CH659475A5 - SUBSTITUTED PEPTIDE COMPOUNDS AND PHARMACEUTICAL COMPOSITIONS CONTAINING THE SAME. - Google Patents

Description

The invention relates to substituted peptide compounds having the formula (I) below and to the salts of these compounds.

o r rt o

II I I II

r3-ch-c-ch2-n — ch-c-x I (L)

nh I

c = o I

R2

where X is an amino acid or an imino acid of formula:

- (CH2) m-cycloalkyl,

O yRl

45

(I)

He / 15

_0 — C — N ', lower alkyl-O,

-o- (ch2)

50

(R13} p '

H, C CH_ 2 I I 2

CH_ / ^

H2? I 8

CH

st cH_

i2

a 1- or 2-naphthyloxy of formula:

-o- (ch_) Z m

-N C-COOR- / -N C-COOR ,. / -N - C-COOR- /

I (L) 6 '-

(L) 6

, (l) "'6 H M

lower alkyl-S,

-s- (ch2)

RioN * / R10 H2Ç f2

neither nor:

or a 1- or 2-naphthylthio having the formula:

65

■ N C-COOR, -N C-COOR- / -N C-COOR- /

.... 6 "(L) 6. ... c

(L)

Î (W 6

H

° / R15

Rg is a keto, a halogen, —O — C — N,

Laugh

5,659,475

"(CH2> B- @ / - (CH2) m-cycloaIkyl. - (CHj) £ -fTj),

-0- (ch2)

a lower alkyl-O,

- (ch,)

tR13 * p a 1- or 2-naphthyloxy having the formula:

<ê-

-0- (ch_) -2 m

'OI04- (r14) p.

a lower S-alkyl,

Rs is hydrogen, lower alkyl,

-s- <ch2ib or a 1- or 2-naphthylthio of formula:

-s- (ch-) - • Z ni

(R14> p-

R13 is hydrogen, lower alkyl of 1 to 4 carbon atoms, lower alkoxy of 1 to 4 carbon atoms, lower alkyl thio of 1 to 4 carbon atoms, chloro, bromo, fluoro, trifluoromethyl, hydroxy , phenyl, phenoxy, phenylthio or phenyl-methyl.

R14 is hydrogen, lower alkyl of 1 to 4 carbon atoms, lower alkoxy of 1 to 4 carbon atoms, lower alkyl thio of 1 to 4 carbon atoms, chloro, bromo, fluoro, trifluoromethyl or a hydroxy.

m is zero, one, two, three or four.

p is one, two or three, provided that p is greater than one, only if R13, or R14, is hydrogen, methyl, methoxy, chloro or fluoro.

Rls is hydrogen or lower alkyl of 1 to 4 carbon atoms.

Y is oxygen or sulfur.

R, 6 is a lower alkyl of 1 to 4 carbon atoms,

- (CVm

2.m

(R13> p

- (CH2) F © - (CH2) ^ Q ^ - ° H '

(CH.)

OH

20

(CH2)

—I N,

ïitjj

R9 is a keto or - (each) - 2 m

R10 is halogen or - Y — Ri6.

Ru, R'11, Rj2 and R'I2 are chosen independently, from hydrogen and lower alkyl or from R'llt R12 and R'12 are hydrogen, and RX1 is

NOT

I

H

- (CH,) r - NH ,, - (CH:) r - SH, - (CH:) r-S-lower alkyl,

- (CH2) r - NH - C

.NH

s \ h,

O

- (CH;) r —C —NH ,.

r is an integer from 1 to 4.

Rlg is lower alkyl, benzyl or phenethyl.

R20 is hydrogen, lower alkyl, benzyl or phenethyl. R is hydrogen, lower alkyl, cycloalkyl.

- (CH2) r-

- (CH2) 2 - NH2, - (CH,) 3 —NH2.

- (CH2) 4-NH2, - (CH2) 2-OH, - (CH2) 3-OH, - (CH2) 4-0H, - (CH2) 2-SH, - (CH2) 3 —SH, or - (CH2) 4-SH.

R, is hydrogen, lower alkyl, halo-substituted lower alkyl (CH,) r

. - (CH,) r -

-OH,

- <CH2'r

50

"(CH.)

-NOT

H

2'r F "lj '

not

I

H

55 - (CH,) r —NH ,, - (CH,) r —SH, - (CH,) r —OH, - (CH:) r-S-lower alkyl,

- (CH,) r-NH-C;

>

\

NH

O

there

or - (CH,) r —C —NH,

NH,

or the R16 groups joined to complete an unsubstituted 5 or 6 membered nucleus, or said nucleus in which one or more of the carbon atoms have a lower alkyl of 1 to 4 carbon atoms or a substituent di (lower alkyl of 1 to 4 carbon atoms).

R4 is hydrogen, lower alkyl,

provided that R is hydrogen, only if R is other than hydrogen.

"'" -'CV ..- ©. - <CH2'; rO.

"Wp * s

659475

- <CB2>, ï-fri). o „- (CH-.ìj-fo).

R3 is hydrogen, lower alkyl,

-'ca2 '£ - @ 1 - (CH2'rirfQ!'

Vf '

- (cV.rfÇ) '- <CH2>, r £ °)'

a halo-substituted lower alkyl, - (CH2) m-cycloalkyl,

- <CH2lr (o ^ -0H '-' ^ 'qrTo)'

H

- (CH2) r — OH, -ICHj) ^ ",

I

h

- (CH2) r — NH2, - (CH2) r — SH, - (CH2) t-S-lower alkyl,

>

, NH

O

The term "halo-substituted lower alkyl", or "halogen-substituted", refers to the aforementioned alkyl groups, in which one or more hydrogens are replaced by chloro, bromo or fluoro groups, for example, trifluoromethyl, which is preferably used, pentafluoroethyl, 2,2,2-trichloroethyl, chlo-romethyl, bromomethyl, etc.

The symbols:

/NOT

and z m

I

NS ^

z m

- (CB2'îT

O

mean that the alkylene bridge is linked to an available carbon atom.

The compounds of formula (I), where R is hydrogen, can be prepared by transforming a carboxymethyl peptide ester of formula:

O

I II

ho-c-ch2-n-ch-c-x (l)

(ii)

in its acid chloride, then reacting the latter with an oxazolone of formula:

n r2-c

c-r

(III)

- (ch2) t — nh — c, or - (ch2) r — c — nh2

nh2

where m, r14, p and r are defined as above.

r6 is hydrogen, lower alkyl, benzyl, benzhy-drile,

o r21o

II I II

—Çh — o — c — rj8, -ç-ç-0-r23, -ch- (ch-oh) 2,

Rl 7 1 ^ -22 1 ^ -22

or -ch2-ch-ch2.

I I oh oh r17 is hydrogen, lower alkyl, cycloalkyl or phenyl.

ris is hydrogen, lower alkyl, lower alkoxy or phenyl, or r17 and rls combined are: - (ch2) 2 -, - (ch2) 3 -, —ch = ch, or

R21 and R22 are independently selected from hydrogen and lower alkyl.

R23 is lower alkyl.

The present invention relates, in its broadest aspects, to substituted peptide compounds having the precipitated formula (I) and pharmaceutical compositions containing them.

The term "lower alkyl" used in the definition of various symbols refers to straight or branched chain radicals having up to 7 carbons. The preferred lower alkyl groups are those containing up to 4 carbons, methyl and ethyl groups being the most preferably used. Likewise, the terms "lower alkoxy" and "lower alkylthio" refer to lower alkyl groups attached to oxygen or sulfur.

The term "cycloalkyl" refers to saturated nuclei, from 3 to 7 carbon atoms, cyclopentyl and cyclohexyl being the most preferably used.

The term "halogen" refers to chloro, bromo and fluoro groups.

c = 0

30

where R40 and R6 (in the definition of X) are inhibiting groups, R40 being for example a benzyloxycarbonyl, and R <5 a benzyl. Removal of the inhibiting groups R40 and R6, for example by hydrogenation, provides the products of formula (I) where R6 is a hydrogen gene.

The carboxymethyl peptide ester of formula (II) is prepared by reacting the peptide ester of formula:

Rx O

I II H2N-CH-C-X (l)

(IV)

45

with tert.-butyl bromoacetate, then by introducing the group inhibiting R40, for example, by treatment with benzyl chloroformate.

The compounds of formula (I) can also be prepared by reacting a ketone of formula:

O O

He II

50 R2 - C — NH — CH — C — CH2-halo (V)

r3

where “halo” denotes Cl or Br, with an ester peptide of formula:

55 r rj o

I I II

HN-CH-C-X (VI)

(l)

60 in the presence of a base such as sodium bicarbonate, this reaction being followed by the elimination of the ester group R6.

The intermediate ketone of formula (V) can be prepared by treating a ketone of formula:

O

II

R40 - NH - CH - C - CH2-halo R3

(VII)

7

659,475

where R40 is an inhibiting group, such as benzyloxycarbonyl, with hydrobromic acid and acetic acid, then reacting with an acid halide of formula:

R, O halo-CH - C - O-Prot

(l)

(xvi)

O

II

R2 — C-halo

(VIII)

s where “Prot” is an easily removable ester inhibiting group, such as t-butyl, to give the ester:

in the presence of a base such as sodium bicarbonate.

The compounds of formula (I) can also be prepared by reacting an aminoketone of formula:

O

O

II

r

R2-C-NH-CH-C-CH,

I

r3

î1

o

-NH-CH-C-O-Prot (XVII) (l)

O OR

II II I

R2-C-NH-CH-C-CH2-NH

I

r3

(IX)

Removal of the ester inhibiting group provides the reaction product of formula XI.

In the above reactions, if any or all of the groups R, R1? R3 and R5 are as follows:

in particular the hydrochloride of this compound, with the haloketylamino or imino acid ester of formula:

Ri O

I II

halo-CH-C-X (L)

- (ch2 0h '- (cvr ^ -

(X)

where R6, in the definition of X, is an easily eliminable inhibiting ester group, and halo denotes Cl or Br.

The compounds of formula (I) can also be prepared by linking an aminoketone-carboxylic acid of formula:

(CH2) f "<0> - 0H '

Oh

- (ch2) -nh2, ~ r ~] [f jj '- (ch;) t —sh,

o o

r rj o n

I

h

- (CH2) r — OH, or - (CH2) r —NH —C,

r2-c-nh-ch-c-ch, -n-ch-c-oh

R *

(l)

(XI)

NOT,

NH

with the amino or imino acid ester of formula:

HX

in the presence of a binding agent, such as dicyclohexylcarbodi-imide where R0, in the definition of X, is an easily eliminable inhibiting group. Elimination of the inhibiting group R6 makes it possible to obtain the products of formula (I) where R6 is hydrogen.

The aminoketone of formula (IX) can be prepared by transforming the carboxyalkylamine of formula:

hoc-ch2

II

o

R

I

-N-R4

the hydroxyl, amino, imidazolyl, mercaptan or guanidi-nyl function should be inhibited during the reaction. Suitable inhibiting groups include the following: benzyloxycarbonyl, t-butoxy- (XII) carbonyl, benzyl, benzhydryl, trityl, etc., and nitro, in the case of guanidinyl. The inhibiting group is removed by hydrogenation, acid treatment, or other known methods, after completion of the reaction.

The ester products of formula (I), where R6 is lower alkyl, benzyl or benzhydryl, can be treated chemically, for example with sodium hydroxide in aqueous dioxane, or with trimethylsilyl bromide, to give the products of formula (I), where R6 is hydrogen. The benzyl and benzhydryl esters 45 can also be hydrogenated, for example by treatment with (XIII) of hydrogen, in the presence of a palladium catalyst on a carbon support.

The ester products of formula (I), where R6 is:

where R40 is an inhibiting group, such as benzyloxycarbonyl, in its acid chloride, then reacting with an oxazolone of formula (III) to give:

O

II

r, -c-nh-

O

r

-ch-I

R,

c-ch2-n-r4

(XIV)

Removal of the R40 inhibiting group, for example by hydrogenation provides the reaction product of formula (IX).

The amino ketone of formula (IX) where R is other than hydrogen can also be prepared by reacting the ketone of formula (V) with a substituted amine of formula:

R-NH,

(XV)

50 o

I!

-CH-0-C-R18

R1 7

55 can be obtained by using the peptide of formula (IV) or (VI) or the haloacetyl amino or imino acid ester of formula (X) in the abovementioned reactions, with such an ester group already in place. Such ester reaction products can be prepared by treating the peptide of formula (IV) or (VI) or the haloacetyl amino or iminoacid 60 ester of formula (X), where R6 is hydrogen, with an acid chloride such as :

O O

He II

-CH2-0-C-Cl or (H3Q3-C-O-C-CI

The aminoketone carboxylic acid of formula (XI) can be prepared by reacting the aminoketone of formula IX with a haloacetic acid ester of formula:

so as to inhibit the nitrogen atom. The inhibited compound is then caused to react, in the presence of a base, with a compound of formula:

659475

8

O where “Prot” is an inhibiting group such as

L-CH-0-C-R18 (XVIII) O

he 11

R "-C-0- (CH3) 3.

5

where L is a residue such as chlorine, bromine, tolylsulfonyl, etc., Preferably, this reaction is carried out in the presence of an agent after which the inhibiting group N is eliminated, by treatment with a bond, such as dicyclohexylcarbodiimide. Elimination of the acid group or hydrogenation. inhibiting N, for example by treatment with trifluoroacetic acid,

The ester products of formula (I), where R6 is makes it possible to obtain the peptide esters of formulas (IV) and (VI).

The haloacetyl amino or imino acid ester of formula (X) can be prepared by reacting the amino or imino acid ester of formula

(XII) with a haloacetyl halide of formula:

-CH-0-C-R18 R17

O

15 halo-CH2-C-halo (XXIII)

can also be obtained by treating the product of formula (I),

where R6 is hydrogen, by a molar excess of the compound of The products of formula (I), where R7 is an amino, can be formula (XVIII). obtained by reduction of the corresponding products of formula (I),

The ester products of formula (I), where R6 is where R7 is an azido.

The preferred compounds of the invention, as regards the peptide R2i O part of the structure of formula (I), are those in which:

R is hydrogen, lower straight or branched chain alkyl of 1 to 4 carbons, or phenyl. R22 R! is hydrogen, lower straight or branched chain alkyl

25 from 1 to 4 carbons, -CF3, - (CH2) r-NH2, where r is a number can be prepared by treating the product of formula (I), where R6 is an integer from 1 to 4,

hydrogen, with a molar excess of the compound of formula:

o "" 2 \ w —2 v-v ~ "2 v-v ° h '

-C C-O-R,

The ester products of formula (I), where R6 is - CH— (CH — OH) 2 or 35

"chir © '-ch2" @ - ° h,

I II 30

-C C-0-R23 (XIX) oh r22 -chj-n — rvsì, -ch ^ -7; n, -ch2-sh,

H H

-ch2-ch-ch2,

i .nh ni oh oh jp

- (ch2) 2 — s — ch3, - (ch2) 3nhc can be prepared by coupling the product of formula (I), where R6 40 is hydrogen, with a molar excess of the compound of formula:

CH2— (CH — O Prot) 2 (XX)

OH

or formula:

H2

O O

He II

-CH2-OH, -CH2-C-NH2, or - (CH2) 2-C-NH2

45

provided that ri is hydrogen, only if r is other than hydrogen.

CH2 — CH CH2 (XXI) R4 is hydrogen, cyclohexyl or phenyl.

III R5 is hydrogen, lower straight or branched chain alkyl

OH OProt OProt reliable from 1 to 4 carbons,

in the presence of a binding agent, such as dicyclohexylcarbodiimide, and then removing the hydroxyl inhibiting groups.

Esters of formula (I), where R6 is lower alkyl, may oh

"chf @, oh, -ch £ - (g ^ - oh,

Oh

be obtained from compounds of the carboxylic acid, that is to say 55 CH — N - (ch) nh in which r6 is hydrogen, by methods suitable for 2 ii [C-) J '2 l | | i '2'

esterification, for example, by treatment with halo N N

alkyl genide of formula rfl-halo, or an alcohol of formula I |

r6-oh. H h

The peptide esters of formulas (IV) and (VI) can be 60 —ch2 —sh, - (ch2) 2 —s — ch3,

obtained by coupling the hydrochloride of the amino or iminoacid ester of formula (XII), where r6 is for example benzyl, with the aminoh oo inhibited nacid n, of formula: - (ch2) 3nhc ^, -ch2- c-nh2 or- (ch2) 2-c-nh2.

Prot-N — CH — COOH (XXII) R "is hydrogen, lower straight or branched chain alkyl

(L) made from 1 to 4 carbons, an alkali metal salt,

9

659,475

0 O

II II

-CH-0 ~ C-R18, -CH2-C-0R23, -CH- (CH-OH) 2, or r17

-CH-CH-CH.

I I

OH OH

R23 is a straight or branched chain lower alkyl of 1 to 4 carbons, especially - C (CH3) 3.

R17 is hydrogen, straight or branched chain lower alkyl, or cyclohexyl.

Rls is a straight or branched chain lower alkyl of 1 to 4 carbons, or phenyl.

R is a lower straight or branched chain alkyl of 1 to 4 carbons, in particular - C (CH3) 3.

R, is hydrogen.

R7 is hydroxy.

R, is a straight or branched chain lower alkyl of 1 to 4 carbons, or a cyclohexyl.

R7 is an amino.

R7 is lower alkyl-O, where the lower alkyl is a straight or branched chain of 1 to 4 carbons.

R7 is - (CH,) 2 m

.- © V

13

where m is zero, one or two, and R13 is hydrogen, methyl, methoxy, methylthio, chloro, bromo, fluoro, or hydroxy.

R7 is -0- (ch2)

13

1-naphthyloxy or 2-naphthyloxy, where m is zero, one or two, and R13 is hydrogen, methyl, methoxy, methylthio, chloro, bromo, fluoro or hydroxy.

R7 is lower-S-alkyl, where lower alkyl is a straight or branched chain of 1 to 4 carbons.

R, is -S- (CH2); ^ g ^

13

1-naphthylthio or 2-naphthylthio, where m is zero, one or two, and R, 3 is hydrogen, methyl, methoxy, methylthio, chloro, bromo, fluoro or hydroxy.

Rs is a lower alkyl-O where the lower alkyl is a straight or branched chain of 1 to 4 carbons.

R8 is -0- (CH2)

13

where m is zero, one or two, and R13 is hydrogen, methyl, methoxy, methylthio, chloro, bromo, fluoro or hydroxy.

R8 is lower-S-alkyl, where lower alkyl is a straight or branched chain of 1 to 4 carbons.

Rs is

13

where m is zero, one or two, and RI3 is hydrogen, methyl, methoxy, methylthio, chloro, bromo, fluoro or hydroxy. R9 is phenyl, 2-hydroxyphenyl or 4-hydroxyphenyl. R10 represents two fluoro or chloro.

R10 represents two - Y - R16, where Y is O or S, R16 is a straight or branched chain lower alkyl of 1 to 4 carbons or the R16 groups joined to complete an unsubstituted 5 or 6 membered nucleus, or said nucleus wherein one or more of the available carbons have a methyl or dimethyl substituent.

Ru, R'n, R12 and R'12 are all hydrogen, or Rn is phenyl, 2-hydroxyphenyl, or 4-hydroxyphenyl, and R'n, RI2 and R'12 are hydrogen.

The preferred compounds of the invention, as regards the peptide part of the structure of formula (I), are those in which:

X is

-N-CH2-COOR6 R.

(CH,).

H, C

2 |

-NOT-

X

CH,

I 2

■ C-COOR.

(L)

h

/"NOT

O

-NOT

| (L)

COOR,

R is hydrogen or methyl.

Ri is hydrogen, methyl or - (CH2) 4NH2, in particular methyl, provided that R! or hydrogen, only if R is other than hydrogen.

