AP189A

AP189A - 5,11 Dihydro-6H-dipyrido (3,2-b:2, 3 -e)(1,4)-diazepin-6-ones and thions and their use for the treatment of aids.

Info

Publication number: AP189A
Application number: APAP/P/1990/000188A
Authority: AP
Inventors: Schmidt Gunther Dr; Engel Wolfhard Dr; Trummlitz Gunter Dr; Eberlein Wolfgang Dr; Hargrave Karl D Dr
Original assignee: Thomae Dr Karl GmbH; Boehringer Ingelheim Pharma GmbH Co KG; Boehringer Ingelheim Pharmaceuticals Inc
Current assignee: Thomae Dr Karl GmbH; Boehringer Ingelheim Pharma GmbH Co KG; Boehringer Ingelheim Pharmaceuticals Inc
Priority date: 1989-06-28
Filing date: 1990-06-28
Publication date: 1992-05-04
Anticipated expiration: 2010-06-28
Also published as: JPH03115283A; HUT55017A; HU206504B; KR910000727A; OA09741A; ZA904991B; CA2019812A1; NO902851L; ES2063202T3; IE65247B1; IL94883A; DE69007891D1; IE902322L; EP0410148A1; CA2019812C; FI92828B; HU904021D0; AU620724B2; JP2911967B2; ATE103918T1

Abstract

Disclosed are novel 5,11-dihydro-6h-dipyrido[3,2-b; 2',3'-e][1,4]diazepin-6-ones and thiones. These are useful in the prevention or treatment of aids.

Description

The invention relates co novel 5,ll-dihydro-6H-dipyrido(3,2-b:

' , 3 '-e if1 jdiazspin-6-or.es and -chiones and pharmaceutically accepcaole acid addition sales thereof, methods for preparing these compounds, the use of these compounds in the prevention or treatment of AIDS, and to pharmaceutical compositions containing these compounds.

3ackgrour.d of the Invention

The human disease, Acquired Immune Deficiency Syndrome (AIDS), is caused by the Human Immunodeficiency Virus (HIV), particularly the strain known as HIV-1.

Like ocher viruses, HIV-I cannot replicate without commandeering the biosynthetic apparatus of the host cell it infects. It causes this apparatus to produce the structural proteins which make up the viraL progeny. These proteins ara coded for by the genecic material contained within the infecting virus particle, or virion. Being a recrovirus, however, the genetic material of HIV Is RNA, not DNA as in the host cell's genome. Accordingly, the viral RNA must first be converted into DNA, and then integrated inco the hose cell's genome, in order for the host cell to oroduce the required viral proceins.

AP 0 0 0 1 8 9

BAO ORIGINAL

The conversion of she ?NA so DMA is accomplished through she use of she enzyme reverse transcriptase (RT), which is included within she infecting virion along wish she RNA. Reverse cranscriocase has three enzymatic functions; it acts as an RNA-dependent DNA polymerase, as a ribonuclease, and as a DNA-dependenc DNA polymerase. Acting first as an RNA-dependent DNA polymerase, RT makes a single-stranded DNA copy of the viral RNA. Next, acting as a ribonuclease, RT frees the DNA just produced from the original viral RNA and then destroys the original RNA. Finally, again acting as a DNA-dependenc DNA polymerase, RT makes a second, complementary DNA strand, using the first DNA strand as a template. The two strands form double-stranded DNA, which is integrated into the host cell’s genome by another enzyme called an integrase.

Compounds which inhibit the enzymatic functions of HIV-1 reverse transcriptase will inhibit replication of HIV-1 in infected cells. Such compounds are useful in the prevention or treatment of HIV-1 infection in human subjects.

Description of the Invention

In one of its composition of matter aspects, the invention comprises 5,11 -dihvdro-6H-dipvrido ι 3,2 -b: 2’ , 3 ’ -e; [ 1.4!diazepin-6-ones and -thiones of the formula

wherein,

Z is oxygen or sulfur;

rA is hvdrogen, alkyl or fluoroalkyl of 1 co 5 carbon acorns. alkenyl or alkvnyl of 3 co 5 carbon acorns, 2-halo-propen-l-yl, arylmechyl (wherein che aryl moiety is phenyl, chienyl or furanyl which is either unsubscituted or subscicuced by methyl, mechoxy or halogen), alkanoyl of 2 co 3 carbon acorns, or alkoxyalkyl or alkylchioalkyl of 2 co i carbon acorns;

hydrogen*

R.2 is/alkyl or fluoroalkyl of 1 co 5 carbon acorns, alkenyl or alkynyl of 2 co 5 carbon acorns, alkoxyalkyl or alkylchioalkyl of 2 co 4 carbon atoms, alkanoyl of 2 co 4 carbon acorns, hydroxyalkyl of 2 co 5 carbon acorns, arylmechyl· (wherein che aryl moiecy is phenyl, chienyl or furanyl, which is eicher unsubscicuced or subscicuced by alkyl or alkoxy of 1 co 3 carbon acorns, hydroxyl or halogen), phenyl (which is eicher unsubscicuced or subscicuced by alkyl or alkoxy of 1 co 3 carbon acorns, hydroxy or halogen) or alkoxycarbonylmechyl wherein che alkoxy moiecy concains 1 co carbon acoms; and i a

R chrough R are each hydrogen; or, one of r3 chrough R® is alkyl of 1 co 4 carbon acoms, alkoxy or alkylthio of 1 co 4 carbon acoms, alkoxycarbonyl of 2 Co 4 carbon acoms, hydroxyalkyl of 1 Co 4 carbon acoms, alkanoyl of 2 co 4 carbon acoms, alkanoyloxy of 2 co 4 carbon acoms, alkanoylamino of 1 to 4 carbon atoms, arainoalkyl of 1 to 4 carbon acoms, alkoxycarbonylalkyl wherein che alkoxy and alkyl moiecies each contain 1 co 2 carbon acoms, carboxyalkyl of 2 co ώ carbon acoms, mono- or di-alkylamino wherein each alkyl moiecy concains 1 co 2 carbon acoms, cyano, nitro, hydroxyl, carboxyl, amino, mono- or

AP 0 0 0 1 8 9

BAD ORIGINAL di-alkylaminoalkyl vherein each alkyl moiety contains 1 co 2 carbon acorns, acido or halogen, with che ocher five substituents being hydrogen;

or

R.3₁ 2!*, and R? _{are eac}h independently hydrogen or alkyl of 1 co 3 carbon acorns, with che proviso chac ac lease one of these subscicuencs is hydrogen, or one of R^J , R and R^J is butyl with che remaining two subscicuencs being hydrogen; and, r6 , R~ , and R³ are each independently hydrogen or alkyl of 1 co 3 carbon atoms, wich che proviso chac ac least one of these subscicuencs is hydrogen, or one of R³ , R⁷ and R³ is butyl wich che remaining two subscicuencs being hydrogen^ with the proviso that when Z is oxygen and R and R are the same or different and are hydrogen or straight chained or branched alkyl of 1 to 5 3 8 carbon atoms at least one of R^J through R is other than hydrogen.

