Peptides derived from trifluoromethylketones

Compounds of formula (I): ##STR1## wherein: R.sub.1 represents an unsubstituted or substituted alkyl, PA1 R.sub.2 represents an unsubstituted or substituted alkyl, PA1 A represents, with the nitrogen and carbon atoms to which it is attached, a nitrogenous mono- or polycyclic structure, and PA1 B represent any one of the radicals a through j as defined in the description, and medicaments containing the same, are useful as human leucocyte elastase inhibitors.

The present invention relates to new peptides derived from trifluoromethyl 
ketones. 
These new peptide compounds have inhibitory properties for human leucocyte 
elastase. 
Elastin is an elastic fibrous protein of the connective tissue of 
vertebrates. It is present in vascular walls, skin, lungs, cartilages, 
ligaments and other tissues. Elastases are enzymes capable of solubilizing 
fibrous elastin. Human leucocyte elastase is a serine protease, which is 
found in the active form in azurophile granules of the polymorphonuclear 
neutrophile. It is a glycoprotein of 25 to 30 kDa formed from 218 amino 
acids. Human leucocyte elastase (HLE) solubilizes fibrous elastin but also 
cleaves other proteins of the extracellular matrix (collagens, 
fibronectin, proteoglycans, and the like) and hydrolyzes and inactivates a 
certain number of plasma proteins (clotting factor, immunoglobulin, 
complement, and the like). The elastolytic activity is controlled and 
regulated by natural inhibitors (.alpha.-1-antitrypsin, 
.alpha.-2-macroglobulin or bronchial inhibitor). 
Reversible or irreversible inhibitors of human leucocyte elastase have been 
described in the literature for the treatment of physiopathologic 
situations where its role has been mentioned (D. A. Trainor, TIPS, 8, 
303-307, 1987). 
These pathological states can be pulmonary emphysema, rheumatoid arthritis, 
degenerative diseases of the connective tissue such as atherosclerosis (J. 
G. Bieth, "Elastases: Catalytic and Biological Properties" in "Regulation 
of matrix accumulation"--R. P. Mecham--Academic Press, N.Y., 217-320, 
1986), acute respiratory distress syndrome in adults (P. M. Suter et al., 
Am. Rev. Respir. Dis., 145, 1016-1022, 1992), cystic fibrosis (K. C. Meyer 
et al., Am. Rev. Respir. Dis., 144, 580-585, 1991), chronic bronchitis (J. 
A. Nadel, Respiration, 58 (suppl. 1, 3-5), 1991), glomerulonephrites (E. 
Sanders et al., Renal. physiol., 3, 355-359, 1980), psoriasis (J. 
Schalkwijk et al., Br. J. Dermatology, 122, 631-644, 1990), tissue lesions 
arising during ischemia/reperfusion procedures (F. A. Nicolini et al., Am. 
Heart J., 122-1245, 1991 and C. R. B. Welbourn et al., Am. J. Physiol., 
260, 1852-1856, 1991). It may also play a role in phenomena of normal or 
pathological/tumor invasion cell migration (J. G. Bieth cited above). 
Recently, peptides derived from trifluoromethyl ketones have been described 
as HLE inhibitors. This is more particularly the case for the compounds 
described in Patents EP 189,305 and EP 369,391. 
The present invention more specifically relates to the compounds of formula 
(I): 
##STR2## 
in which: 
R.sub.1 represents a linear or branched (C.sub.1 -C.sub.6)alkyl radical 
which is unsubstituted or substituted by a (C.sub.3 -C.sub.7)-cycloalkyl, 
phenyl, amino or benzyloxycarbonylamino, 
(3,5-di-tert-butyl-4-hydroxy)benzylcarbonylamino, 
(3,5-di-tert-butyl-4-hydroxy)phenylthio or 
3-[(3,5-di-tert-butyl-4-hydroxy)phenylthio]propylcarbonylamino radical, 
R.sub.2 represents a linear or branched (C.sub.1 -C.sub.6)alkyl radical 
which is unsubstituted or substituted by a (C.sub.3 -C.sub.7)-cycloalkyl 
or phenyl radical, 
A represents, with the nitrogen and carbon atoms to which it is attached, a 
2-azabicyclo[2.2.2]octane, 2-azabicyclo[2.2.1]heptane, perhydroindole, 
perhydroisoindole, indoline, isoindoline, perhydroquinoline, 
perhydroisoquinoline, 1,2,3,4-tetra-hydroquinoline, 
cyclopenta[b]pyrrolidine or 1,3-thiazolidine ring, 
represents any one of the following radicals: 
##STR3## 
which: R.sub.3 represents a hydrogen atom or a linear or branched (C.sub.1 
-C.sub.6)alkyl radical which is unsubstituted or substituted by a phenyl 
radical, 
X, Y, which are different, represent CO or SO.sub.2, 
Z represents 
a linear or branched (C.sub.1 -C.sub.6)alkyl radical which is substituted 
by one or two, identical or different, (C.sub.3 -C.sub.7)cycloalkyl or 
(C.sub.1 -C.sub.4)trihaloalkyl radicals, 
an adamant-1-yl radical, 
a phenyl radical which is unsubstituted or-substituted by one or a number 
of, identical or different, halogen atoms or linear or branched (C.sub.1 
-C.sub.6)alkyl, hydroxyl, linear or branched (C.sub.1 -C.sub.6)alkoxy, 
(C.sub.1 -C.sub.4)trihaloalkyl, cyano, 1,4-dihydropyrid-4-yl 
(unsubstituted or substituted by one or a number of, identical or 
different, linear or branched (C.sub.1 -C.sub.6)alkyl or linear or 
branched (C.sub.1 -C.sub.6)-alkoxycarbonyl radicals), 
(3,5-di-tert-butyl-4-hydroxy)benzylcarbonylaminosulfonyl, 
(3,5-di-tert-butyl-4-hydroxy)benzoyloxy, (3-ethoxy-2-hydroxy)propoxy or 
[3,5-di-tert-butyl-4-ethoxymethoxy)]benzyloxy radicals, 
or, a (C.sub.1 -C.sub.4)alkenyl radical substituted by a 
[3,5-di-tert-butyl-4-(ethoxymethoxy)]phenyl or 
(3,5-di-tert-butyl-4-hydroxy)phenyl radical, 
##STR4## 
in which: R.sub.4, R.sub.5, which are identical or different, represent: a 
hydrogen atom or a linear or branched (C.sub.1 -C.sub.6)alkyl 
(unsubstituted or substituted by a phenyl group), phenyl (unsubstituted or 
substituted by one or a number of halogen atoms or linear or branched 
(C.sub.1 -C.sub.6)alkyl, linear or branched (C.sub.1 -C.sub.6)alkoxy or 
(C.sub.1 -C.sub.4)trihaloalkyl radical), 3-azabicyclo[3.3.01oct-3-yl, 
4-(2,3,4-tri-methoxybenzyl)piperazino, morpholino, pyrrolidino, 
piperidino, azetidino or 1-oxa-3,8-diaza-2-oxospiro[4.5]dec-8-yl 
(unsubstituted or substituted by a linear or branched (C.sub.1 
-C.sub.6)alkyl or linear or branched (C.sub.1 -C.sub.6)phenylalkyl 
radical) radical, or 
form, with the nitrogen atom to which they are attached, any one of the 
following rings: 
3-azabicyclo [3.3.0]octane, 
4-(2,3,4-trimethoxybenzyl)piperazine, 
morpholine, 
pyrrolidine, 
azetidine, 
3,8-diaza-1-oxa-2-oxospiro[4.5]decane which is unsubstituted or substituted 
by a linear or branched (C.sub.1 -C.sub.6)alkyl or linear or branched 
(C.sub.1 -C.sub.6)phenylalkyl radical, 
##STR5## 
in which: R.sub.3 is as defined above and 
R.sub.6 represents a hydrogen or halogen atom or a linear or branched 
(C.sub.1 -C.sub.6)alkyl, linear or branched (C.sub.1 -C.sub.6)alkoxy, 
(C.sub.1 -C.sub.4)trihaloalkyl or cyano radical, 
##STR6## 
in which: R.sub.3 is as defined above, 
T represents any one of the following groups: 
--(CH.sub.2)m--, 
--SO.sub.2 --NH--CO--, 
--CO--NH--SO.sub.2 --, 
##STR7## 
such that m is an integer between 1 and 4, 
##STR8## 
in which: X is as defined above, 
n is equal to 0, 1, 2 or 3, 
n' is equal to 0 or 1, 
R.sub.7 represents a: 
3,5-di-tert-butyl-4-hydroxyphenyl, 
3,5-di-tert-butyl-4-hydroxyphenoxy, 
or 3,5-di-tert-butyl-4-hydroxyphenylthio radical 
##STR9## 
in which p is equal to 1 or 2, 
##STR10## 
in which R.sub.3 is as defined above, 
##STR11## 
in which A is as defined above, which compounds of formula (I) comprise 
the corresponding hydrates of the COCF.sub.3 ketone functional group, 
their enantiomers, diastereoisomers and epimers and their addition salts 
with a pharmaceutically acceptable acid or base. 
