NZ261740A

NZ261740A - Pharmaceutical compositions for treating hiv

Info

Publication number: NZ261740A
Application number: NZ261740A
Authority: NZ
Inventors: Albert Stanley Tyms; Debra Lynn Taylor
Original assignee: Merrell Dow Pharma
Priority date: 1993-02-22
Filing date: 1994-01-18
Publication date: 1996-10-28
Also published as: HU9502451D0; WO1994019008A1; CN1118142A; IL108696A; JPH08509469A; CA2155129C; GB9303518D0; KR100293299B1; HUT72493A; KR960700743A; NO953272D0; AU6091894A; ZA941036B; NO307738B1; IL108696A0; NO953272L; CA2155129A1; EP0684836A1; MXPA94001311A; AU679497B2

Description

<div class="application article clearfix" id="description"> New Zealand Paient Spedficaiion for Paient Number £61 740 New Zealand No. 261740 International No. PCT/US94/00710 Priority Date(s):.. ...aaL Comptete Specification F.!vd: Class: (3) Hfc>.l.K38.|€Sf Publication Date:?'!f..P.^!.i??® P.O. Journal No: NEW ZEALAND PATENTS ACT 1953 COMPLETE SPECIFICATION Title of Invention: Combinations of retroviral inhibitors Name, address and nationality of applicant(s) as in international application form: MERRELL DOW PHARMACEUTICALS INC, ;. of 2110 East Galbraith Road, Cincinnati, Ohio 45215-6300, United States of America/ „ us wo 94/19008 -1- 2&JLZ.4 0 5 combinations of retroviral inhibitors 10 This invention relates to novel combinations of castanospermine esters and short peptide an£s?.ogs that act effectively in their combination in the treatment of 15 retroviral infections, particularly in the treatment of acquired immune deficiency syndrome(AIDS) and related human immunodeficiency viral (HIV) infections. BACKGROUND OF THE INVENTION Intense research over the last several years has been devoted to develop treatments and cures for retroviral viral 25 infections in humans and in animals, and particularly for acquired immune deficiency syndrome (AIDS) and AIDS related complex (ARC). Today the public realizes the severe health risk posed by the AIDS retrovirus and that the incidence of ARC and AIDS in humans is increasing at an alarming rate. 30 Furthermore, survival beyond 5 years for those who have contracted AIDS remains remote. Further, AIDS patients whose immune systems have been seriously impaired by the infection, suffer from numerous opportunistic infections and proliferative disease including Pneumocystis carninii 35 pneumonia and Kaposi's sarcoma. No cure for AIDS is known and current treatments are largely without adequate proof of efficacy and have numerous untoward side effects. Due to the severity of the disease, and with death the resultant WO 94/19008 PCT/US94/00710 -2- outcome from the disease, fear of AIDS has resulted in social ostracism and discrimination against those having or suspected of having the disease. ^ The AIDS virus belongs to a general class of viruses known as retroviruses. As a class, many of the known retroviruses are also oncogenic or tumor causing. Indeed the first two human retroviruses discovered, denoted human T-cell leukemia virus type I and type II or HTLV-I and II, were found to cause rare leukemias in humans after infection of T-lymphocytes. The third such human virus to be discovered, HTLV-III, now referred to as HIV, was found to cause cell death after infection of T-lymphocytes and has been identified as the causative agent of acquired immune 15 deficiency syndrome (AIDS) and AIDS related complex (ARC). Retroviruses are a class of ribonucleic acid (RNA) containing viruses that replicate by using a reverse transcriptase activity to form a strand of complementary DNA 20 (cDNA) from which a double stranded, proviral DNA is produced. This proviral DNA is then incorporated into the chromosomal DNA of the host cell making possible viral replication by transcription of this integrated DNA and translation of viral messenger RNA into proteins. 25 Replication of the virus occurs by synthesis of viral genomic RNA and its assembly with glycosytated and non-glycosylated viral proteins to form new viral particles. Maturation of virions at the cell surface results in the release of infectious virus progeny. 30 Retroviral proteins are generally synthesized as polyproteins and virus encoded proteases are required to cleave the precursor polyproteins to form the viral enzymes and structural proteins. For example, the gag and gag-pol 35 precursor polyproteins of retroviruses are synthesized as precursors of viral encoded enzymes and non-glycosylated structural proteins. Similarly, the envelope protein of HIV WO 94/19008 PCT/US94/00710 "3~ is a 160 kDa highly glycosylated precursor glycoprotein. The envelope proteins are cleaved by a host-cell protease to give a 120 kDa external glycoprotein (gp 120) and a transmembrane glycoprotein (gp 41). The gp 120 protein ^ contains a high affinity binding site that recognizes the CD4 ligand on CD4-positive human T-helper cells, the known receptor for this virus. The retroviral proteases also show certain commonality 10 by their inhibition by aspartyl protease-specific inhibitors, Iyoko, et al. Nature 329, 654-67. Similarly, amino acid sequencing of the retroviral proteases show they possess sequence homology. Because of their mutual structural and functional characteristics and their obligate function, the aspartyl proteases serve as a potentially interesting therapeutic target for intervention. The correctly processed envelope glycoproteins of the retroviruses play an important role in the virus life cycle, 20 which also offers a possible target for clinical intervention. The envelope glycoproteins serve a role in both the initial interaction of the virion and the target host-cell and in the subsequent fusion of the viral envelope and host-cell membranes during penetration. Certain esters 25 of castanospermine are useful in interfering with the processing of the viral envelope glycoproteins and thereby in preventing the initial virus-host cell interaction and subsequent fusion. 