LV12719B - Benzoksazinons kā hiv apgrieztās transkriptāzes inhibitors - Google Patents

Description

Description [0001] A retrovirus designated human immunodeficiency virus (HIV) is the etiological aģent of the complex disease that includes Progressive destruction ofthe immune system (acquired immune deficiency syndrome; AIDS) and degeneration ofthe Central and peripheral nervous system. This virus vvas previously knovvn as LAV, HTLV-ilI, or ARV. A common feature of retrovirus replication is reverse transcription of the RNA genome by a virally encoded reverse transcriptase to generate DNA copies of HIV sequences, a required step in virai replication. It is knovvn that some compounds are reverse transcriptase inhibitors and are effective aģents in the treatment of AIDS and similar cfiseases, e.g. azidothymidine or AZT.

[0002] Nudeotide sequencing of HIV shovvs the presence of a pol gene in one open reading frame [Ratner, L sl ai., Nature, 313. 277 (1985)]. Amino acid sequence homology provides evidence that the ool sequence encodes reverse transcriptase, an endonuclease and an HIV protease (Toh, H. et al.. EM8O J. 4,1267 (1985); Power, M.D. sl al., Science, 231.1567 (1986); Paart, LH.aLal-, Nature 322, 351 (1987)].

[0003] 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.

[0004] US-A-3526621 discloses substituted 3,1-benzoxa2in-2-ones having relaxing activity on striated muscular flbre and anticonvulsant, tranquilizing and anti-Parkinsonian activity.

[0005] EP-A-93922 discloses benzoxazln-2-ones having antithrombotic activity.

[0006] Compounds of formula I, as herein defined, are cflsclosed. These compounds are useful In the inhibition of HIV reverse transcriptase (and its resistant vārieties), 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 vvith other antivirals, anti-infectives, immunomodulators, antibiotics or vaocines. Methods of treating AIDS, methods of preventing infection by HIV, and methods of treating infection by HIV using compounds of formula II are also disclosed.

[0007] The present invention provides the use of a compound of formula II:

wherein

X is halo;

X^I is trihalomethyl; pentaholoethyl; C2.5 alkyl;

Cj.5 alkynyl;

Cg.5 cycloalkyl; orphenyl, naphthyl, tetrahydronaphthyl, biphenyl, phenethyl, anthryl or acenaphthyl; Ζ is 0 or S;

Ris (a) C-ļ-g aikyl, unsubstituted or substituted vvith A, and A is halo, Cķg cycioaikyl, CN, hydroxy, C^ aikoxy, (¼. 4 aikyny(-C_M aikoxy, phenoxy, naphthoxy, tetrahydronaphthoxy, biphenoxy-, phenethoxy, anthroxy, acenaphthoxy, Cļ^ alkylamaino-C.,.₂ alkyl, heterocycle, or phenylthio, naphthylthio, tetrahydronaphthyithio, biphenyithio, phenethylthio, anthrylthio or acenaphthyithio; vvherein heterocyde means a stable 5- to 7-membered monocyclic or stable 8- to 11 -membered bicyclic heterocycfic ring vvhich is either unsaturated, partiaily unsaturated or saturated, and vvhich consists of carbon atoms and from one to four heteroatoms selected from N, O and S, and vvherein the nitrogen and sulfur heteroatoms may optionally be oxidized, and including any bicyclic group in vvhich any of the above-defined heterocydic rings is fused to a benzene ring;

(b) C₂^ alkenyl, unsubstituted or substituted vvith (i) A, or (ii) phenyl, naphthyl, tetrahydronaphthyl, biphenyl, phenethyl, anthryl or acenaphthyl, unsubstituted or substituted vvith A;

(c) C₂.5 alkynyl, unsubstituted or substituted with (i) A, or (ii) phenyl, naphthyi, tetrahydronaphthyl, biphenyl, phenethyl, anthryl or acenaphthyl, unsubstituted or substituted vvith A; or (d) C3.4 cycloalkyl, unsubstituted or substituted vvith (i) A, or (ii) phenyl, naphthyl, tetrahydronaphthyl;, biphenyl, phenethyl, anthryi or acenaphthyl, unsubstituted or substituted vvith A, or a pharmaceutically acceptable salt thereof, for the preparation of a medicament for inhibiting HIV reverse transcriptase, for preventing infection of HIV, for treating infection by HIV or for treating AIDS or ARC.

[0008] This invention also eneompasses a pharmaceutical composition comprising a compound of formula III vvith the proviso that X¹ and R are not both unsubstituted C₁^ alkyl.

[0009] The present invention also provides a compound of formula I:

vvherein:

X is halo,

X^I is trihalomethyi, or pentahaloethyl; ZisO;

Ris (a) C-|_a alkyl, unsubstituted or substituted vvith A, and A is halo, C^e cycloalkyl, CN, hydroxy, C_V4 alkoxy, (¼. ₄ alkynyl-Cļ_4 alkoxy, phenoxy, naphthoxy, tetrahydronaphthoxy, biphenoxy, phenethoxy, anthroxy, acenaphthoxy, C-,.4 alkyfcarbonyl, nitro, dl(C-,_₂ alkyl)amino, alkylamino- Cļ.₂ aikyl, heterocycle, or phenylthio, naphthylthio, tetrahydonaphthylthio, biphenylthio, phenethylthio, anthrytthio or acenaphthylthio; wherein heterocycle means a stable 5- to 7-membered monocycilc or stable 8- to 11-membered bicyclic heterocycfic ring vvhich is either unsaturated, partially unsaturated or saturated, and which consists of carbon atoms and from one to four heteroatoms selected from N, 0 and S, and wherein the nitrogen and sulfur heteroatoms may optionally be oxidized, and including any bicyciic group in vvhich any of the above-defined heterocydic rings is fused to a benzene ring;

(b) C₂.₄ alkenyi, unsubstituted or substituted vvith (i) A,or (ii) phenyl, naphthyl, tetrahydronaphthyl, biphenyl, phenethyi, anthryl oracenaphthyl, unsubstituted or substituted vvith A;

(c) 02.5 alkynyl, unsubstituted or substituted with (i) A. or (ii) phenyi, naphthyl, tetrahydronaphthyl, biphenyl, phenethyl, anthryl or acenaphthyl, unsubstituted or substituted vvith A; or (d) 0^4 cycloalkyl, unsubstituted or substituted vvith

i) A, or ii) phenyi, naphthyl, tetrahydronaphthyl, biphenyl, phenethyl, anthryl or acenaphthyl, unsubstituted or substituted with A, or a pharmaceutically acceptable salt thereof.

[0010] Preferred compounds include Compounds 37.2, 4, 2, 5 and 24 of Table I belovv, in order of descending degree of preference. These compounds have the follovving structure:

Compound 37.2:

[0011]

Cl (-) (-) 6-Chloro-4-cyctopropyiethynyl-4-trifluoromethyl'1,4-dihydro-2H-3,1-benzoxazin-2-one, the most preferred;

Compound 4:

[0012]

Cl (-) [0013] (-) 6-chloro-4-phenylethynyl-4-trifluoromethyl-1.4'dihydro-2H-3,1-benzoxazin-2-one;

Compound 2:

[0014]

[0015] (4/-) 6-chloro-4-(2-cyanophenyl)ethynyl-4-(1,1,1 -trifluoromethyl)-1,4-dihydro-2H-3,1 -benzoxazin-2-one;

Compound 5:

[0016]

[0017] (4/-) 4-( 1 -chloro-1,1 -difluoromethyl)-4-(2-phenyl-ethynyl)-6-chloro-1,4-dihydro-2H-3,1 -benzoxazin-2-one;

Compound 24:

[0018]

(+/-) 4-(2-[dlmethylaminomethylļethynyl)-4-trifluoromethyl-6-chloro-1,4-dihydro-2H-3,1 -benzoxazin-2-one; or a pharmaceutically acceptable salt thereof.

[0019] The compounds of the present invention are specifically illustrated in Tables I and II below:

TABLE I

TABLE Ϊ (Cont'd)

measured in nM or nanomoles/liter.

TABLE I (Cont’d)

TABLE i (Cont'cB

tt σ>

« o

o o

IO

Z c

IO

O z

c o

CO o

o o_ co* hO

O

O in to

O

CO

IO

O

Λ

CM

O

O

O

O

IO

O

O

O

CO

CM

TABLE I (Cont'd)

co Γ- PIO co co co o o CO ·Μ· CM CM

CM CO CO CO o

u_ o

O

I

o

I

CT

U.

o

o

CM o

o o

n

X o

α c

o

QE o

O co

U5

CO ο

ιη

c ο

ιη

C

Ο ιη co_ ς\ί

C ιη

Τ³

C ο

ω j

co <

ίο ο

ο_ ιη

CM 'Γ

CJ ιη

Ο ιη

Μ·

Ο

Ο

Ο_ ιη

CJ

Ο (Ο

150- 55 3,650

ο in a

«

O o

c o

o ©

0' co

Λ

o in ©

co 'T

O ’Ā

O

A m

O τΑ

0		0
CM	CM	10
CM	d	10

TABLE I (Cont’d)

ocl «

ιό in co © S cm -MCO CO 't cm CM

M- UJ CO T

CM CM Ο O

CM CM CM CM

PJ

LL

O co

LL

O co

LL n

LL

O o

CM r

II

Tf

CM in

CM

X

co rCM CM

ΙΌ

Ο «

Ο ο

c ο

ο

ΙΌ

TABLE I (Cont'd)

C ο

ΙΌ

<=1 <

Τ3

C □

Ο

α.

ε σ

Ο

CM <Ν

Ο <Μ in «

O

O o

ΙΛ ε

•o

c o

o co c

o o

o o

o

CO

A o

o in co

O

O

O θ’ o

co

Λ o

in co

O

O

O

O

O

CO

Λ

CM m

o o

co in

TABLE I (ConCd)

r- co r- r*CM CM liī co CM CM •m· in co co

I

-r- CM in m

CO Γ'· CO 00

ocl

Ό c

□ o

£X

E o

o co co

Tt co in co co co <o

Τ3

C

Ο ω

Ξ <

Η ιη σ>

«

Ο

Ο ο

ίο

C +

ιη co

η

LL

α:|

Ό

C □

Ο

α.

Ε ο

Ο

CM

Ι<

c ο

ο φ

OT <η _φ

CL ε

(0

X

UJ φ

.C

Τ3

Φ σ

'>

ο ία.