Among the pharmaceutically acceptable acids, there may be mentioned, as 
non-limiting, hydrochloric, sulfuric, tartaric, maleic, fumaric, 
methanesulfonic or camphoric acids, and the like. 
Among the pharmaceutically acceptable bases, there may be mentioned, as 
non-limiting, sodium hydroxide, potassium hydroxide, tert-butylamine, 
diethylamine, ethylenediamine, and the like. 
The invention also applies to the process for the preparation of the 
compounds of formula (I), characterized in that there is used, as starting 
material, an alcohol of formula (II), the isomers of which have optionally 
been separated by a conventional separation technique: 
##STR12## 
in which R.sub.2 has the same meaning as in formula (I), which is reacted: 
a either with a protected amino acid of formula (III), the isomers of 
which have optionally been separated according to a conventional 
separation technique, by a conventional peptide coupling technique such as 
that described by W. Konig and R. Geiger (Ber., 103, 788, 1970): 
##STR13## 
in which A has the same meaning as in formula (I) and Boc represents a 
tert-butoxycarbonyl group, to lead to the compound of formula (IV): 
##STR14## 
in which A, R.sub.2 and Boc have the same meaning as above, which 
compound of formula (IV) is: 
* either deprotected by acid hydrolysis to lead to the compound of formula 
(V): 
##STR15## 
in which A and R.sub.2 have the same meaning as above, which is reacted 
with a protected amino acid of formula (VI), the isomers of which have 
optionally been separated according to a conventional separation 
technique, 
in the presence of a conventional coupling agent for peptide synthesis: 
##STR16## 
in which Boc represents a butoxycarbonyl radical and R'.sub.1 represents 
a linear or branched (C.sub.1 -C.sub.6)alkyl radical which is 
unsubstituted or substituted by a (C.sub.3 C.sub.7)cycloalkyl, phenyl or 
benzyloxycarbonylamino radical, 
to lead to the compound of formula (VII): 
##STR17## 
in which Boc, R'.sub.1, A and R.sub.2 have the same meaning as above, 
which is oxidized to lead to the compound of formula (VIII), the isomers 
of which are optionally separated according to a conventional separation 
technique, 
##STR18## 
in which Boc, R'.sub.1, A and R.sub.2 have the same meaning as above, * 
or is oxidized to lead to the compound of formula (X): 
##STR19## 
in which Boc, A, and R.sub.2 have the same meaning as above, which is 
deprotected in acid medium to lead to the compound of formula (XI): 
##STR20## 
in which A and R.sub.2 have the same meaning as above, which is reacted 
with a protected amino acid of formula (VI) as defined above, 
to lead to the compound of formula (VIII) defined above, 
b or with a protected dipeptide of formula (XII), obtained by conventional 
coupling of two amino acids in the racemic form or in the form of pure 
enantiomers, 
##STR21## 
in which Boc, R'.sub.1 and A have the same meaning as above, to lead to 
the compound of formula (VII) defined above, which is oxidized and leads 
to the compound of formula (VIII) defined above, 
which compound of formula (VIII) is deprotected in acid medium, 
to lead to the compound of formula (IX), the isomers of which are 
optionally separated according to a conventional separation technique, 
##STR22## 
in which R'.sub.1, A and R.sub.2 have the same meaning as above, which is 
reacted with an acid of formula (XIII), according to a conventional 
peptide coupling technique, 
##STR23## 
in which B has the same meaning as in formula (I), to lead to the 
compound of formula (I/a), a specific case of the compounds of formula 
(I), 
##STR24## 
in which B, A, R'.sub.1 and R.sub.2 have the same meaning as above, 
which, when R'.sub.1 represents an alkyl radical substituted by a 
benzyloxycarbonylamino group, is deprotected, if desired, by catalytic 
hydrogenation, 
to lead to the compound of formula (I/b), a specific case of the compounds 
of formula (I), 
##STR25## 
in which B, A and R.sub.2 have the same meaning as in formula (I) and 
R".sub.1 represents an alkyl substituted by an amino radical, 
which compounds of formula (I/a) and (I/b) are purified according to a 
conventional purification technique, the isomers of which are separated, 
if desired, according to a conventional separation technique, and then, if 
necessary, are converted to an addition salt with a pharmaceutically 
acceptable acid or base. 
The compounds of formula (I) have very advantageous pharmacological 
properties, in particular properties inhibitory for human leucocyte 
elastase. As such, they can be used with advantage in a certain number of 
therapeutic indications such as pulmonary emphysema, chronic bronchitis, 
acute respiratory distress syndrome in adults, cystic fibrosis, rheumatoid 
arthritis, glomerulonephritis, inflammations, ischemia reperfusion 
syndromes, phenomena of invasion and diffusion of malignant cells, 
degenerative diseases of connective tissue or skin aging. 
The inhibitory activity for human leucocyte elastase was demonstrated by in 
vitro and in vivo tests. The compounds showed greater inhibitory 
activities than the reference materials, such as chloromethyl ketone or 
dichloroisocoumarin. 
The substituents of the compounds of formula (I) have made it possible to 
add, to the inhibitory activity for human leucocyte elastase, properties 
of antiinflammatory, antiradical and/or mucoregulatory type. 
Another subject of the present invention is the pharmaceutical compositions 
containing, as active principle, at least one compound of general formula 
(I) or one of its addition salts with a pharmacologically acceptable acid 
or base, alone or in combination with one or a number of nontoxic, inert 
excipients or vehicles. 
Among the pharmaceutical compositions according to the invention, there can 
more particularly be mentioned those which are suitable for oral, 
parenteral or nasal administration, simple or sugar-coated tablets, 
sublingual tablets, gelatin capsules, troches, suppositories, creams, 
ointments, dermal gels, aerosols, drinkable and injectable phials, and the 
like. 
The useful dose varies depending on the age and weight of the patient, the 
nature and severity of the ailment and on the administration route. 
The latter can be oral, nasal, rectal or parenteral. Generally, the unit 
dose ranges between 10 .mu.g and 300 mg for a treatment taken 1 or 3 times 
per 24 hours. 
A preferential administration route for the derivatives of the invention is 
the aerosol route, in the form of a powder or of a liquid aerosol. 
The following examples illustrate the invention and do not limit it in any 
way. 
The starting materials used are known products or products prepared 
according to known procedures. The abbreviations used in the examples are 
the following: 
Abo in place of 2-azabicyclo[2.2.2]octane-3-carbonyl 
Boc in place of tert-butoxycarbonyl 
Val in place of valyl 
Phi in place of perhydroindole-2-carbonyl 
Abh in place of 2-azabicyclo[2.2.1]heptane-3-carbonyl 
Lys in place of lysyl 
Cys in place of cysteinyl 
Preparations A to V describe starting materials which are useful in the 
synthesis of the compounds of formula (I). 
