BG60339B2 - Derivatives of amino acids as antihypertensive agents - Google Patents

Abstract

The new compounds are used in medicine as antihypertensive agents. A group of new carboxyalkyl dipeptides and their derivatives were obtained, pharmaceutical compositions and methods for the treatment of high blood pressure were created. 82 claims

Description

Substituted acyl derivatives of amino acids which are useful as inhibitors of the angiotensin converting enzyme (1) and (2), in particular, these compounds are mercapto substituted acyl amino acids and their derivatives include the clinically effective anti-hypertensive compound-captopril 2-methylpropanoyl-1-proline.

Compounds known in the art are not dipeptide derivatives, such as the compounds of the invention. Compounds known in the art contain, as an essential substituent, sulfhydryl or a derivative thereof, while those of the present invention do not contain one. In addition, the dipeptide compounds of the invention are dipeptides whose N-terminals bear a carboxymethyl group, which is preferably further substituted by a methyl group. The carboxyl group (s) may also be converted to an ester, an amide or a salt. In fact, the compounds of the invention are hybrids formed by combining .beta.-amino acids with dipeptides using nitrogen, which is common to these two partial structures. Such structural arrangement is uncommon in the field of synthetic and natural peptides and is not described in the mercaptoacyl type of functions in the above two patents of the prior art.

Bj-carboxyalkanoyl amino acids are known which are useful as inhibitors of the antipiotensin converting enzyme (3). Because the compounds of the invention are dipeptide derivatives, in the formal sense, they can be considered to be related to some of the compounds described in patent lit. source / 3 /. When one of the methylene groups is substituted for an amino function, a high activity of the compounds is observed according to the invention. For example, the preferred compounds of the invention may be administered in solid low doses of 2.5 mg per day per patient against the lowest doses in the order of 1 mg per kilogram weight per day of the most preferred compounds described in lit. source / h / _t which corresponds to about 60 mg per day per patient based on the average patient weight - about 150 pounds.

SUMMARY OF THE INVENTION

The present invention relates to carboxyl dipeptides and derivatives thereof, which are administered as inhibitors of the converted enzyme and as antihypertensive agents. The compounds of the invention may be included in the following formula

0 II ? 1 * with 1 ^{J R} 5 0 And R - C - with - 1 NHCH · - p II - N -C - 1 C - Rg * 2 0 * 7

wherein

R and Rg are the same or different and denote hydroxy, lower alkoxy, lower alkenyloxy, dinisalkylamino lower alkoxy (dimethylaminoethoxy), acylamino lower alkoxy (acetylaminoethoxy), acyloxy lower alkoxy (pivaloyloxymethoxy), acryloxybenzoxy, acryloxybenzoxy, acryloxysiloxy aryloxy or substituted arylalkylalkoxy, wherein the substituent is methyl, halogen or methoxy, amino, lower alkylamino, dinisalkylamino, hydroxyamino, aryl lower alkylamino, such as benzylamine;

RJ means hydrogen, alkyl containing from 1 to 20 carbon atoms, which may have a branched or cyclic chain and includes unsaturated / such as allyl / alkyl groups, substituted lower alkyl, wherein the substituent may be halogen, hydroxy, lower alkoxy, aryloxy, for example phenoxy, amino, dinisalkylamino, acylamino, for example acetamido and benzamido, arylamino, guanidino, imidazolyl, indolyl, mercapto, lower alkylthio, arylthio, for example phenylthio, carboxy, or carboxamido, carboxyl, carboxyl, carboxyl like a fairy yl, wherein the substituent may be lower alkyl, lower alkoxy or halogen, aryl lower alkyl, aryl lower alkynyl, heteroaryl lower alkyl or heteroaryl lower alkenyl, such as benzyl, styryl or indolylethyl, substituted aryl alkyl, substituted alkylaryl substituted alkyl lower alkenyl, wherein the substituent (s) may (may) be halogen, dihalogen, lower alkyl, hydroxy, lower alkoxy, amino, aminomethyl, acylamino (acetylamino or benzoylamino), dinisalkyl amino, lower carbonyloxyalkyl , a cyano or sulfonamino group, aryl lower alkyl, heteroaryl alkyl, substituted alkyl moiety by amino or acylamino (acetylamino or benzoylamino);

R? and R? are the same or different and denote hydrogen or lower alkyl;

K ₃ represents hydrogen, lower alkyl, phenyl lower alkyl, aminomethylphenyl lower alkyl, hydroxy lower alkyl, acylamino lower alkyl /, such as benzoylamine lower alkyl, acetylamino lower alkyl, acetylamino lower alkyl, alkylsilyl, alkylsilyl di-alkylsilyl di-alkylsilylalkyl, lower alkyl, imidazolyl lower alkyl, indolyl lower alkyl, mercapto lower alkyl, lower alkylthio lower alkyl;

means hydrogen or lower alkyl;

R4 represents hydrogen, lower alkyl, phenyl, phenyl lower alkyl, hydroxy phenyl lower alkyl, hydroxy lower alkyl, amino lower alkyl, guanidinone lower alkyl, imidazolyl lower alkyl, indolyl lower alkyl, mercapto alkyl, mercapto alkyl, mercapto alkyl, mercapto alkyl, mercapto alkyl;

^R 4 and R ₅ ^{may be a} xylene bridge, bonded together and form an alloy containing from 2 to 4 carbon atoms, an alkylene bridge containing from 2 to 3 carbon atoms and one sulfur atom, an alkylene bridge containing 3 or 4 carbon atoms containing a double bond or an alkylene bridge substituted as above with hydroxy, lower alkoxy, lower alkyl or dinisalkyl; and their pharmaceutically acceptable salts.

The lower alkyl or lower alkenyl group, unless otherwise indicated, represents some of the many including straight or branched chain hydrocarbon radicals containing from 1 to 6 carbon atoms, for example methyl, ethyl, propyl, isopropyl, tert-butyl, pentyl, isopentyl, hexyl or vinyl, allyl, butenyl and the like. The aralkyl groups are clarified with each of the above, which contains from 1 to 4 carbon atoms and in the alkyl moiety and include, for example, benzyl, p-methoxy benzyl and the like. Halogen means chlorine, bromine, iodine or fluorine. The term aryl, where it appears, unless otherwise indicated, means phenyl or naphthyl. Heteroaryl groups include, for example, pyridyl, thienyl, indolyl, benzthienyl, furyl, imidazoyl and thiazolyl.

The R 1, R ₃ and R ₅ substituted lower alkyl moieties are explained by the following groups of formulas

H0-CH ₂ -, H5-CH ₂ - ^{H, H} ₂ N- / CH _2/4 -, * CH ₃ -5 / CH _2/2 -, H ₂ N- / CH _2/3 -,

NH

H ₂ NC -NH- / CH _2/3 - and others.

When R ₄ and R 4 are bonded via a carbon atom and the nitrogen atom to which they are attached, they form a four to six membered ring which may contain a sulfur atom or a double bond. Preferred rings have the following formulas in which Y represents a methylene group, a sulfur atom or CHOCH ₃ . Preferred are those compounds of the general formula (I) in which R and Rg may independently represent hydroxy, lower alkoxy, lower alkenyloxy, arylisalkoxy, amino, dinisyl alkylamino lower alkoxy, acylamino lower alkoxy, ocyloxy lower alkoxy, ocycloxy lower is methyl, halo or m methoxy;

R ₂ and denote hydrogen;

1? ₃ means lower alkyl, amino lower alkyl, imidazoyl lower alkyl, halo lower alkyl;

< 1 > are linked and preferably form rings such as those mentioned above in which Y represents CH ₂ , sulfur or

CH-OCH ₃ ; 1C has the meanings given above.

Even more preferred are the compounds of the general formula (I) in which R <1 means alkyl containing from one to eight carbon atoms, substituted lower alkyl, wherein the alkyl group contains one to five carbon atoms and the substituent is amino, alkylthio, arylthio, aryloxy or arylamino, arylalkyl or heteroarylalkyl, the alkyl portion having one to three carbon atoms atoms such as phenylethyl or indolylethyl or substituted aryl lower alkali (phenyl lower alkyl or continuous lower alkyl) and substituted heteroaryl alkyl, the alkyl group having one to three carbon atoms and the substituent (s) being halogen, two halogen atoms, two halogen atoms, alkyl, hydroxy, lower alkoxy or lower alkyl. The most preferred compounds of the general formula (I) are those in which

R and Rg are hydroxy, lower alkoxy, aralkyloxy;

R ₂ and R ₇ denote hydrogen;

R ₃ represents methyl or amino lower alkyl;

R | and Κς are bonded via carbon and nitrogen to the formation of proline, 4-thiaproline or 4-methoxyproline;

R4 represents alkyl containing from 1 to 8 carbon atoms, substituted lower alkyl, the alkyl group containing one to five carbon atoms and the substituent being amino, arylthio or aryloxy, arylalkyl or heteroarylalkyl, the alkali moiety containing one to three carbon atoms, such such as phenethyl or indolylethyl or substituted arylalkyl (phenyl) lower alkyl or naphthyl lower alkyl) and substituted heteroarylalkyl, the alkyl substituent having one to three carbon atoms and being substituted by one or two substituents such as halogen, dihalogen, amino, hydroxy, hydroxy, amino, hydroxyissh alkoxy or lower alkyl.

Preferred, particularly preferred and most preferred compounds also include their pharmaceutically acceptable salts and are explained by the following compounds: N- (1- (5) -carboxy-3-phenylpropyl) -L-alanyl-L-proline; N- (S) -ethoxycarbonyl-3-phenylpropyl] -L-alanyl-L-proline and their salts;

N- (N) -5-ethoxycarbonyl-4-methylpentyl) -L-alanyl-L-proline; L1- (1-carboxy-5-aminopentyl) -L-alanyl-L-proline;

- (N) 5-Carboxy-3-phenylpropyl] -L-lysyl-L-proline;

N-Z, (1-5) -ethoxycarbokinyl-3-phenylpropyl) -L-proline; N- [1- (5) -carboxy-3- (3-indolyl) propyl] -L-lysyl-L-proline; N-Z (1/5) -carboxy-3- (4-chlorophenyl) -propyl] -L-lysyl-L proline;

N-Z (1/6) -carboxy-2-phenylthioethyl) -L-lysyl-L-proline; N- [N (S) -carboxy-3- (4-chlorophenyl) -propyl] -L-lysyl-1- (4) -methoxypropyl;

N-N (1/5) -carboxy-5-aminopentyl-L-lysyl-L-proline: Ethyl N- (N) -5-ethoxycarbonyl (-3-phenylpropyl) -L-alanyl-L-proline hydrochloride and the like.

The compounds of formula (I) and preferred subgroups may be prepared by one or more methods selected from the ones described below.

0 and ^R 1 ^R 3 0 II % ^R 5 0 p R - C -  With -NHC - 1 C - • N - C - 1 C - R ₆ r ₂ ^ 7

As shown in the examples, reactive groups which are not included in the conjugations such as amino, carboxy, mercapto, etc. may be protected in advance by some of the standard methods used in peptide chemistry before the addition reactions and then released until the desired products are received.

Method I. Method I / R ₂ means H /

OR, R_ 0 R _£ Re 0

I ¹ I ³ 11 I * I ⁵ ll

R - C - C = 0 ♦ H ₂ NC -CNCC-Rg NaBHgCHH / I / ^R 7 / II / / III /

The keto acid (or ester, amide or hydroxamic acid) (II) is condensed with dipeptide (III) in an aqueous solution, optimally under conditions close to neutral, or in a suitable organic solvent (eg CH ₃ SK) in the presence of sodium cyanoborohydride to give of / I / R ₂ means H /. Optionally, the intermediate Schiff base, enamine or aminol can be catalytically reduced to a compound of formula (I) using, for example, hydrogen in the presence of 10% palladium-on-carbon or Raney-nickel. The ratio of the resulting diastereomeric compounds can be varied by the choice of catalyst.

If R and R ₆ are protected carboxy groups such as alkoxy or benzyloxy or the like, they can be converted by known methods such as hydrolysis or hydrogenation to a product of formula (I) in which I? and / or R8 are hydroxy. this also applies to all subsequent methods in which such a situation exists.

In one case, a compound of formula (II) may condense with an amino acid of formula (IU) /% 0 R 1 R ³ - μ 1 ¹ 1

H ₂ NCH-C00H * II PAV ₃ SC R - C -CHNCHCOH (I // Y) under the same conditions to obtain an amino acid Ϊ. Subsequent accession by known methods to an amino acid derivative of formula ΪΙ results in a compound of formula (I).

The known methods provide reactive groups with protection during addition reactions, for example with N-formyl, N-tert-butoxycarbonyl and N-carbobenzyloxy groups, followed by their removal to obtain product I. Moreover, the substituent R may include ester removable groups such as benzyl, ethyl or tert-butyl. The condensing agents in this synthetic pathway are commonly used in peptide chemistry, such as dicyclohexylcarbodiimide (DSS) or diphinylphosphoryl ozide (DPPA) or a compound of formula Y can be activated by intermediate active esters similar to those derived from 1-hydroxybenzotri.

/ VAT / / I / * 7 / ¥ 1 / / VAT / = / DPP / =

Dicyclohexylcarbodiimide Diphenylphosphoryl azide

Method / ¥ P / / ¥ W /

An amino acid / or ester, amide or hydroxamic acid / ¥11 is reacted with ketone ¥ 111 under the conditions described above.

Method I for Preparation of Compound of Formula I.

In one case, the synthesis can be carried out in stages by condensation of a compound of formula ¥ 11 with keto acid of formula IX

R, 0 R.R

I II I

YII * 0 = C - COOH. R-C - C - NH - CH - COOH

---------------> I ^R 2 / IX // X / to obtain the amino acid of formula X. By known methods, as indicated above in Experiment (method / 1), a compound of formula X may be attached to an amino acid derivative of formula ¥ 1 to obtain a compound of formula I.

HN / P />

n-I / I / when carrying the J1-amino substituent, the carbonyl and amino groups can be protected in the form of β-Lactam function ·

Model

2. Method I ^R 1 * 1 ^R 3 ^{0 R} 4 ^R S

I ') il <I

-C-COR - * ROCCNHCH-CNC-CORg ^R ?

ι

R ₂

I

R ₂ (I) (XI) iodine or alkyl sulfonyloxy and aryl sulfo (III)

X stands for chlorine, bromine, nyloxy. The dipeptide of formula III is alkylated with a suitable X haloacid (in the form of an ester or amide) or with a? Z-sulfonyloxy acid (ester or amide) of formula XI under alkaline conditions in water or in an organic solvent.

In one case, the synthesis can be carried out in stages. ^R 3 /

H ₂ K1CH-C00H / I / / P /

I ¹ | ³ I ⁵ Ri R, 0 Ri Rc I ¹ i ³ // I ⁴ P RC0-C-NH-CH-C0H + HN- 1 -C-COR _g ——> RCO-C-NH-CH-C -NC-C0R _g is ₂ * 7 ^R 2

/ X / / VI / / I /

X stands for chloro, iodine, alkylsulfonyloxy or arylsulfonyloxy.

Amino acid of formula IV is alkylated with a-halokiselina / ester or amide / or (/ _ _C y _N u _0NIL0KSI acid / ester or amide of formula XI under basic conditions to obtain a compound of formula X. The latter is condensed by known methods, as indicated above in Method I, with the amino acid (ester or amide) of formula ΪΙ to give a compound of formula I.

The reductive cleavage of the benzyl group from the corresponding ester of formula I, in which R _g is benzyloxy and R is alkoxy, yields compounds of formula I in which P is alkoxy and R _g is hydroxy and in which R _g is alkoxy and R is benzyloxy, resulting in compounds of formula I, in which R is hydroxy and R _g is alkoxy.

Method 2 1 1 Ϊ D5 J1 fs Ϊ

RCC-NH, * XCH-CNC-COR _e ---- 4RCC-NH-CH-C I ² I ⁶ 11 'o r ₂ r ₇ o r ₂

ΪΙΙ

XII

X stands for chlorine, bromine, iodine, sulfonyloxy or aryl sulfonyloxy.

The amino acid or its derivative of the formula is

- 13 is alkylated with a suitably substituted N-haloacetyl or 7-sulfonyloxy acetyl amino acid of formula XII (preferably Rg, which is different from hydroxy) under alkaline conditions in water or in another solvent to obtain compounds of formula I.

In one case, the synthesis can be carried out in steps by condensing the amino acid ester of formula ΪΙΙ with substituted

* 3 1 ^J I ^{1 R} 3 1 ³ RCO - with - nh ₂ + X-CH-COOH —A RCO -c - i NH-CH-COOH r ₂ r ₂ III XIII X * 4 ^R 5 * 1 R ₃ 0 r ₅ 0 1 I 1 ί ii X + HN- • CH-COR ₆ _—} RC-C- II I NH- -c-c - N-C-C-fb- or ₂ * 7 ΪΙ I

N - chalosacetic acid or N -sulfonyloxy acetic acid of formula XIII to obtain the intermediate of formula X. By known methods, described is Method I, a compound of formula X may be reacted with an amino acid of formula ΪΙ or a derivative thereof to obtain a compound of the formula I. In the methods described above, an aralkyloxy moiety may be used for one or both of the substituents R and Rg to form an ester or diester.

As described above, the protecting groups can be removed by known methods.

i

The starting materials required for carrying out the processes described above are compounds known in the literature or may be prepared by known methods from known starting materials.

In the compounds of general formula I, the carbon atoms to which R p R 2 and R 2 are attached may be asymmetric. Therefore, the compounds exist in diastereoisomeric forms or as mixtures thereof. The syntheses described above may use racemates, enantiomers or diastereomers as starting materials. When diastereoisomeric compounds are obtained as a result of the syntheses performed, the latter can be separated by conventional chromatographic methods or by known fractional crystallization methods.

Although the amino acid structures, for example

Ri a ' ¹

R - C - C - NH-

Formula I-I is generally preferred in the L - and b -configurations, diastereoisomers containing D-amino acids have activity that depends on their structures and have an advantage in terms of metabolic resistance ϊπ Ϊ1Υ0 and can therefore be used in mixtures or as pure diastereoisomeric compounds.

The compounds of the invention form salts with various inorganic and organic bases, which are also within the scope of the invention. Such salts include ammonium salts, alkali metal salts, such as sodium and potassium salts, alkaline earth metal salts, such as calcium or magnesium salts with organic bases, for example dicyclohexylamine salts, N-methyl-]) -glucamine, salts with aminoninines , lysine and more. similar. Salts with inorganic or organic acids, for example hydrochloric, hydrobromic, sulfuric, phosphoric, methanesulfonic, toluenesulfonic, maleic, fumaric, camphorsulfonic, may also be prepared. Non-toxic pharmaceutically acceptable salts are preferred, although other salts may also be useful, for example in the isolation or purification of products.

The salts may be prepared by known methods by reacting the free base of the product with one or two of a suitable base or acid in a solvent or medium in which the resulting salt is insoluble or in a solvent such as water, which is then removed by distillation in vacuum or by freezing or by cation exchange of an existing salt with a cation of a suitable cation exchange resin.

The compounds of the invention inhibit the angiotensin vomiting enzyme and thus block the conversion of the decapentide angiotensin of formula I into angiotensin of formula II, which is a highly stimulating substance / blood pressure enhancer / so lowering blood pressure may result from biohazard. -especially in animals and humans whose high blood pressure is associated with the formation of angiotensin of formula II. Moreover, the converting enzyme breaks down the vasodressing agent brcykinin. Therefore, angiotensin converting enzyme inhibitors may also lower blood pressure by potentiating bradykinin. Although the relative importance of these and other possible mechanisms is yet to be established, angiotensin converting enzyme inhibitors are effective antihypertensive agents in many animal models and are also useful in clinical settings, for example in many patients with renal, malignant and underlying / essential / high blood pressure / 4 /.

Conversion enzyme inhibitors are evaluated by in vitro analysis of enzyme inhibition. For example, the method of Y. Piquilloud, A. Reinharz and M. Roth (5), in which the hydrolysis of carbobenzyloxyphenylalanylhistidylleucine is measured. It can be done in vivo, for example, in normal-pressure rats immunized with angiotensin I according to the method of J. R. Weeks and J. A. Jones (6) or in rats with high blood pressure of renal origin, such as those of o. Koletsky / 7 /.

Thus, the compounds of the invention are useful in the treatment of hypertension. They are also used in the treatment of acute or chronic congestive heart failure, in the treatment of secondary hyperaldosteronism, primary and secondary elevated pulmonary blood pressure, in renal failure, in elevated blood pressure in the renal vessels and in the treatment of cardiovascular disease and migraine. Reynold. The use of the compounds of the invention for said and other similar diseases is apparent to those skilled in the art.

For the treatment of hypertension and the clinical conditions mentioned above, the compounds of the invention may be administered in the form of tablets, capsules or elixirs for oral use, rectal suppositories.

- 17 administration, sterile solutions or suspensions for parenteral or iUramuscular administration, etc. The compounds of the invention may be administered to patients (animals or humans), in need of such treatment, at doses that provide maximum pharmaceutical efficacy. Although the dosages vary depending on the nature and severity of the disease, the weight of the patient, the special diet, the accompanying diseases, and more. Factors that the specialist in the field will have to consider, the dose is usually from 1 to 1000 mg per patient per day and may be given once or in several doses. Preferably the dose is from about 2.5 to about 250 mg per patient / day, and even more preferably the dose is from about 2.5 to about 75 mg per patient per day. The compounds of the invention may also be administered in combination with other antihypertensive agents and / or diuretics. For example, the compounds of the invention may be administered in combination with compounds such as amiloride, atenolol, bendroflumethiazide, chlorothalidone, chlorothiazide, clonidine, cryptenamine acetates, and cryptenamine tannates, deserpidine, diazoxide, guanetidyl sulphydrohydrate, hydrochloride, methylcothiazide, methyldopa, methyldonate hydrochloride, minoxidil, pergilin hydrochloride, prazosin, propranolol, rauvolfia serpentina, rescinin, reserpine, sodium nitroprusside, spironolactone, timolol, t ihlormetiazid, trimetofan, camsylate, benzthiazide, Quinet azone, triamterene, acetazolamide, aminophylline, cyclothiazide, ethacrynic acid, furozomid, meretoksilin procaine, sodium etakri sulfonate and others. as well as mixtures or combinations thereof.

Typically, the individual daily doses for these combinations may be about one-fifth of the minimum recommended clinical dose to the maximum recommended dose in cases where

- 18 give separately.

To clarify these combinations, one of the antihypertensive agents of the invention, clinically effective at doses of 2.5 - 250 mg / day, can be effectively combined in an amount of 0.5-250 mg / day with the following compounds at the indicated daily doses - hydrochlorothiazide / 15- 200 mg /, chlorothiazide / 125-2000 mg /, ethocric acid / 15-200 mg /, amiloride / 5-20 mg /, furosemide / 5-80 mg /, propanol / 20-480 mg /, timolol / 5-60 mg / and methyldopa (65-2000 mg). Triple drug combinations of hydrochlorothiazide (15-200 mg) and amiloride (5-20 mg) and the conversion enzyme inhibitor of the invention (3-200 mg) or hydrochlorothiazide (15-200 mg) and timolol (5-60 mg) and inhibitor The conversion enzyme of the invention (0.5250 mg) are effective combinations for the control of blood pressure in patients with high blood pressure. These dose limits may be tailored to the individual basis for receiving divided into daily doses and, as noted above, the dose depends on the nature and severity of the disease, the weight of the patient, the special diets and other factors.

Typically, these combinations can be formulated in pharmaceutical forms as described below.

About 1 to 100 mg of a compound or mixture of a compound of formula I or pharmaceutically acceptable salts thereof are mixed with a physiologically acceptable carrier, excipient, binder, preservative, stabilizer, flavoring agent, etc. in unit dosage form, as it is called by accepted pharmaceutical practice. The amount of active substance in these compositions or preparations is such as to provide an appropriate dosage within the above dosage range.