35 WO 94/19008 -4- PCT/US94/00710 The applicants have discovered that the combination of aspartyl protease-specific inhibitors (formula. II) with glycoprotein processing inhibitors such as castanospermine ^ derivatives (formula I) result in significant improvements in the inhibition of the HIV virus that would not be expected. SUMMARY OF THE INVENTION ^ The pharmaceutical compositions consist of the glucosidase inhibitor of formula I and a viral aspartyl protease inhibitor of formula II. The pharmaceutical compositions of formula 1 and formula 15 II, and their pharmaceutical^ acceptable addition salts, are novel and possess valuable pharmacological properties. Often these compositions can act synergistically to effectively inhibit HIV activities, and therefore, can be used in the prophylaxis or treatment of viral infections, 20 particularly infections caused by HIV. Objects of the present invention are the compositions of formula I and II, and their aforementioned salts, for use as therapeutically active substances, medicaments containing 25 said compounds and salts, their manufacture, and the use of said compositions and salts in the control or prevention of illnesses, especially in the treatment or prophylaxis of HIV infections. 35 WO 94/19008 PCT/US94/00710 Compounds of formula I comprise structures of the following formula: 15 wherein R, Ri and R2 are independently hydrogen, Ci-14 alkanoyl, C1-C14 alkenoyl, cyclohexanecarbonyl C1-6 alkoxyacetyl, 20 25 30 35 naphthalenecarbonyl optionally substituted by methyl or halogen; phenyl(C2-6 alkanoyl) wherein the phenyl is optionally substituted by methyl or halogen; cinnamoyl; pyridinecarbonyl optionally substituted by methyl or halogen; dihydropyridine carbonyl optionally substituted by Ci_io alkyl; thiophenecarbonyl optionally substituted by methyl or halogen; or furancarbonyl optionally substituted by methyl or halogen; Y is hydrogen, C1-4 alkyl, C1-4 alkoxy, halogen, trifluoromethyl, Ci_4 alkylsulfonyl, C1-4 alkylmercapto, cyano or dimethylamino; Y' is hydrogen, C1-4 alkyl, C1-4 alkoxy, halogen or it is combined with Y to give 3,4-methylenedioxy; Y" is hydrogen, Ci_4 alkyl, C1-4 alkoxy or halogen; with R, Ri and R2 being selected in such a way that at least one of them, but WO 94/19008 PCT/US94/00710 10 15 20 25 30 35 -6- not more than two of them, is hydrogen; or a pharmaceutical^ acceptable salt of these compounds. The Ci-i4 alkanoyl groups referred to above can be straight- or branched-chain or cyclic and can be exemplified by formyl, acetyl, propionyl, butyryl, isobutyryl, cyclopropanecarbonyl, hexanoyl, octanoyl and decanoyl. The C1-14 alkenoyl groups referred to above can be straight- or branched-chain or cyclic but have at least one carbon-carbon double bond as exemplified, propenoyl, butenoyl, isobutenoyl, hexenoyl, octenoyl and decenoyl. The Ci_6 alkoxyacetyl referred to above can be methoxy-acetyl, ethoxyacetyl and butoxyacetyl. The halogens referred to above can be exemplified by fluorine, chlorine, bromine or iodine. The C2-6 alkanoyl groups referred to above can be exemplified by acetyl, propionyl, butyryl, isobutyryl, and hexanoyl. The Ci_4 alkyl groups referred to above, whether alone or as part of an alkoxy, an alkylsulfonyl or an alkylmercapto group, can be straight- or branched-chain alkyl groups containing up to 4 carbon atoms. Examples of various such groups are methyl, ethyl, propyl, butyl, methoxy, ethoxy, butoxy, methylsulfonyl, ethylsulfonyl, methylmercapto and ethylmercapto. The phenyl(C2-6 alkanoyl) groups referred to above can be exemplified by benzeneacetyl and benzenepropionyl. The various naphthalenecarbonyl, pyridinecarbonyl, thiophenecarbonyl and furancarbonyl groups referred to above include the various position isomers and these can be exemplified by naphthalene-1-carbonyl, naphthalene-2-carbonyl, nicotinoyl, isonicofcinoyl, N-methyl-dihydro-pyridine-3-carbonyl, thiophene-2-carbonyl, thiophene-3-carbonyl, furan-2-carbonyl and furan-3-carbonyl. The naphthalene, pyridine, thiophene and furan groups can be optionally further substituted as indicated above. WO 94/19008 PCT/US94/00710 -7- Preferred compounds of the present invention are those wherein R, Ri and R2 are 1 or 2 alkanoyl, alkenoyl, or benzoyl groups with the benzoyl substituted by Y, Y' and Y" as described above, especially a C1-4 alkanoyl or a benzoyl optionally substituted with an alkyl or halogen. More preferred are those compounds of formula 1 wherein r>r.c of R, Ri and R2 is alkanoyl or benzoyl, especially a C1-8 alkanoyl, Ci_8 alkenoyl, or a benzoyl optionally substituted with an alkyl or halogen, and the others 3re hydrogens. Even more preferred are those compounds of formula 1 wherein one of R, Ri and R2 is a Ci_8 alkanoyl, C1-0 alkenoyl, or a benzoyl optionally substituted with an alkyl or halogen, especially a methyl, bromo, chloro, or fluoro group, and the others are hydrogens. 