Preparation A: 4-(4-Chlorobenzoylaminosulfonyl)benzoic acid 
60 mmol of 4-carbethoxybenzenesulfonamide and a solution containing 60 mmol 
of 4-dimethylaminopyridine in 40 ml of dimethylformamide (DMF) are added 
successively to a solution containing 60 mmol of parachlorobenzoic acid in 
100 ml of DMF. The whole is maintained at room temperature with stirring 
for 20 hours, filtered and then evaporated. The oil obtained is purified 
by chromatography on a silica column, using a dichloromethane/ethanol 
(93/7) mixture as solvent, and leads to ethyl 
4-(4-chlorobenzoylaminosulfonyl)benzoate which is saponified by stirring 
for 20 hours in a mixture containing 48 ml of 1N sodium hydroxide solution 
and 60 ml of ethanol. After evaporation, addition of 100 ml of water and 
then 50 ml of 1N hydrochloric acid, the expected product precipitates and 
is filtered and washed with water and dichloromethane. 
Melting point: 265.degree. C. 
Preparation B: 4-[(2-Oxotetrahydrothiophen-3-yl)aminosulfonyl]benzoic acid 
50 mmol of homocysteinethiolactone hydrochloride and 100 mmol of 
triethylamine in 30 ml of dioxane are added simultaneously at room 
temperature to 50 mmol of 4-chlorosulfonylbenzoic acid in 120 ml of 
dioxane. Stirring is maintained for 18 hours and the solvent evaporated. 
The residue is taken up in water and extracted with ethyl acetate. After 
drying and evaporation, the expected product is obtained after 
purification on a silica column, using a dichloromethane/ethyl acetate 
(90/10) mixture as eluent. 
Melting point: 146.degree. C. 
Preparation C: 4-[2-(1-Oxa-2-oxo-3,8-diazaspiro[4.5]dec-8-yl)ethyl]benzoic 
acid 
100 mmol of 4-(2-chloroethyl)benzoic acid and 100 mmol of 
1-oxa-2-oxo-3,8-diazaspiro[4.5]decane are dissolved in 300 ml of methyl 
isobutyl ketone in the presence of 300 mmol of potassium carbonate and 
0.25 g of potassium iodide. The whole is maintained for 30 hours at 
reflux, with stirring, and then evaporated. The residue is taken up in 
water and washed with ethyl acetate. The aqueous phase is acidified with 
concentrated hydrochloric acid (pH=1), filtered and the expected product 
is fixed to resin, washed with water and eluted with 20% aqueous ammonia. 
After evaporation, the expected product is obtained after purification on 
a silica column, using an acetone/water (90/10) mixture as eluent. 
Melting point:&gt;250.degree. C. 
Preparation D: 4-[2-(4-Hydroxy-3,5-di-tert-butylphenylthio)ethyl]benzoic 
acid 
50 mmol of 4-hydroxy-3,5-di-tert-butylthiophenol, 50 mmol of 
4-(2-chloroethyl)benzoic acid and 100 ml of dimethylformamide are added, 
with stirring and under an inert atmosphere, to a solution containing 50 
mmol of sodium ethoxide in 200 ml of ethanol. The whole is heated at 
65.degree.-70.degree. C. for 8 hours. The ethanol is evaporated, the 
remaining phase taken up in ethyl acetate, filtered and evaporated. The 
expected product is obtained by purification of the residue by 
chromatography on silica gel, using a dichloromethane/dioxane (95/5) 
mixture as eluent. 
Melting point: 147.degree. C. 
Elemental microanalysis: 
______________________________________ 
C % H % S % 
______________________________________ 
calculated 71.47 7.82 8.29 
found 71.01 8.04 8.02 
______________________________________ 
Preparation E: 
4-[4-[2-(4-Hydroxy-3,5-di-tert-butylphenylthio)ethyl]benzoylaminosulfonyl] 
benzoic acid 
6 mmol of N-methylmorpholine are added to 6 mmol of the compound obtained 
in Preparation D in 70 ml of anhydrous tetrahydrofuran (THF) under an 
inert atmosphere and the whole is cooled to 5.degree. C. 6 mmol of 
isobutyl chloroformate in a solution of 10 ml of anhydrous THF are then 
slowly added. After returning to room temperature, the whole is stirred 
for 30 minutes. 6 mmol of 4-carbethoxyphenylsulfonamide in 25 ml of 
anhydrous THF are then slowly added and stirring is maintained for 18 
hours. After evaporation, the residue is taken up in ethyl acetate, washed 
with a saturated sodium bicarbonate solution, with 10% citric acid and 
with water, dried and evaporated. The ester thus obtained is purified by 
chromatography on a silica column, using a 
dichloromethane/methanol/aqueous ammonia (90/10/1) mixture as the elution 
solvent. The expected product is then obtained by saponification of the 
ester. 
Preparation F: 
4-[(2,5,7,8-Tetramethyl-6-hydroxychroman-2-yl)carbonylaminosulfonyl]benzoi 
c acid 
The expected product is obtained by using the same process as that 
described in Preparation A. 
Melting point: 128.degree. C. 
Preparation G: 4-[S-tert-Butyl-N-acetylcysteinylaminosulfonyl]benzoic acid 
The expected product is obtained by using the same process as that 
described in Preparation A. 
Melting point: 227.degree. C. 
Preparation H: 
4-[2-[(1,3-Thiazolidin-3-yl)carbonyl]benzoylaminosulfonyl]benzoic acid 
The expected product is obtained by using the same process as that 
described in Preparation A. 
Melting Doint:&gt;250.degree. C. 
Preparation I: 
4-[(4-Hydroxy-2-methyl-1,1-dioxo-1,2-benzothiazin-3-yl)carbonyl]benzoic 
acid 
Stage A: N-[(4-Carbethoxybenzoyl)methyl]saccharin 
45 mmol of ethyl 4-bromoacetylbenzoate in 120 ml of dimethylformamide and 
48 mmol of sodium saccharinate are maintained at 100.degree. C. for 180 
minutes. After evaporation of the solvent, taking up in 200 ml of water, 
extraction with dichloromethane, drying and evaporation, the residue is 
taken up in isopropyl ether, then filtered and taken to the expected 
product. 
Stage B: 3-(4-Carbethoxybenzoyl)-1,1-dioxo-1,2-benzothiazin-4-one 
The expected product is obtained from the compound described in Stage A 
according to the method described by H. Zinner et al. (J. O. C., 30, 
2241-2246, 1965). 
Melting point: 161.degree. C. 
Stage C: 3-(4-Carbethoxybenzoyl)-2-methyl-1,1-dioxo-1, 2-benzothiazin-4-one 
The expected product is obtained from the compound described in Stage B 
according to the method described by H. Zinner et al. (J. O. C., 30, 
2241-2246, 1965). 
Stage D: 
4-[(4-Hydroxy-2-methyl-1,1-dioxo-1,2-benzothiazine-3-yl)carbonyl]benzoic 
acid 
The expected product is obtained by saponification of 7 mmol of the 
compound obtained in the preceding stage in 30 ml of 0.5N sodium hydroxide 
solution and 15 ml of ethanol for 48 hours. After evaporation of the 
ethanol and acidification with 20 ml of 1N hydrochloric acid, the 
precipitate is filtered and washed with water. 
Melting point:&gt;260.degree. C. 
Preparation J: 4-[(Dicyclopropylmethyl)carbonylaminosulfonyl]benzoic acid 
The expected product is obtained by using the same process as that 
described in Preparation A, using dicyclohexylcarbodiimide as the coupling 
agent. 
Elemental microanalysis: 
______________________________________ 
C % H % N % S % 
______________________________________ 
calculated 55.72 5.30 4.33 9.92 
found 55.93 5.38 4.89 9.99 
______________________________________ 
Preparation K: 4-[(4-Chlorophenyl)ureidosulfonyl]benzoic acid 
The compound was obtained according to the technique described in 
Synthesis, 3, 221, 1990. 