R6 is hydrogen, straight or branched chain lower alkyl of 1 to 4 carbons, or an alkali metal salt.

R4 is cyclohexyl or phenyl.

R7 is hydrogen, cyclohexyl, lower alkoxy of 1 to 4 carbons,

40 -O- (CH2)

r ©, • o!

13

-S- (CVm

ï "© ..

13

'13

where m is zero, one or two, and RI3 is hydrogen, methyl, methoxy, methylthio. Cl, Br, F or hydroxy, the particularly preferred compounds being those in which R- is hydrogen.

t is two or three, preferably two.

The preferred compounds of the invention, with regard to the ketone part of the structure of formula (I), are those in which:

R2 is - (ch.)

2 IU

where m is zero, one or two and R14 is hydrogen, methyl, methoxy, methylthio, Cl, Br, F or hydroxy, especially phenyl. R3 is a straight or branched chain lower alkyl of 1 to 4 carbons, - (CH,) r —NH :,

• (CH2'm

R14

2 m rfo)

where m is zero, one or two, R, 4 is hydrogen, methyl, methoxy, methylthio, Cl, Br, F or hydroxy, and r is an integer from 1 to 4, especially benzyl.

The compounds of formula (I), where R6 is hydrogen, form salts with a variety of inorganic or organic bases. Pharmaceutically acceptable non-toxic salts are preferred, although other salts are also used in the isolation or

659475

10

product purification. These pharmaceutically acceptable salts include metal salts, such as sodium, potassium or lithium salts, alkaline earth metal salts, for example calcium or magnesium, and salts derived from amino acids such as arginine , lysine, etc. The salts are obtained by reacting the acid form of the compound with an equivalent of the base providing the desired ion in a medium in which the salt precipitates, or in an aqueous medium, then by lyophilizing.

Likewise, the compounds of formula (I), especially those in which R6 is an ester group, form salts with a variety of inorganic and organic acids. Here again, the non-toxic pharmaceutically acceptable salts are preferred, although other salts are also used in the isolation or purification of the product. These pharmaceutically acceptable salts include those formed with hydrochloric acid, methanesulfonic acid, sulfuric acid, maleic acid, etc. The salts are obtained by reacting the product with an equivalent amount of the acid in a medium in which the salt precipitates.

As indicated above, the peptide part of the molecule of the products of formula (I), represented by:

R Rj O

I I II -N-CH-C-X (L)

is of configuration L. An asymmetric center is also present in the ketone part of the molecule, when R3 is other than hydrogen. The compounds of formula (I) can therefore exist in diastereoisomeric forms or in mixtures thereof. The previously described processes can use racemic, enantiomeric or diastereomeric products as starting materials. When diastereomeric products are prepared, they can be separated by conventional methods of chromatography or fractional crystallization.

The products of formula (I) in which the imino acid nucleus is monosubstituted give rise to a cis-trans isomerism. The configuration of the final product will depend on the configuration of the substituents R7, Rs and R9 in the starting product of formula (XII).

The compounds of formula (I), and their pharmaceutically acceptable salts, are hypotensive agents. They inhibit the transformation of the decapeptide angiotensin I into angiotensin II, and are therefore used in the reduction or disappearance of hypertension linked to angiotensin. The action of the enzyme renin on an angio-tensinogen, a pseudoglobulin in the blood, produces angiotensin I. Angiotensin I is transformed, by the enzyme converting angiotensin (ACE), into angiotensin II . The latter is an active pressure substance which has been implicated as a causative agent in several forms of hypertension in various mammalian species, for example in humans. The compounds of the invention are involved in the sequence: angiotensinogen - "(renin) -" - angio-tension I-> angiotensin II, by inhibition of the angiotensin converting enzyme, and reduction or elimination of the formation of angiotensin II, an active pressure substance. Thus, by administration of a composition containing one (or a combination) of the compounds of the invention, angiotensin-dependent hypertension is relieved in species of mammals (for example humans) suffering from 'hypertension.

A single dose or, preferably, two to four doses distributed daily, scheduled on the basis of about 0.1 to 100 mg, preferably about 1 to 50 mg, per kg bodyweight per day, are suitable for reduce blood pressure. The substance is preferably administered orally, but can also be administered parenterally, for example subcutaneously, intramuscularly, intravenously or intraperitoneally.

The compounds of the invention can also be formulated in combination with a diuretic for the treatment of hypertension. A combined product comprising a compound of the invention and a diuretic can be administered in an effective amount, with a total daily dose of about 30 to 600 mg, preferably about 30 to 330 mg of a compound of the invention, and from about 15 to 300 mg, preferably about 15 to 200 mg of the diuretic, to a mammalian species in need of such treatment. As examples of diuretics envisaged for use in combination with a compound of the invention, mention is made of thiazide diuretics, for example chlorothiazide, hydrochlorothiazide, flumethiazide, hy-drofiumethiazide, bendroflumethiazide, methyclothiazide, trichloro-methiazide, polythiazide or benzthiazide, as well as ethacrynic acid and the compounds: ticrynafen, chlorthalidone, furosemide, muso-limine, bumetanide, triamterene, amiloride and Spironolactone, as well as salts of these compounds.

The compounds of formula (I) can be prepared, for use for reducing blood pressure, in compositions presented for example in the form of tablets, capsules or elixirs for oral administration, or in sterile solutions. or suspensions for parenteral administration. About 10 to 500 mg of a compound of formula (I) are mixed with carriers, carriers, excipients, binders, antiseptics, stabilizers, flavoring agents, all these products being physiologically acceptable, in the form of unit dose required in the standard pharmaceutical practice. The amount of active substance in these compositions or preparations must make it possible to obtain an appropriate dosage within the range of values specified.

The compounds of formula (I), where X represents

-NH-CH-C00R6,

r5

also have enkephalinase inhibiting activity and are used as analgesic agents. Thus, by administration of a composition containing one of the compounds of formula (I) or a combination thereof, or a pharmaceutically acceptable salt of these compounds, the pain is relieved in the mammalian host. A single dose or, preferably, two to four doses distributed daily, scheduled on the basis of about 0.1 to about 100 mg per kilogram of bodyweight per day, preferably about 1 to about 50 mg per kilogram per day, produce the desired analgesic activity. The composition is preferably administered orally, but can also be administered parenterally, for example subcutaneously.

The invention will now be described with reference to the examples below. Temperatures are given in degrees centigrade.

"LH-20" refers to a "Sephadex" chromatography gel, commercially distributed by "Pharmacia Fine Chemicals".

Example 1:

l- [N- [3- (benzoy lamino) -2-oxo-4-phenylbutyl] -L-alanyl] -L-proline, monohydrochloride a) L-alanyl-L-proline, phenylmethyl ester, acid salt p-toluenesul-fonique

N - [(1,1-dimethylethoxy) carbonyl] -L-alanine hydrochloride (310.7 g), L-proline, phenylmethyl ester (396.2 g), dicyclohexylcarbodiimide (338.6 g), l hydroxybenzotriazole hydrate (251.5 g), diisopropylethylamine (285.7 ml) and tetrahydrofuran (5 liters) are combined at 0 ° C (dicyclohexylcarbodiimide is added last) and stirred at room temperature for a night. The reaction mixture is filtered and concentrated. The residue is dissolved in 41 ethyl acetate and washed with 5% sodium bicarbonate (2x21), 5% potassium bisulfate (2 x 21) and water. The ethyl acetate layer is dried (MgS04) and concentrated. The residue is dissolved in 3 l of diethyl ether and left at 0 ° C overnight. The mixture is filtered and the filtrate is concentrated to give 620 g of N - [(1,1-dimethylethoxy) carbonyl] -L-alanyl-L-proline, phenylmethyl ester, in the form of a crude product. The N - [(1,1-dimethylethoxy) carbonyl] -L-alanyl-L-proline, phenylmethyl ester (275 g) is cooled in an ice bath and treated with 500 ml

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60

65

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cold trifluoroacetic acid (freshly distilled) in an argon atmosphere and stirred at room temperature for one hour. The mixture is concentrated in vacuo and made azeotropic twice with toluene. The crude salt of trifluoroacetic acid, which is in the form of an oil, is dissolved in 500 ml of ether and treated with gentle stirring with a solution of p-toluene sulfonic acid hydrate (1 equivalent ) dissolved in 6 l of ether. The resulting precipitate is collected by filtration. The solid product is dissolved in 1 l of methanol and treated with 4 l of ether and cooled. The resulting precipitate is combined and dried, which gives 300 g of L-alanyl-L-proline, phenyl methyl ester, p-toluenesulfonic acid salt; m.p. 156-158 ° C.

b) l- [N-f2- (1,1-dimethylethyl) -2-oxoethyl] -N - [(phenylmethoxy) -ccirbonyl] -L-alanyl] -L-proline, phenylmethyl ester

A mixture of L-alanyl-L-proline, phenylmethyl ester, p-toluenesulfonic (acid salt) (32.16 g, 72 mmol), tert.-butyl bro-moacetate (14.2 g, 72 mmol), triethylamine (20.1 ml, 144 mmol) and tetrahydrofuran (290 ml) is stirred at room temperature for 72 hours. Benzyl chloroformate (12.4 ml, 87.8 mmol) and triethylamine (12.5 ml, 87.8 mmol) are added and the mixture is stirred overnight. The reaction product is evaporated and partitioned between water and ethyl acetate. The products soluble in ethyl acetate are washed with dilute hydrochloric acid and then with water. The ethyl acetate solution is then evaporated and chromatographed on silica gel (2301 to 400 mesh - 0.063 to 0.037 mm) using the ethyl acetate / solvent system

hexane (1: 1) (pressure 0.07 bar) to give 19.5 g of l- [N- [2- (1,1-dimethylethylethoxy) -2-oxoethyl] -N - [(phenylmethoxy) carbo- nyl] -L-alanyl] -L-proline, phenylmethyl ester.

c) I- [N- (carboxymethyl) -N - [(phenylmethoxy) carbonyl] -L-alanyl] -L-proline, phenylmethyl ester

II [N- [2- (1,1-dimethylethoxy) -2-oxoethyl] -N - [(phenylmethoxy) carbonyl] -L-alanyl] -L-proline, phenylmethyl ester (18 g, 34.3) mmol) in trifluoroacetic acid (50 ml) and allowed to stand at room temperature for 1.5 hours. The mixture is evaporated and filtered on a small column of silica gel (200 g) using a mixture of chloroform / methanol / acetic acid solvent (9.6: 0.2: 0.2), which gives 11.4 g of 1- [N- (carboxymethyl) -N - [(phenylmethoxy) carbonyl] -L-alanyl] -L-proline, phenylmethyl ester.

d) I- [N- [3- (benzoy'lamino) -2-oxo-4-phenylbutyl] -N - [(phenylmethoxy) carbonyl] -L-alanyl] -L-proline, phenylmethyl ester

1 - [N- (carboxymethyl) -N - [(phenylmethoxy) carbonyl] -L-alanyl] -L-proline, phenylmethyl ester (7.0 g, 15.0 mmol) is dissolved in anhydrous tetrahydrofuran (50 ml ) and cooled in an ice bath, after which oxalyl chloride (1.57 ml, 18 mmol) and 4 drops of dimethylformamide are added. After 15 minutes, the reaction mixture is stirred at room temperature for an additional 1 hour. The mixture is evaporated, redissolved in tetrahydrofuran (30 ml) and the solution is cooled in an ice bath. This solution is poured dropwise, over a period of 5 minutes, into an ice-cold solution of 2-phenyl-4- (phenylmethyl) -5 (4H) -oxazolone (3.96 g, 15.75 mmol ) in tetrahydrofuran (24 ml). Triethylamine (2.5 ml, 17.1 mmol) is added and the reaction mixture is stirred at room temperature overnight. The mixture is filtered to separate the triethylamine hydrochloride, and the filtered solution of tetrahydrofuran is evaporated and redissolved in pyridine (16 ml). 4-Dimethylamino pyridine (50 mg) is added and the solution is stirred at room temperature for 3 hours. Crystallized acetic acid (16 ml) is added and the reaction mixture is heated at 100 ° C for 45 minutes. The reaction mixture is then cooled, evaporated in vacuo, dissolved in ethyl acetate and extracted with aqueous sodium bicarbonate and dilute hydrochloric acid. The ethyl acetate extract is chromatographed, after evaporation, on silica gel (230-400 mesh - 0.063 to 0.037 mm) using the ethyl acetate / benzene solvent system (4: 6) so as to obtain 4.9 g of 1- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N - [(phenyl-methoxy) carbonyl] -L-alanyl] -L-proline, phenylmethyl ester.

Analysis for C40H + 1N3O7 • 0.42H20:

Calculated: C 70.31 N 6.15 H 6.17 Found: C 70.31 N 6.13 H 6.08

e) l- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -L-alanyl] -L-proline, monohydrochloride

Dissolve in absolute ethanol (30 ml) and IN hydrochloric acid (2.25 ml) of 1- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N - [(phenylmethoxy ) carbonyl] -L-alanyl] -L-proline, phenylmethyl ester (1.0 g). A palladium-carbon catalyst (10%, 200 mg) is added and the solution is stirred under a hydrogen atmosphere overnight. The catalyst is separated by filtration and the solution is evaporated.

The crude product (700 mg) is combined with 300 mg of a crude product from a previous reaction on a reduced scale and passed through an LH-20 column (25.4 mm × 381 mm) in methanol. The fractions containing the desired product are combined, evaporated, dissolved in water and filtered. The clear aqueous solution is lyophilized, which gives 800 mg of ll [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -L-alanyl] -L-proline, monohydrochloride; [a] o = - 59.6 (c = 1.4, methanol); m.p. 98-130 ° C (with decomposition). Tic (silica gel, n-butanol / acetic acid / water, 4: 1: 1). Rr = 0.44.

Analysis for C25H30N3O5Cl • 0.75H20:

Calculated: C 59.88 H 6.33 N 8.38 Cl 7.07

Found: C 59.82 H 6.30 N 8.42 Cl 6.85

Example 2:

l- [N- [7-amino-3- (benzoylamino) -2-oxoheptvl] -L-alanyl] -L-proline, dihydrochloride a) N'-benzoyl-N6-] (phenylmethoxy) carbonyl] -L-lysine

To an ice-cold solution of N6 - [(phenylmethoxy) -carbonyl] -L-lysine (10.09 g, 36 mmol), in aqueous sodium hydroxide (IN, 26 ml), benzoyl chloride (5 ml, 43.2 mmol) and aqueous sodium hydroxide (4N, 10.8 ml) simultaneously in 5 portions, over a period of 30 minutes. The ice bath is removed and stirring is continued for a further 1.5 hours at room temperature. The reaction mixture is then extracted with ethyl acetate (rejected), the aqueous mother liquor is acidified with dilute hydrochloric acid and extracted with ethyl acetate. The ethyl acetate extract is concentrated and the residue is crystallized from ethyl acetate-hexane to give 12.9 g of N2-benzoyl-Nfi - [(phenylmethoxy) carbonyl] -L-lysine, m.p. 110-112; C (109 ° C).

b) 2-phenyl-4- [4 - [[(phenylmethoxy) carbonyl] amino] -butyl] -5 (4H) -oxazolone

The N2-benzoyl-N6 - [(phenylmethoxy) carbonyl] -L-lysine (11.53 mg, 30 mmol) is dissolved in tetrahydrofuran (55 ml) and the solution is stirred in an ice bath. To this reaction mixture, a solution of dicychlohexylcarbodiimide (6.8 g, 33 mmol) in tetrahydrofuran is added dropwise over a period of 15 minutes. The ice bath is removed after 1 hour and stirring is continued at room temperature for a further 18 hours. The di-cychlohexylurea is separated by filtration and the tetrahydrofuran is concentrated in vacuo. The residue is crystallized from ethyl acetate-hexane, which gives 9.4 g of 2-phenyl-4- [4 - [[(phenylmethoxy) carbo-nyl] amino] butyl] -5 (4H) - oxazolone; m.p. 72-73c C (68: C).

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c) l- [N- [3- (benzoylamino) -7 - [[(phenylmethoxy) carbonyl] amino] -2-oxoheptyl] -N- [(phenylmethoxy) carbonyl] -L-alanyl] -L-proline, phenylmethyl ester

The 1 - [N- (carboxymethyl) -N - [(phenylmethoxy) carbonyl] -L-alanyl] -L-proline, phenylmethyl ester (2.82 g, 6 mmol) from Example 1 (c) is dissolved in tetrahydrofuran (20 ml) and the solution is stirred in an ice bath. Oxalyl chloride (0.63 ml, 7.2 mmol) is added and then 4 drops of dimethylformamide. After stirring the reaction mixture in an ice bath for 20 minutes, this mixture is stirred at room temperature for an additional 1 hour. The solvents are removed in vacuo and the residue is redissolved in tetrahydrofuran (10 ml) and cooled in an ice bath. To this cold stirred solution is added a cold solution of 2-phenyl-4- [4 - [[(phenylmethoxy) carbo-nyI] amino] butyl] -5 (4H) -oxazoIone (2.2 g, 6 mmol ) in tetrahydrofuran (14 ml), then triethylamine (0.85 ml, 6 mmol). The ice bath is removed and the reaction mixture is stirred at room temperature overnight. The triethylamine hydrochloride is filtered off and the mother liquor is concentrated under vacuum. The latter is redissolved in pyridine (6 ml), 4-dimethylaminopyridine (30 mg) is added, then the solution is stirred at room temperature for 3 hours. Acetic acid (6 ml) is added and the reaction mixture is heated to 105 ° C for 45 minutes. This mixture is then evaporated, taken up in ethyl acetate and washed with water, dilute hydrochloric acid and aqueous sodium bicarbonate. After evaporation of the solvent, the crude product (3.8 g) is chromatographed (silica gel, 230 g) using ethyl acetate-hexane (4: 3), then ethyl acetate- hexane (2: 1) for elution to give 1.8 g of 1- [N- [3- (benzoylamino) -7 - [[(phenylmethoxy) carbonyl] amino] -2-oxoheptyl] -N- [(phenylmé-thoxy) carbonyl] -L-alanyl] -L-proline, phenylmethyl ester.

d) l- [N- [7-amino-3- (benzoylamino) -2-oxoheptyl] -L-alanyl] -L-proline, dihydrochloride

Dissolve in ethanol (75 ml) and aqueous hydrochloric acid (IN, 5 ml) of 1- [N- [3- (benzoylamino) -7 - [(phenylmethoxy) -carbonyl] amino] -2 -oxoheptyl] -N - [(phenylmethoxy) carbonyl] -L-alanyl-L-proline, phenylmethyl ester (1.3 g, 1.6 mmol). A palladium on carbon catalyst (10%, 450 mg) is added and the mixture is hydrogenated under atmospheric pressure overnight. The catalyst is filtered off and the solution is evaporated in vacuo. The residue is dissolved in water and lyophilized. The lyophilized product is triturated with ether to give 0.6 g of 1 - [N- [7-amino-3- (benzoylamino) -2-oxoheptyl] -L-alanyl] -L-proline hydrochloride; m.p. 80-155 ° C; [a] "= —50 ° (c = 1.1, methanol). Rr0.09 (silica gel, n-butanol / acetic acid / water, 4: 1: 1).

Analysis for C22H32N405 • 2HC1 • 1.5H20:

Calculated: C 49.63 H 7.00 N 10.52 Cl 13.55

Found: C 49.63 H 6.82 N 10.48 Cl 13.36

Example 3:

l- [N- [3- (benzoylamino) -2-oxoheptyl] -L-alanyl] -L-proline, monohydrochloride a) N-benzoyl-D, L-norleucine

D, L-norleucine (39.3 g, 300 mmol) is taken up in sodium hydroxide (2N, 150 ml) and, while stirring in an ice bath, sodium hydroxide (2N is added) , 150 ml) and benzoyl chloride (330 mmol, 38.3 ml) over a period of 30 minutes. The bath is removed and, after 1.5 hours, the reaction mixture is extracted with ether. The aqueous portion is acidified with 2N hydrochloric acid and the crystals are separated by filtration, which gives 68.9 g of N-benzoyl-D, L-norleucine; m.p. 131-133 ° C (125 ° C).

b) 4-butyl-2-phenyl-5 (4H) -oxazolone

The N-benzoyl-D, L-norleucine (40 g, 170 mmol) is taken up in tetrahydrofuran (300 ml) while stirring in an ice bath.