A subgeneric aspect of che invention comprises compounds of formula I, wherein,

Z is oxygen or sulfur:

rJ· is hydrogen, alkyl or fluoroalkyl of 1 co 5 carbon acorns, crihalomethyl, alkenyl or alkynyl of 2 to 4 carbon atoms, 2-halo-propen-l-yl, or alkoxyalkyl or alkylthioalkyl of 2 to 3 carbon atoms;

R.Z is alkyl or fluoroalkyl of 1 co 4 carbon atoms, alkenyl or alkynyl of 2 co 4 carbon atoms, alkoxyalkyl or alkylthioalkyl of 2 co 4 carbon acorns, alkanoyl of 2 co 3 carbon acorns, hydroxyalkyl of 2 to 4 carbon atoms, arylmethyl (wherein che aryl moiecy is phenyl or thienyl, which is either unsubscituted or substituted by mechyl, methoxy, hydroxyl or halogen), phenyl (which is either unsubscicuted or subscicuced wich_s

BAD ORIGINAL mechvl., mechoxy, hydroxyl or halogen) or alkoxycarbonyImethyl wherein che alkoxy moiecy contains 1 co 5 carbon acorns ;

r\ g/⁴, and R³ are each independently hydrogen or methyl, with the proviso chac ac lease one of chese substituents is hydrogen, or R^ £_sethyl, propyl or butyl with the remaining two substituents being hydrogen; and,

R^, R? , and R^ are each independently hydrogen or methyl, with the proviso that ac least one of chese substituents is hydrogen, or R^ is ethyl, propyl or butyl with the remaining two substituents being hydrogen.

A particular subgeneric aspect of the invention comprises compounds of formula I wherein.

Z is oxygen or sulfur;

R^· is hydrogen, alkyl or fluoroalkyl of 1 to 4 carbon atoms or allyl; r2 is alkyl or fluoroalkyl of 1 to 4 carbon atoms, allyl or benzyl; and

R through R are each hydrogen.

The compounds of Formula I can be prepared by known methods or obvious modifications thereof. Methods A. 3. C, D, and E, described below, are illustrative of the methods for preparing the compounds.

Method A

Compounds of the general formula Ia y

AP 0 0 0 1 8 9

BAD ORIGINAL

wherein sA and R^ chrough R^ are defined as above and R² has the same definitions as R² with the exception of hydrogen, can be obtained by cyclizing carboxylic acid amides of general formula II

Ha 1 HN

2' wherein rA, rA Chrough R® and R² have the same definitions sec forth with respect to Formula la and Hal represents fluorine, chlorine, bromine or iodine. Cyclisation is preferably carried out by converting the compounds of general formula II into their alkaline metal salts and subsequent condensation ac temperatures between O’C and the boiling point of the reaccion mixture.

If, in the starting compounds of general formula II. R^ is different from hydrogen, metallacion requires ac least 1 mole of the metallacing agent. If on the other hand, rA Is hydrogen, at least 2 moles of this agent'?musc be used. For metallacion. lithium, sodium and potassium hydrides, lithium alkvls, such as n-bucyl lithium, are preferably used.

BAD ORK3TNAL

The reaction is usually carried out in inerc solvents, e.g. in tecrahydrofuran. 1,i-dioxane. glycoldimethyl ether.

die thy leneglycoldi.ne thyl ether, triethyleneglycoldimechyl ether, dimethylformamide. benzene or anisole. Cyclisation may also be effected by heating carboxylic acid amides of general formula II in dipolar aprotic solvents, preferably in sulfolane or dimechylsulfone. Catalytic quantities of strong acids, e.g. sulfuric acid, hydrochloric acid, * hydrobromic acid, phosphoric acid, polyphosphoric acid, mechanesulfonic acid or p-coluenesulfonic acid, have proved to be of use. The necessary reaction temperature is usually between 110 and 220*C, the preferred range of temperature being between 130 and 170’C.

Method B

Compounds of general formula lb

lb

AP 0 0 0 1 8 9 wherein R^hydro lytic and R^ through cleavage of the

Q

R are defined as above, can.be prepared by arylmethyl group in compounds of general

BAD ORIGINAL wherein R* 3r.d R' through R^ are defined as mentioned above and Ar can be, for examrle. a phen.vl or i-methoxyphenyl group . Hydrolysis is affected bv moderate ro strong acids or Lewis-acids ar temoenatures between -20 and -150°0. Such acids can be, for example, sulfuric acid, mecnanesulfonic acid, rrifluoroacecic acid, crifluoromerhanesulfonic acid, phosphoric or polyphosphoric acid. LRien using phosphoric or polyphosphoric acid, rhe addition of solvents such as benzene, toluene, phenol, anisole or veracrole has proved co be of advantage.

such as aluminum chloride or bromide are used to ylmechvL group, solvents such as aromatic hydrocarbons, luene, anisole, or mixtures thereof with dichloromethane

If Lewis acids, eliminate rhe ar

e.g. benzene, to are suitable.

It will be obvious to those skilled in the art that Method 3 is not preferred in those cases wherein any or R·*· and R through R° are readily hydrolyzable substituents, for example, wherein R^· is alkanoyl or any of R^ through r3 are alkanoylamino or alkoxycarbonyl. In cases wherein R^· fl is alkanoyl or any of R^J through R are alkoxycarbonyL, for example, it is preferable to utilize method A described above; when R^· is hydrogen two equivalents of base must be used. In cases wherein any of R^ through

3.3 are alkanoylamino, for example, it is preferable to carry out the hydrolysis (and subsequent acylation) on the corresponding nitro derivative, and then reduce the nitro moiety to the amine, followed by acylation to yield the desired product.

Method C compound of general formula lc ·*ν

BAD ORIGINAL ο

wherein κΛ has the same definitions as rA with the exception of hydrogen and through R^ _are defined as above, may be obtained by converting a 5 , ll-dihydro-0H-dipyrido[3,2-b:2'.3'-e](l,4jdiazepin-6-one of the formula

IV

(IV) wherein 3? through R® are defined as above, into the corresponding

5-alkali or alkaline earth metal compound and subsequently reacting the alkali metal compound with a compound of the formula V

AP 0 0 0 1 8 9 »

wherein R⁴· has the same meanings as in formula Ic and X is the radical

I of a reactive ester, a halogen atom, the group OSO2OR . the methanesulfonyloxy or echanesulfonyloxy group or an aromatic sulfonyloxy group. Instead of converting the compound of the general formula IV into its corresponding alkali metal salt in the first step, the alkylation of a compound of formula IV may also be performed by reaction with a compound of formula V in the presence of amines, such as triethylamine, dia£abicycloundecene or 4-(dimechylamino)pyridine , or of alkali

BAD ORIGINAL carbonates or bicarbonates. such as sodium and potassium carbonate or sodium bicarbonate.

The conversion of a compound of general formula IV into the corresponding alkali metal or alkaline earth metal compound may be effected by reacting a compound of formula IV with an alkali metal or alkaline earth metal hydroxide, such as lithium hydroxide, barium hydroxide, sodium hydroxide or potassium hydroxide, with an alkali metal alco'nolate, such as sodium methanolate or potassium tert-butoxide, with an alkali metal amide, such as sodium amide or potassium amide, or with an alkali metal hydride such as sodium hydride or potassium hydride. The reaction is preferably carried ouc at elevated temperatures and in the presence of a suitable organic solvent. Inert organic solvents, such as tetrahydrofuran or glycoldimethyl ether are preferred if alkali metal hydrides are used as . the metallating agents, whereas, if an alkali or alkaline earth metal hydroxide is used, an aqueous mixture with an organic solvent, such as methanol or tetrahydrofuran, may also be employed. For conversion of the alkali or alkaline earth metal-substituted 5, IL-dihydro-6H-dipyrido (3,2-b:2',3'-eJ[1,4]diazepin-5-one thus obtained into a compound of general formula lc, the solution or suspension of the alkali or alkaline earth metal compound is reacted directly, i.e. without isolation, with a compound of formula V at -20*C or at elevated temperatures, up to the boiling point of the solvent or reaction medium, whichever is lower. The substitution cakes place almost exclusively at the nitrogen atom in the

5-position of the dihydrodipyridodiatepinone, even if R^fc in the starting material of formula IV is a hydrogen atom, provided that one equivalent of base and one equivalent of a compound of formula V are used.

bad ORIGINAL

It will be obvious to those skilled in the art that the presence of nucleophilic substituents in the compounds of formula Ic may require the use of an intermediate of formula Ic having substituents which are, ocher chan the 11-posicion nitrogen, not nucleophilic buc which can be derivacized to yield the required group. For example, amino or monoalkylamino substituents ac any or R througn R are preferably obtained by alkylating or acylating an incermediace of formula Ic having γ a a nitro group at any of R^J through R°, and subsequently reducing the nitro group, and alkylating, if appropriate, co yield the final product.