Elemental microanalysis: 
______________________________________ 
C % H % N % S % 
______________________________________ 
calculated 50.20 3.95 7.32 8.38 
found 50.30 3.96 7.53 8.15 
______________________________________ 
Preparation L: 4-[(Adamant-1-yl)carbonylaminosulfonyl]benzoic acid 
25 mmol of the chloride of adamantane-1-carboxylic acid and 25 mmol of 
4-ethoxycarbonylbenzene- sulfonamide are maintained at reflux of 100 ml of 
anhydrous toluene in the presence of 3.45 ml of triethylamine for 20 
hours. After cooling, the toluene phase is washed with water, then with a 
NaHCO.sub.3 solution and then again with water. The ester thus obtained is 
purified by chromatography on a silica column, using a 
dichloromethane/acetone (98/2) mixture as eluent. The expected product is 
then obtained by saponification of the ester in a sodium hydroxide/ethanol 
mixture and is purified by chromatography on a silica column, using a 
dichloromethane/methanol (90/10) mixture as eluent. 
Preparation M: 
4-[(3,5-Di-tert-butyl-4-hydroxybenzyl)carbonylaminosulfonyl]benzoic acid 
The expected product was prepared according to the process described in 
Preparation J. 
Elemental microanalysis: 
______________________________________ 
C % H % N % S % 
______________________________________ 
calculated 61.73 6.53 3.13 7.16 
found 61.37 6.60 3.35 7.07 
______________________________________ 
Preparation N: 
4-[4-(2,6-Dimethyl-3,5-diethoxycarbonylpyrid-4-yl)benzoylaminosulfonyl]ben 
zoic acid 
The expected product was prepared according to the process described in 
Preparation J, using 1-(3-dimethylaminopropyl)-3ethylcarbodiimide as the 
coupling agent. 
Preparation O: 
4-{4-[2-(1-Oxa-2-oxo-3,8-diazaspiro[4.5]-dec-8-yl)ethyl]benzoylaminosulfon 
yl}-benzoic acid 
The expected product was prepared according to the process described in 
Preparation N from the compound described in Preparation C and 
4-ethoxycarbonylbenzene-sulfonamide. 
Preparation P: 
4-{4-Chloro-3-[(3,5-di-tert-butyl-4-hydroxybenzyl)carbonylaminosulfonyl]be 
nzoylaminosulfonyl}benzoic acid 
The expected product was obtained by coupling 
4-hydroxy-3,5-di-tert-butylphenylacetic acid with ethyl 
4-chloro-3-sulfamoylbenzoate according to the process described in 
Preparation J, saponification and then again coupling the compound 
obtained with 4-ethoxycarbonyl-benzenesulfonamide. 
Preparation Q: 4-[3,5-Di-tert-butyl-4-(3,5-di-tert- 
butyl-4-hydroxybenzoyloxy)benzoylaminosulfonyl]benzoic acid 
The expected product was obtained according to the process described in 
Preparation J. 
Elemental microanalysis: 
______________________________________ 
C % H % N % S % 
______________________________________ 
calculated 66.74 7.11 2.10 4.82 
found 66.38 7.48 2.23 5.20 
______________________________________ 
Preparation R: 
4-{[4-Methoxy-3-(3-ethoxy-2-hydroxypropoxy)benzoyl]aminosulfonyl}benzoic 
acid 
The expected product was prepared from 
4-methoxy-3-(2,3-epoxypropoxy)benzoic acid and 
4-ethoxycarbonylbenzenesulfonamide according to the process described in 
Preparation N. 
Mass spectrum: Chemical ionization/NH.sub.3 [M+H].sup.+ : m/z=454 
(theoretical mass M=453 ) 
Preparation S: 4-[(3,5-Di-tert-butyl-4-hydroxy)benzoylaminosulfonyl]benzoic 
acid 
The compound was obtained according to the process described in Preparation 
J. 
Preparation T: 
4-{4-[3,5-Di-tert-butyl-4-(ethoxymethoxy)benzyloxy]benzoylaminosulfonyl}be 
nzoic acid 
The compound was prepared according to the process described in Preparation 
J. 
Preparation U: 
4-[(3,5-Di-tert-butyl-4-hydroxyphenylthio)acetylaminosulfonyl]benzoic acid 
The compound was prepared according to the process described in Preparation 
J. 
Preparation V: 
4-{[3-(3,5-Di-tert-butyl-4-ethoxymethoxyphenyl)-trans-acryloyl]aminosulfon 
yl}benzoic acid 
The compound was prepared according to the process described in Preparation 
J. 
Elemental microanalysis: 
______________________________________ 
C % H % N % S % 
______________________________________ 
calculated 62.65 6.82 2.71 6.19 
found 62.46 7.25 2.93 6.56 
______________________________________

EXAMPLE 1 
4-(4-Chlorobenzoylaminosulfonyl)benzoyl-(S)-Val-(S)-Abo-(R,S)-Val-CF.sub.3 
Stage A: Boc-(S)-Val-(S)-Abo-OCH.sub.2 CH.sub.3 
By using the peptide coupling technique described by W. Konig and R. Geiger 
(Chem. Ber., 103, 788, 1970), 100 mmol of Boc-(S)-Val-OH and 100 mmol of 
3-(S)-carbethoxy-2-azabicyclo[2.2.2]octane are reacted. The expected 
product then obtained after purification on silica gel, using a 
dichloromethane/ethanol (97/3) mixture as eluent. 
Yield: 80% 
Elemental microanalysis: 
______________________________________ 
C % H % N % 
______________________________________ 
calculated 62.80 8.96 7.32 
found 62.42 8.78 7.26 
______________________________________ 
Stage B: Boc-(S)-Val-(S)-Abo-OH 
The expected product is obtained by saponification of the compound obtained 
in the preceding stage in a solution containing 150 ml of ethanol and 100 
ml of 1N sodium hydroxide solution for 72 hours. After evaporation of the 
ethanol and addition of 200 ml of water, washing the aqueous phase with 
ether and acidification of this by addition of citric acid, the compound 
precipitates and is washed with water and then with ether. 
Yield: 80% 
Melting point: 186.degree. C. 
Stage C: 
N-(Boc-(S)-Val-(S)-Abo)-1-isopropyl-2-hydroxy-3,3,3-trifluoropropylamine 
21 mmol of the compound obtained in the preceding stage and 21 mmol of 
1-isopropyl-2-hydroxy-3,3,3-trifluoropropylamine hydrochloride (described 
in J. Med. Chem., 33., 394-407, 1990) are coupled according to the peptide 
coupling technique described in Stage A. The expected product is obtained 
after purification on silica gel, using a dichloromethane/ethanol 
(97.5/2.5) mixture as eluent. 
Yield: 96% 
15 Elemental microanalysis: 
______________________________________ 
C % H % N % 
______________________________________ 
calculated 56.79 7.94 8.28 
found 56.65 7.74 8.48 
______________________________________ 
Stage D: Boc-(S)-Val-(S)-Abo-(R,S)-Val-CF.sub.3 
5 mmol of the compound obtained in the preceding stage are dissolved in 30 
ml of dimethyl sulfoxide and 30 ml of acetic anhydride. After stirring for 
20 hours at room temperature and addition of 75 ml of water, stirring is 
maintained for an additional hour. The whole is poured onto 150 ml of 
water and basified by addition of sodium bicarbonate. After extraction 
with dichloromethane, washing the organic phases with water, drying and 
evaporation, the expected product is obtained after purification on a 
silica column, using a dichloromethane/acetone (90/10 ) mixture as eluent. 
