BE897327A - SUBSTITUTED PEPTIDE COMPOUNDS AND THEIR INTERMEDIATES, THEIR PREPARATION AND THEIR USE - Google Patents

Abstract

Composed of substituted peptides of formula (I), in which the different symbols have different meanings, these compounds being found to be of interest as hypotensive agents and, depending on the definition of X, as analgesics, and process for the preparation of these compounds.

Description

       

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   DESCRIPTIVE MEMORY filed in support of a request for
PATENT OF INVENTION formed by
E. R. SQUIBB & SONS, INC. for: "Substituted peptide compounds and their intermediates, their preparation and their use".



  Priority of a patent application in the United States of America filed on July 19, 1982, under No. 399,650 in the name of Sesha I. Natarajan and Eric M. Gordon.
 EMI1.1
 



  ----------------------------------------------------------

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 "Substituted peptide compounds and their intermediates, their preparation and their use".



   The present invention relates to substituted peptide compounds of formula I, and to the salts of these compounds:
 EMI2.1
 formula in which X represents an amino or imino acid of the formula:
 EMI2.2
 

  <Desc / Clms Page number 3>

 
 EMI3.1
 
 EMI3.2
 represents hydrogen, a lower alkyl radical, a halogen, a keto, hydroxy radical,
 EMI3.3
 o il - lower, azido, amino, -N, -

  <Desc / Clms Page number 4>

 
 EMI4.1
 0 Il -NH-C- 13 2-9 (R) -2-C il 14 p (R 13) r. 4 - m -, - (CH,), ---.-, 0 - (CH 2) m j, - (CH2) mf:

   NOT A WORD' - <CH2) m &commat;, o s
 EMI4.2
 a 1-or 2-naphthyl group of the formula
 EMI4.3
 - (CH) -, -) -cycloalkyle, m 0 is / - \ -) - \ '15
 EMI4.4
 a group 1-or the formula
 EMI4.5
 -), -S-'lower alkyl, <'p -) - or a group l-or 2-naphthyl-' - 'p'
 EMI4.6
 of the formula:
 EMI4.7
 - 2 MJ) 0

  <Desc / Clms Page number 5>

 
 EMI5.1
 R8 represents a keto radical, a halogen, o -O-C-N R15 - a lower radical, (R13) l3'p
 EMI5.2
 a 1-or 2-naphthyloxy group of the formula
 EMI5.3
 2m ... ze (R14) (R). C) 14 p - (R13)
 EMI5.4
 or a 1-or 2-naphthylthio group of the formula
 EMI5.5
 - 2m t R9 represents a keto radical or.



  M (R13)
 EMI5.6
 represents a halogen or-Y-R-,.



  - LO-Lb R-, R ', RetR're independently represent a lower alkyl hydrogen atom or R', R and R'-. hydrogen and Rll represents
 EMI5.7
 horn - <4'p

  <Desc / Clms Page number 6>

 
R7 R13 represents hydrogen, a lower alkyl radical of 1 to 4 carbon atoms, a lower alkoxy radical of 1 to 4 carbon atoms, a lower alkylthio radical of 1 to 4 carbon atoms, a chloro, bromo radical, fluoro, trifluoromethyl, hydroxy, phenyl, phenoxy, phenylthio or phenylmethyl.



  R14 represents hydrogen, a lower alkyl radical of 1 to 4 carbon atoms, a lower alkoxy radical of 1 to 4 carbon atoms, a lower alkylthio radical of 1 to 4 carbon atoms or a chloro, bromo, fluoro radical , trifluoromethyl or hydroxy. m is zero, one, two, three or four. p is equal to one, two or three on the condition that p is greater than one only in the case where R13 or R14 represents hydrogen or a methyl, methoxy, chloro or fluoro radical.



   R15 represents hydrogen or a lower alkyl radical of 1 to 4 carbon atoms.
 EMI6.1
 



  Y represents oxygen or sulfur.
 EMI6.2
 



  R16 represents a lower alkyl radical of 1 to 4 carbon atoms or- (R13) (R 13) p
 EMI6.3
 or alternatively the R groups join to form an unsubstituted pentagonal or hexagonal ring, this cycle possibly comprising a lower alkyl substituent of 1 to 4 carbon atoms or a di substituent (lower alkyl of 1 to 4 carbon atoms) on one or more of the its carbon atoms.

  <Desc / Clms Page number 7>

 
 EMI7.1
 R4 represents hydrogen, an alkyl radical
 EMI7.2
 lower, - 0, - (CH) m- - f, -, z '"S 0
 EMI7.3
 
 EMI7.4
 R5 represents hydrogen, lower alkyl, - \ r
 EMI7.5
 
 EMI7.6
 - (CH -) - SH, - (CH) -S-lower alkyl,
 EMI7.7
 o o - (CH-NH-C) -C-NH NH
 EMI7.8
 r is an integer from 1 to 4.



      (CH2) - R19 represents a lower alkyl, benzyl or phenethyl radical, R20 represents hydrogen or a radical

  <Desc / Clms Page number 8>

 
 EMI8.1
 lower alkyl, benzyl or phenethyl.



  R represents hydrogen or a lower alkyl radical, '- (CH2), -,' 24'2 '- (CH) -OH, - (CH) -OH, -) -OH, -) -SH, - ) -SH) -SH.



  Ru of hydrogen, a lower alkyl radical, a lower alkyl radical substituted by halo or a group
 EMI8.2
 
 EMI8.3
 - (CH-OH, -) -S-- lower, 0.



  - (CH -) - NH-C -, NH
 EMI8.4
 under the condition ceRt represents hydrogen only in the case where R is different from hydrogen.

  <Desc / Clms Page number 9>

 
 EMI9.1
 R2 represents- {CH2}; &commat; , - 'C'm- c - CCH2) fj or. m 0
 EMI9.2
 represents hydrogen, a lower alkyl radical,
 EMI9.3
 - '.



  (R14) p s - (CH -) - t) - Oj, a lower alkyl radical 2.



  -) -cycloalkyle, -) Y-OH, r OH - (CH2) '-' 2 H 1.



  H - (CH) -NH, - (2r '' '2r' '' lower alkyl -) -NH-C 2 (2) NH2

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 cycloalkyl, - (CH2) where m, R-, P and r are as defined above.



   R6 represents hydrogen, an alkyl radical
 EMI10.1
 lower, benzyl unradical, benzhydryl radical
 EMI10.2
 o 0 or a group il 1 il - C-O-R 1 R17 R22 - 2 2 1 2 OH OH
 EMI10.3
 hydrogen or a lower alkyl, cycloalkyl or phenyl radical. represents hydrogen or a J-O lower alkyl, lower alkoxy or phenyl radical, or else R17 and represent
 EMI10.4
 
 EMI10.5
 R- and hydrogen or a lower alkyl radical.



   R21 R23 represents a lower alkyl radical.



   The present invention, in its broadest aspect, relates to the substituted peptide compounds of formula I above, to the compositions containing these compounds and to the method of using these compounds as pharmaceutical agents, as well as to the intermediates used in the preparation of these compounds.



    The expression "lower alkyl" used in the definition of the various symbols refers to straight or branched chain radicals containing up to 7 carbon atoms. Preferred lower alkyl groups have up to 4 carbon atoms, with methyl and ethyl being the most

  <Desc / Clms Page number 11>

 favorite. Similarly, the terms "lower alkoxy" and "lower alkylthio" refer to lower alkyl groups of this type attached to an oxygen or sulfur atom.



   The term "cycloalkyl" refers to saturated rings having from 3 to 7 carbon atoms, the cyclopentyl and cyclohexyl groups being the most preferred.



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



     The term "halo substituted lower alkyl" refers to the aforementioned lower alkyl groups in which one or more hydrogens have been replaced by chloro, bromo or fluoro groups, such as trifluoromethyl, which is preferred, pentafluoroethyl, 2, 2, 2-trichloro-
 EMI11.1
 ethyl, chloromethyl, bromoethyl, etc.
 EMI11.2
 



  The symbols - l - - 2m- - 0 N "'0 2 m, 0 J show that the alkylene bridge is attached to an available carbon atom.



   The compounds of formula I can be prepared, in which R represents hydrogen,
 EMI11.3
 by converting a carboxymethyl peptide ester
 EMI11.4
 of the formula: \ (II) HO-C-CH-N-CH-C-X Il 2 (L) 0 R40

  <Desc / Clms Page number 12>

 in its acid chloride and then by reaction with an oxazolone of the formula:
 EMI12.1
 
 EMI12.2
 formulas in which] and R6 (in the definition of X) represent protecting groups such as, for example, a benzyloxycarbonyl group for
R40 and a benzyl group for R6. Separation of the protective groups R40 and R, for example by hydrogenation, gives the products of formula I, in which R6 represents hydrogen.



   The carboxymethyl peptide ester of formula II is prepared by reacting the ester of
 EMI12.3
 peptide of the formula: R Il (IV) H-N-CH-C-X 2
 EMI12.4
 with tert-butyl bromoacetate and then by introducing the protecting group, for example by treatment with benzyl chloroformate.



   The compounds of formula I can also be prepared by reacting a ketone of the
 EMI12.5
 formula: 0 0 Il (V) R-C-NH-CH-C-CH - halo 212 R 3 3 in which halo represents Cl or Br with the peptide ester of the formula:

  <Desc / Clms Page number 13>

 
 EMI13.1
 in the presence of a base, such as sodium bicarbonate, this reaction being followed by separation
 EMI13.2
 from the ester group 6
A6. We can prepare the intermediate acetone of formula V by treating a ketone of the formula:
 EMI13.3
 in which R40 represents a protecting group, such as benzyloxycarbonyl, with bromide. hydrogen- and acetic acid, this reaction being followed by a reaction with the acid halide of the formula:
 EMI13.4
 in the presence of a base, such as sodium bicarbonate.



   The compounds can also be prepared
 EMI13.5
 of formula I by reacting an aminoketone of
 EMI13.6
 the formula: Il (IX) R-C-NH-CH-C-CH-NH 2 1 2 R R in particular the hydrochloride thereof, with the ester
 EMI13.7
 of aminoacetyl amino or imino acid of the formula:
 EMI13.8
 

  <Desc / Clms Page number 14>

 in which R6 in the definition of X represents an easily separable ester protective gauze and halo represents Cl or Br.



   The compounds of formula l can also be prepared by coupling an aminoketone carboxylic acid of the formula:
 EMI14.1
 with the amino or imino acid ester of the formula: (XII) HX in the presence of a coupling agent, such as dicyclo-
 EMI14.2
 hexylcarbodiimide, in the definition of X representing an easily separable protection group.



  The separation of the protection group R6 from the o products of formula I, in which R6 represents hydrogen.



   We can prepare aminoketone from
 EMI14.3
 formula IX by converting carboxyalylamine from
 EMI14.4
 the formula: 1 (XIII) HO-C-CH-N-R, Ii 2 40 o 0
 EMI14.5
 in which represents a protective group, such as a benzyloxycarbonyl group, in its acid chloride and then by reaction with an oxazolone of formula III to give a compound of the formula:
 EMI14.6
 

  <Desc / Clms Page number 15>

 Separation of the protection group R40 ′, for example by hydrogenation, gives the reagent of formula IX.



   The aminoketone of formula IX, in which R is different from hydrogen, can also be prepared by reacting the ketone of formula V with a substituted amine of the formula: (XV) R-NH2
The aminoketone carboxylic acid of formula XI can be prepared by reacting the aminoketone of formula IX with a haloacetic acid ester of the formula:
 EMI15.1
 
 EMI15.2
 in which Prot represents an easily separable ester protection group, such as the t-butyl group,
 EMI15.3
 to give the ester of formula II (XVII) R-C-NH-CH-C-CH-NH-CH-C-O-Prot 2 (L) R3
 EMI15.4
 Separation of the ester protecting group gives the reagent of formula XI.



   In the above reactions, if any or all of R, R., and R5 'represent
 EMI15.5
 

  <Desc / Clms Page number 16>

 
 EMI16.1
 then the hydroxyl, amino, imidazolyl, mercaptan or guanidinyl function must be protected during the reaction. Interesting protecting groups are benzyloxycarbonyl, t-butoxycarbonyl, benzyl, benzhydryl, trityl, etc., and nitro in the case of the guanidinyl group. The protecting group is separated by hydrogenation, treatment with an acid, or by any other known method after the completion of the reaction.



   We can chemically treat the
 EMI16.2
 ester lines of formula I, in which 6 represents a lower alkyl, benzyl or benzhydryl group, with sodium hydroxide in aqueous dioxane or with trimethylsilyl bromide so as to obtain the products of formula I, in which R6 represents hydrogen.



  Benzyl and benzhydryl esters can also be hydrogenated, for example, by treatment with hydrogen in the presence of a palladium on carbon catalyst.



   The products formed of esters of formula l in which R6 represents:

  <Desc / Clms Page number 17>

 
 EMI17.1
 can be obtained using the peptide of formula IV or VI or the haloacetyl amino or imino acid ester of formula X in the above-mentioned reactions with this ester group already in place. These reagents formed from esters can be prepared by treating the peptide of formula IV or VI or the haloacetylamino-ouimino acid ester of formula X, in the-
 EMI17.2
 which R represents hydrogen, with a chloride
 EMI17.3
 acid such as: il o ou (H
 EMI17.4
 so as to protect the atom of N.

   The protected compound is then reacted in the presence of a base with
 EMI17.5
 a compound of the formula 0 II (XVIII) L-CH-0-C-R-- <-L R17
 EMI17.6
 wherein L is a leaving group such as chlorine, bromine, tolylsulfonyl, etc., this reaction being followed by the separation of the protecting group from N, such as by treatment with an acid or by hydrogenation.



   It is also possible to obtain the ester products of formula I, in which R represents
 EMI17.7
 

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 by treatment of the product of formula I, in which R6 represents hydrogen, with a molar excess of the compound of formula XVIII.



   The ester products of formula I can be prepared, in which R represents
 EMI18.1
 
 EMI18.2
 ctj-L-. LU-muj -'- j.
Lct. Lucmc. m- a j-'j. uuuj-L. uc j. ct R6 represents hydrogen, with a molar excess of the compound of the formula:
 EMI18.3
 We can prepare ester products
 EMI18.4
 of formula I, in which R b
 EMI18.5
 
 EMI18.6
 by coupling the product of formula I, in which R6 of hydrogen, with a molar excess b of the compound of formula:
 EMI18.7
 or the formula:
 EMI18.8
 in the presence of a coupling agent, such as dicyclohexylcarbodiimide, this reaction being followed by the separation of the hydroxyl protecting groups.

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 EMI19.1
 



  Esters of formula I in which a lower alkyl group can be obtained from the carboxylic acid compounds, i.e. the compounds in which hydrogen represents 6, by esterification processes streams, for example by treatment with an alkyl halide of the formula or an alcohol of the b formula R-OH.



  6
R6 We can obtain the peptide esters of formulas IV and VI by coupling the hydrochloride salt of the amino ester or imino acid with the formula
 EMI19.2
 XII, in which, for example, a benzyl group, with the amino acid N protected from the
 EMI19.3
 
 EMI19.4
 in which prot represents a protection group,
 EMI19.5
 such that 0 il - reaction is carried out
 EMI19.6
 preferably in the presence of a coupling agent, such as dicyclohexylcarbodiimide. Separation of the protecting group from N, for example by treatment with trifluoroacetic acid, gives the peptide esters of formulas IV and VI.



   The aminoacid or imino acid haloacetyl ester of formula X can be prepared by reacting the aminoacid imino acid ester of formula XII with a haloacetyl halide of formula:
 EMI19.7
 

  <Desc / Clms Page number 20>

 
The products of formula I in which R7 represents an amino group can be obtained by reducing the corresponding products of formula I in which R7 represents an azido group.



   The preferred compounds of this
 EMI20.1
 invention with regard to the peptide part of the structure of formula l are those in which:
R represents hydrogen, a lower straight or branched chain alkyl group of 1 to 4 carbon atoms or a phenyl group.



     R-represents hydrogen, a lower straight or branched chain alkyl group of 1 to 4 atoms'
 EMI20.2
 carbon, -CF, - (CH -) - NH r represents an integer from 1 to 4,
 EMI20.3
 - O) -CHO-OH, -CH - OH, 2 2 OH -CT'TO''T "" NHHH NH - (CH -) - S-CH., - 2 2 J 2 J 2 NH 0 0 It - the condition
 EMI20.4
 that R only represents hydrogen in the case where R is different from hydrogen.



   R4 represents hydrogen or a cyclohexyl or phenyl group.

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   R5 represents hydrogen or a lower alkyl group with a straight or branched chain of 1 to 4 carbon atoms, -CH OH,
 EMI21.1
 
R6 represents hydrogen, a lower straight or branched chain alkyl group from 1 to 4
 EMI21.2
 carbon atoms, an alkali metal salt,
 EMI21.3
 ö Il - CH-O-C-R -, - CH-C-OR.



  1 R17.



  R17 - (CH-OH) or -CH-CH --- CH 2 1 1 OH OH
0 R23 represents a lower alkyl group with a straight or branched chain of 1 to 4 carbon atoms, in particular the group -C (CH3) 3.



     R17 represents hydrogen, an alkyl group

  <Desc / Clms Page number 22>

 lower than a straight or branched chain of 1 to 4 carbon atoms or a cyclohexyl group.
 EMI22.1
 R18 an alkyl group lower than j-0 represents a straight or branched chain of 1 to 4 carbon atoms or a phenyl group.



   R represents a lower alkyl group with a straight or branched chain of 1 to 4 carbon atoms, in particular the group C (CH).



   R7 represents hydrogen.



   R7 represents hydroxy.



   R7 represents a lower straight or branched chain alkyl group of 1 to 4 carbon atoms or a cyclohexyl group.



   R7 represents an amino group.



   R7 represents a group -0- lower alkyl in which the lower alkyl is a straight chain or
 EMI22.2
 branched with 1 to 4 carbon atoms.
 EMI22.3
 



  R 7 3 M 13
 EMI22.4
 where m is zero, one or two and R13 is hydrogen or methyl, methoxy, methylthio, chloro, bromo, fluoro or hydroxy.
 EMI22.5
 



  R represents - -m-- (13
 EMI22.6
 an l-naphthyloxy or 2-naphthyloxy group in which m is equal to zero, one or two and R13 represents hydrogen or a methyl, methoxy, methylthio, chloro, bromo, fluoro or hydroxy group.



     R7 represents a group-S-lower alkyl

  <Desc / Clms Page number 23>

 in which the lower alkyl is a straight or branched chain containing from 1 to 4 carbon atoms.



   R represents
 EMI23.1
 or a l-naphthylthio or 2-naphthylthio group, in which m is zero, one or two and R13 represents hydrogen or a methyl, methoxy, methylthio, chloro, bromo, fluoro or hydroxy group.
 EMI23.2
 



  R8 is a lower group o in which the lower alkyl is a straight or branched chain containing from 1 to 4 carbon atoms.



   RQ represents
 EMI23.3
 in which m is zero, one or two and R13 represents hydrogen or a methyl, methoxy, methylthio, chloro, bromo, fluoro or hydroxy group.
 EMI23.4
 represents a group-S-lower alkyl 0 in which the lower alkyl is a straight or branched chain containing from 1 to 4 carbon atoms.
 EMI23.5
 



  R8 represents - \ 2m R13
 EMI23.6
 in which m is zero, one or two and represents hydrogen or a methyl, methoxy, methylthio, chloro, bromo, fluoro or hydroxy group.



     R represents a phenyl, 2-hydroxyphenyl or 4-hydroxyphenyl group.
 EMI23.7
 



  LesR a fluoro or chloro group. The R-repésQ-itent a group-Y-R-, wherein 10 lu Y represents 0 or S, and an alkyl

  <Desc / Clms Page number 24>

 
 EMI24.1
 less than a straight or branched chain of 1 to 4 carbon atoms or the R16 groups are linked to form an unsubstituted pentagonal or hexagonal ring, this cycle possibly comprising a methyl or dimethyl substituent on one or more of the available carbons. and all hydrogen or Ril a phenyl, 2-hydroxyphenyl or 4-hydroxyphenyl group and R '-, R and hydrogen.



  The most interesting compounds of the present invention with regard to the peptide part of the structure of the formula are those in which:
 EMI24.2
 X represents-N-CH - COOR-, 1 R4 / Ry R7 s s 2 H '-N - C-COOR, -N - C-COOR H2C t (L) 6' (L) 6 H H t
 EMI24.3
 

  <Desc / Clms Page number 25>

 
 EMI25.1
 R represents hydrogen or methyl. R-represents hydrogen, methyl or - (CH) NH, in particular methyl, on the condition that only represents hydrogen if R is different from hydrogen.



   R6 represents hydrogen, a straight or branched chain lower alkyl of 1 to 4 carbon atoms or an alkali metal salt.



   R4 represents a cyclohexyl or phenyl group.



   R7 represents hydrogen, a cyclohexyl group, a lower alkoxy group from 1 to 4
 EMI25.2
 carbon atoms, - (CH.) - / \ 2 R13 - - - R13
 EMI25.3
 where m is zero, one or two, and represents hydrogen or a methyl, methoxy, methylthio, Cl, Br, F or hydroxy group, the compounds of the invention being particularly advantageous when R7 represents hydrogen . t is equal to two or three, in particular two.



   Compounds of interest of the present invention with respect to the keto portion of the
 EMI25.4
 structure of formula I, are those in which
 EMI25.5
 R2 represents - 2'm-O R R14

  <Desc / Clms Page number 26>

 where m is zero, one or two and R14 represents hydrogen or a methyl, methoxy, methylthio, Cl, Br, F or hydroxy group, in particular a phenyl group.
 EMI26.1
 



  R3 represents a lower alkyl group
 EMI26.2
 with straight or branched chain from 1 to 4 carbon atoms, - (CH,), - NH-m --- mR14 ---. 1--, s 1-R14 ----, - -N
 EMI26.3
 L'-t., I /) -c - Lc-se-ijLuc. c hydrogen or a methyl, methoxy, methylthio, Cl, Br, F or hydroxy group, and r is an integer from L to 4, in particular a benzyl group.