Examples of excipients that may be included in tablets, capsules, and the like. are the following substances: binder such as tragacanth, maize starch, gelatin; as an excipient, such as microcrystalline cellulose; as a dispersing agent, such as corn starch, pre-gelatinized starch, alginic acid, etc .; as a lubricant-magnesium stearate, as a sweetening agent such as sucrose, lactose or saccharin, as flavoring agents - peppermint, wintergreen oil or cherries. When the dose is formulated in a capsule, it may contain, in addition to the materials listed, a liquid carrier such as animal fat. Many other materials may also be present as coatings or otherwise incorporated in the modified final dosage unit form. For example, the tablets may be coated with shellac and / or sugar. A syrup or elixir may contain the active compound, sucrose as a sweetening agent, methyl and propyl parabens, such as preservatives, a dye and to impart a cherry or orange flavor.

Sterile injectable compositions may be prepared by methods known in the pharmaceutical art by dissolving or suspending the active substance in a carrier, such as water for injections, natural vegetable oils such as sesame oil, coconut oil, peanut oil, cottonseed oil, etc. . or synthetic fatty carriers such as ethyl oleate or the like. Buffers, preservatives, antioxidants, and the like may be included if necessary.

- 20 The following examples illustrate the invention by representing in particular preferred embodiments. Preferred diastereoisomers are isolated by column chromatography or fractional crystallization.

Example 1. N- (1-carboxy-2-pheylethyl) -L-alanyl-L proline.

A mixture of phenylprogesterone nicotine (753 mg) and L-alanyl-L proline (171 mg) in a mixture of water and methanol at pH 6.8 was treated with sodium cyanoborohydride (173 mg) at room temperature until the reaction was complete. The product was absorbed on a strong cation exchange resin and eluted with 2% pyridine in water to give 294 mg of crude diastereomeric product; N- (1-carboxy-2-phenylethyl) -L-alanyl-L-proline. a portion of this product was purified by gel filtration (L H-20) for spectrographic analysis. The NMR spectrum showed a broad singlet at 7.2, complex absorption from 3.0 to 4.6 multiplet at 2.1 and double doublets at 1.5 ppm.

Example 2. L1- (1-carboxyethyl) -L-alanyl-L-proline.

A solution of L-alanyl-L-proline (372 mg) and pyruvic acid (881 mg) in water at a pH of 7 was treated with 377 mg of sodium borohydride at room temperature until the reaction was complete. The product was absorbed onto a strong ion exchange resin in acid form and then eluted with a 2X-pyridine solution in water. Freeze drying provided 472 mg of N- (1-carboxyethyl) -L-alanyl-L-proline. The NMR spectrum and the mass spectrogram correspond to the structure. The NMR spectrum indicated multiplets centered at 4.5; 3.7 and 22 ppm and a pair of doublets at 1.6 ppm.

- 21 Example 3. N - N - carboxy-2-cyclohexylethyl) -L-alanylL-proline.

Process 3-Cyclohexyl-2-oxopropionic acid / cyclohexylpyrazole acid // 0.98 mg and L-alanyl-L proline (0.22 g) with sodium cyanoborohydride (0.22 g) as described above. A light colorless precipitate of solid N- (1-carboxy-2-cyclohexylethyl) -L-alanyl-L proline, 0.31 was obtained. After purification by chromatography, the mass spectrum showed peaks at 340 (molecular ions), 322, 277 , 249 and 226. The NMR spectrum showed complex absorption in the range of 4.8 to 3.6 and peaks at 2.2, 1.7 and 1.2 ppm.

Example 4. L1- (1-carboxy-5-methylhexyl) -L-alanyl-L proline.

6-Methyl-2-oxohexanoic acid (0.90 g) and L-alanyl-L-proline (0.21 g) were treated with sodium cyanoborohydride (0.21 g) as described above. A white fluffy L1- (1-carboxy-5-methylhexyl) -L-alanyl-L-proline 0.24 was obtained. After purification by chromatography the mass spectrum showed a peak at 472 (disilylyl derivative). The NMR spectrum shows absorption centered at 4.5; 3.65; 2.0, 1.6, 1.3 and 0.85 ppm.

EXAMPLE 5 N- (1-Carbony-3-methylbutyl) -L-alanyl-L · proline.

Treat 4-methyl-2-oxopentanoic acid (1.29 g) and L-alanyl-L-proline (0.32 g) with sodium cyanoborohydride (0.32 g) as described above. A fluffy white solid of L1- (N, -carboxy-3-methylbutyl) -L · -alanil-L-proline (0.40 g) was obtained. A portion of it was purified by chromatography. The mass spectrum shows a peak at 429 / molecular ion per disilyl

- 22 derivative minus methyl, 444-15 /. The NMR spectrum showed resonances centered at 4.4, 3.b, 2.1, 1.6 and at 0.95 ppm.

Example b. N- (1-carbonispropyl) -L-alanyl-L · -proline.

2-Oxoic acid (1.22 g) and L-alanylL-proline (0.37 g) were treated with sodium cyanobirhydride (0.38 g) as described above. The crude L1-L-carboxypropyl-1H-alanyl-L proline was obtained (0.42 g). Part of it was purified by chromatography for spectral analysis. The mass spectrum showed pronounced peaks at 254 (M-18) and 210 (M-62). The NMR spectrum showed complex absorption from 4.5 to 3.4 multiplet centered at 2.0 and methyl resonance centered at 1.55 and 0.95 ppm.

Example 7. N- (1-carboxy-2-methylpropyl) -L-alanyl-L proline.

A mixture of 3-methyl-2-oxobutyrate, such as the sodium salt (1.46 g), and L-alanyl-L-proline (0.40 g) was treated with sodium cyanoborohydride (0.41 g) as described above. The crude N- (L-carboxy-2-methylpropyl) -L-alanyl-L-proline was obtained by elution from an ion exchange resin (0.45 g). The product melts at 131-142 ° C. The NMR spectrum indicated complex absorption in the range 4.6 to 3.3 wide multiplet centered at 2.2 and doublets at 1.65 and at 1.1 ppm.

Example 8. N- (1,3-Dicarboxypropyl) N-alanyl-L-proline.

2-Oxoglutaric acid (1.46 g) and L-alanyl-L-proline (0.37 g) were treated with sodium cyanoborohydride (0.38 g) as described above. Crude N- (1S-dinarboxypron pyl) -L-alanyl-L-proline (0.47 g / mp) is obtained. 140-160 ° C. The mass spectrum of the silylated material shows an ion at 517 m / e, equivalent to the molecular ion for the trisilyl derivative minus methyl

- 23 / 532-15 /. The NMR spectrum confirms the structure. Methyl resonances are centered at 1.4 ppm.

Example 9. N- (1,4-dicarboxybutyl) -L-alanyl-L-proline.

2-Oxoadipic acid (1.74 g) and L alanyl-L-proline (0.41 g) with sodium cyanoborohydride (0.42 g) were treated as described above. Crude N- (1,4-dicarboxybutyl) -L-alanyl-L-proline (0.35 g / mp) is obtained. 106-132 ° C. The highest peak of the mass spectrum is 312, corresponding to the molecular ion minus water. Methyl resonances in the NMR spectrum showed a pair of doublets centered at 1.55 ppm.

Example 10. L1- (L-carboxy-3-methylbutyl) -L-alanyl-L isoleucine.

A solution of L-alanyl-L-isoleucine (150 mg) and 4-methyl-2-oxopentanoic acid, such as the sodium salt (564 mg) in water at pH adjusted to 7, was treated with 140 mg of sodium cyanoborohydride at room temperature for 6 hours. a few days. The reaction was quenched with a highly acidic ion exchange resin added to a column of the same resin and eluted with a 2X aqueous aqueous pyridine solution. Freeze-drying afforded 200 mg (84.9X/) of white powdery substance, N- (Icarboxy-3-methylbutyl) -L-alanyl-L-isoleucine. The mass spectra showed peaks at 460 for the disilylated production and at 445 for the disilyl molecular ion minus methyl (460-15). The NMR spectrum showed a broad doublet centered at 0.95 ppm, complex absorption ranging from 1.2 to 1.8 ppm and a broad weak singlet at 3.7 ppm.

Example 11. N- (1-Narboxy-3-methylbutyl) -L-alanyl-L phenylalanine.

i

- 24 Solution of L-alonyl-L · -phenylalanine (150 mg) and 4-methyl-

2-Oxpentanoic acid, sodium salt (483 mg) in water at pH adjusted to 7, was treated with 120 mg of sodium cyanoborohydride at room temperature for several days. The reaction was quenched with Dowex 50 (H +) added to the belt with the same resin and eluted with 2 N aqueous pyridine solution. Freeze-drying afforded 197 mg (88.7%) of a white fluffy substance, N- (N -carboxy-3-methylbutyl) -L-alanyl-L phenylalanil. The mass spectrum shows peaks at 551 for the trisilyl derivative minus methyl (566-15), 479 for the disilyl derivative minus methyl (494-15) and 449 for the trisilyl derivative minus -COOTMS (566-117). The NMR spectrum showed broad doubles at 0.95 and 1.5 ppm, complex absorption at 2.8-3.4 ppm (weak) and singlet at 7.1 ppm. The integration is combined with the structure, giving the correct ratio of aromatic to aliphatic protons.

Example 12. N-carboxymethyl-L-alynyl-L · -proline.

In a small flask equipped with a pH electrode are combined

1.5 g of L-alanyl-L · -proline and 1.2 ml of 4M sodium hydroxide. 0.53 g of chloroacetic acid in 1.2 ml of 2M sodium hydroxide was added. The pH was adjusted to 8-9, heated to 85 ° C, and the pH of the solution was maintained at 8-9 for 15 minutes by addition of sodium hydroxide, if necessary. 0.53 g of chloroacetic acid and sodium hydroxide are added, if necessary, for 15 minutes. A third portion of 0.53 g of chloroacetic acid was added and the pH was maintained at 8-9 for 15 minutes, kept at 85 ° C for a further 15 minutes and cooled.

The reaction mixture was passed through a column of Dowex 50 (II *), washed with water and eluted with 2X aqueous pyridine solution. The combined functions, which show a positive ninhydrin reaction, are concentrated to a small volume in vacuo and freeze-dried.

The resulting material was dissolved in several milliliters of water and loaded into a Dowex 50 (Na +) column. Elute with 0.5M-citric acid, adjusting the pH to 3.3 with sodium hydroxide. The non-reactant product appears first (ninhydrin test), well separated from unreacted alanylproline. The product fraction was concentrated in vacuo to give about 300 g.

This solution was loaded onto a Dowex 50 column (H + D. Washed with water, then eluted with a 2% solution of pyridine in water. The product fraction was concentrated in vacuo to low volume and freeze dried. 417 mg of L1-carboxymethyl-L-alanyl-L-proline are obtained.

NMR spectrum (deuterated water, methanol internal standard / 1.58 ppm / 0.1 = 6) with a small peak at 1.53 (d, I = 6) (total 3H), 1.77-2.68 (wide m, 4H), 3.63 / 3 / over 3.28-3.92 (m // 4H total), 4.05-4.72 (m wide, 2H) overlapped with the water peak 4.68.

Example 13 N- (1-Carboxyethyl) -L-alanyl-L-proline Dissolve 45 g of benzyl pyruvate 4.5 g of L-alanine in a mixture of water (115 ml) and 250 ml of p-dioxane. The pH was adjusted to 5.5 with sodium hydroxide. 9.4 g of sodium cyanoborohydride was added and stirred at room temperature for 6 days. The pH was adjusted to I with concentrated hydrochloric acid.

The solution was loaded onto a Dowex 50 (H +) column prepared in 50X-dioxane-water. Wash with 50% dioxane in water then only with water. Elute with 2X pyridine solution in

- 26 water, the product fractions were collected and concentrated in vacuo to dryness. The solid residue was triturated with water, filtered and washed with water. It is dried to give 6.8 g of L1- (Ir-carbobenzoxyethyl) -L-alanine as a mixture of diastereomers. A second crop of 1.0 g solid can be harvested from the mother liquor.

Dissolve 208 mg of the above product and 217 mg of L-proline hydrochloride benzyl ester in dry dimethylformamide. Cool to 0 ° C. 0.193 ml of diphenylphosphorylazide dissolved in dimethylformamide (DMF) was added. A solution of 0.24 ml of triethylamine in DMF was then added dropwise over 10 minutes while maintaining the temperature at 0 ° C. Stirred for 3 hours at 0 ° C and overnight at room temperature.

The mixture was diluted with ethyl acetate, washed with water and 5X sodium bicarbonate solution. It was concentrated in vacuo and chromatographed on a silica gel plate for preparative thin layer chromatography, developing with ethyl acetate. A wide band was developed at b = 0.5-0.6, eluted with ethyl acetate, and the solvent distilled to give 212 mg of the mixture of diastereoisomers of N- (1-carbobenzoxyethyl) -L-alanylL-proline benzyl ester.

135 mg of the above mixture was dissolved in methanol and water. 50 mg of 10% laladium / charcoal was added and hydrogenated at 40 psi of hydrogen at room temperature. It was filtered, concentrated in vacuo and freeze-dried to give 95 mg of the mixture of diastereoisomers of L1- (1-carboxy-ethyl) -L-alanyl-L-proline. The NMR spectrum is comparable to that obtained in Example 2, and the silylating mass spectrum, in the derivative, shows the same fragments.

Example 14. N- (1-carboxyethyl) -L-alanyl-L-proline.

Dissolve 0.75 g of L- (1-carbobenzoxyethyl) -L-alanine in pyridine and add 7.5 ml of a 1 M-triethylamine solution in pyridine. Cool and add 1.09 g of L -proline as benzyl ester hydrochloride and 0.678 g of dicyclohexylcarbodiimide. Store at 0 ° C for 20 hours. It is filtered, then concentrated in vacuo. The residue was dissolved in ethyl acetate and the solution was washed with saturated potassium carbonate solution, then with brine. The organic phase was dried, concentrated in vacuo, then chromatographed the residue obtained on silica gel and ethyl acetate-hexane was used to isolate the diastereoisomeric mixture of N- (1-carbobenzoxyethyl) -L-alanyl-L-proline.

Hydrides are typically treated with 10'-palladium / charcoal in aqueous ethanol and after treatment for isolation and freeze-drying, N- (1-carboxyethyl) -L-alanyl-L proline is obtained as a white solid.

NMR spectrum in deuterated water - 1.65 ppm / d, 6H /, 1.9-

2.6 (M, 4H), 3.5-4.2 (M, 3H), 4.3-4-8 (M, 2H).

Example 15 b1- (1-carbomethoxyethyl) -L-alanyl-L-proline. A solution of 1.4 g of methyl L-alaninate hydrochloride and 3.1 g of N-bromopropionic acid in a mixture of dioxane and water was neutralized to pH 9 with sodium hydroxide. The mixture was warmed to 70 ° C, which was maintained for 30 minutes, and the pH of the medium was maintained between 8 and 9, if necessary by the addition of sodium chloride. The mixture was charged to a Dowex 50 (H +) I column with an ion exchange resin, washed with water, and eluted with 2X aqueous pyridine solution.

- 28 Combine the fractions with the product and freeze-dry. This crude product was purified by chromatography on an ion exchange resin loaded in Djwex 50 Nolon (Na +> in 0.5 M sodium citrate buffer with pH 3.3. The product fractions were collected and concentrated in vacuo to a small volume and then re-chromatographed on Dowex 50 (H +) The product fractions were freeze-dried to give pure N- (L-carbomethoxyethyl) -alanine.

The intermediate was reacted with L-proline butyl ester using diphenylphosphoryl azide as described in Example 13. Tert was removed. butyl ester group by dissolving in trifluoroacetic acid at room temperature for 3 hours, the tetrafluoroacetic acid is distilled off and the mixture is purified on a Dowex 50 (H +) column and after elution with a 2% aqueous pyridine solution as described , N- (1-narbomethoxyethyl) -L-alanyl-L proline is obtained in the form of a mixture of diastereoisomers.

Example 16. N- (1-methoxycarbonyl-3-methylthiopropyl) -Lalananyl-L-proline.

A solution of niruvoyl-L-proline (185 mg), L-methionine, methyl ester (600 mg) and sodium cyanoborohydride (200 mg) in 20 ml of methanol was neutralized with dilute methanol sodium hydroxide solution. After standing at room temperature for 3 days, the product was absorbed on a highly acidic ion exchange resin and eluted with a 2% liter aqueous pyridine solution to give 80 mg of product. The NMR spectrum shows a methoxy group at 3.95G, a thiomethyl group at 2.2 S and CH-CH- at 1.55 and 1.7 S '· The silylated mass spectrogram

W

- 29 material shows the expected molecular ion at 404 m / e.

Example 17. N-1- (1/5) -carboxy-3-methylthiopropyl (-alanyl) -1-proline.

Solution of N- [1- (6) -methoxycarbonyl] -3-methylthiopropyl (DL-alanyl-L · -proline) Ί27.5 mg; 0.384 mM / in 2 ml of water was treated with 8 nitrogen atmosphere with 7.82 ml of 0.100 I sodium chloride (0.782 mM) and stirred for two and a half hours at room temperature. The product was absorbed from the reaction mixture on 30 ml of Dowex 50 (H +) and eluted with 4% aqueous pyridine solution to give 73.5 mg of product, which was further purified on an Ij H-20 column to give 55.7 mg product. The NMR spectrum in deuterated water showed thiomethyl at 2.1; CH-CH ₃ at 1.5 and 1.6 P and lack of methyl ester. The mass spectrogram of the silylated material shows the expected molecular ion at 462 m / e.

Example 18 N- (T-methoxycarbonyl-2- (3-indolyl) -ethyl) alanyl-L-proline.

Work as described in Example 16 to condense tryptophone methyl ester with pivaloyl-L-proline in the presence of sodium cyanoborohydride to give N- (1 * -methoxycarbonyl-2- (3-indolyl) -ethyl) -alanil -1? -Proline.

The NMR spectrum in deuterated chloroform showed aromatic protons at 6.9 to 7.7; protons adjacent to the aromatic nuclei and adjacent to the nitrogen at 2.8 to 3.9; aliphatic methylene proton at 1.4 to 2.7 and alanine methyl at 1.0 to 1.4. A mass spectrogram of the silylated material shows an ion at 516 m / e in accordance with the methyl group lost dissolved material.

Example 19 N- (1/5) -carboxy-2- (3-indolyl) -ethyl] -Ob olanyl-L-proline.

- 30 Work as described in Example 17, hydrolyzing said product to give the expected diacid. The NMR spectrum in DzO-c ^ PYR · showed five aromatic protons at 6.8 to 7.7; 7 protons adjacent to the aromatic nucleus and adjacent to nitrogen at 2.8 to 7.4 and 7 aliphatic protons at 1.0 to 2.2 pounds according to the expected structure. The mass spectrogram of the silylated material shows a peak at 431 m / e, interpreted as a displaced minisilylated ion that lost a methyl group.

Example 20 N- (1-carboxy-3-phenylpropyl) -L-alanyl-L thiazolidine narboxylic acid.

Combine tert.-Boc-alanine (1.8 g) and L-thiadolidin-4 carboxylic acid in the form of benzyl ester, hydrochloride (2.6 g) in methylene chloride. At 0-5 ° C, it was treated with triethylamine (1.4 ml), then dicyclohexylcarbodiimide (DSS) (2.3 g) in methylene chloride, then allowed to stand overnight. After filtration and washing of the filter with water and sodium bicarbonate solution, the solvent was distilled off and the product was chromatographed on silica gel 6-60 (E. Merc) in ethyl acetatehexane. The solvent of the collected fractions containing the product was distilled in vacuo. The benzyl ester is hydrolyzed in acetonitrile-water at pH 13.5 (sodium hydroxide) for one hour at room temperature. The medium was neutralized to pH 8 with hydrochloric acid, washed with ether, concentrated the aqueous layer in vacuo and then freeze-dried. It is removed from the tert-butoxycarbonyl protecting group in a 4 M solution of hydrogen chloride in ethyl acetate. The product was triturated with ether, filtered and dried to give L-alanyl-L-thiazolidine-4-carboxylic acid. 0.385 g of this product was condensed with 1.88 g of 2-oxo-4-phenylbutyric acid

- 31 acid in water using 0.354 grams of sodium cyanoborohydride according to the method described in Example 24 to give 0.53 g of a mixture of the diastereoisomer of N-carboxy-3-phenylpropyl-N-alanyl-L -4-thiazolidine carboxyl koselina. The NMR spectrum in deuterated water and deuterated sodium hydroxide contains a doublet at 1.2 ppm (3H), singlet at 7.1 / 5 N /, absorption width in the range from 1.6 to 2.0 / 2H /, wide absorption in the range from 2.2 to 4.1 and wide water peak at 4.6 ppm. The mass spectrum of the silylated product shows the molecular ion of the disilylated derivative at m / e = 55b.

Example 21 N- (L-Carboxy-3-phenylpropyl) -L-alanyl-L pipecolic acid.

By replacing the thiazolidine carboxyl ester of Example 20 with L-pipecolic acid, methyl ester, hydrochloride (1.8 g), the title compound can be obtained using the method described in Example 20.

The NMR spectrum in deuterated methanol showed a wide multiplet at 1.3 to 1.9 ppm (9H8), a singlet at 7.22 (5H), and a series of multiplets in the range 2.0-4.8 ppm. The spectrum of the silylated material shows a peak at m / e = 580 for the disilylated molecular ion.

Example 22 N- (N-carboxy-3-finylpropyl) -L-alanyl-L N-methyl-alanine.

Replacing the thiazolidine carboxyl ester of Example 20 with b-N-methylalanine, methyl ester, hydrochloride (1.5 g) by the method described in the same example, the title compound was obtained.

Example 23. N- (1-Narboxy-1-methylethyl) -L-alanyl-L proline.

- 32 The title compound may be obtained by mixing 7.7 g of 2-bromo butyric acid, benzyl ester, 2.4 g of L-alanyl-L-proline, tert. butyl ester, 7.0 g of silver oxide in 40 ml of benzene. After refluxing for 24 hours, an additional 7.7 g of the bromester and 7.0 g of silver oxide were added and reflux continued for another 24 hours. Cooling, filtration, distillation of the solvent and the product diester were isolated by conventional chromatographic methods. After cleavage of tert. butyl ester group in trifluoroacetic acid and benzyl group by catalytic hydrogenation by known methods yields the desired free acid.

Example 24 N- (1-carboxy-3-phenylpropyl) -L-alanyl-L proline.

A mixture of 4-phenyl-2-oxobutyric acid (1.49 g) and L alanyl-L -proline (0.31 g) in water at pH adjusted to 7.5 with sodium hydroxide was treated with sodium cyanoborohydride (0.32 g) for one night. The product was absorbed on a highly acidic ion exchange resin and eluted with a 2 N-solution of pyridine in water to give 0.36 g of crude diastereomeric product, N- (N-carboxy-3-phenylpropyl) -N-alanyl-Lproline. Part of it was purified by gel filtration (L H-20) for the purpose of spectrographic analysis. The NMR spectrum in dimethyl sulfoxide shows aromatic hydrogen at 7.20, a wide singlet at 4.30 wide multiplet in the range 3.0 to 3.9, 2.67 and 1.94 and a doublet at 1.23 and at 1.15. The mass spectrum shows a molecular ion at 492 m / e for dimethylsilylated materials.

Example 25. N- (1-carboxy-3-phenylaropyl) -L-alanyl-L proline.

- 33 Grind and sift XAD-2 polyester resin (Rohm and

Haas Co.), the fraction was separated with 200-400 mesh and 440 ml were loaded into a chromatographic column. Equilibrate with 0.1 M ammonium hydroxide and a mixture of water-methanol 95 to 5 (v / v). 350 mg of N- (1-carboxy-3-phenylpropyl) -L-alanyl-L-proline prepared as described in Example 24 dissolved in 10 ml of the same solvent are loaded into the column. The first isomer leaves the column within a volume range of 375-400 ml of the eluate. The second isomer is in the range of 440-480 ml with an intermediate fraction containing a mixture of isomers. Freeze drying of the fraction containing the first isomer gave 130 mg of a white solid. Recrystallization from 1 ml of water at pH adjusted to 3 gave 94 mg of white needle crystals, m.p. Mp 148-151 ° C. It is the most active isomer and has a $, $, 3 configuration, as determined by X-ray analysis ΛΖ ( _α = -67.0 ° /0.1 M HCl) after vacuum drying over diphosphorus pentoxide. The NMR spectrum in dimethylsulfoxide shows a single doublet for methyl protons at 1.22 ppm. Freeze drying of the fraction containing the second isomer yielded 122 mg of a white solid. Recrystallization of 103 mg of it from 2.5 ml of water with pH adjusted to 3 results in 64 mg of light white crystals, m.p. 140-145 ° C d., / C // d = -101.6 ° /0.1 M HCl / after drying. The NMR spectrum in dimethyl sulfoxide shows the methyl doublet at 1.17 ppm.

