (39%) of the title compound as a colorless foam. An analytical sample was obtained by trituration with ether-hexane: mp 231-233 °C; NMR (CDC13): δ 0.6-0.78 (m, 2H), 0.79-0.95 (m, 2H), 1.49-1.60 (m, IH), 5.90-6.20 (m, IH), 6.84 (d, J=8 Hz, IH), 7.22 (dd, J=8, 2 Hz, IH), 7.32-7.41 (m, IH), 7.49 (s, IH), 7.76-7.84 (m, IH), 8.40 (br s, IH), 8.50- 8.62 (m, IH), 8.75-8.90 (m, IH). EXAMPLE 26

6-chloro-4-cyclopropyl-3,4-dihydro-4-((4-pyridyl)ethynyl)quinazolin-

2QH -one

A quantity of 250 mg (0.73 mmol) of 6-chloro-4-cyclo- propyl-l-(4-methyoxybenzyl)-quinazolin-2(lH)-one (of Example 1, Step C) was treated with 4-ethynylpyridine (prepared according to Sakamoto et al, supra) by the procedure of Example 23 to afford 155 mg of a colorless crystalline solid, mp 157-160°C. A quantity of 125 mg of this material was treated by the procedure of Example 3 to provide 58 mg (32%) of the title compound as a solid: mp 131-133°C; NMR (CDC13): δ 0.63-0.76 (m, IH), 0.77-0.81 (m, IH), 0.84-0.96 (m, IH), 0.97-1.04 (m, IH), 1.58-1.63 (m, IH), 5.62 (s, IH), 6.83 (d, J=8 Hz, IH), 7.20-7.40 (m, IH), 7.51 (s, IH), 7.76 (s, IH), 7.80-7.83 (m, 2H), 8.74-8.76 (m, 2H).

EXAMPLE 27

6-chloro-4-cyclopropyl-3,4-dihydro-4-((2-pyrazinyl)ethynyl)quinazolin-

2(lH -one

A quantity of 120 mg (0.352 mmol) of 6-chloro-4-cyclo- propyl-l-(4-methyoxybenzyl)-quinazolin-2(lH)-one (of Example 1, Step C) was treated with 2-ethynylpyrazine (prepared according to Sakamoto et al, supra) by the procedure of Example 23 as in Step W to afford 110 mg of an oil which was then treated by the procedure of Example 3 to provide 33 mg (29%) of the title compound as a solid: mp 245°C (dec); NMR (DMSO-d6): δ 0.50-0.60 (m, IH), 0.61-0.72 (m, 2H), 0.76-0.84 (m, IH), 1.44-1.52 (m, IH), 6.88 (d, J=8.6 Hz, IH), 7.30 (dd, J=8.6, 2.4 Hz, IH), 7.467 (d, J=2.4 Hz, IH), 7.78 (d, J=1.7 Hz, IH), 8.62-8.66 (m, 2H), 8.737 (d, J=1.3 Hz, IH), 9.64 (s, IH). EXAMPLE 28

6-chloro-4-cyclopropyl-3,4-dihydro-4-((5-pyrimidinyl)ethynyl)- quinazolin-2(l H)-one

A quantity of 300 mg ( 0.88 mmol) of 6-chloro-4-cyclo- propyl-l-(4-methyoxybenzyl)-quinazolin-2(lH)-one (of Example 1, Step C) was treated with 5-ethynylpyrimidine (prepared according to Sakamoto et al, supra) according to the procedure of Example 23 to afford 125 mg of a yellow solid, mp 165-167°C, which was then treated by the procedure of Example 3 to provide 49 mg (18%) of the title compound as a solid: mp 255-256°C (dec);

NMR (CDCl3-DMSO-d6): δ 0.60-0.92 (m, 4H), 1.47-1.57 (m, IH), 6.54 (s, IH), 6.91 (d, J=8.4 Hz, IH), 7.177 (dd, J=8.6, 2.4 Hz, IH), 7.456 (d, J=2.2 Hz, IH), 7.622 (s, IH), 8.70-8.90 (m, 2H), 9.10-9.20 (m, IH), 9.45 (s, IH).