Method D

(Id)

AP 0 0 0 1 8 9 * 7 wherein R has the meanings of R with the exception of 'alkanoyl, hydroxyalkyl or alkoxycarbonylmethyl, and R^· and R^ through R® represent the groups mentioned above, can be obtained by converting a ,11-dihydro-6H-dipyrido[3,2-b:2',3’-3ji1,4]diazepin-6-one of general formula lb into the corresponding metal salt of general formula Via or in the case of R^· in the compound of formula lb being hydrogen - into a compound of formula VIb

wherein M represents an alkali metal, such as lithium, sodium, potassium, rubidium or cesium, or M represents the group MgHal+, wherein Hal is a chlorine, bromine or iodine atom, and subsequently alkylating with a compound of general formula VII

VII wherein ,2 and X are as hereinbefore defined.

The conversion of a compound of general formula lb into the corresponding alkali metal compound of formuLae Vid and VIb may be effected by reacting a compound of formula lb with a Lithium alkyl (e.g. n-butyl lithium, or c-butyl lithium) optionally in the presence of tetramethylethylenediamine. a lithium dialkylamide, (e.g. lithium rad original diisoorooviamide, lithium dicyclohexylamide and lithium isoprooyicyclohexyiamide), a lithium aryl (e.g. phenyl lithium), an aikali metal hvdroxide (e.g. lithium, sodium or potassium hydroxide), an alkali mecal hydride (e.g. sodium or potassium hydride), an aikali metal amide (e.g. sodium or potassium amides) or a Grignard reagent (e.g. methyl magnesium iodide, ethyl magnesium bromide or phenyl magnesium bromide). One equivalent of base is required for the formation of compounds of formula Via, whereas two equivalents of base are required for the formation of compounds of formula VIb. The metallacion is conveniently carried out in an inert organic solvent at temperatures of between -78*C and the boiling point of the reaction mixture in question. If a Lithium alkyl, lithium aryl, 1 ichium dialkylamide or Grignard reagent is used for the metallacion, the preferred solvents are ethers such as tetrahydrofuran, diethyl ether or dioxane, optionally in a mixture with aliphatic or aromatic hydrocarbons, such as hexane or benzene, and the operation may be carried out at temperatures of becween -20 and +80*C. When metallacion is effected with an alkali mecal hydride or alkali metal amide, in addition co the solvents mentioned hereinbefore it is also possible to use xylene, toluene, acetonitrile, dimechylformamide and dimechylsulfoxide, while if an alkali metal hydroxide is used ic is also possible to use alcohols such as ethanol, methanol and aliphatic ketones such as acetone, as well as mixtures of these solvents wich water.

For conversion of the alkali mecal salt thus obtained inco a compound of formula Id, the solution or suspension of the alkali metal compound is reacted directly, i.e. without isolation of the reaction product, with a compound of formula VII at -20’C or at elevated temperatures, preferably

AP 0 0 0 1 8 ac the boiling point of the solvent or suspension medium or at the boiling point of the compound VII, whichever is lower.

Ic will be obvious to those skilled in the art chat the presence of nucleophilic substituents in the compounds of formula Id may require the use of an intermediate of formula Id having subscituents which are, other than the 11-position nitrogen, not nucleophilic but which can be derivatited to yield the required group. For example, amino or *) a monoalkylamino substituents at any or R^J through R are preferably obtained by alkylating or acylating an intermediate of formula Ic having a nitro group at any or R through R , and subsequently reducing the nitro group, and alkylating, if appropriate, to yield the final product.

The carboxylic acid amides of general formula II used as starting materials are obtained, for example, by amination of 2-chloro-nicocinic acid amides of general formula VIII

(VIII)

Hal Cl wherein R^· through R^ and Hal are as hereinbefore defined amines of general formula IX

HnM-R²' with primary (IX)

L4

BAD ORIGINAL wherein R^' is as hereinbefore defined. The reaction can also be carried out in the presence of inorganic or organic auxiliary bases, such as triethylamine. N, .V-dime thy Laniline , or sodium or potassium carbonate.

The reaction- can be carried out without using a solvent; it is of some advantage, however, to use inert organic solvents at temperatures of between 0°G and 1;O°C, preferably at reflux temperature. Suitable inert solvents that can be used include an excess of the primary amine of general formula IX. open chain or cyclic ethers, such as tecrahydrofurari,

1,4-dioxane, glycoldimethyl ether, diethyleneglycoldimethyl ether; aromatic hydrocarbons, such as benzene, toluene, xylene, chlorobenzene or oyridine; alcohols such as methanol, ethanol, isopropanol; dipolar aprotic solvents such as dimethylformamide; 1,3-dimethyl-2imidazolidinone, l,3-diaethyl-tatrahydro-2(lH)- pyrimidinone and sulfolane. Starting materials of general formula VIII, wherein rA is different from hydrogen, can be prepared from 2-chloronicotinic acid amides of general formula X

AP 0 0 0 1 8 9 by reaction with alkylating agents of general formula V in the presence of proton acceptors, for example of amines, such as triethylamine, diazabicycloundecene, 4-(dimethylamino)pyridine, or alkali or alkaline earth metal hydroxides, such as sodium hydroxide, potassium hydroxide, calcium hydroxide, of alkali carbonates, or alkaline earth metal carbonates or hydrogencarbonates, such as sodium carbonate or potassium carbonate, or potassium hydrogen carbonate.

BAD ORIGINAL

2-Chloror.icocinic acid amides of general formula X can be obtained bv condensation of 2-chloror.icocinic acid chloride with 3-amino-2halopvridines , under well known reaction conditions.

All the ocher starting materials are known from the literature or may be purchased or may be obtained by procedures known from the literature.

Method £

In Method E, a compound of Formula I, wherein Z is sulfur, is obtained by reacting a compound of Formula I. wherein Z is oxygen, with a sulfurating agent, such as 2,4-bis (4-mechoxyphenyl) -1,3-dichia-2 ,-ΐϋρηοβρηβ tan.e - 2 , d-disulfide; bis(cricyclohexyltinj sulfide ; bis (tri-nbutylcin)sulfide ; bis (criphenyltin)sulfide; bis(crimechylsilyl)sulfide or phosphorous pentasulfide. The reaction is carried out in an inert organic solvent such as carbon disulfide, benzene or toluene, at room temperature or higher, preferably ac an elevated temperature up. to the boiling point of the reaction mixture, and preferably under anhydrous conditions. When using the above mentioned tin or silyl sulfides, ic is preferable to carry out the sulfurization reaction in the presence of a

Lewis acid such as boron trichloride.