Example 26. N 1- (1/5) -ethoxycarbonyl-3-phenylpropyl] -L alanyl-L-proline.

Ethyl 2-oxy-4-phenylbutyrate (1.0.03 g) and Lalananyl-L-proline (0.19 g) were dissolved in a mixture of 1 to 1 ethanol / water. To the resulting solution for 2 hours at room temperature

- 34 A solution of sodium cyanoborohydride (0.19 g) in ethanol was added dropwise. When the reaction is complete, the product is absorbed onto a highly acidic ion exchange resin and eluted with a 2% aqueous pyridine solution. The product-rich Francia was freeze-dried to give 0.25 g of crude N-ZJ-ethoxycarbonyl-3-phenylpropyl] -L-alanyl-L-proline. The mass spectrum shows a molecular ion at 448 m / e for the monosilylated material. Chromatography provides the desired isomer.

Example 27. N1- (1 -aminocarbonyl-3-phenylpropyl) -L-alanylL-proline.

This was done as described in Example 26, which may condense 2-oxo-4-phenylbutyramide and L-alanyl-L-proline in the presence of sodium cyanoborohydride to give N- (1-aminocarbonyl-3-phenyl-propyl) N-alanyl-L-proline.

Example 28. N- (1-carboxy-3-phenylpropyl) -L-alynyl-L tryptophan.

Work as described in Example 24, which can condense 2-oxo-4-phenylbutyric acid with L-alanyl-L tryptophan in the presence of sodium cyanoborohydride to give N- (1-carboxy-3-phenylpropyl) - L-alanyl-L-tryptophan.

Example 29. N- (1-carboxy-3-phenylpropyl) -L-alanyl-L -4hydroxy-proline.

In the procedure described in Example 24, 2-oxo-4-phenylbutyric acid and L-alanyl-4-hydroxyproline were condensed in the presence of cyanoborohydride to give 1- (1-carboxy) -

3-phenyl-prolyl-N-alanyl-L -4-hydroxylroline.

The NMR spectrum in deuterated methanol shows a doublet centered at 1.53 ppm (3H), singlet at 7.13 (5H), a series of multiplets in the range 2.0 to 4.7 ppm. The mass spectrum of the silylated product shows the molecular ion of the trisilyliron product at m / e = 580.

Example 30. N- / Ι-κορ6οκ with and-3-phenylpropyl / L · -serinyl-L proline.

In the method described in Example 24, 2-oxo-4-phenylbutyric acid and L-serinyl-L-proline were condensed in the presence of sodium cyanoborochlor to give L1- (1-carboxy-3-phenylpropyl) -L-serinyl-L - proline.

The mass spectrum shows a molecular ion at 580 m / e for trisilylated species. The NMR spectrum in deuterated water confirms the structure.

Example 31 N- (N-carboxy-3-phenylpropyl) -L-phenylalanylL-proline.

In the method described in Example 24, 2-oxo-4-phenylbutyric acid and L-phenylalanil-L-proline were condensed in the presence of sodium cyanoborohydride to give N- (1-carboxy-3-phenylpropyl-L-proline).

The mass spectrum shows one ion at 406 m / e for the molecular ion minus water (424-18). The NMR spectrum in deuterated water corresponds to the structure.

Example 32 b1- (1-carboxy-3-phenylpropyl) -L-cysteinyl-L proline.

In the method described in Example 24, 2-oxo-4-phenylbutyric acid can be condensed with L -5-benzylcysteinyl-L proline in the presence of sodium cyanoborohydride. The product can then be treated with sodium in liquid ammonia to give L1- (1-carboxy-3-phenylpropyl) -L-cysteinylL-proline.

Example 33 N- (1-carboxy-3-phenylpropyl) -L-histidinyl-36 L-leucine.

By the method described in Example 24, they can be condensed

2-Oxo-4-phenylbutyric acid with L-histidinyl-L · -leucine in the presence of sodium cyanoborohydride to give

N- (1-carboxy-3-phenylpropyl) -L-histidinyl-L-leucine.

Example 34 N- (1-carboxy-3-phenylpropyl) -L-phenylalanil-L arginine.

By the method described in Example 24, they can be condensed

2-Oxo-4-phenylbutyric acid with L-phenylalanyl-L-arginine in the presence of sodium cyanoborohydride to give N- (I-carboxy-3-phenylpropyl) -L-phenylalanyl-L-arginine.

Example 35 N- (1-carboxy-3-phenylpropyl) -L-phenylalanylL-tryptophan.

By the method described in Example 24, they can be condensed

2-Oxo-4-phenylbutyric acid with L · -phenylalanyl-L-tryptophan in the presence of sodium trian cyanoborohydride to give N- (1-carboxy-3-phenylpropyl) -L-phenylalanyl-L-tryptophan.

Example 36 N- (1-carboxy-3- (3-indolyl) propyl) - L alanyl-L-proline.

In the method described in Example 24, 4- (3-indolyl) -2-oxo-butyric acid and L-alanyl-L-proline were condensed in the presence of sodium cyanoborohydride to give N- (1-carboxy-3- / 3-). indolyl (propyl) N-alanyl-L-proline. The mass spectrum of the trisilylated material shows a molecular ion at 603 m / e and an M ♦ -15 peak at 588 m / e.

Example 37 b1- (1-carboxy-3-phenylpropyl) -N-alanyl-3,4-dihydroproline. Mixture of 3,4-dihydroproline (2.3 g), tert. Boc-b alanine. The N-hydroxysuccinimide ester (7.2 g) and sodium carbonate (2.5 g) are stirred in a mixture of dioxane-water at

- 37 0 ° C overnight. The pH of the medium was neutralized to 8 with hydrochloric acid. Concentrate to a small volume in vacuo and freeze-dry. The tert. Boc protecting group with trifluoroacetic acid in the usual manner and the product was chromatographed on elution with 2H aqueous pyridine using Dowex 50 (H +) by the method described in Example 2. The dipeptide was isolated by freeze-drying. The resulting product was reacted with 2-keto-4-phenylbutyric acid according to the method described in Example 24 to afford the product as a mixture of diastereoisomers. The spectrum shows a molecular ion at 490 m / e for the disilylated species.

The diastereomeric product (140 mg) obtained above was separated into components by chromatography on XAD -2 resin in the manner described in Example 25. The major component (70 mg), which first left the column, was Ζα (c = 1.3 methanol). The mass spectrum of each component shows a molecular ion at 490 m / e for the dimethylsilylated species.

Example 38 N- (1-Carboxy-3-phenylpropyl) -1 galanyl-2methyl-thiazolidine-4-carboxylic acid.

This compound can be prepared by the method described in Example 37, replacing (2.3 g) / 3,4-dihydroproline (2.9 g)

2-methyl-thiazolidine-4-carboxylic acid.

Example 39. L1- (1-Carboxy-3-phenylpropyl) -1'-alanyl-Lmethylananine.

This compound can be prepared by the method described in Example 37, replacing 2.3 g of 3,4-dihydroproline with 1.8 g of 2-methylalanine.

Example 40. Dry capsules containing 50 mg each

- 38 active ingredient.

Each capsule (size 1) of 200 mg contains the following components, in mg: 1 N- (1-carboxy-2-phenyl fly N) -L-alanyl-L · proline 50; lactose 149; Magnesium stearate 1.

Grind N- (N-carboxy-2-phenylethyl) -L-alanyl-L proline to a particle size of 60, and then lactose and magnesium stearate are passed through a No. 60 sieve onto the powdered active ingredient. The ingredients are mixed for 10 minutes and then filled into a dry gelatin capsule No. 1.

Example 41. N - [- / 1'-Ethoxycarbonyl-3-phenylpropyl / -L-alanylL-prol.

A solution of L-alanyl-L-proline (7.7 g) and ethyl-2-oxo-4-phenylbutyrate (42.6 g) in 140 ml ethanol was stirred with 64 g of powder molecular sieve at room temperature for half an hour. Then a solution of sodium cyanoborochloride (2.6 g) in 40 ml of ethanol was slowly added over 6 hours. After filtering the sieve, the reaction mixture was concentrated in vacuo to a small volume. The resulting residue was partitioned between chloroform and water. The pH was adjusted to 8.5 and the chloroform layer was separated and discarded, the aqueous layer was increased to pH 2.7 and the product was extracted with chloroform. The chloroform extract was dried over sodium sulfate and concentrated in vacuo to give 10.4 g of mixed diastereomers. High-performance liquid chromatography (HPLC) indicated that the major product was the desired- (1- (S) -ethoxycarbonyl-3-phenylpropyl) -L-alanyl-L-proline

The NMR spectrum showed aromatic absorption at 7.1 µL and diastereomeric methyl as a multiplet centered at 1.3cP.

The mass spectra showed a molecular ion at 376 m / e and strong

- 39 peaks at 358 m / e (M ♦ -HgO).

Example 42 N- (1- (S) -ethoxycarbonyl-3-phenylpropyl) -L alanyl-L · -proline, maleate salt.

A solution of N- (1-ethoxycarbonyl-3-phenylpropyl) -1'-alanyl-L-proline, a mixture of isomers (13.8 g) in 69 ml of acetonitrile was treated with 4.25 g of maleic acid in 69 ml of acetonitrile. After stirring for 1 hour at room temperature, the solid was filtered off, washed with acetonitrile and dried to give 8.4 g of maleate salt, m.p. 141-145 ° C. High performance liquid chromatography (HPLC) established a product purity of 96%. The crude maleate salt was recrystallized from acetonitrile to give 7.1 g of N- (S) -ethoxycarbonyl-3-phenylpropyl) -L-alanyl-L-proline, maleate, m.p. 148150 ° C, 99% purity (HPLC).

Example 43

A. N- (1-Ethoxycarbonyl-3-phenylpropyl) -L-alanyl-L proline.

Mixture of 0.814 g of L-alanyl-L-proline, 0.206 g of ethyl 2-oxo-

4-Phenylbutyrate and 1.6 g of molecular sieve in 10 ml of ethanol were hydrogenated at room temperature at 40 lbs of pressure with about 1 gram of 10H-palladium / carbon as catalyst. After stopping the flow of hydrogen, the crude product was obtained by filtration and concentration, then absorbed onto an ion exchange resin (Dowex 50, H +) and eluted with a 2% pyridine solution in water to give 0.224 g of N- / I 'ethoxycarbonyl-phenylpropyl] -L-alanyl-L-proline. HPLC shows that the ratio of isomers is 55 to 45.

B. N- (1-Ethoxycarbonyl-3-phenylpropyl) -E-alanyl-L proline, a maleic acid salt.

- 40 A mixture of 3 g of L-alanyl-L-proline, 5 g of ethyl-2-oxo-4-phenyl-butanoate, 13 g of a molecular sieve and 3.6 g of Raney nickel in 85 ml of ethanol is hydrogenated at 25 ° C and a pressure of 40 psi of hydrogen until it stops absorbing hydrogen. The solids were filtered off, washed with 80 ml of ethanol and the filtrates collected. HPLC analysis showed that the ratio of diastereoisomers in the mixture was 87 to 13 in favor of the desired product. The ethanol was removed in vacuo to give an oil which was dissolved in 60 ml of water and 20 ml of ethyl acetate. The pH of the resulting biphasic mixture was stirred to

8.6 using 50% sodium hydroxide. The layers were separated and the aqueous phase was extracted twice with 20 ml of ethyl acetate. The pH of the aqueous phase was adjusted to 4.25 with hydrochloric acid, 12 g of sodium chloride dissolved in the solution were added and the product was extracted five times with 12 ml of ethyl acetate. The extracts were collected and dried over sodium sulfate. The polonium product N1- (1R) .57-ethoxycarbonyl-3-phenylpropyl (1S) alanyl-L-proline was quenched in the form of its maleic acid salt by the addition of 1.86 g of the acid. After stirring for 4 hours, the salt was filtered off, washed with ethyl acetate, and after drying 5.2 g of pure product, m.p. 150-151 ° C.

Example 44. L1- (1-Benzyloxycarbonyl-3-phenylpropyl) -L alanyl-L · -proline.

A solution of L-alanyl-L-proline (167 g) and benzyl 2-exo-4phenylbutyrate (1.20 g) in 5 ml of ethanol was stirred at room temperature with a 3 g powder molecular sieve, type 4A. Sodium cyanoborohydride (75 mg) was then added portionwise over 3 hours. The product is purified by absorption

- 41 on a strong cation exchange resin and eluted with 2X pyridine solution in water. After passing through a gel filtration column (b-H-20), 220 mg of N-benzyloxycarbonyl-3-phenylpropyl) -L-alanyl-L-proline were obtained in the form of a mixture of isomers. Thin layer chromatography on silica gel eluting with 1 ethyl acetate, 1 n-butanol, 1 water, 1 acetic acid showed one major spot at R _F 0.71. The isomers were separated using reverse phase HPLC to afford N1- (R) -benzyloxycarbonyl-3-phenylpropyl-L-alanyl-L-proline.

Similarly, L1-acetylaminoethyl-2-oxo-4-phenylbutyrate and L-alanyl-L-proline are reduced with sodium cyanoborohydride to give N- (1- (2-acetylamino) ethoxycarbonyl-3-phenylpropyl) -L-alanyl. -L -proline.

Similarly dimethylaminoethyl 2-oxo-4-phenylbutyrate and L-alanyl-L-proline give N- (1- (2-dimethylamino) -ethoxycarbonyl-3-phenylpropyl) -L-alanyl-L-proline.

Similarly, benzyl 2-oxo-5-methylhexanoate and L-alanyl-L-proline yield N- (1-benzyloxycarbonyl-4-methylpentyl) -L-alanyl-L-proline.

The mass spectrum of the silylated material shows peaks at 510 (M ⁺ very weak), 495 (M ⁺ -15), 464, 420, 419, 385 and 296 m (e) (base peak).

The NMR spectrum is consistent with the structure.

EXAMPLE 45 B1 - [.butoxycarbonyl-3-phenylpropyl] -L-alanylL-proline.

A solution of GT-benzyloxycarbonyl-L-alanyl-L-proline butyl ester (452 mg) in 5 ml of benzene was hydrogenated on 150 mg палаθ /-ene palladium / charcoal until the nitrogen protecting group was removed. After filtration and removal of the solvent L-alanyl-L proline 3 ° the butyl ester was dissolved in 8 ml of tetrahydrofuran

42 and treated with 1.41 g of butyl 2-oxo-4-phenylbutyrate and 3 g of powder molecular sieve. Sodium cyanoborochloride (150 mg) was added portionwise over several hours, after which the mixture was stirred at room temperature overnight. After filtration and concentration in vacuo, the resulting residue was treated with 25 ml of trifluoroacetic acid at room temperature for 2 hours. After removal of the acid, the product was purified by absorption on an ion exchange resin and by gel filtration (b H-20). Concentration and drying of the product containing fractions afforded 182 mg of b1- (1-butoxycarbonyl-3-phenylpropyl) -L-alanyl-L-proline as a mixture of isomers. Thin layer chromatography (silica gel, 1 ethyl acetate, 1 butanol, 1 water, 1 acetic acid) showed two spots, R _F 0.67 and 0.72. The mass spectrum shows peaks at 548 (disilylated molecular ion) and at 476 (monosilylated molecular ion). The isomers were separated using reverse-phase high-performance liquid chromatography to obtain N- (S) -butoxycarbonyl-3- (phenylpropyl) N-alanyl-L-proline.

Example 46 1- [1-Ethoxycarbonyl-3-phenylpropyl] -L-alanylL-proline, ethyl ester.

A solution of 0.63 g of N- (R) -carboxy-3-phenylpropyl) -N-alanyl-L-proline in 9.7 ml of ethanol was saturated with hydrogen chloride gas at 0 ° C. After standing at room temperature overnight, the hydrochloric acid and ethanol were removed in vacuo to give a pale yellow oil which was purified by gel filtration (L H-20 column). The yield of N- (1-ethoxycarbonyl-3-finylpropyl-E-alanyl-L-proline, ethyl ester is 0.39 g; thin-layer chromatography shows one spot. The NMR spectrum shows two ethyl groups to the aromatic ring. The mass spectrum shows a molecular ion at 404 m / e.

Example 47. L1- (1-carboxy-3-phenylpropyl) -L-alanyltrans-4-methoxy-L -proline.

Methyl trans-4-methoxy-L-proline hydrochloride from L-hydroxyproline was prepared by the method of E.Ac | am $ eb αΐ. 1.Viol Seven. 208,573 (1954) by esterifying with methanolic hydrochloric acid in a known manner. The following is reacted with Boc-L-alanine in methylene chloride with dicyclohexylcarbodiimide as described above, purification of the intermediate Boc-L-alanyl-L-methoxy Pzo-OMe by chromatography on silica gel eluting with a mixture of ethyl acetate and hexane I / Ι . Hydrolysis of the ester with sodium hydroxide in acetonitrile-water, adjusting the pH to 7.5, freeze-drying, removing the protecting group of the amine in 4M hydrochloric acid in ethyl acetate in a known manner. Condensation of 0.54 g of this L-alanyl-L-TpaHC-4-Methoxy-L-proline with 2.0 g of 2-oxo-4-phenylbutyric acid in 6 mm of water using 0.43 g of sodium cyanoborohydride by the method described in Example 24 Isolation of the product described in the same example to give 0.92 g of a mixture of diastereoisomers of N- (Lcarboxy-3-phenylpropyl) -L-alanyl-trans-4-L-proline.

The NMR spectrum in deuterated water shows a doublet centered at 1.58 ppm (3H), singlets at 3.37 (3H) and 7.35 ppm (5H), complex absorption in the 1.9-3.5 range and wide multiplet at 4.0-4.6 ppm. The mass spectrum shows pronounced peaks at m / e = 360 (M-18) and at 256 (M-122).

Example 48 N- (1-benzyloxycarbonyl-3-phenylpropyl) -L alanyl-1rans-4-methoxy-L-proline.

By joining L-alanyl-L-4-methoxy-syproline,

- 44 obtained in Example 47, with benzyl-2-oxo-4-phenylbutyrate in ethanol using sodium cyanoborohydride according to the method described in Example 44, can be obtained from the diastereoisomers of b1- / 1'-benzylsulfonyl-3- phenylpropyl-L-alanyl-trans-4-methoxy-L-proline. The isomers can be separated using reverse phase HPLC to give N- (I) (S) benzyloxycarbonyl-3-phenylpropyl) -L-alanyl-trans-4-methoxy-L-proline.

Example 49. N1- (1-Benzylaminocarbonyl-3-phenylpropyl) -L alanyl-L-proine.

2-Oxo-4-phenyl-butyric acid benzylamide was obtained by dissolving 3.0 g of acid, 2.4 ml of benzylamine and 4.7 ml of diphenylphosphorylazide in 60 ml of cold dimethylformamide and adding dropwise 2.6 ml of triethylamine in dimethylformamide while maintaining the temperature. about - 10 ° C for two and a half hours. Standing at room temperature overnight, dimethyl formamide was distilled in vacuo and the residue partitioned between water and ethyl acetate. The contents of the organic layer were chromatographed on silica gel, eluting with ethyl acetate / hexane at a ratio of 1 to 4. From the fractions containing the product, the solvent was evaporated to give 2.2 g of crystalline n-benzyl-2-oxo-4-phenylbutyromide. 1.26 g of this product was reacted with 0.19 g of L-alanyl-L-proline using 0.125 g of sodium cyanoborohydride in ethanol by the method described in Example 44. The crude product was purified by gel filtration (L H-20) to give a mixture of the diastereoisomers of N- (1-benzyl-aminocarbonyl) -3-phenylpropyl) -L alanyl-L-proline. The NMR spectrum in deuterated chloroform shows a doublet at 1.1 ppm (3H), a close pair of singlets at 7.3 (UN).

- 45 and complex absorption in the range of 1.6-2.3 (6H), 2.3-2.9 (2H), 2.9-3.8 (4H) and 4.0-4.6 (3H).

The mass spectrum of the silylated material shows pronounced peaks at m / e ~ 509 (monosilylated derivative) and at 581 (dissolved derivative).

Example 50 N- (R) -carboxyl-3-phenylpropyl N-L-alanyl-L N-methylphenylalanine.

By replacing the thiazolidine carboxyl ester by example

20, with N-methyl-L-phenylalanine, methyl ester, is obtained

L-alanine-N-methyl-L-phenylalanine. By condensing 0.85 g of this product with 0.32 g of 2-oxo-4-phenylbutyric acid using 0.64 g of sodium cyanoborohydride in water as described above, acidifying the mixture to pH 1.5 and extracting with ester, after t8a distillation ether, dissolving the residue in 70% methanol in water and chromatographing on Dowex 50 (H +) ^{8 the} same solvent and eluting with 3X pyridine solution in the same solvent mixture, collecting product containing fractions, concentrating, freeze drying. Purification on LH-20 in methanol yielded 0.54 g of a mixture of N- (1-carboxy-3-phenylpropyl) -L-alanyl-L-N-methylphenylalanine isomers. The NMR spectrum (deuterated water, deuterated sodium hydroxide) shows a doublet centered at 1.18 ppm (3H), two overlapping singlets at 7.3 (10H), singlet at 2.95 (3H), and wide multiplet absorption in the intervals 1.4-2.1 and 2.3. -4.4. The mass spectrum of the silylated material shows the molar ion of the disilylated material at m / e = 556.

Example 51. L1- (1-Ethoxycarbonyl-3-phenylpropyl) -L-alanyl-L proline amide.

The L-alanyl-L-proline amide is obtained by coupling

- 46 but tert-Boc-L-alanine with L-proline amide by known methods using dicyclohexylcarbodiimide in a mixture of 4 to 1 methylene and chloroform to dimethylformamide. Purification of the intermediate tert. Boc-L-alanyl-L-proline amide by chromatography on L H-20 in methanol, then removing the protective tert. Boc group in 4 M hydrochloric acid solution in ethyl acetate. Binding 0.5 g of then L-alanyl-L-proline amide hydrochloride in 10 ml of absolute ethanol, neutralized with one equivalent of triethylamine, with 2.4 g of ethyl 2-oxo-4-phenylbutyrate using a molecular sieve and 0.30 g of sodium cyanoborohydride the method described in Example 41. In this example, the product was taken up in the chloroform extract at pH 8.5, concentrated in vacuo, the residue dissolved in 50X ethanol-water mixture, chromatographed on Dowex 50 (H +) filled with 50X ethanol mixture and eluted with a 2% pyridine solution in the same solvent. The fractions containing the product were collected and further purified by chromatography on LH-20 in methanol. The solvent was distilled in vacuo to give 0.40 g of N- (1-ethoxycarbonyl-3-phenylpropyl) -L-alanyl-L-proline amide as a mixture of diastereomers. The NMR spectrum in deuterated chloroform shows a triplet overlapping doublet at 1.1-1.5 ppm (6H), a series of five multiplets in the range of 1.5-4.7 ppm (15 N) and a singlet at 7.17 ppm (5 N). The mass spectrum of the silylated product shows marked peaks at m / e = 477 (monosilylated production) and at 519 (disillusioned production).

Example 52. N- [1'-Carboxy-3-phenylpropyl] -L-alanyl-L proline amide.

The L-alanyl-L-proline amide obtained in Example 51 was reacted with 2-oxo-4-phenylbutyric acid using sodium cyanoborohydride in a 50% mixture of ethanol and water according to the method described in Example 2. After elution from ion exchange resin, concentration in vacuo to low volume, pouring with water and freeze-drying afforded N- (1-carboxy-3-phenylpropyl) -L-alanyl-L-proline amide in the form of a mixture of diastereoisomers.

The NMR spectrum showed aromatic absorption at 7.35 ί and a pair of doublets for diastereomeric methyl at 1.55 (P.

Example 53 N- (N-) - (6) hydroxyaminocarbonyl- (3-phenylpropyl) -L-alanyl-L-proline.