10 15 20 25 30 35 Most preferred grouping are those compounds of formula 1 wherein Ri is a Ci_8 alkanoyl, C1-8 alkenoyl, or benzoyl optionally substituted with an alkyl or halogen, especially a methyl, bromo, chloro, or fluoro group, most especially a methyl, bromcr, chloro, or fluoro group at the para position, and wherein R and R2 are each a hydrogen. A most especially preferred grouping can be designated by the substituents represented by the following compounds: 1. [lS-(la,6B,7a,8B,8aB)]-octahydro-1,6,7,8-indolizinetetrol 6-benzoate: 2. (IS-(la,6B,7a,8 8,8aB)]-octahydro-1,6,7,8-indolizinetetrol 7-benzoate: 3. [lS-(la,6B,7a,88,8aB)]-octahydro-1,6,7,8-indolizinetetrol 6-(4-methylbenzoate): 4. [IS-(la,63,7a,8B,8a8)]-octahydro-1,6,7,8-indolizinetetrol 7-(4-bromobenzoate): WO 94/19008 PCT/US94/00710 -8- 5. [lS-(la,6B,7a,88,8a8)]-octahydro-l,6,7,8-indolizinetetrol 6,8-dibutanoate: 6. A castanospermine ester of Claim I which is [lS-(la,6B,7a,88,8a8)]-octahydro-1,6,7,8-indolizinetetrol 6-butanoate: 7. [ lS-(la,6B,7a,88,8aB)]-octahydro-l,6,7,8-indolizinetetrol 6-(2-£urancarboxylate): 8. [ lS-(la,6B,7a, 8B,8a8) ]-octahydro-l, 6,7,8-indolizinetetrol 7-(2,4-dichlorobenzoate) 9. [is-(la,6B,7a,88,8a8) ] -octahydro-1,6,7,8-indolizinetetrol 6-(3-hexenoate). I.0 . [is-(la,6s,7a,86,8as) ] -octahydro-1, 6 ,7 ,8-indolizinetetrol 6-octanoate. II. [lS-(la,6S,7o,86,8a6) ]-octahydro-1,6,7,8-indolizinetetrol 6-pentanoate. 10 15 20 25 Certain compounds are preferred. Amongst the preferred compounds of formula I is [1S-(la,6s,7a,88,8aB) ]-octahydro-l,6,7,8-indolizinetetrol 6-butanoate. Preparation, and preferred grouping of compounds of compounds of formula I are also taught in the US Patent No. 5,017,563, Issued May 21, 1991, which is herein incorporated by reference. Processes for the preparation of Castanospermine are also taught in US Patent No. 5,066,807, Issued November 19, 1991, which is herein incorporated by reference. 30 35 WO 94/19008 PCT/US94/00710 -9- Compound of the formula II comprise structures of the following formula: " I- CNHCH II O CNHCHC-CF^C—NReRK II II 1l 56 0 0 0 10 and the hydrates, isosteres and the pharmaceutically acceptable salts thereof wherein 15 20 25 30 a b c d e is zero or one, is Q, or B, B being CH? r^{-^(CH2)a-(0)b-(CH2)f.-Rj d with the proviso that B is other than 2~hydroxy-benzyl or j^alkoxybenzyl, is zero, or 1, 2 or 3, is zerc ot 1, is zero or 1, 2, 3, 4 or 5, is 1 or 2, is zero, 1 or 2, is (ch2)d, P2 is C1_6 alkyl, cyclopentyl, cyclohexyl, hydroxy F-^-P alkylene,, ri n-t, CH2CONHR4, or CH2CONHR4; with T being H or C(0)R4, 35 R is hydrogen, -CH2CHO, hydroxy Cj.g alkylene, C1_6 alkoxy C^galkylene, C^.g alkyl, phenyl, _J^jf}~(R3)d or Q, Rx is benzyloxy, C1-6 alkoxy, C1-6 alkyl, phenyl, benzyl, phenethyl, fluorenylmethylenoxy, 2-isoquinolinyl, PDL, WO 94/19008 PCT/US94/00710 -10- CH2N-(CH2)3pH2, (j)-(CH2)2-N-CH2(j:H2, NHS02R4, N(R4)(benzyl) , and N(R4)(PDL), with PDL being -(CH2)a-2->3-, or 4-pyridyl, or £-W-substituted benzyloxy with W being nitro, OH amino, Cj_6 alkoxy, or hydroxy CL_6 alkylene, or halogeno, R3 is C1-6 allenyl C1-6 alkoxy, C^.g alkoxy C^.g alkylene, hydroxy Cj^g alkylene, C1-6 alkyl, H, or OH, hydroxy C1_6 alkylene, C1_6 alkyl, oi (CH2}—CgH4—fV)e, and Z being CHO, C02R4, C02NHR4 oi {CH2-h-OR4, R6 is as defined for R5 with the proviso that Rg is other than H when Rg is H, and when R5 and Rg are taken together with nitrogen atom to which they are attached form a heterocyclic moiety of the formulae: 10 R4 is H, Cj^g alkyl, phenyl or benzyl, 30 WO 94/19008 PCT/US94/00710 -N(CH2)3CH2, (a) -11- -N(CH2)4CH2, -N(CH2)2OCH2CH2, (b) (C) 10 (d) ?3 I R,—Site) H N (f) or 15 -N(CH2)2N-CH2CH2, CH(O) (g) II if -NCH2C(CH2)2CH2, or -N-(CH2)2CCH2CH2, I 1 I I (h) (i) 20 25 30 35 R7 is CH2OR4 or C(0)NHR4, R8 is (H,OH) or =0. As is true for most classes of compounds found to be useful in the pharmaceutical industry, certain subgeneric groups and certain specific compounds are more preferred. Within the concepts of this invention, it is to be found that the preferred compounds are those wherein when R5 is H, then; Rt is benzyl, benzyloxy, 4-alkoxybenzyloxy, morpholyl, S02HN, {3-pyridyl)ethyl, isoquinolyl; P2 is methylamid, isopropyl, cyclopentyl, 2-(4,4-difluoro)-pyrrolidyl, 2-hydroxy-2-propyl, t-butyl; Px is piperonyl, 4-(benzyloxy)benzyl, 3-(benzyloxy)benzyl, (4-benzyloxy-3-methoxy)benzyl; WO 94/19008 PCT/US94/00710 -12- R6 is benzyl, piperonyl, CH2-pyridyl, 4-(benzyloxy)benzyl, morpholino, tetrahydroisoquinolyl, 4-(3-hydroxypropyl)benzyl, OH 20 2-(3-hydroxypropyl)benzyl, and or -CH(Y)(Z) 10 with Y and Z both being as generally defined, but particularly when Y is isopropyl, preferably in the D configuration, or phenyl and when Z is benzyloxymethylene, CHO, COOH, alkoxy or COOR4. When R5 is other than H it is preferred that R5 be methyl, 4-hydroxybutyl or 3-hydroxypropyl and that R6 be benzoxy or benzyl, and when R5 and R6 form a heterocyclic 15 moiety with the nitrogen attached thereto, the heterocycle is a perhydroisoquinoline of (f), -n Concho 0f(g), and morpholino of (c) be the heterocyclic moieties. The preferred specific compounds of formula II are those shown in the chart below and those products exemplified herein. 