It will be obvious to those skilled in the arc thac the presence of another carbonyl moiety in a compound of formula I, for exampLe, a compound wherein Z is oxygen and any of R^J through R is aixanoyl, will require that the ketone carbonyl be protected via known methods by a suitable protecting group prior to the sulfurization reaction; deprocection subsequent to the sulfurization reaction provides the desired compound. Similarly, in cases wherein R^ is, for example, alkanovl, it will be obvious thac Che suLfurization reaction should be

BAD ORIGINAL

9/038-l-Cl oean.

In hose

from nitro, for example. alkanoylaraino. the sulfurization reaction can be performed on che corresponding nitro derivative, followed by an appropriate (known) reduction and finally acylation co yield che desired produce.

Compounds of formula I may, if desired, be converted into their non-toxic, pharmaceutically acceptable acid addition salts by conventional methods: for example, by dissolving a compound of formula I in a suitable solvent and acidifying the solution with one or more molar equivalents of che desired acid. The invention also comprises such salts .

Examples of inorganic and organic acids which may form noncoxic, pharmaceutically acceptable acid addition salts with a compound of the formula I are the following: hydrochloric acid, hydvobromic acid, sulfuric acid, phosphoric acid, nitric acid, tartaric acid, citric acid, me thanesulfonic acid, and the like. Compounds of general formula I usually form acid addition salts with one molar equivalent of che acid.

The above described compounds of Formula I possess inhibitory activity against HIV-1 reverse transcriptase. When administered in suitable dosage forms, they are useful in the prevention or treatment of AIDS, ARC and related disorders associated with HIV infection. Another aspect of che invention, therefore, is a method for preventing or treating HIV-1 infection which comprises administering Co a human being, exposed to or

AP 0 0 0 1 8 9

9/O38-1-C1

BAD ORIGINAL infected by H'V-1, a orophylacricaily or therapeutically effective amount of a novel compound of Formula I, as described above.

The compounds of formula I may be administered in single or divided doses by the oral, parenteral or topical routes. A suitable oral dosage for a compound of formula I would be in Che range of about 10 to 500 mg per day. In parenteral formulations, a suitable dosage unit may contain from 1 to 50 mg of said compounds, whereas for topical administration, formulations containing 0.01 to 1% active ingredient are preferred. Ic should be understood, however, that the dosage administration from patient to patient will vary and the dosage for any particular patient will depend upon the clinician's judgement, who will use as criteria for fixing a proper dosage the size and condition of che patient as veil as the patient's response co the drug.

'Λβη the compounds of the present invention are to be adminiscerted by che oral route, they may be administered as medicaments in the form of pharmaceutical preparations which contain chem in association with a compatible pharmaceutical carrier material. Such carrier material can be an inert organic or inorganic carrier material suitable for oral administration. Examples of such carrier materials are water, gelatin, talc, starch, magnesium stearate, gum arable, vegetable oils, polyalkylene - glycols , petroleum jelly and che like.

The pharmaceutical preparations can be prepared in a conventional manner and finished dosage forms can be solid dosage forms, for example, tablets, dragees, capsules, and che like, or liquid dosage forms, for example solutions, suspensions, emulsions and che like. The pharmaceutical preparations may be subjected to conventional

BAD ORIGINAL

9/038-l-CL pharmaceutical operations such as sterilitacion. Further, che pharmaceutical preparations may contain conventional adjuvants such as preservacives, stabilizers, emulsifiers, flavor - improvers , wetting agents, buffers, salts for varying the osmotic pressure and the like. Solid carrier material which can be used include, for example, starch, lactose, mannitol, methyl cellulose, microcrystalline cellulose, talc, silica, dibasic calcium phosphate, and high molecular weighc polymers (such as polvechylene glycol).

For parenteral· use, a compound of formula I can be administered in an aqueous or non-aqueous solution, suspension or emulsion in a pharmaceutically acceptable oil or a mixture of liquids, which may contain bacteriostatic agents, antioxidants, preservatives, buffers or ocher solutes to render the solution isotonic with the blood, chickening, agencs, suspending agents or other pharmaceutically acceptable additives. Additives of this type include, for example, tartrate, citrate and acetate buffers, ethanol, propylene glycol, polyethylene glycol, complex formers (such as EDTA), antioxidants (such as sodium bisulfite, sodium metabisulfite, and ascorbic acid), high molecular weight polymers (such as liquid polyethylene oxides) for viscosity regulation and polyethylene derivatives of sorbitol anhydrides. Preservatives may also be added if necessary, such as benzoic acid, mechyl or propyl paraben, benzalkonium chloride and other quaternary ammonium compounds.

AP 0 0 0 1 8 9

The compounds of this invention may also be administered as solutions for nasal application and may contain in addition to the compounds of this invention suitable buffers, tonicity adjusters, microbial preservatives, antioxidants and viscosity-increasing agencs in an aqueous vehicle. Examples of agencs used co increase viscosity are polyvinyl alcohol.

9/038-1-Cl

BAD ORIGINAL cellulose derivatives. polyvinylpyrrolidone, polysorbates or glycerin. Microbial preservatives added may include benzalkonium chloride, thimerosal, chlorobutanol or phenylethyl alcohol.

Additionally, the compounds provided by che invention can be administered by suppository.

As staced before, che compounds provided by che invention inhibic che enzymatic activity of HIV-1 RT. Based upon testing of these compounds, as described below, it is known chac they inhibic che RNA-dependent DNA polymerase activity of HIV RT. 3ased upon ocher testing, noc described herein, it is believed chat they also inhibic che DNA-dependent DNA polymerase activity of HIV RT.

Utilizing che Reverse Transcripcase (RT) Assay described below, compounds can be tested for cheir ability to inhibic che RNA-dependent DNA polymerase activity of HIV RT. Certain specific compounds described in che Examples which appear below, were so tested.

REVERSE TRANSCRIPTASE (RT) ASSAY .Assay .neory;

Among the enzymes for which Human Immunodeficiency Virus (HTV-1) encodes is a reverse transcriptase (1), so-named because ic transcribes a DNA copy from an RNA template. This activity can be quantitatively measured in a cell-free enzyme assay which has been previously described (2), and is based upon the observation that reverse transcripcase is able to use a synthetic template [poly r(C) primed with oligo d(G)] to transcribe a

BAD ORIGINAL

9/038-1-Cl radio - labelled, acid-precipitable DNA strand utilizing ^H-dCT? as a substrate.

Materials:

a) Preparation of the enzyme

Reverse transcriptase enzyme from the LAV strain of Human Immunodeficiency Virus (HIV-1) (1) was isolated from the bacterial strain JM109 (3) expressing the DNA clone p3RTprtl+ (2) which is under the control of the lac promotor in the expression vector px3I21 (4). An overnight culture grown in 2ΧΪΤ medium (37*C, 225 rpra) (5) supplemented with ICO wg/mx ampicillin for positive selection is inoculated at a 1:40 dilution inco M9 medium supplemented with lOug/ml thiamine, 0.5% casamino acids, • .»». · and 50 ug/ral ampicillin (5). The culture is incubated (37*C, 225 rpm) until it reaches an QD54Q of 0.3-0.4. At that time the repressor inhibitor IPTG (isopropyl b-D-thiogalactopyranoside) is added to 0.5mM and incubated for 2 additional hours. Bacteria are pellected, resuspended in a 50mM Tris, 0.6mM EDTA, O.375M NaCl buffer and digested by the addition of lysozyme (lmg/ral) for 30 minutes on ice. The cells are lysed by the addition to 0.2% NP-40 and brought to 1M NaCl.

After removal of the insoluble debris by centrifugation, the protein is precipitated by the addition of 3 volumes of saturated aqueous ammonium sulfate. The enzyme is pelleted, resuspended in RT buffer (50mM Tris pH 7.5, ImM EDTA, 5mM DTT, 0.1% NP-40, 0.1M NaCl, and 50% glycerol), and stored at -70*C for further use.