To a cold solution of 0.19 g of L- (1-ethoxycarbonyl-3-phenylpropyl) -L-alanyl-L-proline, the maleate salt obtained in Example 42, in 1 ml of ethanol was added 0.85 g of potassium hydroxide in 0.57 ml of ethanol. A suspension of 0.07 g of hydroxylamine hydrochloride in 0.9 ml of ethanol containing 0.060 g of potassium hydroxide is then added dropwise. The mixture was maintained in an ice bath for two hours, then at room temperature overnight. The supernatant was decanted, diluted with 10 ml of water, the pH was adjusted to 2.5 with hydrochloric acid and washed with chloroform. After neutralization, freeze-drying and purification by chromatography on an XAD -2 resin with a gradient of 0.1 M ammonium hydroxydethanol, L- (1- (1-X) - hydroxyaminocarbonyl-3-phenylpropyl) -L-alanyl-proline is obtained. The mass spectrum of the silylated product shows an ion at m / e = 579 for trisilylation production and the NMR spectrum is consistent with the structure.

Example 54 N- (1-carboxy-3-methylbutyl) -1H-alanyl-L · tryptophan.

Sodium salt solution on 4-methyl-3-oxo-pentanoene

48 acid (414 mg) and L-alanyl-L-tryptophan (150 mg) in water at pH adjusted to 7 with sodium hydroxide were treated with sodium cyanoborohydride (103 mg) at room temperature for several days. The product was absorbed onto a highly acidic ion exchange resin and eluted with a 2 ^ pyridine solution in water. Product-rich fractions were collected and freeze-dried to give 189 mg of a light white solid. The mass spectrum shows a molecular ion at 389 m / e and peaks at 187 m / e and at 158 m / e for the following fragments:

dream _? sn

I 1

H00C-CH-IIH-CH

The NMR spectrum in deuterated water confirms the structure.

Example 55. N- (1-carboxy-3-methylbutyl) -L-histidylL-leucine.

In the method described in Example 54, 4-methyl-2-oxopyanoic acid and L-histidyl-L-leucine are condensed in the presence of sodium cyanoborohydride to give Ν- / Ι-καρ6οκοΗ-

3-methylbutyl-L-histidyl-L-leucine. In this case, the product is eluted from the ion exchange resin with 10X ammonia solution. The mass spectrum shows a molecular ion at 408 m / e for the disilylated species minus 18. The NMR spectrum confirms the structure.

Example 56. N- [1-Carboxy-3-methylbutyl-L-phenylalanyl] -arginine.

In the method described in Example 54, 4-methyl-2-oxopentanoic acid and L-phenylalanil-L-arginine are condensed

49 in the presence of sodium cyanoborohydride to give N- (1 N -carbony-3-methylbutide) -L-phenylalanil-L-arginine. The product was eluted from the ion exchange resin with a 10% ammonia solution. The NMR spectrum of deuterated water showed singlet at 7.2 (5H), broad trillet, concentrated at 3.0 (6H), wide multiplet at 1.3 (6H) and broad doublet at 0.9 (6H).

Example 57. L1- (1-carboxy-3-phenylpropyl) -L-lysyl-L proline.

According to the method described in Example 54, 2-oxo-4-phenylbutyric acid and tert. Boc-L-lysyl-L-proline is condensed in the presence of sodium cyanoborohydride. Essentially all tert. Boc-protecting groups are cleaved but the product is absorbed on the highly acidic ion exchange resin. The crude L1- (1-carboxy-3-phenyllropyl) -L-lysyl-L-proline was eluted from the resin with a 10% solution of ammonia, freeze dried and purified by gel filtration (LH-20). The minor peak for tert.-Boc protons in the NMR spectrum disappears when the product is treated with ethyl acetate with 4 N hydrochloric gas. The NMR spectrum of the resulting hydrochloride salt of the product is consistent with the structure. The mass spectrum shows a molecular ion at 693 m / e for trisilylated species. Chromatography on the PAP-2 resin using 3.5 H acetonitrile in 0.1 M-ammonium hydroxide afforded N-X- (1 / S) -carboxy-3-phenylpropyl) -L-lysyl-L-proline.

B. b1 - N- [1- (1/5) -naryloxy-3-phenylpropyl] -L-lysyl-L-proline.

In the method described in Example 54, 2-oxo-4-phenylacetic acid and N-tert-Boc-L-lysyl-L-proline were condensed in the presence of sodium cyanoborohydride. The product was absorbed on a highly acidic ion exchange resin and eluted with 2X-ene

- 50 solution of pyridine in water. The product-rich fractions were distilled to dryness and treated with 4 N hydrogen chloride in ethyl acetate to remove tert. Boc-protecting group. The resulting hydrochloride salt is converted to the free base by absorption on a highly acidic ion exchange resin and eluted with a 2X-pyridine solution in water. The product rich fractions were freeze-dried to give N, N-1-carboxy-3-phenylpropyl] -L-lysyl-L-proline as a white fluffy substance. The NMR spectrum is consistent with the structure. The mass spectrum shows a molecular ion at 549 for the disilylated species. Chromatography results in the desired isomer.

Example 58. L1- (1-carboxy-3-phenylpropyl) -L-3-fluoroalanil-L-proline.

To a solution of D-3-fluoroalanine (420 mg) in 4 ml of acetone (1: 1) was added triethylamine (590 mg) and 2-tert-butoxycarbonyloxyimino-2-phenylacetonitrile (1.060 g). The mixture was stirred for two and a half hours. Cold 5X aqueous aqueous potassium bicarbonate was added and the mixture was extracted with ethyl acetate. The aqueous phase was acidified with 1 N hydrochloric acid and extracted with ethyl acetate. The last extract was washed with saturated aqueous sodium chloride solution, dried over sodium sulfate and concentrated to dryness to give L-tert. Boc-3-fluoroalanine (800 mg / mp). 91-93 ° C.

When stirred, a solution of the above product (800 mg) and proline benzyl ester (1.5 g) in methylene chloride (ml) at 0 ° C was added dicyclohexylcarbodiimide (845 mg) in methylene chloride (6 ml) and the mixture was added. maintained at 0 ° C for 2 hours and at temperature

- 51 20 ° C for 18 hours. The mixture was filtered, the precipitate was washed with methylene chloride, the filtrate and washes were collected and extracted with cold 1 N hydrochloric acid, cold 5-potassium aqueous potassium bicarbonate, saturated aqueous sodium chloride. , dried over sodium sulfate and concentrated to dryness. Dry column chromatography on silica gel H eluted with a 6% solution of acetone in chloroform to give the pure protected dipeptide.

The tert.-Boc group was removed by treatment with a 4 N solution of hydrogen chloride in ethyl acetate (8 ml) at 0 ° C for one hour. Ether (about 20 ml) was added and the precipitate formed

1-3-Fluoroalanil-L-proline benzyl ester, hydrochloride (450 mg) was collected by filtration (m.p. Mp 158-161 ° C. After hydrogenation in 6 ml of water and 2 ml of ethanol over 60 mg of 10H-palladium / carbon at a pressure of one atmosphere and a temperature of 20 ° C for 90 minutes, followed by filtration and concentration to dryness, L-3-fluoroalanil-L is obtained. -proline hydrochloride (330 mg). The mass spectrum shows a molecular ion at 348 m / e for the dimethylsilylated species.

To a mixture of 4-phenyl-2-oxoic acid (375 mg) and L-fluoroalanil-L -proline hydrochloride (100 mg) in 3 ml of water (pH adjusted to 7 with sodium hydrochloride) was added sodium cyanoborohydride (80 mg). . The mixture was stirred for 20 hours and then treated according to the method described in Example 24. The mass spectrum of the product purified by L H-20 showed a molecular ion at 510 m / e for the dimethylsilylated species. Silica gel thin layer chromatography showed a spot with = 0.7 using an ethyl acetate / acetic acid / nbutanol / water system in a 1: 1: 1: 1 ratio.

- 52 The diastereoisomers were separated by chromatography on XAD -2 resin according to the method described in Example 25.

L1- (1'-ethoxycarbonyl-3-phenylpropyl) -L-3-fluorine or lL-proline was prepared by the method described in Example 26.

Example 59 L1- (1-ethoxycarbonyl-3-phenylpropyl) -L-alanylL -3,4-dehydropropion.

In the method described in Example 26, the L-alanyl-L -3,4-dehydroproline obtained in Example 37 is converted to 11- (1) -ethoxycarbonyl-

3-phenylpropyl] -L-alanyl-L -3,4-dehydroproline in the form of a two-component diastereomeric mixture. Thin layer chromatography on silica gel showed two spots of cR ^ 0.79 (extra small) at two developments in a 4: 1: 1 ratio of n-butanol / water / acetic acid. The mass spectrum shows molecular ion at 518 m / e for the dimethylmethyl silylated species.

EXAMPLE 60 N- (1/57-methoxycarbonyl-2- (1H-imidazol-4-yl) ethyl] -Galanyl-L-proline.

In the method described in Example 16, the L-histidine methyl ester was condensed with pyrrovoyl-L-proline in the presence of sodium cyanoborohydride to give N- (1-methoxycarbonyl-2 / 1H-imidazol-4-yl) ethyl / -1 > 1chalanyl-L -proline. The NMR spectrum in deuterated water showed imidazole protons at 8.6 and direct 7.3. The protons adjacent to imidazole and the methyl ester protons appear at 3.7, and the olanil methyls at 1.1 to 1.38.

Example 61 N- [1- (N) -carboxy-2- (1H-imidazol-4-yl) ethyl] -L-alanyl-L-proline.

In a method similar to that described in Example 17, the product obtained in Example 60 is characterized to yield the expected diacid. The NMR spectrum in deuterated water shows imidazole protons at 7.2 and 8.5; and ayanine methyl at

1.25 ί.

Example 62 N- (R) -ethoxycarbonyl-5-aminopentyl (-1), L alanyl-L-proline.

A solution of e-benzyloxycarbonyl-L-lysine ethyl ester hydrochloride (2.94 g) in water (10 ml) was made basic with 15 ml of saturated potassium bicarbonate solution and extracted with methylene chloride. The extract was dried over magnesium sulfate and concentrated to dryness. The residue of N-benzyl oxycarbonyl-L-lysine ethyl ester was dissolved in THF (20 mL) and pyruvoylproline (555 mg) and molecular sieve No. 4A (1.0 g) were added. The mixture was stirred at room temperature for 4 hours. A solution of sodium cyanoborohydride (630 mg) in 1 ml of methanol was added over 2 hours and the mixture was stirred overnight. It was then filtered, concentrated to dryness, and the residue partitioned between water (10 ml) and methylene chloride (15 ml). The aqueous phase was absorbed on a highly acidic ion exchange resin and eluted with 4X-pyridine solution in water to give 470 mg of N- (I) (5) ethoxycarbonyl-5-benzyloxycarbonylaminopentyl) -B, L-alanylL-proline. The protecting group is removed by hydrogenation in ethanol-water (1 to 1) over 10% palladium / charcoal at 40 psi pounds per square inch absolute pressure. The mixture was filtered and the filtrate was evaporated to dryness. The residue was dissolved in methanol and chromatographed on a column of LH-20 to give the desired N- (R) -ethoxycarbonyl-5-aminopentyl) -L, L-alanyl-L-proline. The mass spectrum of the silylated material shows a peak at 559 m / e for the trisilylated derivative and a peak at 487 m / e for the disilylated derivative. The NMR spectrum in deuterated water confirms the structure.

Example 54. N- [1- (5) carboxy-5-aminopentyl] -L-alanyl-L proline.

The N- (1/5) -ethoxycarbonyl-5-benzyloxycarbonyl-aminolentyl (-)), obtained in Example 62, was treated with 0.1 M sodium hydroxide solution at room temperature overnight. After absorption of the product on a highly acidic ion exchange resin, the product was eluted with a 4% solution of pyridine in water to give 1- (1 / $) - carboxy-

5-benzyloxycarbonylaminopentyl (-L, L-alanyl-L-proline, which, by thin layer chromatography, shows a single spot with R 0.4, (10: 4: 3: 1) butanol-water-pyridine-acetic acid system. In a method similar to that described in Example 62, the protecting group is removed by hydrogenation to give N- (R) carboxy-5-aminopentyl--5, L-alanyl-L-proline. The mass spectrum of the trimethylsilylated product is consistent with the structure and has a mass peak at 531 m / e. Chromatography afforded the desired isomer.

Example 64. N-1- (L-carboxy-6-aminohexyl) -L-alanyl-L proline.

Benzyl 2-oxo-7-phthalimidoheptanoate, obtained after alkylation of benzyl 1,3-dithian-2-carboxylate with 5-phthalimidopentyl bromide and after oxidative conversion to the ketone with N-bromosuccinimide is condensed with L-alanyl-L -proline of excess cyanoborohydride. The condensation product N- (1-benzylsilicarbonyl-6-phthalimidohexyl) -L-alanyl-L proline (390 mg) in 25 ml of a 50% ethanol-water mixture was hydrogenated at 40 psi / abs. pressure in pounds per square inch / on southeast palladium / charcoal. Removal of the solvent and catalyst gave N- (1-carboxy-6-phthalimidohexyl) - 55 L-alanyl-L -proline (320 mg), which has the expected spectral and chromatographic characteristics. Part of the intermediate obtained above (152 mg) in 2 ml of ethanol was refluxed * with hydrazine (32 mg) for an hour and a half. The phthalhydrazide was removed by filtration; the ethanol was distilled in vacuo and the residue was absorbed on a highly acidic ion exchange resin. Elution with 2N aqueous pyridine solution and freeze-drying afforded the desired N- (1-carboxy-

6-aminohexyl (-L-alanyl-L-proline (58 mg)). The spectral data are consistent with the structure. The mass spectrum shows a peak at 311 for the molecular ion minus water (329-18).

Example 65 N- (1-benzyloxycarbonyl-6-aminohexyl) -L alanyl-L proline.

Carrying out the hydrolysis according to the procedure described in Example 64, b-1- (L-benzyloxycarbonyl-6-phthalimidohexyl) -L-alanyl-L-proline produced a mixture from which N- (L-benzyloxycarbonyl-6-aminohexyl) - alanyl-L -proline. Thin layer chromatography on silica gel eluting with a mixture of 50 butanol, 41 acetic acid and 30 water gave a spot with R _F 0.32.

Example 66 N- (1-carboxy-2-phenoxyethyl) -L-alynyl-L · proline.

Suspension of phenoxypyruvic acid (1.8 g), obtained by condensation of ethyl phenoxyacetate with diethyl oxalate, followed by catalytic acid hydrolysis and decarboxylation, and Lalanyl-L-proline (0.37G*) in 10 ml of water, adjusted to 7 with pH adjusted to 7 diluted sodium hydroxide, treated with sodium cyanoborohydride (0.18 g) and stirred at room temperature for five days. On the second and third days, additional keto acid (0.9 g) and sodium cyanoborohydride (0.18 g) were added.

56 The product is absorbed on a highly acidic ion exchange resin, eluted with a 2% solution of pyridine in water, and after freeze-drying, 0.5 g of N- (1-carboxy-2-phenoxyethyl) -L-alanyl-L is obtained. proline. The NMR spectrum is consistent with the structure. The mass spectrum shows a peak at 479 for the silyl molecule ion minus methyl (494-15).

Example 67. N- (1-Ethoxycarbonyl-2-phenoxyethyl) -L-alanylL-proline.

By reaction of ethyl phenoxy pyruvate obtained by acid-catalyzed esterification with the corresponding acid and L-alanyl-L-proline with sodium cyanoborohydride in ethanol solution and isolation of the product as described in Example 66, b1 can be obtained - (1-Ethoxycarbonyl-2-phenoxyethyl) L-alanyl-L-proline.

Example 68. b [- (1-carboxy-2-phenylthioethyl) -L-alanyl-L proline.

Mixture of phenylthiopyrogen-dehydrated nitrate (1.96 g) obtained by condensation of ethyl phenylthioacetate with diethyloxalate and then by acidic hydrolysis and decarboxylation, and L-alanyl-L-proline (0.37 g) in 10 ml of water, diluted to 7.0 with a dilution of 7.0 with dilution of sodium hydroxide is treated with sodium cyanoborohydride (0.18 g) in 2 ml of water. After stirring overnight at room temperature, the product was absorbed onto a highly acidic ion exchange resin and eluted with a 2 * pyridine solution in water to give 0.36 g of 1 '[- / 1-narboxy-

2-Phenylthioethyl N-alanyl-L-proline. NMR and mass spectra confirm the desired structure. A mass peak at 348 indicates the molecular ion (366) minus water (18).

57 Example 69 N- (1'-ethoxycarbonyl-2-phenylthioethyl) -L-alanylL-proline.

By reacting the ethyl phenylthiopyrate obtained from the acid by esterification, and L-alanyl-L-proline with sodium cyanoborohydride in ethanol solution as described in Example 68 and by isolating the product in the manner described therein, 1-ethoxycarbonyl-2-phenylthioethyl) -L alanyl-L-proline.

Example 70. N- (1-Ethoxycarbonyl-3-p-chlorophenylpropyl) -alkanyl-L-proline.

A solution of ethyl 4-p-chlorophenyl-2-oxobutyrate obtained from the acid by esterification with ethanol in tetrachloromethane at reflux and L-alanyl-L-proline in ethanol is treated with excess sodium cyanoborohydride at room temperature and digested at room temperature until the reaction is complete. The ethanol was removed in vacuo and the product was absorbed on a highly acidic ion exchange resin. By eluting with a 2X-ene solution of pyridine in water, N- (1-hydroxy-carbonyl-3-p-chlorophenylpropyl) -L-alanyl-L-proline can be obtained.

Example 71. 1- (1-Ethoxycarbonyl-2- (3-idolyl) ethyl) -alkanyl-L-proline.

In the manner described in Example 26, the indole-3-pyruvic acid ethyl ester was condensed with L-alanyl-L proline in a 1: 1 mixture of ethanol and water using tarien cyanoborohydride. Isolation on Dowex 50 yields a mixture of N- (1-ethoxycarbonyl-2/3-indolyl) ethyl-L-alanyl-L-proline isomers in the manner described. The NMR spectrum showed absorption at 7.0 ί (5H), 4.2 (5H), 3.4 Г (2H), 2.9 672H (1.9), 6H (1.2), and 1.2 (bN).

Example 58. N- (1-Ethoxycarbonyl-2-p-aminomethylphenylethyl) -L-alanyl-L · -proline.

Ethyl 2-oxo-3-p-cyanophenylpropanoate obtained by reacting p-cyanobenzyl bromide with ethyl 1,3-dithian-2-carboxylate and subsequent oxidative hydrolysis by the method described by Eliel and Hartmann, J. Org, Chem., 37, 505 (1972) was condensed with L-alanyl-L-proline and the product was purified by the method described in Example 1. Hydrogenation of the resulting mixture of N- (1-ethoxycarbonyl-2-p-cyanophenylethyl) isomers ~ L-alanyl-L-proline in ethanol solution containing hydrogen chloride and palladium on carbon as catalyst. Distillation of the solvent and excess hydrogen chloride in vacuo, pouring with ethanol and concentrating to dryness to obtain the hydrochlorides of the mixtures of diastereoisomers of the desired compound.

Example 73. L1- (1) Carboxy-2-p-aminomethylphenylethyl) -L alanyl-L-proline.

A sample of N- (1-ethoxycarbonyl-2-p-cyanophenylethyl) -cylanyl-L-proline prepared above in Example 72 was treated with one equivalent of sodium hydroxide in a mixture of methanol and water as a solvent at room temperature for one night. Distillation of the solvents in vacuo afforded the sodium salts of the mixture of isomers of N- (Lcarboxy-2-cyanophenylethyl) -L-alanyl-L-proline. This mixture is hydrogenated in a solution of ethanol hydrochloric acid and developed in the manner described in Example 72 to give the hydrochlorides of the mixture of diastereoisomers of the desired product.

Example 74 1 N- (1-ethoxycarbonyl-2/5) -amino-3-phenylpropyl / D, 1 galanyl-L-proline.

To a mixture of N-phthaloyl-L-2-amino-3-phenylpropionaldehyde

59 (Petf.rson et al., J. Am. Chem. Soc., 79, 1398 (1957), (2.18 g) and potassium metabisulphide (0.87 g) in a 1: 1 mixture of water and methanol. sodium cyanide (0.55 g) with vigorous stirring, stirring was continued for 90 minutes, then the mixture was diluted with ethyl acetate and filtered, the organic layer was washed with water and dried over magnesium sulfate, the solvent was removed in vacuo to give N-phthaliol -

3-amino-4-phenyl-2-hydroxybutyronitrile, thin layer chromatography with ethyl acetate / hexane in 1: 1 ratio gave a spot with RF 0.5.

A solution of this product in absolute ethanol is prepared, which is saturated with ammonia and left to stand for 6 days at room temperature. The solvent was removed, the residue was taken up in dioxane / concentrated hydrochloric acid in a 1: 1 ratio, warmed to 70 ° C and held at this temperature for 20 hours. Evaporation of the solvent to dry suspension of the residue with warm water, filtration and purification on a highly acidic ion exchange resin in the usual way to give / 2R, S; 35 / -2-amino-4-phenyl-3-phthaloylaminobutanoic acid. Dissolving the acid in anhydrous ethanol, passing the anhydrous hydrogen chloride to saturation and maintaining the mixture at room temperature for 16 hours. Removal of the solvent in vacuo to yield the amino acid hydrochloride of the amino acid. Condensation of this ethyl 2-amino-4-phenyl-3-phthalylamino-butanoate with pyruvoyl-L-proline sodium cyanoborohydride, as described in Example 16, to give L1- [1- ethoxycarbonyl-3-phenyl-

2-Phthaloylaminopropyl-L, L-alanyl-L-proline in the form of a mixture of isomers. The reflux of this material in ethanol with one envalvate hydrazine is cooled for an hour and a half, filtering the phthalhydrazide and isolating the desired product from the resulting mixture by chromatographic methods to give N- (1-carbethoxy-2/5) - amino-3-phenylpropyl-P, L alanyl-L-proline. The NMR spectrum at 300 MH + has peaks at 7.3 Г (5H), 3.2 and 4.4? (3H), 2.75 </ (2H), 2.0 ί (4H) and 1.1 8 (M).

Example 75 N- (1-carboxy-2/5) -amino-3-phenylpropyl-L, L alanyl-L-proline.

The 2-amino-4-phenyl-3- (5 / -3-phthaloylaminobutanoic acid) obtained in Example 74 was condensed with pyruvoyl L-proline using sodium cyanoborohydride as described in Example 16 to give L1- [1] carboxy- 3-phenyl-2-phthaloylamino-1, L alanyl-L-proline as a mixture of isomers. This material was refluxed in ethanol together with one equivalent of hydrazine for an hour and a half, cooling, filtering the phthalohydrazide, isolating the desired product by chromatographic procedures to afford the title compound. Thin layer chromatography on silica gel eluting with ethyl acetate to 5 pyridine to 1 acetic acid to 3 water gave a net spot with R _F 0.15.

Example 76 N- (1-carboxy-2/5) -benzoylamino-3-phenylpropyl) D, L-alanyl-L-proline.

A solution of N-phthaloyl-3-amino-4-phenyl-2-hydroxy butyronitrile, obtained in Example 74, was allowed to stand in anhydrous ammonia-saturated ethanol for 3 days at room temperature. The solvent was removed in vacuo and the residue was refluxed for 6 hours in concentrated hydrochloric acid. Evaporate to dryness, purify the residue in a column of Dowex 50 (H +) ion exchange resin, eluting

- 61 in succession with water / methanol / 10: 1, water / pyridine / 50: 1 and finally with 0.5 M ammonium hydroxide solution. From this last eluent was isolated after concentration to the dry desired 2,3-diamino-4-phenyl propionic acid. A solution of a copper complex of this amino acid and benzoylate is prepared at the amino group in the third position with benzoyl chloride under alkaline conditions and by the method described by R · Hoeske et al., J, Am. Let's sleep. Soc. ,> 73, 5883 (1956). Cleavage of the copper complex with hydrogen sulfide and treatment of the mixture in the manner described to give 2amino-3- (S) -benzoylamino-4-phenyl butyric acid. Condensation of this intermediate with pyruvoyl 2-proline using sodium cyanoborohydride in the manner described in Example 16 to give the desired N 1 - [(- carboxy-2/5 / -benzoylamino-3-phenylpropyl) -B _L _ _alanyl -L -proline in the form of a mixture of isomers that can be separated by chromatographic methods, as appropriate.The mass spectral analysis shows a peak at 596 m / e for the disilylated molecular ion minus IS, the NMR spectrum shows the absorption for two phenyl at 7.0-7.5 $ and for diastereomeric methyl at II cP.