(Λ

Ό

C α

ο

α.

c ο

φ φ

Τ3

Φ £

Ο +

TABLE II

TABLE ĪI (Cont'd) in σι

[0020] The compounds of the present invention may have asymmetric centers and may occur, except vvhen specifically noted, as racemates, racemic mixtures or as individual diastereomers, or enantiomers, vvith ail isomeric forms being included in the present invention. The term (+/-) is intended to encompass (+) optical isomers or (-) optical isomers or mixtures thereof.

[0021] When any variable (e.g., R) 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.

[0022] As used herein except vvhere noted, ’alkyl' is intended to include both branched- and straight-chain satu18 rated aliphatic hydrocarbon groups having the specified number of cartoon atoms; alkenyl is intended to cover both branched- and straight Chain alkyl groups vvith at least one carbon-carbon double bond; alkynyl is intended to cover both branched- and straight Chain alkyI groups vvith at least one carbon-carbon triple bond. Halogen or halo as used herein, means fluoro, chloro, bromo and iodo.

[0023] As used herein, vvith exceptions as noted, aryl is intended to mean phenyl, naphthyl, tetrahydronaphthyl, biphenyl, phenanthryļ anthryl or acenaphthyl, [0024] The term heterocycle or heterocyclic, as used herein except vvhere noted, represents a stable 5- to 7-membered monocyclic or stable 8- to 11 -membered bicyc)ic heterocyciic ring vvhich is either saturated, partialiy unsaturated or unsaturated, and vvhich consists of carbon atoms and from one to four heteroatoms selected from the group consisting of N, 0 and S, and vvherein the nitrogen and sulfur heteroatoms may optionally be oxidized, and including any bicyclic group in vvhich 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 vvhich results in the creation of a stable structure. Examples of such heterocyclic elements include piperidinyl, piperazinyl, 2-oxopiperazinyi, 2-oxopiperidinyl, 2-oxopyrrolidinyl, 2-oxoazepinyl, azepinyl, pyrroiyl, 4-piperidonyl, pyrrolīdinyl, pyrazolyl, pyrazolidinyl, imidazolyl, imidazolinyl, imidazolidinyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, oxazolyl, oxazolidinyl, isoxazotyl, isoxazolidinyl, morpholinyl, thiazolyl, thiazolidinyl, isothiazolyl, quinuclidinyl, isothiazolidinyl, indolyi, quinolinyl, isoquinolinyl, benzimidazolyl, thiadiazoyi, benzopyranyl, benzothiazolyl, benzoxazolyl, furyl, tetrahydrofuryl, benzofuranyl, tetrahydropyranyl, thienyl, benzothienyl, thiamorpholinyl, thiamorpholinyl sulfoxide, thiamorpholinyl sulfone, and oxadiazolyl.

[0025] The compounds of the present invention can be synthesized by the follovving methods.

V

SCHEME I

Cl

Cl pivaloyl chloride methylene chloride aaueous Na₂CO₃ NH₂ ²³

HN

1. 2 eq. n-BuLi/THF/O°C ^Cl

2. CF₃COOEt/0°C

3. 3NHCI/reflux

CF₃ nh₂

R-MgBr

THF/0°C to RT

1,1’-carbonyldiimidazole

THF/50°C

[0026] In the synthesis of the benzoxazines of the present invention, the general method typically involves cyclization on a benzene nucleus as a final step. See Scheme I. The amino group of parachloroaniline is first protected vvith, e.g. pivaloyl chloride, to give 2. Other less preferable amino protecting groups include t-butoxycarbonyl, acetate or isovaleroyl groups. About 2 equivalents of an alkyllithium are then reacted with 2, preferably n-butyl lithium. No other organo metallic compounds are suitable for this metalation step. Subsequently, reaction with CF₃COOEt followed by quenching gives 3.

[0027] The synthesis of the tertiary carbinol 4 follovvs, by Grignard addition at the ketone of 2. The Grignard reaģent must be a salt of a divalent cation, e.g. Mg** or Zn**. Monovalent cations are found unsuitable, such as Li* or Na*. Suitable solvents include but are not limited to THF or ether. A wide range of temperature conditions are allovved betvveen about 0“C and about room temperature.

[0028] Ring closure to producē the compounds of the present invention 5 is accomplished vvith condensing aģents such as 1_ir-carbonyldiimidazole, phosgene, dimethylcarbonate, diphenylcarbonate, ordi-(paranitrophenyl)carbonate. Cyclization can be accomplished vvith any of these compounds, as vvell as a wide variety of others.

[0029] A specific instance of Scheme I is provided in Scheme IA. It charts the synthesis of L-741,211, vvhich is a racemate of Compound 37.2, as further provided in Example 6.

SCHEME IA

Cl t-BuCOCI

NH₂

NaOH/H₂O

CHCI₃

Cl (100%)

O.

NH

1.2 n-BuLi/THF -78°C->0^QC/1 h

2. EtOOCCF₃/0°c

3. 3N HCI reflux (60%)

(L-741,211)

SCHEME Π

[0030] Scheme II provides one method for derivatizing acetylene substitutions at the 4-position of the benzoxazine nucleus. By way of illustration, Compound S is metalized, then a zinc salt is added. In the Heck reaction the catalyst tetrakis (trīphenylphosphine)palladium(0) complexed vvith Cul is employed to give 7.

SCHEME ΙΠ

Pyrrolidine, paraformaldehyde

Cul

[0031] Scheme III illustrates substitution of a 4-acetylene group vvith an N-containing heterocycle. The Mannich reaction involves a condensation reaction of formaldehyde vvith the heterocycle, e.g. pyrrolidine. Substitution on the termina! carbon follovvs in the presence of Cul as catalysL

SCHEME iv

Et₃N/4-DMAP/CH₂CI₂

1. Chromatography on

Silica Gel_

2. K₂C03/H₂0/2-propanol [0032] Scheme IV illustrates the resolution of optical isomers of the compounds of Formula. I or Formula II. In this example, (-) camphanic acid is the resolving aģent A wide variety of other resolving aģents are suitable, including Q.methyl mandelic acid chloride or Mosheris reaģent It is apparent to the skilled artison how to separate such isomers. [0033] Scheme IV A is specificaify adapted to the resolution of L-741,211 in the isolation of L-743,726. See Scheme

IV A, and Example 6.

SCHEME P/A

(-)-Camphanyl chloride

4-DMAP/Et₃N/CHCI₃

L-743,726 Compound 37.2

SCHEME V

PCI₅ ^Cl t-BuOK/DMSO _ (42%) [0034] Cyclopropyl acetylene is prepared by Scheme V in accordance vvith published procedures, e.g. C. E. Hudson et al., J. Am. Chem. Soc. 94.1158 (1972) and W. Schoberth et al., Synthesis, 703 (1972).

[0035] The compounds of this invention are 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 povverful antiviral compounds. Furthermore, the compounds of this invention are useful in establishing or determining the binding site of other antivirais to HIV reverse transcriptase, e.g., by competitive inhibition. Thus the compounds of this invention are commercial products to be sold for these purposes.

[0036] The compounds of the present inventions are usefui in the inhibition of H1V reverse transcriptase, the prevention or 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 HIV 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 usefui in treating infection by HIV after suspected past exposure to HIV by e.g., blood transfusion, exchange of body fluīds, bites, accidental needle stick, or exposure to patient blood during surgery.

[0037] The particular advantage of the compounds of this invention is their potent inhibition against HIV reverse transcriptase rendered resistant to other antivirals, such as L-697,661, vvhich is 3-([(4,7-dichloro-1,3-benzoxazol-2yl)methyl]-amino)-5-ethyl-6-methyl-pyridin-2(1 H)-one; or L-696,229, vvhich is 3-[2-( 1,3-benzoxazol-2-yl)ethyl]-5-ethyl-6methyl-pyridin-2(1H)-one; or AZT.

[0038] For these purposes, the compounds of the present invention may be administered orally, parenterally (including subcutaneous injections, intravenous, intramuscular, intrasternal injection or infusion techniques), by inhalation spray, or rectally, in dosage unit formulations containing conventional non-toxic pharmaceutically-acceptable carriers, adjuvants and vehicles.

[0039] Thus, in accordance vvith 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.

[0040] 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.

[0041] VVhen administered orally as a suspension, these compositions are prepared according to techniques vvellknovvn in the art of pharmaceutical formulation and may contain microcrystalline cellulose for imparting bulk, alginic acid or sodium alginate as a suspending aģent, methylcellulose as a viscosity enhancer, and svveeteners/flavoring aģents knovvn in the art. As immediate reiease tablets, these compositions may contain microcrystalline cellulose, dicalcium phosphate, starch, magnesium stearate and lactose and/or other excipients, binders, extenders, disintegrants, diluents and iubricants knovvn in the art.

[0042] 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 benzyI alcohol or other suitable preservatives, absorption promoters to enhance bioavailability, fluorocarbons, and/or other solubilizing or dispersing aģents known in the art.

[0043] The injectable Solutions or suspensions may be formulated according to knovvn art, using suitable non-toxic, parenterally-acceptable diluents or solvents, such as mannitol, 1,3-butanediol, vvater, Ringeris solution or isotonic sodium chloride solution, or suitable dispersing or vvetting and suspending aģents, such as sterile, bland, flxed oils, including synthetic mono- or diglycerides, and fatty acids, including oleic acid.

[0044] VVhen rectally administered in the form of suppositories, these compositions may be prepared by mixing the drug vvith a suitable non-irritating excipient, such as cocoa butter, synthetic glyceride esters or polyethylene glycols, vvhich are solid at ordinary temperatures, but liquidify and/or dissolve in the rectal cavity to reiease the drug.

[0045] The compounds of this invention can be administered orally to humāns in a dosage range of 1 to 100 mg/kg body vveight in divided doses. One preferred dosage range is 0.1 to 10 mg/kg body vveight orally in divided doses. Another preferred dosage range is 0.1 to 20 mg/kg body weight orally in divided doses. For combination therapy vvith nucleoside analogs, a preferred dosage range is 0.1 to 20 mg/kg body vveight for the compounds of this invention administered orally in divided doses, and 50 mg to 5 g/kg body vveight for nucleoside analogs administered oraliy in divided doses. It vvill be understood, hovvever, that the specific dose Ievel and frequency of dosage for any particular patient may be varied and vvill depend upon a variety of fectors including the activity of the specific compound employed, the metabolic stability and length of action of that compound, the age, body vveight, 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.

[0046] The present invention is also directed to combinations of the HIV reverse transcriptase inhibitor compounds vvith one or more aģents usefui in the treatment of AIDS. For example, the compounds of this invention may be effectively administered, vvhether at periods of pre-exposure and/or post-exposure, in combination vvith effective amounts of the AIDS antivirals, immunomodulators, antiinfectives, or vaccines, such as those in the follovving Tabie.