To this reaction mixture, a solution of dicyclohexylcarbodiimide (38.52 g, 187 mmol) in tetrahydrofuran (195 ml) is added dropwise over a period of 15 minutes. The ice bath is removed and stirring is continued at room temperature for an additional 18 hours. The dicyclohexylurea is separated by filtration and the tetrahydrofuran is concentrated in vacuo. The residue (31.7 g) is purified on silica in hexane: ether (2: 1) to give 32.1 g of 4-butyl-2-phenyl-5 (4H) -oxazolone.

0 c) l- [N- [3- (benzoylamino) -2-oxoheptyl] -N - [(phenylmethoxy) carbonyl] -L-alanyl] -L-proline, phenylmethyl ester

A solution of l- [N- (carboxymethyl) -N - [(phenylmethoxy) car-bonyl] -L-alanyl] -L-proline, phenylmethyl ester (1.41 g, 3 mmol) from Example 1 ( c) in tetrahydrofuran (10 ml) is again cooled in an ice bath. While stirring, oxalyl chloride (0.32 ml, 3.7 mmol) is added, then 4 drops of dimethylformamide. The reaction mixture is stirred in an ice bath for 20 minutes, then at room temperature for 1 hour. It is evaporated in vacuo, redissolved in tetrahydrofuran (5 ml) and cooled in an ice bath. A cold solution of 4-butyl-2-phenyl-5 (4H) -oxa-zolone (0.65 g, 3 mmol) in tetrahydrofuran (5 ml) is added dropwise, after which triethylamine is added ( 0.43 ml, 3.1 mmol). The reaction mixture is stirred at room temperature overnight. After the triethylamine hydrochloride has been filtered off, the tetrahydrofuran solution is concentrated in vacuo. The residue is redissolved in pyridine (3 ml), 4-dimethylamino pyridine (15 mg) is added and the reaction mixture is stirred at room temperature for 3 hours. Acetic acid (3 ml) is added and the reaction mixture is heated at 100 ° C for 40 minutes. This mixture is then evaporated, redissolved in ethyl acetate and washed with water, saturated sodium bicarbonate, dilute hydrochloric acid and water. After evaporation, the residue is chromatographed on silica gel, using the solvent system ethyl acetate: benzene (4: 6), which gives 0.8 g of 1- [N-35 [3- (benzoylamino) -2-oxoheptyl] -N - [(phenylmethoxy) carbonyl] -L-alanyl] -L-proline, phenylmethyl ester.

d) l- [N- [3- (benzoylamino) -2-oxoheptyl] -L-alanyl] -L-proline, mo-nochlorhydrate

40 Dissolve the l- [N- [3- (benzoylamino) -2-oxoheptyl] -N - [(phenyl-methoxy) carbonyl] -L-alanyl] -L-proline, phenyl methyl ester (0.96 g, 1 , 5 mmol) in ethanol (75 ml). Hydrochloric acid (IN, 2 ml) is added, then a palladium on carbon catalyst (10%, 200 mg). The solution is stirred under a hydrogen atmosphere overnight. The catalyst is filtered off, the ethanolic solution is evaporated and the residue is dissolved in water and lyophilized, which gives 0.62 g of 1- [N- [3- (benzoylamino) -2- oxoheptyl] -L-alanyl] -L-proline, monohydrochloride; m.p. 86-123 ° C; [a] o = 62.9 ° (c = 1.05, methanol). Rr0.57 (silica gel, n-butanol / acetic acid 50 / water, 4: 1: 1).

Analysis for C22H31N305 • HCl • 2H20:

Calculated: C 53.85 H 7.40 N 8.57 Cl 7.23

Found: C 53.85 H 7.20 N 8.73 Cl 7.29

55

Example 4:

l- [N- [3- (benzoy lamino) -2-oxo-4- (3-pyridinyl) butyl] -L-alanyl] -L-proline, dihydrochloride

60

a) 2- (Benzoylamino) -3- (3-pyridinyl) -2-propenoic acid

2-Phenyl-4- (3-pyridinylmethylen) -5 (4H) -oxazolone (3) is dissolved in acetic acid (24 ml) and in aqueous hydrochloric acid (0.5N, 150 ml) g, 12 mmol) (see Griffith et al., "J. Org. 65 Chem.", vol. 29, p. 2659). The reaction mixture is stirred overnight at room temperature. The mixture is then evaporated and re-evaporated with absolute ethanol. It is triturated with tetrahydrofuran, filtered, the filtered solid product then being re-crystallized by etha-

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noi absolute, which gives 2.8 g of _2- (benzoylamino) -3- (3-pyridi-nyl) -2-propenoic acid; m.p. 215-216 ° C (203 ° C).

b) 2- (Benzoylamino) -3- (3-pyridinyl) -2-propanoic acid

2- (benzoylamino) -3- (3-pyridinyl) -2-propenoic acid (14 g,

46 mmol) is dissolved in water (500 ml) and hydrogenated using a palladium on carbon catalyst (10%, 1.8 g) overnight. The catalyst is separated by filtration and the reaction mixture is evaporated to reduced volume (100 ml) and lyophilized to give 13.1 g of product. The lyophilized product is triturated with an absolute ethanol-ether mixture and filtered, which gives 12 g of 2- (benzoylamino) -3- (3-pyridinyl) -2-propanoic acid; m.p. 99-115 ° C.

c) 1- [N- [3- (benzoylamino) -2-0X0-4- (3-pyridinyl) -butyl] -N-

[(phenylmethoxy) carbonyl] -L-alanyl] -L-proline, phenylmethyl ester

The 1 - [N- (carboxymethyl) -N - [(phenylmethoxy) carbonyl] -L-alanyl] -L-proline, phenylmethyl ester (6.2 g, 13.1 mmol) from Example 1 (c) is dissolved in tetrahydrofuran (20 ml) and the solution is stirred in an ice bath. Oxalyl chloride is added, then 4 drops of dimethylformamide. After stirring this reaction mixture in an ice bath for 20 minutes, said mixture is further stirred at room temperature for an additional 1 hour. The solvents are removed in vacuo and the residue is redissolved in tetrahydrofuran (20 ml).

2- (benzoylamino) -3- (3-pyridmyl) -2-propanoic acid (4 g,

13 mmol) is suspended in tetrahydrofuran (45 ml) and while stirring in an ice bath, triethylamine (1.96 ml, 14 mmol) and dicyclohexylcarbodiimide (2.96 g, are added)

14 mmol). The reaction mixture is stirred at room temperature overnight, then filtered and the filtrate is evaporated to dryness. The residue obtained is dissolved in tetrahydrofuran (30 ml) and stirred in an ice bath. To this solution, the above-mentioned solution of 1- [N- (carboxymethyl) -N - [(phenylmethoxy) -carbonyl] -L-alanyl] -L-proline, phenylmethyl ester, acid chloride in tetrahydrofuran is added.

(20 ml). Triethylamine (1.9 ml, 13.6 mmol) is added and the reaction mixture is stirred at room temperature overnight, then filtered to separate the triethylamine hydrochloride. The filtrate is evaporated in vacuo, redissolved in pyridine (15 ml), 4-dimethylaminopyridine (65 mg) is added and the reaction mixture is stirred at room temperature for 3 hours. Acetic acid (16 ml) is added and the reaction mixture is heated at 100 ° C for 45 minutes. This mixture is then evaporated, redissolved in ethyl acetate and washed with aqueous sodium bicarbonate and with water. After evaporation, the ethyl acetate extract is chromatographed on silica gel using ethyl acetate for the elution, which gives 3.3 g of l- [N- [3- ( benzoylamino) -2-oxo-4- (3-pyri-dinyl) butyl] -N - [(phenylmethoxy) carbonyl] -L-alanyl] -L-proline, phenylmethyl ester.

d) l- [N- [3- (benzoylamino) -2-oxo-4- (3-pyridinyl) -butyl] -L-alanyl] -L-proline, dihydrochloride

The phenylmethyl ester product from part (c) (2.6 g, 3.84 mmol) is dissolved in ethanol (75 ml) and aqueous hydrochloric acid (IN, 8 ml) is added, then a palladium on carbon catalyst (10%, 0.6 g). After hydrogenation for 16 hours, a further 0.5 g of catalyst is added and the hydrogenation is continued for an additional 6 hours. The mixture is filtered, evaporated and combined with a similar reaction product obtained by hydrogenation of 0.8 g of the ester product from part (c). The hydrogenated product is then chromatographed on LH-20 in water, which provides the homogeneous product. An aqueous solution of this product is treated with aqueous hydrochloric acid (IN, 3 ml) and the solution is lyophilized to give 1.0 g of 1- [N- [3- (benzoylamino) -2-oxo-4 - (3-pyridinyl) butyl] -L-alanyl] -L-proline, dihydrochloride; m.p. 120-135 ° C; [a] o = —55.5 ° (c = 1.1, methanol). Rr0, ll (silica gel, n-butanol / acetic acid / water, 4: 1: 1).

Analysis for C24H28N405 • 2HC1 • 2H20:

Calculated: C 51.35 H 5.75 N 9.98 Cl 12.63

Found: C 51.35 H 5.84 N 9.96 Cl 12.84

Example 5:

l- [N- [3- (benzoylamino) -4- (4-hydroxyphenyl) -2-oxo-butyl] -L-alanyl] -L-proline, monohydrochloride a) 2-phenyl-4 - [[4- ( phenylmethoxy) phenyl] methyl] -5 (4H) -oxazo-lone

O-benzyl-L-tyrosine (11.0 g, 40.5 mmol) is taken up in 0.5N sodium hydroxide (81 ml) and water (80 ml) while shaking vigorously in an ice bath. To this are added, in 5 equal portions, a total of 52 ml of benzoyl chloride, 45 ml of IN sodium hydroxide and 400 ml of additional water over a period of 25 minutes. The bath is removed and the reaction is carried out for 2 hours at room temperature. The mixture is extracted twice with ethyl acetate. The aqueous portion is filtered, acidified with IN hydrochloric acid and the crystals are separated by filtration, which gives 12.9 g of N-benzoyl-O-benzyl-L-tyrosine: m.p. 166-168 "C (162 ° C).

This N-benzoyl-O-benzyl-L-tyrosine (12.76 g, 35 mmol) is taken up in anhydrous tetrahydrofuran (50 ml) while stirring in an ice bath. Dicyclohehylcarbodiimide (7.7 g 37.4 mmol) in tetrahydrofuran (18 ml) is added to this solution dropwise. After 20 minutes, the ice bath is removed and the reaction is allowed to proceed overnight at room temperature. The dicyclohexylurea is filtered off and the filtrate is concentrated to dryness. The crude product is crystallized from ether / hexane to give 10.26 g of 2-phenyl-4-14- (phenylmethoxy) phenyl] methyll-5 (4H) -oxazolone; m.p. 85-87 ° C (83 ° C).

b) l- [N- [3- (benzoylamino) -2-oxo-4- [4- (phenylmethoxy) -phenyl] bu-tyl] -N- [(phenylmethoxy) carbonyl] -L-alanyl] -L- proline, phenylmethyl ester

The 1 - [N- (carboxymethyl) -N - [(phenylmethoxy) carbonyl] -L-ala-nyl] -L-proline, phenylmethyl ester (2.82 g, 6 mmol) from example (c) is dissolved in tetrahydrofuran (20 ml) and the solution is stirred in an ice bath. Oxalyl chloride (0.63 ml, 7.2 mmol) is added and then 4 drops of dimethylformamide. After stirring this reaction mixture in an ice bath for 20 minutes, stirring is continued at room temperature for an additional 1 hour. The solvents are removed in vacuo and the residue is redissolved in tetrahydrofuran (10 ml) and cooled in an ice bath. To this cold stirred solution is added a cold solution of 2-phenyl-4 - [[4- (phenylmethoxy) -phenyl] methyl] -5 (4H) -oxazolone (2.14 g, 6 mmol) in tetrahydrofuran (40 ml). Triethylamine (0.84 ml, 6 mmol) is added and a basic atmosphere is maintained for the duration of the reaction by addition of the required required amount of triethylamine. The reaction mixture is stirred at room temperature overnight. It is then filtered and the filtrate is evaporated and redissolved in pyridine (7 ml). 4-Dimethylamino pyridine (30 mg) is added and the reaction mixture is stirred for 3 hours at room temperature. Acetic acid (7 ml) is added and the reaction mixture is heated at 100 ° C for 45 minutes. This mixture is then evaporated, the residue is redissolved in ethyl acetate and washed with saturated sodium bicarbonate and dilute hydrochloric acid. The neutral ethyl acetate extract is evaporated and chromatographed on silica gel (300 g) using the ethyl acetate: benzene solvent system (6.5: 3.5) to give 2.7 g of l- [N- [3- (benzoylamino) -2-oxo-4- [4- (phenylmethoxy) -phenyl] butyl] -N - [(phenylmethoxy) carbo-nyl] -L-alanyl] -L-proline, phenylmethyl ester.

c) 1- [N- [3- (benzoylamino) -4- (4-hydroxyphenyl J-2-oxobutyl] -L-alanyl] -L-proline, monohydrochloride

The ester product from part (b) (1.8 g, 2.26 mmol) is dissolved in ethanol (150 ml) containing hydrochloric acid.

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than aqueous (IN, 3.5 ml). A palladium catalyst is added to the inhibiting groups Rj in Examples 20, 36 to 39 and 41, the charcoals (10%, 500 mg) and the solution is stirred under an atmosphere of inhibiting groups R3 in Examples 43 and 44 and inhi- groups

of hydrogen overnight. This solution is then evaporated, bants R5 in examples 49, 50 and 52 to 55 are eliminated at the last

then dissolved in water and lyophilized, which gives the l- [N- [3- third step, in the synthesis. The R6 ester groups indicated in the

(benzoylamino) -4- (4-hydroxyphenyl) -2-oxobutyl] -L-alanyl] -L- Examples 57 to 62 are not eliminated.

proline, monohydrochloride; m.p. 118-152 ° C; ["] □ = —63.1 ° (c =

1.07, methanol). Rf 0.47 (silica gel, n-butanol / acetic acid / water,

4.1.1 j Example 63:

,, t, XI _ TT _ l- [N - [(S) -3- (benzoylamino) -2-oxo-4-phenylbutyl] -L-alanyl] -L-

Analysis for C25H29N3Ofi • HCl • H20: „monohydrochloride

Calculated: C 57.42 H 6.16 N 8.04 Cl 6.78 Found: C 57.42 H 6.03 N 8.04 Cl 7.11

a) (S) -3-amino-1-chloro-4-phenyl-2-butanone, hydrobromic acid

The (S) - [3-chloro-2-oxo-1- (phenylmethyl) propyl] -carbamic acid, phenylmethyl ester (51.4 g) is dissolved in a mixture of acid Examples 6-62 15 acetic ( 252 ml) and hydrobromic acid in acetic acid

Based on the method of Examples 1-3 and 5, the peptide <3> 45N '348 ml> and maintained at room temperature for the ester indicated in column I is treated to give the carboxym- 1'5 hour' The melan ®e rational is then concentrated in vacuo and thyl peptide ester indicated in column II. Transformation to Preclplte with Fether> ce ^ Fournlt 36'6 "of (S) -3-amino-1-chlo-son acid chloride followed by reaction with the oxazolone of ro-4-phenyl-2 -butanone, hydrobromic acid; m.p. 177-179 ° C

fl75 ° W

column III provides the N-inhibited ester product of column IV. The eli- 20 v> '

mination of the N-inhibiting group and the ester group provides the final product of column V where Re is hydrogen. k) (S) -N- [3-chloro-2-oxo-1- (phenylmethyl) propyl] benzamide

(S) -3-amino-1-chloro-4-phenyl-2-butanone, bromhydri-j acid (36.3 g, 130.3 mmol) is suspended in 520 ml of tetra-

Anhydrous hydrofuran and 18.2 ml of triethylamine (130.3 mmol) while Ri O with stirring for 10 minutes. The mixture is placed in an ice bath and 15.2 ml of benzoyfe chloride are added, then 10.95 g of ^ L) V sodium bicarbonate. After 5 minutes, the ice bath is removed, and the reaction mixture is maintained at room temperature for 1.5 hours. The reaction mixture is then concento /. II very vacuum, and the residue is taken up in 11 of aqueous methanol (at

10% water). The precipitate is combined, filtered and washed with methanol, which gives 25.3 g of (S) -N- [3-chloro-2-oxo-1- (phenylimethyl) propyl] -HO-Ç-CH2-N- CH-CX benzamide; m.p. (160 ° C) 170-172 ° C (decomposition); [c] g = -129 (L) 35 (c = 1.7, dimethylformamide).

H, N — CH — C — X

Ri O

O

C-0-CH2

II

O

<Ô>

c) 1- [N - [(S) -3- (benzoylamino) -2-oxo-4-phenylbutyl] -L-alanyl] -L-proline, 1,1-dimethylethyl ester

40 L-alanyl-L-proline, 1,1-dimethylethyl ester (2.42 g, 10 mmol), Col III sodium bicarbonate (840 mg) and (S) -N- [3-chloro-2 -oxo-l-

N (phenylmethyl) propyl] benzamide (3.01 g) are combined in 50 ml of dimethylformamide under an argon atmosphere, at room temperature am-R2-Ç Ç-R3 and with stirring overnight. The reaction mixture q C = O 45 is then concentrated under vacuum to approximately half of its initial volume and the residue is taken up in ethyl acetate and washed with saturated sodium bicarbonate to give 2.25 g of product. gross. This Col. IV product is taken up in ethyl acetate: methanol (95: 5) and applied

q R O qué on a column of silica gel (135 g) and eluted with ethyl acetate

II I 1 II so le: methanol (95: 5) to give 860 mg of l- [N - [(S) -3- (benzoylami-

R3 - ÇH — C — CH 2 — N — CH — C — X no) -2-oxo-4-phenylbutyl] -L-alanyl] -L-proline, 1,1-dimethylethyl

NH I (L) / _ \ ester.

co-(

c = o o

d) l- [N - [(S) -3- (benzoylamino) -2-oxo-4-phenylbutyl] -L-alanyl] -L-55 proline, monohydrochloride

L- [N - [(S) -3- (benzoylamino) -2-oxo-4-phenylbutyl] -L-alanyl] -

Col y L-proline, 1,1-dimethylethyl ester (740 mg, 1.46 mmol) is dissolved in a solution of hydrochloric acid in acetic acid (1.5N,

O Ri O 10 ml) and kept at room temperature for 30 minutes. The

_ £ j_j_ç_çjj __çjj_ç_x 60 solution is concentrated, taken up in water, filtered and lyophilized, which

I (L) gives 600 mg of 1- [N- (S) -3- (benzoylamino) -2-oxo-4-phenylbutyl] -

NH L-alanyl] -L-proline, monohydrochloride; m.p. 83-163 ° C; [a] "=

^ _0 —109 ° (c = 1.04, methanol). Rf 0.6 (silica gel, butanol / acetic acid

Tick / water, 4: 1: 1).