Example 77. N- (1-Ethoxycarbonyl-2- (S) -benzoylamino-3-phenyl-propyl) -L-alanyl-L-proline.

By treating 2-amino-3-benzoylamino-4-phenyl butyric acid obtained in Example 76 with saturated hydrogen chloride solution in absolute ethanol for 4 h and subsequent distillation of the solvent in vacuo, ethyl can be obtained

2-amino-3-benzoylomino-4-phenyl butyrate hydrochloride. By condensation of this intermediate with pyruvoyl -L-proline using sodium cyano borohydride in the manner described in Example 16, and isolating the product in the manner described therein,

- 62 the title compound was obtained.

Example 78 N- (2-amino-1-carboxy-4-methylpentyl) -1, L alanyl-L-proline.

A solution of 0.731 g of trans-3-amino-4- (2-methylpropyl) -2-azetidinone, obtained by the addition of chlorosulfonyl isocyanate to 4-methyl-1'-pentene; the resulting lactam is protected as tert. butyldimethylsilyl derivative and then treated with lithium diisopropylamide, followed by tosyl azide and chloromethylsilane; acidification and chromatography on silica gel gave tross-3-azido-4- (2-methylpropyl) -2-azetidinone, which was hydrogenated with 10% palladium / carbon in ethanol to the corresponding amino derivative and 4.58 g benzylpyruvate in 20 ml absolute ethanol, containing 10 g of the powdery molecular sieve 4A was treated dropwise with a solution of sodium cyanoborohydride (0.65 g) in 8 ml of absolute ethanol with stirring until the reaction was complete. The reaction mixture was filtered and the filtrate was concentrated. The residue was dissolved in 50 ml of water and acidified with 1 N hydrochloric acid solution to pH3. The mixture was basified with 10N sodium carbonate solution, the aqueous solution was saturated with sodium chloride and extracted with ethyl acetate (5x40 ml). The combined organic layers were dried over sodium sulfate and concentrated to give an oil (4.94 g). Chromatography on silica gel (ethyl acetate) gave I. N g of product. NMR and mass spectrograms are consistent with the structure of N- (trans-4- (2-methylpropyl) -2-oxo-3-azetidinyl) -1, L-alanine benzyl ester. The debenzylation is carried out by hydrogenation with 10-palladium / charcoal in a mixture of ethanol / water at a ratio of 2: 1. Cold solution at 0 ° C acid (428 mg)

- 63 and L-proline tert-butyl ester (377 mg) is 5 ml of dimethylformamide treated with a solution of diphenylphosphoryl azide (605 mg) in 5 ml of dimethylformamide and then with a solution of triethylamine (223 mg in 5 ml of dimethylformamide). for 20 minutes. After three hours, the ice bath was removed and the reaction mixture was stirred at room temperature overnight. Ethyl acetate (100 ml) was added and the resulting solution was washed with water (2x40 ml), 5X sodium carbonate solution (3 x 30 ml) and water (1 x 50 ml) before drying with sodium sulfate. Concentration gave an oil of 0.78 g, whose NMR and mass spectra were consistent with the structure of N- (trans-4-).

2-methylpropyl (-2-oxo-3-azetidinyl) -0,11-alanyl-L · -proline butyl ester. The crude product was dissolved in 25 ml of trifluoroacetic acid at 0 ° C. The reaction mixture was stirred at 0 ° C for 20 min and then at room temperature for two and a half hours. The reaction mixture was concentrated to dryness and the residue was treated with 1 N sodium hydroxide solution (30 ml) for four and a half hours at room temperature. The alkaline mixture was slowly added to the highly acidic ion-exchange ion ^, and the product was recovered with a 2% solution of pyridine V water. Drying by means of

S 'mrazyavane affords ^da g of N- / 2-amino-1-carboxy-

4-methylpentyl) -P, L-alayl-proline consisting of four

V diastereomers (S, S, S, o, R, $; R, R, R, 5; R, R, 5 »S '; / which can be separated by chromatography. The NMR and mass spectra are consistent with the structure. The NMR spectrum showed multiplets centered at 4.5, 3.85, 2.3, 1.79 and at 1.16 ppm. The mass spectrogram showed a peak at 458 m / e (molecular ion of disilylated material minus 15).

BAD ORIGINAL $

- 64 Example 79. N- / 2-Ethoxycarbonyl-4-methylpentyl / P, L-alanyl-L · -proline.

An intermediate obtained in Example 78, N- / trans-

4- (2-methylpropyl) -2-oxo-3-azetidinyl) -B, L-alanine (125 mg) can be condensed with L-proline benzyl ester hydrochloride (167 mg) in the presence of diphenylphosphoryl azide (191 mg) and triethylamine (140 mg) in dimethylformamide solution to give 217 mg of N- (trans-4/2-methylpropyl) -2-oxo-3-azetidionyl) -P, L-alanyl-L-propenzyl ester. The benzyl protecting group can be removed by hydrogenation and the N-lactam can be opened with anhydrous Notrium ethoxide in ethanol to give L 1- (2-amino-1-ethoxycarbonyl-4-methyl-pentyl / D). L-alanyl-L · -proline.

Example 80. N- / 2-Benzamido-1'-Carboxy-4-methylpentyl / P, L-alanyl-L-proline.

A solution of N- (2-amino-1'Carboxy-4-methylpentyl (-P, L alanyl-L-proline) prepared in the manner described in Example 78 was treated with benzoyl chloride in aqueous alkaline to give M- (2-Benzamido-1-carboxy-4-methylpentyl) -P, L alanyl-L-proline. The masseter of the distilled material has peaks at 562 (M ⁺ -15), 460, 389 (base peak), 364, 242 and 174 m / e.

Example 81 b1- (2-Benzamido-1-ethoxycarbonyl-4-methylpentyl) P, L-alanyl-L-proline.

When treating a solution of N- [2-amino-1 · -ethoxycarbonyl

4-methylpentyl (-P, L-alanyl-L-proline), prepared in the manner described in Example 79, can give N- (2-benzamido-1-ethoxycarbonyl-4-methyl-65 in an organic solvent with benzoyl chloride). pentyl / N, L-alanyl-L-proline.

Example 82. N- tZ - (1-ethoxycarbonyl-3-phenylpropyl) -L alginyl- L -proline.

In the method described in Example 37, N - ^ - tert is non-condensed. Boc-N-W-nitro-L-arginine N-hydroxy succinimide ester with L -proline in a mixture of dioxane and water, the N- < Z-tert.-Boc protecting group was removed with trifluoroacetic acid and the dipeptide was isolated as described. The coupling of the product obtained with ethyl 2-oxo-4-phenylbutyrate and isolation in the manner described in Example 26 results in a material having a U-nitrogen of arginine still protected by a nitro group. Removal of the protecting group by catalytic hydrogenation in ethanol-acetic acid over palladium / on carbon as a catalyst at room temperature and 40 pounds of hydrogen pressure. Filtration of the catalyst and distillation of the solvents in vacuo afforded a mixture of isomers of N- / - / - ethoxycarbonyl (-3-phenylpropyl) -L-arginyl-L-proline.

Example 83. N-Z, - (1-carboxy-3-phenylpropyl) -B, L-homolizyl-L-proline.

The condensation of N - N - benzyloxycarbonyl-N, N, -3 ° butoxycarbonyl-D, L-homolysin (obtained from hemolysin through a copper complex with L-proline-3 ^O butyl ester) can be accomplished by using definylphosphorylazide. The 3 ° butyl groups can then be removed with trifluoroacetic acid, and the product UJ-benzyloxycarbonyl-D, Xhhomosolyl-L-proline can be reacted with 2-oxo-4-phenylbutyric acid and sodium cyanoborohydride The condensation product can be catenitized but hydrogenation to give N- [S] -carboxy-3-phenylpropyl] -P, L-homolizyl-L-proline.

- 66 EXAMPLE 84 N-Z - (containing ester-sicarbonyl-3-phenylpropyl) -B,

L-homolysil-L · -proline.

The intermediate described in Example 83, N- (S) -benzyloxycarbonyl-D, L-homolizyl-L-proline, can be reacted with ethyl 2-oxo-4-phenylbutyrate and sodium cyanoborohydride. The condensation product can then be hydrogenated over palladium / charcoal to give N- (R) - (1-ethoxycarbonyl).

3-phenylpropyl-P, L-homolysyl-L-proline.

Example 85. N-5 N - (T-carboxy-3-phenylpropyl) - N - aminoR, L-alanyl-L-proline.

Under alkaline conditions, B, N, N, β-diominopropionic acid is reacted with excess benzyloxycarbonyl chloride to give the acidification of D, L - N- (bis) benzyloxycarbonylamino (propionic acid); 123.5-124 ° C. Phosphorous pentachloride was added to the chloroform solution containing this product to give the development of D, L-

4- (benzylsulfonylaminomethyl) -oxazolidin-2,5-dione. L -proline tert solution. butyl ester in methylene chloride is added to N-carboxylic anhydride in tetrahydrofuran at -60 ° C. After standing overnight under freezing conditions, the mixture was evaporated to dryness. The crude product is obtained. Trifluoroacetic acid, which effectively cleaves the tert-butyl ester group for two hours at room temperature, resulting in a mixture of L-proline and N-benzyloxycarbonylamino-L, L-alanyl-L-proline. Gel chromium filtration on column b-H-20 gave a pure dipeptide.

2-Oxo-4-phenyl-fat oil condensed and benzyloxy III UP

BAD ORIGINAL D ^ka P ^{Bonnie /} amine ° -1), L-alanyl-L-proline in the presence of sodium ^{1 11} '..... cyanoborohydride. Removal of the protecting group from the resulting product yields N- (1-carboxy-3-phenyl-propyl) -N-amino-D, L-alanyl-L> -proline. NMR spectrum in deuterated. ------ „„ ntltltli pptptni poi 7.4 cP. Metin protons

- 67 at 4.2 ° and multiplets at 3.7, 3.2, 2.9 and 2.1

Example 86 N-N- (N-ethoxycarbonyl-3-phenylpropyl) -L amino-D, L-alanyl-L-proline.

A solution of β-benzyloxycarbonylamino-P, L-alanyl-L proline obtained in the method described in Example 85, and ethyl 2-oxo-4-phenylbutyrate are condensed in ethanol with sodium cyanoborohydride. The protecting group can be removed from the product by catalytic hydrogenation to give N - (N - (heteroxycarbonyl-3-phenylpropyl) - N -amino-N, L-alanyl-L-proline.

Example 87. N- [1- (5) -Ethoxycarbonyl-3-phenylpropyl-O, L p-aminomethylphenylalanil-L-proline.

Ethyl 2-oxo-3-p-cyanophenylpropionate prepared as an example

72, was hydrolyzed by stirring with 5 * sodium hydroxide solution at room temperature overnight. The reaction mixture was washed with ether, acidified the aqueous layer to pH 2 with concentrated hydrochloric acid, extracted the product into a mixture of ether and ethyl acetate and the solvent removed to give 2-oxo-3-p-cyanophenylpropionic acid. Following the reductive coupling of the acid with L-homophenylalanine ethyl ester in the presence of sodium cyanoborohydride and further purification as described in Example 13, a mixture of N- (I- (5) carbotexy-3-phenyl-propyl) diastereomers was obtained. -P, L-p-cyanophenylalanil-L-proline as benzyl ester. Hydrogenation of this intermediate in ethanol containing hydrogen chloride on palladium / charcoal in the manner described in Example 72 and treatment of the mixture in the manner described in the same example gave the desired product in the form of a mixture of diastereoisomers.

- 68 Example 88. N-Z - (1/5) carboxy-3-phenylpropyl) -P, L-amino-methylphenylalanil-L-proline.

The N- (1- (S) -ethoxycarbonyl-3-phenylpropyl) -P, L - p-cyanophenylalanil-L-proline benzyl ester obtained in Example 87 was hydrolyzed by treatment with two equivalents of sodium hydroxide in a mixture of methanol and water at room temperature overnight. The solvent was removed from the reaction mixture by distillation in vacuo and the residue was hydrogenated in ethanol hydrochloric acid as described in Example 72. After treating the mixture further, a mixture of the diastereoisomers of the desired product is obtained in the manner described in the same example.

Example 89. - (1-Ethoxycarbonyl-3-phenylpropyl) -Macetyl-L-lysyl-L-proline.

In the method described in Example 26, ethyl 2-oxo-4-phenylbutyrate is attached to N, N-acetyl-L-lysyl-L-proline in a solvent mixture of ethanol and water in the presence of sodium cyanoborohydride. Isolation by Dowex 50 as described above and freeze-drying of the product-rich fractions yielded a mixture of N- / Tetoxycarbonyl-3-phenylpropyl - / - 11- isomers. -acetyl-L-lysyl-L -proline. The mass spectrum of the trisilylated material peaks at 663 (M *), 648 (M + - 15) and at 546 m / e.

Example 90. N-N- (L-ethoxycarbonyl) -L-histidyl-L-proline.

As described in Example 26, ethyl 2-oxo-4-phenylbutyrate is reacted with L-histidyl-L-proline in the presence of sodium cyanoborohydride. Purification in the manner described gives the mixture of diastereoisomers of M-X- (1-ethoxycarbonyl) -L-histidyl-L-proline.

- 69 Example 91. N - [- (ethoxycarbonyl-3 - / - naphthylpropyl) -L lysyl-L-proline.

Ethyl N - naphthyl -2-oxobutyrate obtained by alkylation of ethyl 1,3-dithian-2-carboxylate with 2 - / - naphthalyl bromide and subsequent conversion with ketone with N-bromosuccinimide to aqueous acetone is condensed with N - 3 ° Butoxycarbonyl-L-lysyl-L-proline in ethanol in the presence of sodium cyanoborohydride and molecular sieves The product is then absorbed onto a highly acidic ion exchange resin and eluted with a 2% solution of pyridine in water. The cleavage of tert.-Boc protecting group can be accomplished by treatment with a 4.0 N hydrochloric acid solution in ethyl acetate to give N - (ethoxycarbonyl-3 -) - naphthylpropyl (L-lysyl-L-proline).

B. G'1- _! N- (1'-carboxy-3 - / - naphthylpropyl) -L-lysyl-L-proline.

The 4-N-naphthyl-2-oxobutyric acid obtained from the ester by hydrolysis is suspended in water and the pH of the medium is adjusted to 7 using a dilute sodium hydroxide solution and then freeze-dried. The residue was treated with -30-butoxycarbonyl-L-lysyl-L-proline in the manner described in part A. to give L1 - [- (L-carboxy-3) -naphthylpropyl] N-lysyl-L-proline.

Example 92. N - N - (1/5) carboxy-3-p-chlorophenylpropyl) -Elysyl-L-proline.

Solution Ha-3 ° -6yTOKCHKap6oHnn-L-lysyl-L-proline (0.36 g) and 4-p-chlorophenyl-2-oxobutyric acid (1.1 g) in 5 ml of water after adjusting the pH to 7 seconds using a dilute sodium hydroxide solution, treat with 0.07 g of cyanoborohydride in 1 ml of water for several hours. After stirring the mixture at room temperature overnight, the product was stirred

70 was absorbed on a highly acidic ion exchange resin and eluted with a 2 ^ pyridine solution in water to give 0.058 g of product. The NMR spectrum indicated that the tert.-Boc-protecting group had not been completely cleaved. The product was treated with 4.5 N HCl in ethyl acetate, followed by isolation by ion exchange resin to afford 0.048 g of N- _</ - / 1gkarboksi-

3-p-chlorophenylpropyl N-L-lysyl-L-proline. The NMR and mass spectra are consistent with the structure. The peak at 584 corresponds to the silylated molecular ion. Chromatography yields the desired isomer.

Example 93 N-, N- (1-ethoxycarbonyl-3-p-chlorophenyl) -L-lysylL-proline.

Condensation of ethyl 4-p-chlorophenyl-2-oxobutyrate and N-3-butoxycarbonyl-L-lysyl-L-proline in ethanol solution in the presence of excess sodium cyanoborohydride and isolation of the product as described in Example 92 is obtained N - [(1) getoxycarbonyl-3-p-chlorophenylpropyl) -L-lysyl-L-proline.

Example 94 Ν-ο (- / Ι carboxy-3- / 3,4-dichlorophenyl) -propyl / L-lysyl-L-proline.

A solution of 4- (3,4-dichlorophenyl) -2-oxobutyric acid, obtained from dichlorodihydrocinnamate ester by condensation with ethoxalate and subsequent acid hydrolysis and partial decarboxylation, was treated in water according to the method described in Example 92 to give N - / - (L) carboxy-3- (8,4-dichlorophenyl) propyl-L-lysyl-L-proline. The NMR spectrum in deuterated water showed aromatic hydrogen at 7.30 lb. wide singlet at 4.35 δ and wide multiplets at 3.53, 3.00, 2.68, 1.98 and 1.65 §. The mass spectrum shows a peak at 455 m / e, which corresponds to a loss!

- 71 of water of the relevant structure. A large M + 2/457 m / e / isotope indicates the presence of 2 CI.

Example 95 N - (- (1/6) -carboxy-3- (3-indolyl) propyl) -L lysyl-L-proline.

2-Oxo-4- (3-indolyl) -butyric acid is obtained from homotryptophan by the method described by Weygand et al., Ann. 658,128 (1962). This acid was condensed with e-tert. Boc-L-lysyl-L-proline in the presence of sodium cyanoborohydride in the manner described in Example 54 to give a crude mixture of diastereoisomers of N - (1) Narboxy-3- (3). -indolyl / prolyl-L1-6-tert-Boc-L-lysyl-L-proline. The mass spectrum of the trisilylated material peaks at 732 (M *), 717 (M * -15), 615, 660, 645 m / e. The lysine side chain protecting group is cleaved by treatment with 4 N hydrochloride in ethyl acetate and purification on a highly acidic ion exchange resin in the manner described in Example 54 to give the desired product. Chromatography can give the desired isomer.

Example 96. N ~ t - [(6-aNMHO-Ir narboxyethyl) -L-lysyl-Lproline.

Benzyl 2-oxo-7-phthalimidoheptanoate and 6-3 ° -butoxycarbonylL-lysyl-L-proline were condensed with excess sodium cyanoborohydride in ethanol solution to give N-N- (1-benzyloxycarbonyl-6-phthalimidox-1-phthalimidoxyl-6-phthalimidoxyl-6-phthalimidoxyl-6-phthalimidoxyl-6-phthalimidoxyl-6-phthalimidoxyl-6-phthalimidoxyl-6-phthalimidoxyl-6-phthalimidox. ^N -3 ^O -butoxycarbonyl-L lysyl-L-proline. The benzyl group can then be removed by hydrogenation over palladium / carbon, and the tert.The Boc protecting group can be cleaved with 4.5 N hydrogen chloride in ethyl acetate, and the phthalimide group by treatment with hydrazine to give M- _O / - / 6- amino-L-carboxyhexyl / -L-lysyl-L · -proline.

Example 97. N- _O- (6-amino-L-benzyloxycarbonylhexyl) - 72 L-lysyl-L-proline.

By treating N- (N -benzyloxycarbonyl-6-phthalimidohexyl) -N-N-3-butoxycarbonyl-L-lysyl-L-proline prepared by the method described in Example 96 with 4.0 N hydrogen chloride in ethyl acetate and then with one hydrazine equivalent at reflux 8 ethanol to give a mixture from which N- (S-amino-T-benzyloxycarbonylhexyl) -L-lysyl-L-proline can be isolated.

EXAMPLE 98 N- [5- (5-amino-1- (S) -carboxypentyl) -N-lysyl] -L-proline.

Benzyl 2-oxo-6-phthalimidohexanoate was treated in the manner described in Example 96 to give N- (R) -amino-

1-carboxypentyl] -L-lysyl-L-proline. After chromatography, the desired isomer is isolated. The mass spectrum shows the molecular ion for the disilylated species at 516 m / e, the M * - 15 ion at 501 m / e, and the peak at 399 m / e, corresponding to the loss of silylated carboxyl.

Example 99 N- _O (- / 5-amino-1'-benzyloxycarbonylpentyl) L-lysyl-L-proline.

Benzyl 2-oxo-6-phthalimidohexanoate can be treated in the manner described in Example 96 except that the debenzylation with hydrogen on palladium is omitted. From the product mixture, the desired N- ( _R ) -benzyloxycarbonylpentyl) -L-lysyl-L proline can be isolated.

Example 100 A. N - N - (1 N -carboxy-2-phenoxyethyl) -L-lysylL-proline.

Phenoxypyruvic acid (0.9 g) was dissolved in water, the pH was adjusted to 7 with dilute sodium hydroxide solution and the solution was freeze-dried. The rest

- 73 The residue was dissolved in 10 ml of ethanol and treated with? -3 ° butoxycarbonyl-L-lysyl-L-proline (0.36 g) and a molecular sieve powder ^? 4A (3.0 g). Sodium cyanoborohydride (0.18 g in 3.5 ml ethanol) was added portionwise to the mixture and the reaction mixture was stirred at room temperature until the reaction was complete. The product was isolated by absorption on a highly acidic ion exchange resin and eluting with a 2X-ene solution of pyridine in water, followed by freeze-drying to give 0.25 g of unprotected product, N - (1) Carboxy-2-phenoxyethyl) - B lysyl-L -proline. The MRI and product mass spectra are consistent with its structure.

B. N _s / - / 1'-ethoxycarbonyl-2-phenoxyethyl / -L-lysyl Lprolin.

Ethyl phenoxypyruvate was treated with N -3-butoxycarbonylL-lysyl-L-proline in the manner described in Example 100A to give N- [1- (3-hydroxycarbonyl-2-phenoxyethyl) -L-lysyl-Lproline]. The NMR spectrum showed phenyl absorption centered at 7.3 ° and broad peaks at 4.6, 4.4, 4.2, 3.7, 3.2 and 1.9 (?.

Example 101 tons A. 1CH- _with / - / 1/5 / The -karbO1 {SI-2-phenylthioethyl / -L lysyl-L-proline.

Feniltiopirogrozdena acid was treated with £ -3 ° -butoxycarbonyl-L-lysyl-L · proline as described in Example 100 a way to obtain N _with £ - / 1'-carboxy-2-phenylthioethyl / -B lysyl-L-proline . The mass spectrum shows a silylated molecular ion at 567 m / e. Chromatography afforded the desired isomer.

B. L- _O (- (1-ethoxycarbonyl-2-phenylthioethyl) N-lysyl-L proline.

Work up analogously to ethyl phenylthiopyran with 4-3 ° - 74 butoxycarbonyl-L-lysyl-L-proline in the manner described in Example 100A to give N, N - (1-ethoxycarbonyl-2- $ enylthioethyl / L-lysyl-L · - proline.

Example 102 N _s / - / 1-carboxy-2- / 57-amino-3-phenylpropyl / £>, L-lysyl-L · proline.

Ethyl 2-amino-4-phenyl-3-phthalimidobutanoate can be condensed with 2-oxo-6-phthalimidohexanoic acid, obtained by alkylation of benzyl 1,3-dithian-2-carboxylate with phthalimidobutyl bromide, followed by oxidation and oxidation. of sodium cyanoborohydride by the method described in Example 13. The resultant intermediate can then be reacted with L -proline benzyl ester hydrochloride using diphenylphosphoryl azide as described in the same example to give a mixture of M- isomers. N- (1-ethoxycarbonyl-2/5) -phthalimido-3-f phenylpropyl / -Y1- £ -ftaloil-D, L-lysyl-L · proline benzyl ester, purified by column chromatography. The product can then be treated with two equivalents of sodium hydroxide in ethanol-water for four hours at room temperature, neutralized to pH 4 with hydrochloric acid concentrate, the ethanol distilled off in vacuo, the product extracted with ethyl acetate and remove the solvent in vacuo. The resulting residue was then refluxed in ethanol containing 2 equivalents of hydradine for an hour and a half, and isolated in the manner described in Example 74 so that the desired compound was obtained.

Example 103 N- [1- (1-carboxy-2- (S) -benzoylamino-3-phenylpropyl) -P, L-lysyl-L · -proline.