TABLE
Drue Name AL-721	ANTIVĪRALS Manufacturer Ethigen (Los Angeles, CA)	Indication ARC, PGL HIV positive, AIDS
Recombinant Human Interferon Beta	Triton Biosciences (Almeda, CA)	AIDS, Kaposi’s sarcoma, ARC
Acemannan	Carrington Labs (Irving, TX)	ARC (See also immunomodulators)
Cytovene	Syntex	sight threatening CMV
Ganciclovir	(Palo Alto, CA)	peripheral CMV retini tis
d4T Didehydrodeoxy- thymidine	Bristol-Myers (New York, NY)	AIDS, ARC
ddl Dideoxyinosine	Bristol-Myers (New York, ΝΎ)	AIDS, ARC
EL10	Elan Corp, PLC (Gainesville, GA)	HIV infection (See also immunomodulators)

Drug Name Trisodium Phosphonoformate	Manufacturer Astra Pharm. Products, Inc. (Westborough, MA)	Indication CMV retinitis, HIV infection, other CMV infections
Dideoxycytidine; ddC	Hoffman-La Roche (Nutley, NJ)	AIDS, ARC
Novapren	Novaferon Labs, Inc. (Akron, OH) Diapren, Inc. (Roseville, MN, marketer)	HIV inhibitor
Peptide T Octapeptide Sequence	Peninsula Labs (Belmont, CA)	AIDS
Zidovudine; AZT	Burroughs Wellcome (Rsch. Triangle Park, NC)	AIDS, adv, ARC pediatric AIDS, Kaposi's sarcoma, asymptomatic HIV infection, less severe HIV disease, neurological involvement, in combination vvith therapies.

Drue Name Ansamycin LM 427	Manufacturer Adria Laboratories (Dublin, OH) Erbamont (Stamford, CT)	Indication ARC
Dextran Sulfate	Ueno Fine Chem. Ind. Ltd. (Osaka, Japan)	AIDS, ARC, HIV positive asymptomatic
Virazole	Viratek/ICN	asymptomatic HIV
Ribavirin	(Costa Mesa, CA)	positive, LAS, ARC
Alpha Interferon	Burroughs Wellcome (Rsch. Triangle Park. NC)	Kaposi's sarcoma, HIV in combination w/Retrovir

Acyclovir	Burroughs Wellcome	AIDS, ARC, asymptomatic HIV positive, in combination with AZT
Antibody which neutralizes pH labile alpha aberrant Interferon in an immuno-adsorption column	Advanced Biotherapy Concepts Rockville, MD)	AIDS, ARC

Drug Name

L-697,661

Manufacturer Merck (Rahway, NJ)

L-696,229

Merck (Rahway, NJ)

L-735,524

Merck (Rahway, NJ)

Indication

AIDS, ARC, asymptomatic HIV positive, also in combination vvith AZT.

AIDS, ARC, asymptomatic HIV positive, also in combination vvith AZT.

AIDS, ARC, asymptomatic HIV positive, also in combination vvith AZT.

IMMUNO-MODULATORS

Drue Name AS-lOl	Manufacturer Wyeth-Ayerst Labs. (Philadelphia, PA)	Indication AIDS
Bropirimine	Upjohn (Kalamazoo, Ml)	advanced AIDS
Acemannan	Carrington Labs, Inc. (Irving, TX)	AIDS, ARC (See also antivirals)

Drug Name CL246.738	Manufacturer American Cyanamid (Pearl River, NY) Lederle Labs (Wayne, NJ)	Indication AIDS, Kaposi's sarcoma
ELIO	Elan Corp, PLC (Gainesville, GA)	HTV infection (See also antivirals)
Gamma Interferon	Genentech (S. San Francisco, CA)	ARC, in combination w/TNF (tumor necrosis factor)
Granulocyte Macrophage Colony Stimulating Factor	Genetics Institute (Cambridge, MA) Sandoz (East Hanover, NJ)	AIDS
Granulocyte Macrophage Colony Stimulating Factor	Hoeschst-Roussel (Somerville, NJ) Immunex (Seattle, WA)	AIDS

Granulocyte Macrophage Colony Stimulating Factor	Schering-Plough (Madison, NJ)	AIDS AIDS, in combination w/AZT
HIV Core Particle Immunostimulant	Rorer (Ft. Washington, PA)	seropositive HIV

Drue Name IL-2 lnterleukin-2	Manufacturer Hoffman-La Roche (Nutley, NJ) Immunex	Indication AIDS, ARC, HIV, in combination w/AZT
Immune Globulin Intravenous (human)	Cutter Biological (Berkeley, CA)	pediatric AIDS, in combination w/AZT
IMREG-1	Imreg New Orleans, LA)	AIDS, Kaposi's sarcoma, ARC, PGL
IMREG-2	Imreg (New Orleans, LA)	AIDS, Kaposi's sarcoma, ARC, PGL
Imuthiol Diethyl Dithio Carbamate	Merieux Institute (Miami, FL)	AIDS, ARC
Alpha-2 Interferon	Schering Plough (Madison, NJ)	Kaposi's sarcoma w/AZT: AIDS
Methionine- Enkephalin	ΊΓΝΙ Pharmaceutical (Chicago, IL)	AIDS, ARC
MTP-PE Muramy 1-T ripeptide	Ciba-Geigy Corp. (Sununit, NJ)	Kaposi’s sarcoma
Granulocyte Colony Stimulating Factor	Amgen (Thousand Oaks, CA)	AIDS, in combinatioi w/AZT

Drug Name rCD4 Recombinant Soluble Human CD4	Manufacturer Genentech (S. San Francisco, CA)	Indication AIDS, ARC
Rocombinant Soluble Human CD4	Biogen (Cambridge, MA)	AIDS, ARC

Interferon Alfa 2a	Hoffman-La Roche (Nutley, NJ)	Kaposi’s sarcoma AIDS, ARC, in combination w/AZT
SK&F106528 Soluble T4	Smith, Kline & French Laboratories (Philadelphia, PA)	HIV infection
Thymopentin	Immunobiology Research Institute (Annandale, NJ)	HĪV infection
Tumor Necrosis Factor; TNF	Genentech (S. San Francisco, CA)	ARC, in combination w/gamma Interferon

ANTI-INFECTIVES

Drug Name	Manufacturer	Indication
Clindamycin with	Upjohn	PCP
Primaquine	(Kalamazoo, Ml)
Fluconazolec	Pfizer	ciyptococcal
	(New York, ΝΎ)	meningitis, candidiasis

Pastille Nystatin Pastille	Squibb Corp (Princeton, NJ)	prevention of oral candidiasis
Omidyl Eflomithine	Merrell Dow (Cincinnati, OH)	PCP
Pentamidine Isethionate (IM & IV)	LyphoMed (Rosemont, IL)	PCP treatment
Piritrexim	Burroughs Wellcome (Rsch. Triangle Park, NC)	PCP treatment
Pentamidine isethionate for inhalation	Fisons Corporation (Bedford, MA)	PCP prophylaxis
Spiramycin	Rhone-Poulenc Pharmaceuticals (Princeton, NJ)	cryptosporidial diarrhea
Intraconazole- R51211	Janssen Pharm. (Piscataway, NJ)	histoplasmosis; cryptococcal meningitis
Trimetrexate	V/amer-Lambert	PCP
	OTHER
Drus Name Recombinant Human Erythropoietin	Manufacturer Ortho Pharm. Corp. (Raritan, NJ)	Indication severe anemia assoc. with AZT therapy

Drug Name Megestrol Acetate

Total EnteraI Nutrition

Manufacturer

Bristol-Myers (New York, NY)

Nonvich Eaton Pharmaceuticals (Nonvich, ΝΎ)

Indication treatment of anorexia assoc.w/AJDS diarrhea and malabsorption related to AIDS [0047] It will be understood that the scope of combinations of the compounds of this invention vvith AIDS antivirals, immunomodulators, anti-infectives or vaccines is not limited to the list in the above Table, but includes in principle any combination vvith any pharmaceutical composition useful for the treatment of AIDS. For example, a compound of Formula I or Formula II may be suitably administered in combination vvith a nucleoside analog having knovvn biological activity against HIV reverse transcriptase. Appropriate nucleoside analogs are generally chain terminators and include AZT, ddC, ddl, D4T, HEPT and 3^,-fluoro-2',3^,-dideoxythymidine.

[0048] AZT is synthesized by the methods of J.P. Horvvitz et al., J. Org. Chem. 29, 2076 (1964); R.P. Glinski et al., J. Org. Chem. 38.4299 (1973); C.K. Chu et al., Tetrahedron Letters 29.5349 (1988). Application of AZT as a therapeutic drug in the treatment of AIDS is disclosed in U.S. 4,724,232.

[0049] The compound ddC is synthesized by the methods of J.P. Horvvitz et al., J. Org. Chem. 32. 817 (1967); R.

Marumoto, Chem. Pharm. Buli. 22. 128 (1974); andT.-S. Lin et al., J. Med. Chem. 30, 440 (1987).

[0050] D4T is synthesized by the methods of Herdevvijn, P. et al., J. Med. Chem. 30. 1270 (1987).

[0051] HEPT is synthesized by the methods of Miyasaka, T. et. al. J. Med. Chem. 22 2507 (1989); and A.

Rosowsky, J. Med. Chem. 24,1177 (1981). The synthesis of ddC, ddl and AZT are also described in EPO 484071. [0052] The compound 3'-fluoro-2',3‘-dideoxythymidlne is synthesized by the procedures of Herdevvijn, P. et al., J. Med. Chem. 30, 1270 (1987). The compound L-735,524 is N-(2(R)-hydroxy-1(S)-indanyl)-2(R)-phenylmethyl-4-(S)hydroxy-5-(1-(4-(3-pyridylmethyl)-2(S)-N'-(t-butylcarboxamido)-piperazinyl))-pentaneamide, or pharmaceutically acceptable salt thereof. L-697,661 or '661' is 3-([4,7-dichloro-1,3-benzoxazol-2-yl)methyl]-amino)-5-ethyl-ethyl-6methyl-pyridin-2(1 H)-one; L-696,229 is 3-(2-(1,3-benzoxazol-2-yl)-ethyl]-5-ethyl-6-methyl-pyridin-2(1 H)-one. The synthesis of L-697,661 and L-696,229 is described in EPO 484071, and EPO 462800, both herein incorporated by reference.