R2 65

Analysis for C25H2, N305 • HCl • 1.65H20:

Calculated: C 57.99 H 6.48 N8.12 Cl 6.85

(Table on following pages) Found: C 57.99 H 6.39 N 8.09 Cl 6.95

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34

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40

41

42

43

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58

59

60

61

62

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25

659,475

Example 64:

(S) -1- [N2- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -L-lysyl] -L-proline, dihydrochloride a) l- [N6 - [(1,1-dimethylethoxy) carbonyl] -N2- [(phenylmethoxy) car -bonylJ-L-lysyl] -L-proline, 1,1-dimethylethyl ester

N6 - [(1,1-dimethylethoxy) carbonyl] -N2 - [(phenylmethoxy) car-bonyl] -L-lysine (9.51 g) and hydroxybenzotriazole (3.825 g) are poured into 25 ml of dimethylformamide, while stirring in an ice bath, under an argon atmosphere. To this product is added L-proline, 1,1-dimethylethyl ester (4.49 g) then N, N'-diisopro-pylethylamine (2.2 ml) and dicyclohexylcarbodiimide (5.15 g). After 15 minutes, the bath is removed and the reaction is allowed to continue for 6.5 hours at room temperature. Dimethylformamide is removed in vacuo. The residue is taken up in ethyl acetate and the dicyclohexylurea is separated by filtration. The filtrate is washed neutral with 10% potassium bisulfate and saturated sodium bicarbonate. The crude product (13.26 g) is purified on a column of silica gel, eluting with ethyl acetate: hexane (2: 1), which gives 13.0 g of 1- [N6 - [( 1,1-dimethylethoxy) carbonyl] -N2 - [(phenyl-methoxy) carbonyl] -L-lysyl] -L-proline, 1,1-dimethylethyl ester.

b) (Sj-1- [N2- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N6 - [(1,1-dimethylethoxy) carbonyl] -L-lysyl] -L-proline, 1, 1-dimethylethyl ester

The ester product from part (a) is reduced in ethanol using a palladium on carbon catalyst (10%) to give 3.99 g of l- [N6- [l, l- ( dimethylethoxy) -carbonyl] -L-lysyl] -L-proline, 1,1-dimethylethyl ester. This product is introduced into 40 ml of dimethylformamide and treated with (S) -N- [3-chloro-2-oxo-1- (phenylmethyl) propyl] benzamide (3.01 g) from Example 63 (b ) and sodium bicarbonate (840 mg). After stirring for 18 hours at room temperature, the reaction mixture is concentrated under vacuum, taken up in ethyl acetate and washed with saturated sodium bicarbonate. The crude product (7.0 g) is purified on a silica gel column, eluting with ethyl acetate: 1% methanol to give 1.7 g of (S) -l- [N2- [3 - (benzoylamino) -2-oxo-4-phenylbutyl] -N6 - [(1,1-dimethylethoxy) carbonyl] -L-lysyl] -L-proline, 1,1-dimethylethyl ester.

c) (S) -1 - [N2- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -L-lysyl] -L-proline, dihydrochloride

The ester product from part (b) (1.6 g) is treated for 30 minutes at room temperature with 20 ml of 1.5N hydrochloric acid: acetic acid, concentrated to dryness and triturated into solid product with ether, so as to obtain 1.37 g of crude product. This product is taken up in water, filtered through a millipore filter and lyophilized to give 1.28 g of final product. Further purification is carried out on an LH20 column in water, which provides 740 mg of (S) -l- [N2- [3- (benzoylamino) -2-oxo-4-phenylbu-tyl] -L- lysyl] -L-proline, dihydrochloride; m.p. 120-180 ° C; [a] o = - 84.8 ° (c = 1.05, methanol). Rr0.61 (traces at 0.9) (silica gel, chloroform / methanol / acetic acid, 60.40: 38:20).

Analysis for C28H3 (; N4Os ■ 2HC1 • 3H20:

Calculated: C 52.98 H 6.98 N 8.83 Cl 11.17

Found: C 52.98 H 6.93 N 8.66 Cl 11.31

Example 65:

N- [N- [[(S) -3- (benzoylamino) -2-oxo-4-phenylbutylJ-L-alanylJ-N-cyclohexylglycine, monohydrochloride a) N-cyclohexylglycine, 1,1-dimethylethyl ester

Cyclohexylamine (70.35 ml) and sodium bicarbonate (12.9 g) are suspended, with stirring, in 200 ml of absolute ethanol, maintaining stirring in an ice bath. To this product, bromoacetic acid, 1,1-di-methylethyl ester (20.78 ml) is added dropwise. The ice bath is removed. After

24 hours at room temperature, the reaction mixture is concentrated to dryness, taken up in chloroform and washed with water. The crude product (42 g) is chromatographed on silica gel, eluting with ethyl acetate: hexane (2: 1) to give 27.4 g of N-cyclohexylglycine, 1,1-dimethylethyl ester.

b) N-cyclohexyl-N- [N- [(phenylmethoxy) carbonylJ-L-alanyl] glycine, 1,1-dimethylethyl ester

N - [(phenylmethoxy) carbonyl] -L-alanine (4.46 g), N-cyclohexylglycine, 1,1-dimethylethyl ester (4.26 g), hydroxybenzotriazole (3.06 g), dicyclohexylcarbodiimide (4.12 g) and triethylamine (2.8 ml) are stirred in 40 ml of dimethylformamide at room temperature for 20 hours. The reaction mixture is then concentrated under vacuum, taken up in ethyl acetate, the dicyclohexylurea is separated by filtration and the filtrate is washed to neutral with 10% potassium bisulfate and saturated sodium bicarbonate to give 5.9 g of crude product. Crystallization with ether: hexane provides 3.97 g of N-cyclohexyl-N- [N - [(phenylme-thoxy) carbonyl] -L-alanyl] glycine, 1,1-dimethylethyl ester; m.p. 104-105 ° C.

c) N- (L-alanyl) -N-cyclohexylglycine, 1,1-dimethylethyl ester

The ester product from part (b) (3.9 g) is taken up in methanol with a palladium on carbon catalyst (10%, 700 mg) and stirred under a hydrogen atmosphere for 4 hours. The reaction mixture is filtered and concentrated to dryness to give 2.65 g of N- (L-alanyl) -N-cyclohexylglycine, 1,1-dimethylethyl ester, in the form of a crude product.

d) N- [N - [(S) -3- (benzoylamino) -2-oxo-4-phenylbutyl] -L-alanylJ-N-cyclohexylglycine, 1,1-dimethylethyl ester

The crude ester product from part (c) (2.6 g), sodium bicarbonate (764 mg) and (S) -N- [3-chloro-2-oxo-l- (phenylme-thyl) propyl] benzamide (2.75 g) of Example 63 (b) are stirred for 20 hours in 25 ml of dimethylformamide. The reaction mixture is concentrated to dryness, taken up in ethyl acetate and washed with saturated sodium carbonate to give 4.7 g of crude product. By purification on a column of silica gel, with elution with ethyl acetate: methanol (99: 1), 1.5 g of N- [N - [(S) -3- (benzoylamino) -2 are obtained. -oxo-4-phenylbutyl] -L-alanyl] -N-cyclohexylgly-cine, 1,1-dimethylethyl ester.

e) N- [N- [(S) -3- (benzoylamino) -2-oxo-4-phenylbutyl] -L-alanylJ-N-cyclohexylglycine, monohydrochloride

The ester product of part (d) (500 mg) is treated for 30 minutes with 5 ml of 1.5N hydrochloric acid: acetic acid, then concentrated to dryness at room temperature. The crude product is taken up in methanol and purified on an LH20 column to give 413 mg of product. This is present in a semi-crystallized state in acetonitrile: ether to give N- [N - [(S) -3- (benzoylamino) -2-oxo-4-phenylbutyl] -L-alanyl] -N-cyclohexylglycine, monohydrochloride; m.p. 154-157 C (132 "C); [a] o = —67.4 (c = 1.35, methanol). Rr0.60 (minor impurities at 0.92) (silica gel, chloroform / me-thanol / concentrated ammonia, 30: 10: 2).

Analysis for C28H35N305 • HCl • H20:

Calculated: C 61.35 H 6.99 N 7.67 Cl 6.47

Found: C 61.08 H 6.79 N 7.65 Cl 6.46

Example 66:

N-f N - [(S) -3- (benzoylamino) -2-oxo-4-phenylbutylJ-L-alanyl] -N-phenylglycine, monohydrochloride a) N-phenylglycine, 1,1-dimethylethyl ester

A solution of triethylamine (11.25 g, 0.11 mol) and aniline (9.3 g, 0.10 mol) in ether (100 ml) under an argon atmosphere is cooled to 0J C in a ice bath. Bromoacetic acid, 1,1-dimethylethyl ester (18 g, 0.093 mol) is added to this solution on

5

10

15

20

25

30

35

40

45

50

55

60

65

659475

26

a period of 30 minutes. The resulting mixture is stirred at 0 ° C for 1 hour, then brought to room temperature and stirred overnight. The solution is filtered and rinsed with ether, and the filtrate is concentrated, giving 6.9 g of a yellow oil. Rr 0.6, 0.7 (silica gel, ethyl acetate). By chromatography on LPS-1, using hexane: ethyl acetate (7: 3) as eluent, 2.3 g of N-phenyl-glycine, 1,1-dimethyl ester, are obtained in the form of a pale yellow liquid. Rr 0.7 (silica gel, ethyl acetate).

b) N-phenyl-N- [N - [(phenylmethoxy) carbonyl] -L-alanyl] glycine, 1,1-dimethylethyl ester

A solution of N - [(phenylmethoxy) carbonyl] -L-alanine (2.8 g, 12.7 mmol) in anhydrous tetrahydrofuran (50 ml) under argon, is cooled in a dry ice-ethanol bath. To this solution are added N-methylmorpholine (1.28 g, 12.7 mmol) and isobutylchloroformate (1.73 g, 12.7 mmol). After 20 minutes, N-phenylglycine, 1,1-dimethyl ester (3.7 g, 12.7 mmol) is added. The resulting mixture was stirred at -20 ° C for 1 hour, then at room temperature overnight. The mixture is partitioned between ethyl acetate and IN hydrochloric acid. The organic layer is washed successively with IN hydrochloric acid and 10% sodium bicarbonate, dried (MgSO 4) and concentrated. By chromatography on LPS-1 with elution with a gradient of ethyl acetate: hexane (3: 1 1: 1), 4.0 g of N-phenyl-N- [N - [(phenylmethoxy) carbonyl] are obtained. -L-alanyl] glycine, 1,1-dimethylethyl ester, in the form of a clear oil. Rr 0.4 (silica gel, ethyl acetate).

c) N- (L-alanyl) -N-phenylglycine, 1,1-dimethylethyl ester

A solution of the ester product from part (b) (4.0 g, 9.7 mmol) in ethanol (125 ml) with 10% palladium on carbon catalyst is stirred under a stream of hydrogen for 18 hours. The solution is filtered, concentrated, dissolved in ethyl acetate and extracted with IN hydrochloric acid. The aqueous layer is treated with sodium bicarbonate until basic reaction, and extracted with ethyl acetate. The combined ethyl acetate extracts are dried (MgSO4) and concentrated to give 1.3 g of N- (L-alanyl) -N-phenylglycine, 1,1-dimethylethyl ester, as a white solid . Rf 0.52, minor spot at 0.64 (silica gel, ethyl acetate: methanol, 1: 1).

d) N- [N - [(S) -3- (benzoylamino) -2-oxo-4-phenylbutyl] -L-alanyl] -N-phenylglycine, 1,1-dimethylethyl ester

To a stirred solution of (S) -N- [3-chloro-2-oxo-l- (phenylme-thyl) propyl] benzamide (1.4 g, 4.7 mmol), of Example 63 (b) , and N- (L-alanyl) -N-phenylglycine, 1,1-dimethylethyl ester (1.3 g, 4.7 mmol) in anhydrous dimethylformamide, sodium bicarbonate (0.38 g, 4 , 7 mmol) and sodium iodide (0.7 g, 4.7 mmol). After stirring overnight at room temperature, the mixture is concentrated, dissolved in ethyl acetate and filtered. The filtrate is washed with 10% sodium bicarbonate, dried (MgSO4) and concentrated in the form of a yellow oil. By chromatography on LPS-1, eluting with a gradient of ethyl acetate: hexane (1: 1) to ethyl acetate, 1.8 g of N- [N - [(S) -3- are obtained. (benzoylamino) -2-oxo-4-phenylbutyl] -L-alanyl] -N-phenylglycine, 1,1-dimethylethyl ester. Rr0.34 (silica gel, ethyl acetate).

e) N- [N - [(S) -3- (benzoylamino) -2-oxo-4-phenylbutyl] -L-alanylJ-N-phenylglycine, hydrochloride

A solution of ester product from part (d) (1.6 g, 2.9 mmol) in hydrochloric acid / acetic acid (1.77N, 17.0 ml) is stirred at room temperature for 30 minutes. The solution is concentrated and the residue is triturated with ether to give a pale yellow solid product. By recrystallization from me-thanol / ether, 0.84 g of N- [N - [(S) -3- (benzoylamino) -2-oxo-4-phenylbutyl] -L-alanyl] -N-phenylglycine is obtained. , monohydrochloride in the form of a white crystalline solid, mp 147-159 ° C (decomposition). Rr 0.8 (silica gel; butanol / acetic acid / water, 1: 1: 1. [A] D = - 12.7 ° (c = 1.5, methanol).

Analysis for C28H2gN305 • HCl • 0.65H20:

Calculated: C 62.77 H 5.89 N 7.84 Cl 6.62 Found: C 62.77 H 5.70 N 7.84 Cl 6.12

Example 67:

(S) -l- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N-methyl-L-alanyl] -L-proline, monohydrochloride a) N - [(phenylmethoxy) carbonyl] - L-alanine

L-alanine (89.1 g, 1 mol) is dissolved in 2N sodium hydroxide (500 ml) and cooled to 0 ° C. To this solution is added dropwise simultaneously over a 1 hour period, benzylchloroformate (204.5 g, 1.2 mol) and 4N sodium hydroxide (250 ml). The reaction mixture is stirred overnight (from 0 ° C to room temperature) and washed with ethyl acetate (2 x 500 ml). The aqueous portion is acidified to pH 2.0 with 6N hydrochloric acid and extracted with ethyl acetate (3 x 600 ml). The combined ethyl acetate extracts are dried (Na2SO4), concentrated on a rotary evaporator, and the solid residue is triturated with petroleum ether to give 194.0 g of N - [(phenylmethoxy) carbo-nyl ] -L-alanine as a white solid.

b) N-methyl-N- [(phenylmethoxy) carbonyl] -L-alanine

To a cold solution (0 ° C) of N - [(phenylmethoxy) carbonyl] -L-alanine (22.3 g, 0.1 mol) and methyl iodide (50 ml, 0.8 mol) in tetrahydrofuran (250 ml), a solution of sodium hydride (14.25 g, 0.3 mol) is added carefully and with gentle stirring. The suspension is stirred overnight under a nitrogen atmosphere (from 0 ° C to room temperature), then it is poured slowly into saturated sodium bicarbonate (200 ml), and diluted with ethyl acetate (300 ml) and the layers are separated. The organic layer is again extracted with saturated sodium bicarbonate. The combined aqueous layers are acidified to pH 2.0 with 10% potassium bisulfate and extracted with ethyl acetate. The combined ethyl acetate extracts are dried (Na2SO4) and concentrated in vacuo to give a dark oily residue (23.0 g).

This crude acid is treated with dicyclohexylamine (20 ml) in ether (150 ml). The crude salt resulting from dicyclohexylamine is combined (37.0 g), recrystallized from chloroform / ether (35.0 g) and converted back to the acid by partitioning between IN hydrochloric acid / ethyl acetate, which gives a pale yellow oil which crystallizes after leaving it to stand (17.4 g).

By recrystallization (11.2 g) from ethyl acetate / petroleum ether, 4.0 g of N-methyl-N - [(phenylmethoxy) carbonyl] -L-alanine is obtained, in the form of a white crystallized product; m.p. 65-65.5 ° C; [a] "= —31.1 ° (c = 2, acetic acid).

c) l- [N-methyl-N - [(phenylmethoxy) carbonyl] -L-alanyl] -L-proline, 1,1-dimethylethyl ester

To a solution of N-methyl-N - [(phenylmethoxy) carbonyl] -L-alanine (4.74 g, 20 mmol) in distilled tetrahydrofuran (50 ml), L-proline, 1,1-dimethylethyl is added ester (3.42 g, 20 mmol), hydroxybenzotriazole hydrate (3.06 g, 20 mmol) and dicyclohexylcarbodiimide (4.12 g, 20 mmol). The reaction mixture is stirred overnight, the precipitated dicyclohexylurea is filtered off and the filtrate is concentrated. Dissolve the residue in ethyl acetate (50 ml) and wash with saturated sodium bicarbonate (twice), 10% potassium bisulfate (twice) and water (twice) , dried (Na2SO4) and concentrated, which gives 6.4 g of l- [N-methyl-N - [(phenylmethoxy) carbonyl] -L-alanyl] -L-proline, 1,1-dimethylethyl ester, in the form of an oily residue.

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d) l- (N-methyl-L-alanyl) -L-proline, 1,1-dimethylethyl ester

A mixture of ester product from part (c) (6.4 g, 16.4 mmol) and 0.8 g of 10% palladium catalyst on carbon in ethanol (95%, 150 ml) is hydrogenated to atmospheric pressure overnight. The catalyst is removed by filtration and the filtrate is evaporated. The oily residue formed solidifies by drying under high vacuum, into an oily solid residue (3.8 g). By trituration with ether, 1.5 g of l- (N-methyl-L-alanyl -) - L-proline, 1,1-dimethylethyl ester, monohydrochloride, are obtained in the form of a white solid; Rf 0.44 (silica gel, 20% methanol / chloroform). [a] "= -102.1 ° (c = 2, acetic acid).

The ethereal filtrate provides 2.3 g of 1- (N-methyl-L-alanyl) -L-proline, 1,1-dimethyl ester, as an oil; Rr 0.44 (silica gel, 20% methanol / chloroform). [a] "= —101.6 ° (c = 2, acetic acid).

e) (S) -1- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N-methyl-L-alanyl] -L-proline, 1,1-dimethylethyl ester

Stirred at room temperature overnight under a nitrogen atmosphere, a reaction mixture containing l- (N-methyl-L-alanyl) -L-proline, 1,1-dimethylethyl ester (2.1 g, 8 , 19 mmol), (S) -N- [3-chloro-2-oxo-1- (phenylmethyl) propyl] benzamide (2.46 g, 8.19 mmol) from Example 63 (b), a excess sodium bicarbonate, and sodium iodide (1.22 g, 8.10 mmol) in dimethylformamide (15 ml). The reaction mixture is concentrated, and the residue is partitioned between water / ethyl acetate; the layers are separated and the aqueous layer is extracted once again with ethyl acetate. The combined organic extracts are washed with saturated sodium bicarbonate and with water, dried (Na2SO4) and concentrated in an oily residue (3.5 g). By rapid chromatography (200 g of silica gel, 1% methanol / ethyl acetate), 2.8 g of (S) -l- [N- [3- (benzoylami-no) -2-oxo are obtained -4-phenylbutyl] -N-methyl-L-alanyl] -L-proline, 1,1-dimé-thyléthyl ester, in the form of a yellow oil.

f) (S) -1-fN- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N-methyl-L-alanyl] -L-proline, monohydrochloride

The ester product from part (e) (1.4 g, 2.7 mmol) is treated with 2N hydrochloric acid / acetic acid (20 ml). After stirring for 2 hours at room temperature, the reaction mixture is concentrated under reduced pressure, and the oily residue formed is triturated with ether (four times) to give 1.05 g of (S) -l- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N-methyl-L-alanyl] -L-proline, monohydrochloride, in the form of a whitish solid; m.p. 125-135 ° C; Rf 0.24 (silica gel; n-butanol / acetic acid / water, 4: 1: 1). [<x] d = -87 ° (c = 1, methanol).