The ethyl 2-amino-3-benzoylamino-4-phenylbutanoate obtained in Example 77 was nondensed with 2-oxo-6-phthalimido hexanoene

- 75 acids in the presence of sodium cyanoborohydride by the method described in Example 13. The resultant b1 - [- (1-carbethoxy-2/5) -benzoylamino-3-phenylpropyl] -N1- [beta] -phtaloyl-D, L - the lysine can then be attached to the L-proline benzyl ester hydrochloride using diphenylphosphorylazide as described in the example procedure to obtain a mixture of Ν -, (- / 1-καρ6βτοκοπ2 (6) -benzoylamino-3-phenylpropyl - N-phthaloyl-D, L-lysyl-proline benzyl ester, purified by chromatography. Treatment of this product with two equivalents of sodium hydroxide in a mixture of ethanol-water for four hours at room temperature, and after developing the mixture in the manner described in Example 102, gave 1M- _c / - (1carbons-2- (5)). -benzoylamino-3-phenyl-propyl-N-N-phthaloyl-D, L-lysyl-L-proline. By boiling this product under reflux for an hour and a half in ethanol and the presence of one equivalent of hydrazine and isolating the product as described in Example 74, the desired compound can be obtained as a mixture of isomers.

Example 104. N- _</ - / 2-amino-1-carboxy-4-methylphenyl / -D, L-lysyl-L-proline.

Trans-3-amino-4- (2-methylpropyl) -2-azetidinone, obtained in Example 78, can be reductively attached to ethanol in ethanol

2-Oxo-6-phthalimidohexanoate using sodium cyanoborohydride and molecular sieves. The product N - [- (4- (2methylpropyl) -2-oxo-3-azetidinyl) -N-N-phthaloyl-B, L-lysinbenzyl ester can then be debenzylated by hydrogenation on palladium, the free acid obtained and the benzyl ester of proline can be reacted in the presence of diphenylphosphoryl azide. The resulting product can then be debenzylated as described above to give N - [(4-2-methylpropyl) -2-oxo-3-azetidinyl] -N-N-phthalyl-D, L lysyl-L -proline. The phthaloyl group may be removed in ethanol by a solution of one molar equivalent of hydrazine at room temperature to give b1- _a (- (4/2-methylropyl) -2-oxo-3azetidinyl) -P, L-lysyl-L-proline. By hydrolysis with a dilute solution of sodium hydroxide, dissolution of the β-lactam ring can yield N- (- (2-amino-and-carboxy-4-methylpentyl) -P, L-lysyl-L-proline.

Example 105. N - N - N - benzamido-! -carboxy-4-methylpentyl / D, L-lysyl-L-proline. N-cis- [4- (2-methylpropyl) -2-oxo-3-azetidinyl] -H- ⁴ S -3-butoxycarbonyl-O, L-lysine benzyl ester can be obtained from 3-trans-4- (2-methylpropyl) -2-azetidinone (Example 78) and benzyl 3 ° -butoxy carbonylomino-2-oxohexanoate. The benzyl group can be cleaved by hydrogenation and the resulting product is coupled with the L-proline benzyl ester, the resulting product N- ₍ N- (4- (2-methylpropyl) -2-oxo-3-azetidinyl) -1H- The 3 ° -butoxycarbonyl-P, L-lysyl-L-proline benzyl ester can then be debenzylated by hydrogenation and the L-lactam hydrolyzed with a dilute base to give N - / - (2-amino-1 · carbonyl- 4-methylpentyl) -N- -3-butoxycarbonyl-D, L-lysyl) -L-proline. After benzoylation with benzoyl chloride in an organic solvent and cleavage of tert. With a trifluoroacetic acid protecting group, N-O (- (2-benzoamido-L-carboxy-4-menthylpentyl) -1), L-lysylL-proline can be obtained.

Example 106. L1 - [- (1- (5) -carbony-3-p-chlorophenylpropyl) -lysyl-trans-4-methoxy-L-proline.

Methyl-trans-4-methoxy-L-proline hydrochloride is attached to N - ^ - Tepq.Boc-N-t-Cbz-L-lysine using

- 77 Dicyclohexylcarbodiimide and triethylamine in methylene chloride as described in Example 20. The resulting product was purified by chromatography, the ester was hydrolyzed, and the tert.-Boc-protecting group was removed as described in the above example. This t-Cbz-L-lysyl-L-4-methoxyproline is then reductively coupled to 2-oxo-4-p-chloro-enyl butyric acid, obtained from p-hydrochloric acid, ethyl ester, by alkali-catalyzed condensation with diethyloxalate, followed by decarboxylation with anhydrous hydrochloride in acetic acid, in the presence of sodium cyanoborohydride, and further treating the mixture in the manner described in Example 24 to give a mixture of isomers of N- ₍ N) -carboxy-3- p-chlorophenylpropyl) -N-Cbs-L-lysyl-trans-4-methoxy-b proline. The benzyloxycarbonyl protecting group is cleaved by catalytic hydrogenation on palladium on carbon as a catalyst in a known manner.

After chromatography the resulting mixture, the desired isomer is isolated. The mass spectrum has peaks at 584 (M ^{+ 1} ), 568 (m ⁺ -15), 466, 397, 366 and 352 (base peak) for the dissolved material.

Example 107. N - [(N) -carboxy-3-p-chlorophenylpropyl] -L lysyl-L -4-thiazolidine carboxylic acid.

N- <Z-Tepq.Boc-bI-is-Cl ^ -L-lysine can be attached to I-thiazolidine-4-carboxylic acid, benzyl ester, hydrochloride, the product obtained is purified by chromatography, the ester is hydrolyzed and tert.Remove the Boc protecting group as described in Example 20. By reductively coupling the resulting product with 2-oxo-4-p-chlorophenylbutyric acid, a mixture of the N - / - (N-carboxy-3-chlorophenylpropyl) - 78 N-N-Cb2 isomers can be obtained in the manner described in the example. -L-lysyl-L · -4-thiazolidine carboxylic acid.

Removal of the benzyloxycarbonyl protecting group by treatment with hydrobromic acid in acetic acid at room temperature in a standard manner for peptide chemistry, distilling the solvent in vacuo, pouring the residue with water and freeze-drying can give the desired product.

Example 108. N - N - (1-carboxy-3-p-chlorophenylpropyl) b lysyl-D, L-trans-5-methylthiazolidine-4-carboxylic acid.

3.7 from trans-5-methylthiosolidine-4-carboxylic acid ethyl ester, hydrochloride derived from N, -bromo crotonic acid and thioacetamide, acid hydrolysis of the resulting product to L methylcysteine and subsequent reaction with formaldehyde analogous to the method used by N, F. Nutt et al., Abstractsj of the Btn American Peptide Symposium, Washington, dC

(1979), 1 + 16, P.95). was reacted with 7.4 r N - / - tert.-Boc-C6-L-lysine using 2.8 ml of triethylamine and

4.5 g of dicyclohexylcarbodiimide in methylene chloride as described in Example 20. By reductively coupling the resulting intermediate to 2-oxo-4-p-chlorophenylbutyric acid using sodium cyanoborohydride and then removing the benzyloxycarbonyl protecting group in the manner described in Example 107, the H - / - / I · - can be obtained carboxy-3-chlorophenylpropyl) -1H-lysyl-D, L-trans-5-methylthiazolidine-

4-carboxylic acid in the form of a mixture of isomers.

Example 109. N - [(N) -carboxy-3-p-chlorophenylpropyl] -L lysyl-L · -3,4-dehydroproline.

Condensation of L -3,4-dehydroproline, ethyl ester, hydrochloride with L1 - / - tert.-Boc-L1- [beta] -Cr-11-lysine, removal of tert.-Boc protective group with 4 M hydrogen chloride in ethyl acetate,

- 79 reductive coupling of the intermediate with 2-oxo-4-chlorophenylbutyric acid. Removal of the protecting group with hydrobromic acid in acetic acid and further treatment of the reaction mixture in the manner described in Example 107 gave a mixture of the isomers of N, N -b-carboxy-3-p-chlorophenylpropyl) -L-lysyl-L -3,4-dehydrolrolin.

Example 110. 1'1- (1-Carbonsi-4-methylphenyl) -L-alanylL-proline.

A solution of 5-methyl-2-oxohexanoic acid (1.44 g) and L-alanyl-L-propane (0.37 g) in five ml of water at pH 7 was treated with sodium cyanoborohydride (0.31 g). After stirring at room temperature for five days, the reaction product was absorbed onto a highly acidic ion exchange resin and eluted with a 2 ° 4-pyridine solution in water to give 0.6 g of freeze-dried solid. Part of it (0.2 g) was purified by chromatography on LH-20 to give 0.18 g of N- (1-carboxy-4-methylphenyl) -L-alanyl-L-proline. The NMR and mass spectra are in accordance with the expected structure. Diastereomers can be isolated by chromatography. In the mass spectrum, the pins of the disilylated material were observed at 458 (M ⁺ ), 443 (M ⁺ -15), 341, zO1 and 244 m (e (base peak)).

Example III-N- (1R) -Ethoxycarbonyl-4-methylphenyl) -L alanyl-L-proline.

Ethyl 5-methyl-2-oxohexanoate (3.44 g) and L-alanyl-L proline (0.74 g) were stirred in 15 ml ethanol with 6 g powder molecular sieve 4A. Sodium ions borohydride solution (0.23 g) was added dropwise over several days.

- 80 in ethanol. The ethanol was then removed in vacuo, the product was absorbed on a highly acidic ion exchange resin and eluted with a 2% solution of pyridine 8 in water to give 1.08 g of 1-l- (1-ethanesinarbonyl-4-methylphenyl) -L-alanyl- L -proline. Part of the product was purified on column L H-20 (chromatographic) for spectral analysis. The NMR spectrum is consistent with the structure. The mass spectrum showed a peak at 414 (silylated molecular ion - 15). Chromatography of the product gave the desired isomer.

Example 112. N- (L-anarbonyl-3-p-phenoxyphenylpropyl) -L alanyl-L-proline.

A mixture of 2-oxo-4-p-phenoxyphenyl butyric acid obtained by reacting p-phenoxyphenyl Grignard reagent with ethylene oxide, converting the resulting alcohol into bromide and condensing the product with ethyl 1,3-dithian-2-carboxylate. Following oxidative cleavage of dithianium followed by alkaline hydrolysis, keto acid is obtained and L-alanyl-L -proline in water at pH adjusted to 7 with a diluted base is treated with excess sodium cyanoborohydride. The product N- (Ircarboxy-3-p-phenoxyphenylpropyl) -L-alanyl-L-prodrug was isolated by chromatography. The mass spectrum of the disilylated material contains pins at 584 (M *), 569 (M ⁺ -15), 468 and 370 m (e) (base peak).

Example 113. N- (1-Ethoxycarbonyl-3-p-phenoxyphenylpropyl) -L alanyl-L-proline.

Ethyl 2-oxo-4-phenoxyphenylbutyric acid, prepared in the manner described in Example 112, with only the final alkaline hydrolysis being omitted, is condensed with L-alanyl-L-proline in the presence of sodium cyanoborohydride to give

- 8] N- (1-ethoxycarbonyl) -3-p-phenoxyphenylpropyl] -L-alanyl-L · proline.

Example 114. L1- [L-carboxy-3-phenylpropyl] -L-alanyl-L D, L -3,3-dimethyl-proline.

3,3-Dimethyl-N, L-proline is prepared from 3-methyl-2-butenal by the method of Cox, J. Snem, Soc., 1964, 5024 ^and converted to methyl ester hydrochloride with methanolic acid. The resulting product was attached to tert.-Boc-alanine, then the intermediate was condensed with 2-oxo-4-phenylbutyric acid by the method described in Example 47 to give a mixture of isomers of the desired product.

Example 115.

A. L- (1-carboxy-3-phenylpropyl) -N-5-benzyl-cysteinyl L-proline.

Condensation of 1-M-tert. Boc-5-benzylcysteine with L-proline tert. butyl ester in the presence of nadicycloxyl carbodiimide in the usual way results in the production of N - (N-tert). Boc-benzylcysteine (N) -b-proline-tert. Boc butyl ester. This blocked dipeptide was then treated with 4N hydrogen chloride in ethyl acetate at 0 ° C to give L - (N - benzylcysteinyl) - E -proline. After treatment of this dipeptide with 2-oxo-4-phenylbutyric acid in the presence of sodium cyanoborohydride, N- (1-carboxy-3-phenylpropyl) -L (5-benzylcysteinyl) -L-proline is formed in the form of a mixture of isomers.

B. 1- (1-carboxy-3-phenylpropyl) -L-cysteinyl-L-proline. The N- (1-carboxy-3-phenylpropyl) -L-5-benzylcysteinyl-b proline prepared above in part A is treated with sodium in liquid ammonia to give the desired compound.

82 Example 116. N - N - (1-carboxy-3-phenylpropyl) -1 H-ornyl-L proline.

N-S-Tepq.Boc-L-ornyl-L-proline and 2-oxo-4-phenylbutyric acid were condensed in the presence of sodium cyanoborohydride as described in Example 54. The protecting group is removed from the product obtained using ethyl acetate with 4 N hydrogen chloride gas. The crude diastereomer hydrochloride was absorbed onto a highly acidic ion exchange resin and eluted with 2X aqueous pyridine solution. The mass spectrum shows a molecular ion at 355 m / e for the product minus 36. The NMR spectrum corresponds to the structure of the product

Example 117. Y1- _with / - / 1/5 / The -etoksikorbonil-3-phenylpropyl / -L lysyl-L-proline.

Ethyl 2-oxo-4-phenylbutyrate (2.58 g) and N-tert.-Boc-L lysyl-L-proline (859 mg) were dissolved in absolute ethanol (5 µm) and (2.0 g) milled molecular sieves were added to the solution. 4A. At the completion of the reaction, the sieve is removed by filtration. After evaporation of the filtrate, the residue was dissolved in water, extracted with ester and absorbed on a highly acidic ion exchange resin. Elution with a 2 ^ solution of pyridine in water afforded 639 mg of the crude product ethoxycarbonyl-3-phenylpropyl) -L-1-tert-Boc-L-proline. The protecting group is removed with ethyl acetate which is 4 N in hydrogen chloride gas. The resulting hydrochloride was absorbed onto a highly acidic ion exchange resin and eluted with 2L aqueous pyridine to give 270 mg of product. The mass spectrum shows a molecular ion at 678 m / e for the disilyliron species plus one. The NMR spectrum is consistent with the structure.

- 83 Chromatography afforded the desired isomer.

Example 118. N-7- (1-Carboxy-3-phenylpropyl) -N-N-N-dimethyl-L-lysyl-L-proline.

The B1 - / - tert.-Boc-L1- [beta] -Cb2-L-lysyl-L-proline benzyl ester was reductively methylated in formaldehyde / 10% palladium / charcoal at 40 pounds / square inch of hydrogen pressure. / - tert.The protecting group is cleaved with ethyl acetate, which is 4N in hydrogen chloride gas. In the method described in Example 54, 2-oxo-4-phenylbutyric acid and N - N - N - dimethyl-L-lysyl-L-proline were condensed in the presence of sodium cyanoborohydride. The mass spectrum shows a molecular ion at 415 for the product, minus 18. The NMR spectrum is consistent with the structure.

EXAMPLE 119 N _x / - / 1/5 / The carboxy-3-phenylpropyl / -L -lizilL proline.

N- [1- (1-carboxy-3-phenylpropyl) -L-lysyl-L-proline, a mixture of diastereomers obtained by the method of Example 57B, was purified by gel filtration on a L H-20 chromatographic column. The XAD-2 column obtained by the method described in Example 25 was equilibrated at 53 ^P C with 0.1 M-eunion hydroxide containing 4% acetonitrile. The isomer mixture (250 mg) was dissolved in 10 ml of the solvent mentioned above and added to the column. When the column is eluted with the same solvent, the first isomer appears at an elution volume in the range of 320-360 ml. The second isomer appears in the range of 450-540 ml of eluate. The intermediate fractions contain a mixture of isomers. the fractions containing the first isomer are freeze-dried to give 72 mg of a fluffy white solid, which is the more active isomer and is an SSS configuration by analogy with the more active G1- / - / 1-carbonyl isomer. 3-phenylpropyl-L-alanyl-L-proline, which was found to have an SSS configuration by X-ray analysis. In thin layer chromatography on silica gel in a 1: 1: 1: 1: 1: 1 ratio of ethyl acetate / n-butanol / water / acetic acid, the solid gave a single spot having an Rf of 0.43.300 MHZ, NMR spectrum indicated triplet for the methin proton y to the phenyl substituent at 3.40 ppm. When the fractions containing the second isomer are freeze-dried, 72 mg of a white powdery solid is obtained, which, by thin layer chromatography on silica gel, gives only a spot with an Rf of 0.39.300 'MHZ, the NMR spectrum shows triplets of the methine proton relative to the phenyl substituent at 3.61 ppm.

Example 120. N- [1- (1-Carboxy-3-Phenylpropyl) -L-N-octylL-lysyl-L-proline.

According to the method described in Example 54, 2-oxo-4-phenylacetic acid and N- [beta] -acetyl-L-lysyl-L-proline were condensed in the presence of sodium cyanoborohydride to give N, N - (N) -carboxy- 3-phenylpropyl- [N] -acetyl-l-lysyl-L-proline. The NMR spectrum of the product obtained is consistent with its structure. the mass spectrum shows a molecular ion at 663 for the trisilylated species.

Example 121. N - N - (1-carboxy-3-phenylpropyl) N- arginylL-proline.

The required dipeptide is obtained by condensation of N-Tepa.Boc-N-u / -HHTpo-L-arginine and L-proline benzyl ester hydrochloride by the carbodiimide method. The protective-tert. The Boc group was removed by a known method with 4 N hydrochloric gas in ethyl acetate, and the resulting N-VU-hhtpo-L * arginyl-L-proline benzyl ester was condensed with 2-oxo-4-phenylbutyric acid as described in Example 24.

The reaction yields N - (-) carboxy-3-phenylpropyl (N-O-nitro-L-arginyl-L-proline benzyl ester) in a relatively low yield (25-33%). This compound / 159 mg / are dissolved in a mixture /2.5 ml / acetic acid / water / methanol / 84%, 8X, 8 $ / and hydrogenated at 40 pounds / inch ^-1-pressure hydrogen at room temperature in the presence of 130 mg of 10% palladium / carbon for the simultaneous removal of the W-nitro and benzyl ester protecting group. The catalyst was filtered off and the filtrate was evaporated to dryness (94 mg), the water-soluble portion of the resulting residue was freeze-dried to give a fluffy white substance (90 mg). This solid is the acetate salt of the desired product and the free base is obtained by absorption on a highly acidic ion exchange resin, washing with water and then eluting with a 2% solution of pyridine in water. Freeze-drying of the product-rich fractions afforded 60 mg of N- (N'-carboxy-3-phenylpropyl) -N-arginyl-L-proline. The NMR spectrum is consistent with the structure, the mass spectrum shows a molecular ion at 793 for the pentasilylated species.

Example 122. N- (1'Carboxy-3-phenylpropyl) -L-histidylL-proline.

In the method described in Example 54, 2-oxo-4-phenylacetic acid and L-histidyl-L-proline were condensed in the presence of sodium cyanoborohydride to give N- / L-carboxy-

3-phenylpropyl-L-histidyl-L-proline. The product was purified by gel filtration on a chromatographic column (L H-20) in methanol. Its NMR spectrum is consistent with the structure. The mass-center

86 shows a molecular ion at 657 for the disilylated species.

Example 123. N - [- (L-Carbony-2- (3-indolyl) ethyl] -L lysyl-L-proline.

In the method described in Example 54, the indole-3-pyruvic acid is condensed with N-N-Tepq.Boc-L-lysyl-L-proline in the presence of sodium cyanoborohydride. The Boc protecting group is cleaved from the product with 4 N hydrogen chloride in ethyl acetate. the resulting hydrochloride is absorbed on Dowex 50 (H ⁺ ) and eluted with a 2% solution of pyridine in water. Freeze-drying of the product-rich fractions results in the free base being a light brown fluffy substance. The NMR spectrum of this product is consistent with its structure. The mass spectrum shows a molecular ion at 718 for the trisilylated species.

Example 124. N- / N- Ethoxycarbonyl-4-methylpentyl] -Lysyl-L proline.

Dissolve 2-oxo-4-methyl-ethylpentanoate (2.75 g) in 150 ml ethanol containing 16 g powder molecular sieve 4A. Hydrogenates at an absolute pressure of 40 pounds / inch of hydrogen at room temperature over 1 g of 10% palladium / carbon. After 1 mole of hydrogen is taken up, the catalyst is filtered off, the filter cake being carefully washed with ethanol. The solvent was evaporated to give 5.87 g of an oil. The oil was suspended in water, the pH of the medium was adjusted to 8.5, and extracted with ethyl acetate (3 x 60 ml) to remove neutral materials. The pH of the medium was adjusted to 7, saturated with sodium chloride and extracted with ethyl acetate (3 x 100 ml). The product solution was dried over anhydrous magnesium sulfate. The ethyl acetate was evaporated to give 4.38 g of crude N - [- (L-ethoxycarbonyl-4-methylpentyl) -L-N-tert-Boc-L-lysyl-L-proline. The tert.-Boc-protecting group was removed as usual with 4 N hydrogen chloride gas in ethyl acetate. Conversion of the resulting hydrochloride to a free base with a highly acidic ion exchange resin, eluting with a 2% solution of pyridine in water. Freeze-drying of the product-rich fractions gave 2.1 g of hygroscopically friable material. The NMR spectrum of this product is consistent with the structure of N - [- (N-carbethoxy-4-methylphenyl) -L-lysyl] -proline. The mass spectrum peaks at 472 for the monosilylated molecular ion plus unity.

Example 125. L1 - [- (1-carboxy-4-methylpentyl) -L-lysyl-L proline.

N1- [1- (1-Ethoxycarbonyl-4-methylpentyl) -L-lysyl-L-proline is hydrolyzed to the corresponding carboxylic acid with stirring in aqueous sodium hydroxide solution (2.5 equivalents) at room temperature for several days. The reaction mixture was acidified to pH 5, absorbed on a highly acidic ion exchange resin, and eluted with a 2% solution of pyridine in water. The product rich fractions were freeze-dried to give N- / L-carboxy-

4-methylpentyl N-L-lysyl-L-proline as a white fluffy substance. The NMR spectrum is consistent with its structure. The mass spectrum shows a molecular ion at 516 for the disilylated species.

Example 126. N - [- (I-carboxy-3-phenylpropyl) -L-leucyl-trintophan.

According to the method described in Example 54, 2ooxo-4-phenylbutyric

- 88 Acid and L-leucyl-L-tryptophan are condensed in the presence of sodium cyanoborohydride. The product was freeze-dried in a dioxane / water mixture until it was only slightly water soluble. The NMR spectrum is consistent with the structure. The mass spectrum shows a molecular ion at 695 for the trisilylated species.

Example 127.

A. Tablets. The typical tablet contains N- (1- (5) -ethoxycarbonyl-3-phenylpropyl) -L-alanyl-L-proline (25 mg), pre-gelatinized starch (SP (82 mg), microcrystalline cellulose (82 mg) and magnesium stearate (I mg). Similarly, for example, N- (1- (5) -Narboxy-3-phenylpropyl) -L-lysyl-L-proline (20 mg) can be labeled in place of N- (1- (R) ethoxycarbonyl-3-phenylpropyl). N-alanyl-L-proline, with the composition of pre-gelatinized starch, microcrystalline cellulose and magnesium stearate described above.

B. Combination tablets. A combined diuretic tablet, such as hydrochlorothiazide, typically contains b1- (1/6-ethoxycarbonyl-3-phenyl-propyl) -1 galanyl-L-proline (7.5 mg), hydrochlorothiazide (5mg), pre-gelatinized starch // 5P mg), microcrystalline cellulose (82 mg) and magnesium stearate (I mg). Tablets with, for example, N- (1/5 ') -carboxy-3-phenylpropyl (L-lysyl-L -proline) (5 mg) and hydrochlorothiazide (50 mg) are prepared using said active ingredient in place of b1- (1) 5-Ethoxycarbonyl-3-phenylpropyl. in the composition described above.