25 30 35 WO 94/19008 -13- PCT/US94/00710 10 15 20 25 30 35 A most especially preferred grouping can be designated by the substituents represented by the following compounds: 1. N-tertiary butyl-decahydro-2[R(R)-hydroxy-4~phenyl-3(S)—[[N(2-quinolylcarbonyl)-L-asparaginyl]amino]-butyl]-(4aS,8aS)-isoquinoline-3(S)-carboxamide; 2. [ 1 ( S) - [ [ 3 ,3-difluoro-2,4,-dioxo-l-[[4-(phenylmethyl)amino]butyl]aminocarbonyl]-2-methylpropyl]carboamic acid, phenylmethyl ester; or 3. N-[4-(N-benzyloxycarbonyl-L-valyl)amino-2,2- difluoro-1,3,-dioxo-5-(e-benzyloxy)phenyl-pentyl]-(0-benzyl)-D-valinol. The specific making and compounds of compounds of formula II are further taught in the International Application Published Under the Patent Cooperation Treaty, International Publication Number W0 92/12123, herein incorporated by reference. In the present invention, each compound in the compositions compounds of formula I and II may occur with asymmetric centers or may occur as racemates, racemic mixtures and as individual diastereomers, with all isomeric forms of the compounds being included in the present invention. The preferred compounds of formula I and formula II may be selected in any combination from one group select from formula I and one group selected from formula II. It is recognized that such combinations would include, for instance, a pharmaceutical composition comprising a compound of formula I: WO 94/19008 PCT/US94/00710 -14- ho 10 r2o Formula 15 ori wherein Ri is a Ci-8 alkanoyl, Ci-io alkenoyl, Ci-8 alkoxyacetyl, or benzoyl optionally substituted with an alkyl or halogen group; or a pharmaceutically acceptable salt thereof and a compound of formula II 20 CNHCH II 0 cnhchc-cf7c-nr,.r(. II II ^1 56 o o o Formula II 25 30 and the hydrates, isosteres and the pharmaceutically acceptable salts thereof wherein Rx is benzyl . a, oxy, S02HN, (3-pyridyl)ethyl, isoquinolyl, 4-alkoxy- benzyloxy, or raorpholyl, P2 is isopropyl, cyclopentyl, 2-(4,4-difluoro)-pyrrolidyl, 2-hydroxy-2-propyl, t-butyl, is piperonyl, 4-(benzyloxy)benzyl, 3-(benzyloxy)benzyl, (4-benzyloxy-3-methoxy)benzyl, when R5 is H, R6 is benzyl, piperonyl, CH2-pyridyl, 4-(benzyloxy)benzyl, morpholino, tetrahydroisoquinolyl, 4-(3-hydroxypropyl)benzyl, OH 35 2-( 3-hydroxypropyl )benzyl, and ^' or ~CH(Y)(Z) with Y and Z both being as'generally defined, but particularly when Y is isopropyl, preferably in the D configuration, or phenyl and when Z is benzyloxymethylene, WO 94/19008 PCT/US94/00710 -15- CHO, COOH, alkoxy or COOR4, when R5 is other than H it is preferred that R5 be methyl, 4-hydroxybutyl or 3-hydroxypropyl and that Rg be benzoxy or benzyl, and when R5 and R6 form a heterocyclic moiety with the 5 /—\ nitrogen attached thereto, it is preferred tha -mn ^ncho and morpholino be the heterocyclic moieties. It is further understood further subgroupings from the above combination may be made from the groups of formula I and II defined herein. 15 20 25 30 35 *1 r2 P1 R^-H, R6 benzyloxy isopropyl piperonyl benzyl benzyloxy isopropyl piperonyl piperonyl tor isopropyl 4-(benzyloxy)benzyl benzyl benzyloxy isopropyl 3-(benzyloxy)benzyl benzyl benzyloxy t-butyl 3-(benzyloxy)benzyl benzyl benzyloxy isopropyl (4-benzyloxy-3-methoxy)benzyl benzyl bensyloxy isopropyl (4-benzyloxy-3-methoxy)benzyl piperonyl benzyloxy 2-hydroxy-2-propyl 4-(benzyloxy)benzyl benzyl (3-pyridyl)ethyl isopropyl 4-(benzyloxy)benzyl -CH2-(3-pyridyl) isoquinolyl isopropyl 4-(benzyloxy)benzyl -CH2~(3-pyridyl) (3-pyridyl)ethyl isopropyl 4-{benzyloxy)benzyl -C2H5-(2-pyridyl) (3-pyridyl)ethyl isopropyl 4-(benzyloxy)benzyl -CH2-(2-pyr idyl) benzyloxy t-butyl 4-(benzyloxy)benzyl PCoJ^ MD) R, benzyloxy isopropyl (3-pyridyl)ethyl isopropyl isoquinolyl isopropyl benzyloxy isopropyl r5=h, r6 4-(benzyloxy)benzyl 4-(benzyloxy)benzyl -cho 4-(benzyloxy)benzyl cho 4-(benzyloxy)benzyl |JL-och3 'lor *1 p2 Pi r5-h, r6 benzyloxy isopropyl 4- (benzyloxy)benzyl (o) benzyloxy benzyloxy (3-pyridyl)ethyl isopropyl t-butyl isopropyl 1 1 1 (benzyloxy)benzyl (benzyloxy)benzyl (benzyloxy)benzyl 4-4- 4- H^C02H (benzyloxy)benzyl (benzyloxy)benzyl (benzyloxy)benzyl c§> %"C02-t-Ba benzyloxy isopropyl 4- (benzyloxy)benzyl benzyloxy isopropyl 4- (benzyloxy)benzyl T ilU C02H 4-0-methyl-benzyloxy isopropyl 4- (benzyloxy)benzyl benzyl benzyloxy isopropyl 4- (benzyloxy)benzyl -N^ ^0 r1 P2 pl r5-h, r6 benzyloxy isopropyl 4- (benzyloxy)benzyl 4-(3-hydroxypropyl )benzyl benzyloxy isopropyl 4- •(benzyloxy)benzyl 2-(3-hydroxypropyl )ben2yl OH benzyloxy isopropyl 4- (benzyloxy)benzyl r1 Pl r5» r6 benzyloxy isopropyl 4- (benzyloxy)benzyl Rs=CH3, R6= —o (3-pyc idyl)ethyl isopropyl 4- (benzyloxy)benzyl R5=CH3, R6=benzyl (3-pyridyl)ethyl cyclopentyl 4- (benzyloxy)benzyl R5=CH3, Rg=benzyl benzyloxy 2-(4,4-difluoro) pyrrolidyl 4- (benzyloxy)benzyl R5=CH3, R6=benzyl morpholyl isopropyl 4- (benzyloxy)benzyl R5=CH3, Rfi=benzyl *1 p2 pl R5» R6 (3 -pyridyljethyl isopropyl 4- (benzyloxy)benzyl R5= 4-hydroxybutyl Rg= benzyl benzyloxy isopropyl 4- (benzyloxy)benzyl R5= 3-hydroxypropyl R6= benzyl (3- -pyridyl)ethyl isopropyl 4- (benzyloxy)benzyl R5,Rfi= morpholyl (3-pyridyl)ethyl cyclopentyl 4- (benzyloxy)benzyl R5,Rg= morpholyl benzyloxy isopropyl 4- (benzyloxy)benzyl R5,Rg= -N^ ^NCHO (3- -pyr idyl)ethyl isopropyl 4- (benzyloxy)benzyl tetrahydro-isoquinolyl benzyl methylamide benzyl perhydroiso-quinoline-CONHt-Bu WO 94/19008 PCT/US94/00710 10 15 20 25 30 35 -21- DETAILED DESCRIPTION OF THE INVENTION The pharmaceutical compositions of the present invention are useful as inhibitors of retroviral proteases and cellular Y~glucosidase I which are required for virus replication, particularly HIV-1 and HIV-2, the prevention or treatment of infection by the human immunodeficiency virus (HIV), and the treatment of consequent pathological conditions such as the acquired immunodeficiency syndrome (AIDS) in mammals capable of being infected with HIV virus, Treating AIDS (preventing infection by HIV 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 pharmaceutical compositions of this invention are useful in preventing infection by HIV after suspected past exposure to HIV by, e.g., blood transfusion, accidental needle stick, or exposure to patient blood during surgery. For these purposes, the