EXAMPLE 29

6-chloro-4-cyclopropyl-3,4-dihydro-3-methyl-4-((2-pyridyl)ethynyl)- quinazolin-2(l HVone

A quantity of 95 mg (0.21 mmol) of the product from Example 23 was treated by the procedure of Example 2 to afford 78 mg of an oil which was then treated according to the procedure of Example 3 to afford 39 mg (54%) of the title compound as a colorless solid: mp 185.5-186.5°C;

NMR (CDC13): δ 0.53-0.65 (m, 2H), 0.73-0.80 (m, IH), 0.91-0.98 (m, IH), 1.44-1.51 (m, IH), 3.33 (s, 3H), 6.73 (d, J=8.4 Hz, IH), 7.20 (dd, J=8.4, 2.2 Hz, IH), 7.28-7.32 (m, IH), 7.47 (d, J=8 Hz, IH), 7.51 (d, J=2.2 Hz, IH), 7.697 (td, J=7.7, 1.6 Hz, IH), 8.3 (s, IH), 8.62-8.64 (m, IH). EXAMPLE 30

6-chloro-4-cyclopropyl-3,4-dihydro-l-(4-methoxvbenzyl)-4-ethynyl)- quinazolin-2f 1 H V one

A quantity of 5.0 g (14.67 mmol) of 6-chloro-4-cyclo- propyl-l-(4-methyoxybenzyl)-quinazolin-2(lH)-one (of Example 1, Step C) was treated with (trimethylsilyl)acetylene according to the procedure of Example 23 to afford approximately 7 g of an oil which was dissolved in 200 mL of THF and stirred vigorously with 150 mL of IM KOH for 20 minutes at room temperature. The reaction was acidified with 3M HCl and extracted with two portions of ether. The organic layers were combined and washed with water, brine, dried over MgSθ4 and the solvents removed to give an oily yellow solid which was triturated w'th ether-hexanes, followed by trituration with acetonitrile to afford the title compound as a colorless solid. All of the trituration filtrates were combined, concentrated and retriturated with acetonitrile to give a colorless solid which provided a combined yield of 3.54 g (66%) of the title compound:

NMR (CDCI3): δ 0.59-0.72 (m, 2H), 0.77-0.90 (m, 2H), 2.52 (s, IH), 3.77 (s, 3H), 5.04 (d, J=16.3 Hz, IH), 5.17 (d, J=16.3 Hz), 5.34 (s, IH), 6.755 (d, J=8.8 Hz, IH), 6.845 (d, J=8.8Hz, 2H), 7.125(dd, J=8.8, 2.4 Hz, IH), 7.193 (d, J=8.79, 2H), 7.50 (d, J=2.4 Hz, IH).

EXAMPLE 31

6-Chloro-4-cyclopropyl-4-ethynyl-3.4-dihydroquinazolin-2(lHVone

A solution of 1.4 g (4.11 mmol) of the product from Example 30 in 5 mL of CH2CI2 was treated with 10 mL of trifluoroacetic acid under N2 overnight at room temperature. The reaction was concentrated by rotary evaporation under reduced pressure and the residue partitioned between ethyl acetate and 10% citric acid. The organic layer was washed with water, brine, dried over MgSθ4 and the solvents removed to give an oil which was flash chromatographed on Siθ2 using 95:5 CHCI3-CH3OH to give a foam. Trituration of this material with ether gave 720 mg (80%) of the title compound as a colorless solid.