B. Injections. The typical injectable form contains N- (I) S (carboxy-3-phenylpropyl) -L-alanyl-L-proline (5.42 mg), anhydrous sodium phosphate (11.4 mg), benzyl alcohol (0.01 ml) and water for injections). 1.0 ml /. Similarly, this formulation can be prepared by using, for example, N- (I) 5 (carboxy-3-phenyl-propyl) -L-lysyl-L-proline in place of N- (1/5) -ethoxycarbonyl-3 -phenylpropyl / -L-alanyl-L -proline. Such an injection form may also contain a pharmaceutically effective amount of another antihypertensive or diuretic agent, for example hydrochlorothiazide, as described in Example B above.

G. Suppositories. Typical rectal administration suppositories contain N- (1/5) -ethoxycarbonyl-3-phenylpropyl) -L alanyl-L · -proline (1-25 mg) butylated hydroxyanisole (0.08-1.0 mg), disodium calcium ethylenediaminetetraacetate / 0.25-0.5 mg) and polyethylene glycol (775-1600 mg). Other suppository forms may be prepared, for example, by replacing the above-mentioned active substance with N- (1- (S) -carboxy-3-phenylpropyl) -L-lysyl-L-proline and, for example, by substituting disodium calcium ethylenediaminotetraacetate with butylated hydroxytoluene (0.04-0.08 mg) and polyethylene glycol with hydrogenated vegetable oil (675-1400 mg), such as Suppository L, Century P5, Century

I. ', Vitepsoli and others. similar. Further, these suppository forms may include other antihypertensive or diuretic agents in a pharmaceutically effective amount, as described, for example, in Parts B and B of the same example.

Example 128 N- (1-ethoxycarbonyl-3- (3-indolylpropyl) -L alanyl-L-proline.

A solution of L-alanyl-L-proline (0.35 g) and ethyl-4 (3-indolyl) -2-oxobutyrate (1.0 g) in 30 ml of ethanol is hydrogenated with 10% palladium / charcoal in the presence of 3.2 g powder molecular sieve (34A) at an absolute hydrogen pressure of 40 pounds per square inch. After the reaction is complete,

- 90 the suspension is filtered and the filtrate is concentrated to dryness. The ottanol oil was dissolved in 5 ml of methanol and chromatographed on a Sephadex L H-20 column to give 0.67 g of crude product as an orange oil. A portion of the crude product (0.3 g) was purified by dissolving in water and extracting with ether. The product was recovered from the aqueous phase by diluting with an equal volume of saturated sodium chloride solution and extracting with ethyl acetate to give 80 mg of N- / I-ethoxycarbonyl-

3- (3-indolyl) propyl] -L-alanyl-L -proline as a yellow oil. The mass spectrum shows peaks at 559 for the disilylated product and at 632 for the tri-silylated product.

Example 129. Benzyl N- [1- (6) -carbonsi-3-phenylpropyl] -1'-alanyl-L · -prolineate.

Thionyl chloride (13.1 ml) was added to 150 ml of benzyl alcohol while maintaining the temperature below 0 ° C (exothermic reaction). 15 g of N- (1/4) -carboxy-3-phenylpropyl) -L-alanyl-L · proline were added portionwise to the cold solution. The cooling bath was removed and the reaction mixture was stirred at room temperature overnight. After heating to 45 ° C in vacuo to remove the dissolved gases, the reaction mixture was diluted with 500 ml of ether and washed with 10 x 100 ml of water. The solid which appears in the organic layer was filtered and dried to give 4.9 g of crude monobenzyl ester.

The combined aqueous extracts were neutralized with sodium bicarbonate, yielding <2 a second crop of 1-4 g of product. The product from both crops was collected and part of it (1.0 g) was recrystallized from ethanol-water to give 0.95 g of pure monobenzyl ester, m.p. 120-125 ° C. The spectrum

91 indicates that the benzyl ester is attached to the proline ring. Example 130. Ethyl N- (1- (S) -ethoxycarbonyl-3-phenylpropyl) -L alanyl-L-proline hydrochloride.

A solution of 3.0 g of N- (1/5) -ethoxycarbonyl- (3-phenylpropyl) L-alanyl-L-proline in 50 ml absolute ethanol was saturated with hydrogen chloride gas at 0 ° C. After standing at 0 ° C overnight, the hydrogen chloride and ethanol were removed in vacuo to give 3.1 g of an oil, which, when freeze-dried, gave a white solid. Thin layer chromatography and high performance liquid chromatography (HPLC) indicate that the product consists of only one component. The NMR spectrum lists two ethyl groups for the aromatic nucleus. The mass spectrum shows a molecular ion at 404 m / e.

Example 131. N 1- (1-Ethoxycarbonyl-3- (2-chlorophenyl) propyl) L-alanyl-L-proline.

In the method described in Example 26, ethyl-2-oxo-4- (2-chlorophenyl) butyrate and L-alanyl-L-proline were condensed in the presence of sodium cyanoborohydride to give N- (ethoxy) carbonyl-3- (2-). chlorophenyl (propyl) -L-alanyl-L-proline. The mass spectrum showed a peak at 411 (M + 1) and a strong peak at 392 (M - 18).

Example 132. b1- (1-ethoxycarbonyl-3- (4-fluorophenyl) propyl) L-alanyl-L-proline.

In the method described in Example 26, ethyl 2-oxo-4- (4-fluorophenyl) butyrate and L-alanyl-L-proline were nondensed in the presence of sodium cyanoborohydride to give N (1'-ethoxycarbonyl) -3- (4-fluorophenyl) -propyl] -L-alanyl-L-proline. The mass spectrum shows a molecular ion at 394 and strong peaks at 376 (M-18, water) and 375 (M-19, fluorine). The NMR spectrum is consistent with the structure.

- 92 The elemental analysis indicates that the product is in the form of sesquihydrate.

Example 133. N- (T-Ethoxycarbonyl-3- (2-naphthyl) propyl) -L alanyl-L-proline.

by the method described in Example 26, ethyl 2-oxo-4- (2-naphthyl) butyrate and L-alanyl-L-proline were condensed in the presence of sodium cyanoborohydride to give N- (1-heteroxycarbonyl) - (3-) 2- naphthyl (propyl) -L-alanyl-L-proline. The mass spectrum shows a molecular ion at 426 and a peak at 408 (M-18, water). The NMR spectrum is consistent with the proposed structure. Elemental analysis indicates that the product is in the form of a hemihydrate.

Example 134. N-Z- (N-carboxy-3- (2-chlorophenyl) propyl) -Lysyl-L-proline.

In the method described in Example 26, ethyl 2-oxo-4- (2-chlorophenyl) butyrate and N-tert. Boc-L-lysyl-L-proline were condensed in the presence of sodium cyanoborohydride to give NZ- (I - ethoxycarbonyl-3- (2-chlorophenylpropyl) -N-N-tert.-Boc L-lysyl-L-proline.

The tert.-Boc-protecting group is removed by treatment with

N hydrogen chloride gas in ethyl acetate and then the ethyl ester group is salted with a dilute solution of sodium hydroxide to give N - / - (1-carboxy-3- (2-chlorophenyl) propyl / L-lysyl-L-proline after absorption on a strong acid ion exchange resin and elution therefrom. Elemental analysis shows solvation with a molecule and half water. The NMR spectrum is consistent with the proposed structure of the compound. The mass spectrum of the silylated sample shows a peak at 568, which corresponds to a mass loss of 15 (methyl) disilylated species.

93 Example 135. N-7- (L-carboxy-3- (4-fluorophenyl) propyl) -

L-lysyl-L-proline.

In the method described in Example 134, ethyl 2-oxo-4- (4-fluorophenyl) butyrate and N, N-tert-Boc-L-lysyl-L-proline were condensed in the presence of sodium cyanoborohydride. The resulting product was released in the manner indicated in Example 4 to give N-, N- (1-carbony-3- (4-fluorophenyl) propyl) -L-lysylL-proline. Elemental analysis indicates that the product is in the form of sesquihydrate. The NMR spectrum confirms the proposed structure. The silylated sample mass spectrum shows a molecular ion at 711 for the tetrasilylated species.

Example 136. 1M- ₃ / - / b-carboxy-3- / 2-naphthyl / propyl / -L lysyl-L-proline.

By the method described in Example 134, replacing ethyl 2-oxo-

4/2-Chlorophenyl / butyrate with ethyl 2-oxo-4- / 2-naphthyl / butyrate to give N- ₃ / - / 1-carboxy-3- / 2-naphthyl / propyl / ^ lysyl-L proline. The NMR spectrum is in accordance with the proposed structure, the mass spectrum shows a molecular ion at 599 for the disilylated species.

Example 137. Ethyl 2-oxo-4- (2-thianyl) -butyrate.

A mixture of phosphorus tribromide (18.95 g) and pyridine (3.76 cm ³ ) in 10 ml of benzene was cooled to a temperature of 5 ° C and treated with a solution of 2- (2-thianyl) -ethanol (25 g) in 10 ml of benzene containing 1.25 cm ^{3 of} pyridine at such a rate that the reaction temperature never exceeds 0 ° C. The mixture was then allowed to warm to room temperature, stirring overnight. The reaction mixture was then concentrated on a water pump at 30-40 ° C to remove solvent and other volatiles, and the residue was subjected to

- 94 of Claisen distillation under vacuum provided by a water pump to obtain 21 g of crude product / bw. 115-119 ° C.

This product was then subjected to fractional distillation in 0.3 mm mercury to give 19 g of pure 2- (2-cyanyl) ethyl bromide / mp. 50 - 51 ° C; purity 99.9%, determined by gas chromatography /. This bromide was dissolved in dry tetrahydrofuran (40 ml) and added dropwise with stirring to a mixture of magnesium chips (2.4 g) and 15 ml of dry tetrahydrofuran in a nitrogen atmosphere. The mixture was then stirred and refluxed for one hour, cooled to room temperature and added dropwise under a nitrogen atmosphere and stirred to a mixture of diethyl oxalate (17.5 g) in 25 ml of dry THF at -15 ° C on the bathroom. After completion of the reaction, the mixture was stirred at -10 ° C for 15 min, then allowed to warm to 10 ° C and treated with 60 ml of saturated ammonium chloride solution. The aqueous layer was extracted twice with toluene, the combined organic layers were dried over magnesium sulfate and concentrated in vacuo to give 16.7 g of a mixture containing in% by weight. by gas chromatographic analysis: solvent 26; diethyl oxalate 10.3 and ethyl 2-oxo- (2-thienyl) butyrate 63.4. The NMR spectrum in deuterated chloroform is consistent with this mixture, showing the presence of two specific types of ethyl esters, and the mass spectrum shows a peak at m / e = 212 for the molecular ion of one keto-ester together with the fragmentation model corresponding to the structure.

Example 138. N- (L-Ethoxycarbonyl-3- (2-thienyl) propyl) L-alanyl-L-proline.

Add 4 g of molecular sieve type 4A (powder) to

- 95 mixture of L-alanyl-L-proline (0.93 g) and ethyl-2-oxo-4- (2-thienyl) butyrate (3.18 g) in 25 ml of anhydrous ethyl alcohol. The mixture was stirred for 15 minutes at room temperature followed by the slow addition of sodium cyanoborohydride solution (0.785 g) in 15 ml of anhydrous ethyl alcohol over three and a half hours and stirred overnight at room temperature. The reaction mixture was slightly acidic with glacial acetic acid, filtered, and the filtrate was concentrated in vacuo to give an orange oil.

Extraction of the chloroform solution of this oil with 10% potassium bicarbonate solution, followed by acidification (to pH 3) bicarbonate extracts with chloroform and after drying and evaporating the solvent, 400 mg of the desired substituted dipeptide is obtained. The NMR spectrum in deuterated chloroform shows the thienyl proton as a multiplet at 7.0 ppm, two active protons at 6.2 ppm as a wide singlet, six methyl protons as a wide multiplet at 1.22-1.43 ppm and a series of multiplets in the range of 1.8-4.4 ppm.

Example 139. N- (1-carboxy-3- (2-thienyl) -propyl) -L-lysyl-L-proline.

Add 6.95 g of the molecular sieves of type 4A (powdered) to a mixture of L-N-N-Boc-lysyl-L-proline (2.98 g) and ethyl-2-oxo-4- (2-thienyl) -butyrate). 6.4 g / in 40 ml of anhydrous ethyl alcohol. The mixture was stirred at room temperature for 45 min, followed by slow addition (for four and a half hours) of sodium cyanoborohydride solution (1.3 g) in 25 ml of anhydrous ethyl alcohol, stirring overnight at room temperature, which the reaction mixture was made slightly acidic using ice

- 96 acetic acid, filtered and concentrated in vacuo to give an orange foam (16.4 g). Rubbing this crude product with three portions of ether and decanting the ether solution removes a significant amount of excess ketoester. The ether-insoluble oil was stirred in 150 ml of 4 N hydrogen chloride / ethyl acetate at 0 ° C for one hour and poured into 1 liter of cold ether, filtered and concentrated in vacuo to give an orange oil. A solution of this unblocked product in 250 ml of ethyl alcohol is treated with 5 g of sodium hydroxide in 35 ml of water and stirred at room temperature overnight. Subsequent acidification with 0.5 N hydrochloric acid, concentration in vacuo and subsequent purification by elution from a 150 ml Dowex 50W-X2 (H +) column with water containing 2% pyridine yielded the substituted dipeptide, such as dicarboxylic acid. The NMR spectrum in deuterated water D CI is consistent with the structure.

Example 140 N- (1-carboxy-3-phenylpropyl) -L-olanyl-Hamoisobutyric acid.

A mixture of 3.9 g of methyl 7-amino-isobutyrate hydrochloride, 8.7 ml of triethylamine and 11.9 g of Boc-L-alanine in 135 ml of methylenedichloride was prepared. A solution of 14.2 g of dicyclohexylcarbodiimide in 20 ml of methylene chloride was added and the mixture was stirred at room temperature for 48 hours. The mixture was filtered and the filtrate concentrated, the residue partitioned between water and ethyl acetate. The organic phase was washed with 10% citric acid, with water, with 5% sodium bicarbonate solution and with water. It was dried over magnesium sulfate and concentrated to give the crude dipeptide. The crude product was purified * 97 on silica gel eluting with a 7: 3 mixture of hexane-ethyl acetate. 1.2 g of this dipeptide were dissolved in 30 ml of 4N hydrogen chloride / ethyl acetate and held at room temperature for one hour. The mixture was concentrated and dried by freezing from water to give L-alanyl-7-aminoisobutyric acid, methyl ester, hydrochloride.

A solution of 688 mg of this amino acid is prepared as an ester of 949 mg of ethyl 2-oxo-4-phenylbutyrate and 328 mg of sodium acetate in 10 ml of ethanol. This mixture is hydrogenated over 0.5 g of 10% palladium / carbon as catalyst and at an absolute pressure of 40 pounds / square inch of hydrogen. Filter and concentrate the filtrate. 4 ml of 7: 3 hexane / ethyl acetate mixture was added, filtered and the filtrate was chromatographed on silica gel to give methyl b1 - / - ethoxycarbonyl-

3-Phenylpropyl (N- (L) -alaniline-N-amino-isobutyrate.

Elemental analysis: Calculated for CSS ⁿ ZO ^ 2 ^ ^5: C, 63.30;

H, 8.24; N, 7.38.

Found: C ₍ 63.58; H ₍ 8.13 and N, 7.27.

The NMR spectrum is consistent with the structure.

460 mg of this diester was stirred with 3 ml of 2 N sodium hydroxide at room temperature overnight. The resulting solution was loaded onto a column of Dowex 50 (H +) and eluted with a 5% solution of pyridine in water. Concentration of the appropriate fractions gave N- (1-carboxy3-phenylpropyl) -L-alanyl-c). -Amino-substituted acid.

Elemental analysis. Calculated for C-2H27M2O52H2O2 C, 54.82; H, 7.58; N, 7.52. Found: C, 54.94; H, 7.50; and N, 7.48.

The NMR spectrum is consistent with the structure of the compound.

- 98 Example 141. N- (1-Carboxy-3-phenylpropyl) -L-alanyl-L / methylphenylalanine.

The compound was prepared according to the method described in Example 140 starting from L - N -methylphenylalanine methyl ester hydrochloride in place of -aminoisobutyric acid, methyl ester, hydrochloride.

Example 142. N- (N-Carboxy-3- (4-) N-benzylimidazolyl) propyl-L-alanine-L-proline.

The excess of 4- (4- (1H-benzylimidazolyl) -2-non-butyric acid, obtained by dibenzylation of 3- (4-imidazolyl) propionic acid and subsequent condensation with diethyl oxalate, followed by acid hydrolysis and decarboxylation, condenses with L-alanyl-L-proline in water (pH 6.7) in the presence of excess sodium cyanoborohydride. After three days at room temperature, the product is absorbed onto a highly acidic ion exchange resin. Elution with 2% aqueous pyridine solution and freeze drying gave the crude product which was further purified on a Sephadex L H-20 chromatographic column using methanol. The data from and hP NMR spectra and from the IR / infrared / spectrum are consistent with the structure. A satisfactory fuel analysis was obtained for the monohydrate. After silylation, the mass spectrum showed peaks at 572 (M ⁺ ), 557 (M ⁺ -15) and at 455 (M + - CO2) Me ₃ (m / e).

Example 143. N-p-Carboxy-3- (4-imidazolyl) propyl / L-alanyl-L-propion.

N- (1-carboxy-3- (4- (L) -benzylimidazolyl) propyl) -L-alanyl-L-proline, obtained by the method described in Example 142, is hydrogenated at 40 gauge pressure (pounds).

- 99 2 inch water equivalent weight 5H-en paladin / charcoal mixture of acetic acid / water at a ratio of 3 to 1 psi-pat-with ^G ' ^{/ 3} C' 2 pg. . Cf ?,. · Defending Potslips with 'Iltrips of .Iltrato' soup product

Strongly acidic

a resin, followed by an elution with a 2-aqueous aqueous solution of pyridine and freeze-drying. ! o-insist: but purification is carried out on a Secodex L H-20 column using methanol. The proton GF spectrum and the thin-layer chromatography, but the toduct showed that the bezil group was removed to obtain the title compound, had a peak peak of / ♦ - 10 / at m / e 320.

EXAMPLE 144. N- (1-Carboxyl-3-yl) -benzylimidazolyl (in poopyl) -L-lysyl-L-proline and N- (1- (4-yl) -azolyl) -

PROPYL / -I-lysyl-L-PROLIN.

The two title compounds were prepared in the same manner as N- (1-narboxy-3- (4- (1-benzylimidazolyl)) poopyl-N-allonyl-L-pooline (Example 142) and N- (L-carboxy) -A-iadazolyl (poopyl) -N-alanyl-L-proline by substitution with L-alan / l-L-apiene with Et- (R) -L-lisyl-L- (R) -H-H and removal of tert. 3C tri trifluoroacetic acid at tapeosturs 0 ^and C for 3 h after condensation.

Example 145. N- / I. -etonsikabonyl-5- (4- (1-sensylimidazolyl) propyl) -L-alanyl-L-proline and N- (L-ethoxynorbonyl) -? - (4-yl): idazolyl / propyl-L-alonyl-L - Proline.

The two compounds mentioned in the title were prepared in the same way as N- (1-coboxo-5- (4-) l -benzylimidazolyl // poopyl) -L-eleonyl-L-proline (primer 142).

BAD ORIGINAL

100 N- (1-carboxy-3 (4-imidazolyl) propyl) -L · -alanil-L-proline (Example 143) using 4- (4- (1-benzylimidazolyl) ethyl ester // - 2- ketobutyric acid and condensation in ethanol (in the presence of molecular sieves 4A) instead of in water.

REFERENCES

1. US 4129571

2. US 4154960

3. US 4052511

4. Cushman D.W. et al., Biochemistry 16, 5484, 1977

5. PlQUILLOUD Y., A. REINHARZ AND M. ROTH, BlOCHEM BlOPHYS.

Acta 206, 136, 1970

6. Weeks J.R. and J.A. Jones, Proc. Soc. Exp. Biol. Med., 104, 646, 1960

7. Koletsky S. et al., Proc. Soc. Exp. Biol. Med., 125, 96, 1967

Claims (82)