Analysis for C26H31N305 • HCl • 0.7H20:

Calculated; C 60.68 H 6.41 N 8.16 Cl 6.88

Found: C 60.68 H 6.36 N7.95 Cl 6.48

Example 68:

(S) -l- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -4-phenylgly-cyl] -L-proline hydrochloride (20: 3)

a) 1- (bromoacetyl) -L-proline, 1,1-dimethylethyl ester

To a cooled (-10 ° C) solution of L-proline, 1,1-dimethylethyl ester (34.2 g, 0.2 mol) in methylene chloride (250 ml), diisopropylethylamine (38.3) is added. ml, 0.22 mol) and bromoacetyl chloride (16.5 ml, 0.2 mol), dropwise, over a period of 20 minutes, while maintaining the temperature between -10 and -5 ° C. The dark reaction mixture is stirred overnight (-10 ° C to room temperature) and concentrated under reduced pressure. The oily residue is redissolved in ethyl acetate, washed with saturated sodium bicarbonate (twice), 10% potassium bisulfate (twice) and with water (twice), dried (Na2SO4) and concentrated to a dark oily residue (28.0 g). By rapid chromatography (LPS-1 silica gel, 10% ethyl acetate / methylene chloride), 11.0 g of l- (bromoacetyl) -L-proline, 1,1-dimethylethyl ester are obtained under the form of a pale yellow oil.

b) 1- (N-phenylglycyl) -L-proline, 1,1-dimethylethyl ester

To a solution of 1- (bromoacetyl) -L-proline, 1,1-dimethylethyl ester (2.1 g, 7.5 mmol) in distilled tetrahydrofuran, aniline (1.5 g, 15 mmol) is added ) and the reaction mixture is stirred overnight under a nitrogen atmosphere. The reaction mixture is diluted with ethyl acetate (200 ml), washed with saturated sodium bicarbonate (2 x 50 ml) and with water (twice), dried (Na2SO4), and concentrated in vacuo to obtain a dark oily residue (4.0 g). By rapid chromatography (LPS-1 silica gel, 10% ethyl acetate / methylene chloride), 2.2 g of 1- (N-phenylglycyl) -L-proline, 1,1-dimethylethyl ester are obtained, in the form of a dark oil, which solidifies on drying under high vacuum.

c) (S) -1- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N-phenylgly-cyl] -L-proline, 1,1-dimethylethyl ester

Stirred at room temperature overnight under a nitrogen atmosphere, a reaction mixture containing 1- (N-phenyl-glycyl) -L-proline, 1,1-dimethylethyl ester (1.06 g, 3.5 mmol), (S) -N- [3-chloro-2-oxo-1- (phenylmethyl) propyl] benzamide (1.06 g, 3.5 mmol) from Example 63 (b), bicarbonate excess sodium, and sodium iodide (0.52 g, 3.5 mmol) in dimethylformamide (10 ml). The reaction mixture is poured into water (50 ml) and extracted with ethyl acetate (3 x 50 ml). The combined ethyl acetate extracts are washed with saturated sodium bicarbonate (twice) and water (three times), dried (Na2SO4) and concentrated under reduced pressure to give a dark oily residue (2.0 g ). By rapid chromatography (silica gel LPS-1, 5% ethyl acetate / methylene chloride to 15% ethyl acetate / methylene chloride), 0.3 g of (S) -1 is obtained. - [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N-phenylglycyl] -L-proline, 1,1-dimethylethyl ester, in the form of a pale yellow foam.

d) (S) -l- [N- [3- (benzoylamino) -2-oxo-4-phenylbutylJ-N-phenylgly-cyl] -L-proline, hydrochloride (20: 3)

The ester product from part (c) (0.28 g, 0.49 mmol) is treated with 2N hydrochloric acid / acetic acid. After stirring for 1 hour at room temperature, the reaction mixture is concentrated under reduced pressure, and the oily residue formed is triturated with ether (4 x) to give 0.14 g of (S) -l- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N-phenylglycyl] -L-proline, hydrochloride (20: 3), in the form of a whitish solid; m.p. 110-140 ° C; Rr 0.76 (low spot at 0.53) (silica gel, n-butanol / acetic acid / water, 3: 1: 1).

Analysis for C30H31N3O5 • 0.15HC1 • 0.5H20:

Calculated: C 68.22 H 6.13 N 7.95 Cl 1.00

Found: C 68.22 H 6.00 N 8.25 Cl 0.99

Examples 69 to 90:

By applying the method of Examples 63 to 68, but using the ketone indicated in column I, the acid chloride indicated in column II and the peptide ester indicated in column III, the ester product indicated in column 1 is obtained. column IV. By elimination of the Re ester group and other inhibiting groups, the corresponding final product is obtained in acid form.

Collar. I

H2N-CH-C-CH2C1

II

o

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659,475

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Collar. II

O

II

R2-C-Cl

Collar. III

r ri o

I I II

hn-ch-c-x (L)

Collar. IV

o r rî o

II I I II r3-ch-c-ch2-n-ch-c-x i (l)

NH

I

c = o I

R2

(Table on following pages)

The Rj inhibiting groups in Examples 74, 76 and 77, the R3 inhibiting groups in Examples 78 and 80, and the R5 inhibiting groups in Examples 81 and 83 to 85 are removed at the last stage of the synthesis. The R6 ester groups indicated in Examples 87 to 90 are not eliminated.

Example 91:

(+) -! - [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N-methyl-glycyl] -L-proline, monohydrochloride a) [3- (benzoylamino j -2-oxo- 4-phenylbutylJ methylcarbamic acid), phenylmethyl ester

N-methyl-N - [(phenylmethoxy) carbonyl] glycine (2.23 g, 10 mmol) is dissolved in 30 ml of tetrahydrofuran and cooled in an ice bath. Oxalyl chloride (1 ml, 11.5 mmol) is added, then 2 drops of dimethylformamide. After stirring for 30 minutes in an ice bath, the mixture is then stirred at room temperature for 1 hour. 0.25 ml of oxalyl chloride is added thereto. The mixture is then evaporated, redissolved in 15 ml of tetrahydrofuran and stirred in an ice bath. A solution of 2-phenyl-4- (phenylmethyl) -5- (4H) -oxazolone (3.1 g, 12.4 mmol) dissolved in 15 ml of tetrahydrofuran is added to the above-mentioned solution maintained under stirring in an ice bath . Triethylamine (1.4 ml, 10 mmol) is added and the solution is stirred at room temperature overnight. The precipitated triethylamine hydrochloride salt is filtered off. The tetrahydrofuran is separated from the residue then is redissolved in pyridine (5 ml) and p-dimethylaminopyridine (20 mg) is added. After stirring at room temperature for 3 hours, acetic acid (5 ml) is added and the reaction mixture is kept at 105 ° C for 30 minutes. The reaction mixture is then evaporated, the residue is dissolved in ethyl acetate and washed with aqueous sodium bicarbonate and with water. After trituration with ethyl acetate / hexane, 2.2 g of homogeneous acid [3- (benzoylamino) -2-oxo-4-phenylbutyl] methylcarbamic, phenylmethyl ester are obtained; m.p. 140-141 ° C.

b) (±) -N- [3- (methylamino) -2-oxo-l- (phenylmethyl) propyl] benza-mide, hydrochloride

Dissolve the acid [3- (benzoylamino) -2-oxo-4-phenylbutyl] methylcarbamic acid, phenyl methyl ester (0.5 g) in ethanol (50 ml) containing IN hydrochloric acid (2 ml ). A palladium on carbon catalyst (10%, 100 mg) is added and the hydrogenation is continued overnight. The reaction mixture is then filtered, evaporated, dissolved in water and lyophilized to 300 mg of (±) -N- [3- (methylamino) -2-oxo-1 - (phenylmethyl) propyl] benzamide, hydrochloride which occurs in the form of a homogeneous white powder.

c) (+) -l- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N-methylgly-cyl] -L-proline, 1,1-dimethylethyl ester

Stirred at room temperature, under a nitrogen atmosphere, overnight, a reaction mixture containing (±) -N- [3-îo (methylamino) -2-oxo-1 - (phenylmethyl) propyl] benzamide, hydrochloride ( 1.65 g, 5 mmol), 1- (bromoacetyl) -L-proline, 1,1-dimethylethyl ester (2.9 g, 10 mmol) from Example 68 (a) and di-isopropylethylamine (1.74 ml, 10 mmol) in dimethylformamide (20 ml). The reaction mixture is partitioned between water / ethyl acetate, and the aqueous layer is extracted again with ethyl acetate. The ethyl acetate extracts are dried (Na2SO4) and concentrated to a yellow oily residue (4.0 g). By instant chromatography (150 g of Merck silica 60 salt), 0.7 g of (±) -l- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N-methylglycyl] are obtained. -L-20 proline, 1,1-dimethylethyl ester, in the form of a yellow foam.

d) (+) -l- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N-methylgly-cyl) -L-proline, monohydrochloride

The ester product from part (c) (0.4 g, 0.79 mmol) is treated with 2N hydrochloric acid / acetic acid (5 ml). After stirring for 45 minutes at room temperature, the reaction mixture is concentrated under reduced pressure and the resulting oily residue is triturated with ether (3 x) to give 0.28 g of (+) - l- [N- [ 3- (benzoylamino) -2-oxo-4-phenylbutyl] -N-methylglycyl] -L-30 proline, monohydrochloride, in the form of a white solid; m.p. 125-130 ° C. Rf 0.73 (silica gel, n-butanol / acetic acid / water, 3: 1: 1).

Analysis for C2SH2pN3Os • HCl • 0.23H20:

35 Calculated: C 61.01 H 6.24 N 8.54

Found: C 61.01 H 6.10 N 8.36

Example 92:

Acid (S) -7 - [[[3- (benzoylamino) -2-oxo-4-phenylbutyl] methyl-40 aminoJacétyl] -l, 4-dithia-7-azaspiro [4.4] nonane-8-carboxylic, tri- fluoroacetate (1: 1)

a) (S) -7- (bromoacetyl) -1,4-dithia-7-azaspiro [4.4] nonane-8-carboxylic acid

45 (S) -1,4-dithia-7-azaspiro [4.4] nonane-8-carboxylic acid, hydrochloride (2.1 g, 8.7 mmol) is dissolved in IN sodium hydroxide (25 ml), cooled to 0 ° C. and bro-moacetyl bromide (2.1 g, 0.91 ml, 10.4 mmol) is added dropwise over a period of 10 minutes. The reaction mixture is stirred for 2 hours (from 0 ° C. to room temperature), washed with ethyl acetate (twice), and the aqueous layer is acidified to pH 2 and extracted with acetate. ethyl (three times). The assembled organic extracts are dried (Na2SO4) and concentrated to give 2.25 g of (S) -7- (bromoacetyl) -1,4-dithia-7-azaspiro [4.4] nonane-8-carboxyli-55 acid that .

b) (S) -7- (bromoacetyl) -1,4-dithia-7-azaspiro [4.4] nonane-8-carboxylic acid, diphenylmethyl ester

Dissolve in ethyl acetate (150 ml) (S) -7-60 (bromoacetyl) -1,4-dithia-7-azaspiro- [4.4] -nonane-8-carboxylic acid (2, 25 g, 6.9 mmol). Diphenyldiazomethane (1.3 g, 6.9 mmol) is added and the reaction mixture is stirred overnight at room temperature. The discolored reaction mixture is washed with saturated sodium bicarbonate (twice), 10% potassium bis-sulfate 65% (twice) and water, dried (Na2SO4), and concentrated to give 2.5 g (S) -7- (bromoacetyl) -1,4-dithia-7-azaspiro- [4.4] -nonane-8-carboxylic acid, diphenylmethyl ester, as a white solid residue.

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c) Acid (S) -7 - [[[3- (benzoylamino) -2-oxo-4-phenylbutyl] -methyl-aminoJacetyl] -1,4-dithia-7-azaspiro [4.4] nonane-8-carboxylic acid, di-phenylethyl ester

A mixture of the ester product from part (b) (2.4 g, 4.87 mmol), (±) -N- [3- (methylamino) -2-oxo-1 is stirred at room temperature overnight - (phenylmethyl) propyl] benzamide (0.81 g, 2.43 mmol) prepared as described in Example 91 (b) and diisopropylethylamine (0.85 ml, 4.87 mmol) in dimethylformamide (20 ml). The reaction mixture is poured into water (50 ml) and extracted with ethyl acetate (3 x 100 ml). The combined organic extracts are washed with saturated sodium bicarbonate, 10% potassium bisulfate and water, dried (Na2SO4) and concentrated to a dark oily residue (2.8 g). By simultaneous chromatography (200 g of Merck silica gel, 10% ethyl acetate / methylene chloride, 20% methanol / ethyl acetate), 1.1 g of (S) acid are obtained -

7 - [[[3- (benzoylamino) -2-oxo-4-phenylbutyl] methylamino] acetyl] -1,4-dithia-7-azaspiro [4.4] nonane-8-carboxylic, dimethylethyl ester in the form of a yellow oil.

d) Acid (S) -7- [[[3- (benzoylamino) -2-oxo-4-phenylbutyl] -methyl-amino] -acetylJ-1,4-dithia-7-azaspiro [4.4] nonane-8- carboxylic, trifluoroacetate salt (1: 1)

The ester product from part (c) (10.5 g, 0.72 mmol) is added to cooled trifluoroacetic acid (0 ° C) (2 ml) containing anisole (0.1 ml) . After stirring for 1 hour, the volatiles are removed in vacuo and the residue is treated with toluene (twice). The oily residue is triturated with ether (four times), which gives 0.36 g of (S) -7 - [[[3- (benzoylamino) -2-oxo-4-phenylbutyl] methylamino] acid] acetyl] -1,4-dithia-7-azaspiro [4.4] nonane-

8-carboxylic, trifluoroacetate salt (1: 1); m.p. 120-125 ° C. Rr 0.45 (spreading at fractionation) (silica gel, n-butanol / acetic acid / water, 4: 1: 1).

Analysis for C27H31N305S2 ■ C2HF302:

Calculated: C 53.11 H 4.92 N 6.40 S 9.78

Found: C 52.68 H 5.03 N 6.53 S 9.97

Example 93:

Acid (S) -2 - [[[3- (benzoylamino) -2-oxo-4-phenylbutyl] methyl-amino] acetyl] -1,2,3,4-tetrahydro-3-isoquinoline-carboxylic acid, monohydrochloride a) 2- (bromoacetyl) -1,2,3,4-tetrahydro-3-isoquinolinecarboxylic acid, 1,1-dimethylethyl ester

To a cooled solution of 1,2,3,4-tetrahydro-3-isoquinoli-necarboxylic acid, 1,1-dimethylethyl ester (7.7 g, 33 mmol) in methylene chloride (100 ml) is added diisopropylethylamine (6.23 ml, 36.3 mmol) and finally, over a period of 15 minutes, bromoacetyl bromide (6.6 g, 2.87 ml, 33 mmol) while maintaining the temperature at 5 ° C. The reaction mixture is stirred overnight (from -5 ° C. to room temperature), concentrated to approximately 33V3% of its volume, diluted with ethyl acetate (100 ml), washed with bicarbonate saturated sodium (twice), 10% potassium bisulfate (twice) and water (twice), dried (Na2SO4) and concentrated to a dark yellow semi-solid residue (11.0 g). By recrystallization from ethyl acetate / hexane, 4.2 g of 2- (bromoacetyl) -1,2,3,4-tetrahydro-3-isoquinolinecarboxyli-que, 1,1-dimethylethyl ester, are obtained. in the form of a solid cream-colored product; m.p. 92-95 ° C (85 ° C).

b) (S) -2 - [[[3- (Benzoylamino) -2-oxo-4-phenylbutyl] -methyl-amino] acetyl] -1,2,3,4-tetrahydro-3-isoquinolinecarboxylic acid, 1, 1-dimethylethyl ester

To a solution of 2- (bromoacetyl) -1,2,3,4-tetrahydro-3-isoquinolinecarboxylic acid, 1,1-dimethylethyl ester (2.12 g, 6 mmol) in dimethylformamide (20 ml), add (+) - N- [3- (me-

thylamino) -2-oxo-1- (phenylmethyl) propyl] benzamide (2.0 g, 6 mmol), prepared as described in Example 91 (b) and diisopropylethylamine (0.77 g, 1, 04 ml, 6 mmol). The reaction mixture is stirred overnight, poured into water (50 ml) and extracted with ethyl acetate (3 x 50 ml). The combined ethyl acetate extracts are washed with saturated sodium bicarbonate (twice) and water (twice), dried (Na2SO4) and concentrated to a yellow oily residue (2.9 g). By instant chromatography (200 g of Merck silica, 2% methanol / chloroform), 0.68 g of (S) -2 - [[[(3- (benzoylamino)) -2-oxo-4-phenylbutyl] acid is obtained. methylamino] acetyl] -1,2,3,4-tetrahydro-3-isoquinolinecarboxylic, 1,1-dimethylethyl ester, in the form of a desiccated foam of yellow color.

c) A cide (S) -2 - [[[3- (benzoylamino) -2-oxo-4-phenylbutylJmethyl-aminoJacetyl] -1,2,3,4-tetrahydro-3-isoquinolinecarboxylic acid, monohydrochloride

The ester product of part (b) (1.67 g, 1.17 mmol) is treated with 2N hydrochloric acid / acetic acid (5 ml). After stirring for 1 hour at room temperature, the reaction mixture is concentrated under reduced pressure and the oily residue formed is triturated with ether (four times) to give 0.55 g of acid (S) -2 - [[ [3- (benzoylamino) -2-oxo-4-phenyIbutyl] methylamino] acetyl] -1,2,3,4-tetrahydro-3-isoquinolinecarboxylic acid monohydrochloride in the form of a whitish solid; m.p. 123-126 C. Rf 0.47 (silica gel, n-butanol / acetic acid / water; 4: 1: 1).

Analysis for C30H31N3Os • HCl • 0.32H20:

Calculated: C 64.82 H 5.92 N 7.56 Cl 6.38

Found: C 64.82 H 6.22 N 7.55 Cl 6.13

Example 94:

[1 (+), 4S] -l- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N-methylglycyl] -4- (phenylthio) -L-proline, monohydrochloride a) (4S ) -l- (bromoacetyl) -4- (phenylthioj-L-proline

To a suspension of (4S) -4- (phenylthio) -L-proline (2.2 g, 10 mmol) in methylene chloride (50 ml, freshly distilled), bis (trimethylsilyl) acetamide (7, 35 ml, 30 mmol). The reaction mixture is stirred at room temperature for 2 hours until it becomes almost clear. The reaction mixture is then cooled to -53 ° C., and bromoacetyl chloride (1.9 g, 1.0 ml, 12 mmol) is added dropwise, maintaining the temperature at −5 ° C. After stirring overnight ( (-5C at room temperature), the reaction mixture is concentrated to approximately 50% of its volume, partitioned between saturated sodium bicarbonate / ethyl acetate, and the layers are separated. The organic layer is again extracted with saturated sodium bicarbonate. The combined aqueous layers are acidified to pH 2.0 with 10% potassium bisulfate and extracted with ethyl acetate (three times). The ethyl acetate fractions are combined, dried (Na2SO4) and concentrated to give 3.5 g of (4S) -1- (bromoacetyl) -4- (phenylthio) -L-proline, in the form of a viscous oil.

b) (4S) -1- (bromoacetyl) -4- (phenylthio J-L-proline, diphenylmethyl ester

A solution of diphenyldiazomethane (2.0 g, 10.2 mmol) in ethyl acetate (50 ml) is added dropwise to a solution of (4S) -l- (bromoacetate) -4- (phenylthio ) -L-proline (3.5 g, 10.2 mmol) in ethyl acetate (50 ml). The purple solution is stirred at room temperature overnight. The discolored reaction mixture is washed with saturated sodium carbonate (twice) and water (twice), dried (Na2SO4) and concentrated to give 4.78 g of (4S) -l- (bromoacetyl) -4 - (phenylthio) -L-proline, di-phenylmethyl ester, in the form of a yellow viscous oil.