[0053] Preferred combinations are simuitaneous, intermittent, or aiternating treatments of L-743,726 vvith or vvithout an inhibitor of HIV protease. An optional third component in the combination is a nucleoside inhibitor of HIV reverse transcriptase, such as AZT, ddC or ddl. A preferred inhibitor of HIV protease is L-735,524. Other preferred inhibitors of HIV reverse transcriptase include L-697,661. These combinations may have synergistic effects on limiting the spread of HIV. Preferred combinations include the following: (1) L-743,726 vvith L-735,524, and, optionally any of L-697,661, AZT, ddl orddC; (2) L-743,726 and any of L-697,661, AZT, ddl orddC. Pharmaceutically acceptable salts of these combinations are also included.

ΕΧΑΜΡΙΕ 1.

(+/-) 4-(1,1,1.-trifluoromethvh-4-f1-buten-4-yh-6-chloro-1.4-dlhvdro-2H-3.1-benzoxazin-2-one: (Compound 15)

Steo A: N-(4-chloroohenyn-2.2-dimethvtorooanamide [0054] To a 5L 3 necked round bottomed flask vvith an overhead stirrer vvas added 4-chloroaniline (127.57 g, 1 mole), 1200 mL of CHCI₃, and 1200 mL of saturated aqueous Na₂CO₃ solution. An addition funnel vvas attached to the flask and charged vvith 2,2-dimethylpropanoyl chloride (129 mL, 1.05 mole). The acid chloride vvas added dropwise to the vigorously stirred mixture over 1 h. The resulting mixture was stirred at ambient temperature for an additional 23h. Some of the product separated from the mixture as whrte crystals. These crystals vvere collected by filtration. The filtrate was transferred to a separatory funnel and the layers vvere separated. The chloroform layer vvas vvashed vvith vvater and brine. Drying (MgSO₄), filtration, and.removal of the solvent in vacuo gavē additional product The tvvo portions of prod36 uct were combined and recrystallized from boiling EtOAc-hexanes to give 185.6 g of N-(4-chlorophenyl)-2,2-dimethyl propanamide as a vvhite crystalline solid.

Step Β: l-(2-amino-5-chlorophenyll-2.2.2:trifluoroethanone [0055] To an oven dried, 3L, 3 necked round bottomed flask vvith an overhead stirrer, argon inlet, and a 500 mL oven dried addition funnel vvas added N-(4-chloraphenyl)-2,2-dimethylpropanamide (100 g, 472 mmol) and dry THF (1L). This solution vvas cooled in an ice bath to 0°C and the addition funnel vvas charged vvith n-butyllithium (387 mL of a 2.5 M solution in hexanes, 968 mmol). The n-butyllithium solution vvas added dropvvise to the amide solution slowly over 1 h, maintaining the temperature belovv +5’C. The resulting solution vvas aged at 0°C for 1h during vvhich time an orange precipitate formed. To this mixture vvas added ethyl 1,1,1 -trifluoroacetate (115 mL, 968 mmol), dropvvise over 1 h. The resulting clear solution vvas aged an additional 30 min. The reaction vvas quenched vvith 5% aqueous HCI. The mixture vvas diluted vvith 1L of EtOAc and the layers vvere separated. The organic phase vvas vvashed with brine, dried (MgSO₄), filtered and concentrated in vacuo, to give 160 g of a yellow oil. This material vvas suspended in 1L of 3Ņ aqueous HCI and the solution vvas heated at reflux for 24h. The cooled solution vvas diluted vvith 1L of EtOAc and the mixture vvas made basie vvith concentrated NH₄OH. The layers vvere separated and the organic phase vvas vvashed vvith brine, dried (MgSO₄), filtered, concentrated iņ vacuo. and chromatographed on 1.5 kg of silica gel using 15% EtOAc in hexane as eluant The chromatographed material vvas recrystallized from boiling hexane to give 57 g (54%) of pure 1-(2-amino-5chlorophenyl)-2,2 ,2-trifluoroethanone as bright yellow crystals, mp: 91 -92°C. ¹H NMR (CDCI₃): δ 6.46 (br s, 2H), 6.69 (d, 1H, J=9.2 Hz), 7.32 (dd, IH, J=2.4, 9.2 Hz), 7.70 (d, 1H, J=2.4 Hz).

Step C: M-) 2-(2-Amino-5-chlorophenyn-1.1.1-trifluoro-5-hexen-2-ol [0056] To a 300 mL oven dried 3 necked, round bottomed flask vvith a stirring bar, argon inlet, addition funnel and a reflux condenser vvas added magnesium (turnings, 3.03 g, 125 mmol) and dry THF (75 mL). To this vvell stirred mixture vvas added 4-bromo-1 -butene (12.0 mL, 118.21 mmol) at such a rāte as to maintain a gentle reflux. VVhen the addition vvas complete, the mixture vvas aged 30 min then cooled to 0°C in an ice bath. To this vvell stirred solution vvas added a solution of 1-(2-amino-5-chlorophenyl)-2,2,2-trifiuoroethanone (5.00 g, 22.36 mmol) in THF (35 mL), dropvvise over 30 min. The cooling bath vvas allowed to expire and the mixture vvas stirred 20 h at ambient temperature. The reaction vvas diluted vvith EtOAc and 10% aqueous citric acid. This mixture vvas stirred for 4 h. The layers vvere separated and the organic phase vvas vvashed vvith aqueous NaHCO₃ and brine. Drying (MgSO4), filtration, removal of the solvent in vacuo. and chromatography on 300g of silica gel using 15% EtOAc in hexane as eluant gavē 4.80 g of (+/-) 2-(2amino-5-chlorophenyl)-1,1_>1-trifluoro-5-hexen-2-ol as a yellow solid.

Step D: (+/-} 4-(1.1.1. -trifluoromethvO-4-f 1 -buten-4-yn-6-chloro-1,4-dihvdro-2H-3.1 -benzoxazin-2-one [0057] To a 200 mL round bottomed flask vvith a stirring bar, argon inlet and a reftux condenser was added (+/-) 2(2-Amino-5-chlorophenyl)-1,1,1-trifluoro-5-hexen-2-ol (4.80 g, 17.16 mmol), 1,1'-carbony(diimidazole (13.91 g, 85.81 mmol) and dry THF (75 mL). This mixture vvas heated at 60°C for 18h. The cooled reaction mixture vvas diluted vvith EtOAc and vvashed vvith H₂O (3X 200 mL) and brine (250 mL). Drying (MgSO₄), filtration, removai of the solvent in vacuo. follovved by recrystallization from boiling EtOAc-hexane gavē 3.22g of (+/-) 4-(1,1,1,-trifluoromethyl)-4(1-buten4-yl)-6-chloro-1,4-dihydro-2H-3,1-benzoxazin-2-one as a vvhite crystaliine solid, mp: 165-166°C. ¹H NMR (C0CI₃): δ 1.99 (m, 1H), 2.09-2.40 (m, 3H), 5.00 (d, 1H, J=1.4 Hz), 5.03 (dd, 1H, J=1.4, 7.9), 5.78 (m, 1H), 6.85 (d, 1H, J=8.6 Hz), 7.21 (br s, 1H), 7.35 (dd, 1H, J=2.2, 8.6 Hz), 9.63 (br s, 1H).

EXAMPLE 2 (+/-) 6-Chloro-4-ethynyl-4-(1.1.1-trifluoromethvl)-1.4-dihvdro-2H-3.1-benzoxazin-2-one (Compound 26)

Step A: 2-/2-amino-5-chlorophenvl)-1,1,1 -trifluoro-3-butvn-2-ol [0058] A 500 ml 3-neck round bottom flask fitted vvith an addition funnel, argon inlet, stirring bar and digital thermometer vvas charged with ethynyl magnesium bromide (0.5M in hexane; 268 mL, 134 mmol) then chilled to -78°C. Dropvvise addition of a solution of 1-(2-amino-5-chlorophenyl)-2,2,2-trifluororethanone (6.0 g, 26.8 mmol) in 50 mLTHF vvas completed after 15 minūtes keeping the temperature £ -55’C. The reaction mixture was stirred for 16 h after slowly vvarming to room temperature. The dark red solution vvas quenched at -5°C by dropvvise addition of saturated aqueous ammonium chloride solution (60 mL). Ethyl acetate extraction follovved by vvashes of 10% citric acid, saturated sodium bicarbonate, vvater and brine afforded 8.5 g crude product after drying over sodium sulfate, filtration, and evaporation of solvent. Purification via flash chromatography using 15-20% ethyl acetate : hexane afforded pure 2-(2-amino-5-chlorophenyl)-1,1_l1-trifluoro-3-butyn-2-ol (5 g light brown oil, 75% yield).

Step B: (+/-1 6-Chloro-4-ethynyl-4-(1.1.1-trifluoromethyl)-1.4-dihydro-2H-3.1-benzoxazin-2-one [0059] A THF solution of 2-(2-amino-5-chlorophenyl)-1,1,1-trifluoro-3-butyn-2-ol (5.0 g, 20.0 mmol in 225 mL THF) was treated vvith 1,1'-carbonyldiimidazole (13.0 g, 80.0 mmol) and heated in an oil bath at 60°C for 17 h. The THF vvas removed in vacuo, the residue dissolved in ethyl acetate then vvashed vvith 10% citric acid, sodium bicarbonate, vvater and brine before drying over sodium sulfate. Follovving filtration and evaporation iņ vacuo the crude product vvas isolated (3.6 g) and recrystallized from ethyl acetate : hexane. The product (+/-) 6-chloro-4-ethynyi-4-(1,1,1 -trifluoromethyl)-1,4dihydro-2H-3,1-benzoxazin-2-one vvas isolated as a vvhite crystalline solid (3.22 g, 58.4% yield): mp 226-227’C. ¹HNMR (CDCI3 + trace DMSO): δ 3.16 (s, 1H), 6.98 (d, J=3.3 Hz, 1H), 7.35 (m, 1H), 7.51 (s, 1H), 10.66 (s, 1H).

ΕΧΑΜΡΙΕ 3

Μ-Ί 6-Chloro-4-(1.1,1 -trifluoromethyl)-4-[(3-(1 -pyrrolidinyl1)-1 -propynyl]-1.4-dihydro-2H-3.1 -benzoxazin-2-one (Compound 71 [0060] A dioxane solution of (+/-) 6-chloro-4-ethynyl-4-(1,1,1 -trifluoromethyl)-1,4-dihydro-2H-3,1 -benzoxazin-2-one (150 mg, 0.544 mmol), pyrrolidine (52.2 pL, 0.626 mmol), paraformaldehyde (20.5 mg, 0.681 mmol), acetic acid (31.1 pL, 0.544 mmol) and copper (I) chloride (20.5 mg, 0.207 mmoi in 3.5 ml dioxane) was heated to 50’C in an oil bath for approximately 2 h. The reaction mixture vvas quenched into 2M HCI and extracted vvith ethyl acetate. The aqueous layer vvas neutralized vvith solid potassium carbonate and extracted vvith ethyl acetate (3x). The combined extracts vvere vvashed with vvater and brine before drying over sodium sulfate to afford 140 mg crude product. Chromatographic purification on silica gel and recrystallization from ethyl acetate : hexane afforded crystalllne (+/-) 6-chloro-4(1,1_I1trifIuoromethyl)-4-[(3-{1-pyrrolidinyl))-1-propynyl]-1,4-dihydro-2H-3,1-benzoxazin-2-one (89mg, 46% yield): MP 160-161’C (dec). ¹H-NMR (COCI3): δ 1.85-1.89 (m, 4H), 2.68-2.71 (m, 4H), 3.67 (s, 1H), 6.88 (d, J= 8.55 Hz, 1H), 7.40 (dd, J= 2.19, 8.54 Hz, 1H), 7.55 (s, 1H), 9.45 (s, 1H).