Yield: 65% 
Elemental microanalysis: 
______________________________________ 
C % H % N % 
______________________________________ 
calculated 57.02 7.58 8.31 
found 57.69 7.58 8.12 
______________________________________ 
Stage E: (S)-Val-(S)-Abo-(R,S)-Val-CF.sub.3 hydrochloride 
7 mmol of the product obtained in the preceding stage are deprotected by 
stirring, at room temperature, in 100 ml of a 3N hydrochloric acid 
solution in ethyl acetate. After evaporation of the solvent and addition 
of 100 ml of ether, the gel thus obtained is dried and taken to the 
expected product. 
Yield: 90% 
Stage F: 
4-(4-Chlorobenzoylaminosulfonyl)benzoyl-(S)-Val-(S)-(R,S)-Val-CF.sub.3 
1 mmol of the product obtained in the preceding stage and 1 mmol of 
4-(4-chlorobenzoylaminosulfonyl)-benzoic acid described in Preparation A 
are coupled according to the technique described by B. Castro et al. (Tet. 
Lett., 14, 1219-1222, 1975). After evaporation of the dimethylformamide, 
the residue is dissolved in 100 ml of ethyl acetate, washed with 50 ml of 
a saturated sodium bicarbonate solution and then with water. After drying 
and evaporation, the expected product is obtained by chromatography on 
silica gel, using a dichloromethane/methanol (90/10) mixture as eluent. 
Yield: 60% 
Mass spectrum: FAB-[M+H].sup.+ : m/z=727 (theoretical mass-.sup.35 
Cl-M=726) 
Examples 2 to 20 have been obtained by using the same process as that 
described for Example 1, using the corresponding starting materials. 
EXAMPLE 2 
4-{[3-(3-Azabicyclo[3.3.0]octane)ureido]sulpfonyl}benzoyl-(S)-Val-(S)-Abo-( 
R,S)-Val-CF.sub.3 
Mass spectrum: FAB-[M+H].sup.+ : m/z=741 (theoretical mass M=740) 
EXAMPLE 3 
4-(4-Chlorophenylsulfonylaminocarbonyl)benzoyl-(S)-Val-(S)-Abo-(R,S)-Val-CF 
.sub.3 
Mass spectrum: FAB-[M-H].sup.- : m/z=725 (theoretical mass-.sup.35 
Cl-M=726) 
EXAMPLE 4 
4-(4-Chlorophenylsulfonylaminocarbonyl)benzoyl-(S)-Val-(S)-Abo-Val-CF.sub.3 
, .alpha. isomer 
Mass spectrum: FAB-[M-H].sup.- : m/z=725 (theoretical mass-.sup.35 
Cl-M=726) 
EXAMPLE 5 
4-(4-Chlorophenylsulfonylaminocarbonyl)benzoyl-(S)-Val-(S)-Abo-Val-CF.sub.3 
, .beta. isomer 
Mass spectrum: FAB-[M-H].sup.- : m/z=725 (theoretical mass-.sup.35 
Cl-M=726) 
The .alpha. and .beta. isomers of the compound of Example 3 are separated 
by C.sub.18 reverse phase preparative liquid chromatography, using a 0.05M 
NaHCO.sub.3 -water/acetonitrile (80/20) mixture as eluent. The eluates are 
concentrated, acidified with 10% citric acid, extracted with 
dichloromethane, washed with water, dried and evaporated. The .alpha. and 
.beta. isomers are named thus by the order of departure from the column. 
EXAMPLE 6 
4-(4-Chlorophenylsulfonylaminocarbonyl)benzoyl-(S)-Val-(2S,3aS,7aS)-Phi-(R, 
S)-Val-CF.sub.3 
Mass spectrum: FAB-[M-H].sup.- : m/z=739 (theoretical mass-.sup.35 
Cl-M=740) 
EXAMPLE 7 
4-(4-Chlorophenylsulfonylaminocarbonyl)benzoyl-(S)-Val-(2S,3aS,7aS)-Phi-Val 
-CF.sub.3, .alpha. isomer 
Mass spectrum: FAB-[M-H].sup.- : m/z=739 (theoretical mass-.sup.35 
Cl-M=740) 
EXAMPLE 8 
4-(4-Chlorophenylsulfonylaminocarbonyl)benzoyl-(S)-Val-(2S,3aS,7aS)-Phi-Val 
-CF.sub.3, .beta. isomer 
Mass spectrum: FAB-[M-H].sup.- : m/z=739 (theoretical mass-.sup.35 
Cl-M=740) 
The .alpha. and .beta. isomers of the compound of Example 6 are separated 
according to the same technique as that described for Examples and 5. The 
elution solvent is a mixture: 0.05M NaHCO.sub.3 -water/acetonitrile: 
75/25. 
EXAMPLE 9 
4-[(R,S)-2-Oxotetrahydrothiophen-3-yl)aminosulfonyl]-benzoyl-(S)-Val-(S)-Ab 
o-(R,S)-Val-CF.sub.3 
Mass spectrum: FAB-[M-H].sup.- : m/z=687 (theoretical mass-M=688) 
EXAMPLE 10 
4-[(2-Oxotetrahydrothiophen-3-yl)aminosulfonyl]benzoyl-(S)-Val-(S)-Abo-Val- 
CF.sub.3, mixture of .alpha. isomers 
Mass spectrum: FAB-[M-H].sup.- : m/z=687 (theoretical mass-M=688) 
EXAMPLE 11 
4-[(2-Oxotetrahydrothiophen-3-yl)aminosulfonyl]benzoyl-(S)-Val-(S)-Abo-Val- 
CF.sub.3, mixture of .beta. isomers 
Mass spectrum: FAB-[M-H].sup.- : m/z=687 (theoretical mass-M=688) 
The expected products are obtained by using the product described in 
Preparation B in Stage F. The compound of Example is a mixture of 4 
isomers, two pairs of which are separated according to the technique 
described for Examples 4 and 5. The elution solvent is a mixture: 0.05M 
NaHCO.sub.3 -water/acetonitrile: 65/35. 
EXAMPLE 12 
4-[2-(1-Oxa-2-oxo-3,8-diazaspiro[4.5]dec-8-yl)ethyl]benzoyl-(S)-Val-(S)-Abo 
-(R,S)-Val- CF.sub.3 
The expected product is obtained by using the product described in 
Preparation C in Stage F. 
Mass spectrum: FAB-[M+H].sup.+ : m/z=692 (theoretical mass-M=691) 
EXAMPLE 13 
4-[4-[2-(1-Oxa-2-oxo-3,8-diazaspiro[4.5]dec-8-yl)ethyl]benzoylaminosulfonyl 
]benzoyl-(S)-Val-(S)-Abo-(R,S)-Val-CF.sub.3 
EXAMPLE 14 
4-[(4-Hydroxy-3,5-di-tert-butylphenylacetyl)aminosulfonyl]benzoyl-(S)-Val-( 
S)-Abo-(R,S)-Val-CF.sub.3 
Elemental microanalysi: 
______________________________________ 
C % H % N % S % 
______________________________________ 
calculated 60.42 6.88 6.71 3.84 
found 60.96 7.20 7.25 3.38 
______________________________________ 
EXAMPLE 15 
4-[2-(4-Hydroxy-3,5-di-tert-butylphenylthio)ethyl]benzoyl-(S)-Val-(S)-Abo-( 
R,S)-Val-CF.sub.3 
The expected product is obtained by using the product described in 
Preparation D in Stage F. 
Elemental microanalysis: 
______________________________________ 
C % H % N % S % 
______________________________________ 
calculated 65.18 7.55 5.43 4.14 
found 65.28 7.77 5.23 4.03 
______________________________________ 
EXAMPLE 16 
4-[4-[2-(4-Hydroxy-3,5-di-tert-butylphenylthio)ethyl]benzoylaminosulfonyl]b 
enzoyl-(S)-Val-(S)-Abo-(R,S)-Val-CF.sub.3 
The expected product is obtained by using the product described in 
Preparation E in Stage F. 