EXAMPLE 32

6-chloro-4-cyclopropyl-3,4-dihydro-4-((2-pyrimidmyl)ethynyl)- quinazolin-2( 1 H V one

A mixture of 87 mg (0.35 mmol) of 6-chloro-4-cyclo- propyl-3,4-dihydro-4-ethynylquinazolin-2(lH)-one (the product of Example 31), 111 mg (0.7 mmol) of 2-bromopyrimidine, 13 mg (0.018 mmol) of bis(triphenylphosphine) palladium di chloride, and 1.5 mL of triethylamine was stirred in a sealed tube at 80°C overnight. After cooling, the reaction was diluted with methanol, filtered through a Celite pad, and concentrated to give an oily solid which was chromatographed on fine Siθ2 using 95:5 CHCI3-CH3OH 0 provide 75 mg (66%) of the title compound as a colorless solid. An analytical sample was obtained by crystallization from ether-chloroform: mp 259-261°C (dec); NMR (CDCl3-DMSO-d6): δ 0.61-0.73 (m, IH), 0.77-0.87 (m, 2H), 0.95-1.03 (m, IH), 1.51-1.59 (m, IH), 6.03 (s, IH), 6.88 (d, J=8.4 Hz, IH), 7.169 (dd, J=8.6, 2.4 Hz, IH), 7.31 (t, J=4.8 Hz, IH), 7.52 (d, J = 2.2 Hz, IH), 8.726 (d, J=4.7 Hz, 2H), 9.27 (s, IH).

EXAMPLE 33

6-chloro-4-cyclopropyl-3,4-dihydro-3-memyl-4-(3-(N,N-dimemyl^ aminoV 1 -propynyl quinazolin-2( 1 HI -one

A 75 mg (0.175 mmol) sample of the product from Example 8 was treated with 5 mL of dimethylamine (condensed at -78°C) in a pressure tube. The stirred solution was allowed to warm to room temperature over a 4 h period. After the dimethylamine was allowed to evaporate, the residue was partitioned between CHCI3 and 10% NaHC03. The organic layer was washed with water, brine, dried over Na2S04 and concentrated to give 79 mg of an oil which was treated according to Example 3 to afford 52 mg (93%) of the title compound as a colorless solid: mp 135-137°C;

NMR (CDC13): δ 0.42-0.54 (m, 2H), 0.63-0.77 (m, IH), 0.78-0.90 (m, IH), 1.30-1.42 (m, IH), 2.36 (s, 6H), 3.25 (s, 3H), 3.45 (s, 2H), 6.78 (d, 3=8.4 Hz, IH), 7.17 (dd, J=8.4, 2.3 Hz, IH), 7.395 (d, J=2.2 Hz, IH), 9.30 (s, IH).

EXAMPLE 34

6-Chloro-4-cycloρropyl-3,4-dihydro-4-(phenylethynyl)quinazolin-2(lH)- one

A mixture of 70 mg (0.28 mmol) of 6-Chloro-4-cyclo- propyl-3,4-dihydro-4-ethynylquinazolin-2(lH)-one (Example 31), was coupled with iodobenzene according to the methods of Example 32 to provide 50 mg of the title compound as a colorless solid: mp 193- 195°C(dec);

NMR (CDC13): δ 0.59-0.67 (m, IH), 0.72-0.85 (m, 2H), 0.86-0.98 (m, IH), 1.48-1.56 (m, IH), 5.44 (s, IH), 6.74 (d, J=8.4 Hz, IH), 7.21 (dd, J=8.6, 2.3 Hz, IH), 7.25-7.40 (m, 5H), 7.51 (d, J=2.3 Hz, IH), 8.05 (s,lH).

EXAMPLE 35

4-(3-buten-l-ynyl)-6-chloro-4-cyclopropyl-3,4-dihydro-3-methyl- quinazolin-2πHVone

A solution of 200 mg (0.484 mmol) of the protected, penultimate product of Example 13 (before TFA treatment according to Example 3) was methylated according to Example 2 to give 200 mg of an oil. Treatment of this oil with TFA according to Example 3 provided 20 mg of the title compound as a colorless solid: mp 123-124°C, NMR (CDCI3): δ 0.40-0.53 (m, 2H), 0.63-0.73 (m, IH), 0.73-0.85 (m, IH), 1.32-1.43 (m, IH), 3.24 (s, 3H), 5.56-5.93 (m, 3H), 6.77 (d, J=8.3 Hz, IH), 7.17 (dd, J=8.4, 1.9 Hz, IH), 7.37 (d, J=1.9 Hz, IH), 9.19 (s, IH). EXAMPLE 36