AP 0 0 0 1 8 9

b) Composition of 2X concentrated stock reaction mixture

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9/038-1-C1

Stock ’eater.t

IX Mix Concentration

1M	Tris pH 7-.4	lOOmM
1M	Dithiochrietol	-OmM
LM	NaCl	12 OmM
1%	Nonidec ?-40	0.1%
LM	MgCl	4mM

[poly r(C) /oiigo d(C)](5:l) 2ug/ml ³H-dGT? (SluM) 0.6uM

Assay Procedure:

The 2X concentrated stock reaction mixture is aiiquoted and stored at -20*C. The mixture is stable and chawed for use in each assay. This enzyme assay has been adapted to a 96 well microtiter plate system, and has been previously described (6). Tris buffer (50 mM, pH 7.4), vehicle, (solvent diluted to match “he compound dilution), or compounds in vehicle are dispensed into 96-well microciter places (10ul/well; 3 wells/ compound). The HIV RT enzyme is thawed, diluted in 50mM Tris pH 7.4 so that fifteen ul of diluted enzyme contain 0.001 Unit (one unit is that amount of enzyme to transform 1 micromole of substrate per minute at 25*C), and fifteen ul are dispensed per well. Twenty ul of O.12-O.5M EDTA are added to the first three wells of the microciter plate. EDTA chelates the Mg^4-* present and prevents reverse transcription. This group senes as background polymerization which is subtracted from all ocher groups. Twenty-five ul of the 2X reaction mixture are added to all wells and the assay is allowed to incubate at room temperature for 60 minutes. The assay is terminated by precipitating the DNA in each well with 50ul of 10% trichioracecic acid (TCA) in 1% sodium pyrophosphate. The microtiter plate is incubated for 15 minutes ac 4*C and the precipitate

9/038-L-CL

BAD ORIGINAL is fixed onto =>20 glass fiber paper (Schleicher & Schuell) using a Skacron semi-automatic harvester. The filters are Chen washed with additional 5% TCA containing 1% sodium pyrophosphate, rinsed with 70¾ aqueous ethanol, dried, and transferred to scintillation vials (6). Each vial receives 2 mis of scintillation cocktail and is counted in a Beckman beta counter.

Calculations for percent inhibition are as follows:

% inh ib i c i on - C?M Mean Test Value - C?M Mean Control Value X100

C?M Mean Control Value

Re ferences:

1. Benn, S., et al., SCIENCE 230:949, 1985

2. Farmerie, W.G. et. al.. SCIENCE 236:305, 1987

3. Yanisch-Perron, C., Viera, J., and Messing, J.. GENE 33:103, 1985

4. International Biotechnologies, Inc., New Haven, CT 06535

5. Maniacis. T, Fritsch, E.F.. and J. Sambrook, eds. MOLECULAR

CLONING: A LABORATORY MANUAL, Cold Spring Harbor Laboratory,

1982

6. Spira, T. . et. al. J. Clinical Microbiology, 25:97, L987.

Ln order to confirm that compounds which are active in .the RT Assay also have the ability to inhibit HIV replication in a living system, compounds according to the invention were also tested in the human T-Cell Culture Assay described below.

HYMAN T CELL CULTURE ASSAY

MJ 0 0 0 1 8 9

Assay Theory: Formation of syncytia is a feature of in vitro cultures of CD4+ T-cells infected vith HIV-1. In this assay, T-cells are created with a putative replication inhibiting compound and then infected with

9/038-1-C1

BAD ORIGINAL

HIV-L. After incubation the culture is checked for the formation of svncvtia. The absence or reduction is the number of syncytia is used as a measure of the test compound's ability to inhibit HIV replication.

Assay Mechod: The target cells, designated C3160. are a subclone of human lymphoma cells of 7-cell origin and are established at an initial density of ixlO¹ per 100 ul in RPMI 1640 (+ 10% fatal bovine serum) culture medium in 96 well flat bottom plates. A selected amount of test compound, dissolved in DMSO is included. After 24 hours, 50-100 TCIDcq._s(the dose that results in induced effect in 50% of test cultures) of the

HTLV-”’3 strain of HIV-1 (2) are innoculated into each culture. Control cultures receive compound or virus only. Four days after virus challenge, cultures are visually examined for the frequency and distribution of virus - induced giant cell syncytia. The percent inhibition by the test compound is determined by comparison with control values. Confirmation of the presence or absence of virus replication is accomplished by harvesting the cell free culture fluids from all experimental groups to determine the presence or absence of infectious progeny through the induction of syncytia formation in secondary human T-cell cultures after 3 days.

References :

(1) M. Somasundaran and H.L. Robinson, Science 242. 1554 (1998) (2) G.M. Shaw, R.H. Hahn, S.K. Arya, J.S. Groopman, R.C. Gallo and F. Wong-Scaal, Science 225, 1165 (1984)

In order to assess the specificity of Che enzyme inhibitory activity of Che compounds provided by the invention, a few were tested, using known per se assay methods, for their ability to inhibit Feline Leukemia Virus BAD ORIGINAL

9/O38-1-C1 zeri'-'ed reverse trans crip case ar.d Calf Thymus-derived CA aloha-co?~erasa . More of the compounds so tasted was observed to possess any inhibitory activity against these enzymes. These results indicata that the enzyme inhibitory activity of the compounds provided bv the invention is directed rather specifically against HIV 2.T.

In order to roughly assess the cytotoxicity of the compounds provided by the invention, several such compounds were tasted in the MTT Cellular Cvtotoxic icy Assay described below.

Compounds having a relatively high EC^ are preferred.

HTT ASSAY FOR CELLULAR CYTOTOXICITY

Assay Theory:

The HTT ι 3 - (4, 5-dimechyichiazol-2yl)-2.5 diphenyl catrazolium bromide) assay is based on cleavage of tatrazoLium bromide by mecabolically active ceils, resulting in a highly quantitative blue color. This assay has been previously described (1) but has' been oocimized for che purposes of the testing reported herein.

AP 0 0 0 1 8 9

Assay hechod:

The H9 cell line (2), an established human lymphoma suspension ceil line grown in EPhI 1540 supplemented with 10% fecal bovine serum is used as the target ceil line in the assay. Ceils (lOOul) ara placad in microcasc place wells ac a concentration of 10° cells per ml in Che presence of varying concentrations of inhibitor. The ceils are incubated at 37’C in a humidified CO₂ incubator. Five days later, 20ul of MTT (5mg/ral in EPMI 1540. sonicated, 0.2 micron filtered, and stored ac 4’C) is added to each well. After 4 hours additional incubation ac 37*C, 50ul of Triton-X is

9/O38-1-C1

-—

BAD ORIGINAL added io each -e cr-’sn’.s. Ab sol iaaediacely read

1 and chor	oughly mined co aid Che	s olub i1i c a	cion of che
ce echanol	(5ul) is incuba cad for	10 ainuces	ac 50*0 and
on a place	reader (Dynacach) ac a	•-ave Ien.gch	of 57Cna.

Daca from “his assay are used co generace a nonlinear regression analysis vhich yields an SCsq.

References:

1. Mosaar.n, Ti.a, J. Immunol. Methods, 65:55, 1983.

2. Jacobs, J.?., J. Sad. Cancer Insc. , 3^:231, 1955.

BAD ORIGINAL

9/Q38-1-C1

The following examples further illustrate the present invention ar.d will enable others skilled in the art to understand it more completely. It should be understood, however, thac the invention is not limited to the particular examples given below.