compounds of the present invention may be administered orally, parenterally (including subcutaneous injections, intravenous, intramuscular, transdermal, intrasternal injection or infusion techniques), by inhalation spray, or rectally, in dosage unit formulations containing convention non-toxic pharmaceutically acceptable carriers, adjuvants and vehicles. Thus, in accordance with the present invention there i further provided a method of treating and a pharmaceutical composition for treating HIV infection and AIDS. The treat ment involves administering to a patient in need of such treatment a pharmaceutical composition comprising a pharma ceutical carrier and a therapeutically effective amount of WO 94/19008 PCT/US94/00710 -22- a compounds of formula I and II of the present invention, or a pharmaceutically acceptable salt thereof. These pharmaceutical compositions may be in the form of 5 orally-administerable suspensions or tablets? nasal sprays; sterile injectable preparations, for example, as sterile injectable aqueous or oleagenous suspensions or suppositories) or they may be administered transdermally. When administered orally as a suspension, these compositions are prepared according to techniques well known in the art of pharmaceutical formulation and may contain microcrystalline cellulose for imparting bulk, alginic acid or sodium alginate as a suspending agent, methylcellulose as a viscosity enhancer, and sweetener/flavoring agents known in the art. As immediate release tablets, these compositions may contain microcrystalline cellulose, dicalcium phosphate, starch, magnesium stearate and lactose and/or other excipients, binders, extenders, disintegrants, diluents and lubricants known in the art. For oral administration the compositions of formula 1 can be formulated into solid or liquid preparations such as 25 capsules, pills, tablets, troches, lozenges, melts, powders, solutions, suspensions, or emulsions. The solid unit dosage forms can be a capsule which can be of the ordinary hard- or soft-shelled gelatin type containing, for example, surfactants, lubricants, and inert fillers such as lactose, 30 sucrose, calcium phosphate, and cornstarch. In another embodiment the compounds of this invention can be tableted with conventional tablet bases such as lactose, sucrose, and cornstarch in combination with binders such as acacia, cornstarch, or gelatin, disintegrating agents intended to assist the break-up and dissolution of the tablet following administration such as potato starch, alginic acid, corn starch, and guar gum, lubricants intended to improve the 10 15 20 WO 94/19008 PCT/US94/00710 -23- flow of tablet granulations and to prevent the adhesion of tablet material to the surfaces of the tablet dies and punches, for example, talc, stearic acid, or magnesium, calcium, or zinc stearate, dyes, coloring agents, and ^ flavoring agents intended to enhance the aesthetic qualities of the tablets and make them more acceptable to the patient. Suitable excipients for use in oral liquid dosage forms include diluents such as water and alcohols, for example, ethanol, benzyl alcohol, and the polyethylene alcohols, either with or without the addition of a pharmaceutically acceptably surfactant, suspending agent, or emulsifying agent. When administered by nasal aerosol or inhalation, these compositions are prepared according to techniques well known in the art of pharmaceutical formulation and may be prepared as solutions in saline, employing benzyl alcohol or other suitable preservatives, absorption promoters to enhance bioavailability, fluorocarbons, and/or other solublizing or dispersing agents known in the art. The compositions including formula 1 and 2 may also be administered parenterally, that is, subcutaneously, intravenously, intramuscularly, or interperitoneally, as injectable dosages of the compound in a physiologically acceptable diluent with a pharmaceutical carrier which can be a sterile liquid or mixture of liquids such as water, saline, aqueous dextrose and related sugar solutions, an alcohol such as ethanol, isopropanol, or hexadecyl alcohol, 30 glycols such as propylene glycol or polyethylene glycol, glycerol ketals such as 2,2-dimethyl-l,3-dioxolane-4-methanol, ethers such as poly(ethyleneglycol) 400, an oil, a fatty acid, a fatty acid ester or glyceride, or an acetylated fatty acid glyceride with or without the addition 35 of a pharmaceutically acceptable surfactant such as a soap or a detergent, suspending agent such as pectin, carbomers, methylcellulose, hydroxypropylmethylcellulose, or 15 20 WO 94/19008 PCT/US94/00710 -24- carboxymethylcellulose, or emulsifying agent and other pharmaceutical adjuvants. Illustrative of oils which can be used in the parenteral formulations of this invention are those of petroleum, animal, vegetable, or synthetic origin, 5 for example, peanut oil, soybean oil, sesame oil, cottonseed oil, corn oil, olive oil, petrolatum, and mineral oil. Suitable fatty acids include oleic acid, stearic acid, and isostearic acid. Suitable fatty acid esters are, for example, ethyl oleate and isopropyl myristate. Suitable 10 soaps include fatty alkali metal, ammonium, and triethanolamine salts and suitable detergents include cationic detergents, for example, dimethyl dialkyl ammonium halides, alkyl pyridinium halides, and alkylamines acetates; anionic detergents, for example, alkyl, aryl, and olefin sulfonates, alkyl, olefin, ether, and monoglyceride sulfates, and sulfosuccinates; nonionic detergents, for example, fatty amine oxides, fatty acid