6-Chloro-4-cyclopropyl-3,4-dihydro-4-(3-hydroxy-l-propynyl)-3- methy lquinazolin-2( 1 HVone

A sample of 344 mg (0.837 mmol) of the product from Example 7 was deprotected by treatment with TFA according to Example 3, to give 200 mg (82%) of the title compound as a colorless solid. An analytical sample was obtained by crystallization from hexane: mp 76- 80°C;

NMR (CDC13): δ 0.43-0.53 (m, 2H), 0.65-0.75 (m, IH), 0.75-0.84 (m, IH), 1.32-1.39 (m, IH), 3.22 (s, 3H), 4.39 (s, 2H), 6.68 (d, J=8.4 Hz, IH), 7.17 (dd, J=8.4, 2.4 Hz, IH), 7.39 (d, J=2.2 Hz, IH), 7.90 (s, IH).

EXAMPLE 37

6-Chloro-4-cyclopropyl-3,4-dihydro-3-memyl-4-(3-(2-pyridyloxy)-l- propynyl)quinazolin-2(l HVone

A sample of 63 mg (0.129 mmol) of the product from Example 8 and 99.7 mg (1.05 mmol) of 2-hydroxypyridine was treated according to the procedure of Example 21 to give 34 mg (71 %) of the title compound as a colorless solid: mp 130-132°C; NMR (CDCI3): δ 0.30-0.41 (m, 2H), 0.54-0.65 (m, IH), 0.65-0.76 (m, IH), 1.28-1.38 (m, IH), 3.18 (s, 3H), 5.05 (s, 2H), 6.72 (d, J=8.4 Hz, IH), 6.81 (d, J=8.4 Hz, IH), 6.92-6.96 (m, IH), 7.15 (dd, J=8.4 2.3 Hz, IH), 7.33 (d, J=2.2 Hz, IH), 7.59-7.65 (m, IH), 8.22 (dd, J=5.1, 2.0 Hz, IH), 8.96 (s, IH). EXAMPLE 38

6-Chloro-4-cyclopropyl-3,4-dihydro-4-(2-nitrophenylethynyl)- quinazolin-2( 1 H V one

A mixture of 60 mg (0.24 mmol) of 6-chloro-4-cyclo- propyl-3,4-dihydro-4-ethynylquinazolin-2(lH)-one (Example 31), was coupled with 2-iodonitrobenzene according to the procedure of Example 32 to provide 32 mg of the title compound as a colorless solid: mp 181- 182°C(dec);

NMR (CDC13): δ 0.63-0.71 (m, IH), 0.77-0.91 (m, 2H), 1.00-1.85 (m, IH), 1.50-1.58 (m, IH), 5.51 (s, IH), 6.76 (d, J=8.6 Hz, IH), 7.23 (dd, J=8.4 2.4 Hz, IH), 7.45-7.55 (m, IH), 7.55-7.60 (m, 3H), 8.01 (s, IH), 8.08 (d, J=8.05 Hz, IH).

EXAMPLE 39

l,3-(di-(lS)-camphanoyl)-6-chloro-4-cyclopropyl-3,4-dihydro-4-((2- pyridyl ethynyl quinazolin-2( 1 H V one

A solution of 200 mg (0.618 mmol) of the product from Example 24, 134 mg (0.618 mmol) of (lS)-camphanic chloride, 76 mg (0.618 mmol) of N,N-dimethylaminopyridine (DMAP), and 0.43 mL (3.09 mmol) of triethylamine in 2.0 mL of CH2CI2 was stirred under Ar at room temperature for 18 hours. An additional 76 mg (0.618 mmol) of DMAP and 268 mg (1.23 mmol) of (lS)-camphanic chloride was added to the reaction mixture, and stirring continued for 6 hours. The reaction was diluted with CHCI3 and washed with IM citric acid, water, 10% Na2Cθ3, dried over Na2Sθ4 and treated with activated carbon. Removal of the solvents gave a yellow foam which was chromatographed on 50 g fine Siθ2 using 1 :2 EtOAc-hexane. The early eluting fractions were combined and evaporated to give 174 mg of diasteromer 1 as an almost colorless foam. Diasteromer 2 was obtained upon further elution as 138 mg of a foam. An analytical sample of diasteromer 2 was obtained by trituration from methanol; Calc'd for C38H38CIN3O7