<+/-} 6-Chloro-4-(2-cvanophenvl1ethvnvl-4-(1.1.1 -trifluoromethvll-1.4-dihvdro-2H-3.1 -benzoxazin-2-one (Comoound 21 [0061] A solution of 6-Chloro-4-ethynyl-4-(1,1,1-trifluoromethyl)-1,4-dihydro-2H-3,1-benzoxazin-2-one(138mg, 0.5mmol)in 3 mL of dry THF was stirred at -78’C. To this solution vvas added 0.4 mL (1.0 mmol) of n-butyllithium, 2.5 M in hexane. The anion was aged for 10 minūtes at -78°C then 1 mL of ZnCIjO M in ether) solution vvas added. The reaction mixture vvas allowed to stir at -78°C for 15 minūtes, the ice bath vvas removed and the mixture slowly vvarmed to 0’C over 30 min. To the reaction mixture vvas added a solution of 2-iodobenzonitriie (149 mg, 0.65 mmol) in 2 mL THF, follovved by tetrakis(triphenylphosphine)palladium(0) (56 mg, 0.05 mmol). The reaction vvas allovved to vvarm to r.t. and continued to stir over 15 hours. The reaction mixture vvas quenched vvith 10 mL of 2ļM HCI, extracted vvith 2x200 mL EtOAc and the combined extracts vvere vvashed vvith H₂O, brine and dried over MgSO₄. The solvent vvas removed to give 195 mg of an oil which vvas flashed chromatographed on silica gel (20% EtOAc in hexane) to afford 60 mg of the unreacted starting material and 35 mg of the coupled product The latter was triturated vvith ether to yield 25 mg of (+/) 6-Chloro-4-(2-cyanophenyi)ethynyl-4-(1,1,1-trifluoromethyl)-1,4-dihydro-2H-3,1-benzoxazin-2-one. mp: 245-246’C FAB. MS M+1=377 m/e. ¹H NMR (C0Ci₃): δ 6.82-6.85 (d, J=8.5 Hz, 1H); 7.40-7.44 (dd, J=2.1, 8.5 Hz, 1H); 7.56-7.79 (m, 5H); 8.00 (s, 1H).

EXAMPLE4 (+/-14-/1-Chloro-1.1-difluoromethvn-4-(2-Qhenvlethvnvn-6-chloro-1.4-dihvdro-2H-3.1-benzoxazin-2-one (Comoound 5)

Step A: 1-(2-amino-5-chlorophenvt1-2-chloro-2.2-difluoroethanone [0062] To an oven dried, 300 mL, 3 necked round bottomed flask vvith a magnetic stirring bar, argon inlet, and a 100 mL oven dried addition funnel vvas added N-(4-chlorophenyl)-2,2-dimethyl-propanamide (10 g, 47.2 mmol) and dry THF (100 mL). This solution vvas cooled in an ice bath to 0°C and the addition funnel vvas charged with n-butyllrthium (38.7 mL of a 2.5 M solution in hexanes, 96.8 mmol). The n-butyllithium solution vvas added dropvvise to the amide solution slowly over 1 h, maintaining the temperature belovv +5’C. The resulting solution was aged at 0’C for 1h during vvhich time an orange precipitate formed. To this mixture vvas added ethyl 1-chloro-1,1-difluoroacetate (10.2 mL, 96.8 mmol), dropwise over 15 min. The resulting clear solution vvas aged an additional 30 min. The reaction vvas quenched vvith 5% aqueous HCl. The mixture vvas diluted vvith 1L of EtOAc and the layers vvere separated. The organic phase was vvashed vvith brine, dried (MgS0₄), filtered and concentrated in vacuo. to give 160 g of a yellow oil. This material vvas suspended in 200 mL of 3N aqueous HCl and the solution vvas heated at reflux for 24h. The cooled solution vvas diluted vvith 500 mL of EtOAc and the mixture vvas made basie vvith concentrated NH₄OH. The iayers vvere separated and the organic phase vvas vvashed vvith brine, dried (MgSO₄), filtered, concentrated in vacuo, and chromatographed on 350 g of silica gel using 15% EtOAc in hexane as eluant. The chromatographed material vvas recrystallized from boiling hexane to give

5.5 g of pure 1-(2-amino-5-chlorophenyl)-2-chloro-2,2-difluoroethanone as bright yellow crystals, mp: 55-56°C. ¹H NMR (CDCI₃): δ 6.43 (brs, 2H), 6.69 (d, 1H, J=9.0 Hz), 7.31 (dd, 1H, J=2.4, 9.0 Hz), 7.80 (d, 1H, J=2.4 Hz).

Step B: f+/-1 2-(2-amino-5-chloroohenylM-phenyl-1-chloro-1.1-difluoro-3-butyn-2-ol [0063] To a 100 mL, 3 neeked, oven dried round bottomed flask, vvith a stirring bar, argon inlet, reflux condenser, and a septum vvas added ethynyl benzene (2.13 g, 20.83 mmol), dry THF (50 mL) and ethyl magnesium bromide (6.94 mL of a 3.0M solution in ether). This mixture vvas aged 2h at ambient temperature then a solution of 1 -(2-amino-5-chlorophenyl)-2-chloro-2,2-difluoroethanone (1.00 g, 4.17 mmol) in THF (6 mL) vvas added vvith a syringe. The resulting orangered solution vvas stirred at ambient temperature for 21.5h. The reaction vvas quenched by addition of 1fcļ HCl (50 mL) then diluted vvith EtOAc. The solution vvas then made basie vvith concentrated NH₄OH and the layers vvere separated. The organic phase vvas vvashed vvith vvater and brine. Drying (MgSO₄), filtration, removal of the solvent in vacuo, and chromatography on silica gel using 20% EtOAc in hexane as eluant gavē 1.02 g of (+/-) 2-(2-amino-5-chlorophenyi)-4-phenyi-1 -chloro-1,1 -difluoro-3-butyn-2-ol as an off vvhite solid.¹H NMR (C0CI₃): δ 4.42 (br s, 2H), 5.10 (br s, 1H), 6.65 (d, 1H, J=8.5 Hz), 7.15 (dd, 1H, J=2.4, 8.5 Hz), 7.38 (m, 3H), 7.55 (m, 2H), 7.70 (d, J=2.4 Hz).

Step C: (+/-} 4-i 1 -Chloro-1.1 -dffluoromethvn-4-(2-ohenvl-ethvnyn-6-chloro-1.4-dihvdro-2H-3.1 -benzoxazin-2-one [0064] To a 100 mL round bottomed flask vvith a stirring bar, reflux condenser, and an argon inlet was added (+/-) 2-(2-amino-5-chlorophenyl)-4-phenyl-1 -chloro-1,1 -difluoro-3-butyn-2-ol (0.81 g, 2.37 mmol), dry THF (25 mL), and 1,1'cartjonyldiimidazole (1.919 g, 11.84 mmol). This solution vvas heated at 60°C for 20h. The cooled reaction mixture vvas diluted vvith EtOAc and vvashed vvith O.Sfcļ HCl H₂O, and brine. Drying (MgSO₄), filtration, and removal of the solvent in vacuo gavē 890 mg of an oil. This material vvas chromatographed on 80 g of silica gel using 20% EtOAc in hexane as eluant The chromatographed material vvas recrystallized from boiling EtOAc-hexanes to give 507 mg, (58%) of (+Z-) 4(1 -chloro-1,1-difluoromethyl)-4-(2-phenylethynyl)-6-chloro-1,4-dihydro-2H-3,1-benzoxazin-2-one as vvhite needles, mp: 154-155°C. ¹H NMR (CDCI₃): δ 6.89 (d. 1H, J=8.4 Hz), 7.35-7.48 (m, 4H), 7.56 (m,2H), 7.64 (brs, 1H), 9.19 (brs, 1H).

ΕΧΑΜΡΙΕ.5 f-1 6-Chloro-4-Dhenvlethvnvi-4-trifluoromethyl-1.4-dihvdro-2H-3.1-benzoxazin-2-one (Compound 41

Steo A: 2-f2-Amino-5-chlorophenvD-4-phenvl-1.1.1 -trifluoro-3-butyn-2-ol [0065] A solution of lithio phenylacetylide, prepared from 4.83 mL of phenylacetylene (0.044 mol) and 17.2 mL of a

2.5 Ņ solution of n-butyllithium in hexane (0.043 mol) in 50 mL of THF at -78°C, vvas treated with 11.4 g of magnesium bromide etherate (0.044 mol) over 5 minūtes. The mixture vvas allovved to warm to -20^eC and stirring under argon was continued for 30 minūtes. The mixture vvas then cooled to -60°C and a solution containing 2.5 g (0.011 mol) of 1-(2amino-5-chloro)-2,2,2-trifluoromethylethanone, previously complexed vvith an equivaient (2.8 g, 0.011 mol) of magnesium bromide etherate in 25 mL of THF, vvas added. The reaction mixture vvas allovved to stir at 15° for one hour before being cooled to 0°C and treated dropvvise vvith a mixture of 30 mL each of saturated aqueous ammonium chloride solution and vvater. The mixture vvas extracted vvith 2 x 100 mL portions of ethyl ether, the combined organic phases vvere vvashed vvith brine and dried over MgSO4. Removal of the drying aģent and solvents left 6 g of an oil vvhich vvas flash chromatographed on silica gel, eluting vvith 20% EtOAc in hexane, to afford 2.5 g of 2-(2-amino-5-chlorophenyl)-4-phenyl-1,1,1-trifiuoro-3-butyn-2-ol.¹H-NMR (CDCI3>: δ 4.63 (brs, 3H), 6.69 (d, J=3.5 Hz, 1H), 7.15 (d, J= 2 Hz, 1H), 7.17 (d, J=2 Hz, 1H), 7.35-7.44 (m, 3H), 7.53-7.56 (m, 2H), 7.66 (d, J=2 Hz, 1H). FAB MS M+H = 326 m/e.