alkanolamides, and polyoxyethylenepolypropylene copolymers; and amphoteric detergents, for example, alkyl-beta-aminopropionates, and 2- 20 alkylimidazoline quarternary ammonium salts, as well as mixtures. When rectally administered in the form of suppositories, these compositions may be prepared by mixing 2 5 the drug with a suitable non-irritating excipient, such as cocoa butter, synthetic glyceride esters or polyethylene glycols, which are solid at ordinary temperatures, but liquidize and/or dissolve in the rectal cavity to release the drug. 30 The pharmaceutically acceptable addition salts, either cation or anion salts, are those salts that are not substantially toxic at the dosage administered to achieve the desired effect and do not independently possess 3 5 significant pharmacological activity. The conversion of the composition of formula I and II may independently or jointly be formulated as addition salts. Illustratively, cation WO 94/19008 PCT/US94/00710 10 15 20 25 30 35 -25- salts include those of alkali metals, as for example, sodium and potassium; alkaline earth metals, such as calcium and magnesium; light metals of Group IIIA including aluminum; and organic primary, secondary and tertiary amines, as for example, trialkylamines, including triethylamine, procaine, dibenzylamine, 1-ethenamine, N,N'-dibenzylethylenediamine, dihydroabiethylamine, N-(lower)alkylpiperidine, and any other suitable amine. Sodium salts are preferred. An acceptable acid addition salt may be carried out by treating such compounds in a conventional manner with an inorganic acid or example a hydrobromic acid, sulfphuric acid, nitric acid, phosphoric acid etc., or with an organic acid such as acetic acid, citric acid, maleic acid, fumaric acid, tartaric acid, methanesulfonic acid, p-toluenesulphonic acid. Dosage levels o£ the order of 0.02 to 5.0 or 10.0 grams per day of the composition are useful in the treatment or prevention of the above-indicated conditions, with oral doses two to five times higher. For example, infection by HIV is effectively treated by the administration of from 10 to 50 milligrams of the compound per kilogram of body weight from one to three times per day. It will be understood, however, that the specific dose level and frequency of dosage for any particular patient may be varied and will depend upon a variety of factors including the activity of the specific compounds employed, the metabolic stability and length of action of compounds in combination with each other, the age, body weight, general health, sex, diet, mode and time of administration, rate of excretion, drug combination the severity of the particular condition, and the host undergoing therapy. The present invention is also directed to combinations of the HIV protease-inhibitory compounds with one or more agents useful in the treatment of AIDS, such as, for example, with known antiviral agents suitable for treating HIV 1 and HIV 2 viral infections, e.g., a ester of O 94/19008 PCT/US94/00710 10 15 -26- castanospermine of formula I with a viral protease inhibitor of formula II. For instance, the present invention includes the use of a glycoprotein processing inhibitor of formula I and a aspartyl protease specific inhibitor of formula II and for the preparation of a pharmaceutical formulation for simultaneous, separate or sequential use for treating an HIV infection wherein the said compounds of formula 1 are ho r2o Formula I OR j. 20 wherein Rj is a Ci_8 alkanoyl, Ci_io alkenoyl, Ci-8 alkoxyacetyl, or benzoyl optionally substituted with an alkyl or halogen group; or a pharmaceutically acceptable salt thereof and said compound of formula II is 25 CNHCH II 0 CNHCHC-CF,C-NR«-R, II II ^1 56 0 0 0 Formula II 30 35 and the hydrates, isosteres and the pharmaceutically acceptable salts thereof wherein Rx is benzyl "^il oxy, S02HN, (3-pyndyl )ethyl, isoquinolyl, 4-alkoxy- benzyloxy, or morpholyl, P2 is isopropyl, cyclopentyl, 2-(4,4-difluoro)-pyrrolidyl, 2-hydroxy-2-propyl, t-butyl, P1 is piperonyl, 4-(benzyloxy)benzyl, 3-(benzyloxy)benzyl, (4-benzyloxy-3-methoxy)benzyl, when R5 is H, R6 is benzyl, WO 94/19008 PCT/US94/00710 -27- piperonyl, CH2~pyridyl, 4-(benzyloxy)benzyl, morpholino, tetrahydroisoquinolyl, 4-(3-hydroxypropyl)benzyl, OH with Y and Z both being as generally defined, but particularly when Y is isopropyl, preferably in the D configuration, or phenyl and when Z is benzyloxymethylene, CHO, COOH, alkoxy or COOR4, when R5 is other than H it is 1° preferred that R5 be methyl, 4-hydroxybutyl or 3-hydroxypropyl and that R6 be benzoxy or benzyl, and when R5 and Rg form a heterocyclic moiety with the nitrogen attached thereto, it is preferred tha -h^ ^ncho and morpholino be the heterocyclic moieties. It is further 5 understood further subgroupings from the above combination may be made from the groups of formula I and II defined herein. A preferred composition containing the compounds of 20 formula I and II are those compositions which contain at least 10% of the compounds of formula I or those compositions that contain at least 10% of the compounds of formula II. 25 The compounds of this invention may be assayed for their inhibition of HIV replication using the following published techniques. The following examples illustrate various aspects of this invention. The following information on reagents, cell lines, virus strains and assays describe the usefulness of the previously described compositions. Further methods and procedures dealing with the assays are known in the art. 5 2-(3-hydroxypropyl)benzyl or -CH(Y)(Z) 30 