C, 66.71; H, 5.60; N, 6.14 Found C, 66.38; H, 5.53; N, 6.17

EXAMPLE 40

(-)6-chloro-4-cyclopropyl-3,4-dihydro-4-((2-pyridyl)ethynyl)-quinazolin-

2(lHVone

A solution of 143 mg (0.254 mmol) of diasteromer 1 from Example 39 in 1.0 mL of dimethoxyethane was treated with 0.4 mL of 1.0 M aq. LiOH under Ar for 1.5 h. The reaction was partitioned between EtOAc and water. The aqueous layer was extracted with EtOAc and the combined organic layers were washed with water, brine, dried over MgS04, and the solvents removed to give 115 mg of a foam. This material was dissolved in 2.0 mL of ethanol, treated with 32 mg (0.168 mmol) of p-toluene sulfonic acid and heated at reflux under Ar for 64 h. The solvents were removed by rotary evaporation at reduced pressure and the residue partitioned between 10% Na2Cθ3 and EtOAc. The aqueous layer was extracted with EtOAc, and the combined organic layers washed with water, brine, dried over MgS04, treated with activated carbon, and the solvents removed to give a solid which was triturated with 1 :1 Et2θ- hexane to provide 16 mg of the title compound as a pale yellow solid: NMR (CDC13) same as for Example 24; 0©= -100° (c = 0.4, CHCI3).

REVERSETRANSCRIPTASEASSAY

The assay measures the incoφoration of tritiated deoxyguanosine monophosphate by recombinant HTV reverse transcriptase (HIV RTR) (or other RT) into acid-precipitable cDNA at the Km values of dGTP and poly r(C oligo d(G)l2-18- The inhibitors of the present invention inhibit this incoφoration.

The assays were carried out in 55 mM Tris (pH 8.2)-30 mM KC1-30 mM MgCl2-l mM dithiothreitol-20 μg of rC:dGl2-18 (Pharmacia) per ml-8 μM [ H]dGTP (New England Nuclear)-0.01% Triton X-l 00-50 μM ethylene glycol-bis(β-amino-ethyl ether)-N,N,N',N^*- tetraacetic acid (EGTA)-l mg of bovine serum albumin per ml. After 60 minutes of incubation at 37°C, acid-precipitable material was collected onto glass fiber filters by using a semiautomatic cell harvester. Bacterial cell extracts containing RT were diluted to within the linear range of the assay, and activity was determined in the presence and absence of inhibitor. Purified HIV-1 RT heterodimer produced in E. coli also served as a control.

For the double (DBL) mutant assay, A17 RT was employed in the assay. A17 RT is resistant to various aminopyridones, as described in Nunberg, J.H. et al, J. Virol, 65, 4887 (1991).

INHIBITION OF VIRUS SPREAD

A. Preparation of HTV-infected MT-4 cell Suspension. MT cells were infected at Day 0 at a concentration of

250,000 per ml with a 1 : 1000 dilution of HIV-1 strain IHb stock (final 125 pg p24/ml; sufficient to yield <1% infected cells on day 1 and 25- 100% on day 4). Cells were infected and grown in the following medium: RPMI 1640 (Whittaker BioProducts), 10% inactivated fetal bovine serum, 4 mM glutamine (Gibco Labs) and 1 :100 Penicillin- Streptomycin (Gibco Labs).

The mixture was incubated overnight at 37°C in 5% Cθ2 atmosphere.