   The compounds of formula I, in which
 EMI26.4
 R6 of hydrogen, form salts with b a series of inorganic or organic bases. Pharmaceutically acceptable, non-toxic salts are preferred, although other salts are also useful for isolating or purifying the product. These pharmaceutically acceptable salts are metal salts, such as sodium, potassium or lithium salts, alkaline earth metal salts, such as calcium or magnesium salts, or salts derived from amino acids, such as arginine, lysine, etc. The salts are obtained by reaction of the acid form of the compound with an equivalent of the base bringing the desired ion in a medium in which the salt precipitates or in an aqueous medium and then by lyophilization.



   Similarly, the compounds of formula I, in particular in which R6 represents an ester group, form salts with a series

  <Desc / Clms Page number 27>

 inorganic and organic acids. Again, non-toxic pharmaceutically acceptable salts are preferred, although other salts are also useful for isolating or purifying the product. These pharmaceutically acceptable salts are those obtained with hydrochloric acid,. methanesulfonic acid, sulfuric acid, maleic acid, etc. The salts are obtained by reacting the product with an equivalent amount of acid in a medium in which the salt precipitates.



   As noted above, the peptide portion
 EMI27.1
 the molecule of the products of the formula l R Ri 0 represented by: 1 1 Il - (L) is in the configuration L. An asymmetric center is also present in the keto portion of the molecule when R3 is different from the hydrogen.



  Thus, the compounds of formula I can exist in the form of diastereoisomers or mixtures of diastereoisomers. Racemates, enantiomers or diastereoisomers can be used in the methods described above as starting materials. When preparing diastereoisomeric products, they can be separated by ordinary chromatographic or fractional crystallization methods.



   The products of formula I, in which the imino acid cycle is monosubstituted, gives rise to a cis-trans isomerism. The configuration of the final product
 EMI27.2
 will depend on the configuration of the substituent R, and 7 o

  <Desc / Clms Page number 28>

    R redans the starting material of formula XII.



   The compounds of formula I, and their pharmaceutically acceptable salts, are hypotensive agents. They inhibit the conversion of the angiotensin I decapeptide to angiotensin II and, therefore, they are useful for reducing or eliminating hypertension linked to angiotensin. The action of the enzyme renin on angiotensinogen, a pseudoglobulin in the blood, produces angiotensin 1.



  Angiotensin I is converted by the angiotensin converting enzyme (ACE) to angiotensin II. The latter is an active hypertensive agent which has been considered to be the causative agent in several forms of hypertension in various species of mammals, for example in humans. The compounds of the present invention are involved in the angiotensinogen-Y (resin) -anciotensin I-; - angiotensin II sequence by inhibiting the angiotensin converting enzyme and by reducing or eliminating the formation of the hypertensive agent angiotensin II. Thus, by administering a composition containing one (or a combination) of the compounds of the present invention, angiotensin-dependent hypertension is eliminated in a mammal (for example in a human being) suffering from this type hypertension.



   A single dose, or preferably two to four divided daily doses, given on the basis of about 0.1 to 100 mg, preferably about 1 to 50 mg, per kg of body weight per day, are found to be suitable for reducing blood pressure.

  <Desc / Clms Page number 29>

 



  The substance is preferably administered orally, but parenteral routes can also be used, such as the subcutaneous, intramuscular, intravenous or intraperitoneal routes.



   The compounds of the present invention can also be formulated in combination with a diuretic for the treatment of hypertension.



  A combination product comprising a compound of the present invention and a diuretic can be administered in an effective amount which comprises a total daily dosage of about 30 to 600 mg, preferably about 30 to 330 mg of a compound of the present invention, and from about 15 to 300 mg, of about 15 to 200 mg of the diuretic, in a mammal that requires it.

   Examples of diuretics which can be used in conjunction with a compound of the present invention are thiazide-type diuretics, for example chlorothiazide, hydrochlorothiazide, flumethiazide, hydroflumethiazide, bendroflumethiazide, methyclothiazide, trichloromethiazide, polythiazide , or benzthiazide as well as ethacrynic acid, ticrynafene, chlorthalidone, furosemide, musolimine, bumetanide, triamterene, amiloride and spironolactone, as well as the salts of these compounds.



   The compounds of formula I may be formulated to reduce blood pressure in the form of compositions, such as tablets, capsules or elixirs for oral administration, or in the form of sterile solutions or suspensions

  <Desc / Clms Page number 30>

 for parenteral administration. Approximately 10 to 500 mg of a compound of formula I is combined with a physiologically acceptable vehicle, carrier, excipient, binder, preservative, stabilizer, flavoring, etc. in a unit dosage form that meets product standards pharmaceutical. The amount of active substance in these compositions or preparations is such that an appropriate dosage is obtained within the range indicated.
 EMI30.1
 



  The compounds of formula I, in which X
 EMI30.2
 represents-NH-CH-COOR-, also have a 1 5
 EMI30.3
 enkephalinase inhibiting activity and are useful as analgesic agents. Thus, by administering a composition containing a compound of formula I or a combination of compounds of formula I or a pharmaceutically acceptable salt of these compounds, pain can be eliminated in a mammal. A single dose, or preferably two to four divided daily doses, given on the basis of about 0.1 to about 100 mg per kg of body weight per day, preferably from about 1 to about 50 mg per kg per day, produce the desired analgesic activity.

   The composition is preferably administered by the oral route, but parenteral routes can also be used, such as the subcutaneous route.



   The following examples illustrate the invention.



  Temperatures are given in degrees Celcius. LH-20 is a Sephadex chromatography gel commercially available from Pharmacia Fine Chemicals.

  <Desc / Clms Page number 31>

 



   Example 1 1- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -L-alanyl] -L-proline hydrochloride. a) Salt of p-toluenesulfonic acid of phenyl-methyl ester of L-alanyl-L-proline.



   N- [(1,1-dimethylethoxy) carbonyl] -L- alanine (310.7 g), L-proline phenylmethyl ester hydrochloride (396.2 g), dicyclohexylcarbodiimide (338.6 g) , hydroxybenzotriazole hydrate (251.5 g), diisopropylethylamine (285.7 ml) and tetrahydrofuran (5 liters) are combined with OOC (dicyclohexylcarbodiimide is added last) and stirred overnight at Room temperature. The reaction mixture is filtered and concentrated. The residue is dissolved in 4 liters of ethyl acetate and washed with 5% sodium bicarbonate (2 x 2 liters), 5% potassium bisulfate (2 x 2 liters) and water . The layer is dried with ethyl acetate (MgSO 4) and concentrated.



  The residue is dissolved in 3 liters of diethyl ether and left overnight at OOC. The mixture is filtered and the filtrate is concentrated, giving 620 g of crude N- [(1,1-dimethylethoxy) '- carbonyl] -L-alanyl-L-proline phenylmethyl ester.



   N- [(1,1-dimethylethoxy) carbonyl] -L-alanyl-L-proline phenylmethyl ester (275 g) is cooled in an ice bath and treated with 500 ml of cold trifluoroacetic acid ( freshly distilled) under argon and stirred at room temperature for one hour. The mixture is concentrated under vacuum and brought twice to an azeotrope state.

  <Desc / Clms Page number 32>

 with toluene. The crude trifluoroacetic acid salt, which looks like an oil, is dissolved in 500 ml of ether and treated slowly with stirring with a solution of p-toluenesulfonic acid hydrate (1 equivalent) dissolved in 6 liters of ether. The resulting precipitate is collected by filtration. The solid is dissolved in 1 liter of methanol and treated with 4 liters of ether and cooled.

   The resulting precipitate is collected, dried and 300 g of p-toluenesulfonic acid salt of L-alanyl-L-proline phenyl ester are obtained; melting point of 1560-1580C. b) 1- [N- [2 (1,1-dimethyl-ethoxv) -2-oxoethyl] -N- [(phenylmethoxy) carbonyl] -L-alanyl] -L-proline phenylmethyl ester.



   A mixture of p-toluenesulfonic acid salt of phenylmethyl ester of L-alanyl-L-proline (32.16 g; 72 mmol), tert-butyl bromoacetate (14, 14) is stirred at ambient temperature for 72 hours. 2 g; 72 mmol), triethylamine (20.1 ml, 144 mmol) and tetrahydrofuran (290 ml). 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 this water and ethyl acetate. the materials 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 (0.062 mm - 0.037 mm) using the solvent system formed

  <Desc / Clms Page number 33>

 
 EMI33.1
 ethyl acetate and hexane (1/1) (pressure of 0, MPa), which allows to obtain 19.5 g of phenylmethyl ester of 1- [N- [2- 2-oxoethyl] -N- proline. c) 1- [N- (N-carboxymethyl) -N- [(phenylmethoxy) carbonyl] -L-alanyl-L-'proline phenylmethyl ester.



   Phenylmethyl ester of
 EMI33.2
 1-methoxy) carbonyl-L-alanyl] -L-proline (18 g; 34.3 mmol) in trifluoroacetic acid (50 ml) and left to stand at room temperature for 1.5 hours.



  The mixture is evaporated and filtered through a small column of silica gel (200 gr) using the chloroform / methanol / acetic acid solvent mixture (9.6 / 0.2 / 0.2) to give 11.4 gr of 1- [N- (N-carboxymethyl) -N- [(phenylmethoxy) -carbonyl] - L-alanyl-L-proline phenylmethyl ester. d) 1- [N- [3- (benzovlamino) -2-oxo-4-phenylbutyl] -N- [(phenylmethoxy) carbonyl] -L-alanyl] L-proline phenylmethyl ester.



   1- [N- (carboxymethyl) -N - [(phenylmethoxy) carbonyl] -L-alanyll-L-proline phenylmethyl ester (7.0 g; 15.0 mmloes) is dissolved in dry tetrahydrofuran (50 ml), it is cooled in an ice bath, and oxalyl chloride (1.57 ml; 18 mmol) is added followed by 4 drops of dimethylformamide.



  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

  <Desc / Clms Page number 34>

 in an ice bath. The solution is added dropwise over 5 minutes to a solution of 2-phenyl-4- (phenylmethyD-5 (4H) -oxazolone (3.96 g; 15.75 mmol) in tetrahydrofuran (24 ml) cooled in ice 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 evaporated. the filtered tetrahydrofuran solution and redissolved in pyridine (16 ml).



  4-Dimethylaminopyridine (50 mg) is added and the solution is stirred at room temperature for 3 hours. Glacial 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 under vacuum, dissolved in ethyl acetate and extracted with aqueous sodium bicarbonate and dilute hydrochloric acid.

   The ethyl acetate extract after evaporation is chromatographed on silica gel (0.062 mm-0.037 mm) using the ethyl acetate / benzene solvent system (4/6) so as to obtain 4.9 gr of phenylmethyl ester
 EMI34.1
 than 1- [N- [3- [Analysis: calculated for C40H41N307. 0.42 H20:
VS   ; 70, 31; N: 6.15; H: 6.17 Found C: 70.31; N: 6.13; H: 6.08 e) 1- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -L-alanyl] -L-proline hydrochloride.



   Phenylmethyl ester of

  <Desc / Clms Page number 35>

   1- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N- [(phenylmethoxy) carbonyl] -L-alanyl] -L-proline (1.0 g) in absolute ethanol (30 ml) and in IN hydrochloric acid (2.25 ml). Palladium on carbon catalyst (10%; 200 mg) is added and the solution is stirred under a hydrogen atmosphere overnight. The catalyst is filtered off and the solution is evaporated. The crude product (700 mg) is combined with 300 mg crude product from a previous small-scale reaction and passed through an LH-20 column (2.54 cm x 38.1 cm) in methanol . The fractions containing the desired product are combined, evaporated, dissolved in water and filtered.

   The clear aqueous solution is lyophilized and 800 mg of 1- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -L-alanyl] -L- monohydrochloride are obtained.
 EMI35.1
 Proline; [at] ; 6 (c = 1.4, methanol); melting point of 98-130 C (decomposition).



  Thin layer chromatography (silica gel; n-butanol / acetic acid / water; 4/1/1) Rf = 0.44.



  Analysis: calculated for C25H30N305Cl. 0.75 H2O:
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 1- [N- [7-amino-3- (benzoylamino) -2-oxoheptvl] -L-alanvl] -L-proline dihydrochloride. a) N2-benzoyl-N6 - [(phenylmethoxy) carbonyl] -L-lysine.



   Benzoyl chloride (5 ml; 43.2 mmol) and aqueous sodium hydroxide (4N; 10.8 ml) are added simultaneously in 5 over a period of 30 minutes to a solution of N6¯ [phenyl-

  <Desc / Clms Page number 36>

 methoxy) carbonyl] L-lysine (10.09 gr; 36mmol) in de. aqueous sodium hydroxide (1N; 26 ml) cooled in ice. The ice bath is removed and stirring is continued for an additional 1.5 hours at room temperature. The reaction mixture is then extracted with ethyl acetate (which is discarded), the aqueous mother liquor is acidified with dilute hydrochloric acid and extracted with ethyl acetate .



  The extract is concentrated with ethyl acetate and the residue is crystallized from a mixture of ethyl acetate and hexane so as to obtain 12.9 g of N-benzoyl-
 EMI36.1
 vs ; melting point of 110 -112 C (1090C). b) 2-Phenyl-4- [4- [[(phenylmethoxy) carbonyl] amino] - butyl] -5 (4H) -oxazolone.



   N2-benzoyl-N6- [(phenylmethoxy) carbonyl] -L-lysine (11.53 g; 30 mmol) is dissolved in tetrahydrofuran (55 ml) and stirred in an ice bath. To this reaction mixture is added dropwise over a period of 15 minutes a solution of dicyclohexylcarbodiimide (6.8 g; 33 mmol) in tetrahydrofuran. The ice bath is removed after 1 hour and stirring is continued at room temperature for an additional 18 hours.



  The dicyclohexylurea is filtered off and the tetrahydrofuran is concentrated in vacuo. the residue is crystallized from a mixture of ethyl acetate and
 EMI36.2
 of hexane and 9.4 g of 2-phenyl-4-nylmethoxy) carbonyl are obtained.] melting point of 72 -73 C (68 C).

  <Desc / Clms Page number 37>

 
 EMI37.1
 1- (phenylmethoxy) [Dissolve phenylmethyl ester of 1- [[(phe-alanyl] -L-proline (2.82 g; 6 mmol) of Example 1 (c) in tetrahydrofuran ( 20 ml) and stir the solution in an ice bath. Add oxalyl chloride (0.63 ml; 7.2 mmol) followed by 4 drops of dimethylformamide. After stirring this reaction mixture in a ice for 20 minutes, then stirred at room temperature for an additional hour.

   The solvents are separated in vacuo, the residue is redissolved in tetrahydrofuran (10 ml) and is returned to an ice bath. With this cold stirring solution, we
 EMI37.2
 add a cold solution of 2-phenyl-4-nylmethoxy) carbonyl] (2.2 gr; 6 mmol) in tetrahydrofuran (14 ml), this addition being followed by the addition of triethylamine (0.85 ml; 6 mmol ). The ice bath is removed and the reaction mixture is stirred at room temperature overnight. The precipitated triethylamine hydrochloride is filtered off and the mother liquor is concentrated under vacuum. It is then redissolved in pyridine (6 ml), 4-dimethylaminopyridine (30 mg) is added, and the solution is stirred at room temperature for 3 hours.

   Acetic acid (6 ml) is added and the reaction mixture is heated at 105 ° C for 45 minutes. It is then evaporated, it is taken up in ethyl acetate, and it is washed

  <Desc / Clms Page number 38>

 with water, dilute hydrochloric acid and aqueous sodium bicarbonate. After evaporation of the solvent, the crude product (3.8 gr) is chromatographed (silica gel; 230 gr) using a mixture of ethyl acetate and hexane (4/3) followed by a mixture of ethyl acetate and hexane (2/1) for the elu
 EMI38.1
 tion, which allows to obtain 1 gr of phenylmethyl ester of 1- [N- [3-methoxy) -carbonyl] amino] -2-oxoheptyl] -N- thoxy) -carbonyl] -L-alanyl] -L -proline. d) 1- [N- [7-amino-3- dihydrochloride; 8oxoheptyl] -L-alanyl] -L-proline.



   Phenylmethyl ester of
 EMI38.2
 lN amino] -2-oxoheptyl] -N- [alanyl] -L-proline (1, 3 gr; 1, 6mmole) in ethanol (75 ml) and aqueous hydrochloric acid (1N; 5 ml) .



  Palladium on carbon catalyst (10%; 450 mg) is added and the mixture is hydrogenated at atmospheric pressure overnight. The catalyst is filtered off and the solution is evaporated in vacuo. The residue is redissolved in water and lyophilized.



  The lyophilization product is triturated with ether so as to obtain 0.6 g of 1-N [7-amino-3- (benzoylamino) -2-oxoheptyl] -L-alanyl] -L-proline hydrochloride;
 EMI38.3
 melting point of 80 -155 C; [a] = D -500 (c = l, l, methanol). Rf 0.09 (silica gel; n-butanol / acetic acid / water; 4/1/1).

  <Desc / Clms Page number 39>

 
 EMI39.1
 Analysis: calculationforCHN0. 5
C 49, 63; H: 7.00; N: 10.52; C1: 13.55 Found: C: 49.63; H: 6.82; N: 10.48; Cl: 13.36.



   Example 3.



  1- [N- [3- (benzoylamino) -2-oxo-heptyl] -L-alanyl] -L-proline monochloride. 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, it is added over a period of 30 minutes sodium hydroxide (2N, 150 ml) and benzoyl chloride (330 mmol; 38.3 ml). 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 filtered to give 68.9 g of N-benzoyl-D, L-norleucine; melting point of 131 -L33 C (1250C). b) 4-butyl-2-phenyl-5 (4H) -oxazolone.



   N-benzoyl-D, L-norleucine (40 g; 170 mmol) is taken up in tetrahydrofuran (300 ml) with 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 filtered off and the tetrahydrofuran is concentrated in vacuo.

   The residue is purified (31.7 gr) on silica in a mixture of hexane and ether

  <Desc / Clms Page number 40>

 (2/1) to give 32.1 g of 4-butyl-2-phenyl-5 (4H) oxazolone. c) 1- [N- [3- (benzoylamino) - 2-oxoheptvl] -N- [(phenylmethoxy) carbonyl] -L-alanvl] -L-proline phenylmethyl ester.



   One cools in an ice bath
 EMI40.1
 solution of 1-methyl) -N-line phenylmethyl (1.441 g; 3 mmol) from Example 1 (c) in tetrahydrofuran (10 ml). While stirring, oxalyl chloride (0.32 ml; 3.7 mmol) is added followed by 4 drops of dimethylformamide. The reaction mixture is stirred in the ice bath for 20 minutes and then at room temperature for 1 hour. Evaporated in vacuo, redissolved in tetrahydrofuran (5 ml) and cooled in an ace bath. A cold solution of 4-butyl-2-phenyl-5 (4H) -oxazolone (0.65 g; 3 mmol) in tetrahydrofuran (5 ml) is added dropwise, this addition being followed by the addition of triethylamine (0.43 ml; 3.1 mmol).

   The reaction mixture is stirred at room temperature overnight.



  After having separated the triethylamine hydrochloride by filtration, the tetrahydrofuran solution is concentrated under vacuum. The residue is redissolved in pyridine (3 ml), 4-dimethylaminopyridine (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. It is then evaporated, it is redissolved in ethyl acetate and

  <Desc / Clms Page number 41>

 it is washed with water, saturated sodium bicarbonate, dilute hydrochloric acid and water.

   After evaporation, the residue is chromatographed on silica gel using the solvent system formed of ethyl acetate and benzene (4/6) to
 EMI41.1
 give 0.8 g of 1- [N- [3- (benzoylamino) -2-oxoheptyl] -N-carbo- [(phenylmethoxy) nyl] -L-alanyl] -L-proline phenylmethyl ester. d) 1- [N- [3enzoylamino) -2-oxoheptyl] -L-alanyl] -L-proline hydrochloride.



   Phenylmethyl ester is dissolved
 EMI41.2
 l-N [- [3-methoxy) carbonyl] -L-alanyl] -L-proline (0.96 g; 1.5 mmol) in ethanol (75 ml). Hydrochloric acid (IN; 2 ml) is added and then 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, the residue is dissolved in water and lyophilized to give
 EMI41.3
 0.62 g of 1- [N- [3-oxoheptyl] -L-alanyl] -L-proline monohydrochloride melting point 860-123 C; [a] -62, (c = 1.05 methanol). Rf 0.57 D f (silica gel; n-butanol / acetic acid / water; 4/1/1).



  Analysis: calculated for C22H31N3O5. HCl. 2H2O:
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
Example 4.
 EMI41.4
 



  1-pyridinvl) butyl dihydrochloride] -L-alanvl] -L-proline. a) 2- ropenoic acid.

  <Desc / Clms Page number 42>

 



   [N- [3- (benzoylamino) -2-oxo-4- (3-Dissolve in acetic acid (24 ml) and in aqueous hydrochloric acid (0.5N; 150ml) of 2- phenyl-4- (3-pyridinylmethylene) -5 (4H) -oxazolone (3 g; 12 mmol) [see Griffith et al., J.



  Org. Chem. Flight. 29, p. 2659]. The reaction mixture is stirred overnight at room temperature.