EXAMPLE 17 
4-[(2,5,7,8-Tetramethyl-6-hydroxychroman-2-yl)carbonylaminosulfonyl]benzoyl 
-(S)-Val-(S)-Abo-(R,S)-Val-CF.sub.3 
The expected product is obtained by using the product described in 
Preparation F in Stage F. 
Elemental microanalysis: 
______________________________________ 
C % H % N % S % 
______________________________________ 
calculated 58.52 6.26 6.82 3.91 
found 58.50 6.51 7.04 3.65 
______________________________________ 
EXAMPLE 18 
4-[S-tert-Butyl-N-acetyl-(S)-cysteinylaminosulfonyl]benzoyl-(S)-Val-(S)-Abo 
-(R,S)-Val- CF.sub.3 
The expected product is obtained by using the product described in 
Preparation G in Stage F. 
Elemental microanalysis: 
______________________________________ 
C % H % N % S % 
______________________________________ 
calculated 53.22 6.38 8.87 8.12 
found 53.22 6.28 8.57 8.38 
______________________________________ 
EXAMPLE 19 
4-[2-[(1,3-Thiazolidin-3-yl)carbonyl]benzoylaminosulfonyl]benzoyl-(S)-Val-( 
S)-Abo-(R,S)-Val-CF.sub.3 
The expected product is obtained by using the product described in 
Preparation H in Stage F. 
EXAMPLE 20 
4-[(4-Hydroxy-2-methyl-1,1-dioxo-1,2-benzothiazin-3-yl)carbonyl]benzoyl-(S) 
-Val-(S)-Abo-(R,S)-Val-CF.sub.3 
The expected product is obtained by using the product described in 
Preparation I in Stage F. 
Elemental microanalysis: 
______________________________________ 
C % H % N % S % 
______________________________________ 
calculated 57.90 5.53 7.50 4.29 
found 57.53 5.53 7.48 4.30 
______________________________________ 
EXAMPLE 21 
4-[(4-Hydroxy-1,1-dioxo-(2H)-1,2-benzothiazin-3-yl)carbonyl]benzoyl-(S)-Val 
-(S)-Abo-(R,S)-Val-CF.sub.3 
The expected product is obtained by using, in Stage F, the product 
synthesized by saponification of the compound described in Preparation I. 
Mass spectrum: FAB-[M+H].sup.+ : m/z=733 (theoretical mass-M=732) 
EXAMPLE 22 
4-(4-Chlorophenylsulfonylaminocarbonyl)benzoyl-(S)-Val-(1S,3S,4R)-Abh-(R,S) 
-Val-CF.sub.3 
Stage A: 
N-[(1S,3S,4R)-Abh]-2-hydroxy-1-isopropyl-3,3,3-trifluoropropylamine hydroc 
hloride 
21 mmol of Boc-(1S,3S,4R)-Abh-OH are coupled, according to the peptide 
coupling technique of W. Konig and R. Geiger, with 21 mmol of 
2-hydroxy-1-isopropyl-3,3,3-trifluoropropylamine hydrochloride. The 
expected product is obtained after purification on a silica column, using 
a dichloromethane/methanol (97/3) mixture as eluent. 
Stage B: 
N-[Boc-(S)-Val-(1S,3S,4R)-Abh]-2-hydroxy-1-sopropyl-3,3,3-trifluoropropyla 
mine 
The expected product is obtained by coupling 12 mmol of the compound 
described in Stage A with 12 mmol of Boc-(S)-Val-OH according to the same 
peptide coupling technique, and is purified by chromatography on a silica 
column, using a dichloromethane/ethanol (95/5) mixture as elution solvent. 
Stage C: Boc-(S)-Val-(1S,3S,4R)-Abh-(R,S)-Val-CF.sub.3 
The expected product is obtained by oxidation of the compound described in 
Stage B, according to the same process as that described in Stage D of 
Example 1, and is purified by chromatography on a silica column, using a 
dichloromethane/acetone (93/7) mixture as eluent. 
Stage D: (S)-Val-(1S,3S,4R)-Abh-(R,S)-Val-CF.sub.3 hydrochloride 
The expected product is obtained by deprotecting the compound described in 
Stage C according to the same technique as that described in Stage E of 
Example 1. 
Stage E: 
4-(4-Chlorophenylsulfonylaminocarbonyl)benzoyl-(S)-Val-(1S,3S,4R)-Abh-(R,S 
)-Val-CF.sub.3 
The expected product is obtained according to the process described in 
Stage F of Example 1 by reacting the compound described in Stage D with 
4-(4-chlorophenylsulfonylaminocarbonyl)benzoic acid and is purified by 
chromatography on a silica column, using a dichloromethane/methanol 
(90/10) mixture and eluent. 
Mass spectrum: FAB-[M+H].sup.+ : m/z=713 (theoretical mass-.sup.35 
Cl-M=712) 
EXAMPLE 23 
4-[2-[(1,3-Thiazolidin-3-yl)carbonyl]benzoylaminosulfonyl]benzoyl-(S)-Val-( 
lS,3S,4R)-Abh-(R,S)-Val-CF.sub.3 
The expected product is obtained according to the process described in 
Example 22 by using the product described in Preparation H in Stage E. 
Mass spectrum: FAB-[M+H].sup.+ : m/z=794 (theoretical mass-M=793) 
Examples 24 to 39 were obtained according to the same process as that 
described in Example 1, using the corresponding starting materials. 
EXAMPLE 24 
4-[(Dicyclopropylmethyl)carbonylaminosulfonyl]benzoyl-(S)-Val-(S)-Abo-(R,S) 
-Val-CF.sub.3 
The expected product is obtained by using the product described in 
Preparation J in Stage F. 
Elemental microanalysis: 
______________________________________ 
C % H % N % S % 
______________________________________ 
calculated 57.45 6.38 7.88 4.51 
found 57.57 6.75 7.91 4.32 
______________________________________ 
EXAMPLE 25 
4-(4-Chlorophenylureidosulfonyl)benzoyl-(S)-Val-(S)-Abo-(R,S)-Val-CF.sub.3 
The expected product is obtained by using the product described in 
Preparation K in Stage F. 
Elemental microanalysis: 
______________________________________ 
C % H % N % Cl % S % 
______________________________________ 
calculated 
53.40 5.30 9.44 4.78 4.32 
found 53.86 5.28 9.55 4.22 4.77 
______________________________________ 
EXAMPLE 26 
4-(4-Chlorophenylureidosulfonyl)benzoyl-(S)-Val-(2S,3aS,7aS)-Phi-(R,S)-Val- 
CF.sub.3 
The expected product is obtained by using the product described in 
Preparation K in Stage F. 
Elemental microanalysis: 
______________________________________ 
C % H % N % Cl % S % 
______________________________________ 
calculated 
51.54 5.72 8.84 4.47 4.05 
found 51.74 5.85 8.53 4.09 3.65 
______________________________________ 
EXAMPLE 27 
4-[(Dicyclopropylmethyl)carbonylaminosulfonyl]benzoyl-(S)-Val-(2S,3aS,7aS)- 
Phi-(R,S)-Val-CF.sub.3 
The expected product is obtained by using the product described in 
Preparation J in Stage F. 
Elemental microanalysis: 
______________________________________ 
C % H % N % S % 
______________________________________ 
calculated 58.00 6.54 7.73 4.42 
found 58.20 6.91 7.95 4.38 
______________________________________ 
EXAMPLE 28 
4-[(Adamant-1-yl)carbonylaminosulfonyl]benzoyl-(S)-Val-(S)-Abo-(R,S)-Val-CF 
.sub.3 
The expected product is obtained by using the product described in 
Preparation L in Stage F. 
Elemental microanalysis: 
______________________________________ 
C % H % N % S % 
______________________________________ 
calculated 59.18 6.58 7.46 4.26 
found 59.18 7.04 8.05 4.10 
______________________________________ 
EXAMPLE 29 
4-[(3,5-Di-tert-butyl-4-hydroxybenzyl)carbonylaminosulfonyl]benzoyl-(S)-Val 
-(2S,3aS,7aS)-Phi-(R,S)-Val-CF.sub.3 
The expected product is obtained by using the product described in 
Preparation M in Stage F. 