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c) [1 (±), 4S] -l- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N-methylglycyl] -4- (phenylthio) -L-proline, diphenylmethyl ester

To a solution of (4S) -l- (bromoacetyl) -4- (phenylthio) -L-proline, diphenylmethyl ester (4.78 g, 9.4 mmol) in dimethylformamide (20 ml), (± ) -N- [3- (methylamino) -2-oxo-l- (phenylme-thyl) propyl] benzamide (2.42 g, 7.2 mmol), prepared as described in Example 91 (b), and diisopropylethylamine (0.93 g, 1.25 ml, 7.2 mmol). After stirring overnight at room temperature, the reaction mixture is poured into water (50 ml), and extracted with ethyl acetate (three times). The combined ethyl acetate extracts are washed with saturated sodium bicarbonate (twice) and water (twice), dried (Na2SO4), and concentrated to a yellow oil (6.2 g). By instant chromatography (Merck silica gel, 2% methanol / methylene chloride), 3.2 g of [1 (±), 4S] -1 - [N- [3- (benzoylamino) -2-oxo are obtained. -4-phenylbutyl] -N-methyl-glycyl] -4- (phenylthio) -L-proline, diphenylmethyl ester, in the form of a pale yellow foam.

d) [1 (±), 4S] -1- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N-methylglycyl] -4- (phenyltliio) -L-proline, monohydrochloride

The ester product of part (c) (1.6 g, 2.2 mmol) is treated with 2N hydrochloric acid / acetic acid (20 ml). After stirring for 2 hours at room temperature, the reaction mixture is concentrated under reduced pressure, and the oily residue is triturated with ether overnight, to give 1.2 g of [l (±), 4S] -l - [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N-methylglycyl] -4- (phenyl-thio) -L-proline, monohydrochloride, in the form of a whitish solid; m.p. 131-133 ° C; Rr0.38 (silica gel, n-butanol / acetic acid / water, 4: 1: 1).

Analysis for C31H33N305S • HCl • 0.9H20:

Calculated: C 60.82 H 5.90 N 6.87 S 5.24 Cl 5.79

Found: C 60.82 H 5.74 N6.94 S 5.25 Cl 5.75-

Example 95:

(4S) -l- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N-methyl-glycyl] -4- (4-fluorophenoxy) -L-proline, monohydrochloride a) (4S) -l- (bromoacetyl) -4- (fluorophenoxy) -L-proline, 1,1-dimethyl-ethyl ester

To a solution of (S) -4- (fluorophenoxy) -L-proline, 1,1-dimethyl ester (2.1 g, 7.5 mmol) in distilled tetrahydrofuran (50 ml), acid is added. bromoacetic (1.04 g, 7.5 mmol), hydroxybenzotriazole hydrate (1.14 g, 7.5 mmol) and dicyclohexylcarbo-diimide (1.54 g, 7.5 mmol). The reaction mixture is stirred overnight, the precipitated dicyclohexylurea is separated by filtration, and the filtrate is concentrated. The residue is dissolved in ethyl acetate (50 ml) and washed with saturated sodium bicarbonate (twice), 10% potassium bisulfate (twice) and water (twice), dried ( Na2SO4) and concentrated to give 3.1 g of (4S) -1- (bromoacetyl) -4- (fluorophenoxy) -L-proline, 1,1-dimethylethyl ester, as an oily residue.

b) (4S) -1- [N- [3- (benzoylamino) -2-oxo-4-phenylbutylJ-N-methyl-glycyl] -4- (fluorophenoxy j-L-proline, 1,1-dimethylethyl ester

To a solution of (4S) -1- (bromoacetyl) -4- (fluorophenoxy) -L-proline, 1,1-dimethylethyl ester (3.4 g, 8.5 mmol) in dimethylformamide (20 ml), add the (+) - N-3- (methylamino) -2-oxo-l- (phenylmethyl) -propylbenzamide (2.83 g, 8.5 mmol), prepared as described in Example 91 (b) and diisopropylethylamine (1.43 g, 1.92 ml, 11.0 mmol). After stirring overnight at room temperature, the reaction mixture is poured into water (100 ml) and extracted with ethyl acetate (three times). The combined ethyl acetate extracts are washed with saturated sodium bicarbonate (twice) with 10% potassium bisulfate (twice) and with water (twice), dried (Na2SO4) and concentrated to a residue dark (5.5 g). By simultaneous chromatography (silica gel LPS-1, 50% ethyl acetate / methylene chloride), 1.2 g of (4S) -1- [N- [3- (benzoylammo) -2-oxo are obtained -4-phenylbutyl] -N-methylglycyl] -4- (4-fluorophenoxy) -L-proline, 1,1-dimethylethyl ester, in the form of a pale yellow foam.

c) (4S) -1- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N-methylgly-cyl] -4- (4-fluorophenoxy) -L-proline, monohydrochloride

The ester product from part (b) (1.2 g, 1.95 mmol) is treated with 2N hydrochloric acid / acetic acid (20 ml). After stirring for 2 hours at room temperature, the reaction mixture is concentrated under reduced pressure, and the oily residue is triturated with ether overnight to give 0.92 g of (4S) -l- [N- [3 - (benzoylamino) -2-oxo-4-phenylbutyl] -N-methylglycyl] -4- (4-fluorophenoxy) -L-proline, monohydrochloride, in the form of a whitish solid; m.p. 131-140 ° C. Rf 0.54 (silica gel, n-butanol / acetic acid / water; 3: 1: 1).

Analysis for C31H32N3FOe • HCl • 0.55H20:

Calculated: C 61.24 H 5.65 N6.91 Cl 5.83

Found: C 61.24 H 5.66 N 7.09 Cl 5.70

Similarly, the procedure of Examples 91 to 95 can be used to prepare the compounds of Examples 1 to 90.

Example 96:

N- [N- [3- (benzoylamino) -2-oxo-4-phenylbutylJ-N-methylglycyl] -N-cyclohexylglycine, monohydrochloride a) (±) -N- [3- (benzoylamino) -2-oxo-4 -phenylbutyl] -N-methylgly-cine, 1,1-dimethylethyl ester

To a solution of (+) - N- [3- (methylamino) -2-oxo-l- (phenylme-thyl) propyl] benzamide (5.0 g, 15 mmol) prepared as described in Example 91 (b ) in dimethylformamide (20 ml), bromoacetic acid, 1,1-dimethylethyl ester (13.8 g, 3.15 ml, 19.5 mmol) and diisopropylethylamine (2.5 g, 3 are added) , 4 ml, 19.5 mmol). After stirring overnight at room temperature, the reaction mixture is poured into water (100 ml) and extracted with ethyl acetate (three times). The combined ethyl acetate extracts are washed with saturated sodium bicarbonate (twice), 10% potassium bisulfate (twice) and water (twice),

dried (Na2SO4) and concentrated in a yellow oil taking on the appearance of a desiccated foam after drying under high vacuum, to give 5.4 g of (+) - N- [3- (benzoylamino) - 2-oxo-4-phenylbutyl] -N-methylglycine, 1,1-dimethylethyl ester.

b) (±) -N- [3- (benzoylamino) -2-oxo-4-phenylbutylJ-N-methylgly-cine, hydrochloride

The ester product from part (a) (4.51 g, 11 mmol) is treated with 2N hydrochloric acid / acetic acid (20 ml). After stirring for 2.5 hours at room temperature, the reaction mixture is concentrated under reduced pressure and the oily residue is triturated with ether to give 3.3 g of (±) -N- [3- (benzoyl-amino ) -2-oxo-4-phenylbutyl] -N-methylglycyne, monohydrochloride in the form of a whitish solid.

c) N- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N-methylglycyl] -N-cyclohexylglycine, 1,1-dimethylethyl ester

To a solution of (±) -N- [3- (benzoylamino) -2-oxo-4-phenylbu-tyl] -N-methylglycine, hydrochloride (1.0 g, 2.6 mmol) in distilled tetrahydrofuran (50 ml), N-cyclohexylglycine, 1,1-dimethylethyl ester (0.55 g, 2.6 mmol), prepared as described in Example 65 (a), hydroxybenzotriazole hydrate (0, 39 g, 2.6 mmol) and dicyclohexylcarbodiimide (0.55 g, 2.6 mmol). The reaction mixture is stirred overnight, the precipitated dicyclohexylurea is separated by filtration, and the filtrate is concentrated. The residue is dissolved in ethyl acetate (50 ml) and washed with saturated sodium bicarbonate (twice), 10% potassium bisulfate (twice) and water (twice), dried ( Na2S04) and concentrated in one

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oily residue (1.5 g). By simultaneous chromatography (100 g, silica gel 60 Merck), 0.49 g of N- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N-methylglycyl] -N-cyclohexylglycine is obtained, 1,1-Dimethyl-ethyl ester in the form of foam.

d) N- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N-methylglycyl] -N-cyclohexylglycine, hydrochloride

The ester product from part (c) (0.48 g, 0.87 mmol) is treated with 2N hydrochloric acid / acetic acid (10 ml). After stirring for 2 hours at room temperature, the reaction mixture is concentrated under reduced pressure and the oily residue is triturated with ether overnight to give 0.32 g of N- [N- [3- (benzoylamino) - 2-oxo-4-phenylbutyl] -N-methylglycyl] -N-cyclo-hexylglycine, monohydrochloride, in the form of a whitish solid; m.p. 131-145 ° C. Rf 0.36 (silica gel, n-butanol / acetic acid / water, 4: 1: 1).

Analysis for C28H35N305 ■ HCl • 0.7H20:

Calculated: C 61.95 H 6.95 N 7.74 Cl 6.53

Found: C 61.95 H 6.74 N 7.71 Cl 6.23

Example 97:

Acid (S) -7 - ['[[(±) -3- (benzoylamino) -2-oxo-4-phenylbutyl] mé-thylamino Jacétyl] -l, 4-dithia-7-azaspiro [4.4Jnonane-8- carboxylic, methyl ester

To a solution of (+) - N- [3- (benzoylamino) -2-oxo-4-phenylbu-tyl] -N-methylglycine, monohydrochloride (1.0 g, 2.5 mmol) prepared as described in Example 96 (b), in distilled tetrahydrofuran (50 ml), (S) -1,4-dithia-7-azaspiro [4.4] nona-ne-8-carboxylic acid, methyl ester, monohydrochloride ( 0.66 g, 2.5 mmol), dicyclohexylcarbodiimide (0.54 g, 2.5 mmol), hy-droxybenzotriazole hydrate (0.39 g, 2.5 mmol) and diisopropylethylamine (0, 9 ml, 5 mmol). The reaction mixture is stirred overnight, the precipitated dicyclohexylurea is separated by filtration and the filtrate is concentrated. The residue is dissolved in ethyl acetate (100 ml) and washed with saturated sodium bicarbonate (twice) and with water (twice) dried (Na2SO4) and concentrated to a yellow oily residue (1.3 g). By simultaneous chromatography (Merck silica gel, 25% ethyl acetate / methylene chloride, 1% methanol / methylene chloride), 0.53 g of (S) -7 - acid is obtained - [[[( ±) -3- (benzoylamino) -2-oxo-4-phenylbutyl] methylamino] acetyl] -1,4-dithia-7-azaspiro [4.4] nonane-8-carboxylic, methyl ester, in the form of a foam white; m.p. 60-62 ° C. Rr 0.52 (silica gel, 5% methanol / methylene chloride).

Analysis for C28H33N30sS2 • 0.33H20:

Calculated: C 59.87 H 6.04 N7.48 S 11.42

Found: C 59.87 H 5.94 N7.56 S 11.36

Example 98:

Acid (S) -7 - [[[(±) -3- (benzoylamino) -2-oxo-4-phenylbutyl] mé-thy lamino] acétyl] -l, 4-dithia-7-azaspirof4.4] nonane- 8-carboxylic, methyl ester, monohydrochloride

The methyl ester product of Example 97 (0.26 g, 0.46 mmol) is treated with 2N hydrochloric acid / acetic acid until it becomes homogeneous (2 minutes), concentrated under reduced pressure , and the oily residue is triturated with ether (twice)

to give 0.26 g of acid (S) -7 - [[[(±) -3- (benzoylamino) -2-oxo-4-phenylbutyl] methylamino] acetyl] -1,4-dithia-7-azaspiro [4.4] nonane-8-carboxylic, methyl ester, monohydrochloride, in the form of a white solid; m.p. 79-85 ° C. Rf 0.53 (silica gel, 5% methanol / methylene chloride).

Analysis for C28H33N3OsS2 • HCl • 0.56H20:

Calculated: C 55.84 H 5.88 N6.98 S 10.64 Cl 5.88

Found: C 55.84 H 5.95 N6.78 S 10.43 Cl 5.66

Example 99:

(4S) -l- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N-methyl-glycyl] -4- (4-fluorophenoxy) -L-proline, methyl ester, monohydrochloride a) (4S) -4- (fluorophenoxy) -L-proline, methyl ester, monohydrochloride

To a suspension of (4S) -4- (phiorophenoxy) -L-proline (2.5 g. 11 mmol) in methanol at - 30 ° C, under an atmosphere of argon, thionyl chloride (8 , 09 ml, 11 mmol). The reaction mixture is stirred at -20 ° C for 2 hours, then at room temperature for 16 hours. The solvent is removed under reduced pressure and the residue is redissolved in methylene chloride (150 ml) and washed with IN sodium carbonate (twice) and with water (twice). After drying (MgSO 4), excess hydrochloric acid / methanol is added and the solvent is removed under reduced pressure. By addition of ether, a light brown solid (2.6 g) is obtained. By recrystallization from methanol / ether, 1.49 g of (4S) -4- (fluorophenoxy) -L-proline, methyl ester, monohydrochloride are obtained in the form of a light brown solid; m.p. 147-1481 C; [a] n = +6.96 (c = 1.55, methanol).

b) (4S) -1- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N-methyl-glycyl] -4- (4-fluorophenoxyJ-L-proline, methyl ester

To a solution of (+) -N- [3- (benzoylamino) -2-oxo-4-phenylbu-tyl] -N-methylglycine, monohydrochloride (1.17 g, 3 mmol) prepared as described in Example 96 (b), in distilled tetrahydrofuran (20 ml), is added (4S) -4- (fluorophenoxy) -L-proline, methyl ester, monohydrochloride (0.82 g, 3 mmol), hydroxybenzotriazole hydrate ( 0.46 g, 3 mmol) and dicyclohexylcarbodiimide (0.62 g, 3 mmol). The reaction mixture is stirred overnight, the precipitated dicyclohexylurea is separated by filtration and the filtrate is concentrated. The residue is dissolved in ethyl acetate (50 ml) and washed with saturated sodium bicarbonate (twice) and with water (twice), dried (Na2SO4) and concentrated to an oily residue (1.1 g). By instant chromatography (200 g, Merck silica gel 60; 3% methanol / chloroform), 0.15 g of (4S) -l- [N- [3- (benzoylamino) -2-oxo-4- is obtained. phenylbutyl] -N-methylglycyl] -4- (4- (fluorophenoxy) -L-proline, methyl ester, in foam form.

c) (4S) -1- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N-methylgly-cyl] -4- (fluorophenoxy) -2-proline, methyl ester, monohydrochloride

The methyl ester product of part (b) (0.15 g, 0.26 mmol) is treated with 2N hydrochloric acid / acetic acid until it becomes homogeneous (2 minutes), concentrated under pressure reduced, and the oily residue is triturated with ether (twice)

to give 0.14 g of (4S) -l- [N- [3- (benzoylamino) -2-oxo-4-phenyl-butyl [-N-methylglycyl] -4- (4-fluorophenoxy) -L-proline , methyl ester, monohydrochloride, in the form of a whitish solid; m.p. 105-125 ° C. Rr0.27 (silica gel, 5% methanol / chloroform).

Analysis for C32H34FO <; • HCl:

Calculated: C 62.79 H 5.76 N 6.86 F 3.10 Cl 5.79

Found: C 62.78 H 5.73 N 6.87 F 2.83 Cl 5.33

Example 100:

(4S) -l- [N- [(S) -3- (benzoylamino) -2-oxo-4-phenylbutyl] -L-alanyl] -4- (4-fluorophenoxy JL-proline, monohydrochloride a) (S) -N- [3- (benzoylamino) -2-oxo-4-phenylbutylJ-L-alanine, 1,1-dimethylethyl ester

To a stirred solution of (S) -N- [3-chloro-2-oxo-1- (phenylme-thyl) propylbenzamide (10.0 g, 33.1 mmol) in dimethylformamide (80 ml), L-alanine hydrochloride, 1,1-dimethylethyl ester (6.0 g, 33.1 mmol), sodium bicarbonate (6.1 g, 72 mmol) and sodium iodide (4.9 g, 33.1 mmol). The solution formed is stirred overnight at room temperature, poured into ether and washed with water (twice) and with bicarbonate.

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10% sodium. The ethereal solution is extracted with IN hydrochloric acid (three times), the combined extracts are made basic by the addition of solid sodium bicarbonate, then extracted with ethyl acetate (four times). The organic extracts are combined, dried (MgSO 4) and concentrated to give 8.9 g of a pale yellow solid. Part of this product is recrystallized from ethyl acetate to give (S) -N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -L-alanine, 1,1-dimethylethyl ester, as a white solid; m.p. 106.5-110 ° C.

b) (S) -N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -L-alanine, monohydrochloride

A solution of the ester product from part (a) (2.95 g, 5.4 mmol) in 1.4N hydrochloric acid in acetic acid (39 ml) is stirred at room temperature for 2 hours. The white precipitate formed is combined, rinsed with ether and dried to give 2.27 g of (S) -N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -L-alanine, monohydrochloride; m.p. 208-209 ° C (decomposition); [a] D = —71 ° (c = 0.38% in methanol). Rf (silica gel, chloroform / methanol / acetic acid, 4: 1: 1).