Compound of Formula II p NH ci II in which they are the same or different and denote hydroxy, lower alkoxy,, dinisalkylamino lower alkoxy, acylamino lower alkoxy, acyloxy lower alkoxy aryloxy, arylalkylalkyloxy, substituted aryloxy or substituted arylloxyalkyl, methylamino, methylsilyl, methylsilyl, methylsilyl, methylsilyl, methylsulfonyl, , arylalkylamino, R-] represents hydrogen, alkyl containing 1 to 20 or hydroxyamino; carbon atoms, including branched, cyclic and unsaturated alkyl groups; substituted lower alkyl, with the substituent being halo, hydroxy, lower alkoxy, aryloxy, amino, lower alkylamino, dinisalkylamino, acylΜΤϊ idino, imidazolyl, indolyl, mercapto, lower alkoxycarboxyl, aridoxyl, carboxyl, aridoxylcarboxyl substituted phenyl wherein the substituent is lower alkyl, lower alkoxy or halo; arilnisshalkil or heteroarilnisshalkil, arilnisshalkenil or heteroarilnissholkenil substituted arilnisshalkil substituted heteroarilnisshalkil substituted arilnisshalkenil or substituted heteroarilnissholkenil, the substituent is halo or dihalo, loweralkyl, hydroxy, loweralkoxy, amino, aminomethyl, acylamino, dinisshalkilamino, loweralkylamino, carboxyl, haloloweralkyl, cyano or sulfonamido; arylalkyl or heteroarylalkyl substituted on the alkyl moiety by amino or acylamino groups; RUB2 and Ry denote hydrogen or alkyl; means hydrogen, lower alkyl, phenyl lower alkyl, amino phenyl lower alkyl, hydroxy phenyl lower alkyl, hydroxy lower alkyl, acetyl lower alkyl, lower alkyno , indolyl lower alkyl, mernaptly lower alkyl and lower alkylthio lower alkyl; R 4 represents hydrogen or lower alkyl; R1 represents hydrogen, lower alkyl, phenyl, phenyl lower alkyl, hydroxy phenyl lower alkyl, hydroxy lower alkyl, amino lower alkyl, guanidino lower alkyl, imidosolyl lower alkyl, indolyl lower alkyl, mercenyl alkyl, mercapto alkyl, mercapto alkyl; and R5 may be linked together to form an alkylene fat of 2 to 4 carbon atoms, an alkylene bridge of 2 or 3 carbon atoms and one sulfur atom, an alkylene bridge of 3 or 4 carbon atoms containing a double bond, or an alkylene bridge of the type of the foregoing substituted by hydroxy lower alkoxy, lower alkyl or dinisyl alkyl and their pharmaceutically acceptable salts, wherein said aryl is a member of the group consisting of phenyl or naphthyl, and said heteroaryl is a member of the group consisting of pyridyl, thienyl, furyl, indolyl, benzthienyl, imidosis Lille or tiazrlil. 2. A compound according to claim 1, wherein the carbon atom bearing the substituent is a B5 configuration and the other absolute configurations belong to the L-amino acids. 3. A compound of formula 0 I / , ^R 1 * c ( ^J * 5 0 N And? - with -  C -NH I -CH -c II -N - - C - 1 C - Rg ^R 2 0 ^? 7 wherein R and Rg may independently represent hydroxy, lower alkoxy lower alkenyloxy, aryl lower alkyloxy, amino, dinisalkylomino lower alkoxy, acylamino lower alkoxy or acyloxy lower alkoxy; R < 1 &gt; denotes hydrogen, alkyl containing I to 20 carbon atoms, including branching, cyclic and unsaturated alkyl groups; substituted lower alkyl, the substituent being halo, hydroxy, lower alkoxy, aryloxy, amino, lower alkylamino, dinisalkylamino, acylamino, arylamino, guanidino, imidazoyl, indolyl, mercapto, loweralkylthio, carboxycarboxycarboxylcarboxylcarboxyl wherein the substituent is lower alkyl, lower alkoxy or halo; arilnisshalkil or heteroarilnissh alkyl arilnisshalkenil or heteroarilnissh alkenyl, substituted aryl, lower alkyl, substituted heteroarilnisshalnil substituted arilnisshalkenil or substituted heteroarilnisshalkenil in retard the substituent is halo, dihalo, loweralkyl, hydroxy, loweralkoxy, amino, aminomethyl, acylamino, dinisshalkilamino, loweralkylamino, carboxyl, haloalkyl cyano or sulfonamido, arylalkyl, or heteroarylalkyl substituted on the alkyl moiety by amino or acylamino groups; R? and R ₇ is hydrogen; R ₃ represents lower alkyl amino lower alkyl imidazolyl lower alkyl, halo lower alkyl; 5 are linked together to carbon R ₄ and R containing from 2 to 4 the formation of an alkylene atom or an alkylene bridge, BRIDGE containing 2 or 3 carbon atoms, and one sulfur or alkylene atom containing from 3 to 4 carbon atoms containing a double bond _/ or alkylene bridge of the above type substituted with hydroxy, lower alkoxy or lower alkyl, or pharmaceutically acceptable salts, wherein said aryl is selected from the group consisting of phenyl or naphthyl, and said hetero aryl is selected from the group consisting of pyridyl, thienyl, furyl, indolyl, benzthienyl, imidazoyl, or thiazolyl. A compound according to claim 3 wherein the carbon - 4 atom but bearing the substituent R- ^ e in 6 configuration and the other absolute configurations belong to the L-amino acids. The compound of claim 3, wherein R 1 and R 8 are linked together by both the carbon and nitrogen atoms. to which they are attached form a ring of formula ΟΰΰΖζ wherein Y represents CH ^, a sulfur atom or CH-OCH ₃ and Rp Rg and R? have the meanings given in claim 3. Compounds according to claim 5, wherein the carbon bearing the denominator has the S configuration and the other absolute configurations belong to the L-amino acids. A compound of formula 0 % ^R 4 is ₅ o 1 ' I 1 1 II R - C -C -NH-CH - - With -N II -C-C -Rg 0 ^R 7 wherein R and Rg may independently represent hydroxy, lower alkoxy, lower alenyloxy, aryl lower alkyloxy, amino, dinisalkylamino lower alkoxy, acylamino lower alkoxy or acyloxy lower alkoxy; 1? 1 means alkyl containing 1 to 8 carbon atoms, substituted lower alnyl, the olyl group having 1 to 5 carbon atoms, and the substituent being amino, orythio, oryloxy, arylamino, arylalkyl or heteroarylalkyl, with the alkyl moiety having 1 to 3 carbon atoms , substituted arylalkyl or heteroarylalkyl, the alkyl moiety having 1 to 3 carbon atoms and the substituent (s) being halo, respiratory, amino, aminoalkyl, hydroxy, lower alkoxy or lower alkyl; ^and ^ 7 represent hydrogen; R ₃ represents lower alkyl or amino lower alkyl; R 2 and R 8 are bonded together with the carbon and nitrogen atom to which they are attached to form a ring of formula wherein Y represents CH ₂ , sulfur atom or CH-OCH ₃ or its pharmaceutically acceptable salts, wherein said aryl is selected from the group consisting of phenyl or naphthyl, and said heteroaryl is selected from the group consisting of pyridyl, thienyl, furyl, indolyl, benzalkenyl, IMIDO30IL or TIO30LIL. Compounds according to claim 7, wherein the carbon atom bearing the substituent R1 is in the S configuration and the other absolute configurations belong to the L-amino acids. 9. Compound of Formula ^{0 R} 1 ^R 3 ^R 4 ^R 5 i · I II * R-CC-NH-CH-CNCCR ₆ 'II R ₂ 0R in which R and Rg independently represent hydroxy, lower alkoxy, arylalkyloxy; R? and Ry denotes hydrogen; R ₃ denotes methyl, aminonylalkyl; R ^ and Rg are bonded together with the carbon and nitrogen atom to which they are attached to form proline, 4-thiaprolin or 4-methoxyproline, and R 4 represents alkyl containing from 1 to 8 carbon atoms, substituted lower alkyl, the alkyl group having 1 to 5 carbon atoms and the substituent being amino, arylthio, aryloxy, arylalkyl or heteroarylalkyl, the alkyl moiety having 1 to 3 carbon atoms, substituted arylalkyl or heteroarylalkyl, wherein the alkyl group has 1 to 3 carbon atoms and the substituent (s) are halo, respiratory, amino, aminoalkyl, hydroxy, lower alkoxy or lower alkyl and its pharmaceutically acceptable salts, said aryl being selected from the group consisting of from phenyl or naphthyl, and said heteroaryl is selected from the group consisting of pyridyl, thienyl, furyl, indolyl, benzthienyl, imidazoyl or thiazolyl. Compounds according to claim 9, wherein the carbon bearing the substituent is in the 5 configuration and the other absolute configurations belong to the L-amino acids. A compound according to claim 9, which is N- (1-carboxy-3-phenylpropyl) -1-3-alanyl-L-proline. A compound according to claim 9, which is N- (N-ethoxycarbonyl-3-phenylpropyl) -L-alanyl-L-proline. The compound of claim 9, which is N- (1-ethoxycarbonyl-4-methylpentyl) -L-alanyl-L-proline. A compound according to claim 9, which is N-1-carboxy-5-aminopentyl-L-alanyl-L-proline. The compound of claim 9, which is N-N- (1-carboxy-3-phenylpropyl) -L-lysyl-L-proline. A compound according to claim 9 which is N-N- (1-ethoxycarbonyl-3-phenylpropyl) -L-lysyl-L-proline. A compound according to claim 9, which is N-N- [1-carboxy-3- (3-indolyl) -propyl] -L-lysyl-L-proline. A compound according to claim 9, which is L1- [1- (1-carbony-3/4-chlorophenyl) propyl] - L-lysyl-L-proline. A compound according to claim 9, which is N- (1- (1-carboxy-2-phenylthioethyl) - L-lysyl-L-proline. A compound according to claim 9, which is N- (N-carboxy-3- (4-chlorophenyl) propyl) N-lysyl-trans-4-methoxy-L-proline. A compound according to claim 9, which is N - [1- (1-carboxy-5-min) propenyl] -1'-lysyl-L-proline. A compound according to claim 9, which is N- [1- (1-carboxy-3-phenylpropyl) -L-ornyl-1 H -proline. A compound according to claim 9, which is ethyl N- (1-ethoxycarbonyl-3-phenylpropyl) -L-alanyl-L-proline hydrochloride. A compound according to claim 9 which is N- (1- (ethoxycarbonyl) -3- (4-imidazolyl) propyl) -L-alanyl-L proline. A compound according to claim 9 which is b1- (1- (1-carbonyl-3- (4-imidazolyl) propyl) -L-lysyl-1 '-proline. 26. The compound N- (R) -carboxy-3-phenylpropyl) -L alanyl-L-proline. 27. The compound N- (N) -5-ethoxycarbonyl-3-phenyl-propyl / L-alanyl-L-proline. 28. The compound N- (1/5) -ethoxycarbonyl-3-phenyl-propyl / L-alanyl-L-proline maleate. 29. The compound NV- [I (S) -carboxy-3-phenyl-propyl] -L lysyl-L-proline. 30. The compound ethyl N- (1- (5) -ethoxycarbonyl-3-phenylpropyl) -L-alanyl-L-proline hydrochloride. 31. The compound N-2- (N) -5-ethoxycarbonyl-3-phenylpropyl / L-lysyl-L-proline. 32. A pharmaceutical composition for the treatment of hypertension, which includes a pharmaceutically acceptable carrier and a formulation, a pharmaceutically acceptable carrier, and a pharmaceutically effective amount of a compound of formula | RCC-NH-CH-CNCC-Rr> II I ⁶ ξ ₂ 0 r ₇ in which R and Rg may be the same or different and denote hydroxy lower alkoxy, lower alkenyloxy, dinisalkylamino lower alkoxy, acylamino lower alkoxy, acyloncy lower alkoxy, arylons, aryl lower alkyloxy, substituted aryloxy or substituted arylxy, substituted or methyloxy, substituted methyloxy, , lower alkyldinylalkylamino. amino, arylalkylamino, hydroxyamino; R. J represents hydrogen, alkyl containing I to 20 carbon atoms, including branched, cyclic or unsaturated alkyl groups; substituted lower alkyl, such as. the substituent is halo, hydroxy, lower alkoxy, aryloxy, amino, lower alkylamino, dinisalkylamino, acylamino, arylamino, guanidino, imidazolyl, indolyl, mercapto, lower alkylthio, arylthio, carboxy, carboxamido, carboxyamido, carboxyamido, carboxyamido, carboxyamido, carboxyamido, carboxyamido, carboxyamido, carboxy , lower alkoxy or halo, arylalkyl or heteroarylalkyl, arylalkenyl or heteroarylalkenyl, substituted arylalkyl, substituted heteroarylalkyl, substituted arylalkenyl, or substituted heteroarylalkenylalkenyl, or a respirator, lower alkyl, hydroxy, lower alkoxy, amino, aminomethyl, acylamino, dinisalkylamino, lower alkylamino, carboxyl, halonylalkyl, cyano or sulfonamido; arylalkyl or heteroarylalkyl substituted on the alkyl moiety with amino or acylamino; and R ₇ is hydrogen or lower alkyl; R ₃ represents hydrogen, lower alkyl, fenilnisshalkil, ominometil phenyl nisshalnil, hydroxyphenyl lower alkyl, hidroksinisshalkil, atsetilaminonissh alkyl, acylamino lower alkyl, amino lower alkyl, dimetilamino'nissh (alkyl, guanidino lower alkyl, halo | lower alkyl, imidazalil lower alkyl, indolyl 'lower' alkyl, mercanto) lower [alkyl, lower alkylthio lower alkyl; means hydrogen or lower alkyl; 1-5 denotes hydrogen, lower alkyl, phenyl, phenyl lower alkyl, hydroxyphenyl lower alkyl, hydroxy lower alkyl, amino lower alkyl, guanidino lower alkyl, imidazolyl lower alkyl, indolyl lower alkyanyl mercylalkyl or alkali alkylene alkyl mercallyalkyl; and bonded together with the carbon and nitrogen atoms to which they are attached form an alkylene bridge containing 2 to 4 carbon atoms, an alkylene bridge containing 2 or 3 carbon atoms and one sulfur atom, an alkylene bridge containing 3 to 4 carbon atoms containing a double bond or alkylene bridge of the above type, substituted by hydroxy, lower alkoxy or lower allyl and its pharmaceutically acceptable salts, but said aryl is selected from the group consisting of phenyl or naphthyl and said heteroaryl is selected from the group consisting of feasts yl, thienyl, furyl, indolyl, benzthienyl, imidazoyl or thiazolyl. 33. A pharmaceutical composition for the treatment of high blood pressure comprising a pharmaceutically acceptable carrier and a pharmaceutically effective amount of a compound of formula 0 * 1 R ₃ 1 ^ 5 II I I 11 RCC-NH-CH-C'NCC-Rzi li I and ^b 1 ^ 2 θ in which P and Rg are, independently of one another, hydroxy, lower alkoxy, lower alkynyloxy, aryl lower alkoxy, amino, dinisalkyl amino lower alkoxy, acylamino lower alkoxy or acylonyl lower alkoxy; means alkyl containing 1 to 8 carbon atoms, substituted lower alkyl, the alkyl group having 1 to five carbon atoms and the substituent being amino, arylthio, aryloxy, or arylamino, arylalkyl or heteroarylalkyl, with the alkyl moiety having 1 to 3 carbon atoms, arylalkyl or heteroarylallyl, the alkyl moiety having 1 to 3 carbon atoms and the substituent (s) are halo, respiratory, amino, aminoalkyl, hydroxy, lower alkoxy or lower alkyl; R ₂ and Ry denote hydrogen; 1? ₃ means lower alkyl or amino lower alkyl; and are attached together with the carbon and nitrogen atom to which they are attached and form a ring of formula wherein Y represents CH ₂ , sulfur atom or CH-OCH ₃ or its pharmaceutically acceptable salts, wherein said aryl is selected from the group consisting of of phenyl or naphthyl and said heteroaryl is selected from the group consisting of pyridyl, thienyl, vindolyl benzthienyl, imidazoyl or thiazolyl. 34. A pharmaceutical composition for the treatment of hypertension but comprising a pharmaceutically acceptable carrier and a pharmaceutically effective amount of a compound of the general formula 0 R-> R ~ Rd Rc to ι ι ³ ) ⁴ P RCC-NH-CH-CNCCR, ¹ II I I. ^{6 R} 2 0 P _? O in which R 6 and R 8 independently represent hydroxy, lower alkoxy, arylalkyloxy; p _and denote hydrogen; R ₃ denotes methyl, aminonylalkyl; R1 and R8 are attached together with the carbon and nitrogen atom to which they are attached and form proline, 4-thiaprolin, 4-methoxyproline and R-Ι means alkyl containing 1 to 8 carbon atoms, substituted lower alkyl, but the alkyl group has one to five carbon atoms and the substituent is amino, arylthio, aryloxy; arylalkyl or heteroarylalkyl, wherein the alkyl moiety contains 1 to 3 carbon atoms, substituted arylalkyl or heteroarylalkyl, the alkyl moiety containing 1 to 3 carbon atoms, and the substituent (s) is halo, breathing, amino, aminoalkyl, hydroxy, lower or lower alkyl pharmaceutically acceptable salts, wherein said aryl is selected from the group consisting of phenyl or naphthyl, and said heteroaryl is selected from the group consisting of pyridyl, thienyl, furyl, indolyl, benzthienyl, imidazoyl or thiazolyl. The composition of claim 34, wherein said compound is N- [1- (1-carboxy-3-phenylpropyl) -L-alanyl-L-proline. The composition of claim 34, wherein said compound is N- (1-ethoxycarbonyl-3-phenylpropyl) -N-alanyl-L · proline. The composition of claim 34, wherein said compound is N- [- (1-ethoxycarbonyl) -4-methylpentyl-L-alanyl-L proline. The composition of claim 34, wherein said compound is N- (1-carboxy-5-aminopentyl) -L-alanyl-L-proline. 39. The composition of claim 34, wherein said compound is N ₀ / - / 1-carboxy-3-phenylpropyl / - L-lysyl-L · proline. The composition of claim 34, wherein said compound is N - N - (1-ethoxycarbonyl-3-phenylpropyl) -L-lysylL-proline. 41. The composition of claim 34, wherein said compound is 1M- _with / - / 1-carboxy-3- / 3-indolyl / propyl / -L lysyl L -proline. The composition of claim 34, wherein said compound is N- _O / - / N-καρ6οκοπ-3-4-chlorophenyl / propyl-L-lysyl-L proline. The composition of claim 34, wherein said compound is b1 - [- (1-carboxy-2-phenylthioethyl) - L-lysyl-L · -proline. The composition of claim 34, wherein said compound is N-N- [1-carboxy-3- (4-chlorophenyl) propyl] -L-lysyltrans-4-methoxy-L-proline. 45. The composition of claim 34, wherein said compound is N- _is / - / 1-carboxy-5-ominopentil / -L-lysyl-L-proline. 46. The composition of claim 34, wherein said compound is N- _er / - / 1-narbonsi-3-phenylpropyl / -A -ornitil-L-proline. The composition of claim 34, wherein said compound is ethyl N- (1-ethoxynorbonyl) -3-phenylpropyl] -L-alonylL-proline hydrochloride. 48. The composition of claim 34, wherein said compound is N- _c / - / 1-carboxy-3-phenylpropyl / -L -ornitil-L · proline. The composition of claim 34, wherein said compound is ethyl b1- (1-ethoxycarbonyl-3-phenylpropyl) -L-alanylb-proline hydrochloride. The composition of claim 34, wherein said compound is N- (1-ethoxycarbonyl-3- (4-imidazolyl) propyl) -L-alanyl-L-proline. The composition of claim 34, wherein said compound is N- (1-carboxy-3) -4-imidazolyl (propyl) -L-lysyl- L · -proline. The composition of claim 34, wherein said compound is N- (N) -carboxy-3-phenylpropyl] -L-alonyl-L proline. The composition of claim 34, wherein said compound is N- (N) -5-ethoxycarbonyl-3-phenylpropyl] -L-alanyl- L · -proline. The composition of claim 34, wherein said compound is N- (1/5) -ethoxycaronyl-3-phenylpropyl] -L-alanyl- L · -proline maleate. The composition of claim 34, wherein said compound is N - [(N) -5-carboxy-3-phenylpropyl] -L-lysyl-L-proline. The composition of claim 34, wherein said compound is ethyl N- (N) -ethoxycarbonyl-3-phenylropyl) -L alanyl-L-proline hydrochloride. The composition of claim 34, wherein said compound is N - [- (1/5) -ethoxycarbonyl-3-phenylropyl] -L-lysylL-proline. 58. A method of treating hypertension, which comprises administering to a patient a body a pharmaceutically effective amount of a compound of formula 0 and * 5 0 II R-C- -C-NH 1 -from - with - II - N · - C - 1 C -R ₆ * 2 0 ^R 7 in ostrich R and Rg are the same or different and denote hydroxy, lower alkoxy, lower alkenyloxy, dinisalkylamino lower alkoxy, acylamino lower alkoxy, acyloxy lower alkoxy, aryloxy, aryl lower alkyloxy, substituted aryloxy or substituted aryl methyloxyalkyl; amino, lower alkylamino, dinisalkylamino, aryl lower alkylamino or hydroxyamino; means hydrogen, alkyl containing 1 to 20 carbon atoms, including branched, cyclic or unsaturated alkyl; substituted lower alkyl, wherein the substituent is halo, hydroxy, lower alkoxy aryloxy, amino, lower alkylamino, dinisalkylamino, acylamino, arylamino, guanidino, imidazolyl, indolyl, mercapto, lowershalkynoxycarboxyl, carboxyl, carboxyl, carboxyl, carboxyl wherein the substituent is lower alkyl, lower alkoxy or halo; arilnisshalkil or heteroarilnisshalkil, arilnisshalkenil or heteroarilnisshalkil substituted arilnisshalkil substituted heteroarilnisshalkil substituted arilnisshalkenil or substituted heteroarilnisshalkenil, wherein the substituent is halo or dihalo, loweralkyl, hydroxy, loweralkoxy, amino, aminomethyl, acylamino, di-lower-amino, nisshalkilomino, carboxyl , halo lower alkyl, cyano or sulfonamido; arylalkyl or heteroarylalkyl substituted on the alkyl moiety by amino or acylamino; R2 and Ry denote hydrogen or lower alkyl; R ₃ represents hydrogen, lower alkyl, phenyl lower alkyl, aminomethyl phenyl lower alkyl, hydroxyphenyl nisshalnil, hidroksinisshalkil, acetylamino loweralkyl, atsilaminonisshalkil, amino loweralkyl, dimethylamino lower alkyl, guanidino lower alkyl, halo lower alkyl, imidazolyl lower alkyl, indolyl lower alkyl, mercapto lower alkyl and lower alkylthio lower alkyl; R 4 represents hydrogen or lower alkyl; ^ 5 means hydrogen, lower alkyl, phenyl, phenyl lower alkyl, hydroxyphenyl lower alkyl, hydroxy lower alkyl, amino lower alkyl, guano dinino lower alkyl, imidazolyl lower alkyl, indolyl lower alkyl, mercapto lower alkyl, lower alkylthio lower alnyl; ^R 4 and R ₅ can be linked together to form alkylene bridge containing 2 to 4 carbon atoms, alkylene bridge containing 2 to 3 carbon atoms and one sulfur atom, alkylene bridge containing 3 to 4 carbon atoms and a double bond or alkylene bridge of the above type substituted by hydroxy , lower alkoxy or lower alnyl, and pharmaceutically acceptable salts thereof, wherein said aryl is selected from the group consisting of phenyl or naphthyl, and said heteroaryl is selected from the group consisting of pyridyl, thienyl, furyl, indolyl, benzylthienyl , imidazolyl or thiazolyl. 59. A method of treating hypertension, which involves administering to a patient a pharmaceutically effective amount of a compound of formula 0 tl in s R - C - - with - 1 NHCH - with - 1 » - N - - C - 1 C - R <11 ⁶ In 0 ^R 7 0 wherein R and Rg independently represent hydroxy, lower alkoxy, arylalkyloxy, R.] means alkyl having 1 to 8 carbon atoms, substituted lower alkyl, wherein the alkyl group has from one to five carbon atoms and the substituent is amino, arylthio or aryloxy, arylalkyl or heteroarylalkyl, the alkyl moiety containing from The 1 to 3 carbon atoms, substituted arylalkyl or heteroarylalkyl alkyl moiety contains 1 to 3 carbon atoms and the substituents (s) are halo, respiratory, amino, aminoalkyl, hydroxy, lower alkoxy, lower alkyl; 1? 2 and Ry denote hydrogen; R ₃ denotes methyl, aminonylalkyl; and may be coupled together with the carbon and nitrogen atom to which they are attached to form proline, 4-thialrolin, or 4-methoxyproline, or pharmaceutically acceptable salts thereof, wherein said aryl is selected from the group consisting of phenyl or naphthyl, and said heteroaryl is a member selected from the group consisting of pyridyl, thienyl, furyl, indolyl, benzthienyl, imidazoyl or thiazolyl. The method of claim 59, wherein said compound is N- (1-carboxy-3-phenylpropyl) -L-alanyl-L proline. The method of claim 59, wherein said compound is b1- (1-ethoxycarbonyl-3-phenylpropyl) -11-alanylL-proline. The method of claim 59, wherein said compound is N- (1-ethoxycarbonyl-4-methylpentyl) -N-alanyl-L-proline. 63. The method of claim 59, wherein said compound is N- (1-carboxy-5-aminopentyl) -1 and -alanyl-b proline. 64. The method of claim 59, wherein said compound is N - [- (1-carboxy-3-phenylpropyl) -L-lysyl-L · - proline. 65. The method of claim 59, wherein said compound is N-N-N-ethoxycarbonyl-3-phenylpropyl] -L-lysyl] N -proline. 66. The method of claim 59, wherein said compound is N- _c / - / 1-carboxy-3- / 3-indolyl / propyl / -L -lizilL proline. 67. The method of claim 59, wherein said compound is N- _c / - / 1-carboxy-3- / 4-chlorophenyl / propyl / -L lysyl B -proline. 68. The method of claim 59, wherein said compound is N- _O- (1-carboxy-2-phenylthioethyl) -L-lysyl- L -proline. The method of claim 59, wherein said compound is 1 N - [- (1-carboxy-3- (4-chlorophenyl) propyl) -L-lysyltrans-4-methoxy-L-proline. The method of claim 59, wherein said compound is N- [1- (1-carboxy-5-aminopentyl) -1-lysyl-L proline. The method of claim 59, wherein said compound is b1- S - (N-carboxy-3-phenyl propyl) -N-ornyl-L proline. The method of claim 59, wherein said compound is ethyl N71-ethoxycarbonyl-3-phenylpropyl] -L-alanyl L-proline hydrochloride. The method of claim 59, wherein said compound is N- [1- (1-carboxy-3-phenylpropyl) -L-ornyl-L proline. 74. The method of claim 59, wherein said compound is ethyl N- (1-ethoxycarbonyl-3-phenylpropyl) -L-alanylL-proline hydrochloride. The method of claim 59, wherein said compound is N- (1- (ethoxycarbonyl) -3- (4-imidazolyl) propyl) -L-alanyl-L-proline. 76. The method of claim 59, wherein said compound is b1- (1-carboxy-3- (4-imidazolyl) propyl) -L-lysylL-proline. The method of claim 59, wherein said compound is N- (N) -5-carboxy-3-phenylpropyl) -L-alanyl-L proline. The method of claim 59, wherein said compound is N- (R) -ethoxycarbonyl-3-phenylpropyl] -L-alanyl-1 '-proline. 79. The method of claim 59, wherein said compound is N- (1 / S) -oxycarbonyl-3-phenylpropyl] -alanil-L · -proline maleate. The method of claim 59, wherein said compound is N- (N) -carboxy-3-phenylpropyl] -L-lysyl-L · proline. 81. The method of claim 59, wherein said compound is ethyl N- (N) -ethoxycarbonyl-3-phenyl-propyl) -L alanyl-L · -rolinate hydrochloride. 82. The method of claim 59, wherein said compound is N - [(N) -5-ethoxycarbonyl-3-phenylpropyl] -L lysyl-L · -proline.