B. Treatment with Inhibitors

A matrix of nanomolar range concentrations of the pairwise combinations (see Table S) was prepared. At Day 1, aliquots of 125 μl of inhibitors were added to equal volumes of HlV-infected MT-4 cells (50,000 per well) in a 96-well microtiter cell culture plate. Incubation was continued for 3 days at 37°C in 5% Cθ2 atmosphere. C. Measurement of Virus Spread

Using a multichannel pipettor, the settled cells were resuspended and 125 μl harvested into a separate microtiter plate. The supernatant was assayed for HIV p24 antigen.

The concentration of HIV p24 antigen was measured by an enzyme immunoassay, described as follows. Aliquots of p24 antigen to be measured were added to microwells coated with a monoclonal antibody specific for HIV core antigen. The microwells were washed at this point, and at other appropriate steps that follow. Biotinylated HIV- specific antibody was then added, followed by conjugated strepavidin- horseradish peroxidase. A color reaction occurs from the added hydrogen peroxide and tetramethylbenzidine substrate. Color intensity is proportional to the concentration of HIV p24 antigen.

Calculation of Degree of Synergy

Pairwise combinations of inhibitors (see Table S) were found to exhibit markedly enhanced inhibition of virus spread, in comparison to each inhibitor alone, or in comparison to merely additive inhibition of each inhibitor. Thus, for example, the pairwise combination of 372 and ddl was found to exhibit markedly enhanced inhibition of virus spread, in comparison to 372 alone or ddl, or in comparison to the sum of 372 inhibitor and ddl inhibition.

This data was processed as follows: fractional inhibitory concentration ratios (F1C) were calculated according to Elion, et al, J. Biol. Chem., 208, 477 (1954). The minimum sum of FICS, which is the maximum synergy, was determined for various pairwise combinations. See Table S. These results indicate substantial synergy in the inhibition of virus spread. The smaller the number, the greater the synergy. TAB^TJE S

Pairwise Combinations* Maximum Svnergv

372 + ddl 0.3-0.4 372 + AZT 0.6-0.8 372 + 524 0.7

*372 is 6-chloro-4(S)-cyclopropyl-3,4-dihydro-4-((2-pyridyl)- ethynyl)quinazolin-2(lH)-one, which is compound 26. 524 is L-735,524 (Table C). Other compounds are also defined in Table C above.

While the foregoing specification teaches the principles of the present invention, with examples provided for the puφose of illustration, it will be understood that the practice of the invention encompasses all of the usual variations, adaptations, or modifications, as come within the scope of the following claims and its equivalents.

Claims

WHAT IS CLAIMED IS:

1. A compound of the formula:

I wherein:

X is O,

G, when present, is halo, nitro, or cyano; n is 0-4;

Rl is C3-5cycloalkyl, C2-5 alkynyl, C2-4alkenyl, or cyano;

R2 is C2-5alkynyl substituted with one or more of A, or C2-5 alkenyl substituted with one or more of A, wherein A is i) halo; ii) hydroxy; iii) amino; iv) cyano; v) nitro; vi) azido; vii) C3-8 cycloalkyl; viii) Cl -4 alkoxy, unsubstituted or substituted with one or more of halo; ix) di-(Cl-4alkyl) amino; x) Cl-4 alkylamino; xi) aryl, unsubstituted or substituted with one or more of D, wherein D is amino, nitro, cyano, or Cl-3 alkoxy; xii) aryloxy, unsubstituted or substituted with one or more of D; xiii) heterocycle, unsubstituted or substituted with one or more of D; xiv) heterocycle-oxy; or xv) C2-5 alkenyl; xvi) COOR, wherein R is H, C l -4alkyl or aryl; xvii) CONR2; or xviii) COR;

R3 is i) H; ii) cyano; iii) amino; iv) hydroxyl; v) Cl-4 alkyl, unsubstituted or substituted with one or more of

E, wherein E is halo, hydroxyl, amino, nitro, cyano,

Cl-4- alkoxy, or C3-5 cycloalkyl; vi) C2-4 alkenyl, unsubstituted or substituted with E; or vii) C2-4 alkynyl, unsubstituted or substituted with E;

i) H; ii) Cl-4 alkyl; iii) C l -5 alkylcarbonyl; iv) benzoyl, unsubstituted or substituted with one or more of A; or v) heterocyclecarbonyl; with the proviso that any terminal alkynyl carbon is not substituted with any substituent selected from the group consisting of halo, hydroxy, amino, cyano, nitro, azido, Ci-4alkoxy unsubstituted or substituted with one or more of halo, di-(Cl-4alkyl)amino, Cl-4alkylamino, aryloxy unsubstituted or substituted with one or more of D, or heterocycleoxy; or pharmaceutically acceptable salt thereof. A compound according to Claim 1 , of the formula

π wherein:

R2 is C2-5 alkynyl substituted with halo, hydroxy, amino, cyano, nitro, azido, C3-8 cycloalkyl, Cl-4 alkoxy, di-(Cι_4-alkyl)- amino, Cl-4 alkylamino, phenyl,

2-nitrophenyl, pyridyl, pyrimidyl, pyrazinyl, imidazolyl, or C2-3 alkenyl;

R3 is H or C 1-3 alkyl; or pharmaceutically acceptable salt thereof.

3. A compound which is 6-chloro-4-cyclopropyl-3,4-dihydro-3-methyl-4-(3-methoxy-l - propynyl)quinazolin-2(l H)-one,

6-chloro-4-cyclopropyl-3,4-dihydro-4-(3-methoxy-l-propynyl)- quinazolin-2(lH)-one,

6-chloro-4-cyclopropyl-3,4-dihydro-3-methyl-4-(3-(4-moφholinyl)-l- propynyl)quinazolin-2(lH)-one,

6-chloro-4-cyclopropyl-4-(4-fluoro-l-butynyl)-3,4-dihydro-3-methyl- quinazolin-2(lH)-one,

6-chloro-4-(4-chloro-l-butynyl)-4-cyclopropyl-3,4-dihydro-3-methyl- quinazolin-2(l H)-one, 6-chloro-4-cyclopropyl-4-(4-fluoro-l -butynyl)-3,4-dihydroquinazolin-

2(lH)-one,

6-chloro-4-cyclopropyl-4-(3-fluoro-l-propynyl)-3,4-dihydro-3-methyl- quinazolin-2(l H)-one,

4-(3-azido-l-propynyl)-6-chloro-4-cyclopropyl-3,4-dihydro-3-methyl- quinazolin-2(l H)-one, 6-chloro-4-cyclopropyl-3,4-dihydro-4-(3-(l-imidazolyl)-l-propynyl)-3- methylquinazolin-2(l H)-one,

6-chloro-4-cyclopropyl-3,4-dihydro-3-methyl-4-(3-(2,2,2,-trifluoro- ethoxy)- 1 -propyny l)quinazolin-2( 1 H)-one,

6-chloro-4-cyclopropyl-3,4-dihydro-3-methyl-4-(3-(4-pyridyloxy)-l- propynyl)quinazolin-2(lH)-one,

6-chloro-4-cyclopropyl-3,4-dihydro-4-((2-pyridyl)ethynyl)quinazolin-

2(lH)-one,

6-chloro-4-cyclopropyl-3,4-dihydro-4-((3-pyridyl)ethynyl)quinazolin-

2(lH)-one,

6-chloro-4-cyclopropyl-3,4-dihydro-4-((4-pyridyl)ethynyl)quinazolin-

2(lH)-one,

6-chloro-4-cyclopropyl-3,4-dihydro-4-((2-pyrazinyl)ethynyl)quinazolin-

2(lH)-one,

6-chloro-4-cyclopropyl-3,4-dihydro-4-((5-pyrimidinyl)ethynyl)- quinazolin-2(lH)-one,

6-chloro-4-cyclopropyl-3,4-dihydro-3-methyl-4-((2-pyridyl)ethynyl)- quinazolin-2(l H)-one,

6-chloro-4-cyclopropyl-3,4-dihydro-4-((2-pyrimidinyl)ethynyl)- quinazolin-2( 1 H)-one ,

6-chloro-4-cyclopropyl-3,4-dihydro-3-methyl-4-(3-(N,N-dimethyl- amino)- 1 -propynyl)quinazolin-2(lH)-one,

6-chloro-4-cyclopropyl-3,4-dihydro-4-(phenylethynyl)quinazolin-2(lH)- one,

4-(3-buten- 1 -ynyl)-6-chloro-4-cyclopropyl-3,4-dihydro-3-methyl- quinazolin-2( 1 H)-one,

6-chloro-4-cyclopropyl-3,4-dihydro-4-(3-hydroxy-l-propynyl)-3- methylquinazolin-2( 1 H)-one,

6-chloro-4-cyclopropyl-3,4-dihydro-3-methyl-4-(3-(2-pyridyloxy)-l- propynyl)quinazolin-2(l H)-one,

6-chloro-4-cyclopropyl-3,4-dihydro-4-((2-nitrophenyl)ethynyl)- quinazolin-2(lH)-one, or

6-chloro-4(S)-cyclopropyl-3,4-dihydro-4-((2-pyridyl)ethynyl)- quinazolin-2(lH)-one, or pharmaceutically acceptable salt thereof.

4. A compound of Claim 3, which is 6-chloro-4-cyclopropyl-4-(4-fluoro-l-butynyl)-3,4-dihydro-3-methyl- quinazolin-2(lH)-one,

6-chloro-4-cyclopropyl-3,4-dihydro-3-methyl-4-((2-pyridyl)ethynyl)- quinazolin-2( 1 H)-one ,

6-chloro-4-cyclopropyl-3,4-dihydro-4-((2-pyridyl)ethynyl)quinazolin- 2(lH)-one,

6-chloro-4-cyclopropyl-3,4-dihydro-4-(phenylethynyl)quinazoin-2(lH)- one, or

6-chloro-4(S)-cyclopropyl-3,4-dihydro-4-((2-pyridyl)ethynyl)quina- zolin-2(lH)-one, or pharmaceutically acceptable salt thereof.

5. The synergistic combination of 6-chloro-4(S)-cyclopropyl-3,4-dihydro-4-((2-pyridyl)ethynyl)- quinazolin-2(lH)-one, and ddl.

6. The synergistic combination of 6-chloro-4(S)-cyclopropyl-3,4-dihydro-4-((2-pyridyl)ethynyl)- quinazolin-2(lH)-one, and AZT.

7. The synergistic combination of 6-chloro-4(S)-cyclopropyl-3,4-dihydro-4-((2-pyridyl)ethynyl)- quinazolin-2(lH)-one, and

N-(2(R)-hydroxy- 1 (S)-indanyl)-2(R)-phenylmethyl-4-(S)-hydroxy-5- ( 1 -(4-(3 -pyridylmethyl)-2(S)-N'-(t-butylcarboxamido)-piperazinyl))- pentanemamide.

8. A method of inhibiting HIV reverse transcriptase, comprising administering to a mammal an effective amount of a compound as in any of Claims 1-4 or an effective amount of a synergistic combination as in any of Claims 5-7.

9. A method of preventing infection of HIV, or of treating infection by HIV or of treating AIDS or ARC, comprising administering to a mammal a pharmaceutically acceptable carrier with an effective amount of a compound as in any of Claims 1-4, or an effective amount of a synergistic combination as in any of Claims 5-7.

10. A pharmaceutical composition useful for inhibiting HIV reverse transcriptase, comprising a pharmaceutically acceptable carrier and an effective amount of a compound as in any of Claims 1-4, or an effective amount of a synergistic combination as in any of Claims

5-7.

11. A pharmaceutical composition useful for preventing or treating infection of HIV or for treating AIDS or ARC, comprising a pharmaceutically acceptable carrier and an effective amount of a compound as in any of Claims 1-4, or an effective amount of a synergistic combination as in any of Claims 5-7.

12. The compound 6-chloro-4(S)-cyclopropyl-3,4- dihydro-4-((2-pyridyl)ethynyl)quinazolin-2( 1 H)-one monohydrate.