Analysis for C20H22N2O4 • HCl:

Calculated: C 61.64 H 5.93 N7.17 Cl 9.07

Found: C 61.33 H 5.97 N7.17 Cl 8.79

c) (S) -N- [N- (benzoylamino) -2-oxo-4-phenylbutyl] -N - [(phenylmé-thoxyjcarbonyl] -L-alanine

Triethylamine (2.1 ml, 15 mmol) is added to a mixture of (S) -N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -L-alanine, monohydrochloride (2.0 g , 5.1 mmol), benzyl chloroformate (730 ni, 5.1 mmol), water (7 ml) and dioxane (7 ml) at 25 ° C. The mixture formed is stirred at 25 ° C for 3 hours, then it is poured into an aqueous solution of sodium bicarbonate at 5% and washed with ether. The aqueous layer is acidified (HCl) and extracted into ethyl acetate (three times). The extract is dried (MgS04) and concentrated to give a colorless oil. By trituration with ether, a white granulated solid (150 mg) is obtained which is collected and set aside. The mother liquor is concentrated under vacuum to give 1.75 g of (S) -N- [N- (benzoylamino) -2-oxo-4-phenylbutyl] -N - [(phenylme-thoxy) carbonyl] -L-alanine , having the appearance of white glass.

d) (4S) -1- [N - [(S) -3- (benzoylamino) -2-oxo-4-phenylbutyl] -N- [(phenylmethoxy) carbonyl] -L-alanyl] -4- (4- fluorophenoxy) -L-proline, phenylmethyl ester

A mixture of (S) -N- [N- (benzoylamino) -2-oxo-4-phenylbutyl] -N - [(phenylmethoxy) carbonyl] -L-alanine (300 mg, 0.62 mmol), (4S) -4- (4-fluorophenoxy) -L-proline, phenylmethyl ester, p-toluenesulfonic acid salt (300 mg, 0.62 mmol), triethylamine (90 (il, 0.62 mmol), dicyclohexylcarbodiimide (130 mg , 0.62 mmol) and hydroxybenzotriazole hydrate (90 mg, 0.62 mmol) in tetrahydrofuran (7 ml) is stirred at 25 ° C for 20 hours, then the mixture is filtered and diluted with ethyl acetate The solution formed is washed successively with IN hydrochloric acid and a 10% aqueous solution of sodium bicarbonate, dried (MgSO 4), filtered and concentrated to give 500 mg of (4S) -1- [ N - [(S) -3- (benzoylamino) -2-oxo-4-phenylbutyl] -N - [(phenylmethoxy) car-bonyl] -L-alanyl] -4- (4-fluorophenoxy) -L-proline, phenylmethyl ester, in the form of a pale yellow oil.

e) (4S) -l- [N - [(S) -3- (benzoylamino) -2-oxo-4-phenylbutyl] -L-alanylJ-4- (4-fluorophenoxy) -L-proline, monohydrochloride

A mixture of the ester product from part (d) (500 mg, 0.6 mmol), a palladium on carbon catalyst (10%, 100 mg) of absolute ethanol (15 ml) and aqueous hydrochloric acid 1.0N (800 µl, 0.8 mmol) is hydrogenated under atmosphere and at 25 ° C for 17 hours, then is filtered and concentrated. The residue is chromatographed on HP-20, linear gradient from [9: 1, aqueous hydrochloric acid 0.0IN: methanol] to [1: 1, aqueous hydrochloric acid 0.01N: metha-

nol]. The fractions containing the desired product (TLC) are combined and concentrated. The residue is dissolved in a minimum amount of methanol. Ether is added, which forms a white precipitate, which is combined and dried under vacuum to give 200 mg of 5 (4S) -l- [N - [(S) -3- (benzoylamino) -2-oxo -4-phenylbutyl] -L-alanyl] -4- (4-fluorophenoxy) -L-proline, monohydrochloride; m.p. 152-153 ° C (decomposition); [a] "= —50 ° (c = 0.5, methanol). Rr 0.75 (silica gel, chloroform / methanol / acetic acid, 4: 1: 1).

Analysis for C31H32FN30 (s • HCl ■ 1.5H20:

Calculated: C 59.57 H 5.80 N 6.72 Cl 5.67

Found: C 59.68 H 5.56 N 6.67 Cl 5.99

Example 101:

15 [1 (S), 4R] -1- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -L-alanylJ-4-phenyl-L-proline, monohydrochloride a) (S) - N- [N- (benzoylamino) -2-oxo-4-phenylbutyl] -N - [(phenylmethoxy) carbonyl] -L-alanine, succinimido ester

A mixture of (S) -N- [N- (benzoylamino) -2-oxo-4-phenylbutylJ-N - [(phenylmethoxy) carbonyl] -L-alanine (800 mg, 1.6 mmol),

prepared as described in Example 100 (c), dicyclohexylcarbodiimide (340 mg, 1.6 mmol) and N-hydroxysuccinimide (190 mg, 1.6 mmol) in tetrahydrofuran (5 ml) is stirred at 25 ° C 25 for 18 hours. The product is then filtered and concentrated to give 950 mg of (S) -N- [N- (benzoylamino) -2-oxo-4-phenylbutyl] -N - [(phenylmethoxy) carbonyl] -L-alanine, succinimido ester.

b) [l (S), 4R] -1- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N-30 [(phenylmethoxy) carbonyl] -L-alanyl] -4-phenyl -L-proline

To a solution of (S) -N- [N- (benzoylamino) -2-oxo-4-phenylbu-tyl] -N - [(phenylmethoxy) carbonyl] -L-alanine, succinimido ester (950 mg, 1,6 mmol) in dimethylformamide (5 ml) is added (4R) -4-phenyl-L-proline, hydrochloride (375 mg, 1.7 mmol) and triethylamine (40 dj.1, 3.2 mmol ). The mixture formed is stirred at 25 ° C for 24 hours, then is poured into excess IN hydrochloric acid and is extracted with ethyl acetate (three times). The extracts are combined, dried (MgSO4), filtered and concentrated to give 1.1 g of [l (S), 4R] -l- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] - N-40 [(phenylmethoxy) carbonyl] -L-alanyl] -4-phenyl-L-proline.

c) [l (S), 4R] -1- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -L-alanyl] -4-phenyl-L-proline, hydrochloride

The product from part (b) (1.0 g, 1.5 mmol) and also contains ethanol (20 ml), water (5 ml), hydrochloric acid 1 0.01 (1.5 ml, 1.5 mmol) and a palladium on carbon catalyst (10%, 100 mg) is hydrogenated under the atmosphere and at 25 ° C for 18 hours, then is filtered and concentrated. The residue is chromatographed on HP-20 using a linear gradient of 0.01N aqueous chlorhy-50 dric acid: methanol, 40:60 to 10:90). The fractions containing the desired product (TLC) are combined and concentrated. The residue is dissolved in a minimum quantity of methanol. Ether is added and the white precipitate formed is combined and dried to give 300 mg of [1 (S), 4R] -1- [N- [3- (benzoylamino) -2-oxo-4-55 phenylbutyl] -L-alanyl] -4-phenyl-L-proline, monohydrochloride; m.p. 160-162 ° C (decomposition); [a] "= —57 ° (c = 1.5, methanol). Rf 0.8 (silica gel, chloroform / methanol / acetic acid, 4: 1: 1).

Analysis for C31H33N305 • HCl • 1.35H20:

60 Calculated: C 63.27 H 6.29 N7.14 Cl 6.02

Found: C 63.27 H 6.17 N 7.19 Cl 5.97

Example 102:

65 [1 (S), 4S] -1- [N- [3- (benzoylamino) -2-oxo-4-phenylbutylJ-L-alanyl] -4-phenyl-L-proline, hydrochloride

Based on the procedure of Example 101, but using the (4S) -4-phenyl-L-proline hydrochloride in the part

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(b), [1 (S), 4S] -l- [N- [3- (benzoylamino) -2-oxo-4-phenyl-butyl] -L-alanyl] -4-phenyl-L is obtained proline, monohydrochloride; m.p. 138-143 ° C;

[a] D = —61 ° (c = 0.3% in methanol). Rf 0.84 (silica gel, chloroform / methanol / acetic acid, 4: 1: 1).

Analysis for C31H33N305 • HCl • 2.13H20:

Calculated: C 61.80 H 6.35 N6.98 Cl 5.88 Found: C 61.80 H 6.06 N7.05 Cl 5.65

Example 103:

[1 (S), 4R] -! - [N- [3- (benzoylamino) -2 -oxo-4-phenylbutylJ-L-alanyl] -4-cyclohexyl-L-proline, hydrochloride

Based on the procedure of Example 101, but using the (4R) -4-cyclohexyl-L-proline hydrochloride in part (b), the [1 (S), 4R] -l is obtained - [N- [3- (benzoylamino-2-oxo-4-phenylbutyl] -L-alanyl] -4-cyclohexyl-L-proline, monohydrochloride; mp 140-154 ° C (decomposition);

[a] D = —83 ° (c = 0.36% in methanol). Rf 0.84 (silica gel, chloroform / methanol / acetic acid, 4: 1: 1).

Analysis for C31H39N305 • HCl ■ 0.79H20:

Calculated: C 63.71 H 7.17 N 7.19 Cl 6.06 Found: C 63.71 H 7.21 N 7.05 Cl 5.82

Example 104:

[1 (S), 4S] -l- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -L-alanyl] -4-cyclohexyl-L-proline, monohydrochloride

By following the procedure of Example 101, but using the (4S) -4-cyclohexyl-L-proline hydrochloride in part (b), we obtain [l (S), 4S] -l- [N-3- (benzoylamino) -2-oxo-4-phenylbutyl] -L-alanyl] -4-cyclohexyl-L-proline, hydrochloride; m.p. 139-141 ° C (decomposition);

[a] D = —80 ° (c = 0.2% in methanol). Rf 0.83 (silica gel, chloroform / methanol / acetic acid, 4: 1: 1).

Analysis for C3 jH ^ NjOs • HCl • 1.54H20:

Calculated: C 62.28 H 7.26 N7.03 Cl 5.93 Found: C 62.28 H 7.01 N7.02 Cl 6.16

Example 105:

Acid (S) -7 - [(S) -2 - [[3- (benzoylamino) -2-oxo-4-phenylbutyl] -amino] -l-oxopropylJ-1,4-dithia-7-azaspiro [4.4 Jnonane -8-carboxyli-que, monohydrochloride

By following the procedure of Example 101, but using the acid (S) -1,4-dithia-7-azaspiro [4.4] nonane-8-carboxyli-que, hydrochloride in part (b) as reactive product of L-proline, acid (S) -7 - [(S) -2 - [[3- (benzoylamino) -2-oxo-4-phenylbutyl] amino] -l-oxopropyl] -l is obtained , 4-dithia-7-azaspiro [4.4] nonane-8-carboxylic, monohydrochloride; m.p. 170-172 ° C; [a]! 5 = —26 ° (c = 1.4% in methanol). Rf 0.78 (silica gel, chloroform / methanol / acetic acid, 6: 1: 1).

Analysis for C27H31N30sS2 • HCl • 0.77H20:

Calculated: C 54.77 H 5.71 N7.10 S 10.83 Cl 5.99

Found: C 54.77 H 5.70 N 6.94 S 10.82 Cl 6.07

Example 106:

Acid [l (S), 5S] -1- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -L-alanyl] -4,5-dihydro-3-phenyl-1H-pyrazole- 5-carboxylic, monohydrochloride

By conforming to the operating mode of Example 101, but using the acid (S) -4,5-dihydro-3-phenyl-1 H-pyrazole-5-carboxy-

As the reacting product of L-proline, in part (b), the abovementioned compound is obtained.

Example 107:

(S) -2- [N - [(S) -3- (benzoylamino) -2-oxo-4-phenylbutyl] -L-alanylj-1,2,3,4-tetrahydro-3-isoquinolinecarboxylic acid monohydrochloride

By following the procedure of Example 101, but using the acid (S) -1,2,3,4-tetrahydro-3-isoquinoline-carboxyli-que hydrochloride, in part (b), to instead of the proline reagent, we obtain (S) -2- [N - [(S) -3- (benzoylamino) -2-oxo-4-phenylbutyl] -L-alanyl] -1,2,3 , 4-tetrahydro-3-isoquinolinecarboxylic, monohydrochloride.

Example 108:

Acid 1- [N - [(S) -3- (ben: oylamino) -2-oxo-4-phenylbutyl] -L-alanyl] -2- (2-hydroxyphenyl) -4 (Rj-thiazolidinecarboxylic acid, monohydrochloride

Following the procedure of Example 101, but using 2 - [(2-phenylmethoxy) phenyl] -4 (R) -thiazolidine-carboxylic acid, hydrochloride, in part (b), instead of the proline reagent, after elimination of the hydroxy inhibiting group, 1- [N - [(S) -3- (benzoylamino) -2-oxo-4-phenylbutyl] -L-alanyl] -2- ( 2-hydroxyphenyl) -4 (R) -thiazolidinecarboxyxlique, monohydrochloride.

Similarly, the methods of Examples 96, 97, 99 and 100 to 108 can be used to prepare the compounds of Examples 1 to 95.

Example 109:

(±) -l- [N- [3- (benzoylamino) -2-oxo-4-phenylbutylJ-L-alanylJ-L-proline, methyl ester, monohydrochloride a) l- [N- (2-ethoxy-2- oxoethyl) -N- [(phenylmethoxy) carbonylJ-L-alanyl] -L-proline, 1,1-dimethylethyl ester

The L-alanyl-L-proline, 1,1-dimethylethyl ester (22.44 g, 105 mol) is taken up in tetrahydrofuran (400 ml), while stirring. Bromoethyl acetate (11.64 ml, 105 mmol) and diisopropylethylamine (18.3 ml, 105 mmol) are added to this product. After 6 hours, the reaction mixture is cooled in an ice bath, and benzyl chloroformate (16.5 ml, 115.5 mmol) and diisopropylethylamine (20.1 ml, 115.5) are added. mmol). After 1 hour, the ice bath is removed, and the reaction mixture is kept overnight at room temperature. It is then concentrated to dryness, taken up in ethyl acetate, and washed to neutrality with 10% potassium bisulfate and saturated sodium bicarbonate. The crude product (52.3 g) is purified on a column of silica gel, eluting with ethyl acetate: hexane (1: 1) to give 42.4 g of 1- [N- (2- ethoxy-2-oxoethyl) -N - [(phenylmethoxy) carbonyl] -1-alanyl] -L-proline, 1,1-dimethylethyl ester.

b) l- [N- (carboxymethyl) -N- [(phenylmethoxy) carbonyl J-L-alanyl] -L-proline, 1,1-dimethylethyl ester

The ester product from part (b) (21 g, 45.4 mmol) is taken up in methanol (150 ml) and IN sodium hydroxide (50 ml, 50 mmol), while stirring at room temperature . After 5.5 hours, the methanol is removed in vacuo, and the aqueous portion is extracted with ethyl acetate. The aqueous portion is acidified with concentrated hydrochloric acid and extracted into ethyl acetate to give 16.95 g of l- [N- (carboxymethyl) -N - [(phenylmethoxy) -carbonyl] -L-alanyl ] -L-proline, 1,1-dimethylethyl ester.

c) (±) -l- [N- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N-

[(phenylmethoxy) carbonylJ-L-alanylJ-L-proline, 1,1-dimethylethyl ester

The ester product from part (b) (10.0 g, 23 mmol) is taken up in anhydrous tetrahydrofuran (75 ml) while stirring

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in an ice bath. To this product, oxalyl chloride (2.4 ml, 27.6 mmol) is added dropwise, followed by 15 drops of dimethylformamide. After 20 minutes, the ice bath is removed, and the reaction mixture is maintained at room temperature for 1 hour. The mixture is then concentrated to dryness under vacuum and taken up in tetrahydrofuran (40 ml), while stirring in an ice bath. To this product, 2-phenyl-4- (phenylmethyl) -5 (4H) -oxyzolone (6 g, 23.8 mmol) in tetrahydrofuran (35 ml) is added dropwise, followed by triethylamine ( 3.22 ml, 23 mmol). Additional triethylamine is added to maintain a basic atmosphere. After 20 minutes, the ice bath is removed and the reaction mixture is kept at room temperature overnight. The triethylamine hydrochloride is filtered off and the filtrate is concentrated to dryness under vacuum. The residue is taken up in pyridine (25 ml), 4-dimethylaminopyridine (75 mg) is added and the solution is stirred for 3 hours under an atmosphere of argon. Acetic acid (25 ml) is added and the reaction mixture is stirred for 45 minutes at 100 ° C under a positive stream of argon. The reaction mixture is concentrated to dryness, taken up in ethyl acetate and washed to neutrality with saturated sodium bicarbonate and dilute hydrochloric acid. The crude product (12.8 g) is chromatographed on silica gel, eluting with ethylerhexane acetate (1: 1) to give 9.05 g of (+) - l- [N- [N- [ 3- (benz-oylamino) -2-oxo-4-phenylbutyl] -N - [(phenylmethoxy) carbonyl] -L-alanyl] -L-proline, 1,1-dimethylethyl ester.

d) (±) -l- [N- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N- [(phenylmethoxy) carbonylJ-L-alanylJ-L-proline

The ester product from part (c) (11.4 g, 17.7 mmol) is taken up in trifluoroacetic acid (50 ml) and kept at room temperature for 45 minutes. The product is then concentrated to dryness and triturated with ether: hexane to give 10.3 g of crude product. The latter is purified on a column of silica gel in benzene / acetic acid (8: 2) to give 9.7 g of (+) - l- [N- [N- [3- (benzoylamino) -2-oxo -4-phenylbutyl] -N - [(phenylmethoxy) carbo-nyl] -L-alanyl] -L-proline; m.p. 70-91 ° C; [a] o = - 53.0 ° (c = 1.22, methanol). Rr0.41 (silica gel, benzene / acetic acid, 8: 2).

Analysis for C33H35N307:

Calculated: C 67.68 H 6.02 N7.17

Found: C 67.95 H 6.02 N6.82

e) (+) -! - [N- [N- [3- (benzoylamino) -2-oxo-4-phenylbutylJ-N - [(phenylmethoxy) carbonyl] -L-alanylJ-L-proline, methyl ester

The acid product of part (d) (3.5 g, 6 mmol) is taken up in dimethylformamide (12 ml) while stirring, at room temperature. Sodium bicarbonate (630 mg, 7.5 mmol) and methyl iodide (0.47 ml, 7.5 mmol) are added to this product. After 18 hours, additional methyl iodide (7.4 mmol) and sodium bicarbonate (7.4 mmol) are added. After stirring for an additional 4 hours, the reaction mixture is concentrated to dryness, under vacuum, taken up in ethyl acetate, and washed with saturated sodium bicarbonate. The crude product (3.7 g) is purified on a column of silica gel in ethyl acetate: hexane (2: 1) to give 3.5 g of (±) -l- [N- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N - [(phenylmethoxy) carbonyl] -L-alanyl] -L-proline, methyl ester.

f) (+) -! - [N- [3- (benzoylamino) -2-oxo-4-phenylbutylJ-L-alanylJ-L-proline, methyl ester, monohydrochloride

The ester product from (e) (900 mg, 1.5 mmol) is taken up in ethanol (95%, 100 ml) and IN hydrochloric acid (1.8 ml). A palladium on carbon catalyst (10%, 180 mg) is added and the reaction mixture is stirred under a hydrogen atmosphere, for 1 night. The reaction mixture is filtered to separate the catalyst, concentrated to dryness and lyophilized twice with water to give 700 mg of (±) -l- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -L-alanyl] -L-proline, methyl ester, monohydrochloride;

m.p. 110-128 ° C (90 ° C); [a] 2D3 = -69.9 ° (c = 1.18 methanol). Rr 0.63 (silica gel, chloroform / methanol / acetic acid, 9: 1: 1).

Analysis for C26H31N3Os • HCl • 0.92 H20:

Calculated: C 60.22 H 6.58 N8, ll Cl 6.84

Found: C 60.22 H 6.28 N8.10 Cl 7.06

Example 110:

(±) -! - [N-f 3- (benzoylamino) -2-oxo-4-phenylbutyl] -L-alanylJ-L-proline, methyl ester, monomethanesulfonate

(+) - 1 - [N- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N - [(phenylmethoxy) carbonyl] -L-alanyl] -L-proline, methyl ester ( 600 mg, 1 mmol), prepared as described in Example 109 (e), is taken up in methanol (60 ml) and methanesulfonic acid (0.066 ml, 1 mmol). A palladium on carbon catalyst (10%, 120 mg) is added, and the reaction mixture is stirred under a hydrogen atmosphere for 2.5 hours. The reaction mixture is filtered to separate the catalyst, then concentrated in vacuo. The crude product is triturated with ether and filtered. The precipitate is taken up in water and lyophilized to give 470 mg of (+) - l- [N- [3- (benzoyl-amino) -2-oxo-4-phenylbutyl] -L-alanyl] -L-proline , methyl ester, monomethanesulfonate; m.p. 80-140 ° C; [<x] d = —61.4 ° (c = 1.10, methanol). Rf 0.63 (silica gel, chloroform / methanol / acetic acid, 9: 1: 1).