Elemental microanalysis: 
______________________________________ 
C % H % N % S % 
______________________________________ 
calculated 60.83 7.00 6.60 3.78 
found 60.67 7.33 6.97 3.85 
______________________________________ 
EXAMPLE 30 
4-[4-(2,6-Dimethyl-3,5-diethoxycarbonyl-1,4-dihydropyrid-4-yl)benzoylaminos 
ulfonyl]benzoyl-(S)-Val-(S)-Abo-(R,S)-Val-CF.sub.3 
The expected product is obtained by using the product described in 
Preparation N in Stage F. 
Elemental microanalysis: 
______________________________________ 
C % H % N % S % 
______________________________________ 
calculated 58.53 5.98 7.42 3.40 
found 57.86 5.94 7.12 3.30 
______________________________________ 
EXAMPLE 31 
4-{4-[2-(3,5-Di-tert-butyl-4-hydroxyphenylthio)ethyl]benzoylaminosulfonyl}b 
enzoyl-(S)-Val-(2S,3aS,7aS)-Phi-(R,S)-Val-CF.sub.3 
The expected product is obtained by using the product described in 
Preparation E in Stage F. 
Elemental microanalysis: 
______________________________________ 
C % H % N % S % 
______________________________________ 
calculated 61.84 6.75 5.77 6.60 
found 61.73 7.04 5.49 5.49 
______________________________________ 
EXAMPLE 32 
4-{4-[2-(1-Oxa-2-oxo-3,8-diazaspiro[4.5]dec-8-yl)ethyl]benzoylaminosulfonyl 
}benzoyl-(S)-Val-(2S,3aS,7aS)-Phi-(R,S)-Val-CF.sub.3 
The expected product is obtained by using the product described in 
Preparation O in Stage F. 
Mass spectrum: FAB-[M].sup.+ : m/z=888 (theoretical mass: M=888 ) 
EXAMPLE 33 
4-{[4-Chloro-3-[(3,5-di-tert-butyl-4-hydroxybenzyl)carbonylaminosulfonyl]be 
nzoyl]aminosulfonyl}benzoyl-(S)-Val-(2S,3aS,7aS)-Phi-(R,S)-Val-CF.sub.3 
The expected product is obtained by using the product described in 
Preparation P in Stage F. 
Elemental microanalysis: 
______________________________________ 
C % H % N % S % 
______________________________________ 
calculated 56.30 5.95 6.57 6.01 
found 56.16 6.45 6.67 6.08 
______________________________________ 
EXAMPLE 34 
4-{[3,5-Di-tert-butyl-4-(3,5-di-tert-butyl-4-hydroxybenzoyloxy)benzoyl]amin 
osulfonyl}-benzoyl-(S)-Val-(2S,3aS,7aS)-Phi-(R,S)-Val-CF.sub.3 
The expected product is obtained by using the product described in 
Preparation Q in Stage F. 
Elemental microanalysis: 
______________________________________ 
C % H % N % S % 
______________________________________ 
calculated 64.14 7.27 5.25 3.00 
found 63.81 7.32 5.39 3.73 
______________________________________ 
EXAMPLE 35 
4-{[4-Methoxy-3-(3-ethoxy-2-hydroxypropoxy)-benzoyl]aminosulfonyl 
}benzoyl-(S)-Val-(2S,3aS,7aS)-Phi-(R,S)-Val-CF.sub.3 
The expected product is obtained by using the product described in 
Preparation R in Stage F. 
Mass spectrum: FAB-[M-H].sup.- : m/z=853 (theoretical mass: M=854) 
EXAMPLE 36 
4-[(3,5-Di-tert-butyl-4-hydroxybenzoyl)aminosulfonyl]benzoyl-(S)-Val-(2S,3a 
S,7aS)Phi-(R,S)-Val-CF.sub.3 
The expected product is obtained by using the product described in 
Preparation S in Stage F. 
Elemental microanalysis: 
______________________________________ 
C % H % N % S % 
______________________________________ 
calculated 60.42 6.88 6.71 3.84 
found 60.84 7.28 6.92 4.21 
______________________________________ 
EXAMPLE 37 
4-{4-[3,5-Di-tert-butyl-4-(ethoxymethoxy)benzyloxy]benzoylaminosulfonyl}ben 
zoyl-(S)-Val-(2S,3aS,7aS)-Phi-(R,S)-Val-CF.sub.3 
The expected product is obtained by using the product described in 
Preparation T in Stage F. 
Elemental microanalysis: 
______________________________________ 
C % H % N % S % 
______________________________________ 
calculated 62.51 6.96 5.61 3.21 
found 62.17 7.37 5.74 3.23 
______________________________________ 
EXAMPLE 38 
4-[(3,5-Di-tert-butyl-4-hydroxyphenylthio)acetylaminosulfonyl]benzoyl-(S)-V 
al-(2S,3aS,7aS)-Phi-(R,S)-Val-CF.sub.3 
The expected product is obtained by using the product described in 
Preparation U in Stage F. 
Elemental microanalysis: 
______________________________________ 
C % H % N % S % 
______________________________________ 
calculated 58.62 6.75 6.36 7.28 
found 58.79 7.12 6.02 7.93 
______________________________________ 
EXAMPLE 39 
4-{[3-(3,5-Di-tert-butyl-4-ethoxymethoxyphenyl)-trans-acryloyl]aminosulfony 
l}benzoyl-(S)-Val-(2S,3aS,7aS)-Phi-(R,S)-Val-CF.sub.3 
The expected product is obtained by using the product described in 
Preparation V in Stage F. 
Elemental microanalysis: 
______________________________________ 
C % H % N % S % 
______________________________________ 
calculated 61.42 7.13 6.10 3.49 
found 61.22 7.10 6.10 3.43 
______________________________________ 
EXAMPLE 40 
4-{[3-(3,5-Di-tert-butyl-4-hydroxyphenyl)trans-acryloyl]aminosulfonyl}benzo 
yl-(S)-Val-(2S,3aS,7aS)-Phi-(R,S)-Val-CF.sub.3 
The expected product was obtained after hydrolysis in dioxane/hydrochloric 
acid medium of the compound described in Example 39 and purification by 
chromatography on a silica column, using a dichloromethane/methanol (95/5) 
mixture as eluent. 
Elemental microanalysis: 
______________________________________ 
C % H % N % S % 
______________________________________ 
calculated 61.38 7.22 6.42 4.18 
found 61.12 6.91 6.51 3.72 
______________________________________ 
EXAMPLE 41 
4-(4-Chlorophenylsulfonylaminocarbonyl)benzoyl-[N-(3,5-di-tert-butyl-4-hydr 
oxyphenyl)acetyl]-(S)-Lys-(2S,3aS,7aS)-Phi-(R,S)-Val-CF.sub.3 
Stage A: 4-(4-Chlorophenylsulfonylaminocarbonyl)benzoyl-(S)-Lys-OCH.sub.3 
4-(4-Chlorophenylsulfonylaminocarbonyl)benzoic acid is coupled with the 
methyl ester of (S)-lysine, protected by a tert-butoxy group on its side 
amine functional group, by using BOP as a coupling reagent in the presence 
of diethylamine in dimethylformamide. The expected product is then 
deprotected by acid hydrolysis with the aid of hydrobromic acid in acetic 
acid medium and is purified by chromatography on a silica column, using a 
dichloromethane/methanol (60/40) mixture as eluent. 
Stage B: 
4-(4-Chlorophenylsulfonylaminocarbonyl)benzoyl-[N-(3,5-di-tert-butyl-4-hyd 
roxyphenyl)acetyl]-(S)-Lys-OH 
The compound obtained in the preceding stage is coupled, according to the 
technique of W. Konig and R. Geiger (Chem. Ber., 103, 788, 1970) with 
3,5-di-tert-butyl-4-hydroxyphenylacetic acid. The expected product, in the 
form of a methyl ester, is purified by chromatography on a silica column, 
using a dichloromethane/methanol (90/10) mixture as eluent, and is then 
saponified with a sodium hydroxide/ethanol mixture. 