EXPERIMENTAL PROCEDURES WO 94/19008 PCT/US94/00710 -28- To study the effects of the described compositions the growth of HIV-1 was used to study the treatment of AIDS as a useful model for HIV. 5 Drug Combination Assay For the investigation of drug combinations, the MTT cell viability assay (Pauwels et al., J. Virol. Methods, 1988, 20, 309-321) was used. Various drug combinations were achieved by creating chequerboards with one compound being 10 titrated horizontally with the second compound being titrated vertically across a 96 well microtitre plate using multichannel pipettes. The use of six microtiter plates was required for each assay (only inner 60 wells) with quadruplicate wells for each drug combination. Doubling 15 . dilutions or half log dilutions, with the end rows left drug free, were usually made. MT-4 cells infected with 100 TCIDso of HIV-Irf per 5 x 104 cells were added to each well at a concentration of 5 x 20 104 cells per well and after incubation at 37°C of six days, lOul of acidified isopropanol was added and the plates read at 540nm using a Multiscan MCC/340 spectrophotometer (Flow Laboratories). The raw plate data were captured onto floppy disc using Ultroterm (LKB). Subsequent data reduction was 2 5 performed using Excel (Microsoft). This enabled the mean O.D. values for each drug combination to be calculated and a series of 19 dose response curves, i.e. the dose response of each drug at a fixed concentration of the other to be generated with minimal user intervention. The top four left hand corner wells which received the highest concentration of each drug were used as a positive control (i.e. cells viable totally protected) and the four bottom right hand corner wells without any drug were used as the negative control (i.e. total cell death). AVO 94/19008 PCT/US94/00710 -29- Analysis of Drug Combinations by Isobolic Method From the dose response curves, the irso for each drug, either alone or in combination with a fixed concentration of ^ the other, was calculated and isobolograms plotted (Suhnel, Antiviral Research 13, 23-40(1990)). It was possible to determine from the shape of the isobole whether the drugs had a synergistic relationship (concave), additive effect (linear) or showed anatagonism (convex). Additionally a 10 combination index (CI) was calculated (Suhnel, 1990). 15 20 25 30 The table provides ED50 values computed from dose response lines for each compound in the presence of a fixed concentration of the other compound. From these data was determined the combination index (CI) from the formula:- CI = (D)i + (D)? + a (D)i (D)? (DX)i (DX)2 (Dx)i (Dx)2 where a = 1 for mutually non-exclusive agents and (DX)i = IC 50 of drug 1 above; (DX)2 = IC 50 of drug 2 above; (D)1 and (D)2 = concentrations of drugs 1 and 2 in combination giving 50% inhibition; If it is: <1 (synergism); CI = 1 (zero interaction); >1 (antagonism). (ref: J. Suhnel, Antiviral Research 13 (1990), 23-40.) 35 WO 94/19008 PCT/US94/00710 -30-EXAMPLE 1 ANTIVIRAL ACTIVITY OF RO-31-8959 AND MDL7369 WITH BUCAST 5 Three protease inhibitors MDL73669 (Tables 1A and IB), Ro-31-8959 (Tables 2A and 2B) and MDL 74538 (Tables 3A and 3B) were investigate in combination with the glycoprotein processing inhibitor MDL 28574 (BCJCAST). The method used test the effectiveness of the combination was assessed by 10 looking at the various concentrations of the compounds in combination on antiviral activity determined by using the cell viability assay previously described. The method of study of these compounds allowed for identification of synergistic interactions by analysis of the data values fo 15 a Combination Index (C.I.). The following abbreviations apply to the following compounds: MDL73669 = [1(S)-[[3,3-difluoro-2,4,-dioxo-1-[[4- 20 (phenylmethoxy)phenylJmethyl]-4-[ (phenylmethyl)amino]butyl)aminocarbony1]-2-methylpropyl]carbamic acid, phenylmethyl ester. MDL74538 = N-[4-(N-benzyloxycarbonyl-L-valyl)amino-2,2 di fluoro-1,3,-dioxo-5-(4-benzyloxy)phenyl-pentyl] — (O— 25 benzyl)-D-valinol. MDL28574 = (Bucast) [ IS- (la,68,7a,88,8a8) ]-oct?'.iycUo-1,6,7,8-indolizinetetrol 6-butanoate Ro-31—8959 = N-tertiary butyl—decahydro-2[2(F)-hydroxy 4-phenyl-3(S)-[[N(2-quinolylcarbonyl)-L-30 asparaginyl]amino]-butyl]-(4aS,8aS)-isoquinoline-3(S)- carboxamide. 35 WO 94/19008 PCT/US94/00710 -31- Table 1A and IB: Concentrations of MDL73669 and BMDL28574, in combination and alone required to give 50% protection of MT-4 cells infected with HIV-Irf and the combination indices calculated from these values. 5 10 15 Fixed Cone MDL 73669 (MM) IC 50 MDL 28574 (pM) C.I.* 0 100 - 0.0001 100 - 0.0003 100 - 0.001 45 0.45 0.003 25 0.25 0.010 21 0.22 0.03 32 0.37 0.1 20 0.4 0.3 30 0.95 1 - - *The combination (C.I.) indices indicate synergy Fixed Cone MDL 28574 (pM) IC 50 MDL 73669 (MM) CI.* 0 0.6 - 0-1 0.55 - 0.3 0.6 - 1 0.5 0.85 3 0.5 0.89 10 0.42 0.87 30 0002 0.3 100 - - 300 - - *The combination (C.I.) indices indicate synergy WO 94/19008 PCT/US94/00710 -32- Table 2A and 2B: Concentrations of Ro-31-8959 and MDL 28574, in combination and alone required to give 50% protection of MT-4 cells infected with HIV-Irf and the combination indices calculated from these values. Fixed Cone Ro-31-8959 (nM) IC 50 MDL 28574 (MM) C.I.* 0 100 - 0.1 50 0.52 0.3 60 0.66 1 12 0.28 3 6.8 0.52 10 - - 30 - - 100 - - "The combinationindices (C.I.) indicate synergy I Fixed Cone MDL 28574 (p M) IC 50 Ro-31-8959 (nM) C.I.* 0 6.9 0.1 6.7 - 0.3 9.0 - 1 6.1 - • 3 12 - 10 1.4 0.32 30 0.38 0.37 100 - - 300 - - * I he combinationindices (C.I.) indicate synergy </div>