APO00189

BAD ORIGINAL ixamc.e ό - 11-Oir.vdro-ll· hvl -oH-dlovrldo ί 3.2-5:2¹ .2' . - e ’ '1.4 ’ diazeo ir. ό - thione

A mixture of 2.66g (0.01 mol) of 5,ll-dihydro-ll-echyL-5-!sethyL-0Hdipvridoί3,2-b:2'3¹ -e]ί1,4idiazepin-6-one and 2.10g (0.005 mol) of Lawesson's reagent (2,4-5is(4-mechoxyphenyl)-1.3-dithia2, 4-diphosphetane-2 Λ-disulfide) in 50inl of toluene was refluxed for 2 1/2 h. The solvent was then removed in vacuo and water was added to the residue. The product was extracted with ethyL acetate, dried (anhydrous sodium sulfate; and concentrated in vacuo. Purification was effected on a silica gel column using methylene chloride as the first eluent, followed by cchvl acetate/hexane (1:4). Removal of che solvent in vacuo gave 2.20g (74¾ of theory) of 5,11-dihydro-11-ethyl-5-methyl-6Hdipyridof3,2-b:2 ' ,3'-e1[1,41diazapin-6-thione as a yellow powder which was recrystallized from 10¾ hexane/ethvl acetate to provide l.lg of yellow needles, m.p. 157-158’C.

The resulting conpound was tested using the Reverse Transcriptase Inhibition Assay (at 10 ug/ml) and T-Cell Culture Assay (at 3 -ug/ml) and demonstrated 100% inhibition in both tests.

BAD ORIGINAL

Precar at icn of startinc comround

5.11-Dihvdro-6H-dipvrIdo> 3.2-b:2' . 3 ' -άj Γ1.4'dlazeoin-6-one

a) 2-Chloro-N-< 2-chloro-3 -pyrldinvl) - 2 -pvrldlnecarboxamide

In a three-necked round-bottomed flask, fitted with an efficient reflux condenser, mechanical stirrer and dropping funnel, were placed 215 g < 1.672 mol) of 3-araino-2-chloropyridine , dissolved in a mixture of -*C0 ml dioxane, 500 ml cyclohexane and 130 ml pyridine. The solution of 299.2 % (1.7 moi) of freshly prepared 2-chloro-3-pyridinecarboxylic acid chloride in 200 mi dioxane was added at such a rate as to keep the vigorous reaction under control. Thereafter, the reaction mixture was allowed to cool to room temperature and the resulting crystalline precipitate was filtered off and washed successively with cyclohexane and ether.

The dark brown product was dissolved in 5 1 of a 3% aqueous solution of sodium hydroxide. The resulting solution was treated with charcoal, suction filtered, and the filtrate was acidified by addition of 50% aqueous acetic acid. The resulting precipitate was collected by filtration and thoroughly washed with water. After being dried overnight in a stream of nitrogen at room temperature the almost colorless product had a m.p. of 156-159‘C and was sufficiently pure for further reactions. The yield was 376.0 g (3i% of theory).

AP 0 0 0 1 8 9 /O38-I-C1

BAD ORIGINAL

b)

- Ohio ro - 3 - ;·-··,···1ϊγ.·.·’? - 2 - ' ' - T.e thoxuohenvl'msthvl ’ amino ί

- 2 - ovridinecavooxamide

13.1 g <0-03 mol) of the product obtained in s:a? a) were dissolved in 20 tai of xylene, and che resulting soLucion was admixed with 13.8 g (0.1 mol) of p-methoxybenzylamine. Thereafter, the mixture was refluxed for two hours. The reaction mixture was then evaporated in vacuo. and the residue was purified by column chromatography on silica gel (0.2-0.5 mm) using dichloromethane/ethyl acetate 10/1 (v/v) as an eluent. Colorless crystals, melting at 122-I2i°C (after rscrystailiracion from acetonitrile). The yield was 17.2 g (93% of theory).

c) 5. 11-Dihvdro-ll-> (6-mechoxvphenyl)methyll -6H-di.pvrl,do(3,2-5:2¹ .3’-e Η 1.1'diateoin-8-one

16.7 g (Q.Q153 mol) of the product obtained in step b) were dissolved in 150 ml of absolute dioxane, and the resulting solucion was admixed with

6.7 g (0.¼ mol) of a 50% dispersion of sodium hydride in mineral oil.

Thereafter, the mixture - while protected against the external atmosphere by a low flow of nitrogen - was refluxed until no starting material could be detected by TLC. The surplus of sodium hydride was decomposed by cautious addition of 10 ml of a mixture of methanol and tetrahydrofuran (50/50 v/v). The reaction mixture was neutralized by addition of acetic acid and then was evaporated in vacuo. The residue was purified by column chromatography on silica gel (0.2-0.5 mm) using successively dichloromethane/ethyl acetate 10/1 (v/v) and dichloromethane/ethyl acetate 1/1 (v/v) as eluents. The crystalline product obtained by evaoortion'of suitable fractions was recrystallited from acetonitrile and

9/038-1-CL

-· - —S

BAD ORIGINAL z r ccar.c 1. The pzccuct had a a.?, of

5.11- i.h'’c-.j - IT / -»-methoxypr.eny T mechy ; 1. T dazes in-m-or.e . The yield was 10 (Mackerev-Nagel, Polygram^3, SIL G/UV,_5u,

213-215’C and vas identified as

Τ'· oH-dipyvido (3. 2 - b : 2 ’ , 5' ·*.

g (63% of theory). 3.-0.7 precoated plastic sheets for TLC:

dichlo rome zhar.e/ethyL acetate 1/1 v/v)

d) 5. 21 · llkvdro -GH-diovrldoi 3,2-6:2¹ .3’- el (ΙΛ] diazapin-6-one

10.0 g ,0.3 mol) of the product obtained in step c) were dissolved in 50 ml of trifluoroacetic acid whereby the .mixture became slightly warm. Thereafter, the reaction mixture was stirred at 60*C for 1 hour. No starting material could be detected by TLC ac that time. The mixture was then evaoorated in vacuo. The residue thus obtained was thoroughly stirrec with 3.3% aqueous ammonia and then was filtered by suction. The raw product was recrystallized from 150 el of dimethyl sulfoxide to provide colorless crystals of m.p. of > 340*C. The yield was 4.8 g (75% of theory).

The product was identified as 5.ll-dihydro-6H-dipyrido[3.2-b: 2' , 3'-eJ [L,T diazepin-6-one .

e) Following step c) lOg of the resulting compound can instead be reacted with trifluoroacetic acid, stirred for one hour at room temperature, the acia removed vacuo and the residue stirred for one hour with 0.3¾ ammonia The solic was filtered and dried to give 6.7g of the 5-methyl substituted derivative. Z.Og of a 50¾ dispersion of nah in mineral oil was added to

5.75g of the 5-Methyl derivative in 100 ml dimethylformamide. After cessation of hydrogen evolution the mixture was heated to 50 degrees C for 30 minutes and the mixture stirred overnight at room temperature. Excess sodium hydride was decomposed by the addition of ice followed by water. The product was extractec with eather, dried (anhydrous sodium sulfate) and evaporated to give 4.5g of 5,-11-dihydro-11-ethyl-5-methyl-6-h dipyrido (3,2-8:2¹,3'-E) (1,4)diazepin-6-one M.P. 130-132 degrees C.

AP 0 0 0 1 8 9

BAD ORIGINAL

The following compounds were prepared analogously to Example 1 (preparation of starting materials) and according to methods C and D (for Z - 0) and analogously to Example 1 (method Ξ) (for Z - S), and ware tested using the Reverse Transcription Inhibitory Assay described above.

I

Z is an oxygen atom unless the compound is noted to be a thiolactam.

The compounds are unsubstituted at the 2, 3_Z 4, 7, 8 and 9 positions unless noted in the column headed OTHER

	η			Inhib.®	M.P.*C
Example	R¹	R OTHER		10 ug/ml
2	H	-CH₂Ph-4-OMe		81¾	209-210
3	Me	-CH₂Ph		86¾	-
4	Me	-CH₂PH-4-OMe		45¾	120-121.5
5	Me	-ch₂ch₂f		96¾	117-118
6	H	-Ph		71¾	220-222
7	H	Et 4-Et		100¾	212-214
8	Me	Et 8-C1		94¾	105-106
9	Me	Et 4-Me		98¾	157-159
10	Me	-COCH-		73¾	138-143
11	-CH,SCH^	Et		93¾	118-120
12	H	Et 2,3-di	Me	100¾	212-214
13	H	Et 9-Me		86¾	244-247
14	H	Et 2-Me		91¾	263-266
15	H	Et 2,4-di	Me	100¾	210-211

31a

BAD ORIGINAL

Znhlb.l _M.p.»_c

OTHER 10 pg/ml

H	Et	3-Me	95¾
Me	Et	3-Me	96¾
H	-COCH.		60¾
Ma	Et	2,9-di Ma	98¾
H	Et	2,4,9-tri Me	100¾
Me	Et	2-Me	100¾
H	Et	3,4-di Ma	100¾
Me	Et	3,4-di Me	99¾
Me	Et	9-Me	96¾
-coch₃	Et		9 6¾
Me	-CH₂SCH₃		99¾
H	Et	3-Cl	92¾
Me	Et	3-C1	99¾
-CH^Ph			37¾
Me	-CH₂CH=CH₂		99¾
H	Et	2-C1	96¾
Me	Et		100¾
Me	-CH₂CsCH		94¾
Me	Et	2-C1	100¾
H	Et	7-Ma	80¾
H	Et	8,9-di Ma	42¾
H	Et	8-Me	89¾
H	Et	4-Cl	100¾
H	Et	4-OMe	96¾
H	Et	4-Et	100¾
H	Et	8-Br	-
Me	Et	3-NO₂	-
Me	Et	3-NH₂	-
H	i-Pr	4-Me	-
Me	Et	2,4-di Ma	-
Ma	Et	7-Ma	-
H	Et 4-Me	(thiolactam)	-
H	Et 4-OH	(2HBr salt)	-
Me	Et	2-OMe	—

not avail.

94-36 >215 (dec). 100-102 228-230 124-126 265-266 119-120

79-93

123- 124.5 109-110

217- 218

124- 125 169-170

93-95 252-254 143-145 169-170

125- 126 193-194 204-206 182-183 184-186 156-157

218- 219 233.5-235.5

154-156

235-240

188-189

151-153

122-124

158-159

295-296

116-118

AP 0 0 0 1 8 9

31b

EXAMPLE A

A-l			A-2
Ingredients ¹	Ouanc	itv	Ingredients Quantity
Active compound	50	mg	Active compound	50 mg
Starch	100	mg	Dicalcium Phosphate	150 mg
Microcrys, Cellulose	90	mg	Microcrys.' Cellulose	90 mg
Sodium Starch Gluctate	10	mg	Stearic acid	5 mg
Magnesium Stearate	2	mg	Sodium Starch Glycolate	10 mg
Fumed colloidal silica	1	mg	Fumed colloidal silica	1 mg
The compound of Example	2 is	: blended	inco a powder mixture with	che

Quantity

500mg

L-5§

0.1% by weight q.s. Co 100ml ith the water and thereafter the xing is continued until the solution premixed excipient materials as identified above vith the exception of the Lubricant. The lubricant is then blended in and the resulting bLend· compressed into cablets or filled inco hard gelatin capsules.

EXAMPLE 3

Parenteral Solutions

Ingredients Active compound

Tartaric acid

3enzyl Alcohol Vater for injection

The excipient materials are mixed w active compound is added. Hi is clear. The pH of this solution is adjusted co 3.0 and is then

AP 0 0 0 1 8 9

9/038-l-Cl bad ORIGINAL filtered into the atorooriate vials or amooules and cr

'.13 sterilized b :

Nasal Solutions

Ingredients

Active compound

Citric acid

Benzalkonium chloride

EDTA

Pol yv i ny1a 1c oho 1

Water

Quantity lOOmg

1.92s

0.025% by weight 0.1 % by weight 10% by weight

q.s. to 100ml

The excipient materials are mixed with the water and thereafter the active compound is added and mixing is continued until the solucion is clear. The pH of this solucion is adjusted co 4.0 and is then filtered into the appropriate vials or ampoules.

9/038-l-.Cl

BAD ORIGINAL

Claims

CLAIMS:

1 to 3 or 6.

1 * 1 . wherein B has the same definition as for B with the exeption of hydrogen and X is the radical of a re1’ active ester, a halogen atom, the group OSO^OB , the methanesulfonyloxy or ethanesulfonyloxy group or an aromatic sulfonyloxy group,

C'. for preparing compounds in accordance with Formula I in 2 8 which B through B are as defined with respect to Formula I and B^ is other than hydrogen, by reacting a compound of Formula IV as defined above with a compound of Formula V as defined above in the presence of an amine or of an alkali carbonate or bicarbonate.

D. for preparing a compound of formula I in which B? is other than alkanoyl, hydroxyalkyl or alkoxycarbonyl and 13 8

B and B through B are as defined with respect to Formula I, by converting a compound of Formula I in which B^ is hydrogen into the corresponding metal salt of general Formula IVa or, when B^ is hydrogen of Formula VIb

AP 0 0 0 1 8 9

2H⁺

Via VIb

---—

BAD ORIGINAL wherein R ia as hereinbefore defined and M represents an alkali metal, or M represents the group MgHal wherein Hal is a chlorine, bromine or iodine atom, and subse- quently alkylating with a compound of the general For- mula VIII

R² ' x VIII

1 2 with the proviso that when Z is oxygen and R* and R are the same or different and are hydrogen or straight chained or branched alkyl of 1 to 5 3 8 carbon atoms at least one of R through R is other than hydrogen, or a pharmaceutically acceptable acid addition salt thereof.

BAD ORIGINAL

1. A compound of the formula wherein.

Z is oxygen or sulfur;

R^x is hydrogen, alkyl or fluoroalkyl of 1 co 5 carbon acoms.

crihalomechyl, alkenyl or alkynyl of 3 co 5 carbon atoms,

2 · · 2 wherein S has the same definitions as for S with the exception of alkanoyl, hydroxyalkyl, or alkoxycarbonyl and X is as defined above

E. for preparing a compound of Formula I in which Z is 1 8 sulfur and R through R are as defined with respect to Formula I, by reacting a compound .of Formula I in which 2 is oxygen with a sulfurating agent, and wherein in each of the foregoing processes any reactive group may protected as desired or necessary, and thereafter as desired.

a) deprotecting any protected group,

b) hydrolysing a nitro group to an amino group,

c) acylating an amino group to form an alkanoylamino group

d) alkylating an amino or aminoalkylgroup to form a mono- or dialkylamino group or mono- or dialkylaminoalkyl group,

e) acylating the 11-position nitrogen atom of a compound of Formula I is which R² in hydrogen, preferably subsequent to process E, and therafter isolating the compound of Formula I as such or as a pharmaceutically acceptable acid addition salt thereof. BAD ORIGINAL