Stage C: 
4-(4-Chlorophenylsulfonylaminocarbonyl)benzoyl-[N-(3,5-di-tert-butyl-4-hyd 
roxyphenyl)acetyl]-(S)-Lys-(2S, 3aS, 7 aS)-Phi-(R,S)-Val-CF.sub.3 
The product obtained in the preceding stage is coupled with 
(S)-Phi-(R,S)-Val-CF.sub.3, according to the technique described in the 
preceding stage, and taken to the expected product which is purified by 
chromatography on a silica column, using a dichloromethane/methanol (90/10 
) mixture as eluent. 
Elemental microanalysis: 
______________________________________ 
C % H % N % S % 
______________________________________ 
calculated 60.26 6.44 6.89 3.15 
found 59.37 6.59 6.43 2.77 
______________________________________ 
Examples 42 and 43 were obtained according to the process described for 
Example 41. 
EXAMPLE 42 
4-(4-Chlorophenylsulfonylaminocarbonyl)benzoyl-[N-(3,5-di-tert-butyl-4-hydr 
oxyphenylthio)butyroyl]-(S)-Lys-(2S,3aS,7aS)-Phi-(R,S)-Val-CF.sub.3 
Mass spectrum: FAB: [M+H].sup.+ : m/z=1076 (theoretical mass: M=1075) 
EXAMPLE 43 
4-(4-Chlorophenylsulfonylaminocarbonyl)benzoyl-[S-(3,5-di-tert-butyl-4-hydr 
oxy-phenyl)]-(S)-Cys-(2S,3aS,7aS)-Phi-(R,S)-Val-CF.sub.3 
Mass spectrum: FAB: [M+H].sup.+ : m/z=949 (theoretical mass: M=948) 
EXAMPLE 44 
##STR26## 
The expected product was obtained by hydrolysis of the compound of Example 
18 in a dioxane/water (80/20) mixture in the presence of hydrogen sulfide 
and of mercuric acetate. 
After filtration, the filtrate is evaporated and taken to the expected 
product. 
Elemental microanalysis: 
______________________________________ 
C % H % N % S % 
______________________________________ 
calculated 50.81 5.64 9.56 8.75 
found 51.42 5.58 9.50 8.32 
______________________________________ 
PHARMACOLOGICAL STUDY OF THE DERIVATIVES OF THE INVENTION 
EXAMPLE 45 
Inhibitory activity for human leucocyte elastase in vitro 
The strength of the compounds of the invention is determined by the level 
of inhibition of the action of human leucocyte elastase on a low molecular 
weight peptide substrate according to the technique described by B. M. 
Ashe et al., (J. Biol. Chem., 256, 11603-11606, 1981). This activity is 
measured by following the kinetics of hydrolysis of the substrate which 
leads to the release of paranitroaniline which absorbs at a wavelength of 
410 nm. 
Reagent: 
Enzyme: Human sputum leucocyte elastase (Elastin Products Co.) solubilized 
at 1000 uI/ml of distilled water, and frozen in aliquots of 50 .mu.l at 
-20.degree. C. 
Substrate: methoxysuccinyl-L-alanyl-L-alanyl-propyl-valine paranitroanilide 
(Sigma chimie). 
Buffer: Tris 0.1M; NaCl 0.5M; pH=7.8. 
Equipment: 
Spectrophotometer thermostatically controlled at 37.degree. C., equipped 
with a cell changer. 
1 ml polystyrene cell. 
Dry waterbath adjusted to 37.degree. C. 
Procedure: 
The following are introduced into a cell: 
970 .mu.l of buffer 
10 .mu.l of the product to be tested or of the solvent 
(concentrated.times.100) 
10 .mu.l of human sputum leucocyte elastase diluted to 1/10th 
Stirring and incubation for 15 min at 37.degree. C. 
The reaction is started by addition of 10 .mu.l of substrate. 
The cell is introduced into the spectro- photometer, and the optical 
density is recorded as a function of time at 410 nm at 37.degree. C. 
The initial rate is measured for each product concentration (or solvent 
control) studied. 
Percentages of inhibition with respect to the solvent control are 
calculated: 
EQU Percentage of inhibition=100.times.(control rate-rate of the product 
tested/control rate). 
The inhibitory concentrations 50 (IC.sub.50) are calculated from the 
percentages of inhibition by simple linear regression. 
The results obtained in this test are collated below: 
______________________________________ 
Example IC.sub.50 (nM) 
Example IC.sub.50 (nM) 
______________________________________ 
1 24 28 47 
2 25 29 38 
4 15 30 41 
6 27 31 40 
14 30 32 42 
18 19 33 34 
22 19 35 37 
23 30 36 36 
24 25 40 38 
25 34 41 70 
26 31 44 14 
27 29 
______________________________________ 
EXAMPLE 46 
In vivo inhibitory activity: model of acute hemorrhagic edema in hamsters 
Tracheal instillation in hamsters of a purified preparation of human sputum 
leucocyte elastase leads to acute hemorrhaging which can be quantified by 
measuring the concentration of hemoglobin in the broncho-alveolar washing 
3 hours after the elastase instillation. 
The study is carried out on male hamsters weighing 120 to 100 g (Syrian 
Golden, n=10 per batch). The animals are anesthetized with pentobarbital 
at a dose of 40 mg/kg intraperitoneally. 
The hamsters are anesthetized and the trachea is exposed surgically. The 
products to be tested are administered with the aid of a needle directly 
into the trachea in a volume of 0.1 ml at a dose of 10 .mu.g. Human sputum 
leucocyte elastase is administered, 3 hours after administration of the 
product, intratracheally at a dose of 50 units per animal at a volume of 
0.2 ml. 
The animals are sacrificed with the aid of a lethal dose of pentobarbital 3 
hours after instillation of elastase and a broncho-alveolar washing with 
physiological serum is carried out. The degree of hemorrhaging is 
quantified by a colorimetric method which makes it possible to 
quantitatively determine the hemoglobin concentration (Boehringer 
hemoglobin test combination). 
The results are expressed in percentage of inhibition of hemorrhaging. 
The compounds of the present invention are effective inhibitors of human 
leucocyte elastase which prevent or reduce the hemorrhaging induced by 
intratracheal instillation of human leucocyte elastase. The results 
obtained in this test are collated below: 
______________________________________ 
Inhibition Inhibition 
Example (%) Example (%) 
______________________________________ 
1 67 27 38 
2 57 28 57 
4 70 29 63 
6 77 30 56 
14 28 31 20 
18 38 32 42 
22 65 33 37 
23 61 35 47 
24 46 36 53 
25 63 40 40 
26 64 41 41 
44 44 
______________________________________ 
EXAMPLE 47 
Study of lipid peroxidation 
The study was carried out on rat hepatic microsomes in the presence of 
Fe.sup.3+ (100 .mu.M) and of ascorbate (100 .mu.M). Quantitative 
determination of malondialdehyde (MDA) is carried out by the 
thiobarbituric acid method (spectrophotometry .lambda.=532 nm) according 
to the technique described by N. Paya et al. (Biochem. Pharmacol., Vol. 
44, No. 2, p. 205-214, 1992). 
The products of the invention have an inhibitory activity of between 
10.sup.-6 M and 10.sup.-5 M on lipid peroxidation in the system studied. 
EXAMPLE 48 
Pharmaceutical composition 
Preparation formula for 1000 tablets each containing 10 mg of active 
material 
Compound of Example 6 . . . 10 g 
Hydroxypropyl cellulose . . . 2 g 
Wheat starch . . . 10 g 
Lactose . . . 100 g 
Magnesium stearate . . . 3 g 
Talc . . . 3 g