  It is evaporated and it is again evaporated in absolute ethanol. It is triturated with tetrahydrofuran, it is filtered and the filtered solid is shredded with
 EMI42.1
 absolute ethanol to give 2.8 g of amino acid) of fusion of 215 -216 C (2030C). b) 2- Acid 2- Propenoic acid (14 gr; 46mmol) is dissolved in water (500 ml) and hydrogenated using a palladium on carbon catalyst (10%; 1.8 gr) during the night.



  The catalyst is filtered off, the reaction mixture is evaporated in a small volume (100 ml) and lyophilized to give 13.1 g of product. The lyophilization product is triturated with a mixture of absolute ethanol and ether and filtered to give 12 g of 2- (benzoylamino) -3- (3-pyridinyl) -2-propanoic acid; melting point of 990-1150C. c) 1- [N- [3- (benzoylamino) -2- oxo-4- (3-pyridinyl) butyl] -N - [(phenylmethoxy) carbonyl] - L-alanyl] -L-proline phenylmethyl ester.



   Phenylmethyl ester of 1- [N- (carboxymethyl0-N - [(phenylmethoxy) carbonyl] -L- alanyl] -L-proline (6.2 g; 13.1 mmol) of Example 1 (c is dissolved) ) in tetrahydrofuran (20 ml) and the

  <Desc / Clms Page number 43>

 solution in an ice bath. Oxalyl chloride is added and then 4 drops of dimethylformamide. After stirring this reaction mixture in an ice bath for 20 minutes, it is then stirred at room temperature for an additional hour. The solvents are removed under vacuum and this residue is redissolved in tetrahydrofuran (20 ml).



  2- (Benzoylamino) -3- (3-pyridinyl) -2-propanoic acid (4 g; 13 mmol) is suspended in tetrahydrofuran (45 ml) and while stirring in an ice bath, add triethylamine (1.96 ml; 14 mmol) and dicyclohexylcarbodiimide (2.96 gr; 14 mmol). The reaction mixture is stirred at room temperature overnight. It is then filtered, and the filtrate is evaporated to dryness. This residue is dissolved in tetrahydrofuran (30 ml) and stirred in an ice bath. We add to this solution the
 EMI43.1
 previous solution of 1-carbonyl] -L-alanyl] -L-proline phenylmethyl ester in tetrahydrofuran (20 ml). Triethylamine (1.9 ml; 13.6 mmol) is added and the reaction mixture is stirred at room temperature overnight.

   It is 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 1000C for 45 minutes. It is then evaporated, it is redissolved in ethyl acetate and it is

  <Desc / Clms Page number 44>

 wash with aqueous sodium bicarbonate and water.



  After evaporation, the ethyl acetate extract is chromatographed on silica gel using ethyl acetate for the elution, which allows 3.3 g of phenylmethyl ester to be obtained.
 EMI44.1
 1- [N- [3- [d) 1- [N- [3- (benzoylamino) -2-oxo-4- (3-pyridinyl) butyll-L-alanyll-L-proline hydrochloride.



   Dissolve the phenylmethyl ester product from paragraph (c) (2.6 g; 3.84, mmol) in ethanol (75 ml) and add aqueous hydrochloric acid (1N; 8 ml) followed by a palladium on carbon catalyst (10%; 0.6 gr). After hydrogenation for 16 hours, an additional 0.5 g of the 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 of paragraph (c).



  The hydrogenated material is then chromatographed on LH-20 in water so as to obtain the homogeneous product. An aqueous solution of this material is treated with aqueous hydrochloric acid (1N; 3ml) and the solution is lyophilized to give 1.0g of 1- [N- [3- (benzoylamino) - 2-oxo dihydrochloride -4- (3-pyridinyl) butyl] -L-alanyl] -L-proline; point
 EMI44.2
 120-135 C; [a] 2 55, (c = 1, 1; methaD
50nol). Rf 0.11 (silica gel; n-butanol / acetic acid / water; 4/1/1).

  <Desc / Clms Page number 45>

 
 EMI45.1
 



  Analysis calculated for C: 51, 35; H 5.75; N: 9.98; Cl: 12, 63 Found:: 51, 35; H 5.84: N: 9.96; Cl: 12, 84.



  Example 5.



  1- [N- [3- (4-hydroxy-phenyl) -2-oxobutyl a) 2-P, 5 (4H) -oxazolone monohydrochloride.



   0-Benzyl-L-tyrosine (11.0 g; 40.5 mmol) is taken up in 0.5N sodium hydroxide (81 ml) and water (81 ml) while stirring intensively in an ice bath. To this is added in 5 equal portions in all 52 ml of benzoyl chloride, 45 ml of IN sodium hydroxide and an additional amount of 400 ml of water over a period of 25 minutes. The bath is removed and the reaction is allowed to continue 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 filtered so as to obtain 12.9 g of N-benzoyl-O-benzyl-L-tyrosine;
 EMI45.2
 melting point of 166 -168 C (162 C).



  This N-benzoyl-O-benzyl-L-tyrosine (12.76 g; 35 mmol) is taken up in dry tetrahydrofuran (50 ml) while stirring in an ice bath. To this is added dropwise dicyclohexylcarbidimide (7.7 g; 37.4 mmol) in tetrahydrofuran (18 ml). After 20 minutes, the bath is removed. of ice and the reaction is allowed to continue overnight at room temperature. We separate by

  <Desc / Clms Page number 46>

 filtration the dicyclohexylurea and the filtrate is concentrated to dryness. The crude product is crystallized from a mixture of ether and hexane to give 10.26 g of
 EMI46.1
 2-phenyl-4-oxazolone; melting point of 85 -87 C (830C). b) 1- [N- [3- oxo-4-methoxy) carbonyl] -L-alanyl] -L-proline phenylmethyl ester.



  Dissolving phenylmethyl ester of 1-carboxymethyl) -N- L-alanyl] -L-proline (2.82 g; 6 mmol) from Example 1 (c) in tetrahydrofuran (20 ml) and stir the solution 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, it is then stirred at room temperature for an additional hour. The solvents are separated in vacuo, the residue is redissolved in tetrahydrofuran (10 ml) and cooled in an ice bath. With this cold stirring solution, we
 EMI46.2
 add a cold solution of 2-phenyl-4-methoxy) phenyl] methyl] -5 (2.14 g; 6 mmol) in tetrahydrofuran (40 ml).

   This triethylamine (0.84 ml; 6 mmol) is added and a basic atmosphere is maintained throughout the reaction by adding additional necessary amounts of triethylamine. The reaction mixture is stirred at room temperature overnight.



  It is then filtered, the filtrate is evaporated and redissolved in pyridine (7 ml). We add

  <Desc / Clms Page number 47>

 4-dimethylaminopyridine (30 mg) 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. It is then evaporated, the residue is redissolved in ethyl acetate and washed with saturated sodium bicarbonate and dilute hydrochloric acid.

   The extract is evaporated with neutral ethyl acetate and chromatographed on silica gel (300 g) using the solvent system formed from ethyl acetate and benzene (6.5 / 3.5) so as to obtain 2.7 gr of phenylmethyl ester of 1- [N-
 EMI47.1
 [3- butyl] -N- c) 1- [N- [3- hvdroxyphenyl) hydrochloride
The ester product from paragraph (b) (1.8 g; 2.26 mmol) is dissolved in ethanol (150 ml) containing aqueous hydrochloric acid (1N; 3.5 ml) . Palladium on carbon catalyst (10%; 500 mg) is added and the solution is stirred under a hydrogen atmosphere overnight.

   It is evaporated, dissolved in water and lyophilized to
 EMI47.2
 give 1- [N- [3- 4-4-hydroxyphenyl-2-oxobutyl] -L-alanyl] -L-proline monohydrochloride; melting point of 118 -152 C; [o = 63, 10 (c = 1.07, D methanol). Rf 0.47 (silica gel; n-butanol / acetic acid / water; 4/1/1).



  Analysis: calculated for C25H29N306. HCl. H20:
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.

  <Desc / Clms Page number 48>

 



   Examples 6 to 62.



   Following the procedure of the examples. 1 to 3 and 5, the peptide ester represented in column I is treated to give the carboxymethyl peptide ester represented in column II. The conversion to its acid chloride and the subsequent reaction with the oxazolone from column III gives the N-protected ester product from column IV. Separation of the protecting group from N and from the ester group gives the final product of column V, in which R6 represents hydrogen.



   Collar. I
 EMI48.1
 Collar. II
 EMI48.2
 Collar. III
 EMI48.3
 

  <Desc / Clms Page number 49>

 
 EMI49.1
 Collar. IV
 EMI49.2
 
 EMI49.3
 Collar.
 EMI49.4
 

  <Desc / Clms Page number 50>

 
 EMI50.1
 



  EXemP1e R.-R Example / - \ "'&commat; -' r /. O) Cl 'H OC) 7 H (L) 7 H3C -' &commat; -CH 2-H - N-, COOCHH

  <Desc / Clms Page number 51>

 
 EMI51.1
 Example R3 (oY-c., - - COOCH &commat; -CH2- - -n "H / - \ / - \ 9 ClC- (0) -CH-C-) / --k" 3 N3 1 H N3 10 (0) -2-) -2- H,), - t '10 H

  <Desc / Clms Page number 52>

 
 EMI52.1
 Example 1 Ll &commat; CH2- H 12 c-, - -r (L) '- / H 2 H F 1rj 2 s 1 (L) H H

  <Desc / Clms Page number 53>

 
 EMI53.1
 Example R1 - 0 0 'OCH2- 0 -' H2H U K3) N N (L) H 2 0 0 15 ')) - X 15 H 16 16; J.



  H

  <Desc / Clms Page number 54>

 
 EMI54.1
 X Example S co- ' <0) - <= '', - <F H 0 il NH-C-CH &commat; -HL "L" S H - L-COOCH- (L) H 19 (()) - "2- 0 1 COOCH H

  <Desc / Clms Page number 55>

 
 EMI55.1
 Example 3 0 0 0 20 H COH C-. N 0-CH 2- 20 0 / o) -OH2C-'o) '- 2-r 21 H) H.



  21 CH2 CH 0-c 0-00 Co 22 S

  <Desc / Clms Page number 56>

 
 EMI56.1
 R 3 Example 1 R2 - b OCN (CH) H 23 H, C, - M-oeCH, (L) VJL "2's 3 -COOCH- H" H 0-Cil H 25 3-N COOCH 2

  <Desc / Clms Page number 57>

 
 EMI57.1
 Example R R2 26? ) - Y''H (L) 3 3 0 3 i / \ 7 "-N-- J o 0 27 H02-Y2COOCHO) H H H H (o) - &commat; -" = ", ' <-r 29 H

  <Desc / Clms Page number 58>

 
 EMI58.1
 Example ¯ 30 31 HH - H 32 HC-ft'L 2 '' ---) - \ H 32 HC s Ca 2-'q7COOCH 5 g - \ / 2-N --- L-coeCHH (L) 2 0H "'33 3 H

  <Desc / Clms Page number 59>

 
 EMI59.1
 Example - - \ &commat; H 34 Hic-. (0) '"2'OrC H H C- &commat; -CH2- C9l-H- ct? \ 3S H" - -.



  C-HN- H 36, - <H, - 2-r 2 it

  <Desc / Clms Page number 60>

 
 EMI60.1
 Example dz 2 3 os 37 Il Il 37 M --- COOCH- H 0 &commat; -H, -H,) -CH, - 38 H -P- .., (L) H 39 H co-2 2Cs 00- (CH22H H2CO HH

  <Desc / Clms Page number 61>

 
 EMI61.1
 Example Rl R2 "frr" 'L' C) H 0 40 N 40 1 41 Jr Lr 2 0 42 C-S- -N - p-coecH-Q) 'H

  <Desc / Clms Page number 62>

 
 EMI62.1
 R3 Example UN 43 2 3 "Ù) / - / 2 - 'cooeHD) O.) 3 45 H, -) - 3) coC-C", -) .-.



  3 2 s 45 H3c-17 H2c- -N COOCH H H H

  <Desc / Clms Page number 63>

 
 EMI63.1
 Example Rl - - - - 46 HC- '"2'-NH-CH-COOCH) 47 FC-rQ'" 2 -0) 3 CH3 '"2'" '-NH-CH-COOCH 1 (L) 2 1H2 1 2 CH) 49 H) -NH-CH-COOCH- / 0) 1 (L) 2 cl CH ocii OCH

  <Desc / Clms Page number 64>

 
 EMI64.1
 Example R - - - - 50 HC-0) '-)' C "'- NH-CH-COOCH- / 0) 3 1 CH 2 OCH, -) 51 H C- (0) -2 - MH-CH -COOCH 3 Cil 1 0 H 52.



  .



  '"nrij 1 r <

  <Desc / Clms Page number 65>

 
 EMI65.1
 Example R. R2 R3 2 3 53 3 1 (L) 2 (CH2) -NHCOCH- 2 o 54 -NH-CH-COOCH-5) 3 S -S-CH-) 55 H - "-" H (CH ) 3NHC.



  NHNO 56 H, 0) -) -2--2- &commat; (112) 2 0

  <Desc / Clms Page number 66>

 
 EMI66.1
 Example R2 \ &commat; CH2- 57 JS 58 3 "Il --N --- C-0-CH-OCC in L) 1:,) SQ qJo 59 H, 23 ------ 2" 5 NNH CH )

  <Desc / Clms Page number 67>

 
 EMI67.1
 R) Example 1 0 0 60 FJ-- - Il '- S (L) H 61 (0) -' H2-) 0 3 H 0 62 H- - 2-r f) 3 C-O Il (L) H

  <Desc / Clms Page number 68>

 
The protection groups R in examples 20, 36 to 39 and 41, the protection groups
R3 in Examples 43 and 44, and protecting groups R5 in Examples 49.50 and 52-55 are separated as the last step in the synthesis.



  The R6 ester groups shown in Examples 57 to 62 are not separated.



   Example 63.
 EMI68.1
 



  1- [N- [oxo-4-phenvlbutyl] -L-alanyl] -L-proline hydrochloride. a) Hydrogen bromide of (S) - 3-amino-1-chloro-4 phenyl-2-butanone.



   Phenylmethyl ester of (S) - [3-chloro-2-oxo-1- (phenylmethyl) propyl] carbamic acid (51.4 g) is dissolved in a mixture of acetic acid (252 ml) and hydrogen bromide in acetic acid (3.45 N; 348 ml) and the mixture thus obtained is kept at room temperature for 1.5 hours. The reaction mixture is then concentrated in vacuo and precipitated with ether so as to obtain 36.6 g of hydrogen bromide of (S) - 3-amino-1-chloro-4phenyl-2-butanone; melting point of 1770-1790C (1750C). b) (S) -N- [3-chloro-2-oxo-1- (phenylmethyl) propyl] benzamide.



   (S) -3-amino-1-chloro-4-phenyl-2-butanone hydrogen bromide (36.3 g; 130.3 mmol) is suspended in 520 ml of dry tetrahydrofuran and 18.2 ml of triethylamine (130.3 mmol) with stirring for 10 minutes. The mixture is placed in an ice bath and 15.2 ml of benzoyl chloride are added.

  <Desc / Clms Page number 69>

 followed by 10.95 g of sodium bicarbonate. After 5 minutes, the ice bath is removed and the reaction mixture is kept at room temperature for 1.5 hours. The reaction mixture is then concentrated in vacuo and the residue is taken up in 1 liter of aqueous methanol (10% water).

   The precipitate is collected, filtered and washed with metal.
 EMI69.1
 thanol so as to obtain 25.3 g of (S) -N- 2-oxo-1- melting point of 170 -172 C (decomposition) (1600C); [at] ; (c = 1, 7; D c) 1,1-Dimethylethyl ester of 1- [N- [amino) [3-chloro-L-alanyl-L-proline 1,1-dimethylethyl ester is combined ( 2.42 gr; 10 mmol), sodium bicarbonate (840 mg) and (S) -N- [3-chloro-2-oxo l- (phenylmethyl) propyl] benzamide (3.01 gr) in
50 ml of dimethylformamide under an atmosphere of argon at room temperature with stirring overnight.

   The reaction mixture is then concentrated in vacuo to half its initial volume, the residue is taken up in ethyl acetate and washed with saturated sodium bicarbonate so as to obtain 2.25 g of gross product. This material is taken up in a mixture of ethyl acetate and methanol (95/5) and applied to a column of silica gel (135 g), which is eluted with a mixture of acetate d ethyl and methanol (95J5) so as to obtain 860 mg of 1,1-dimethylethyl ester
1- [N - [(S) -3- (benzoylamino) -2-oxo-4-phenylbutyl] -L- alanyl] -L-proline.

  <Desc / Clms Page number 70>

 
 EMI70.1
 d} 1-lN = j oxo-4-phenylbutyl] -L-alanyl] -L-proline monohydrochloride.



  1,1-dimethylethyl ester of l-oxo-4-phenylbutyl] -L-alanyl] -L-proline (740 mg; 1.46 mmol) is dissolved in a solution of hydrogen chloride in acetic acid (1.5N; 10 ml) and the reaction mixture is kept at room temperature for 30 minutes. It is then concentrated, it is taken up in water, it is filtered and it is lyophilized to
 EMI70.2
 obtain 600 mg of 1- (benzoylamino) -2-oxo-4-phenylbutyl] -L-alanyl] -L-proline monohydrochloride; melting point of 83 -163 C; D (c = 1.04; methanol) 'R 0.6 (silica gel; butanol / acetic acid / water; 4/1/1).



  Analysis: calculated for C25H29N3O5.HCl.1.65 H2O:
C: 59.99; H: 6.48; N: 8.12; Cl: 6.85 Found C: 57.99; H: 6.39; N: 8.09; Cl: 6.95.



   Example 64
 EMI70.3
 4-phenylbutyl] -L-lysyl] -L-proline dihydrochloride. a) 1,1-Dimethylethyl ester of 1- - (S) -1- [N2¯ [3- (benzoylamino) -2-oxo-ethoxy) carbonyl] -N2 - [(phenylmethoxy) carbonyl] -L-lysyl] -
L-proline.



   N- [(1,1-dimethylethoxy) carbonyl) -N2 - [(phenylmethoxy) carbonyl] -L-lysine (9.51 g) and hydroxybenzotriazole (3.825 g) are taken up in 25 ml of dimethylformamide with stirring in an ice bath under an argon atmosphere.



  To this is added 1,1-dimethylethyl ester of L-proline (4.49 g) followed by N, N'-diisopropylethylamine (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.

  <Desc / Clms Page number 71>

 



  Dimethylformamide is separated under vacuum. The residue is taken up in ethyl acetate and the dicyclohexylurea is filtered off. The filtrate is washed until neutral with 10% potassium bisulfate and saturated sodium bicarbonate. The crude product (13.26 gr) is purified on a column of silica gel, eluting with a mixture of ethyl acetate and hexane (2/1) so as to obtain 13.0 gr of ester l- [N - [(1,1-dimethylethoxy) - l-dimethylethyl -
 EMI71.1
 0 carbonyl] -N- proline. b) Ester 1,1-dimethylethyl of (S) -1- [N2- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N6- [(1,1-dimethylethoxy) - carbonyl] -L- lysyl] -L-proline.



   The ester product from paragraph (a) is reduced in ethanol with palladium on carbon catalyst (10%) so as to obtain 3.99 g of 1, 1-dimethylethyl ester of 1- [N6¯ [1,1dimethylethoxy) carbonyl] -L-lysyl] -L-proline. This material is taken up in 40 ml of dimethylformamide and treated with (S) -N- [3-chloro-2-oxo-1- (phenylmethyl) propyl] benzamide (3.01 gr) coming from the example 63 (b) and with sodium bicarbonate (840 mg).



  After stirring for 18 hours at room temperature, the reaction mixture is concentrated in vacuo, taken up in ethyl acetate and washed with saturated sodium bicarbonate. The crude product (7.0 g) is purified on a column of
 EMI71.2
 silica, eluting with a mixture of ethyl acetate and methanol (1%) to give 1.7 g of 1,1-dimethylethyl ester of (S) -1- [N- oxo-4-phenylbutyU- NOT-

  <Desc / Clms Page number 72>

 
 EMI72.1
 L-lysyl] -L-proline. o c) [3- (Benzoylamino) -2-oxc-4-phenylbutyl] -L-lysyl] -L-proline dihydrochloride.



   The ester product of paragraph (b) (1.6 g) is treated for 30 minutes at room temperature with 20 ml of 1.5N hydrochloric acid and acetic acid, concentrated to dryness, and triturated with ether to obtain a solid to give 1.37 g of crude product. This material is taken up in water, filtered through a millipore filter, and lyophilized to give 1.28 g of product. Further purification is carried out on an LH-20 column in water to give 740 mg of
 EMI72.2
 - 4-phenylbutyl] -L-lysyl] -L-proline dihydrochloride; melting point of 120-180 C; [at] ; (c = 1.05; methanol).



  D 0.61 (traces at 0.9) (silica gel; chloroform / methanol / acetic acid; 60.38.2%).



  Analysis: for CHN0.2HC1.3H C: 52, 98; H: 6.88; N: 8, 83: 11, 17 Found: C: 52.98; H: 6.93; N: 8.66; Cl; 11, 31.



   Example 65.
 EMI72.3
 



  N-oxo-4-phenylbutyl] -L-alanyl] -N-Cyclohexylqlycine monohydrochloride. a) 1,1-Dimethylethyl ester of N-cyclohexylqlycine.



   [N- [[(S) -3- (benzoylamino) -2-Cyclohexylamine (70.35 ml) and sodium bicarbonate (12.9 g) are suspended with stirring in 200 ml of absolute ethanol while stirring in an ice bath. To this is added dropwise 1,1-dimethylethyl ester of bromoacetic acid (20.78 ml). We remove the ice bath, after

  <Desc / Clms Page number 73>

 24 hours at room temperature, the reaction mixture is concentrated to dryness, it is taken up in chloroform and washed with water.

   The crude product (42 g) is chromatographed on silica gel, eluting with a mixture of ethyl acetate and hexane (2/1) to give 27.4 g of 1,1-dimethylethyl ester of N-cyclohexylglycine. b) 1,1-Dimethylethyl ester of N-cyclohexyl-N- [N- [(Dhenylmethoxv) carbonvl] -L-alanyl] glycine.



   N- [(phenylmethoxy) carbonyll- is stirred
 EMI73.1
 L-alanime (4.46 g), N-cyclohexylglycine (4.26 g), hydroxybenzo-triazole (3.06 g), dicyclohexylcarbodiimide (4.12 g) and triethylamine (2, 8 ml) in 40 ml of dimethylformamide at room temperature for 20 hours.



  The reaction mixture is then concentrated in vacuo, taken up in ethyl acetate, the dicyclohexylurea is filtered off, and the filtrate is washed until neutral with 10% potassium bisulfate and sodium bicarbonate. sodium saturated so as to obtain 5.9 gr of crude product. Crystallization from a mixture of ether and hexane gives 3.97 g of 1,1-dimethylethyl ester of N-cyclohexyl- N- [N- [(phenylmethoxy) carbonyl] -L-alany. l] glycine; melting point of 104 -105 C. c) 1,1-Dimethylethyl ester of N- (L-alanvl) -N-cyclohexylqlycine.



   The ester product from paragraph (b) (3.9 g) is taken up in methanol with palladium on carbon catalyst (10%; 700 mg) and stirred under an atmosphere of hydrogen for 4 hours. We filter the

  <Desc / Clms Page number 74>

 reaction mixture and concentrated to dryness so as to obtain 2.65 g of 1,1-dimethyl ethyl ester of crude N- (L-alanyl) -N-cyclohexylglycine. d) 1,1-Dimethylethyl ester of N- [N - [[(S) -3- (benzoyl-amino) -2-oxo-4-phnéylbutyl] -L-alanyl] -N-cyclohexyl-qlycine.



   The crude ester product of paragraph (c) (2.6 gr), sodium bicarbonate (764 mg) is stirred for 25 hours in 25 ml of dimethylformamide.
 EMI74.1
 and (S) -N- benzamide (2.75 g) from Example 63 (b).



  The reaction mixture is concentrated to dryness, it is taken up in ethyl acetate and washed with saturated sodium carbonate so as to obtain 4.7 g of crude product. Purification on a column of silica gel, eluting with a mixture of ethyl acetate and methanol (99/1), gives 1.5 g of 1,1-dimethyl ethyl ester of N- [N- [[( S) -3- (benzoyl-
 EMI74.2
 amino) glycine. e) Oxo-4-phenylbutyl] -L-alanyl] -N-cyclohexYlqlycine monochlorhydrate.



   -2-oxo-4-phenylbutyl] -L-alanyl] -N-cyclohexyl-The ester product of paragraph (d) (500 mg) is treated for 30 minutes with 5 ml of a mixture of hydrochloric acid 1, 5N and acetic acid and then concentrated to dryness at room temperature.



  The crude product is taken up in methanol and purified on an LH-20 column to give 412 mg of product. This product is made semi-crystalline in a mixture of acetonitrile and ether to give N- [N - [[(S) -3- (benzoylamino) -2-oxo-) monohydrochloride.

  <Desc / Clms Page number 75>

 
 EMI75.1
 4-phenylbutyl] -L-alanyl] -N-cyclohexylglycine; melting point of 154 -157 C (1320C); [a] D '"" D (c = 1.35; methanol). Rf 0.60 (small impurities at 0.92) (silica gel; chloroform / methanol / concentrated ammonia; 30/10/2).
 EMI75.2
 Analysis: for 0 D 3 D 3
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- [N - [[(S) -3- (benzoylamino) -2-oxo-4-phenylbutyl] -L-alanyl] -N-phenylglycine hydrochloride. a) Ester l, l-diméthvléthylique of N-phenylqlycine.



   A solution of triethylamine (11.25 gr; 0.11 mole) and aniline (9.3 gr; 0.10 mole) in ether (100 ml) is cooled to 00 in an ice bath. ) under an argon atmosphere. To this is added 1,1-dimethylethyl ester of bromoacetic acid (18 g; 0.093 mole) over a period of 30 minutes; The resulting mixture is stirred at 00 for one hour, then heated to room temperature and stirred overnight.



  The solution is filtered, rinsed with ether and the filtrate is concentrated to give 6.9 g of a yellow oil. Rf 0.6, 0.7 (silica gel; ethyl acetate). Chromatography on LPS-1 using a mixture of hexane and ethyl acetate (7/3) as eluent gives 2.3 g of 1,1-dimethylethyl ester of N-phenylglycine in the form of a pale yellow liquid. Rf 0.7 (silica gel; ethyl acetate). b) 1,1-Dimethylethyl ester of N-phenyl-N- [N- [(phenylmethoxy) carbonyl] -L-alanvl] -glycine.

  <Desc / Clms Page number 76>

 



   A solution of N- [(phenylmethoxy) carbonyl] - L-alanine (2.8 gr; 12.7 mmol) in dry tetrahydrofuran (50 ml) under argon is cooled in a dry ice and ethanol bath. To this, N-methylmorpholine (1.28 g; 12.7 mmol) and isobutyl chloroformate (1.73 gr; 12.7 mmol) are added.
 EMI76.1
 



  After 20 minutes, 1,1-dimethyl-ethyl ester of N-phenylglycine (3.7 g; 12.7 mmol) is added.



  The resulting mixture is stirred at -20 ° C for one hour, and 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,
 EMI76.2
 dried (MgSO 4) and concentrated. Chromatography on LPS-1, eluting with a gradient of ethyl acetate and hexane f3 / 1 gives 4.0 g of N-phenyl-N-methoxy) carbonyl] -L 1,1-dimethylethyl ester. -alanyl] in the form of a clear oil. Rf 0.4, (silica gel; ethyl acetate). c) 1,1-dimethylethyl ester of N- (L-alanyl) -N-phenylglycine.



   A solution of the ester product of paragraph (b) (4.0 g; 9.7 mmol) in methanol (125 ml) and palladium catalyst (10%) is stirred under a circulation of hydrogen for 18 hours. on carbon. The solution is filtered, concentrated, dissolved in ethyl acetate and extracted with 1N hydrochloric acid. The aqueous layer is treated with sodium bicarbonate until it becomes basic and extracted with

  <Desc / Clms Page number 77>

 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 solid White.



   Rf 0.52; small spots at 0.64 (silica gel; ethyl acetate / methanol; 1/1). d) 1,1-Dimethylethyl ester of N- [N - [[(S) -3- (benzoyl-amino) -2-oxo-4-phenylbutyl] -L-alanyl] -N-phenylglycine.



   Sodium bicarbonate (0.38 g; 4.7 mmol) and sodium iodide (0.7 g; 4.7 mmol) are added.
 EMI77.1
 to a solution of (S) -N-methyl) propyl] (1.4 g; 4.7 mmol) from Example 63 (b) and of 1,1-dimethylethyl ester of N- (L-alanyl ) -N-phenylglycine (1.3 gr; 4.7 mmol) in dry dimethylformamide with stirring. 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 (MgSO.) And concentrated to a yellow oil.

   Chromatography
 EMI77.2
 on LPS-1, eluting with an ethyl acetate / hexane gradient (1/1) until ethyl acetate gives 1.8 g of N, 3-, N-3- [N- [[(S) -phenylglycine. Rf 0.34 (silica gel; ethyl acetate). e) N- [N- [[(S) -3- (benzoylamino) -2-oxo-4-phenylbutyl] -L-alanyl] -N-phenylqlycine monochloride.



   A solution of the ester product of paragraph (d) (1.6 g; 2.9 mmol) in a mixture is stirred at room temperature for 30 minutes.

  <Desc / Clms Page number 78>

 hydrochloric acid and acetic acid (1.77N; 17.0 ml). The solution is concentrated and the residue is triturated with ether so as to obtain a pale yellow solid. Recrystallization from a mixture of methanol and ether gives 0.84 g of mono-
 EMI78.1
 N- [N - [[phenylbutyl] -L-alanyl] -N-phenylglycine hydrochloride in the form of a white crystalline solid; melting point of 147 -159 C (decomposition). Rf 0.8 (silica gel; butanol / acetic acid / water; 1/1/1). [a] = -12.7 (c = 1.5; methanol).
 EMI78.2
 



  Analysis calculated for 0.65 H20 0 Z *? J 3 Z
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.
 EMI78.3
 



  (S) -l- 4-phenylbutyl] -N-methyl-L-alanyl] -L-proline monohydrochloride. a) N- [N- [[3- (benzoylamino) -2-oxo-L-alanine (89.1 g; 1 mole) is dissolved in 2N sodium hydroxide (500 ml) and cooled to 00. To this, benzyl chloroformate (204.5 g; 1.2 mole) and 4N sodium hydroxide (250 ml) are added simultaneously dropwise over a period of one hour. The reaction mixture is stirred overnight (00 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 (NaSO), concentrated on a rotary evaporator and the solid residue is triturated

  <Desc / Clms Page number 79>

 with petroleum ether so as to obtain 194.0 g of N- [(phenylmethoxy) carbonyl] -L-alanine in the form of a white solid. b) N-methyl-N- [(phenylmethoxy) carbonyl] -L-alanine.



   Add a dispersion of sodium hydride (14.25 g; 0.3 mole) carefully
 EMI79.1
 with gentle stirring to a cold solution (00) of N- (22.3 g; [(phenylmethoxy) carbonyl) -L-alanine0.1 mole) and methyl iodide (50 ml; 0.8 mole) in tetrahydrofuran (250 ml). The suspension is stirred overnight under a nitrogen atmosphere (00 to room temperature), poured slowly into saturated sodium bicarbonate (200 ml), diluted with ethyl acetate ( 300 ml) and the layers are separated. The organic layer is extracted once again 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 (Na 2 SO 4) and concentrated in vacuo to a dark oily residue (23.0 g). This crude acid is treated with dicyclohexylamine (20 ml) in ether (150 ml). The resulting crude dicyclohexylamine salt is collected (37.0 gr), recrystallized from a mixture of chloroform and ether (35.0 gr) and converted again to the acid by partitioning between 1N hydrochloric acid and ethyl acetate, which gives a pale yellow oil, which is crystallized by abandonment (17.4 gr).



   Recrystallization (11.2 g) from a mixture of ethyl acetate and petroleum ether gives 4.0 g of

  <Desc / Clms Page number 80>

 N-methyl-N - [(phenylmethoxy) carbonyl] -L-alanine in the form of a white crystalline product; point of
 EMI80.1
 fusion of 650-66.5 C; ] 5 = -31, 10 (c = 2, D acetic acid). c) Ester l, l-dimethylethyl of l-methoxv) carbonvl] -L-abnyl] -L-proline.



   [N-methyl-N- [(phenyl-L-proline 1,1-dimethylethyl ester (3.42 g; 20 mm) is added, hydroxybenzotriazole hydrate (3.06 g; 20 mmol) and dicyclohexylcarbodiimide (4.12 gr; 20 mmol) to a solution of N-methyl-n - [(phenylmethoxy) carbonyl] -Lalanine (4.74 gr; 20 mmol) in distilled tetrahydrofuran (50 ml). 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 (2 x), 10% potassium bisulfate (2 x) and water (2 x ), it is dried (Na2SO4) and concentrated to
 EMI80.2
 give 6.4 g of 1,1-dimethylethyl ester of 1- [N-methyl-N- in the form of an oily residue. d) Ester 1, l-dimethylethyl of l- (N-methyl-L-alanyl) - L-proline.



   A mixture of the ester product of paragraph (c) (6.4 gr; 16.4 mmol) and 0.8 gr of palladium catalyst (10%) on carbon in ethanol (95%; 150 ml ) is hydrogenated at atmospheric pressure overnight. The catalysts are separated by filtration and the filtrate is evaporated. The resulting oily residue solidifies on drying in a high vacuum to a residue

  <Desc / Clms Page number 81>

 oily solid (3.8 gr).

   Trituration with ether gives 1.5 g of l- (N-methyl-L-alanyl) L-proline 1,1-dimethylethyl ester monohydrochloride
 EMI81.1
 as a white solid; Rf 0.44 (silica gel; (20%) chloroform methanol). [a] = -102, (c = 2;
The ether filtrate gives 2.3 g of 1,1-dimethylethyl ester of 1- (N-methyl-L-alanyl) -L-proline in the form of an oil; Rf of 0, (gel
 EMI81.2
 silica; methanol (20%) chloroform). 5 = = -101, D (c = 2; acetic acid). e) Ester l, l-. of (S) -1- (benzovlamino) -2-oxo-4-phenylbutyl] -N-methyl-L-alanvl] - [a] L-proline.



   The mixture is stirred at ambient temperature overnight under a. nitrogen atmosphere a reaction mixture of l- (N-methyl-L-alanyl) -L-
 EMI81.3
 proline (2.1 g; 8.19 mmol), of (S) -N- 1- (2.46 g; 8.19 mmol) of Example 63 (b), excess of sodium carbonate and sodium iodide (1.22 g; 8.8 mmol) in dimethylformamide (15 ml). The reaction mixture is concentrated, the residue is partitioned between water and ethyl acetate, the layers are separated, and the aqueous layer is extracted again with ethyl acetate. The combined organic extracts are washed with saturated sodium bicarbonate, and water and dried (Na2SO4) and concentrated in an oily residue (3.5 g).

   Rapid chromatography (200 g of silica gel; 20% methanol / ethyl acetate) gives 2.8 g of 1,1-dimethylethyl ester of (S) -l- [N - [[3- (benzoylamino ) -2-oxo-4-phenylbutyl] -N-methyl-L-

  <Desc / Clms Page number 82>

 
 EMI82.1
 alanyl] -L-proline in the form of a yellow oil. f) (S) oxo-4-phenylbutyl] -N-methyl-L-alanyl] -L-proline hydrochloride.



   The ester product of paragraph (e) (1.4 g; 2.7 mmol) is treated with a mixture of 2N hydrochloric acid and acetic acid (20 ml). After stirring for 2 hours at room temperature, the reaction mixture is concentrated under reduced pressure and the resulting oily residue is triturated with ether (4 times) to give 1.05 g of monohydrochloride
 EMI82.2
 (S) methyl-L-alanyl] -L-proline as a slightly dirty white solid; melting point of 1250-1350; Rf 0.24 (silica gel; n-butanol / acetic acid / water; 4/1/1). [a] -870 (c = 1; methanol).



  D Analysis calculated for 0, 7
C: 60.68; H: 6.41; N: 8.16; Cl: 6.88 Found: C: 60.68; H: 6.36; N: 7.95; Cl: 6, 48.



   Example 68
 EMI82.3
 (S) phenylbutyl] -4-phenylqlycYl] -L-proline hydrochloride (20/3). a) 1,1-Dimethylethyl ester of l- (bromoacetyl) -Lproline.



   Diisopropylethylamine (38.3 ml; 0.22 mole) and bromoacetyl chloride (16.5 ml; 0.2 mole) are added dropwise over a period of 20 minutes while maintaining the temperature between - 10 and -5 C to a cooled solution (-10 C) of 1,1-dimethylethyl ester of L-proline (34.2 g; 0.2 mole) in methylene chloride (250 ml). The dark-colored reaction mixture is stirred overnight

  <Desc / Clms Page number 83>

 (from −10 ° C. to room temperature) and it is concentrated under reduced pressure. The oily residue is redissolved in ethyl acetate, washed with saturated sodium bicarbonate (2 times), 10% potassium bisulfate (2 times) and water (2 times), dried (Na2SO4) and concentrated to a dark oily residue (28.0 gr).

   Chromatography
 EMI83.1
 rapid silica gel (LPS-1; 10% ethyl acetate / methylene chloride) gives 11.0 g of 1,1-dimethylethyl ester of 1- (bromoacetyl) -L-proline in the form of an oil pale yellow. b) Ester 1, l-dimethylethyl of 1- (N-phenylqlvcyl) -L-, proline.



   Aniline (1.5 g; 15 mmol) is added to a solution of 1,1-dimethylethyl ester of 1- (bromo-acetyl) -L-proline (2.1 gr; 7.5 mmol) in distilled tetrahydrofuran (40 ml) and the reaction mixture was stirred overnight under nitrogen. The reaction mixture is diluted with ethyl acetate (200 ml), washed with saturated sodium bicarbonate (2 times 50 ml) and water (2 times), dried (Na 2 SO 4 ) and concentrated in vacuo to an oily residue of dark color (4.0 gr).

   Rapid chromatography (silica gel LPS-1; 10% ethyl acetate / methylene chloride) gives 2.2 g of 1,- (N-phenylglycyl) -L-proline 1,1-methylethyl ester under the form of a dark oil which solidifies on drying in a high vacuum. c) 1,1dimethylethyl ester of (S) -l- [N- [3- enYl) - amino) -2-oxo-4-phenylbutyl] -N-phenylglycyl] -L-proline.



   A mixture of reagents is stirred at room temperature under a nitrogen atmosphere overnight.

  <Desc / Clms Page number 84>

 tion of l- (N-phenylglycyl) -L-proline 1,1-dimethylethyl ester (1.6 g; 3.5 mmol), of (S) -N- [3-chloro-2-oxo -l- (phenylmethyl) propyl] benzamide (1.06 g; 3.5 mmol) of Example 63 (b), an excess of sodium bicarbonate 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 times 50 ml).

   The combined ethyl acetate extracts are washed with saturated sodium bicarbonate (2 times) and water (3 times), dried (Na-SO.) And concentrated under reduced pressure to to obtain a dark oily residue (2.0 gr). Rapid chromatography (silica gel LPS-1; 5% ethyl acetate / methylene chloride up to 15% ethyl acetate / methylene chloride) gives 0.3 g of 1,1-dimethylethyl ester. -
 EMI84.1
 than (S) N-phenylglycyl] -L-proline in the form of a pale yellow foam. d) (S) -l- hydrochloride. [N- -2, -oxo-4phenylbutyl] -N-phenylqlycvl] -L-proline. (20/3) -1- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -The ester product of paragraph (c) is treated (0.28 gr; 0.49 mmol) with a mixture of 2N hydrochloric acid and acetic acid.

   After stirring for 1 hour at room temperature, the reaction mixture is concentrated under reduced pressure and the resulting oily residue is triturated with ether (4 times) to give 0.14 g of hydrochloride.
 EMI84.2
 (S) -l- phenylglycyl] -L-pr61ine (20/3) in the form of a

  <Desc / Clms Page number 85>

 
 EMI85.1
 slightly dirty white solid; melting point of 110-140 C; from 0.76 (small spots 53) a (silica gel; n-butanol / acetic acid / water; 3/1/1.



  Analysis calculated for C30H31N30. HC1. 0.5 H20:
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.



   Following the method of Examples 63 to 68, but using the ketone shown in column I, the acid chloride shown in column II and the peptide ester shown in column III, the ester product is obtained represented in the column
 EMI85.2
 IV. Separation of the ester group or any other protecting group gives the corresponding end product in the form of acid.

  <Desc / Clms Page number 86>

 
 EMI86.1
 Collar.
 EMI86.2
 

  <Desc / Clms Page number 87>

 
 EMI87.1
 



  Example R3 R2 Rl X R s X S H (iO> p-2H Hc- 71) -2 - <) - "- N - OOC c- (L) Il zo (CH H C - N 72 J 22-" 3'COOC us II H 72

  <Desc / Clms Page number 88>

 
 EMI88.1
 Example'R'R - 7 -N --- f-COOC 3 H 74 HN CH 2 s H s 2 HI CH- &commat;) H '"H 75 HC- C) - * -N-CH-COOC) H - 3 2 3 3 &commat; 2 33

  <Desc / Clms Page number 89>

 
 EMI89.1
 Example R 0 o 76 HCOCHN) - (CH '' -M ---- COOC H H S S (o) - -H.



  --C-HN- -COOC / O.



  2 s 78 "f" 3 C-UN- 78 ON-HN

  <Desc / Clms Page number 90>

 
 EMI90.1
 Example L r - "- - c-rY - - <r3 1.



  H 8 ' <-2 "- - s H -H -COOC H H 81 - Qr I H C CH 2 2 82 H poch P" 2

  <Desc / Clms Page number 91>

 
 EMI91.1
 R3 R2 Example 83 IF ÎJ-H CH- 1 CH.



  .



  OCH2 84 n-) - "3 - NH-CH-COOC) H- 2" r ", CH 0. CH3 S5 / YcH- / HC - NH-CH-COOC 3 (Cil) - 'NHCOCH BS 0

  <Desc / Clms Page number 92>

 
 EMI92.1
 Example R R2 R X R - - - 3 1 2 4-3 i (L) (CH) -C-NH 22 2 0 i S 87 2 no q (101, H 0 0 <0) -CH, - - "'" --- C-0-CH-O-C-C' 88 ss H CH (CH3) 2

  <Desc / Clms Page number 93>

 
 EMI93.1
 Example R 2 1 1 1 0 0 H c- 89 3 89) (CH) \ / --- L-C-O-CH-O-C-C) HN H 0 &commat; - O-H -O-fH-O - C2H5 90 3 HH CH3

  <Desc / Clms Page number 94>

 
The protection groups R-in examples 74, 76 and 77, the protection groups R3 in examples 78 and 80 and the protection groups R5 in examples 81 and 83 to 85 are separated as the last step in the synthesis. . The
 EMI94.1
 ester groups R6 in examples 87 to b 90 are not separated.



  Example 91.



  (-) - 1-4-phenylbutyl] -N-methyl monohydrochloride a) [3-Oxo-4-phenylbutyl] methylcarbamic acid phenylmethyl ester.



  N-methyl-N-carbonyllglycine (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 followed by 2 drops of dimethylformamide. After stirring for 30 minutes in the ice bath, the mixture is then stirred at room temperature for 1 hour. To this is added 0.25 ml of oxalyl chloride. The mixture is evaporated, redissolved in 15 ml of tetrahydrofuran, and stirred in an ice bath. To the above solution is added while stirring in the ice bath a solution of 2-phenyl-4- (phenylmethyl) -5 (4H) -oxazolone (3.1 g; 12.4 mmol) dissolved in 15 ml of tetrahydrofuran. 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,

  <Desc / Clms Page number 95>

 then redissolved in pyridine (5 ml) and this 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 a mixture of aqueous sodium bicarbonate and water. After trituration with a mixture of ethyl acetate and hexane, 2.2 g of phenylmethyl ester of homogeneous [3- (benzoylamino) -2-oxo-4-phenylbutyl] methylcarbamic acid are obtained; melting point of 140 -141 C.
 EMI95.1
 b) () -N- [3-methylamino) -2-oxo-1- (phenylmethyl) propyl] hydrochloride [3-carbamic acid phenylmethyl ester (0.5 g) is dissolved in ethanol (50 ml) containing IN hydrochloric acid (2 ml). 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 so as to obtain 300 mg of () -N- [3- (methylamino) -2-oxo-1 hydrochloride - (phenylmethyl) propyl] benzamide in the form of a homogeneous white powder.
 EMI95.2
 c) Ester 1, 1- of (Z) amino) dimethylethylOne is stirred at room temperature under nitrogen overnight a reaction mixture of (¯) -N- [3- (methylamino) -2-oxo-1- hydrochloride

  <Desc / Clms Page number 96>

 (phenylmethyl) propyl] benzamide (1.65 gr; 5 mmol) of l- (bromoacetyl) -Lproline 1,1-dimethylethyl ester (2.9 gr; 10 mmol) of Example 68 (a), and diisopropylethylamine (1.74 ml; 10 mmol) in dimethylformamide (20 ml).

   The reaction mixture is partitioned between water and 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). Rapid chromatography (150 gr of Merck 60 silica gel) gives 0.7 gr of 1,1-dimethylethyl ester of (1) [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N -methylglycyl] -Lproline in the form of a yellow foam. d) (+) - 1- [N- [3- (benzoylamino)) hydrochloride -
 EMI96.1
 2-oxo-4-phenylbutyl] -N-methvlqlycyl] -L-proline.



  The ester from paragraph (c) (0.4 gr; 0.79 mmol) is mixed with 2N hydrochloric acid and 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 times) to give 0.28 g of () -1- [hydrochloride] N- [3- (benzoyiamino) -2-oxo-
 EMI96.2
 4-phenylbutyl] -N-methylglycyl] -L-proline in the form of a white solid; melting point of 125-130 C. Rf of 0.73 (silica gel; n-butanol / acetic acid / water; 3/1/1).
 EMI96.3
 Y Analysis: calculated for 25H29N305
C: 61.01; H: 6.24; N: 8.54 Found: C: 61.01; H: 6.10; N: 8.36.

  <Desc / Clms Page number 97>

 
 EMI97.1
 



  Example 92.



  (S) amino) 4dithia-7-azaspiro 4] -nonane-8-carboxylic acid trifluoroacetate salt.



   -7 - [[[3- (benzoyl- (1/1) a) Acid (S) -7- (bromoacetyl) -1, 4-dithia-7-azaspiro- [4. 4] nonane-8-carboxylic.



   Acid hydrochloride (S) -1, 4dithia-7-azaspiro is dissolved [4. 4] nonane-8-carboxylic (2, lgr; 8.7 mmol) in IN sodium hydroxide (25 ml), the mixture is cooled to OOC and bromide is added dropwise over 10 minutes bromoacetyl (2.1 gr; 0.91 ml; 10.4 mmol). The reaction mixture is stirred for 2 hours (OOC to room temperature), washed with ethyl acetate (2 times), the aqueous layer is acidified to pH 2 and extracted with l 'ethyl acetate (3 times).



  * The combined organic extracts are dried (Na2SO4) and concentrated to give 2.25 g of acid (S) -7- (bromo-
 EMI97.2
 acetyl) b) Diphenylmethyl ester of (S) -7- 1,4-dithia-7-azaspiro acid [4.



  Acid (S) -7- 1,4-dithia-7-azaspiro is dissolved [4.



  (2.25 gr; 6.9 mmol) in ethyl acetate (150 ml). 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 (2 times), 10% potassium bisulfate (2 times) and water, dried (Na SO) and

  <Desc / Clms Page number 98>

 it is concentrated so as to obtain 2.5 g of diphenylmethyl ester of (S) -7- (bromoacetyl) -1,4dithia-7-azaspiro acid [4. 4] nonane-8-carboxylic acid in the form of a white solid residue. c) Diphenylmethyl ester of (S) -7- [[[3- (benzoylamino) -2-oxo-4-phenylbutyl] methylamino] acetyla 1, 4-dithia-7-azaspiro [4. 4] nonane-8-carboxylic.



   A mixture of the paragraph ester product is stirred overnight at room temperature
 EMI98.1
 (b) (2.4 gr; 4.87 mmol), () - (0.81 gr; 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 times 100 ml).



  The organic extracts are washed. combined with saturated sodium bicarbonate, 10% potassium bisulfate and water, they are dried (Na2SO4) and concentrated to a dark oily residue (2.8 g). Rapid chromatography (200 g of Merck silica gel; 10% ethyl acetate / methylene chloride; 20% methanol / ethyl acetate) gives 1.1 g of ester
 EMI98.2
 (S) -7- 2-oxo-4-phenylbutyl] methylamino] 4-dithia- [[3- (benzoylamino) -7-azaspiro diphenylmethyl acid [4. 4] -nonane-8-carboxylic in the form of a yellow oil.
 EMI98.3
 d) Acid trifluoroacetate salt (S) -7-amino) 4-dithia- 7-azaspirol 4] -nonane-8-carboxylic

  <Desc / Clms Page number 99>

 [[[3- (benzoyl-The ester product from paragraph (c) is added (10.5 g;

   0.72 mmol) to cooled trifluoroacetic acid (2 ml) (OOC) containing anisole (0.1 ml).



  After stirring for 1 hour, the volatiles are separated in vacuo and the residue is treated with toluene (2 times). The oily residue is triturated with ether (4 times), which gives 0.36 g of acid trifluoroacetate salt (S) -7 - [[[3- (benzoylarnino) - 2-oxo-4- phenylbutyl) methylamino] acetyla, -1, 4-dithia-7azaspiro [4. 4] nonane-8-carboxylic (1/1); melting point of 120 -125 C. Rf of 0.45 (spreading) (silica gel; n-butanol / acetic acid / water; 4/1/1).



  Analysis: calculated for C27H31N3O5S2. C2HF3O2:
C: 53, 11; H: 4.92; N: 6.40; S: 9.78 Found: C: 52p8; H: 5.03; N: 6.53; S: 9.97
Example 93.



  Acid monohydrochloride (S) -2 - [[[3- (benzoylamino) -2- oxo-4-phenylbutyl) methylamino] acetyl] -1,2,3,4-tetrahydro-
 EMI99.1
 3-isoquinoéLne-carboxylic. a) 1,1-Dimethylethyl ester of 2- (bromoacetyl) 1,2,3,4-tetrahydro-3-isoquinoEinecarboxylic acid.



  To a cooled solution of 1,1-dimethylethyl ester of 1,2,3,4,4-tetrahydro-3-isoquinonecarboxylic acid (7.7 g; 33 mmol) in methylene chloride (100 ml) , diisopropylethylamine (6.23 ml; 36.3 mmol) is added and finally, over a period of 15 minutes, romoacetyl bromide (6.6 gr; 2.87 ml; 33 mmol) while maintaining the temperature at 5 vs.
 EMI99.2
 



  The reaction mixture is stirred overnight (-5 ° C. to room temperature), concentrated to approximately 33 1/3% of its volume, diluted with ethyl acetate (loo ml) , washed with saturated sodium bicarbonate (2 times), 10% potassium bisulfate (2 times) and water 2 times), dried

  <Desc / Clms Page number 100>

 (Na2SO4) and concentrated to a dark yellow semi-solid residue (11.0 g).

   Recrystallization from a mixture of ethyl acetate and hexane gives 4.2 g of 1,1-dimethylethyl ester of 2- (bromoacetyl) acid.
 EMI100.1
 1, 2, 3, 4-tetrahydro-3-isoquinonecarboxylic in the form of a cream-colored solid; melting point of 20-950C (850C). b) 1,1-Dimethylethyl ester of (S) (benzovlamino) -2-oxo-4-phenylbutyl] 1,2,3,4-tetrahVdro-3-isoguino-'Einecarboxylic ester.



   -2 - [[[3-Add (¯) -N- [3- (methylamino) -2-oxo-1- (phenylmethyl) propyl) benzamide (2.0 gr; 6 mmol) prepared as described in Example 91 (b), and diisopropylethylamine (0.77 g; 1.04 ml; 6 mmol)
 EMI100.2
 to a solution of 1,1-dimethylethyl ester of 2, 2, 3, 4-tetrahydro-3-isoquinoline carboxylic acid / 2, 12 gr; 6 mmol) in dimethylformamide (20 ml). the reaction mixture is stirred overnight, poured into water (50 ml) and
 EMI100.3
 the extract with ethyl acetate (3 times 50 ml).



  The ethyl acetate extracts combined with saturated sodium bicarbonate (2 times) and water (2 times), dried (Na2SO4) and concentrated in a yellow oily residue (2.9 g). Rapid chromatography (200 g of Merck silica; 2% methanol / chloroform) gives 0.68 g of 1,1-dimethylethyl ester
 EMI100.4
 (S) methylamino acid] 2, 3, 4-tetrahydro-3-isoquinoline-carboxylic acid in the form of a yellow desiccated foam.

  <Desc / Clms Page number 101>

 
 EMI101.1
 c) Acid hydrochloride (S) -2- 2-oxo-4-phenYlbutyl] methylamino] acetyl] -1, tetrahydro-3-isoquinoline carboxylic.



   The ester product of paragraph (b) (1.67 gr; 1.17 mmol) is treated with a mixture of 2N hydrochloric acid and acetic acid (5 ml). After stirring for 1 hour at room temperature, the reaction mixture is concentrated under reduced pressure and the resulting oily residue is triturated with ether (4 times) to give 0.55 g of acid monohydrochloride (S) - 2- [[[3- (benzoylamino) -2-oxo-4-phenylbutyl] methylamino] acetyl] -1, 2,3, 4-tetrahydro- 3-isoquinoline carboxylic as a slightly dirty white solid ; melting point 1230-126 C. Rf 0.47 (silica gel; n-butanol / acetic acid / water; 4/1).



  Analysis: calculated for C30H31N305. HCl. 0.32 HO:
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] -1- [N- [3- (benzoylamino) - 2-oxo-4-phenylbutyl] -N-methyglycyl] -4- (phenylthio) -Lproline hydrochloride. a) (4S) -1- (bromoacetyl) -4- (phenylthio) -L- proline.



   Bis (trimethylsilyl) acetamide (7.35 ml; 30 mmol) is added to a suspension of (4S) -4- (phenylthio) -L-proline (2.2 g; 10 mmol) in methylene chloride (juice ml; freshly distilled). The reaction mixture is stirred at room temperature for 2 hours until it becomes almost clear.



  The reaction mixture is then cooled to

  <Desc / Clms Page number 102>

   - 5 C and bromoacetyl chloride (1.9 g; 1.0 ml; 12 mmol) is added dropwise while maintaining the temperature at -5 C. After stirring overnight (from -5 C to at room temperature), the reaction mixture is concentrated to about 50% of its volume, partitioned between saturated sodium bicarbonate and ethyl acetate and the layers are separated. The organic layer is extracted once again with saturated sodium bicarbonate. The combined aqueous layers are acidified to pH 2.0 with 10% potassium bisulfate and extracted with ethyl acetate (3 times).

   Fractions
 EMI102.1
 with ethyl acetate are combined, dried and concentrated to give 3.5 g of (4S) -1- (bromoacetyl) -4- in the form of a viscous oil. b) (4S) -1- (bromoacetyl) - 4- (phenylthio) -L-proline diphenylmethyl ester.



   A solution of diphenyldiazomethane (2.0 g; 10.2 mmol) in water is added dropwise.
 EMI102.2
 ethyl acetate (50 ml) to a solution of (. S) (bromoacetate) -4- (3.5 g; -1-10.2 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 (2 times) and water (2 times), dried (Na2SO4) and concentrated to give 4.78 g of (4S diphenylmethyl ester ) -1- (bromacétyl) -4- (phnéylthio) -L-proline in the form of a yellow viscous oil.

  <Desc / Clms Page number 103>

 c) [1 (-), 4S] -1- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N-methylglycycl] -4- (phenylthio) -L-proline diphenylmetylcrue ester.



   () -N- [3- (methylamino) -2-oxo- 1- (phenylmethyl) propyl] benzamide (2.42 g; 7.2 mmol) prepared as described in Example 91 (b) is added and diisopropylethylamine (0.93 g; 1.25 ml; 7.2 mmol) to a solution of (4S) -1- (bromoacetyl) -4- (phenylthio) -L-proline diphenylmethyl ester (4, 78 gr; 9.4 mmol) in dimethylformamide (20 ml).



  After stirring overnight at room temperature, the reaction mixture is poured into water (50 ml) and extracted with ethyl acetate (3 times). The combined ethyl acetate extracts are washed with saturated sodium bicarbonate (2 times) and water (2 times), dried (Na2SO4) and concentrated in a yellow oil (6.2 gr).

   Rapid chromatography (Merck silica gel; 2% methanol / methylene chloride) gives 3.2 g of [l (¯), 4S] -l- [N- [3- (benzoylamino) - 2-diphenylmethyl ester. -oxo-4-phenylbutyl] -N-methylglycyl] -4- (phenylthio) -Lproline in the form of a pale yellow foam. d) M (), 4S] -1 hydrochloride -1- [N- [3- (benzoyl-amino) -2-oxo-4-phenylbutyl] -N-methylglycycl] -4- (phenylthio) -L-proline.



   The ester product of paragraph (c) (1.6 g; 2.2 mmol) is treated with a mixture of 2N hydrochloric acid and 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

  <Desc / Clms Page number 104>

 ether overnight to give 1.2 g of [1 (), 4S] -1- [N- [3- (benzoylamino) -2- oxo-4-phenylbutyl] -N-methylglycyl] -4 monohydrochloride - (phenylthio) -Lproline in the form of a slightly dirty white solid; melting point of 131 -133 C; 0.38 rf
 EMI104.1
 (silica gel; n-butanol / acetic acid / water; 4/1/1).



  1HN0S.HCl.0,9HO C: 60, 82; H: 5.90; N: 6.87; S: 5.24; Cl: 5, 79 Found:: 60, 82 H: 5, 74; N: 6.84; S: 5.25; Cl: 5, 75.



  Example 95.



  (4S) -l- oxo-4-phenylbutyl] -N-methylglycyl] monohydrochloride -4- Analysis: calculus for GL-proline. a) 1,1-Dimethylethyl ester of (4S) -1- (bromoacetyl) - 4- (fluorophenoxy) -L-proline.



   Bromoacetic acid (1.4 g; 7.5 mmol), hydroxybenzotriazole hydrate (1.14 gr; 7.5 mmol) and dicyclohexylcarbodiimide (1.54 g; 7.5 mmol) are added. ) to a solution of 1,1-dimethylethyl ester of (S) -4- (fluorophenoxy) -L-proline (2.1 gr; 7.5 mmol) in distilled tetrahydrofuran (50 ml). The reaction mixture is stirred overnight, the precipitated dicyclohexylurea is filtered off and the filtrate is concentrated.

   The residue is dissolved in ethyl acetate (50 ml) and washed with saturated sodium bicarbonate (2 times), 10% potassium bisulfate (2 times) and water (2 times ), it is dried (Na2SO4) and concentrated to give 3.1 g of 1,1-dimethylethyl ester of (4S) -1- (bromoacetyl0-4- (fluorophenoxy) -Lproline in the form of a residue oily.

  <Desc / Clms Page number 105>

 
 EMI105.1
 d) Ester of (4S) -1- (benzoylamino) 4- (4-fluorophenoxy) Is added () l- (2.83 gr; 8.5 mmol) prepared as described in Example 91 (b) and diisopropylethylamine (1.43 gr; 1.92 ml; 11.0 mmol) to a solution of 1,1-dimethylethyl ester of (4S) -
1- (bromoacetyl) -4- (fluorophenoxy) -L-proline (3.4 gr;
8.5 mmol) in dimethylformamide (20 ml).

   After stirring overnight at room temperature, the reaction mixture is poured into water (100 ml) and extracted with ethyl acetate (3 times).



   The combined ethyl acetate extracts are washed with saturated sodium bicarbonate (2 times), 10% potassium bisulfate (2 times) and water (2 times), dried (Na2SO4) and concentrated into a dark colored residue (5.5 gr). Rapid chromatography (silica gel LPS-1; 50% ethyl acetate / methylene chloride) gives 1.2 g of 1,1-dimethylethyl ester of (4S) -l- [N- [3- (benzoylamino) -2- oxo-4-phenylbutyl] -N-methylglycyl] -4- (4-fluorophenoxy) -
L-proline in the form of a pale yellow foam. c) (4S) -1- [3- (benzoylamino) -2-oxo- hydrochloride
4-phenylbutyl] -N-methylglycyl] -4- (4-fluorophenoxy) -L-proline.



   The ester product of paragraph (d) (1.2 g; 1.95 mmol) is treated with a mixture of 2N hydrochloric acid and 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

  <Desc / Clms Page number 106>

 
 EMI106.1
 overnight to give 0.92 g of (4S) N-methylglycyl] -4- monohydrochloride as a slightly dirty white solid; melting point 131-140 C. Rf 0.54 (silica gel; n-butanol / acetic acid / water; 3/1/1).
 EMI106.2
 



  Analysis: for C j-L Jz J b
C: 61.24; H: 5.65; N: 6.91; Cl: 5.83 Found: C: 61.24; H: 5.66; N: 7.09; Cl: 5.70.



   Similarly, the process 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- monohydrochloride
 EMI106.3
 phenylbutyl] -N-methvlqlycvl] -N-cyclohexylqlycine.



  1,1-Dimethylethyl ester of () -N- -N- [3-amino) -2-oxo-4-phenylbutyl] -N-methylqlycine.



  Bromoacetic acid 1,1-dimethylethyl ester (13.8 g; 0.15 mi; 19.5 mmol) and diisopropylethylamine (2.5 g; 3.4 ml; 19.5 mmol) are added. ) to a solution of () -N- [3- 2-oxo-1- (5.0 g; 15 mmol) prepared as described in Example 91 (b) in dimethylformamide (20 ml). After stirring overnight at room temperature, the reaction mixture is poured into water (100 ml) and extracted with ethyl acetate (3 times).

   The combined ethyl acetate extracts are washed with saturated sodium bicarbonate (2 times), 10% potassium bisulfate (2 times) and water (2 times), dried (Na SO ) and concentrated in a yellow oil, which becomes

  <Desc / Clms Page number 107>

 a foam dried by drying in a high vacuum to give 5.4 g of 1,1-dimethylethyl ester of (¯) -N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N-methylglycine. b) (-) - N- [3- = (benzoyiamino) -2-oxo-4-phenylbutyl] -N-methylqlycine hydrochloride.



   The ester product of paragraph (a) (4.51 gr; 11 mmol) is treated with a mixture of 2N hydrochloric acid and acetic acid (20 ml). After stirring for 2.5 hours at room temperature, the reaction mixture is concerted under reduced pressure and the oily residue is triturated with ether
 EMI107.1
 to give 3.3 g of (-) - N- (benzoylamino) monohydrochloride in the form of a slightly dirty white solid. c) 1,1-Dimethylethyl ester of N- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N-methylglycyl] -N-cyclohexylqlycine.



   Is added 1,1-dimethylethyl ester of N-cyclohexylglycine (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) at one
 EMI107.2
 solution of () -N- -N- [3-oxo-4-phenylbutyl] -N-methylglycine hydrochloride (1.0 g; 2.6 mmol) in distilled tetrahydrofuran (50 ml). The reaction mixture is stirred overnight, the precipitated dicyclohexylurea is filtered off and the filtrate is concentrated.

   The residue is dissolved in ethyl acetate (50 ml) and washed with saturated sodium bicarbonate (2 times), potassium bisulfate at

  <Desc / Clms Page number 108>

 10% (2 times) and water (2 times), dried (Na2SO4) and concentrated in an oily residue (1.5 gr). Rapid chromatography (100 gr; Merck 60 silica gel) gives 0.49 gr of 1,1-dimethylethyl ester of N- [N- [3- (benzoylamino) -2, oxo-4-
 EMI108.1
 phenylbutyl] -N-methylglycyl] -N-cyclohexylglycine in the form of a foam. d) N- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N-methylqlycyl] -N-cyclohexylqlycine hydrochloride.



   The ester product of paragraph (c) (0.48 g; 0.87 mmol) is treated with a mixture of 2N hydrochloric acid and acetic acid (10 ml). After stirring for 2 hours at the amliant temperature, the reaction mixture is concentrated under reduced pressure and the oily residue is triturated with ether overnight to give 0.32 g of mono-
 EMI108.2
 N-butyl] -N-methylglycyl] -N-cyclohexylglycine hydrochloride as a slightly dirty white solid; melting point of 1310-14 SoC. Rf 0.36 (silica gel; n-butanol / acetic acid / water; 4/1/1).
 EMI108.3
 



  Analysis: calculation for C-HN0.HC1.0,7HO
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.



  (S) acid methyl ester -7 - [[[(¯) -3- (benzoylamino) -
 EMI108.4
 2-oxo-4-phenylbutyl] methylamino] azaspiro 4] -nonane-8-carboxyli acetyl] -l-dithia-7-Add methyl acid ester monohydrochloride (S) -1,4-dithia-7 -azaspiro [4. 4] - nonane-8-carboxylic (0.66 gr; 2.5 mmol), dicyclohexylcarbodiimide (0.54 gr; 2.5 mmol),

  <Desc / Clms Page number 109>

 hydroxybenzotriazole hydrate (0.39 g; 2.5 mmol) and diisopropylethylamine (0.9 ml; 5 mmol) to a solution of (I) -N- [3- (benzolyamino) monohydrochloride -2-oxo-4-phenylbutyl] -N-methylglycine (1.0 g; 2.5 mmol) prepared as in Example 96 (b), in distilled tetrahydrofuran (50 ml).

   The reaction mixture is stirred overnight, the precipitated dicyclohexylurea is filtered off and the filtrate is concentrated. The residue is dissolved in ethyl acetate (100 ml) and washed with saturated sodium bicarbonate (2 times) and water (2 times), dried (Na2 so4) and dried. concentrated in a yellow oily residue (1.3 gr).

   Rapid chromatography (Merck silica gel; 25% ethyl acetate / methylene chloride; 1% methanol / methylene chloride) gives 0.53 g of acid methyl ester
 EMI109.1
 (S) methylamino] acetyl-1, 4-dithia-7-azaspiro [4. carboxylic in the form of a white foam; melting point of 60 -62 C. Rf of 0.52 (silica gel; 5% methanol / methylene chloride).
 EMI109.2
 N305B.0, 2o Jj J 5 2
C: 59.87; H: 6.04; N: 7.48; S: 11.42 Found: C: 59.87; H: 5.94; N: 7.56; S: 11.36.



   Example 98.



  (S) -7- acid methyl ester monohydrochloride
 EMI109.3
 [) -3- methylamino] acetyl] -1, [[(carboxylic.



   The methyl ester product of Example 97 (0.26 g; 0.46 mmol) is treated with a mixture

  <Desc / Clms Page number 110>

 2N hydrochloric acid and acetic acid until it becomes homogeneous (2 minutes), it is concentrated under reduced pressure and the oily residue is triturated with ether (2 times) to obtain 0.26 gr of (S) -7- [[[(-) - 3- (benzoylamino) -2-oxo-4-phenylbutyl] methylamino] methyl ester monohydrochloride hydrochloride] - acetyl] -1, 4-dithia-7 -azaspiro [4. 4] nonane-8-carboxyli-
 EMI110.1
 as in the form of a white solid; melting point of 790-850C. Rf 0.53 (methanol silica gel / methylene chloride).



  Analysis: calculated for 0S. zo JJ J 5 z
C: 55.84; H: 5.88; N: 6.98; S: 10.64; Cl: 5.88 Found: C: 55.84; H: 5.95; N: 6.78; S: 10.43; Cl:. 5.66.



   Example 99, Methyl ester hydrochloride, (4S) -1- [N- [3-
 EMI110.2
 (benzoylamino) -2-oxo-4-phenylbutyl] -N-methylqlycyl] - 4- a) (4) 74- (fluorophenoxv) -L-proline methyl ester monohydrochloride.



   Thionyl chloride (8.09 ml; 11 mmol) is added to a suspension of (4S) -4- (fluorophenoxy) - L-proline (2.5 g; 11 mmol) in methanol at -30 ° C under a argon atmosphere. The reaction mixture is stirred at -20 ° C for 2 hours, and then at room temperature for 16 hours. The solvent is separated at reduced pressure, the residue is redissolved in methylene chloride (150 ml) and washed with IN sodium carbonate (2 times) and water (2 times). After drying (MgSO), an excess of hydrochloric acid and methanol is added and the solvent is separated at reduced pressure. Addition of ether

  <Desc / Clms Page number 111>

 gives a light brown solid (2.6 gr).

   Recrystallization from a mixture of methanol and ether gives 1.49 g of (4S) -4- (fluorophenoxy) -L-proline methyl ester monohydrochloride in the form of a solid
 EMI111.1
 light brown; melting point of 147 -148 C; [a] D '+ D (c = 1.55; methanol). b) (4S) -1- [N- [3- (benzoylamino) -2oxo-4-phenylbutyl] -N-methylglycycl] -4- (4-fluorophenoxy) L-proline methyl ester.



   (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;
 EMI111.2
 3 mmol) to a solution of (i) N- [3-glycine monohydrochloride (1.17 g; 3 mmol) prepared as described in Example 96 (b) in distilled tetrahydrofuran (20 ml). The reaction mixture is stirred overnight, the precipitated dicyclohexylurea is filtered off and the filtrate is concentrated. The residue is dissolved in ethyl acetate (50 ml), washed with saturated sodium bicarbonate (2 times) and water (2 times), dried (Na2SO4) and concentrated into an oily residue (1.1 gr}.

   Rapid chromatography (200 gr; Merck 60 silica gel; 3% metha-
 EMI111.3
 nol / chloroform) gives 0.15 g of methyl ester of (4S) -1- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N-methylglycyl] -4- (4-fluorophenoxy) -L -proline in the form of a foam. c) (4S) -1- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -N-methylglycyl] methyl hydrochloride ester hydrochloride -

  <Desc / Clms Page number 112>

 4- (4-fluorophenoxy) 2-proline.



   The methyl ester product of paragraph (b) (0.15 g; 0.26 mmol) is treated with a mixture of 2N hydrochloric acid and acetic acid until homogeneous (2 minutes), it is concentrated under reduced pressure, and the oily residue is triturated with ether (2 times) to obtain 0.14 g of monochlor-
 EMI112.1
 (4S) -1-2-oxo-4-phenylbutyl] -N-methylglycyl] -4-phenoxy) -L-proline methyl ester hydrate as a slightly dirty white solid; melting point of 1050-1250C.



   Rf 0.27 (silica gel; 5% methanol / chloroform).



  Analysis: calculated for C32H34F06. HCl
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) -1- [N [(S) -3- (benzovlamino) -2- oxo-4-phenylbutyl] -L-alanyl] -4- (4-fluorophenoxy) -L-proline hydrochloride. a) Ester 1,1-dimethylethylicrue of (S) -N- [3- (benzovlamino-2-oxo-4-phenylbutyl] -L-alanine.



   1.1-dimethylethyl ester hydrochloride of L-alanine (6.0 gr; 33.1 mmol), sodium bicarbonate (6.1 gr; 72 mmol) and sodium iodide (4.9 gr; 33.1 mmol) to a stirred solution of (S) -N- [3-chloro-2-oxo-1- (phenylmethyl) propyl] benzamide (10.0 gr; 33.1 mmol) in dimethylformamide (80 ml). The resulting solution is stirred overnight at room temperature, poured into ether and washed with water

  <Desc / Clms Page number 113>

 (2 times) and 10% sodium bicarbonate. The ethereal solution is extracted with IN hydrochloric acid (3 times), the combined extracts are made basic by the addition of solid sodium bicarbonate and they are extracted with ethyl acetate (4 fds).



  The organic extracts are combined, dried (MgSO.) And concentrated to give 8; 9 gr of a pale yellow solid. A portion of this material is recrystallized from ethyl acetate to give 1,1-dimethylethyl ester of (S) -N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -L -alanine in the form of a white solid; melting point 106.5 -110 C. b) (S) -N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -L-alanine hydrochloride.



   A solution of the ester product of paragraph (a) (2.95 g; 5.4 mmol) in 1, 4N hydrochloric acid in acetic acid (39 ml) is stirred at ambient temperature for 2 hours. ). The resulting white precipitate is collected, rinsed with ether and dried to give 2.27 g of (S) -N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -L monohydrochloride - alanine; melting point of 208-209 C (decomposition); [a] = -71 (c = 0.38% in methanol). Rf 0.51 (sil2 ge2; chloroform / methanol / acetic acid; 4/1/1).



  Analysis: calculated for C20H22N204. HCl:
C: 61.64; H: 5.93; N: 7.17; Cl: 9.07 Found: C: 61.33; H: 5.97; N: 7.17; Cl: 8.79.
 EMI113.1
 c) (S) -N- [Triethylamine (2.1 ml;

  <Desc / Clms Page number 114>

 [N- (benzoylamino) -2-oxo-4-phenylbutvl] -N-15 mmol) to a mixture of (S) -N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -L monochlorhyrate -alanine (2.0 gr; 5.1 mmol), benzyl chloroformate (730 gl; 5.1 mmol), water (7 ml) and dioxane (7 ml) at 25 C. The mixture is stirred resulting at 25 ° C for 3 hours, after which it is poured into a 5% aqueous solution of sodium bicarbonate and washed with ether. The aqueous layer is acidified (HCl) and extracted with ethyl acetate (3 times).

   The extract is dried (MgSO.) And concentrated to obtain a colorless oil. Trituration with ether produces a white granular solid (150 mg) which is collected and discarded. The liquor is concentrated under vacuum to obtain 1.75 g of (S) -N- [N-
 EMI114.1
 (benzoylamino) carbonyl] -L-alanine in the form of a glassy white solid. d) Phenylmethyl ester of (4S) -l- [N- [(S) -3- (benzoyl-
 EMI114.2
 amino) L-alanyl] -4- <4-fluorophenoxy) -L-proline.



  A mixture of (S) nylmethoxy) carbonylj-L-alanine (300 mg; 0.62 mmol), of p-toluenesulfonic acid salt of (4S) -4- (4S) -4- phenyl ester is stirred at 250 ° C. for 20 hours. 4-fluorophenoxy) -L-proline (300 mg; 0.62 mmol), triethylamine (90 µl; 0.62 mmol), dicyclohexylcarbodiimide (130 mg; 0.62 mmol) and hydroxybenzotriazole hydrate (90 mg; 0.62 mmol). in tetrahydrofuran (7 ml). The mixture is then filtered and diluted with ethyl acetate.

   The resulting solution is washed sequentially with

  <Desc / Clms Page number 115>

 IN hydrochloric acid and a 10% aqueous solution of sodium bicarbonate, dried (MgSO.), filtered and concentrated to obtain 500 mg of ester
 EMI115.1
 (4S) 2-oxo-4-phenylbutyl] -N- [alanyl] -4- phenylmethyl form of a pale yellow oil. e) (4S) -1- [N - [(S) -3- (benzoylamino) - 2-oxo-4-phenylbutyl] -L-alanyl] -4- (4-fluorophenoxy) -Lproline hydrochloride.



   A mixture of the ester product of paragraph (d) (500 mg; 0.6 mmol), palladium on carbon catalyst (10%; 100 mg), absolute ethanol (15 ml) and aqueous hydrochloric acid 1, ON (8001; 0.8 mmol) is hydrogenated at one atmosphere and at 250C for 17 hours, after which it is filtered and concentrated. The residue is chromatographed on HP-20 using a linear gradient from [aqueous hydrochloric acid 0, OlN / methanol 9/1] to [aqueous hydrochloric acid O, OlN / methanol 1/1]. The fractions containing the desired product (thin layer chromatography) are combined and concentrated. The residue is dissolved in a minimum amount of methanol.

   Ether is added, which gives a white precipitate which is collected and dried under vacuum to obtain 200 mg of (4S) -1- [N- [(S) -3- ( benzoyl-
 EMI115.2
 amino) L-proline; melting point of 152 -153 C (decomposition); 1a] (c = 0.5, methanol). 0.75 (silica gel; chloroform / methanol / acetic acid; 4/1/1).

  <Desc / Clms Page number 116>

 
 EMI116.1
 Analysis: calculationforCHFN-0.



  J-L 32 J 6 2
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.



  [L (S), 4R] -l -'- N- [3- (benzoylamino) - monohydrochloride -
 EMI116.2
 2-oxo-4-phenylbutyl] -L-alanyl] -4-phenyl-L-proline. a) Succinimido ester of (S) -N- oxo-4-phenylbutyl] -N- [[N- (benzoylamino) -2-alanine.



   Stirred at 1250C for 18 hours a mixture of (S) -N- [N- (benzoylamino) -2-oxo-4-phenylbutyl] N - [(phenylmethoxy) carbonyl] -L-alanine (800 mg; 1, 6 mmol) prepared as described in Example 100 (c), of dicyclohexylcarbodiimide (340 mg; 1.6 mmol) and N-hydroxysuccinimide (190 mg; 1.6 mmol) in tetrahydrofuran (5 ml). After that, it is filtered and concentrated to obtain 950 mg of ester
 EMI116.3
 (S) -N- phenylbutyl] -N- succinimido b) [l- (S), 4R] -1- [N- butvl] -N- [N- (benzoylamino) -2-oxo-4-L- proline.



   (4R) -4-phenylL-proline hydrochloride (375 mg; 1.7 mmol) and triethylamine (40 μl; 3.2 mmol) are added to a solution of succinimido ester of (S) -N- [N- (benzoylamino) -2-oxo-4-phenylbutyl] -N- f (phenylmethoxy) carbonyl] -L-alanine (950 mg; 1.6 mmol) in dimethylformamide (5 ml). The resulting mixture was stirred at 25 ° C for 24 hours, after which it was poured into excess IN hydrochloric acid and extracted with ethyl acetate (3 times). The

  <Desc / Clms Page number 117>

 
 EMI117.1
 extracts are combined, dried), filtered and concentrated to obtain 1.1 g of [1 (S), 4R] -1- [N- [3- (benzoylamino) carbonyl] -L-alanyl] -4-phenyl-proline . c) [1 (S), 4R] -1- [N- [3-amino] -2-oxo-4-phenylbutyl] -L-alanvl] -4-phenyl-L-proline hydrochloride.



   The product of paragraph (b) (1.0 gr; 1.5 mmol), ethanol (20 ml), water (5 ml), hydrochloric acid 1, ON (1.5 ml ; 1.5 mmol) and palladium on carbon catalyst (10%; 100 mg) are hydrogenated at one atmosphere and at 250C for 18 hours, after which it is filtered and concentrated. The residue is chromatographed on HP-20 using a linear gradient [aqueous hydrochloric acid 0, OlN / methanol; 40/60 to 10/90]. The fractions containing the desired product are combined (thin layer chromatography) and concentrated. The residue is dissolved in a minimum amount of methanol.

   Ether was added, the resulting white precipitate was collected and dried to give 300 mg of [1 (S), 4R] -1- [N- [} -enzoylamino) -2-oxo- monohydrochloride
 EMI117.2
 4-phenylbutyl] -L-alanylJ-4-phenyl-L-proline, melting point 160 -162 C (decomposition); [a] = -57 D (c = 1.5; methanol). Rf 0.8 (silica gel / chloroform / methanol / acetic acid; 4/1/1.
 EMI117.3
 Analysis: calculated
C: 63.27; H: 6.29; N: 7.14; Cl: 6.02 Found C: 63.27; H: 6.17; N: 7.19; Cl: 5.97.

  <Desc / Clms Page number 118>

 



   Example 102.



  [1 (S), 4S] -1 hydrochloride -1- [N- [3- (benzoylamino) -
 EMI118.1
 2-oxo-4-phenylbutyl] -L-alanyl] -4-phenyl-L-proline.



  Following the method of Example 101 but using (4S) -4-phenyl-L-proline hydrochloride in paragraph (b), [1 L-alanyl-4-phenyl-L- monohydrochloride is obtained proline; melting point of 1380-143 C; [a] D = -61 (c = 0.3% in methanol).



     Rf 0.84 (silica gel; chloroform / methanol / acetic acid; 4/1/1).
 EMI118.2
 



  Analysis: calculationforC-.HN0.HC1.2,13HO
C: 61.80; H: 6.35; N: 6.98; Cl: 5.88 Found: C: 61.80; H: 6.06; N: 7.05; Cl: 5.65
Example 103.



  [1 (S), 4R] -1 hydrochloride -1- [N- [3- (benzoylamino) -
 EMI118.3
 2-oxo-4-phenylbutyl] -L-alanyl] -4-cyclohexyl-L-proline.



  Following the method of Example 101 but using (4R) -4-cyclohexyl-Lproline hydrochloride in paragraph (b), [1 (S), 4R] -l- phenylbutyl] monohydrochloride is obtained - L-alanyl] -4-cyclohexyl-L-proline; melting point of 140 -154 C (decomposition); lc -830 (c = 0.36% in methanol). Rf 0.84 (silica gel; chloroform / methanol / acetic acid; 4/1/1).



  Analysis: calculated for C31H39N305. HCl. 0.79 HO:
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.
 EMI118.4
 



  Example 104.



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

  <Desc / Clms Page number 119>

 



   Following the method of Example lol but using (4S) -4-cyclohexyl-L-proline hydrochloride in paragraph (b), mono-
 EMI119.1
 [1 (S) hydrochloride, 4SJ-1- [N- [3-oxo-4-phenylbutylJ-L-alanylJ-4-cyclohexyl-L-proline melting point 1390-1410C (decomposition); [a] D = -800 (c = 0.2% in methanol). Rf 0.83 (silica gel; chloroform / methanol / acetic acid; 4/1/1).
 EMI119.2
 



  Analysis: for CHN0.HC1.1, calculated OC: 62.28; H: 7.26; N: 7.03; Cl: 5.93 Found: C: 62.28; H: 7.01; N: 7.02; Cl: 6.16.



   Example 105.



  Acid monohydrochloride (S) -7- [(S) -2- [[3- (benzoylamino) -
 EMI119.3
 2-oxo-4-phenylbutvl] -amino] -1-oxopropyl] -1,4-dithia-7-azaspiro 4] nonane-8-carboxylic.



  Following the method of Example 101 but using acid hydrochloride (S) -1,4-dithia-7azaspiro [4. 4] nonane-8-carboxylic in the paragraph (b) for the reagent of L-proline, one obtains mono-
 EMI119.4
 acid hydrochloride (s) -7- 2-oxo-4-phenylbutyl] 4-dithia-7azaspiro [4. 170-172 C; (c = 1.4% in methanol).



  0.78 silica; chloroform / methanol / acetic acid; 6/1/1).



  2 / J-L 352
C: 54.77; H: 5.71; N: 7.10; 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 monochlorhyrate [l (S), 5S] -1- [N- [3- (benzoyl-

  <Desc / Clms Page number 120>

 
 EMI120.1
 amino) phenyl-1H-pyrazole-5-carboxylic.



   Following the method of Example 101 but using (S) -4, 5-dihydro-3-phenyl-1H-pyrazole-5-carboxylic acid for the L-proline reagent in paragraph (b) , the above-mentioned compound is obtained.



   Example 107.
 EMI120.2
 



  (S) -2- (Benzoylamino) 2, 3, 4-tetrahydro- [N- [(S) -3-3-isoquinoline carboxylic acid) monohydrochloride.



   Following the method of Example 101 but using acid hydrochloride (S) -1, 2,3, 4tetrahydro-3-isoquinoline carboxylic in paragraph (b) in place of the proline reagent, monohydrochloride is obtained (S) -2- [N- [(S) -3- (benzoyl- amino) -2-oxo-4-phenylbutyl] -L-alanyl] -1,2,3,4-tetra-
 EMI120.3
 hydro-3-isoquinoline carboxylic.



  Example 108.



  1- [N- [3- oxo-4-phenylbutyl] -L-alanyl] -2-2-hydroxyphenyl) -4 (benzoylamino) -2-thiazolidinecarboxylic acid monohydrochloride.



   Following the method of Example 101 but using 2- [(2-phenylmethoxy) phenyl] -4 (R) -thiazolidinecarboxylic acid hydrochloride in paragraph (b) in place of the proline reagent, after separation of the hydroxy protection group from the mono-
 EMI120.4
 1- [N- [phenylbutyl] -L-alanyl] -2- (S-3- (benzoylamino) -2-oxo-4-zolidinecarboxylic acid hydrochloride.



   Similarly, one can use the

  <Desc / Clms Page number 121>

 process of Examples 96, 97, 99 and 100 to 108 to prepare the compounds of Examples 1 to 95.



   Example 109.



  (I) -1- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -L-alanyl] -L-proline methyl ester monohydrochloride. a) 1,1-Dimethylethyl ester of 1- [N- (2-ethoxY-2-oxoethyl) -N- [(phenylmethoxy) carbonyl] -L-proline.



   We resume in tetrahydrofuran
 EMI121.1
 (400 ml) with stirring of the 1,1-dimethyl-ethyl ester of L-alanyl-L-proline (25, 44 g; 105 To this is added bromoethyl acetate (11.64 ml; 105 mmol) and diisopropylethylamine (18.3 ml; 105 mmol). After 6 hours, the reaction mixture is cooled in an ice bath and benzyl chloroformate (16.5 ml; 115.5 mmol) is added. and diisopropylethylamine (20.1 ml; 115.5 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, it is taken up in ethyl acetate and washed until neutral with 10% potassium bisulfate and saturated sodium bicarbonate. The crude product is purified (52.3 g) on a column of silica gel, eluting with a mixture of ethyl acetate and hexane (1/1) to give 42.4 g of 1,1-dimethylethyl ester of 1- [N- (2-ethoxy-2-oxoethyl) -N- [(phenylmethoxy) carbonyl] -L-alanyl] -L-proline. b) 1,1-dimethylethyl ester of 1- [N- (carboxymethyl) - N- [(phenylmethoxy) casrbonyl] -L-alanyl] -L-proline.

  <Desc / Clms Page number 122>

 



   The ester product from paragraph (a) is taken up (21 gr; 45.4 mmol) in methanol (150 ml) and IN sodium hydroxide (50 ml; 50 mmol) with stirring at room temperature. After 5.5 hours, the methanol is separated in vacuo and the aqueous portion is extracted with ethyl acetate.



  The aqueous portion is acidified with concentrated hydrochloric acid and extracted into acetate
 EMI122.1
 ethyl to give 16.95 g of l-carbonyl] -L-alanyl] -L-proline l, 1-dimethyl-ethyl ester. c) Ester 1, of (i) amino) [N- (carboxymethyl) -N- [(phenylmethoxy) -L-alanyl] -L-proline.



   The ester product from paragraph (b) (10.0 g; 23 mmol) is taken up in dry tetrahydrofuran (75 ml) with stirring in an ice bath. To this, 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 kept at room temperature for 1 hour. The mixture is then concentrated to dryness under vacuum and taken up in tetrahydrofuran (40 ml) with stirring in an ice bath. To this, 2-phenyl-4- (phenylmethyl) -5 (4H) -oxazolone (6 g; 23.8 mmol) is added dropwise in tetrahydrofuran (35 ml) followed by triethylamine (3.22 ml ; 23 mmoles). An additional amount of triethylamine is added to maintain a basic atmosphere.

   After 20 minutes, the ice bath is removed and the reaction mixture is maintained

  <Desc / Clms Page number 123>

 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 flow of argon. The reaction mixture is concentrated to dryness, it is taken up in ethyl acetate, and it is washed until neutral with saturated sodium bicarbonate and dilute hydrochloric acid.

   The crude product (12.8 g) is chromatographed on silica gel, eluting with a mixture of ethyl acetate and hexane (1/1) to obtain 9.05 g of ester l, l- dimethylethyl of
 EMI123.1
 (Z) [-1- [N- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -Nd) (¯) -1- [N- [N- [3- (benzoylamino) - 2-oxo-4-phenylbutyl] N- [(phenylmethoxy) carbonyl] -L-alanyl] -L-proline.



   The ester product from paragraph (c) (11.4 g; 17.7 mmol) is taken up in trifluoroacetic acid (50 ml) and kept at room temperature for 45 minutes. It is then concentrated to dryness and it is triturated with a mixture of ether and hexane to obtain 10.3 g of crude product. This material is purified on a column of silica gel in a mixture of benzene and acetic acid
 EMI123.2
 (8/2) to obtain 9.7 g of () -l- [N- amino) L-alanyl] -L-proline; melting point of 70 -91 C;

  <Desc / Clms Page number 124>

 
 EMI124.1
 23 D p -53, = 1.22; methanol). 0.41 (gelD f [N- [3- (benzoyl-silica; benzene / acetic acid / 8/2).



  Analysis: calculated for C33H35N307
C: 67.68; H: 6.02; N: 7.17 Found: C: 67.95; H: 6.02; N: 6.82 e) (¯) -1- [N- [N- [N- [3- (benzoylamino) - 2-oxo-4-phenylbutyl] -N- [(phenylmethoxy) carbonyl] -Lalanyl] methyl ester L-proline.



   The acid product from paragraph (d) (3.5 g; 6 mmol) is taken up in dimethylformamide (12 ml) with stirring at room temperature. To this is added sodium bicarbonate (630 mg; 7.5 mmol) and methyl iodide (0.47 ml; 7.5 mmol). After 18 hours, an additional amount of methyl iodide (7.4 mmloes) and sodium bicarbonate (7.4 mmol) is added. After stirring for another four hours, the reaction mixture is concentrated to dryness under vacuum, it is taken up in ethyl acetate and washed with saturated sodium bicarbond.

   The crude product (3.7 g) is purified on a column of silica gel in a mixture of ethyl acetate and hexane (2/1) to
 EMI124.2
 obtain 3.5 g of methyl ester of () (benzoylamino) -2-oxo-4-phenylbutyl] -N- -1- [N- [N- [3-carbonyl] -L-alanyl] -L-proline . f) Methyl ester hydrochloride of (i) -1- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -L-alanyl] -L-proline.



   The ester product from paragraph (e) is taken up (910 g; 1.5 mmol) in ethanol (95%; 100 ml) and in IN hydrochloric acid (1.8 ml).



  Palladium on carbon catalyst is added

  <Desc / Clms Page number 125>

   (10%; 180 mg) and the reaction mixture is stirred under hydrogen overnight. The reaction mixture is filtered to separate the catalyst, it is concentrated to dryness and lyophilized twice in water to obtain 700 mg of methyl ester monohydrochloride.
 EMI125.1
 (i) alanyl] -L-proline; melting point of 110 -128 C (900C); [a] = -69, (c = 1.18; methanol). Rf of D 0.63 (gel of; chloroform / methanol / acetic acid; 9/1/1).



  Analysis: calculationfor C.-H..N0. b DD-3 5 2
C: 60.22; H: 6.58; N: 8, 11; Cl: 6.84 Found: C: 60.22; H: 6.28; N: 8, 10; Cl; 7.06.



   Example 110 Monomethanesulfonate of () -l- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -L-alanyl] L-proline methyl ester.



   The methyl ester of () -1- [N- [N- [N- [3- (benzoylamino) 2oxo-4-phenylbutyl) is taken up in methanol (60 ml) and methanesulfonic acid (0.066 ml; 1 mmol). ] -N- [(phenylmethoxy) carbonyl] -L- alanyl] -L-proline (600 mg; 1 mmol), prepared as described in Example 109 (e). Palladium on carbon catalyst (10%; 120 mg) is added and the reaction mixture is stirred under hydrogen for 2.5 hours. The reaction mixture is filtered to separate the catalyst and then concentrated in vacuo. The crude product is triturated with ether and filtered.

   The precipitate is taken up in water and lyophilized to obtain 470 mg of methyl ester monomethanesulfonate of (i) -1- [N- [3- (benzoyl-

  <Desc / Clms Page number 126>

 
 EMI126.1
 amino); melting point of 80-140 C; (c = 1.10; methanol). 0.63 (Be silica gel; chloroform / methanol / acetic acid; 9/1/1).



   Analysis: calculated for C26H31N305. CH403S
C: 56.98; H: 6.34; N: 7.30; S: 5.63
Found: C: 56.98; H: 6.32; N: 7.47; S: 5.63
Example 111.



   (-) - 1- [N- [3- (benzoyl-amino) -2-oxo-4-phenylbutyl] -L-alanyl] -L-proline methyl ester hemisulfate.



   The methyl ester is taken up in methanol (40 ml) with IN sulfuric acid (0.9 ml).
 EMI126.2
 than (i) butyl] -N- (600 mg; 1 mmol) prepared as described in Example 109 (e). Palladium on carbon catalyst (10%; 120 mg) is added and the reaction mixture is stirred under hydrogen 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 and lyophilized to obtain 405 mg of (¯) -1- [N- [3- (benzoylamino) - methyl ester hemisulfate) 2-oxo-4-phenylbutyl] -L-alanyl] -L-proline; melting point of 98? -
 EMI126.3
 112 C (850C); [a] 23 = (c = 1.16; methanol).



  D -60, 30 Rf 0.63 (spreading) (silica gel; chloroform / methanol / acetic acid; 9/1/1).



  Analysis: calculation for C26H31N3O5.0.5H2SO4.0.91H2O:
C: 58.81; H: 6.42; N: 7.92; S: 3.01 Found: C: 58.81; H: 6.11; N: 7.91; S: 3.09.

  <Desc / Clms Page number 127>

 



   Example 112.



  () -1- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -L-alanyl] -Lproline propyl ester monomethanesulfonate. a) Propyl ester of (-) - l- [N- [N- [3- (benzoylamino) - 2-oxo-4-phenylbutyl] -N- [(phenylmethoxy) carbonyl] -L- alanyl-L-proline.



   We use (-) - l-fN- [N- [3- (benzoyl-
 EMI127.1
 amino) L-alanyl] -L-proline (1.17 g; 2 mmol) prepared as described in Example 109 (d), in tetrahydrofuran (2 ml) with stirring in an ice bath.



  To this, n-propanol (3.0 ml; 40 mmol) is added followed by 4-dimethylaminopyridine (122 mg; 1 mmol) and dicyclohexylcarbodiimide (412 mg; 2 mmol).



  The reaction is allowed to warm to room temperature and continue overnight. 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 a mixture of benzene and acetic acid (9/1) to obtain 1.0 g of propyl ester of (-) - l-
 EMI127.2
 [N- [N- [3- nylmethoxy) carbonyl] -L-alanyl] -L-proline. b) Monomethanesulfonate of propyl ester of (-) - 1 - [N - [- 3-benzoylamino) (benzoylamino) -2-oxo-4-phenylbutyl] -N- [(phe-L-proline.



   The ester product from paragraph (a) is taken up (500 mg; 0.8 mmol) in methanol (50 ml). Methanesulfonic acid (0.72 ml of IN solution in methanol) and palladium catalyst are added.

  <Desc / Clms Page number 128>

 on carbon (10%; 100 mg) and the reaction mixture is stirred under hydrogen overnight. The reaction mixture is filtered to separate the catalyst, concentrated to dryness, triturated with ether and filtered to obtain 406 mg of crude product.

   This crude product is taken up in water, filtered through a millipore and lyophilized to obtain 390 mg
 EMI128.1
 (-) - 1- [N-t3-L-proline propyl ester monomethanesulfonate; melting point of 82 -94 C (69 C); [at] ; (c = 0.95; methanol). 0.46 D f (silica gel; chloroform / methanol / acetic acid; 90/5/5).
 EMI128.2
 o Jb J b 4 J z
C: 58.15; H: 6.73; N: 7.01; S: 5.34; Found: C: 58.15; H: 6.63; N: 7.05; S: 5.34.
 EMI128.3
 Example 113.



  Monomethanesulfate of ethyl ester of (-) - 1- {N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] -L-alanyl] -Lproline. a) Ethyl ester of (-) - l- [N- [N- [3- (benzoylamino) - 2-oxo-4-phenylbutyl] -N-f (phenylmethoxy) carbonyl] -L- alanyl] -L-proline.



   We use (-) - l- [N- [N- [3- (benzoyl-amino) -2-oxo-4-phenylbutyl] -N- [(phenylmethoxy) carbonyl] - L-alanyl] -L- proline (1.17 g; 2 mmol) prepared as described in Example 109 (d) in tetrahydrofuran (2 ml) with stirring in an ice bath. To this is added absolute ethanol (2.3 ml; 40 mmol) followed by 4-dimethylaminopyridine (122 mg; 1 mmol) and dicyclohexylcarbodiimide (412 mg; 2 mmol). Let the reaction continue

  <Desc / Clms Page number 129>

 overnight at room temperature. The dicyclohexylurea is filtered off, the filtrate is concentrated to dryness, the residue is taken up in ethyl acetate and washed until neutral with 10% potassium bisulfate and saturated sodium carbonate.

   The crude product (1.2 g) is purified on a column of silica gel in a mixture of benzene and acetic acid (8/2) to obtain 1.0 g of ester
 EMI129.1
 (-) - 1-phenylbutyl] -N- [N- [N- [3- (benzoylamino) -2-oxo-4-L-proline ethyl. b) Monomethanesulfate of ethyl ester of (-) - 1- [N- [3- (benzovlamino) -2-oxo-4-phenvlbutvl] -L-alanvl] -L- proline.



   The ester product from paragraph (a) is taken up (500 mg; 0.8147 mmol) in 50 ml of methanol and methanolic methanesulfonic acid (IN; 0.733 ml). 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,
 EMI129.2
 and lyophilized to obtain 360 mg of (-) - 1- [N-amino) -2-oxo-4-phenylbutyl] -L-alanyl] -L-proline ethyl ester monomethanesulfonate; melting point of 910-960C (720C); [a] 'D

  <Desc / Clms Page number 130>

 (c = 1.04; methanol). Rf 0.44 (silica gel; chloroform / methanol / acetic acid; 90/5/5). Analysis: calculationforC27H33N305.CH4SO. 0.6 H20
C: 57.33; H: 6.56; N: 7.17; 5: 5.47 Found: C: 57.33; H: 6.43; N: 7.12; S: 5.47.



   Example 114 Monomethanesulfonate of (¯) -1- [N- [3- (benzoylamino) -2-oxo-4-phenylbutyl] L-alanyl] -L-proline 1-methylethyl ester.
 EMI130.1
 a) L-methylethyl ester of (-) - l- [N-amino) [N- [3- (benzoyl-L-alanyl] -L-proline.



   We take again from (¯) -1- -l- [N- [N- [3-. (benzoyl-
 EMI130.2
 amino) L-alanyl] -L-proline (1.17 mg; 2 mmol) prepared as described in Example 109 (d) and isopropanol (3.0 ml; 40 mmol) in tetrahydrofuran (2 ml ) with stirring in an ice bath. Dicyclohexylcarbodiimide (412 mg; 2 mmol) is added to this.



  The reaction is allowed to continue overnight at room temperature. The reaction mixture is then concentrated to dryness, it is taken up in ethyl acetate and the dicyclohexylurea is separated by filtration. The filtrate is washed until neutral with 10% potassium bisulfate and saturated sodium bicarbonate. The crude product (1.2 g) is purified on a column of silica gel with a mixture of benzene and acetic acid (8/2) to obtain 1.0 g of l-methyl ethyl ester of (-) - l-
 EMI130.3
 [N- [N- [3- [b) 1-methylethyl ester monomethanesulfonate

  <Desc / Clms Page number 131>

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



   The ester product from paragraph (a) is taken up (815 mg; 1.3 mmol) 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 is concentrated to dryness in vacuo.



  The residue is triturated with ether and the precipitate is filtered to obtain 704 mg of crude product. This is taken up in 35 ml of water (2% solution), filtered through a millipore and lyophilized to obtain 600 mg of ester monomethanesulfonate 1-
 EMI131.1
 (-) - 1-4-phenylbutyl] -L-alanyl] -L-proline ethylmethyl; melting point --55, (c = 1.05; methanol).



  0.33 (silica gel; chloroform / methanol / [N- [3- (benzoylamino) -2-oxo-acetic acid; 9/1/1).



  Analysis: calculated for C28H35N3O5. CH4O3S. 0.66 H2O:
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 to 123.



   Following the method of Examples 109,112,
 EMI131.2
 113 and 114, (=) 2-oxo-4-phenylbutylJ-NT -1- [N- [3- (benzoylamino) -alanyl] -L-proline is treated with the reagent given below in Column 1 Separation of the alanyl protecting group gives the ester product given below in Column II.

  <Desc / Clms Page number 132>

 
 EMI132.1
 



  Collar. II
 EMI132.2
 
 EMI132.3
 Example Col. o 0 il 115 Ci-CH-0-C-C2H5-CH-0-C-C <0 0 U Il 116 Cl-CH-0-CC 1 CH (CH) 2 CH (CH) 0 0 il-Il 117 Cl-CH-OCC) -CH-OCC o 0 Il 118 Br-CH-0-C -CH-CH-0-C-CH

  <Desc / Clms Page number 133>

 
 EMI133.1
 Example Col. o 0 il 119 CI-CH-0-CO) -CH-O-C. 2 2 0 0 il 120 I-CH-COC) -CHC-0-C CH CH3 0 CH30 1 121 IC --- CO-CH-C - CO-CH 3 CH3 CH3 t 122) 2 22 OH OH 123 CH --- CH ---- CH-CH-CH-CH 1 OH 1 1 - <o) - <2)

  <Desc / Clms Page number 134>

 
 EMI134.1
 Example 124.



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



   Example 125
1000 tablets each containing the following ingredients are prepared: Sodium salt 1- [N - [(S) -3- (b enzoyl- amino) -2-oxo-4-phenylbutyl] -L- alanyl] -L-proline 100 mg.



  Corn starch 50 mg.



  Gelatin 7.5 mg.



  Avicel (microcrystalline cellulose) 25 mg.



  Magnesium stearate 2.5 mg. from sufficient bulk quantities in mixture
 EMI134.2
 managing the sodium salt of 1-2-oxo-4-phenylbutyl] -L-alanyl] -L-proline and corn starch with an aqueous gelatin solution. The mixture is dried and ground until a fine powder is obtained. We add the vicel and then the

  <Desc / Clms Page number 135>

 magnesium stearate until a granular mixture is obtained. This mixture is then compressed in a tablet press to form 100 tablets each containing 100 mg. active ingredient. Similarly, tablets containing 100 mg of the product of any of Examples 1 to 123 can be prepared.



   A similar process can be used to form tablets containing 50 mg of active ingredient.



   Example 126
We fill gelatin capsules n 1 made of two pieces each containing 50 mg of salt
 EMI135.1
 sodium of 1- [N- [butyl] -L-alanyl] -L-proline with a mixture of the following ingredients: Sodium salt of 1- [N- {(S) -3- (benzoylamino) -2 -oxo-4-phenyl-butyl] -L-alanyl] -L-proline 50 mg.



  Magnesium stearate 7 mg.



  Lactose 193 mq.



   250 mg.



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



   Example 127
A solution for injection is prepared as follows: Sodium salt of 1- [N- [(S) -3- (benzoylamino) -2-oxo-4-phenyl-butyl] -L-alanyl] -L-proline 500 g.



  Methyl paraben 5 g.



  Propyl paraben 1 g.

  <Desc / Clms Page number 136>

 



  Sodium chloride 25 g.



  Water for injection 5 g.



   The active substance, the preservatives and the sodium chloride are dissolved in
3 liters of water for injection and the volume is then brought to 5 liters. The solution is filtered through a sterile filter and poured under aseptic conditions into vials. previously sterilized which are closed by means of previously sterilized rubber closures. Each vial contains 5 ml of solution in a concentration of 100 mg of active ingredient per ml of solution for injection.



   Similarly, an injectable solution containing 100 mg of active ingredient per ml of solution can be prepared for the product of any of Examples 1 to 123.



   Example 128
1000 tablets each containing the following ingredients are prepared: Sodium salt of 1- [N- [(S) -3- (benzoyl- amino) -2-oxo-4-phenylbutyl] -L- alanyl] -L-proline 100 mg.



    Avicel 100 mg.



  Hydrochlorothiazide 12.5 mg.



  Lactose 113 mg.



  Corn starch 17.5 mg.



  Stearic acid 7 mg.



   350 mg. from sufficient bulk quantities by bringing in the form of agglomerates the sodium salt of

  <Desc / Clms Page number 137>

 
 EMI137.1
 1- [N- [alanyl] -L-proline, part of stearic acid. The agglomerates are ground and passed through a No. 2 sieve, then mixed with hydrochlorothiazide, lactose, corn starch and the remainder of stearic acid. The mixture is compressed into tablets in the form of 350 mg capsules in a tablet press. The tablets are marked to be divided in half.



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



   It should be understood that the present invention is in no way limited to the above embodiments and that many modifications can be made thereto without departing from the scope of this patent.


    

Claims (1)

 CLAIMS. 1. Composed of the formula:  EMI138.1  or a pharmaceutically acceptable salt thereof, wherein: X represents  EMI138.2    <Desc / Clms Page number 139>    EMI139.1  r - (L) - N-C-COOR .-, - N-C-COOR., - N, XL)) 6 H H H H or-N-CH-COOR R4 R4  EMI139.2  R represents hydrogen or a lower alkyl, cycloalkyl group,  EMI139.3    EMI139.4  - -NH2, - -0- - or-) -SH R-represents hydrogen or a lower alkyl group, lower alkyl substituted by halo,  EMI139.5    <Desc / Clms Page number 140>    EMI140.1  - "-) -NH-C" or- 3zo  EMI140.2  with the proviso that R represents hydrogen only in the case where R is different from hydrogen  EMI140.3  R represents - '' 2'0, - '2'J), (R14} - (CH2) rnto or - (C: 1z) m - &commat;. - (CH) M-ii-i or- (c, q) .-, 0 2 2 m, 9 0 N  EMI140.4  R hydrogen or a lower alkyl group,  EMI140.5    EMI140.6  - I.- OJ, substituted 2 by halo, - 2 "OH - 2r-TYo1'- -N, N I H H  EMI140.7  - (CH) -NH, -) -SH, -) -S-alkyl  <Desc / Clms Page number 141>    EMI141.1  NH 0 or or '' - NH-  EMI141.2  represents hydrogen or a lower alkyl group,  EMI141.3    EMI141.4  represents hydrogen or a  EMI141.5  lower alkyl, 2  EMI141.6    <Desc / Clms Page number 142>    EMI142.1    EMI142.2  -) -NH, -) -SH, -) -S-aLkyle lower,  EMI142.3  NH 0 II - (CH-NH-C) -C-NH 'NH-  EMI142.4  r is an integer from 1 to 4;  R represents from a lower alkyl group,  EMI142.5  halogen, c-to group, 0, R. q 0 / R19 Il hydroxy, -NH-C-aLkye férieurazido,, 'R20  EMI142.6    <Desc / Clms Page number 143>    EMI143.1  a 1-or 2-naphthyl group of the formula  EMI143.2  - (CH-), - O --Il Q - 2Cm * \ 0 '"15 3  EMI143.3  a 1-or 2-naphthyloxy group of the formula  EMI143.4  -), 'S'alkyle Lµ &commat; -' 4'p - (R13) 2. 0 or a group 1- (R 13) p  EMI143.5  of the formula  EMI143.6    <Desc / Clms Page number 144>    EMI144.1  Q R8 represents a group -15, a halogen, n 15 -) Q, -0-lower alkyl, a group 1-or n (Dl 1-- (R13)  EMI144.2  2-naphthyloxy of the formula  EMI144.3  -., - S-lower alkyl, 'ë &commat; - '' p -) - 0- or a group 1 - or 2- 2 m  EMI144.4  formula naphthylthio  EMI144.5    EMI144.6  represents a keto group or  EMI144.7    <Desc / Clms Page number 145>    EMI145.1  represents a halogen or -Y-R--; J-U 16 R-, R '-,' RetR 'are independently selected from hydrogen and lower alkyl radicals or else R' -, R and R'12 represent hydrogen and R represents  EMI145.2   R13 represents hydrogen, a lower alkyl group of 1 to 4 carbon atoms, a lower alkoxy group of 1 to 4 carbon atoms, a lower alkylthio group of 1 to 4 carbon atoms or a chloro, bromo, fluoro group , trifluoromethyl, hydroxy, phenyl, phenoxy, phenylthio or phenylmethyl; R14 represents hydrogen or a lower alkyl group of 1 to 4 carbon atoms, lower alkoxy of 1 to 4 carbon atoms, lower alkylthio of 1 to 4 carbon atoms, chloro, bromo, fluoro, trifluoromethyl or hydroxy; m is zero, one, two, three or four; p is equal to one, two or three provided that p is not greater than one except in the case where R13 or R14 represents hydrogen or a methyl, methoxy, chloro or fluoro group; R15 represents hydrogen or a lower alkyl group of 1 to 4 carbon atoms; Y represents oxygen or sulfur;    EMI145.3  R16 lower alkyl group J-b  <Desc / Clms Page number 146>  from 1 to 4 carbon atoms or a group  EMI146.1    or else the R groups join to form an unsubstituted pentagonal or hexagonal ring, this ring possibly comprising a lower alkyl substituent of 1 to 4 carbon atoms or a di (lower alkyl substituent of 1 to 4 carbon atoms) on one or more of its carbon atoms; R19 represents a lower alkyl, benzyl or phenylmethyl group; R20 represents hydrogen or a lower alkyl, benzyl or phenetyl group; R6 represents hydrogen or a lower alkyl, benzyl, benzhydryl group,  EMI146.2   R17 represents hydrogen or a lower alkyl, cycloalkyl or phenyl group;    EMI146.3  hydrogen or a group - Lo lower alkyl, lower alkoxy or phenyl, or R'7 taken together represent a group 17 IR--  EMI146.4  - (CI-12) -, '23' '= /  <Desc / Clms Page number 147>    EMI147.1  represents hydrogen or a  EMI147.2  lower alkyl group, - g, S 0 N  EMI147.3  are independently selected from hydrogen and lower alkyl groups; and represents a lower alkyl group 2.  Compound according to Claim 1, characterized in that: R represents hydrogen or a lower straight or branched chain alkyl or phenyl group R represents hydrogen or a lower straight or branched chain alkyl group of 1 to 4 atoms carbon, CF, - (CH) -NH,  EMI147.4  - - O) -OH, OH 'TTO)' '2-r'-CH.-SH, 2' 1 HH - (CH) -S-CH, - -OH, NH2 0 0 or - CH - C- NH--) -C-NH  <Desc / Clms Page number 148>    EMI148.1  with the proviso that hydrogen only represents in the case where R is different from hydrogen R of hydrogen or a cyclohexyl or phenyl group; R of hydrogen or a lower straight or branched chain alkyl group from 1 to  EMI148.2  4 ouzo carbon atoms. - CH- / 0 \ -OH, -CHO) -OH, -CH-OH, OH -CH N '2 H H 1 &num; NH - NH 0 0 Il -  EMI148.3  R represents hydrogen, a lower alkyl group with a straight or branched chain of 1 to 4 carbon atoms, an ion of an alkali metal salt,  EMI148.4    <Desc / Clms Page number 149>   R18 represents R17 represents hydrogen or a lower straight or branched chain alkyl group of 1 to 4 carbon atoms or cyclohexyl; R18 represents a straight or branched chain lower alkyl group of 1 to 4 carbon atoms or phenyl; R-represents a chain alkyl group  EMI149.1  straight or branched from 1 to 4 carbon atoms  EMI149.2  R - m-- 14  EMI149.3  R3 is a lower straight or branched chain alkyl group of 1 to 4 atoms  EMI149.4  carbon, (CH) -NH-, / r or- R14 "N N;  R7 represents hydrogen or a hydroxy group, lower straight or branched chain alkyl of 1 to 4 carbon atoms, cyclohexyl, amino, -O- lower alkyl in which the lower alkyl is a straight or branched chain of 1 to 4 carbon atoms,  EMI149.5  l-naphthyloxy, 2-naphthyloxy, -S-lower alkyl in which the lower alkyl is a straight or branched chain of 1 to 4 carbon atoms,  <Desc / Clms Page number 150>    EMI150.1    EMI150.2  - or 2-napntyltnio represents a group-O-lower alkyl L-naphthyltniorieur, -S-lower alkyl,  EMI150.3  in which the lower alkyl is a straight or branched chain of 1 to 4 carbon atoms, R represents a phenyl, 2-hydroxyphenyl or 4-hydroxyphenyl group;  both R10s represent a fluoro group,  EMI150.4  chloro or-Y-R--; 16 Y represents 0 or S represents a lower alkyl group - Lb with a straight or branched chain of 1 to 4 carbon atoms or else the R16 groups join to form an unsubstituted pentagonal or hexagonal ring, this cycle possibly comprising a methyl or dimethyl substituent on one or more of its available carbons;  EMI150.5  R-, R'i-j'PietR'represent all of hydrogen or else R11 represents a phenyl group, 2-hydroxyphenyl or 4-hydroxyphenyl and R ', R and R'12 all represent hydrogen; r is an integer from 1 to 4; m is zero, one or two; R13 represents hydrogen or a methyl, methoxy, methylthio, chloro, bromo, fluoro or hydroxy group;    <Desc / Clms Page number 151>   R14 represents hydrogen or a methyl, methoxy, methylthio, chloro, bromo, fluoro or hydroxy group; and R24 represents a phenyl group.  3. Compound according to claim 2, characterized in that: X represents a group  EMI151.1    EMI151.2  or or Y COOR6  EMI151.3  R represents from. hydrogen or methyl; R represents hydrogen, methyl or a group - (CH2) 4-NH2, with the proviso that Rlne represents hydrogen only in the case where R is different from hydrogen; R4 represents a cyclohexyl group or  <Desc / Clms Page number 152>    EMI152.1  phenyl R6 of hydrogen, a lower alkyl group with a straight or branched chain of 1 to 4 carbon atoms or an ion of an alkali metal salt; R7 represents hydrogen or a cyclohexyl or lower alkoxy group of 1 to 4 atoms  EMI152.2  carbon, - 'R13 - O-H,),) O). 13 13  EMI152.3  m is zero, or two (CH) IlR13 represents hydrogen or a methyl, methoxy, methylthio, chloro, bromo, fluoro or hydroxy group; and t is 2 or 3.  4. Compound according to claim 3, characterized in that X represents a group  EMI152.4   5. Compound according to claim 4, characterized in that R represents a phenyl group and R3 a benzyl group.  6. Compound according to claim 5, characterized in that R represents hydrogen, RI of methyl, R, of cyclohexyl and R6 of hydrogen.  7. Compound according to claim 5, characterized in that R represents hydrogen,  <Desc / Clms Page number 153>    EMI153.1  R. methyl, phenyl and hydrogen. D-4 b 8. Compound according to claim 5, characterized in that R represents methyl, R  EMI153.2  hydrogen, cyclohexyl and hydrogen. 4 o 9. Compound according to claim 3, characterized in that X represents a group  EMI153.3   10. Compound according to claim 9, characterized in that R2 represents phenyl and R3 represents benzyl.  11. Compound according to claim 10, characterized in that R represents methyl, R of hydrogen and R6 of hydrogen.  12. Compound according to claim 10, characterized in that R represents methyl, R of hydrogen and R6 of methyl.  13. Compound according to claim 10, characterized in that R represents hydrogen,  EMI153.4  methyl and R6 hydrogen.  14. Compound according to claim 3, characterized in that X represents a group  <Desc / Clms Page number 154>    EMI154.1   15. Compound according to claim 14, characterized in that R2 represents phenyl and R3 of benzyl.  16. Compound according to claim 15, characterized in that R represents methyl, R of hydrogen and R6 of hydrogen.  17. Compound according to claim 15, characterized in that R represents hydrogen,  EMI154.2  R. methyl and R6 hydrogen. D-6 18. Compound according to claim 3, characterized in that X represents a group  EMI154.3   19. Compound according to claim 18, characterized in that R7 represents hydrogen.  20. Compound according to claim 19,  EMI154.4  characterized in that represents phenyl, du- (CH) -NH, hydrogen, and hydrogen.  R221. Compound according to Claim 19, characterized in that R2 represents phenyl, R3 from (CH) ..- CH, R from hydrogen, R from methyl and R6 from hydrogen.  <Desc / Clms Page number 155>    EMI155.1   22. Compound according to claim 19, characterized in that R2 represents phenyl,  EMI155.2  R Jan R3  EMI155.3  R of hydrogen, R of methyl and hydrogen. 23. Compound according to claim 19, characterized in that R2 represents phenyl,  EMI155.4  R represents -CH 2  EMI155.5  R hydrogen, du and hydrogen.  24. Compound according to claim 19, characterized in that R2 represents phenyl and  EMI155.6  benzyl.  25. Compound according to claim 24, characterized in that R represents hydrogen, R of methyl and R6 of hydrogen.  26. Compound according to claim 25, characterized in that it consists of 1- [N- [(S) -3- (benzoylamino) -2-oxo-4-phenylbutyl] -L-alanyl] monohydrochloride - L-proline.  27. Compound according to claim 24, characterized in that R represents hydrogen, R1 of methyl and R6 of methyl.  28. Compound according to claim 24, characterized in that R represents hydro-  EMI155.7  gene, R of methyl and ethyl. i 6 29. Compound according to claim 24, characterized in that R represents hydrogen, R of methyl and R6 of n-propyl.  <Desc / Clms Page number 156>    30. Compound according to claim 24, characterized in that R represents hydrogen,  EMI156.1  R of methyl and isopropyl. - L b 31. Compound according to claim 24, characterized in that R represents methyl, R  EMI156.2  hydrogen and hydrogen. 6 32. Compound according to claim 24, characterized in that R represents hydrogen, R1 a group - (CH2) 4-NH2 and R6 hydrogen.  33. Compound according to claim 18, characterized in that R7 represents a group  EMI156.3   34. Compound according to claim 33, characterized in that R2 represents phenyl and R3 of benzyl.  35. Compound according to claim 34, characterized in that R represents methyl, R  EMI156.4  hydrogen and hydrogen. 6 36. Compound according to claim 18, characterized in that R7 represents a group  EMI156.5   37. Compound according to claim 36, characterized in that R2 represents phenyl and R3 represents benzyl.  38. Compound according to claim 37, characterized in that R represents methyl, R  EMI156.6  hydrogen and R6 hydrogen. b  <Desc / Clms Page number 157>   39. Compound according to claim 37, characterized in that R represents methyl,  EMI157.1  Ru hydrogen and methyl. 40. Compound according to claim 37, characterized in that R represents hydrogen, R of methyl and hydrogen. de41. Compound according to Claim 18, characterized in that R7 represents phenyl.  42. Compound according to claim 41, characterized in that R2 represents phenyl and R3 of benzyl.  43. Compounds according to claim 42, characterized in that R represents hydrogen, Ru of methyl and R6 of hydrogen.  44. Compound according to claim 18, characterized in that R7 represents cyclohexyl.  45. Compound according to claim 44, characterized in that R2 represents phenyl and R3 represents benzyl.  46. Compounds according to claim 45, characterized in that R represents hydrogen,  EMI157.2  R of methyl and hydrogen. 47. Compound according to claim 1, characterized in that R represents a lower alkyl, cycloalkyl group,  EMI157.3  - (CH2) '-  EMI157.4  - (CH,), - NH ,, - -OH.- 2 4 - (CH,), - OH, - or 2 4 2- -) -SH, ..  <Desc / Clms Page number 158>    EMI158.1  and in that m is zero, one, two, three or four. 48. Compound according to claim 47, characterized in that R represents methyl.  EMI158.2   49. Compound of the formula: 0 R 0 Il R3-CH-C-CH2-NH-CH-C-X 1 (L) NH I! 1 R2  EMI158.3  or a pharmaceutically acceptable salt thereof, formula in which: X represents:  EMI158.4    <Desc / Clms Page number 159>    EMI159.1    EMI159.2  R represents a lower group, lower alkyl substituted by halo,  EMI159.3    <Desc / Clms Page number 160>    EMI160.1  - 2) N):  OD- (CH2) r'2 r N Ij 1 HHH - (CH-) -NH, -) -SH, -) -S-alkyl NH - (CH) -NH-C 2 "NH NH2 - - (CH2) m 0 -2nr (14) N n N R3 represents hydrogen, a lower alkyl group,  EMI160.2    <Desc / Clms Page number 161>    EMI161.1  lower alkyl. substituted killed! o, - (CH.,) - cycloalkvLe,  EMI161.2  - OY-OH, -, - OH 2 2 r 2 OH H 'rT'r' .-) -NH, -) -SH, N N NH NH - (CH) -S-aIkyleinSrLeu- NH2 0 il -  EMI161.3  R5 represents hydrogen, a lower alkyl group, 5  EMI161.4    <Desc / Clms Page number 162>    EMI162.1  - (CH2) H "N H H H H - (CH) -NH, -) -SH, -) -S-loweralkyl, 0 2 - f NH 0 - H-C- - NH2  EMI162.2  r is an integer from 1 to 4 represents from a lower alkyl, halogen, keto group,  EMI162.3  o / R19 il-N hydroxy, -NH-C-alkyl o 'azidoami.-, io 20 "-R20 Il - -, - <2rr ri 2 m (R14) p (R13) p - 2)' -2)  '- (CH2m- o N zo 5  <Desc / Clms Page number 163>    EMI163.1  a 1-or 2-naphthyl group of the formula  EMI163.2  - (CH) -cycloalkyle, - (CH 2) o (R14) p 2 m "'"> <]. . S &commat; - '' p 0 - -N ".-0-aiad.) 0 \, 15 3" * R15 M 0 (Pl 3) p  EMI163.3  a 1-or 2-naphthyloxy group of formula r  EMI163.4  -0- lower, m o (R 14) p - group 1- hio m (Rm3 \ 13'p  EMI163.5  of the formula  EMI163.6    <Desc / Clms Page number 164>    EMI164.1  a R8 represents a keto group, 8 il 11-11 15 a halogen, -o-C-N "" 15 -) -0-.). lower, a group 1- (13)  EMI164.2  or 2-naphthyloxy of the formula  EMI164.3  - -S-lower alkyl, Ig 'p - - a group 1-or 2- 2 13 P  EMI164.4  formula naphthylthio  EMI164.5    EMI164.6  Rn represents a keto group or  EMI164.7    <Desc / Clms Page number 165>    EMI165.1  R represents a halogen or-Y-R. ;  10 l R, R '-, i'R - .., stR're independently of hydrogen or a lower alkyl group, represent hydrogen and Rll represents a group  EMI165.2   R13 represents hydrogen or a lower alkyl group of 1 to 4 carbon atoms, lower alkoxy of 1 to 4 carbon atoms, lower alkylthio of 1 to 4 carbon atoms, chloro, bromo, fluoro, trifluoromethyl, hydroxy , phenyl, phenoxy, phenylthio or phenylmethyl; R14 represents hydrogen or a lower alkyl group of 1 to 4 carbon atoms, lower alkoxy of 1 to 4 carbon atoms, lower alkylthio of 1 to 4 carbon atoms, chloro, bromo, fluoro, trifluoromethyl or hydroxy ; m is zero, one, two, three or four;  p is equal to one, two or three, with the proviso that p is greater than 1 only in the case where R13 or R14 represents hydrogen or a methyl, methoxy, chloro or fluoro group; R15 represents hydrogen or a lower alkyl group of 1 to 4 carbon atoms; Y represents oxygen or sulfur;  EMI165.3  R16 represents a lower alkyl group 16  <Desc / Clms Page number 166>    EMI166.1  from 1 to 4 carbon atoms or a group  EMI166.2    EMI166.3  or else the R16 groups join to form an unsubstituted pentagonal or hexagonal ring Jb, this cycle possibly comprising a lower alkyl substituent of 1 to 4 carbon atoms or a di substituent (lower alkyl of 1 to 4 carbon atoms) on one or several of its carbon atoms;     R19 represents a lower alkyl, benzyl or phenetyl group; R20 represents hydrogen or a lower alkylbenzyl or phenetyl group; and  EMI166.4  hydrogen or a group 6 lower alkyl, benzyl, benzhydryl or  EMI166.5  in which R17 represents hydrogen or a group  EMI166.6  lower alkyl, cycloalkyl or phenyl, and J-O has hydrogen or a lower alkyl, lower alkoxy or phenyl group, or and J- / JLo taken together represent a group  EMI166.7    <Desc / Clms Page number 167>   50.  Pharmaceutical composition used for the treatment of hypertension comprising a pharmaceutically acceptable carrier and a hypotensive compound of the formula:  EMI167.1  wherein R, R1, R2, R3 and X are as defined in claim 1.  51. A method of treating hypertension in a mammalian host, characterized in that it comprises the administration of an effective amount of the composition of claim 50.  52. Pharmaceutical composition which can be used as an analgesic comprising a pharmaceutically acceptable carrier and an enkepininase inhibitor compound of the formula:  EMI167.2    EMI167.3  in which R, R., R., R.R and are such as J- j b 6 defined in claim 1.  53. A method of relieving pain in a mammalian host, characterized in that it comprises the administration of an effective amount of the composition of claim 52.  <Desc / Clms Page number 168>    EMI168.1  54. Compound of the formula  EMI168.2    EMI168.3  in which halo represents Cl or Br represents a group  EMI168.4    EMI168.5  R3 represents hydrogen or a'- (CH2) 'lower alkyl group, 4p  EMI168.6    EMI168.7  - (-J 'lower substituted by halo, -) -cycloalky, -) -0) -OH r OH - (CH'- -OH, H 1 H n  <Desc / Clms Page number 169>    EMI169.1  - 2 2 r 21 2 r 1 H H NH H - (CH) -S-aDsiisoeor, '2 r 2 r N, NH2 0 Il - (CH) -C-NH-  EMI169.2  represents hydrogyne or a lower alkyl group of 1 to 4 carbon atoms, lower alkoxy of 1 to 4 carbon atoms, alkylthio of less than 4 carbon atoms, chloro,  bromo, fluoro, trifluoromethyl or hydroxy m is zero, one, two, three or four p is equal to one, two or three, provided that p is not greater than one except in the case of hydrogen or a methyl, methoxy, chloro or fluoro group and r is an integer from 1 to 4. 55. Compound according to claim 54, characterized in that R is a group  EMI169.3    <Desc / Clms Page number 170>     ; RI, is a group  EMI170.1  m is zero, one or two, and R14 represents hydrogen or a methyl, methoxy, methylthio, chloro, bromo, fluoro or hydroxy group.  56. Compound according to claim 55, characterized in that R represents  EMI170.2    c 3 represents  EMI170.3  and halo represents Cl.  EMI170.4   57. Compound of the formula: o 0 R R il R2-C-i-TH-C-CH2- CH-C-OH 1 Ri  EMI170.5  in which: R represents hydrogen, Prot or a lower alkyl, cycloalkyl group,  EMI170.6  H -) -Q, -) -N-Prot, -) -N-Prot, m H - (CH2)  EMI170.7  - (CH) -0-Prot- (CH) - S-Prot, - (CH) -S-Prot or-) -S-Prot  <Desc / Clms Page number 171>    EMI171.1  represents hydrogen or a lower alkyl group, lower alkyl substituted by halo,  EMI171.2  - o-Prot, 2) r 0 (CH2) - (CHO-O-Prot.-) .--. j, 2 r 'O-Prot H - (CH N- (CH-S-Prot- (CH -) -r - N, -)) lower, N 1 Prot - N o or- the condition that R does not represent  EMI171.3  hydrogen than in the case where R is different from hydrogen;  Prot is an easily separable protection group R2 represents a group  EMI171.4    <Desc / Clms Page number 172>    EMI172.1  <1 represents hydrogen or a lower alkyl group,  EMI172.2    EMI172.3  - (CH2) - OC, lower alkyl substituted by halo, - (CH) -cycloalkyle, -) --0) --- OH r OH - (CH2) '- 2) HH - -) -NH-) - SH, N r. H. - (CH) -S-alle  EMI172.4    <Desc / Clms Page number 173>    EMI173.1  represents. or a lower alkyl radical of 1 to 4 carbon atoms, lower alkoxy of 1 to 4 carbon atoms, lower alkylthio of 1 to 4 carbon atoms, chloro, bromo, fluoro, trifluoromethyl or hydroxy; m is equal to zero, one, two, three or four p is equal to one, two or three, on the condition that p is greater than one only in the case where represents hydrogen or a methyl or methoxy group, chloro or fluoro; and r is an integer from 1 to 4. 58. Compound according to claim 57, characterized in that R2 represents a group  EMI173.2   0 R3 represents a group  EMI173.3  m is zero, one or two; R14 represents hydrogen or a methyl, methoxy, methylthio, chloro, bromo, fluoro or hydroxy group; R represents a methyl group; and  EMI173.4  R represents hydrogen.  59. Compound according to claim 58, characterized in that R2 represents phenyl and R3 represents benzyl.  <Desc / Clms Page number 174>    60. Compound according to claim 57, characterized in that: R represents  EMI174.1   R3 represents  EMI174.2  m is 0, 1 or 2; R14 represents hydrogen or a methylgmethoxy, methylthio, chloro, bromo, fluoro or hydroxy group; R represents an easily separable protection group, and R represents methyl.  61. Compound according to claim 50, characterized in that R2 represents phenyl, R of  EMI174.3  benzyl and R Il - -0 /.  EMI174.4   62. Substituted peptide compounds and their intermediates, their preparation and their use, as described above, in particular in the Examples given.