Step B: (±1 6-Chloro-4-phenvlethvnvi-4-trifluoromethvi-1.4-dihvdro-2H-3.1-benzoxazin-2-one (Compound 121 [0066] A solution of 2-(2-amino-5-chlorophenyl)-4-phenyl-1,1,1-trifiuoro-3-butyn-2-ol (2.0 g, 6.1 mmol) and 11.0 g (12.0 mmol) of 1,1 '-carbonyldiimidazole in 300 mL of dry THF was stirred under argon at 55°C for 24 hours. The solvent vvas removed on a rotary evaporator and the residue vvas partitioned betvveen 200 mL of ether and 400 mL of vvater. The layers vvere separated and the aqueous extracted once more with ether. The combined ether extracts vvere washed vvith 2 x 200 mL 10% citric acid and then vvith brine before drying over MgSO₄. Removal of the drying aģent and solvent provided 1.5 g (70%) of the crude title compound as an oil. Trituration vvith ether-hexane afforded 875 mg of (±) 6chloro-4-phenylethynyl-4-trifluoromethyl-1,4-dihydro-2H-3,1 -benzoxazin-2-one as a vvhite solid, partial meit at 137°, clearat 147’C. ¹H-NMR (CDCķ): δ 6.92 (d, J=8 Hz, 1H), 7.30-7.49 (m, 4H), 7.58-7.65 (m, 3H), 8.99 (s, 1H).

Step C: 6-Chloro-1 -(1 S)-camphanoyl-4-phenvlethvnvl-4-trifluoromethyl-1.4-dihvdro-2H-3.1 -benzoxazin-2-one [0067] To a solution containing (±) 6-chloro-4-phenylethynyl-4-trifluoromethyl-1,4-dihydro-2H-3,1-benzoxazin-2one (2.24 g, 6.37 mmol), 4-dimethylaminopyridine (0.10 g, 0.8 mmol), and (-) camphanic acid chloride (2.07 g, 9.55 mmol) in 60 mL of dry dichloromethane, stirred under argon in an ice bath, vvas added triethylamine (2.22 mL, 15.9 mmol). The cooling bath vvas removed and the reaction was allovved to proceed at room temperature. When the reaction vvas complete by thin layer chromatography (SiO₂, 4% EtOAc in CHCķ), the solution vvas diiuted with 200 mL of CHCI3 and vvashed tvvice with 10% citric acid, then vvith brine. Upon drying (MgSO₄) the solvent vvas removed on a rotary evaporator and the foamy reside vvas subjected to flash chromatography, eluting vvith CHCķ There vvas obtained 575 mg of diastereomer I of 6-chloro-(1S)-camphanoyl-4-phenylethynyl-4-trifluoromethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one as an oil, ¹H-NMR (CDCķ): δ 0.85 (s, 3H), 1.08 (s, 3H), 1.22 (s, 3H), 1.73-1.85 (m, 1H), 1.92-2.08 (m, 1H), 2.50-2.67 (m, 2H), 7.30-7.79 (m, 8H). This vvas follovved by 1.52 g of mixed fractions (diastereomers I and II). Continued elution afforded 680 mg of the slovver-moving diastereomer (II) of the title compound vvhich, upon trituration vvith an ether/hexane mixture, gavē clumps of vvhite needles, mp 177-178.5’C; ¹H-NMR (C0CI₃): δ 0.83 (s, 3H), 1.12 (s, 3H), 1.23 (s, 3H), 1.73-1.86 (m, 1H), 1.93-Ζ06 (m, 1H), 2.50-2.63 (m, 2H), 7.38-7.51 (m, 4H), 7.49-7.62 (m, 2H), 7.72 (d, J=9 Hz, 1H), 7.76 (d,J=2 Hz, 1H).

[0068] The 1.52 g of isomer mixture from the flash chromatography vvas dissolved in 75 mL of ether, the solution diiuted vvith 50 mL of hexane, and then seeded vvith a crystal of isomer II. Slow crystallization afforded an additional 385 mg of isomer II vvhich vvas recrystallized from ether/hexane to give >96% diastereomerically pure material by HPLC.

Step D: (-1 6-Chloro-4-phenvlethvnvl-4-trifluoromethvl-1,4-dihvdro-2H-3.1 -benzoxazin-2-one [0069] The crystalline diastereomer(ll) of 6-chloro-1-(1S)-camphanoyl-4-phenylethynyl-4-trifluoromethyl-1,2-dihydro-4(H)-3,1-benzoxazin-2-one (53 mg, 0.10 mmol) was dissolved in 8 mL of 2-propanol at 45’C under an atmosphere of argon. To the solution vvas added 0.27 mL of a 10% aqueous solution of K₂CO₃. Stirring vvas continued for 10 min., by vvhich time ali of the starting material had been consumed (TLC, SiO₂, 4% EtOAc in CHCl^. The solution vvas con- centrated in vacuo and the residue taken up in ether. After vvashing vvith 0.1 M HCl and brine, the ether solution vvas . dried (MgSO^, filtered and evaporated in vacuo to an oify solid vvhich vvas purified by SiO₂ chromatography, eluant 5% 2-propanol in hexane. (-) 6-Chloro-4-phenylethynyl-4-trifluoromethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one was obtained as vvhite needles from ethedhexane, m.p. 178-179°C; [a]_D20 = - 92.5° (CHCI₃, c=0.0012 g mL-1); ¹H-NMR (CDCķ): δ 6.87 (d, J=8.5 Hz, 1H), 7.37-7.50 (m, 4H). 7.56-7.63 (m, 3H), 8.60 (s, 1H).

Step E: (+) 6-Chloro-4-phenvlethvnvl-4-trifluoromethvl 1.4-dihvdro-2H-3.1-benzoxazin-2-one fCompound 3) [0070] (+) 6-Chloro-4-phenylethynyl-4-trifluoromethyl-1,4-dihydro-2H-3,1 -benzoxazin-2-one was prepared from the non-crystalllne product of Step C, diasteromer I, in a manner according to Step D: m.p. 178-179’C; [a]D²⁰=+ 87.6’ (CHCķ, c=0.0050 g mL-1; ¹H-NMR(C0Cl3): δ 6.87 (d, J=8.5 Hz, 1H), 7.37-7.50 (m, 4H), 7.56-7.63 (m, 3H), 8.60 (s, 1H).

ΕΧΑΜΡΙΕ 6 (-) 6-Chloro-4-cvclopropvtethvnvl-4-trif1uoromethvl-1.4-dihvdro-2H-3.1 -benzoxazin-2-one(L-743.726, Compound 37.2) and (+) 6-Chloro-4-cvdoDroDvlethvnvl-4-trifluoronriethvl-1.4-dlhydro-2H-3.1-benzoxazin-2-one (L- 743.7251

Step A: 2-(2-Amino-5-chloroūhenvO-4-cvclopropvl-1.1.1 -trifluoro-3-butvn-2-ol [0071] A solution of bromomagnesium cydopropy1acetylide, was prepared from 23 g of cydopropylacetylene (0.348 mol) in 250 mL of THF by dropvvise addition of 116 mL of a 3.0 M solution of ethylmagnesium bromide in ether (0.348 mol) over 1h. This solution was maintained at 0°C for 1h, then at 40*C for 3h. To this solution, recooled to 0’C, 15.56 g of 1-(2-amino-5-chlorophenyl)-2,2,2-trifluoromethylethanone (0.0696 mol), was added as a solid, portionvvise over 5 min. The reaction mixture was allovved to stir at 0’ for 1.5 hours. The reaction vvas quenched at 0’C by dropvvise addition of 700 mL of saturated aqueous ammonium chloride solution. The mixture vvas extracted vvith 2 x 400 mL portions of ethyl acetate, the combined organic phases were vvashed vvith brine and dried over MgSO₄. Removal of the dry40 ing aģent and solvents left a yellow solid. This material vvas recrystallized from boiling hexanes (100 mL final volume) to afford 14.67 g of 2-(2-amino-5-chlorophenyl)-4-cyclopropyl-1,1,1-trifluoro-3-butyn-2-ol. A second crop (2.1g) vvas obtained from concentrating the mother liquors. mp: 153-154°C. ¹H-NMR (COCI3): δ 0.84 (m, 2H), 0.90 (m, 2H), 1.38 (m,1H), 4.50 (br s, 3H), 6.69 (d, J = 8.5 Hz, 1H), 7.13 (dd, J = 2.5, 8.5 Hz, 1H), 7.55 (d, J = 2.5 Hz, 1H).

Step B: (±) 6-Chloro-4-cvclopropvlethynyl-4-trifluoromethyl-1,4-dihvdro-2H-3.1 -benzoxazin-2-one (L-741.2111 [0072] A solution of 2-(2-amino-5-chlorophenyl)-4-cyclopropyl-1,1,1-trifluoro-3-butyn-2-ol (15.00 g, 0.0518 mol) and 41.98 g (0.259 mol) of 1,1'-carbonyldiimidazole in 250 mLof dry THF vvas stirred under argon at55°Cfor24 hours. The solvent vvas removed on a rotary evaporator and the residue vvas partitioned betvveen 500 mL of ethyl acetate and 400 mL of vvater. The layers vvere separated and the aqueous phase vvas extracted once more vvith ethyl acetate. The combined ethyl acetate extracts vvere vvashed vvith 2 x 200 mL of 2% aqueous HCl, saturated aqueous NaHCO₃, and brine. Drying over MgSO₄, filtration, and removal of the solvent in vacuo provided 16.42 g of the title compound as a solid.. Recrystallization from ethyl acetatehexane afforded 12.97 g of analytically pure (±) 6-chloro-4-cyclopropylethynyl-4-trifluoromethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one as avvhite crystals. mp: 178-180°C. ¹H-NMR (CDCI3): 0.85 (m, 2H), 0.94 (m, 2H), 1.40 (m, 1H), 6.81 (d, J = 8.5 Hz, 1H), 7.37 (dd, J = 2.5, 8.5 Hz, 1H), 7.49 (d, J = 2.5 Hz, 1H), 8.87 (brs, 1H).