Claims

<div class="application article clearfix printTableText" id="claims"> ^WO 94/19008 PCT/US94/00710 -33- Table 3A and 3B: Concentrations of 74538 and HDL 28574, in combination and alone required to give 50% protection of MT 4 cells infected with HIV—Irf and the combination indices calculated from these values. Fixed Cone IC 50 MDL 74538 MDL 28574 C.I *;(nM);(MM);0;23;-;0.1;20;0.87;0.3;9.5;0.42;1.0;25;1.1;3.0;8.2;0.43;10.0;6.2;0.39;30.0;3.0;0.48;"The combination (C.I.) indicate synergy.;20;Fixed Cone MDL 28574 (pM);IC 50 MDL 74538 (nM);C.I* 0 100 - 3.0 30 0.47 10.0 2 0.46 25 "The combination (C.I.) indicate synergy. 30 35 -• 261740 WHAT IS CLAIMED IS:

1. A pharmaceutical composition comprising a compound of formula I which is 5 (IS-(la,6b,7a,8b,8ab]-octahydro-1,6,7,8-indolizinetetrol 6-butanoate or a pharmaceutically acceptable salt thereof; and a compound of formula II selected from the group of N-tertiary butyl-decahydro-2[R(R)-hydroxy-4-phenyl-3(S)-[[N(2-quinolylcarbonyl)-L-asparaginyl]amino]-butyl]-(4aS,8aS)-isoquinoline-3(S)-carboxamide; 15 t1(S)-[[3,3-difluoro-2,4,-dioxo-l-I[4-(phenylmethyl)amino]butylJaminocarbony1]-2-methylpropyl]carboxamic acid, phenylmethyl ester; and N-[4-(N-benzyloxycarbonyl-L-valyl)amino-2,2-difluoro-1,3,-dioxo-5-(E-benzyloxy)phenyl-pentyl]-(0-benzyl)-D-valinol; and the hydrates, isosteres and pharmaceutically acceptable salts thereof.

2. A pharmaceutical composition according to claim 1 wherein said compound of formula II is an amount of at least 10% by weight. 30

3. A composition of claim 1, which comprises at least 10% by weight of the compound of formula I.

4. A composition of any preceding claim, adapted for parenteral administration. 261740

5. A composition of any preceding claim, comprising the compound of formula I and the compound of formula II as a physiological functional salt or other derivative thereof, 5 in an acceptable pharmaceutical vehicle.

6. Use of a pharmaceutical composition of any one of claims 1-5 in the manufacture of a medicament of the treatment of an HIV infection. 10

7. A product containing compounds of formula I and formula II as defined in claim 1, as a combined preparation for simultaneous, separate or sequential use in treating an HIV infection.

8. A pharmaceutical composition as defined in claim 1 and the hydrates, isosteres and pharmaceutically acceptable salts thereof substantially as herein described with reference to any example thereof.

9. Use of a pharmaceutical composition as claimed in claim 6 substantially as herein described with reference to any example thereof.

10. A product as claimed in claim 7 substantially as herein described with reference to any example thereof. By the authorised agents A J Per </div>