Analysis for C26H31N305 • CH403S:

Calculated: C 56.98 H 6.34 N7.30 S 5.63

Found: C 56.98 H 6.32 N 7.47 S 5.63

Example 111:

(±) -l- [N- [3- (benzoylamino-2-oxo-4-phenylbutyl] -L-alanyl] -L-proline, methyl ester, hemisulfate

(+) - 1 - [N- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N - [(phenylmethoxy) carbonyl] -L-alanyl] -L-proline, methyl ester ( 600 mg, 1 mmol), prepared as described in Example 109 (e), is taken up in methanol (40 ml) with IN sulfuric acid (0.9 ml). A palladium on carbon catalyst (10%, 120 mg) is added and the reaction mixture is stirred under a hydrogen atmosphere for 2 hours. The reaction mixture is filtered to separate the catalyst, concentrated to dryness, triturated with ether and filtered. The crude product (446 mg) is taken up in water, filtered through a millipore filter, and lyophilized to give 400 mg of (±) -l- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -L-alanyl] -L-proline, methyl ester, hemisulfate; m.p. 98-112 ° C (85 ° C); [a] "= - 60.3 ° (c = 1.16, methanol). Rr 0.63 (during fractionation) (silica gel, chloroform / methanol / acetic acid, 9: 1: 1 ).

Analysis for C26H31N305 • 0.5H2S04 • 0.91H20:

Calculated: C 58.81 H 6.42 N7.92 S 3.01

Found: C 58.81 H 6.11 N7.91 S 3.09

Example 112:

(+) -! - [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -L-alanylJ-L-proline, propyl ester, monomethanesulfonate a) (±) -l- [N- [N - [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N - [(phenylmethoxy) carbonyl] -L-alanyl] -L-proline, propyl ester

La (±) -l- [N- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N - [(phenylmethoxy) carbonyl] -L-alanyl] -L-proline (1,17 g, 2 mmol), prepared as described in Example 109 (d), is taken up in tetrahydrofuran (2 ml) while stirring, in an ice bath. To this product, n-propanol (3.0 ml, 40 mmol) is added, then 4-dimethylaminopyridine (122 mg, 1 mmol) and dicyclohexylcarbodiimide (412 mg, 2 mmol). The reaction product is allowed to warm to room temperature, and the reaction is carried out for

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a night. The dicyclohexylurea is filtered off and the filtrate is concentrated to dryness under vacuum. The crude product (1.2 g) is chromatographed on a column of silica gel in benzene: acetic acid (9: 1) to give 1.0 g of (+) - l- [N- [N- [ 3- (benzoylamino) -2-oxo-4-phenylbutyl] -N - [(phenylmethoxy) carbonyl] -L-alanyl] -L-proline, propyl ester.

b) (±) -l- [N- [3- (benzoylamino) -2-oxo-phenylbutyl] -L-alanyl] -L-proline, propyl ester, monomethanesulfonate

The ester product from part (a) (500 mg, 0.8 mmol) is taken up in methanol (50 ml). Methanesulfonic acid (0.72 ml of an IN solution in methanol) and a palladium on carbon catalyst (10%, 100 mg) are added, and the reaction mixture is stirred under a hydrogen atmosphere for a night. The reaction mixture is filtered to separate the catalyst, concentrated to dryness, triturated with ether and filtered, to give 406 mg of crude product. This is taken up in water, filtered through a millipore filter and lyophilized to give 390 mg of (+) - l- [N- [3- (benzoy-lamino) -2-oxo-4-phenylbutyl] -L- alanyl] -L-proline, propyl ester, monomethanesulfonate; m.p. 82-94 ° C (69 ° C); [a] "= —60.5 ° (c = 0.95, methanol). Rf 0.46 (silica gel, chloroform / methanol / acetic acid, 90: 5: 5).

Analysis for C28H3SN305 • CH403S • 0.5H20:

Calculated: C 58.15 H 6.73 N7.01 S 5.34

Found: C 58.15 H 6.63 N7.05 S 5.34

Example 113:

(±) -l- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -L-alanylJ-L-proline, ethyl ester, monomethanesulfate a) (±) -l- [N- [N - [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N- [(phenylmethoxy) carbonyl] -L-alanyl] -L-proline, ethyl ester

La (±) -l- [N- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N - [(phenylmethoxy) carbonyl] -L-alanyl] -L-proline (1,17 g, 2 mmol), prepared as described in Example 109 (d), is taken up in tetrahydrofuran (2 ml), while stirring, in an ice bath. To the product are added absolute ethanol (2.3 ml, 40 mmol), then 4-dimethylaminopyridine (122 mg, 1 mmol) and dicyclohexylcarbodiimide (412 mg, 2 mmol). The reaction is allowed to take place overnight at room temperature. The dicyclohexylurea is separated by filtration, the filtrate is concentrated to dryness, the residue is taken up in ethyl acetate and washed to neutrality with 10% potassium bisulfate and saturated sodium carbonate. The crude product (1.2 g) is purified on a column of silica gel in benzene: acetic acid (8: 2) to give 1.0 g of (+) - l- [N- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N - [(phenylmethoxy) carbonyl] -L-alanyl] -L-proline, ethyl ester.

b) (±) -! - [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -L-alanylJ-L-proline, ethyl ester, monomethanesulfate

The ester product from part (a) (500 mg, 0.8147 mmol) is taken up in 50 ml of methanol-methanesulphonic acid methanolic (IN, 0.733 ml). A palladium on carbon catalyst (10%, 100 mg) is added, and the reaction mixture is stirred under positive hydrogen pressure for 3 hours. The reaction mixture is filtered to separate the catalyst, concentrated to dryness, triturated with ether and filtered. The precipitate (428 mg) is taken up in water,

filtered through a millipore filter, and lyophilized to give 360 mg of (±) -l- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -L-alanyl] -L-proline,

ethyl ester, monomethanesulfonate; m.p. 91-96 ° C (72 ° C); [a] o = —57.2 ° (c = 1.04, methanol). Rf 0.44 (silica gel, chloroform / methanol / acetic acid, 90: 5: 5).

Analysis for C27H33N3Os • CH4S03 • 0.6H20:

Calculated: C 57.33 H 6.56 N7.17 S 5.47

Found: C 57.33 H 6.43 N7.12 S 5.47

Example 114:

(+) -l- [N- [3- (benzoylamino) -2-oxo-4-phenylbutylJ-L-alanylJ-L-proline, 1-methylethyl ester, monomethanesulfonate a) (±) -l- [N- [ N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N-

[(phenylmethoxy) carbonylJ-L-alanylJ-L-proline, 1-methylethyl ester

La (±) -1 - [N- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N - [(phenylmethoxy) carbonyl] -L-alanyl] -L-proline (1,17 g, 2 mmol), prepared as described in Example 109 (d) and isopropanol (3.0 ml, 40 mmol) are taken up in tetrahydrofuran (2 ml), with stirring, in an ice bath . To this product, dicyclohexylcarbodiimide (412 mg, 2 mmol) is added. The reaction is allowed to take place overnight at room temperature. The product is then concentrated to dryness, taken up in ethyl acetate, and the dicyclohexylurea is separated by filtration. The filtrate is washed neutral with 10% potassium bisulfate and saturated sodium bicarbonate. The crude product (1.2 g) is purified on a column of silica gel with benzene: acetic acid (8: 2) to give 1.0 g of (±) -l- [N- [N-3- (benzoylamino ) -2-oxo-4-phenylbutyl] -N - [(phenylmethoxy) car-bonyl] -L-alanyl] -L-proline, 1-methylethyl ester.

b) (±) -! - [N- [3- (benzoylamino) -2-oxo-4-phenylbutylJ-L-alanylJ-L-proline, 1-methylethyl ester, monomethanesulfonate

The ester product from part (a) (815 mg, 1.3 mmol) is taken up in methanol (20 ml) and methanolic methanesulfonic acid (IN, 1.17 ml) and stirred under hydrogen in the presence of '' a palladium on carbon catalyst (10%, 160 mg) for 3 hours. The reaction mixture is filtered to separate the catalyst and concentrated to dryness under vacuum. The residue is triturated with ether, and the precipitate is filtered to give 704 mg of crude product. The latter is taken up in 35 ml of water (2% solution), filtered through a millipore filter and lyophilized to give 600 mg of (+) -l- [N- [3- (benzoylamino) -2-oxo-4- phenylbutyl] -L-alanyl] -L-proline, 1-methylethyl ester, monomethanesulfonate; m.p. 88-105 ° C; [a] o = - 55.2: (c = 1.05, methanol). Rr 0.33 (silica gel, chloroform / methanol / acetic acid, 9: 1: 1).

Analysis for C28H35N305 • CH403S • 0.66H20:

Calculated: C 57.89 H 6.75 N 6.99 S 5.33

Found: C 57.89 H 6.62 N 6.62 S 5.32

Examples 115-123:

By following the procedure of Examples 109, 112, 113 and 114, the (±) -l- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N - [(phenylmethoxy) carbonyl] -L-alanyl] -L-proline is treated with the reagent indicated below in column I. By eliminating the alanyl inhibiting group, the ester product indicated below in column II is obtained.

(Table on next page)

Example 124:

l- [N- [(S) -3- (benzoylamino) -2-oxo-4-phenylbutylJ-L-alanylJ-L-proline, sodium salt

1 - [N - [(S) -3- (benzoylamino) -2-oxo-4-phenylbutyl] -L-alanyl] -L-proline hydrochloride (424 mg, 1 mmol) is dissolved in water ( 50 ml). Aqueous sodium bicarbonate (0.1N, 20 ml) is added and the aqueous solution is lyophilized. It is then dissolved in water (10 ml) and the solution is applied to a column (5 cm x 60 cm) of Sephadex G-10 chromatographic gel and eluted with water. The fractions containing the desired product are combined and lyophilized, which makes it possible to obtain the sodium salt of l- [N - [(S) -3- (benzoylamino) -2-oxo-4-phenylbutyl] -L- alanyl] -L-proline.

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Collar. II

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1

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II

- CH2 - o — C — C (CH3) 3

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Br-CH2—0 — C — CH3

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-ch2-o-c-ch3

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1 II

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1 II

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1 day

1 li

-c — c-o-ch3

1 day

ch3

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-CH (CH2-OH) 2

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1 2 1 1 2

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-ch2-ch-ch2

| | S0 "®" a0 "®

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OH OH

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Preparation of pharmaceutical compositions Example 125:

1000 tablets each containing the following ingredients:

1 - [N - [(S) -3- (benzoylamino) -2-oxo-4- 5

phenylbutyl] -L-alanyl] -L-proline, sodium salt 100 mg wheat starch 50 mg gelatin 7.5 mg

Avicel (microcrystalline cellulose) 25 mg io magnesium stearate 2.5 mg are prepared from sufficiently large quantities by mixing the 1 - [N - [(S) -3- (benzoylamino) -2-oxo-4-phenylbutyl] -L-alanyl] -L-proline, sodium salt and wheat starch with an aqueous gelatin solution. The mixture is dried and ground into a fine i5 powder. Avicel, then magnesium stearate, are introduced and granulation is carried out. This mixture is then compressed in a tablet press to form 1000 tablets, each containing 100 mg of active ingredient.

Likewise, tablets containing 100 mg of the product of one of Examples 1 to 123 can be prepared.

An identical procedure can be used to form tablets containing 50 mg of active ingredient.

Example 126: 25

Two samples consisting of No. 1 gelatin capsules, each containing 50 mg of 1- [N - [(S) -3- (benzoylamino) -2-oxo-4-phenylbutyl] -L-alanyl] -L-proline , sodium salt, are filled with a mixture of the following ingredients:

1 - [N - [(S) -3- (benzoylamino) -2-oxo-4- 30

phenylbutyl] -L-alanyl] -L-proline,

sodium salt 50 mg magnesium stearate 7 mg lactose 193 mg

250 mg 35

Likewise, capsules containing 50 mg of the product of one of Examples 1 to 123 can be prepared.

Example 127:

A solution for injection is prepared as follows: 40

l- [N - [(S) -3- (benzoylamino) -2-oxo-4-phenylbutyl] -L-alanyl] -L-proline,

sodium salt 500 g methyl paraben 5 g propyl paraben 1 g sodium chloride 25 g water, for injection 5 1

The active substance, stabilizers and sodium chloride are dissolved in 3 liters of water for injection, then the volume is brought to 5 liters. The solution is filtered through a sterile filter in order to asep-tically fill the pre-sterilized vials which are then closed with pre-sterilized rubber stoppers. Each vial contains 5 ml of solution at a concentration of 100 mg of active ingredient per ml of solution for injection.

In the same way, a solution for injection containing 100 mg of active ingredient per ml of solution can be prepared for the product obtained by one of examples 1 to 123.

Example 128:

1000 tablets each containing the following ingredients: l- [N - [(S) -3- (benzoylamino) -2-oxo-4-phenylbutyl] -L-alanyl] -L-proline,

sodium salt

100 mg

Avicel

100 mg hydrochlorothiazide

12.5 mg lactose

113 mg wheat starch

17.5 mg stearic acid

7 mg

370 mg are prepared from sufficiently large quantities, by making lozenges containing l- [N - [(S) -3- (benzoylamino) -2-oxo-4-phenylbutyl] -L-alanyl] -L-proline, sodium salt, Avicel and part of stearic acid. The pellets are ground and passed through a No. 2 sieve, then mixed with hydrochlorothiazide, lactose, wheat starch and the rest of the stearic acid. The mixture is compressed into tablets in the form of 350 mg capsules in a tablet press. These tablets are notched to allow them to be divided into halves.

Similarly, tablets containing 100 mg of the product of one of Examples 1 to 123 can be prepared.

R

Claims (2)

659,475 CLAIMS 1. Compound having the formula: - (CH2) r — OH, - (CH,) r — NH2, - (CH2) r — SH, - (CH2) rS-lower alkyl, (L) ORR, O II II R3-CH-C-CH2-N — CH-CX I NH IC = 0 I r2 y1 HO - (CH2) r — NH-C, or - (CH2) r — C — NH2, NH2 “provided that R1 is hydrogen only when R is other than hydrogen; or a pharmaceutically acceptable salt of this compound, formula in which: X is 15 R'esi (R14> p '• Wi' .- O. CH- y ^ H2? I 8 r CH-9 r »ST CH_ H, C CH 2i I -N C-COOR, '-N C-COOR, t -N - C-COOR, / (L) H R10 \ / Rio | (L) H' X (L) Rllv _ S. / R12 25 H0C CH. 21 i 2 n -2T7. R3 is hydrogen, lower alkyl,, R14> p '11' '12 -N C- COOR, -N C-COOR, / -N C-COOR, / cl) (l) | 6 a halo-substituted lower alkyl, - (CH2) m-cycloalkyl, O 30 "(CH2) OH, - (CH2) - OH C® '-N n fr - (CH2) r — OH, - C-COOR, / -N — C-COOR, / -N C-COOR, t (L) 24 (L) (L) (CH2} ririj '40 N -N - C-COOR,, -N C-COOR, / -N 1 | It C-COOR- '-N— C-COOR- / -N C-COOR- / -CH-0-C-R18, -C — C-0-R23s -CH- (CH2-OH) 2, I (L) 6 | (L) 6 | (L) 6 He H H H Rl7 ^ -22 ~ CH2 — CH — ÇH2, - (CH2) 2 - N (CH3) 2, or -CHj {o); 10 fo y le; òh òh n Ri 7 is hydrogen, lower alkyl, cycloalkyl or phen- c-COOR / -N) or - N - CH - COOR6; | (L) 6 ^ C-COORg 'I I Rj8 is hydrogen, lower alkyl, lower alkoxy, 15 d I (L) R * Rs _ _. h phenyl, or R17 and R18 combined are - (CH2) 2 -, - (CH2) 3 -, —CH = CH, or w R7 is hydrogen, lower alkyl, halogen, keto, hydroxy, alkyl O R24 is hydrogen, lower alkyl, * "\ 0 (R14} p ' —NH — C - lower, azido, amino, - € ?. -He. - -tf). R21 and R22 are independently selected from the group consisting of hydrogen and lower alkyl, and R23 is lower alkyl. 1 (L) I (L) 6 C-COOR, H H I (L) -N — CH — COORe; NOT I H - (CH2) r-NH2, - (CH,) r — SH, - (CH2) r-S-loweralkyl, -JH O R is hydrogen, lower alkyl, cycloalkyl, - <OUU <0> - (CH2) 2-NH2, - (CH2) 3-NH2, - (CH2) 4-NH2, - (CH2) 2 — OH, - (CH2) 3 — OH, - (CH2) 4-OH, - (CH2) 2-SH, - (CH2) 3 —SH, or - (CH2) 4-SH; R! is hydrogen, lower alkyl, halo-substituted lower alkyl, - (CH2) r - <^ Q - (CH2) r — Q ^> - OH, - (CH2) r-NH-C ^ or - (CH2) r — C — NH2; \ h2 R4 is hydrogen, lower alkyl, ~ (CH2) iT {5) '- (CH2) m-cycloalkyl, "(CH2) dd-fTj), Rs is hydrogen, lower alkyl, - (cH2) F1j- |, H OH •OH - (CH_) 2 r NOT OH I H H 3 659,475 - (ch2} - T N Here > KT NOT, •NOT I H - (CH2) r — OH, - (CH2) r — NH2, - (CH2) r — SH, - (CH2) r-S-lower alkyl, Rs is a keto, a halogen. O RI5 ene, - O —C —NT Ru lower alkyl-O, (R13'p NH O - (CH2), - NH-, or - (CH2) r — C — NH2; NH2 R is an integer from 1 to 4; R7 is hydrogen, lower alkyl, halogen, keto, hydroxy, alkyl or a 1- or 2-naphthyloxy of the formula: -O- (CH_) -2 nor lower S-alkyl Of (R14) p. O - NH - C— lower, azido, amino, N, / \ , R19 (r R, -nh-c- (ch2) 20 or a 1- or 2-naphthylthio of formula: -s- (CH_) -2 m (E14> p ' 13 p ' (R14> p < -ICHj) (R13'p ' = o- R0 is a keto group or - (CH ^) - (R13, p ' 30 - (ch ~) a 1- or 2-naphthyl of formula: not - (CH2) m-cycloalkyl, Rj 0 is halogen or - Y - R16; Ru, R'n, R12 and R ', 2 are independently selected from hydrogen and lower alkyl or R' ,,, R12 and R'12 are hydrogen, and RM is (R14) p5 <R14'p ' o R1S -O — C — NT, lower alkyl-O, Ris -o- (ch2) (R13} p ' a 1- or 2-naphthyloxy of formula: 2 m lower S-alkyl, 40 R13 is hydrogen, lower alkyl of 1 to 4 carbon atoms, lower alkoxy of 1 to 4 carbon atoms, lower alkyl thio of 1 to 4 carbon atoms, chloro, bromo, fluoro, trifluoromethyl, hydroxy, phenyl, phenoxy, phenylthio or phenyl-methyl; 45 R14 is hydrogen, lower alkyl of 1 to 4 carbon atoms, lower alkoxy of 1 to 4 carbon atoms, lower alkyl thio of 1 to 4 carbon atoms, chloro, bromo, fluoro, trifluoromethyl or hydroxy; m is zero, one, two, three or four; so p is one, two or three, provided that p is greater than one, only if R13, or Rj4, is hydrogen, methyl, methoxy, chloro or fluoro; Rls is hydrogen or lower alkyl of 1 to 4 carbon atoms; 55 Y is oxygen or sulfur; R16 is lower alkyl of 1 to 4 carbon atoms. -s- (ch2) (R13'p or a 1- or 2-naphthylthio of formula: -s- (ch2) or the R16 groups joined to complete an unsubstituted 5 or 6 membered nucleus, or said nucleus in which one or more of the 65 carbon atoms have a lower alkyl of 1 to 4 carbon atoms or a substituent di (lower alkyl of 1 to 4 carbon atoms); R19 is lower alkyl, benzyl or phenethyl; 659,475 R20 is hydrogen, lower alkyl, benzyl or phenethyl; R6 is hydrogen, lower alkyl, benzyl, benzhydryl, O not 0 R21 O 5 -N J 11 11 2. Pharmaceutical compositions containing at least one compound of formula (I) according to claim 1 as a pharmacologically active ingredient. a 1- or 2-naphthyl of formula: