CA2043143A1 - Retrovirally active new anhydrostatin phosphonopyrrolidines and piperidines - Google Patents

Abstract

ABSTRACT
Retrovirally active new anhydrostatin phosphono-pyrrolidines and piperidines of the formula (I) in which W - is . hydrogen or a typical amino protecting group, or - is straight-chain or branched alkyl or alkenyl in each case having up to 6 carbon atoms, which are optionally substituted by aryl having 6 to 10 carbon atoms, or - is a group of the formula R3-Co-, R5R4N-Co-or R6-SO2-, A, B and D are identical or different and each - represent a direct bond or - represent a radical of the formula , or , or - represent a group of the formula

Description

2~31 ~3 The invention relates to new anhydrostatin pho~phonopyr-rolidines and -piperidines, to a process for their preparation and to their use as medicaments, in parti-cular ~ antiviral agents in human and veterinary medicine.

It has ~lready been attempted to employ p~eudopeptides, which in ~ome cases al~o have rznin-inhibitory ~ctivity, in combating AIDS tcf. GB ~2 203,740; EP 337,714;
EP 342,541; EP 346,847 and EP 352,000].

The present invention relates to new pyrrolidine-~ubsti-tuted and piperidine-substituted pseudopeptides of the genaral formula (I) Rl r (CH2)n W-A-B-D-N~ ( I ) in which 15W - ishydrogen or a typical amino protecting group, or - isstraight-chain or branched alkyl or alkenyl in each ca~e having up to 6 carbon atoms, which are optionally substituted by aryl having 6 to 10 carbon atoms, or ~ is a group of the formula R3-Co-, R R4N-Co-or R~-S02-, Le A 27 644 - 1 -2~314~

in which R3 - denotes hydrogen, trifluoromethyl or straight-chain or branched alkoxy having up to 8 carbon atoms or alkyl having up to 18 carbon atoms, which are optionally substituted by aryl having 6 to 10 carbon atoms or pyridyl, or - denotes aryl having 6 to 10 carbon atoms, which is optionally substituted by halogen, tri-fluoromethyl, trifluoromethoxy or by straight-chain or branched alkyl having up to 8 carbon atoms, - denotes cycloalkyl having 3 to 7 carbon atoms, - denotes indolyl, pyridyl, morpholino or piper-azinyl, or - denotes a radical of the formula ~R7 ~7 ~R7 R8-Y-CH2-CH-, R9-Co-o-CH-, R10-S(O)m-NH-CH-, ~R7 T-NH-tCH2)p~ 0~ N-Y'-(CH2)~-CH-~

0 ~7 R~ 1 -p- ( CH2 ) s-CH- or R7(CH2)2-CH-R 1 1 ~
in which R7 - denotes phenyl or naphthyl, Le A 27 644 - 2 -2~3~3 R8, R9 and R10 are identical or different and - denote straight-chain or branched alkyl having up to 17 carbon atoms, which i~
optionally substituted by phenyl or naphthyl, or denote aryl having 6 to 10 carbon atoms, which is optionally substituted by alkyl having up to 4 carbon atoms, m - denotes a number 0, 1 or 2, ~ - denotes morpholino or cyclohexyl, p - denotes a number 1, 2 or 3, Y and Y' are identical or different and denote the CO or SO2 group, t - denotes a number 0 or 1, Rl1 and R11 are identical or different and denote hydroxyl or alkoxy having up to 8 carbon atoms, and s - denotes a number 1 or 2, R4 and R5 are identical or different and - denote hydrogen or - denote aryl having 6 to 10 carbon atoms, which i8 optionally substituted by straight-chain or branched alkyl having up to 6 carbon atoms or halogen, or - denote cycloalkyl hav~ng 3 to 7 carbon atoms, or - denote straight-chain or branched alkyl having up to 18 carbon atoms, which is optionally substituted by pyridyl, R~ - denotes straight-chain or branched alkyl having up to 8 carbon atoms, Le A 27 644 - 3 -A, B and D are iden~ical or different and each - represent a direct bond or - represent a radical of the formula (H C) 2 Ix ~ ~3C~H3 ~N ~ CO- , -N ~ tCH2)r CO

(CH2~
rl S~s or -NH~^~CO , in which x and x' are identical or different and are the number 1 or 2 and r - denotes the number 0 or 1, or - represent a group of the formula _NR12 1 ~CH2)z-C~-in which z - denotes the number 0 or 1, R12 denotes hydrogen or ~traight chain or branched alkyl having up to 8 carbon atoms, Rl3 denotes cycloalkyl having 3 to 8 carbon atoms or aryl having 6 to 10 carbon atoms or hydro-gen, or - denote~ straight-chain or branched alkyl having he A 27 644 - 4 -~_ . _ 2~ 73~ ~ ~

up to 8 carbon atoms, which is optionally substituted by alkylthio having up to 6 carbon atoms, hydroxyl, mercapto, guanidyl or by a group of the formula -NRl4R'5 or R16-OC-, S in which R14 and R15 are identical or different and denote hydrogen, straight-chain or branched alkyl having up to 8 carbon atoms or phenyl, and R1~ denotes hydroxyl, benzyloxy, alkoxy having up to 6 carbon atoms or the abovementioned group -NR14R15, or which is optionally substituted by cyclo-alkyl having 3 to 8 carbon atoms or by aryl ha~ing 6 to 10 carbon atoms, which is optionally substituted by hydroxyl, halogen, nitro, alkoxy having up to 8 carbon atoms or by the group in which R14 and R15 have the abovementioned meanin~, or which is optionally substituted by a 5- or 6-membered nitrogen-containing heterocycle or indolyl, in which the corresponding -NH func-tions are optionally protected by alkyl having up to 6 carbon atoms or by an amino protecting group, R1 - is cycloalXyl having 3 to 8 carbon atom~, - is straight-chain or branched alkyl or alkenyl having up to 10 carbon atoms, which are Le A 27 644 - 5 -~J~ 3 ~

optionally substituted by cycloalkyl having 3 to 8 carbon atom~ ox aryl h~ving 6 to 10 carbon atoms, which is op~oxally substituted by halogen, nitro, hydroxyl, amino or straight-chain or branched alkoxy having up to 6 carbon atoms, n - the number 1 or 2, R2 _ a radical of the formula O=PRl lRl l , -co-Rl7 or -co-NR4R5, in which R4, R5, Rll and Rll have the abovementioned m anings, and Rl7 - denotes straight-chain or branched lkoxy having up to 8 carbon atoms, which is optio~-ally substituted by phenyl, and their physiologically acceptable salts.

The compounds of the general formula (I) according to the invention ha~e several asymmetric carbon atoms. They can he present independently of one anothex in ~he D- or the L~ form. The invention includes the optical antipodes as well as the isomer mixtuxes or racemates. Preferably, the groups A, B and D ar~ present independently of one another in the optically pure form, preferably in the L-foxm.

The radical of the general formula (VIII) Le A 27 644 - 6 -~. l3~ t'~L~3 R~ (CH2)~
-NH ~ ~ (VITI

has 2 asymmetric carbon atoms (1,2), which can be present independently of one another in the R- or S- config~ra-tion.

The geometry of the double bond can be both trans ~E) as well as cis t~) Amino protecting groups in the context of the invention are the amino protecting groups cu3tomary in peptide chemistry.
These preferably include: benzyloxycarbonyl, 3,4-dLmeth-oxybenæyloxycarbonyl, 3,5-dimethoxybenzyloxycarbonyl, 2,4~dimethoxybenzyloxycarbonyll 4-methoxybenzyloxy-carbonyl, 4-nitrobenzyloxycarbonyl, 2 nitrobenzylo~y-carbonyl, 2-nitxo-4,5-dimethoxybenzylo~ycarbonyl, meth-oxycarbonyl, ethoxycarbonyl, propo~ycarbonyl, i~opropoxy-carbonyl, butoxycarbonyll i~obutoxycarbonyl, tert-butoxy-carbonyl, 2-nitrobenzyloxycarbonyl, 3,4,5-trimethoxy-benzyloxycarbonyl, methoxycarbonyl~ ethoxycarbonyl, propoxycarbonyl, i~opropoxycarbonyl, butoxycarbonyl, isobutoxycarbonyl, tert-butoxycarbonyl, cyclohexoxy-carbonyl, 1,l-d~ethylethoxycarbonyl, adamantylcarbonyl, phthaloyl, 2,2,2-trichloroethoxycarbonyl, 2,2,2-tri-chloro-tert-butoxycarbonyl, menthyloxycarbonyl, phenoxy-carbonyl, 4 nitropheno~yearbonyl, fluorenyl-9-methoxy-Le A 27 644 - 7 -2~ 3 carbonyl, formyl, acetyl, propionyl, pivaloyl, 2-chloro-acetyl, 2-bromoacetyl, 2,2,2-trifluoroacetyl, 2,2,2-trichloroacetyl, benzoyl, 4-chlorobenzoyl, 4-bromo-benzoyl, 4-nitrobenzoyl, phthalimido, isovaleroyl or benzyloxymethylene.

The compounds of the general formula (I) according to the invention can be present in the form of their salts.
These can be salts with inorganic or organic acids or bases.

Preferred compounds of the general formula (I) are those in which W - is hydrogen, tert-butyloxycarbonyl ~BOC), 9-fluorenylmethyloxycarbonyl (Fmoc) or benzyloxy-carbonyl (Z) or ~ is straight-chain or branched alkyl or alkenyl in each case having up to 4 carbon atoms, which are optionally substituted by phenyl, or - is a group of the formula R3-Co- or R5R4N-co-, in which R3 - denotes hydrogen, trifluoromethyl or straight-chain or branched alkoxy having up to 4 carbon atom~ or alkyl having up to 16 carbon atoms, which are optionally substituted by phenyl, naphthyl or pyridyl, or - denotes phenyl or naphthyl, each of which is optionally substituted by fluorine, chlorine, t~ifluoromethyl, trifluoromethoxy or by Le A 27 644 - 8 -~.31 ~3 ~traight-chain or branched alkyl having up to 6 carbon atoms, - denotes cyclopropyl, cyclopentyl, cyclohexyl, indolyl, pyridyl, morpholino or piperazinyl~ or - denotes a radical of the formula ~ 7 ~ 7 R8-Y-CH2-CH-, R9-Co-o-CH-~ 7 Rl-5tO)m-NH-CH-in wh~ch Y - denotes the CO or SOz group, R7 - denotes phenyl or naphthyl, RB, R~ and R10 are identical or different and denote straight-chain or branched alkyl having up to 15 carbon atoms, tolyl, phenyl or naphthyl, and m - denotes a number 1 or 2, R4 and R5 are identical or different and - denote hydrogen or - denote phenyl or naphthyl, each of which is optionally substituted by ~traight-chain or branched alkyl having up to 4 carbon atoms, fluorine or chlorine, - denote cyclopropyl, cyclopentyl or cyclohexyl, or - denote ~traight-chain or branched alkyl having up to 16 carbon atoms, which i~ optionally sub3tituted by pyridyl, Le A 27 644 - 9 -2~31~3 A, B and D are identical or different and - represent a direct bond or - represent proline, or - represent a radical of the formula r(lH2)X' S~ ~S
S H~C32 ) -CO- -N}~ ~CO

in which r denotes the number O or 1, and x' denotes the number 1 or 2, - represent a group of the formula -NR~2 1 ICH2)z-CO-in which z - denotes the number O or 1, R12 - denotes hydrogen or straight-chain or branched alkyl having up to 6 carbon atoms, Rl3 - denotes cyclopentyl, cyclohexyl, phenyl or hydrogen, - or denotes 6traight-chain or branched alkyl having up to 6 carbon atoms, which can option-ally be ~ubstituted by hydroxyl, carboxyl or H2N-CO- ~
or i8 substituted by cyclohexyl, naphthyl or phenyl, each of which is optionally Le A 27 644 - 10 -2~d-31~3 substituted by fluorine, hydroxyl, nitro or alkoxy having up to 4 carbon atoms, or is substi~uted by indolyl, imidazolyl, pyridyl, triazolyl or pyrazolyl, where the corresponding -2~H
functions are optionally protected by alkyl having up to 4 carbon atoms or by an amino protecting group, Rl - represents cyclopropyl, cyclopentyl or cyclohexyl, - represent~ straight-chain or branched alkyl or alkenyl having up to 8 carbon atoms, which are optionally substituted by cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl or phenyl, each of which is optionallysUbstituted by fluorine, chlor-ine, bromine, nitro, hydroxyl or amino, n - represents the number 1 or 2, and R2 _ represents a radical of the formula ~=p_RllRll , _CC_R17 or -Co-NR4R5 in which R4 and R5 are identical or different and have the abovementioned meaning~
Rll and Rll are identical or different and denote hydroxyl or straight-chain or branched alkoxy having up to 6 carbon atoms, and Rl7 - denotes straight-chain or branched alkoxy in each case having up to 6 carbon atoms, which is optionally substituted by phenyl, and their physiologically acceptable ~alts.

Le A 27 644 - 11 -2 ~ 3 Particularly preferred compounds of the general formula (I) are those in which W - represents hydrogen, tert-butyloxycarbonyl (BOC) or benzyloxycarbonyl (z)~ or - represents allyl or benzyl, - represents a group of the formula R3-Co- or R5R4N-co-, in which R3 - denotes hydrogen, trifluoromethyl or straight-chain or branched alkyl having up to 14 carbon atoms which are optionally substituted by phenyl, naphthyl or pyridyl, or - denotes phenyl or naphthyl, each of which is optionally substituted by fluorine, chlorine, trifluoromethyl, trifluoromethoxy or straight-chain or branched alkyl having up to 4 carbon atoms, - denotes cyclopropyl, cyclopentyl, cyclohexyl, indolyl, pyridyl, morpholino or piperazinyl, or - denotes a radical of the formula or ~R
Rl-S(O)m-NH-CH-, in which Y - denotes the CO or SO2 group, R7 - denotes phenyl or naphthyl, R~, R~ and R10 are identical or different and Le A ?7 644 - 12 -2 ~

denote ~traight-chain or branched alkyl having up to 14 carbon atoms, tolyl, phenyl or naphthyl, and m - denotes a number 1 or 2, R4 and Rs are identical or different and - denote hydrogen or - denote phenyl or naphthyl, each of which i~
optionally substituted by methyl, fluorine or chlorine, - denote cyclopropyl, cyclopentyl or cyclohexyl, or - denote etraight-chain or branched alkyl having up to 14 carbon atoms, which is optionally substituted by pyridyl, A, ~ and D are identicsl or differen~ and - represent a direct bond, or - represent proline, or - represent a group of the formula R ~

-NR1~A~CH2)z-Co , in which z - denotes the number O or 1, RlZ _ denotes hydrogen or methyl, Rl3 - denotes cyclopentyl or cyclohexyl, - or denotes straight-chain or branched alkyl having up to 4 carbon atoms, which is option-ally ~ub~tituted by hydroxyl, carboxyl or ~zN-CO-, or i~ substituted by cyclohexyl, naphthyl or Le A 27 644 - 13 -5 ~'3 phenyl, each of which can in turn be substitu-ted by fluorine, chlorine or alkoxy having up to 4 carbon a~Qms~ or is substituted by imidazolyl, triazolyl~
pyridyl or pyrazolyl, where the NH function is optionally protected by methyl, benzyloxymethy-lene or t-butyloxycarbonyl ( BGC ), Rl - represents cyclopentyl or cyclohe~yl, or - represents straight-chain or branched alkyl havinq up to 6 carbon atoms, which i6 optionally substitu-ted by cyclopropyl, cyclopentyl, cyclohPxyl or phenyl, each of which is optionally substituted by hydroxyl, n - reprasents the number 1 or 2 r and RZ _ represents a radical of the formula O=pRllRll , -Co-R17 or -CC-NR4R5 in which R4 and R5 are identical or diff~rent and have the abovementioned meanings R11 and R11 are identical or different and denote hydroxyl or ~traight-chain or branched alkoxy having up to 4 carbon atoms, and Rl7 - denotes straight-chain or branched alkoxy in each ca~e having up to 4 carbon atoms, which is optionally substituted by phenyl, and their physiologically acceptable salts.

A process for the preparation of the compounds of the Le A 27 644 - 14 -2~3~ ~3 general formula ~I) according to the invention R1 r ~CH2)~
W-A-B-D-O R
in which W, A, B, D, Rl, RZ and n have the abovementioned meaning~
has additionally been found, characteriz-ed in that [A] compounds of the general formula tIa) Rl ~CH2)n H2 ~ (Ia) O R
in which Rl, R2 and n have the abovementioned meaning~
are condensed via the corresponding salts, prefer-ably via the trifluoroacetates, either with compounds of the general formula (II) W-A-B-D-OH (II) in which W, A, B and D have the abovementioned meaning~
with acti~ation of the carboxylic acid, or with compounds of the general formula (III) or ( IV) W-X (III) (W'')20 (IV) Le A 27 644 - 15 -J/ ~ 3 in which W has the abovementioned meaning, X represents halogen, preferably chlorine, and 5W~ represents the group CF3CO or CH3CO, under conditions customary in peptide chemistry, in inert solvents, if appropriate in the pre~ence of an ~uxiliary, or [B] compounds of the general formula (v) or (VI) W ' - NH~DHt~3 ~?1 ~-A-B-D-NH ~ H (VI) in which R1, W, A, B and D have the abovementioned meanings and W' represents an amino protecting group, preferably BOC, are condensed with compounds of the general formula (VII) r~ CH2)n ~ (VII) Le A 27 644 - 16 2~3~ ~

in which R2 and n have the abovementioned meaningfi with activation of the carboxylic acids, if approp-riate in the presence of a base and of an auxiliary, and in the case of the compounds of the general formula (V), the protecting group W' is then remuved by a customary method and, if appropriate, reacted further with the compounds of the general formula (II), (III) or (IV) by the method described under process [A].

The proces~ according to the invention can be illustrated by the following equatio~
[A]

2 x CF3COOH x NH2 ~ ~ H3c ~ o-C
o =P(oc2Hs)2 ~O
H3C~O-N~ f~
O O=P ( OC2H5 ) 2 Le A 27 644 - 17 -20~3~3 . . ~B]

,~ Ho~T, DCC
BOC ~ NH H
o o=P(OC2H5)2 - BOC
~OC-N ~ tA~ I BOC-Ser-Ph--A-n-OH
o o~P(OC2H5)2 ~O

~c-S-r -Ph--A-n-N ~
o O~P~oc2H5)2 Suitable solvents for all process steps are the customary inert solvents which do not change under the reaction ; 5 conditions. These preferably include organic solvents such as ethers, for example diethyl ether, glycol mono-methyl ether or glycol dimethyl ether, dioxane or tetra-hydrofuran, or hydrocarbon~ such as benzene, toluene, xylene, cyclohexane or mineral oil fractions or halogeno-hydrocarbon~ such as methylene chloride, chloroform, c~rbon tetrachloride, or dimethyl sulphoxide, dimethyl-formamide, hexamethylphosphoric tri~mide, ethyl acetate, pyridine, triethylamine or picolines. It is also possible to use mixture~ of the solvents mentioned.

Le A 27 644 - 18 -.2~ ~ 3~3 Dichloromethane, chloroform, dimethylformamide or tetra-hydrofuran are particularly preferred.

Auxiliarie~ employed for the respective peptide couplings are preferably conden~ing agents which can also be bases, in particular if the carboxyl group is activated as the anhydride. The customary condensing agents are preferred here, such as carbodiimides, for example N,N~-diethyl-, N,N~-dipropyl-, N,N~-diisopropyl- and N,N~-dicyclohexyl-carbodiimide, N-(3-dLmethylaminoisopropyl)-N'-ethyl-carbodiimide hydrochloride, N-cyclohexyl-N'-(2-morpho-linoethyl)-carbodiimide metho-p-toluenesulphonate, or carbonyl compounds such as carbonyldiimidazole, or 1,2-oxazolium compounds such as 2-ethyl-5-phenyl-1,2-oxazolium-3-sulphate or 2-tert-butyl-5-methyl-isoxazolium perchlorate, or acylamino compounds such as 2-ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline, or propanephosphonic anhydride, or isobutyl chloroformate, or benzotriazolyloxy-tris(dimethylamino)phosphonium hexafluoropho~phate or 1-hydroxybenzotriazole.

Additionally, for example, alkali metal carbonates, for example sodium carbonate or potassium carbonate or sodium hydrogencarbonate or potassium hydrogencarbonate, or organic bases ~uch as trialkylamines, for example tri-ethylamine, N-ethylmorpholine, N-methylpiperidine or N-methylmorpholine can be employed. N-Nethylmorpholine is preferred.

The auxiliaries and bases are employed in an amount of Le A 27 644 - 19 -" 2~3~

1.O mol to 3.0 mol~ preferably 1.O to 1.2 mo~, in each case relative to 1 mol of the compounds of the general formula (V) or (VI).

The peptide couplings are carried out in a temperature range from 0C to 100C, preferably at 0C to 30C and at normal pressure.

The reactions can be carried out both at normal pressure and at elevated or reduced pressure (for example 0.S to 5 bar), preferably at normal pressure.

The compounds of the general formula (Ia) are also new and can be prepared by the abovementioned process ~B].

The compounds of the general formulae (II~, (III) and (IV) are known or can be prepared by a customary method.

The compounds of the general formulae (V) and (VI) are known in some cases or are new and in the latter case can be prepared starting from the corresponding esters by hydrolysis according to a customary method tcf.
US 4,725,584; EP 209,897; EP 163,237; J. Med. Chem. 31, 1377 (1988), 29, 104 (1986); THL, 43, 4297 (1987)].

The compounds of the general formula (VII) are also known ~n=l cf. US 4,186,268; Y. Nomura et al., Chem. Lett., 693 (1977); n = 2 cf. V.A. Solodenko et al., Zh. Obshch.
Rhim. S7, 2392 (1987)].

Le A 27 644 - 20 -It has surprisingly been found that the compounds of the general formula (I) have an extremely strong action against retroviruses. This is confirmed using an HIV-speci~ic protease enzyme test.

The results for the examples listed below were determined by the HIV test sy~tem described in the following litera-ture reports [cf. Hansen, J., Billich, S., Schulze, T., Sukrow, S. and Molling, R. (1388), EMB0 Journal, Vol. 7, No. 6, pages 1785-1791~: purified HIV protease was incubated with synthetic peptide which imitated a cleav-age site in the Gag precursor protein and representPd an in vivo cleavage site of the HIV protease. The resulting cleavage products of the synthetic peptide were analyzed by means of rever e phase hi~h performance liquid chroma-tography (RP-HPLC). The IC50 values given relate to the substance concentration which causes a 50 ~ inhibition of protea~e activity under the abovementioned test condi-~ions.

Table I:
Ex2mple No. IC50 (RP-HPLC~ ~M~
_ 3 ~ 10 5 7 < ~0 5 27 < 10-5 33 10-~
< 10-5 62 ~ 10 5 Le A 2? 644 - 21 -2~31~^3 The compounds according to the invention additionally showed action in infected cell culture. It was possible to show this by the example of the visna viru~ and HIV
virus tcf. ~uhnel, H. et al., Proc. Natl. Acad. Sci., USA
Vol. 86, p. 2382 (1989); Popovic, M. et al., Science 224, p. 497 (1984)].

The visna virus and the HIV virus (human immunodeficiency virus) bo~h belong to the retrovirus subfamily of the lentiviruses. Both viruses have a similar genome organiz-ation and a complex transcription pattern compared to the other retroviruses.

Rnown inhibitors of HIV also inhibit the visna virus in vitro in comparable concentrations, i.e. this model is suitable for the testing and discovery of inhibitors of HIV.

In cell cultures which are infected with visna virus, pronounced virus-induced cytopathic effects occur 5 to 10 days after infection. It was possible to prevent the occurrence of these cytopathic effect~ by treatment of the infected cell cultures with the co~pounds according to the invention.

The visna viru~ test was carried out according to the method of 0. Narayan et al., Journal of Infectious Diseases 135, 5, 1977, 800 - 806. To this end, the compounds according to the invention were diluted in culture medium to non-cytotoxic concentration~ in 96-well Le A 27 644 - 22 -~ 7~

microtitre plates. Sheep fibroblast cells (5 x 104 cells per well) were then added to each well in production medium. Each well then contained 50 ~1 of a visna viru~
solution having a titre of about 2.5 x 104 TCID~o (TCID =
tissue culture infectious dose). This virus dose corres-ponds to an ~OI (multiplicity of infection) of about 0.05.

Under these infection conditions, a virus-induced cyto-pathic effect resulted be~ween day 5 and day 10 in one infection control without substance. The infected and treated cells and the control cells were incubated at 37C in 5 ~ CO2 for 7 days.

When the virus-induced cytopathogenic effect occurred in the untreated virus con~rol, the cultures were fixed with formalin and then stained u~ing a Giemsa solution. The inhibi~ory concentration (IC50) was determined by micro-scopy as the concentration at which the cytopathic effect was inhibited by 50 ~ in comparison ~o the untreated virus control, which showed 100 % cell destruction.

It was found that the compound6 according to the inven-tion protect cells infected with visna virus from virus-induced cell destruction.

Le A 27 644 - 23 -2 & ~

Table II-Example No. IC50 (~M) _ _ 46 (non-polar isomer) 4.1 46 (polar isomer) 6.1 61 5.2 62 48.9 The compounds according to the invention are suitabls as active compounds in human and veterinary medicine for the treatment and prophylaxis of disease caused by retro-viruses.

Examples of indication areas which can be mentioned in human medicine are:
1.) The treatment or prophylaxis of human retrovirus infections.
2.) For the treatment or prophylaxis of diseases (AIDS) caused by HIV I (human immunodeficiency virus;
earlier called HTLV III/LAV) and by HIV II and the stages associated therewith such as ARC (AIDS-related complex) and LAS (lymphadenopathy syndrome) and al80 tha immunodeficiency and encephalopathy caused by this virus.
3.) Por the treatment or the prophylaxis of an HTLV I or HTLV II infection.

4.) For the treatment or the prophylaxis of the ~e A 27 644 - 24 -HIV-carrier state (HIV-transmitter state).
Examples of indications in veterinary medicine which can be mentioned are:

Infections with a) Maedi-visna (in sheep and goats) b) progressive pneumonia virus (PPV) (in sheep and goats ) c) caprine arthriti6 encephalitis viru~ (in ~h~ep and goats~
d) Zwoegerziekte ~irus (in shPep) e) infectious anaemia virus (of the horse) f) inf~ctions caused by the feline leukaemia Yirus.
g) infection~ caused by the feline Lmmunodeficiency virus.

The abovementioned items 2, 3 and 4 are preferred from the indication area in human medicine.

The present invention includes pharmaceutical prepara-tions which contain one or more compound~ of the formula (I) or which consist of one or more active compounds of the formula (I) in addition to non-toxic, inert pharma-ceutically suil:able excipients, and processes for the production of these preparations.

The active compounds of the formula (I) are intended to be present in the abovementioned pharmaceutical prepara-tions, preferably in a concentratlon of about 0.1 to 99.5, prefer~bly from about 0.5 ~o 95% by weight, of the Le A 27 644 - 25 -2~3~ ~3 total mixture.

The abovementioned pharmaceutical preparations may also contain other pharmaceutical active compounds in addition to the compounds of the formula (I).

The abovementioned pharmaceutical preparations are prepared in a customa~y manner by known methods, for example by mixing the active compound or compounds with the excipient or excipients.

In general, it ha~ proved advantageous both in human and in veterinary medicine to administer the active compound or compounds of the formula (I) in total amounts from about 0.5 to about 500, preferably 5 to 100 mg/kg of body weight every 24 hours, if desired in the form of several individual doses, in order to achieve the desired results. An individual dose contains the active compound or compounds preferably in amounts from about 1 to about 80, in particular 1 to 30 mg/kg of body weight. However, it may be necessary to deviate from the dosages men-tioned, in particular depending on the species and the body weight of the sub~ect to be treated, the type and severity of the disQase, the type of preparation and the administration of the medicament (e.g. oral1y or by in~iection) and the period or interval within which administration takes place.
.

Le A 27 644 - 26 -2~ ~31~

Appendix to the experimental section I. List of the eluent mixtures used for chromatoaraphy-I Dichloromethane : methanol II Toluene : ethyl acetate III Acetonitrile : water IV Dichloromethane : methanol : ammonia 9:1:0.1 II. Amino acids In general, the configuration is indicated by placing an L or D before the amino acid abbreviation, in the case of the racemate a D,L, it being possible, for simplifica-tion, to suppress the indication of configuration in the case of L-amino acids and explicit indication then only taking place in the case of the D-form or of the D,L-mixture.

Ala L-alanine Arg L-arginine Asn L-asparagine Asp L-aspartic acid Cys L-cysteine Gln L-glutamine Glu L-glutamic acid Gly L-glycine His L-hi~tidine Ile L-isoleucine Leu L-leucine Lys L-lysine Le A 27 644 - 27 -2~31~3 Met L-methionine Pro L-proline Phe L-phenylalanine Ser L-serine Thr L-threonine Trp L-tryptophan Tyr L-tyrosine Val L-valine III. Abbreviations BOC tert-butoxycarbonyl CMCT l-cyclohexyl-3-(2-morpholinoethyl)-carbodiimide metho-p-toluenesulphonate DCC dicyclohexylcarbodiimide DMF dimethylformamide HOBT l-hydroxybenzotriazole Mir miristoyl Ph phenyl THF tetrahydrofuran Starting compounds Example I

(4S)-4-t(tert-Butoxycarbonyl)amino]-5-phenyl-2-pentenoic acid ~h BOC -NH~OOH

Le A 27 644 - 28 -~ t~ '7' .? ~ C~

9.26 g (30.3 mmol) of me~hyl (4S)-4-[(tert-butoxy-carbonyl)amino]-5-phenyl-2-pentenoate [cf. D.H.R. Barton et al~, ~etrahedron 43, 4297 (1987); S.A. Thompson et al., J. Med. Chem. 29, 104 (1986)~ and 2.54 g (60.6 mmQl) of lith~um hydroxide hydrate were heated to reflux in a mixture of lfi ml of tetrahydrofuran and 7 ml of water for 15 minutes. The mixture was then stirred into a mixture of 200 ml of ethyl acetate and 200 ml of water. The aqueous phase (about pH 10) was separa~ed off, 100 g o~
ice and 2Q0 ml of ethyl acetate were added and the mixture was adjusted to pH 3.0 using lN hydrochloric acid. The organic pha~e was separa~ed off and the aqueous phase was extracted again with 50 ml of ethyl acetate.
The combined organic extr~cts were dried o~er magnesium sulphate. After evaporating the solvent and triturating ~he residue with a little diethyl ether, 7.07 g ~30 ~ of theory) of the title compound were obtained as cOlorless crystals.
Melting point: 160 C
R~ = 0.59 (acetonitrile : water = 9:1) MS (FAB): m/e = 292 (M+H)+, 314 (M+Na)+, 335 (M+2Na-H)+

Example II
(4S~-4-[(tert-Butoxycarbonyl)amino]-5-cyclohexyl-2-pentenoic acid ,~J
soC- ~ ~COOH

Le A 27 644 - 29 -~3~

As deficribed for Example I, 123 g (81 ~ of theory) of the title compound were obtained as colo~less crystals from 159 g (511 mmol~ of methyl (4S)-4-[(tert-butoxycarbonyl)-amino]-5-cyclohexy1-2-pentenoate [J.R. Luly et al., US 4,725,584].
Melting point: 140C
Rf = 0.34 (dichloromethane : methanol = 95:5) MS (DCI, NH3): m/e = 298 (M+H)+, 315 (M+NH4)+

Example III
N-(tert-Butoxycarbonyl)-N-[2-~2-pyridinyl)ethyl]-L-prolinamide rl BOC- ~ ~
CO~

9.2 ml (77.0 mmol) of 2-(2-aminoethyl)-pyridine, 13.4 g (87.5 mmol) of 1-hydroxybenzotriazole (HOBT) and 16.61 g (80.5 mmol) of dicyclohexylcarbodiimide (DCC) were added at 0C to a stirred solution of 15.06 g (70.0 mmol) of BOC-L-proline in 100 ml of dichloromethane. The mixture was then stirred at room temperature for 15 h, the precipitated urea was removed by filtration and the filtrate wa~ concentrated in vacuo. The residue was dis~olved in 200 ml of ethyl acetate, washed with satur-ated sodium bicarbonate solution and dried over sodium sulphate. After chromatography of the crude product on silica gel ~dichloromethane : methanol = 10:1), 20.6 g Le A 27 644 - 30 -`, ,7 rA . ,~ "

(92 ~ of theory) of the title compound were obtained.
R~ = 0.50 (dichloromethane ~ methanol = 10:1) MS (EI, 70 eV): m/e = 319 ~M)+

Example IV
N-L2-(2-Pyridinylethyl)~-L-prolinamid~

~CO~

A solution of 20.6 g (65 mmol) of the compound from Example III in 150 ml of a 4N solution of gaseous hydrogen chloride in dioxane was stirred at 0C in the presencP of 30 ml of anhydrous methanol for 8 h. The mixture was then concentrated in vacuo and the residue was dissolYed in 10 % strength aqueous sodium carbonate solution (pH 10).
This ~olution was concentrated to d~ynes~ and the residue was extracted using warm dichloromethane. After drying the organic pha~e over sodium sulphate and evaporating the solvent in vacuo, 12.4 g (87 % of theory~ of the title compound were obtained.
Rf = O.34 (dichloromethane:methanol:ammonia = 3:1:0~1) Example V
Mir-Phe-Phe-Val-OCH3 Le A 27 644 - 31 2~3~3 ~Ph O
H ll CH3(CH2)12~ ~ ~COOCH3 O ~P~
2.17 g (10.5 mmol) of DCC were added to a stirred solution, cooled to 0C, of 3.94 g (10.5 mmol) of N-miristoyl-L-phenylalanine tcf. US 4,396,543;
GB 2,120,257; A.V. Prabhudesai et al., Chem. Phys. Lipids 22, 71 (1978)] and 1.49 g (11.0 mmol) of HOBt in 30 ml of anhydrou~ dimethylformamide and the mixture wa~ stirred at 0C for 2 h. A solution of 3.14 g (10.0 mmol) of HCl-Phe-Val-OCH3and 3.30 ml (30.0 mmol) of N-methylmorpholine in 15 ml of dimethylformamide was added dropwise to this mixture and it was then stirred in a thawing ice bath for 15 h. The resulting precipitate was removed by filtration and washed with 50 ml of toluene. The filtrate was concentrated in vacuo, 50 ml of toluene were added and the mixture was concentrated again. The residue was dissolved in 200 ml of ethyl acetate, a little insoluble material was removed by filtration, and the organic phase was washed with 100 ml of water and dried over magne~ium sulphate. After evaporating the solvent in vacuo and chromatography of the residue on 350 g of silica gel (toluene : ethyl acetate 1:1), 1.9 g (30 % of theory) of the title compound were obtained as a Colorl~ss powder.
Melting point~ 140 DC
R~ 5 0.42 (toluene ~ ethyl acetate 1:1) MS (FAB) m/e = 636 (M+~)+, 658 (M+Na)+

Le A 27 644 - 32 -2 ~ 4 ~

Example VI
Mir-Phe-Phe-Val-OH

O
~ H ~ ~ ~
CH3(CH2)12C ~ H OOH
O ~Ph A solution of 170 mg (4.0 mmol) of lithium hydroxide hydrate in 4 ml of water was added to a solution of 1.27 g (20 mmol) of the compound from Example V in 15 ml of tetrahydrofuran and the mixture was 6tirred at room temperature for 3 h. The reaction mixture was then poured into a mixture of 50 ml of water, 10 g of ice and 50 ml of ethyl acetate and adjusted to pH 3 by addition of lN
hydrochloric acid. The organic phase was separated off, the aqueous phase was extracted with 50 ml of ethyl acetate and the combined organic extracts were dried over magnesium sulphate. After evaporating the solvent in vacuo and treating the residue with 5 ml of ether and 30 ml of n-pentane, 1.03 g (83 % of theory~ of the title compound were obtained as Colo~less crystals.
Melting point: 173-C
HPLC purity: > 96 %
Rs - 0.41 (acetonitrile : water = 9:1) MS (FAB): m/e 622 (M+H)~, 644 (M+Na)~

Le A ?7 644 - 33 -2~31~

Example VII
Mir-Val-Phe-Val-OCH3 As described for Example V, 1.70 g (29 % of theory) of the title compound were obtained as cOlOrles~ crystals from 3.44 g (10.5 mmol) of N-miristoyl-L-valine lcf.
V. Iyer et al., J. Indian Chem. Soc. 59, 856 (1982);
DE 2,234,399; FR 2,192,79S] and 3.14 g (10.0 mmol) of HCl x Phe-Val-OCH3.
Melting point: 170C
R~ = 0.36 (toluene : ethyl acetate 1:1) MS (FAB): m/e = 588 (N+H)~, 600 (M+Na)t Example VIII
Mir-Val-Phe-Val-OH

As described for Example VI, 780 mg (79 % of theory) of the title compound were obtained as colorless crystals from 1.06 g (1.8 mmol) of Mir-Val-Phe-Val-OCH3.
Melting point: 233C
HPLC purity: ~ 94 %
R~ = 0.36 (acetonitrile : water 9:1) MS (FAB): m/e = 580 (M+Li)'; 596 (M+Na)t Le A 27 644 - 34 -2 ~ 3 Pre~aration Examples (general formula I) Example l Diethyl 1-{(4S)-4-[(tert-butoxycarbonyl)amino]-5-phenyl-2-pentenoyl}-(2R,S)-2-(pyrrolidinyl)-phosphonate ~ I
BO~ - N
H
o O=P ( oC2H5 ) 2 3.1S g (23.32 mmol) of HOB~ and 4.59 g (22.3 mmol) of DCC
were added at 0C to a solution of 6.18 g (21.2 mmol) of the compound from Example I in 50 ml of anhydrous dimethylformamide. The cooling bath was removed and the mixture was then stirred at room temperature for 1 h. It was then cooled again to 0C and a solution of 4.83 g (23.32 mmol) of diethyl (2R,S)-2-(pyrrolidinyl)-phosphon-ate [cf. E.W. Petrillo et al., Tetrahedron Lett. 51, 4929 (1979); US 4,186,268] in 50 ml of dimethylformamide and 5.1 ml (46.64 mmol) of N-methylmorpholine was added dropwise. The cooling bath was removed and the mixture was stirred at room temperature for 2 h. ~he resulting precipitate wa~ removed by filtration, 50 ml of toluene were added to the filtrate and the mixture was concen-trated in vacuo. The residue was dissolved in 100 ml of ethyl acetate, washed with saturated sodium bicarbonate solution and saturated sodium chloride solution and dried over magnesium sulphate. After evaporating the solvent in vacuo and chromatography of the crude product on 600 g of silica gel (dichloromethane : methanol = 95:5), 8.10 g (80 % of theory) of the title compound were obtained as Le A 2? 644 _ 35 _ - a viscous oil.
Rr = 0.29 (dichloromethane : methanol 95:5) MS (FAB) m/e = 481 (M~H)~, 503 ~M+Na)t Example 2 1-{(4S)-4-[(tert-butoxycarbonyl)amino]-5-phenyl-2-penten-oyl}-N-[2-(2-pyridinylethyl)]-L-prolinamide ~h I--BOC -~
O CON~
H ~

3.86 g (18.7 mmol) of DCC were added in portions in the course of 15 min to a stirred solution, cooled to 0C, of 5.01 g (17.2 mmol) of the compound from Example I, 3.42 g (15.6 mmol) of the compound from Example IV and 2.99 g (19.5 mmol) of HOBT in 50 ml of anhydrous dichloro-methane. The reaction mixture was stirred in a thawing ice bath for 16 h, then the resulting precipitate was removed by filtration. The filtrate was washed twice with 50 ml each of saturated sodium hydrogencarbonate solution and NaCl ~olution and dried over sodium sulphate. After evaporating the solvent in vacuo and chromatography of the crude product on 500 g of silica gel (dichloro-methane : methanol s ~mmonia = 9:1:0.1), 5.78 g t75 % of theory) of the title compound were obtained as a hard L~ A 27 644 - 36 -~oam .
R~ = 0.47 (dichloromethane:methanol:ammonia - 9:1:0.1) MS (EI, 70eV) m/e = 492 (M)+

Example 3 Diethyl 1-{(4S)-4-r(ter~-butoxycarbonyl)amino]-5-cyclo hexyl-2-pentenoyl}-~2~,S)-2-tpyrrolid.inyl)-phosphonal:e ~0 BOC~ -N~p O O=P(oc2H5)2 90 g (0.66 mol) of HOBt and, hfter it had dissolved, 2S9 g (0.64 mol) of 1-cyclohexyl-3-(2-morpholinoethyl)-carbodiLmide-metho p-toluenesulphonate (CMCT) were added at 0C to a ~olution of 180 g (0.61 mol) of the compound from Example II in 900 ml of anhydrous dimethylfonmamide.
The cooling bath was removed and the mixture was then stirred at room temperatuxe for 2 h. It was then cooled again to 0C and a solution of 138 g (0.66 mol~ of diethyl 2tR,5)-2-(pyrrolidinyl)-pho~phonate in 300 ml of dimethylformamide and 161 ml (2.23 mol) of N-methyl-morpholine was added dropwise. The cooling bath was removed and the mixture was stirred at room temperature for 15 h. The reaction mixture was then concentrated in vacuo and the residue was partitioned between 1 1 of water and 1 1 o~ ethyl acetate. ~he aqueous phase was Le A 27_644 - 37 -"` 2~4~1~3 extracted with 500 ml of ethyl acetate, and the combined organic extracts were washed with 500 ml of water and dried over sodium sulphate. After evaporating the solvent in vacuo and chromatography of the crude product on 8 kg of silica gel ldichloromethane : methanol 94:6), 236 g (80 % of theory) of the title compound were obtained as a Yiscous oil.
R~ = 0.24 (dichloromethane : methanol 95:53 MS (FAB): m/e a 487 (M+H)+, 509 (M~Na)~
Diastereomer ratio: 1:1.16 (HPLC) Example 4 1-{(4S)-4-[(tert-Butoxycarbonyl)amino]-5-cyclohexyl-2-pentenoyl}-L-proline benzyl ester ~OC -As described for Example 3, 1.10 g (67 % of theory) of the title compound were obtained as an oil from 1.00 g (3.37 mmol) of the compound from Example II and 0.82 g (3.37 mmol) of L-proline benzyl ester hydrochloride after chromatography of the crude product on 94 g of silica gel (toluene s ethyl acetate - 15:85).
R~ = ~.48 ~toluene s ethyl acetate 1:9) MS (FAB)s m/e ~ 485 (M+H)~

Le A 27 644 - 38 -xample 5 1-{(4S-4-[(tert-Butoxycarbonyl~amino]-5-cyclohexyl-2-pentenoyl}-L-proline methyl ester soc-H ¦¦ ~

As described for Example 3, 866 mg (63 % of theory) of the title compound were obtained as a foam from 1.00 g (3.37 mmol) of the compound from Example II and 0.61 g (3.71 mmol) of L-proline methyl e~ter hydroohloride after chromatography of the crude product on 73 g of silica gel tdichloromethane : methanol 95:5).
Rf = 0.32 tdiehloromethane : methanol - 95:5) MS (DCI, NH3): m/e = 409 (M+H)+

Example 6 Diethyl l-[(4S)-4-amino-5-phenyl 2-pentenoyl]-(2R,S)-2~(pyrrolidinyl)phosphonate trifluoroaceta~e ~ h 2 CF3COOH x HzN ~ ~ ~
O O=P(oczH5)2 Le A 27 644 - 39 -2~31~3 2.0 ml of trifluoroacetic acid were added to a solution, cooled to 0C, of 465 mg (0.96 mmol) of the compound from Example 1 and 104 ~1 (0.96 mmol) of ani~ole in 1 ml of anhydrous dichloromethane. ~he cooling bath was removed and the mixture was then stirred at room temperature fox 20 min. 2 ml of toluene were added and the mixture was concentrated in vacuo. 2 ml of toluene were added to the residue and the mixture was again concentrated in vacuo.
This process was repeated twice more. After drying in a high vacuum, 551 mg (94 % of theory) of the title com-pound were obtained as an oil.
Rr = 0 . 31 (dichloromethane : methanol 9:1) MS (FAB): m/e = 381 (M+H)t, 403 (M+Na)+, 419 (N~R)t.

Example 7 Diethyl l-t(4S)-4-amino-5-cyclohexyl-2-pentenoyl]-(2R,S)-2-(pyrrolidinyl)phosphonate trifluoroacetate ~O
ZCF3COOH x H2N~ ~l o o=P(OC2~15 )2 As described for Example 6, 12.62 g (99 % of theory) of the title compound were obtsined as an oil from 10.10 g (20.76 mmol) of the compound from Example 2 in the pre~ence of 6.30 ml of anisole.
R~ = 0.30 (dichloromethane : methanol 9:1) Le A 27 644 - 40 -20a~ 3.

MS (FAB): m/e = 387 (M+H)+, 409 (M+Na)'.

Example 8 l-t(4S)-4-Amino-5-cyclohexyl-2-pentenoyl]-L-proline benzyl ester trifluoroacetate ~ 1~
2 CF3COOH x H2 ~

A4 described for Example 6, 947 mg (81 % of theory) of the title compound were obtained as Colo~less crystals from 927 mg (1.91 mmol) of the compound from Example 4 in the presence of 0.4 ml of ani~ole and trituration of the crude product in ether.
Melting point: 128C
F~ = 0.58 (dichloromethane : methanol 4:1 MS (DCI, NH3): m/e = 385 (M+H)~.

Example 9 1-t(4S)-4-Amino-5-cyclohexyl-2-pentanoyl]-2-proline methyl ester trifluoroacetate 2CF3cooH x H2 ~ , ~

Le A 27 644 - 41 -2~31~3 As described for Example 8, 568 mg (72 % of theory) of the title compound were obtained as colorless crystals from 601 mg (1.47 mmol) of the compound from Example 5.
Melting point: 193C
5 Rf = 0.24 ~dichloromethane : methanol 9:1) MS (FAB): m/e = 309 (M+H)+.

Example 10 1-[(4S)-4-Acetylamino-S-cyclohexy1-2-pentenoyl]-L-proline methyl ester ,~

29 ~1 (0.31 mmol) of acetic anhydride were added dropwise to a solution, cooled to 0C, of 134 mg (0.25 mmol) of the compound from Example 9 in 1 ml of anhydrous dimethylformamide and 110 ~1 (0.99 mmol) of N-methyl-lS morpholine. After 15 min at 0C, the mixture was pouredinto a mixture of 20 ml of ice-cold sodium bicarbonate solution and 10 ml of ethyl acetate and thoroughly stirred. The organic phase was separated off, the aqueous phase W88 extracted with 10 ml of ethyl acetate and the combined organic extracts were dried over magne~ium sulphate. After evaporating the solvent in vacuo and repe~tedly dissolving and concentrating again using Le A 27 644 - 42 -toluene and dichloromethane, 87 mg ~99 % of theory) of the title compound were obtaine~ as a colo~less foam.
Rf = 0.44 (dichloromethane : methanol 9:1) MS (DCI, NH3): m/e = 351 (M+H)t As described for Example 10, the following product3 were obtained (Table 1) starting from the compounds of Example 7 [R2 = P(O)(OC2H5)2], ~xample 8 (R2 = COO CH2-Ph) or Example 9 (R2 = COOCH3) by re ction with an acylating agent (W'COX) after chromatography sn ~ilica gel in the eluent mixture indicated.

Le A 27 644 - 43 -" 2~3~

a~ o ~
_ _ _ ~ ~ ~ ~ _ _ ._ ________ _ 110 O` 0 N
~" -- N N O _ _ _ OOOOOOOO O

_- 1~
-- O N u ~ ~D O

_ ~P
_ ~ ~O - O u~ o~
C~ ~=0 :~ s e ~ ~ e~~N ~N ~N

N N
N N

t~

~ _ N 1~

Le A 27 644 - 44 -5~ o ~ O

i-l N ~ 1 N --I _ N t~ N N N N
o oe~ o ~ o o o o c~ o o c~

N N ~a N N N ~Y N
~; ~N ~ N N 1~ N N N N
8 ~, u u ~ ~ ~ ~ ~ ~ _ ,C ~ =! sN --c ~ t~ q O b , c~`
C X N N N ~I N N N N N N

Le A 2? ~4 - 45 -20~31~3 Example 3_ Diethyl 1-[(4S)-5-cyclohexy1-4-(2-indolylcarbonyl)amino-2-pentenoyl (2R) and (2S)-2-(pyrrolidinyl)phosphonate ~0~
H ll o o2p(o~ 2H5~2 115 mg ~O.85 mmol) of HOBt and 342 mg (O.80 mmol) of CMCT
were added a~ 0C to a stirred solution of 125 mg (0.77 mmol) of indole-2-carboxyl1c acid in 1.5 ml of dimethylformamide. The cooling bath was removed and the mixture wa~ ~tirred at room temperature for 2 h. It was then cooled again to O~C and a solution of 430 mg (0.70 mmol) of the compound from Example 7 and 0.31 ml (2.8 mmol) of N-methylmorpholine in 1.5 ml of dimethyl-formnmide was added dropwise and the mixture was ~tirred in a thawing ice bath for 16 h. The reaction mixture was then concentrated in vacuo and the residue was parti-tioned betwsen 25 ml o~ water and 25 ml of ethyl acetate.
The aqueou~ pha~e was extracted with 20 ml of ethyl acetate and the combined organic Qxtracts were dried over magne~ium sulphate. After evaporating the solvent in vacuo and chromatography of the residue on 90 g of silica gel (dichloromethane t methanol 95s5), 118 mg (29 ~ of theory) of the non-polar diastereomer were obtained as a foam, Rr = 0.29 (diohloromethane s methanol 95s5) Le A ____~44 - 46 -204~1d~3 MS (FAB): m/e = 530 (M+H)+
and additionally 159 mg (43 ~ of theory) of the polar diastereomer as a foam.
Rf = O . 24 (dichloromethane : methanol 95:5) S MS (FAB): m/e = 530 (M~H)+.

As described for Example 30, the following products of the general formula (I) (Table 2) were obtained by reaction of the trifluoroacetates indicated with the appropriate carboxylic acids:

Le A 27 644 - 47 -2~31~

~ ~ ~ 0 u~
a~.o O~

X~-- ~q N ~ -- N

N N
~E3 . ~ O

N N N N N
~;~ 8 o ~ o o o '...~ ~
:-- ~ S N ~ ~ S

a ~s ~_ C c o Z r~

Le A 27 644 - 48 -2~3~

N $7 O 1~ 0 1`. u~ u~
h ~ ~n In ~o u~ u) v> U, u~

Ei _ _~
+~

_ _ -- o N ,~, N N
+ O O O O O O O O

N
-- ~ ~ I~ O V- t~ ~ ~O .
~ r~l r~ I
~ E~

a) dP
~_ Il- ~ N U~ t~ ~ 1~ r~

N N ~ N N
N N N 3 3 ~ N N
O O ~ ro -- -- -- O O
-- -- ~L Z j~ 2 ; a o "
O ~ O ~ ~ ~ O O

p: i~ p p ~
~ t~l N N N N N N N

~ al O '¢ D n~ L V~
1 ,~, O o ~Ia ~ Z :1:
~J ~ r~ V` 1:~ --' N t~
1~ .
C X
C~ ~

27 64~ - ~19 -~ ~ ~ ~ ~ ~ ~ ~

h o co u~
P:-- ~ ~ o o oo o o o ~3 ~ ~ ~ C~
~ ~ ~_ _I
. _ ~q rq ~n ~ u~
t~3 N N N t~3 U~
N ~ N ~ N

~ o o o o o P~ Q Q Q Q
,. ~ C~
P:

a a ~r l N ~1~ N N N
C ~ _ _ ~_ _ ~ ~i3 Le A 2? 644 - 50 -20~3~

C ~

~.~ .. ~ ~2 ~:-- o~ ow a~ o _ ~_ ~.,, ~ ~ _ _ o ~

P~ ~ o o h 3 ~~~ U~ U~ j ~o X~ ~ ~ 0~

.,./ _ N ~
p., ~r ~ ~ ~

N N a 8 P: _ ~. _ O O

N ~ ~? -- ; ~

c ~ . . ~ ~9 C: 5~ ~ ~ ~ W ~
Z ~N ~N 8 a~3 ~ x a ~
Le A 27 644 - 51 -2~31~3 ~ N ~

a~ .O O~

~1 o~ ~ ~'I N N N

+
_ _ ~~r N o ~" ~ o GdP 5 ", ~ .

~ t~~ N t~ N e~ <~

--~ UUN UN N SN -- N

æ æ 5 ~? ~? U U

` e 5 ~q S ~

Nt'l ~ "~

Le A 2? 644 -52 -2 ~ 3 Example S9 Lithiummonoethyll-{(4S)-4-t(tert-butoxycarbonyl)amino]-5-phQnyl-2-pentenoyl}-(2Rrs)-2-(pyrrolid$nyl)pho~phonate f'P" 1--BOC_~
' I - OC2H5 OLi 74 mg ~0.36 mmol) of anhydrous lithium $odide and 30 pl (0.36 mmol) of pyridine were added to a stirred solution of 78 mg (0.16 mmol) of the compound from Example 1 in 2 ml of anhydrous acetonitrile and the mixture was heated under reflux for 2.5 h. It was then allowed to cool, S ml of toluene were added and the mixture was concentr~ted to dryness in V8CUO. This proces~ was repeated twice before the crude product was filtered on 1.2 g of silica gel (dichloromethane : methanol 4:1). The product-containing fractions were concentrated, dissolved in 1 ml of water and ad~usted to pH 7.5 by ~ddition of aqueous lN LiOH
solution. Chromatography of thi~ aqueous solution on a Merck Lobar LiChroprep RP-8 re~dy-to-u~e column, size A
(water with ~n increasing content of acetonitrile) and freeze-drying of the product fractions gave 24 mg (32 4 of theory) of the title compound a~ a cOlO~less lyophil-isate.
F~ - 0.24 (d~chloromethane s m~thanol 4sl) ~S (FAB)s m/Q ~ 459 (N+H)~, 465 (M+Li)~.

Le A ~7 Ç4~ _ 53 _ . t,~ 3 Example_60 Sodium mono~thyl 1-[(4S)-4-amino-5-phenyl-2-pentenoyl~-(2R,S)-2-tpyrxolidinyl~pho~phonat~
Fh ~2N~
O O~P-oc2Hs ON~
0.60 ml (4.~ ~mol - 2.2 aquival0nts) of trimethyliodo-silane wa~ 810wly ~dded dropwise to a stirred solution, cooled to 0Ct of 961 mg (2.0 mmol~ of the compound from E~ample 1 in 4 ml of anhydrous dichloromethane. The mixture wa~ stirred at O~C for 1.5 h and gently concen-trated in vacuo. The residue was partitioned b~tween 20 ml of ethyl acetate and 20 ml of water. The cold aqueou~ phase wa~ ad~usted to pH B with lN NaOH. The organic phase wa ~ep~r~ted off and disc~rded. The aqueous phase wa~ concentrated in vacuo to a vol~me of 5 ml and chromatographed on a ~erck Lobar LiChroprep RP-8 ready-to-use colu~n, size B (water with an increa~ing content of ~cetonitrile). After free~e-drying the prsduct fraction3, 355 mg (51 % of theory) of the ~itle compound were obtained a8 a lyophilizate.
R~ = 0.18 (dichloromethane s ~ethanol 4:1) MS (FAB): m/e ~ 375 (M+H)~.

e ~ 27 ~44 54 -~9D~2~
Diethyl 1-{(4S)-4-[(ter~-butylaminocarbonyl)amino]-5-cyclohexyl-~-pentenoyl}-(~R,S)-2-(pyrrolidinyl~-phos-phonate (H3c)3c-N-cO-o o~PtOC2H5)z 70 ~1 (0.59 mmol) of ~ert-butyl isocyanate were added dropwise to a ~tirred solution of 330 mg (O.54 mmol~ of the compound from Example 7 and 150 ~1 (1.07 mmol) of triethylamine in 3 ml of anhydrous dichloromethane. After 30 min at room temperature, 3 ml of toluene were added and the reaction mixture was cow entrated in vacuo. After chromatography of th~ re~idue on 41 g of silica gel (dichloromethane : methanol 93:7), 144 mg (55 % of theory~ of the ti~le compound were obtained ~ a colorless les~ hard foam.
Rr ~ 0.36~ 0.34 (dichlorome~hane s meth~nol 93:7) MS (FAB): m/e = 486 (M+H)t, 508 (M+Na)+

As de~cribed ior Fx~mple 61, the following product~
(Table 4) were obtained by reacting the compound from ~xample 7 w.ith varLous isocy~nate~:

e A 27 644 - 55 ¢~
r1 N

~_ _ - N
_~ ~ O C~ O

_ _ ~ ~ 5 ~ r~ , ~_ .
. ~, u~

--~= ~ N UN ~ CN N N aJ
a~ o ~ u (:
Q Q Q = ~

X[~ O

Le A 27644 - 56 -2 ~ 3 It will be understood that the specification and examples are illustrative but not limitative of the present invention and that other embodiments within the spirit and scope of the invention will suggest themselves to those skilled in the art.

Le A 27 644

Claims (15)

1. A pyrrolidine- or piperidine-substituted pseudo-peptide of the formula (I) in which W - is hydrogen or an amino protecting group, or - is straight-chain or branched alkyl or alkenyl in each case having up to 6 carbon atoms, which are optionally substituted by aryl having 6 to 10 carbon atoms, or - is a group of the formula R3-Co-, R5R4N-CO-or R6-SO2-, in which R3 - denotes hydrogen, trifluoromethyl or straight-chain or branched alkoxy having up to 8 carbon atoms or alkyl having up to 18 carbon atoms, which are optionally substituted by aryl having 6 to 10 carbon atoms or pyridyl, or - denotes aryl having 6 to 10 carbon atoms, which is optionally substituted by halogen, tri-fluoromethyl, trifluoromethoxy or by straight-chain or branched alkyl having up to 8 carbon atoms, - denotes cycloalkyl having 3 to 7 carbon atoms, Le A 27 644 -58-or - denotes indolyl, pyridyl, morpholino or piper-azinyl, or - denotes a radical of the formula in which R7 - denotes phenyl or naphthyl;
R8, R9 and R10 are identical or different and denote straight-chain or branched alkyl having up to 17 carbon atoms, which is optionally substituted by phenyl or naphthyl r or denote aryl having 6 to 10 carbon atoms, which is optionally substituted by alkyl having up to 4 carbon atoms, m - denotes n number O, 1 or 2, T - denotes morpholino or cyclohexyl, p - denotes a number 1, 2 or 3, Y and Y' are identical or different and denote the CO or SO2 group, Le A 27 644 -59-t - denotes a number 0 or 1, R11 and R11' are identical or different and denote hydroxyl or alkoxy having up to 8 carbon atoms, s - denotes a number 1 or 2, R4 and R5 are identical or different and - denote hydrogen or - denote aryl having 6 to 10 carbon atoms, which is optionally substituted by straight-chain or branched alkyl having up to 6 carbon atoms or halogen, or - denote cycloalkyl having 3 to 7 carbon atoms, or - denote straight-chain or branched alkyl having up to 18 carbon atoms, which is optionally substituted by pyridyl, R6 - denotes straight-chain or branched alkyl having up to 8 carbon atoms, A, B and D are identical or different and each - is a direct bond or - is a radical of the formula , or , in which Le A 27 644 -60-x and x' are identical or different and denote the number 1 or 2 and r - denotes the number 0 or 1, or - is a group of the formula in which z - denotes the number 0 or 1, R12 denotes hydrogen or straight-chain or branched alkyl having up to 8 carbon atoms, R13 denotes cycloalkyl having 3 to 8 carbon atoms or aryl having 6 to 10 carbon atoms or hydro-gen, or - denotes straight-chain or branched alkyl having up to 8 carbon atoms, which is optionally substituted by alkylthio having up to 6 carbon atoms, hydroxyl, mercapto, guanidyl or by a group of the formula -NR14R15 or R16-OC-, in which R14 and R15 are identical or different and denote hydrogen, straight-chain or branched alkyl having up to 8 carbon atoms or phenyl, and R16 denotes hydroxyl, benzyloxy, alkoxy having up to 6 carbon atoms or the abovementioned Le A 27 644 group -NR14R15, or which is optionally substituted by cyclo-alkyl having 3 to 8 carbon atoms or by aryl having 6 to 10 carbon atoms, which is optionally substituted by hydroxyl, halogen, nitro, alkoxy having up to 8 carbon atoms or by the group -NR14R15, or which is optionally substituted by a 5- or 6-membered nitrogen-containing heterocycle or indolyl, in which the corresponding -NH func-tions are optionally protected by alkyl having up to 6 carbon atoms or by an amino protecting group, R1 - is cycloalkyl having 3 to 8 carbon atoms, - is straight-chain or branched alkyl or alkenyl having up to 10 carbon atoms, which are optionally substituted by cycloalkyl having 3 to 8 carbon atoms or aryl having 6 to 10 carbon atoms, which is optionally substituted by halogen, nitro, hydroxyl, amino or straight-chain or branched alkoxy having up to 6 carbon atoms, n - is the number 1 or 2, R2 - is a radical of the formula O=?R11R11', -CO-R17 or -CO-NR4R5, and Le A 27 644 -62-R17 - is straight-chain or branched alkoxy having up to 8 carbon atoms, which is option-ally substituted by phenyl or a physiologically acceptable salt thereof.

2- A pseudopeptide or salt thereof according to claim 1, in which W - represents hydrogen, tert-butyloxycarbonyl (BOC), 9-fluorenylmethyloxycarbonyl (Fmoc) or benzyloxycarbonyl (Z), or - is straight-chain or branched alkyl or alkenyl in each case having up to 4 carbon atoms, which are optionally substituted by phenyl, or - is a group of the formula R3-CO- or R5R4N-CO-, in which R3- denotes hydrogen, trifluoromethyl or straight-chain or branched alkoxy having up to 4 carbon atoms or alkyl having up to 16 carbon atoms, which are optionally substituted by phenyl, naphthyl or pyr-idyl, or - denotes phenyl or naphthyl, each of which is optionally substituted by fluorine, chlorine, trifluoromethyl, trifluorometh-oxy or by straight-chain or branched alkyl having up to 6 carbon atoms, - denotes cyclopropyl, cyclopentyl, Le A 27 644 cyclohexyl, indolyl, pyridyl, morpholino or piperazinyl, or - denotes a radical of the formula or in which Y - denotes the CO or SO2 group, R7 - denotes phenyl or naphthyl, R8, R9 and R10 are identical or different and denote straight-chain or branched alkyl having up to 15 carbon atoms, tolyl, phenyl or naphthyl, m - denotes a number 1 or 2, R4 and R5 are identical or different and - denote hydrogen, - denote phenyl or naphthyl, each of which is optionally substituted by straight-chain or branched alkyl having up to 4 carbon atoms, fluorine or chlorine, - denote cyclopropyl, cyclopentyl or cyclo-hexyl, or denote straight-chain or branched alkyl having up to 16 carbon atoms, which is optionally substituted by pyridyl, A, B and D are identical or different and - are a direct bond, Le A 27 644 -64-- are : proline, - are a radical of the formula or , in which r denotes the number 0 or 1, and x' denotes the number 1 or 2, - are a group of the formula in which z - denotes the number 0 or 1, R12- denotes hydrogen or straight-chain or branched alkyl having up to 6 carbon atoms, R13- denotes cyclopentyl, cyclohexyl, phenyl or hydrogen, - or denotes straight-chain or branched alkyl having up to 6 carbon atoms, which is optionally substituted by hydroxyl, carboxyl or H2N-CO-, or is subtituted by cyclohexyl, naphthyl or phenyl, each of is optionally substituted by fluorine, hydroxyl, nitro Le A 27 644 -65-or alkoxy having up to 4 carbon atoms, or is substituted by indolyl, imidazolyl, pyridyl, triazolyl or pyrazolyl, where the corresponding -NH functions are optionally protected by alkyl having up to 4 carbon atoms or by an amino protecting group, R1 - is cyclopropyl, cyclopentyl or cyclo-hexyl, or - is straight-chain or branched alkyl or alkenyl having up to 8 carbon atoms, which are optionally substituted by cyclopropyl, cyclo-butyl, cyclopentyl, cyclohexyl, cycloheptyl or phenyl, each of which is optionally substituted by fluorine, chlorine, bromine, nitro, hydroxyl or amino, n - is the number 1 or 2, R2- is a radical of the formula O-?-R11R11' -CO-R17 or -CO-NR4R5 , in which R11 and R11' are identical or different and denote hydroxyl or straight-chain or branched alkoxy having up to 6 carbon atoms, and R17 - denotes straight-chain or branched alkoxy in each case having up to 6 carbon atoms, which is optionally substituted by phenyl.

Le A 27 644

3. A pseudopeptide or salt thereof according to claim 1, in which W - is hydrogen, tert-butyloxycarbonyl (BOC) or benzyloxycarbonyl (Z), or - is allyl or benzyl, - is a group of the formula R3-CO- or R5R4N-CO-, in which R3 - denotes hydrogen, trifluoromethyl or straight-chain or branched alkyl having up to 14 carbon atoms which are optionally substituted by phenyl, naphthyl or pyri-dyl, - denotes phenyl or naphthyl, each of which is optionally substituted by fluorine, chlorine, trifluoromethyl, trifluorometh-oxy or straight-chain or branched alkyl having up to 4 carbon atoms, - denotes cyclopropyl, cyclopentyl, cyclo-hexyl, indolyl, pyridyl, morpholino or piperazinyl, or - denotes a radical of the formula or Le A 27 644 -67-in which Y - denotes the CO or SO2 group, R7- denotes phenyl or naphthyl, R8,R9 and R10 are identical or different and denote straight-chain or branched alkyl having up to 14 carbon atoms, tolyl, phenyl or naphthyl, m - denotes a number 1 or 2, R4 and R5 are identical or different and - denote hydrogen, - denote phenyl or naphthyl, each of which is optionally substituted by methyl, fluorine or chlorine, - denote cyclopropyl, cyclopentyl or cyclo-hexyl, or - denote straight-chain or branched alkyl having up to 14 carbon atoms, which is optionally substituted by pyridyl, A, B and D are identical or different and each - is a a direct bond, - is proline, or - is a group of the formula in which z - denotes the number 0 or 1, R12 - denotes hydrogen or methyl, R13 - denotes cyclopentyl or cyclohexyl, - or denotes straight-chain or branched Le A 27 644 -68-alkyl having up to 4 carbon atoms, which is optionally substituted by hydroxyl, carboxyl or H2N-CO-, or is substituted by cyclohexyl, naphthyl or phenyl, each of which is optionally substituted by fluorine, chlorine or alkoxy having up to 4 carbon atoms, or is substituted by amidazolyl, tri-azolyl, pyridyl or pyrazolyl, where the NH function is optionally protected by methyl, benzyloxymethylene or t-butyl-oxycarbonyl (Boc), R1 - is cyclopentyl or cyclohexyl, or - is straight-chain or branched alkyl having up to 6 carbon atoms, which is option-ally substituted by cyclopropyl, cyclopentyl, cyclohexyl or phenyl, each of which is optional-ly substituted by hydroxyl, n - is the number 1 or 2, R2 - is a radical of the formula O=?R11R11' -CO-R17 or -CO-NR4R5 , in which R11 and R11' are identical or different and denote hydroxyl or straight-chain or branched alkoxy having up to 4 carbon Le A 27 644 -69-atoms, and R17 - denotes straight-chain or branched alkoxy in each case having up to 4 carbon atoms, which is optionally substituted by phenyl.

4. A pseudopeptide or salt thereof according to claim 1, wherein W is hydrogen, tert- butoxycarbonyl, acetyl, trifluoro-acetyl, butyryl, 2,2-dimethylpropionyl, 4-methylbutyryl, tetra-decanoyl, benzoyl or 4-methylbenzoyl, A, B and D are each bonds, R1 is benzyl or cyclohexylmethyl and R2 is diethylphosphonate, 2-(pyrid-2-yl)ethylaminocarbonyl, benzyloxycarbonyl or methoxy-carbonyl.

5. A pseudopeptide or salt thereof according to claim 1, wherein W-A-B-D- together form a 2-indolylcarbonyl, BOC-Asn, BOC-Val, BOC-Ile, BOC-Phe-Asn, BOC-Phe-Ile, BOC-Phe-His, BOC-Ser-Phe-Asn, CH3(CH2)12 CO-Phe-Asn, CH3(CH2)12CO-Val-Asn, BOC-Phe-Val, CH3(CH2)12CO-Val-Phe-Val, CH3(CH2)12CO-Phe or CH3(CH2)12CO-Val group or a group of formula or R1 is benzyl or cyclohexylmethyl and R2 is benzyloxycarbonyl, methoxycarbonyl, diethylphosphonate, sodium monomethyl phosphonate or 2-(pyrid-2-yl) ethylaminocarbonyl.

6. A pseudopeptide or salt thereof according to claim 1 wherein W-A-B-D- together form hydrogen, H-Asn, H-Ile, H-Phe-Asn, H-Ser-Phe-Asn, BOC or tert-butylaminocarbonyl, R1 is benzyl or cyclohexylmethyl and R2 is diethylphosphonate, sodium monomethyl-phosphonate, lithium monomethylphosphonate or 2-(pyrid-2-yl) ethylaminocarbonyl.

7. A pseudopeptide or salt thereof according to claim 1 wherein W is cyclohexylaminocarbonyl, dodecylaminocarbonyl, 4-chlorophenylaminocarbonyl, naphth-1-ylaminocarbonyl, aminocarbonyl, R1 is cyclohexylmethyl and R2 is diethylphosphonate.

8. The compound sodium monomethyl 1-[(45)-4-amino-5-phenyl-2-pentenoyl]-(2R,S)-2-(pyrrolidinyl) phosphonate.

9. A pseudopeptide or a salt thereof according to any one of claims 1 to 7 wherein n is 1.

10. A process for preparing a pseudopeptide or salt there-of according to any one of claims l to 7 which process comprises (a) condensing a compound of formula (Ia) (Ia) in which n, R1 and R2 are as defined in any one of claims 1 to 7 with a compound of formula (II) W-A-B-D-OH (II) in which W ,A, B and D are as defined in any one of claims 1 to 7 and the carbonyl group is activated, if necessary; or (b) reacting a compound of formula (Ia) as defined above with a compound of formula (III) or (IV) W-X (III) (W")20 (IV) in which W is as defined in any one of claims 1 to 7, X represents a halogen and W" represents the group CF3CO or CH3CO; or (c) condensing a compound of formula (V) or (VI) (V) (VI) in which R1, W, A, B and D are as defined in any one of claims 1 to 7, W' represents an amino protecting group, and the carboxylic acid group is activated, if necessary, with a compound of formula (VII) (VII) in which n and R2 are as defined in any one of claims 1 to 7 followed, if necessary by removal of the protecting group W'; and, if required, converting an obtained compound of formula I into a physiologically acceptable salt thereof.

11. A retroviral composition comprising a retrovirally effective amount of a pyrrolidine- or piperidine- substituted pseudopeptide according to any one of claims 1 to 8, or a physiologically acceptable salt thereof, and a pharmaceutically acceptable diluent.

12. The use for combating retrovira of a pyrrolidine- or piperidine-substituted pseudopeptide according to any one of claims 1 to 8, or a physiologically acceptable salt thereof.

13. A commercial package containing as active pharmaceuti-cal ingredient a pyrrolidine- or piperidine-substituted pseudo-peptide according to any one of claims 1 to 8 or a physiologically acceptable salt thereof, together with instructions for the use thereof for combating retrovira.

14. A compound of formula (Ia) (Ia) wherein R1, R2 and n are as defined in claim 1, or a salt thereof.

15. A process for preparing a compound of formula Ia as defined in claim 14, which process comprises reacting a compound of formula (V) (V) in which R1 is as defined in claim 14, W' is an amino protecting group and the carboxyl group is activated, if necessary, with a compound of formula (VII) in which R2 and n are as defined in claim 14.