2-halo-propen-l-yl, or aikoxyalkyl or alkylthioalkyl of 2 co 3 carbon acorns;

r2 is alkyl or fluoroalkyl of 1 co 4 carbon acorns, alkenyl or alkynyl of 2 co 4 carbon acorns, alkoxyalkyl or alkylthioalkyl of 2 co 4 carbon acorns, alkanoyl of 2 co 3 carbon acorns, hydroxyalkyl of 2 co 4 carbon acoras, arylmethyl (wherein che aryl rooiecy is phenyl or chienyl, which is eicher unsubstituted or substituted by methyl, hydroxyl, methoxy or halogen), phenyl (which is eicher unsubscicuced or subscicuced wich mechyl, mechoxy, hydroxyl or halogen) or alkoxycarbonylmechyl wherein che alkoxy moiecy concains 1 co 5 carbon acorns;

R.3, sA, and R? are each independently hydrogen or mechyl, wich the proviso chat at lease one of chese substituents is hydrogen, or R^ i_sechyl, propyl or bucyl wich che remaining cwo substituents being hydrogen;

r a

R , R', and R are each independently hydrogen or methyl, with the proviso chat at least one of these substituents is hydrogen, or R^ is echyl, propyl or butyl with the remaining two substituents being hydrogen;

or a pharmaceutically acceptable acid addition salt thereof.

2. A compound of formula I. as sac torch in claim 1, wherein,

Z is oxygen or sulfur;

3Λ is hydrogen, alkyl or fluoroalkyl of 1 Co 5 carbon acorns, crihalomechyl, alkenyl or alkynyl of 2 co 4 carbon acorns,

2-halo-propen-l-yl, arylmechyl (wherein the aryl moiecy is phenyl or thienyl or furanyl, which is eicher unsubscicuced or substituted by methyl, mechoxy or halogen), alkanoyl of 2 to 3 carbon acoms, or alkoxyalkyl or alkylchioalkyl of 2 co 4 carbon acoms;

is alkyl or fluoroalkyl of 1 to 5 carbon atoms, alkenyl or alkynyl of 2 to 5 carbon atoms, alkoxyalkyl or alkylchioalkyl of 2 co 4 carbon acoms, alkanoyl of 2 co 4 carbon acoms, hydroxyalkyl of 2 co 5 carbon atoms, arylmechyl (wherein che aryl moiecy is phenyl, Chienyl or furanyl, which ia eicher unsubscicuced or subscicuced by alkyl or alkoxy of 1 to 3 carbon acoms, hydroxyl or halogen), phenyl (which is eicher unsubscicuced or subscicuced by alkyl or alkoxy of 1 co 3 carbon acoms hydroxy or

AP 0 0 0 1 8 9

3 through 3 are as defined with repect to formula I and 3^ is other than hydrogen, by converting a 5,ll-dihydro-6H-dipyrido (3,2-b:2',3'-<][1,4] diazepin-6-one of the Formula IV (IV) wherein 5 through B are as defined above, into the corresponding 5-alkali or alkline earth metal compound and subsequently reacting the alkali metal compound with a reactive ester of the Formula V

B¹'* (V)

3 9

R through R are each hydrogen;

or a pharmaceutically acceptable acid addition salt thereof.

3. A compound of formula I, is set forth in claim 1, wherein, 2 is oxygen or sulfur;

R^x is hydrogen, alkyl or fluoroalkyl of 1 to 4 carbon atoms or allyl;

R is straight or branched alkyl or fluoroalkyl of 1 to 4 carbon atoms, allyl or benzyl and,

4. A method of preparing a compound as defined in any one of claims 1 to 3, which comprise carrying out one of tbe following methods;

BAD ORIGINAL

λ. for preparing compounds in accordance with formula 1 ia which is ocher than hydrogen, cyciising. a carboxylic acid amide of the general formula II fi²'

13 3 wherein 3 and 3 through 3 have the same meanings as set 2 · forth with respect to formula I and 3 has the same definitions as with the exception of hydrogen.

AP 0 0 0 1 8 9

BAD ORIGINAL for producing compounds in accordance with Formula lb in which 3 and 3 through 3 have the same meanings as set forth with respect to Formula I, hydrolytically cleaving the aryl methyl group in a compound of general Formula III

13 8 in which 3 and 3 through 3 are as. defined above and Ar is a phenyl or 4-methoxyphenyl group,

C. for preparing compounds in accordance with Formula I in which 2 8

5. A method as set forth in claim 4, in para A thereof.

6 8 I 0 0 0 dV

6. A compound of formula I as set forth in any one of claims 1 to 3, whenever prepared by a method as set forth in claim 4 or 5.

6 8 V 0 0 0 dV

BAD ORIGINAL

7. A pharmaceutical composition suitable for preventing or treating HIV-1 infection which comprises a prophylactically or therapeutically effective amount of a compound of formula I, as set forth in claims 1 to 3 or 6, and a pharmaceutically acceptable carrier.

8. A method of preparing a pharmaceutical composition which comprises mixing a compound as defined in any one of claim 1 to 3, or 6 or prepared by a method according to claim 4 or 5, with a pharmaceutically acceptable carrier or excipient.

9. Use of a compound as defined in any one of claims 1 to 3 or 6, or prepared by a method according to claim 4 or 5 in a method of preparing a pharmaceutical composition

9/038-1-C1

9/038-1-Cl

-_- bad original

CO halogen) oc aIkouycarbouyImethyl werehi che alkoxy moiety contains : carbon atoms; and,

F.·⁵ through R® are each hydrogen; or, one of R^J through R is alkyl of 1 to i carbon atoms, alkoxy or alkylthio ο; 1 to u carbon atoms, aikoxycarbonyl of 2 to t carbon atoms, hvdroxyalkyl of 1 to * carbon atoms, alkanoyl of 2 to 4 carbon atoms, alkanovloxy of 2 to t carbon atoms, alkanoylamino of 1 to ύ carbon atoms, aminoalkyl of 1 to i carbon atoms, alkoxycarbonylalkyl -«-herein the aikoxy and alkyl moieties each contain 1 to 2 carbon atoms, carboxyalkyl of 2 to j. carbon atoms, mono- or di-alkylamino wheein each alkyl moiety contains 1 to 2 carbon atoms, cyano, nitro, hydroxyl, carboxyl, amino, mono- or di-alkylaminoalkyl wherein each alkyL moiety concains 1 co 2 carbon atoms, atido or halogen, with the ocher five substituents being hydrogen;

or

R^, r\ and R® are each independently hydrogen or alkyl of 1 to 3 carbon atoms, with the proviso that ac least one of these substituents is hydrogen, or one of R?. r⁴ and r5 £_s bucyl with the remainingtwo substituents being hydrogen; and,

R®, R' , and R^ are each independently hydrogen or alkyl of 1 to 3 carbon atoms, with Che proviso that at least one of these substituents is hydrogen, or one of R° , R' and R° is butyl with the remaining two substituents being hydrogen;

10. A pharmaceutical composition for preventing or treating HIV-1 infection comprising a compound as set forth in any one of claims 1 to 3 or 6.

bad ORIGINAL

11. Use of a compound of formula I in a method for preventing or treating HIV-1 infection which comprises administering, to a human being exposed to or infected by HIV-1, a prophylactically or therapeutically effective amount of a compound of formula I, as set forth in any one of claims

12. A composition as claimed in claim 7 or 10 contained in a package which carries or contains instructions for the use thereof in preventing or treating HIV-1 infection.