Step C: 6-Chloro-1-(1S)-camphanovl-4-cvclopropvlethvnvl-4-trifiuoromethvl-1.4-dihvdro-2H-3.1-benzoxazin-2-one [0073] To a solution containing (±) 6-chloro-4-cyclopropylethynyl-4-trifluoromethyl-1,4-dihydro-2H-3,1-benzoxazin2-one (12.97 g, 0.041 mol), 4-dimethylaminopyridine (1.02 g, 0.0083 mol), and (-) camphanic acid chloride (14.22 g, 0.06556 mol) in 350 mL of dry dichloromethane, stirred under argon in an ice bath, vvas added triethylamine (22.84 mL, 0.164 mol). The cooling bath vvas removed and the reaction vvas allovved to proceed at room temperature. After 75 min. the reaction vvas judged complete by thin layer chromatography (SiO₂, 4% EtOAc in CHCI₃), and the solution vvas diluted with 500 mL of CHCLj then vvashed vvith 10% c'rtric acid (2X), water (IX), and brine (1X). Drying (MgSO₄), filtration, and removal of the solvent in vacuo left a colorless foam. This material was triturated vvith 200 mL of boiling hexane. On cooling to room temperature the desired diastereomeric camphanate imide precipitated. The solid was collected on a frit, vvashed vvith a little cold hexanes and dried iQ vacuo to give 7.79g of 6-chloro-1 -(1 S)-camphanoyl-4cyclopropylethynyl-4-trifluoromethyl-1,4-dihydro-2H-3,1 -benzoxazin-2-one as vvhite crystals. mp: 164-165’C, HPLC purity: 99.2% © 254 nm. ¹ H-NMR (CDCI₃): 0.77 (s, 3H), 0.86-0.96 (m, 4H), 1.08 (s, 3H), 1.19 (s, 3H), 1.44 (m, 1H), 1.76 (m, 1H), 1.95 (m, 1H), 2.51 (m,2H), 7.42 (dd, J = Z4.9.0 Hz, 1H), 7.63 (m, 2H).

Step D: (-1 6-Chloro-4-cyclopropylethynyl-4-trifiuoromethyl-1.4-dihydro-2H-3.1-benzoxazin-2-one (L-743.726, Compound 37.2) [0074] 6-chloro-1-(1 S)-camphanoyl-4-cydopropylethynyi-4-trifluoromethyl-1,2-dihydro-4(H)-3,1 -benzoxazin-2-one (7.50 g, 0.01512 mol) vvas dissolved in 150 mL of n-butanol at 60’C under an atmosphere of argon. To this solution vvas added 10 mL of 1Ņ HCl. This solution was maintained at 60’C for 72h. The mixture vvas neutralized with aqueous NaHCO₃ and the n-butanol vvas removed in vacuo. The residue vvas dissolved in 150 mL of THF and treated vvith 50 mL of 2N LiOH for 3h at room temperature. This mixture vvas diluted vvith ethyl acetate and washed vvith tvvo portions of vvater and one of brine. Drying (MgSO₄), filtration and removal of the solvent in vacuo gavē a vvhite solid. This material vvas recrystallized from hot hexane to give 3.43 g of (-) 6-chloro-4-cyclopropylethynyl-4-trifluoromethyl-1,4-dihydro-2H3,1-benzoxazin-2-one as vvhite crystais., m.p. 131-132’C; [aļD²⁰ = - 84.7° (CHCI3, c=0.005 g mL'¹⁾; ’H-NMR (C0CI3): δ 0.85 (m, 2H), 0.94 (m, 2H), 1.40 (m, 1H), 6.81 (d, J = 8.5 Hz, 1H), 7.37 (dd, J = 2.5,8.5 Hz, 1H), 7.49 (d, J = 2.5 Hz, 1H), 8.87 (brs, 1H).

Step E: (+) 6-Chloro-4-cyclopropvlethvnvl-4-trifluoromethvl-1.4-dihvdro-2H-3.1-benzoxazin-2-one fL-743,725) [0075] The mother liquors from Step C above vvere purified by column chromatography on silica gel using 10% ethyl acetate in hexanes as eluant. The pure, undesired diastereomer (a colorless foam) vvas hydroylzed according to Step D. The enantiomeric benzoxazinone, (+) 6-ChloiO-4-cyclopropylethynyl-4-trifluoromethyl-1,4-dihydro-2H-3,1-benzoxazin-2-one, vvas obtained as vvhite crystals. m.p. 131-132’C; [a]D²⁰ = +· 84.4® (CHC^, c=0.005 g mL'¹); ¹H-NMR (CDCI₃): δ 0.85 (m, 2H), 0.94 (m, 2H), 1.40 (m, 1H), 6.81 (d, J = 8.5 Hz, 1H), 7.37 (dd, J = 2.5, 8.5 Hz, 1H), 7.49 (d, J = 2.5 Hz, 1H), 8.87 (brs, 1H).

REVERSE TRANSCRIPTASE ASSAY [0076] The assay measures the incorporation of tritiated deoxyguanosine monophosphate by recombinant HIV reverse transcriptase (HIV RT_R) (or other RT) into acid-precipitable cDNA at the Km values of dGTP and poly r(C) · oligo d(G)i₂.i8- The inhibitors of the present invention inhibit this incorporation.

[0077] The assays vvere carried out in 55 mM Tris (pH 3.2)-30 mM KCI-30 mM MgCIs-l mM dithiothreitol-20 gg of rC:dG₁₂.₁₈ (Pharmacia) per ml-8 mM [³H]dGTP (New England Nuciear)-0.01 % Triton Χ-100-50 mM ethylene glycolbis(P-amino-ethyl ether)-N,N,N',N'-tetraacetic acid (EGTA)-1 mg of bovine serum albumin per ml. After 60 min of incubation at 37°C, acid-precipitable materiai vvas collected onto glass fiber filters by using a semiautomatic celi harvester. Bacterial celi extracts containing RT vvere diluted to vvithin the linear range of the assay, and activity vvas determined in the presence and absence of inhibitor. Purified HIV-1 RT heterodimer produced in E. coli also served as a control. Results are determined as inhibitor concentration to give 50% inhibition (ICso vvt), in nanomoles/liter.

[0078] For the double mutant assay (dm), A17 RT vvas employed in the assay. A17 RT is resistant to vanous aminopyridones, as described in Nunberg, J.H. £t aļ., J. Virai. 65, 4837 (1991). Results are measured as IC^ dm in nanomoles/liter.

CELL SPREAD ASSAY [0079] Inhibition of the spread of HIV in celi culture vvas measured according to Nunberg, J. H. et al., J. Virai. 65, 4887 (1991). In this assay, MT-4 T-lymphoid celis vvere infected vvith HIV-1 (wild-type, unless othervvise indicated) by using a predetermined inoculum, and cultures vvere incubated for 24h. At this time, <1% of the celis were positive by indirect immunofluorescence. Celis vvere then extensively vvashed and distributed into 96-vvell culture dishes. Serial tvvofold dilutions of inhibitor vvere added to the vvells, and cultures were continued for 3 additional days. At 4 days postinfection, 100% of the celis in control cultures vvere infected. HIV-1 p24 accumulation vvas directly correlated with virus spread. The celi culture inhibitory concentration vvas defined as the inhibitor concentration in nanomoles/liter vvhich reduced the spread of infection by at least 95%, or CIC_g5.

SUMMARY OF RESULTS FOR COMPOUND 37.2

A, Reverse Transcriptase Assav and Celi Spread Assav:

WT_K103N* Y181C DM RT-2

IC50(gM) 0.002 0.030 0.008 0.085 80.8

CIC95(gM) <0.006(N=2) 0.100 <0.025 0.400 N.D.

B, Phannacological Data;

Rhesus: 1 mg kg*¹ i.v.l_1y2a210 min.

mg kg *¹ p.o. (methocel): C_max=4.4gM @ 2h

Protein Binding: 98.0% Normai Human Plasma (HPLC Method) * Mutants Κ103 N and Υ181C are dmg-resistant HTV-1 reverse transcriptases. DM is the double mutant, as discussed in the reverse transcriptase assay. RT-2 is the reverse transcriptase of HIV-2.

O.B.=0.55 (A.U.C.)

SVNERGISTIC EFFECTS

A. Preparation of HlV-infected MT-4 celi Suspension [0080] MT celis vvere infected at Day 0 at a concentration of 250,000 per ml with a 1:1000 dilution of HIV-1 strain lllb stock (final 125 pg p24/ml; sufficient to yield S 1% infected celis on day 1 and 25-100% on day 4). Celis vvere infected and grown in the following medium: RPM11640 (VVhittaker BioProducts), 10% inactivated fetal bovine serum, 4 mM glutamine (Gibco Labs) and 1:100 Penicillin-Streptomycin (Gibco Labs).

[0081] The mixture vvas incubated overnight at 37°C in 5% CO₂ atmosphere.

B. Treatment vvith Inhibitors [0082] A matrix of nanomolar range concentrations of the pairwise combinations (see Table S) vvas prepared. At Day 1, aliquots of 125 μΙ of inhibitors vvere added to equal volumes of HlV-infected MT-4 celis (50,000 per vvell) in a 96vvell microtitercell culture plate. Incubation vvas continued for 3 days at 37°C in 5% CO₂ atmosphere.

C. Measurement of Vīrus Soread [0083] Using a multichannel pipettor, the settled celis vvere resuspended and 125 μΙ harvested into a separate microtiter plate. The supematant was assayed for HIV p24 antigen.

[0084] The concentration of HIV p24 antigen vvas measured by an enzyme immunoassay, described as follovvs. Aliquots of p24 antigen to be measured vvere added to microvvells coated vvith a monodonal antibody specific for HIV core antigen. The microvvells vvere vvashed at this point, and at other appropriate steps that follovv. Biotinylated HlV-specific antibody vvas then added, follovved 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 Svnergv [0085] Pairvvise combinations of inhibitors (see Table 5) vvere 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 pairvvise combination of 726 and AZT vvas found to exhibit markedly enhanced inhibition of vīrus spread, in comparison to 726 alone or AZT alone, or in comparison to the sum of 726 inhibition and AZT inhibition.

[0086] This data vvas processed as follovvs:

fractional inhibitory concentration ratios (RC) vvere calculated according to Elion, et al. J, Biol. Chem., 208 477 (1954). The minimum sum of FICS, vvhich is the maximum synergy, vvas determined for various pairvvise combinations. Altematively, an average sum of the FICS is calculated, vvhich is the average synergy. See Table S. These results indicate substantial synergy in the inhibition of virus spread. The smaller the number, the greater the synergy.

TABLE S

Pairvvise Combinations*	Average Synergy
726 + ddl	0.81
726 +AZT	0.62
726 + 524	0.65
726 + 524 + AZT
524 is L-735,524. Other compounds are also defined in Table C above.

[0087] While the foregoing specification teaches the principles of the present invention, vvith examples provided for the purpose of illustration, it vvill be understood that the practice of the invention encompasses ali of the usual variations, adaptions, or modifications, as come vvithin the scope of the following claims and its eguivalents.

Claims (16)

1. Izgudrojuma formula

   1. Savienojuma ar formulu (II) kurā:

   X ir halogēna atoms;

   X^I ir trihalogēnmetilgrupa, pentahalogēnetilgrupa, C^alkilgrupa,

   C2-5alkinilgrupa, C3.5cikloalkilgrupa, fenilgrupa, naftilgrupa, tetrahidronaftilgrupa, bifenililgrupa, fenetilgrupa, antrilgrupa, acenaftilgrupa;

   Z ir 0 vai S;

   R ir:

   (a) Ci^alkilgrupa, kas neobligāti aizvietota ar A, kur A ir:

   halogēna atoms, C^cikloalkilgrupa, ciāngrupa, hidroksilgrupa, Ci^alkoksigrupa, C₂^alkinil-Ci^alkoksigrupa, fenoksigrupa, naftiloksigrupa, tetrahidronaftiloksigrupa, bifenililoksigrupa, fenetoksigrupa, antroksigrupa, acenaftoksigrupa, C^alkilkarbonilgrupa, nitrogrupa, di(Ci.₂aIkil)aminogrupa, Ci^alkilamino-Cļ^alkilgrupa, heterocikliska grupa, feniltiogrupa, naftiltiogrupa, tetrahidronaftiltiogrupa, bifenililtiogrupa, fenetiltiogrupa, antriltiogrupa, acenaftiltiogrupa, kur heterocikliskā grupa ir stabila 5-7 locekļu monocikliska vai stabila 811 locekļu heterocikliskā grupa, kas ir nepiesātināta, daļēji nepiesātināta vai piesātināta un satur oglekļa atomus un 1-4 heteroatomus no rindas: N, 0 un S, pie kam slāpekļa un sēra heteroatomi ir neobligāti oksidēti, tai skaitā arī jebkura bicikliska heterocikliska grupa, kurā kāda no augstāk minētajām heterocikliskajām grupām ir kondensēta ar benzola gredzenu;

   (b) C₂^alkenilgrupa, neobligāti aizvietota ar:

   (i) A vai (ii) fenilgrupu, naftilgrupu, tetrahidronaftilgrupu, bifenililgrupu, fenetilgrupu, antril vai acenaftilgrupu, kas neobligāti aizvietotas ar A;

   (c) C2-5alkinilgrupa, neobligāti aizvietota ar:

   (i) A vai (ii) fenilgrupu, naftilgrupu, tetrahidronaftilgrupu, bifenililgrupu, fenetilgrupu, antril vai acenaftilgrupu, kas neobligāti aizvietotas ar A;

   (d) C^cikloalkilgrupa, neobligāti aizvietota ar:

   (i) A vai (ii) fenilgrupu, naftilgrupu, tetrahidronaftilgrupu, bifenililgrupu, fenetilgrupu, antril vai acenaftilgrupu, kas neobligāti aizvietotas ar A, vai tā farmaceitiski pieņemamās sāls pielietojums tāda medikamenta ražošanai, kas inhibē HIV apgriezto transkriptāzi, kas izmantojams HIV infekcijas profilaksei, HIV infekcijas ārstēšanai, AIDS vai ar AIDS saistītas saslimšanas ārstēšanai
2. 2. Pielietojums pēc 1. punkta, kurā savienojums ar formulu (II) ir: (-)-6-hlor-4-ciklopropiletinil-4-trifluormetil-1,4-dihidro-2H-3,1-benzoksazin-2ons, (-)-6-hlor-4-feniletinil-4-trifluormetil-1,4-dihidro-2H-3,1-benzoksazin-2-ons, (+/-)-6-hlor-4-(2-ciānfeniletinil)-4-trifluormetil-1,4-dihidro-2H-3,1 -benzoksazin2-ons, (+/-)-4-( 1-hlor-1,1-difluormetil)-4-(2-feniletinil)-6-hlor-1,4-dihidro-2H-3,1benzoksazin-2-ons, (+/-)-4-[2-(dimetilaminometil)etinil]-4-trifiuormetil-6-hlor-1,4-dihidro-2H-3,1 benzoksazin-2-ons, vai tā farmaceitiski pieņemama sāls.
3. 3. Savienojums ar formulu II pēc 1. vai 2. punkta, ar noteikumu, ka X¹ un R abi nav neaizvietota CMalkilgrupa, pielietojumam humānā medicīnā.
4. 4. Farmaceitiskā kompozīcija, kas satur savienojumu ar formulu (II) pēc 1. vai 2. punkta, ar noteikumu, ka X¹ un R abi nav neaizvietota Ci^alkilgrupa.
5. 5. Savienojums ar formulu (I) t

   X kurā:

   X ir halogēna atoms;

   X^I ir trihalogēnmetilgrupa, pentahalogēnetilgrupa;

   Z irO;

   R ir (a) C^alkilgrupa, kas neobligāti aizvietota ar A, kur A ir:

   halogēna atoms, C^cikloalkilgrupa, ciāngrupa, hidroksilgrupa, CMalkoksigrupa, C2^alkinil-Ci^alkoksigrupa, fenoksigrupa, naftiloksigrupa, tetrahidronaftiloksigrupa, bifeniiiloksigrupa, fenetoksigrupa, antroksigrupa, acenaftoksigrupa, Ci^alkilkarbonilgrupa, nitrogrupa, di(Ci.₂alkil)aminogrupa, C^alkilamino-Ci^alkilgrupa, heterocikliska grupa, feniltiogrupa, naftiltiogrupa, tetrahidronaftiltiogrupa, bifenililtiogrupa, fenetiltiogrupa, antriltiogrupa, acenaftiltiogrupa, kur heterocikiiskā grupa ir stabila 5-7 locekļu monocikliska vai satbila 811 locekļu heterocikiiskā grupa, kas ir nepiesātināta, daļēji nepiesātināta vai piesātināta un satur oglekļa atomus un 1-4 heteroatomus no rindas: N, O un S, pie kam slāpekļa un sēra heteroatomi ir neobligāti oksidēti, tai skaitā ari jebkura bicikliska heterocikliska grupa, kurā kāda no augstāk minētajām heterocikliskajām grupām ir kondensēta ar benzola gredzenu;

   (b) C₂^alkenilgrupa, neobligāti aizvietota ar (i) A vai (ii) fenilgrupu, naftilgrupu, tetrahidronaftilgrupu, bifenililgrupu, fenetilgrupu, antril vai acenaftilgrupu, kas neobligāti aizvietotas ar A;

   (c) C₂.5alkinilgrupa, neobligāti aizvietota ar:

   (i) A vai (ii) fenilgrupu, naftilgrupu, tetrahidronaftilgrupu, bifenililgrupu, fenetilgrupu, antril vai acenaftilgrupu, kas neobligāti aizvietotas ar A;

   (d) C^cikloalkilgrupa, neobligāti aizvietota ar:

   (i) A vai (ii) fenilgrupu, naftilgrupu, tetrahidronaftilgrupu, bifenililgrupu, fenetilgrupu, antril vai acenaftilgrupu, kas neobligāti aizvietotas ar A, vai tā farmaceitiski pieņemama sāls.
6. 6. Savienojums, proti:

   (-)-6-hlor-4-ciklopropiletinil-4-trifluormetil-1,4-dihidro-2H-3,1-benzoksazin-2ons, (-)-6-hlor-4-feniletinil-4-trifluormetil-1,4-dihidro-2H-3,1-benzoksazin-2-ons, (+/-)-6-hlor-4-(2-ciānfeniletinil)-4-trifluormetil-1,4-dihidro-2H-3,1-benzoksazin2-ons, (+/-)-4-(1 -hlor-1,1-difluormetil)-4-(2-feniietiniI)-6-hlor-1,4-dihidro-2H-3,1 benzoksazin-2-ons, (+/-)-4-[2-(dimetilaminometil)etinil]-4-trifluormetil-6-hlor-1,4-dihidro-2H-3,1benzoksazin-2-ons, vai tā farmaceitiski pieņemama sāls.
7. 7. Kombinācija, kas sastāv no savienojuma ar formulu (I) pēc 5. punkta vai formulu (II) pēc 1. vai 2. punkta vai tā farmaceitiski pieņemamās sāls un nukleozTda analoga ar bioloģisko aktivitāti pret HIV apgriezto transkriptāzi.
8. 8. AIDS antivirālo savienojumu kombinācija, proti:

   neobligāti viens vai vairāki HIV inhibitori no rindas:

   AZT, ddl un ddC.
9. 9. AIDS antivirālo savienojumu kombinācija, proti:

   un viens vai vairāki HIV inhibitori no rindas:

   AZT, ddl un ddC.
10. 10. Paņēmiens (-)-6-hlor-4-ciklopropiletinil-4-trifluormetii-1,4-dihidro2H-3,1-benzoksazin-2-ona (L-743,726) iegūšanai, kas ietver šādus posmus:

    (a) pietiekoša daudzuma (+/-)-6-hlor-4-ciklopropiletinil-4-trifluormetil1,4-dihidro-2H-3,1-benzoksazin-2-ona (L-741,211) nodrošināšana;

    (b) posmā (a) iegūtā savienojuma savienošana ar optisko izomēru atdalīšanas aģentu;

    (c) iegūto diastereomēru atdalīšana;

    (d) atdalīšanas aģenta aizvākšana vai pārveidošana, lai iegūtu vēlamo produktu.
11. 11. Paņēmiens pēc 10. punkta, kur optisko izomēru atdalīšanas aģents ir (-)-kamfanilhlorīds.
12. 12. (-)-6-Hlor-4-ciklopropiletinil-4-trifluormetil-1,4-dihidro-2H-3,1-benzoksazin-2-ons vai tā farmaceitiski pieņemama sāls.
13. 13. Farmaceitiskā kompozīcija, kas satur savienojumu pēc 12. punkta vai tā farmaceitiski pieņemamo sāli un farmaceitiski pieņemamu pildvielu.
14. 14. Savienojums pēc 12. punkta vai tā farmaceitiski pieņemama sāls pielietojumam ārstniecības paņēmienā.
15. 15. Savienojuma pēc 12. punkta vai tā farmaceitiski pieņemamas sāls pielietojums tāda medikamenta ražošanai, kas inhibē HIV apgriezto transkriptāzi, kas izmantojams HIV infekcijas profilaksei, HIV infekcijas ārstēšanai, AIDS vai ar AIDS saistītas saslimšanas ārstēšanai
16. 16. Savienojuma pēc 12. punkta vai tā farmaceitiski pieņemamas sāls kombinācija ar nukleozīda analogu ar bioloģisko aktivitāti pret HIV apgriezto transkriptāzi.