BG60368B2 - Derivatives of aminoacids of antihypertension activity - Google Patents

Abstract

The invention includes methods for making carboxyalkyl dipeptide derivatives and related compounds that are used as angiotensin converting enzyme inhibitors and as antihypertensive and other pharmaceutical preparations containing these carboxyalkyl dipeptide compounds in combination with another compound with antihypertensive and / or diuretic action.

Description

U.S. Patent Nos. 4,129, 571, and 4,154,960 disclose substituted acyl amino acid derivatives for use as inhibitors of the angiotinin converting enzyme. In particular, these compounds are mercaptosubstituted acylamino acids and their derivatives, incl. the clinically effective antihypertensive compound captopril O-3-mercapto-2methylpropanoyl-1-proline.

Said compounds of the prior art are not dipeptide derivatives, such as the compounds of the invention. In addition, the prior art compounds contain a sulfhydryl substituent or derivative thereof, and the compounds of the invention do not contain such a substituent. The dipeptide compounds of the invention are unusual dipeptides whose terminal N is attached to a carboxymethylene group, which is preferably further substituted by a methyl group. In addition, the carboxyl group (s) may also be converted to derivatives such as esters, amides, salts. In fact, the compounds of the invention are hybrids formed by binding α-amino acids to dipeptides via a nitrogen atom that is common to these two-part structures. Such a structural arrangement is unusual in the field of synthetic and natural peptides and is characteristic of the mercaptoacyl type of groups in the prior art described above.

U.S. Patent No. 4,052,511 lists Ncarboxylalkanoylamino acids, which are used as inhibitors of the angiotensin converting enzyme. Because the compounds of the invention are dipeptide derivatives, in the formal sense they can be regarded as being related to any of the compounds described in US Patent No. 4,052,511. When one of the methylene groups is replaced by an amino group of the invention, compounds of very high efficiency are obtained. For example, preferred compounds of the invention may be administered in low doses, e.g., 2.5 mg per patient per day compared to the lowest dose of 1 mg / kg per day for preferred compounds of US Patent No. 4,052,511, corresponding to 60 mg / day of percent, with an average patient weight of about 68 kg.

Description of the invention

The invention relates to carboxyalkyl dipeptides and derivatives thereof, which are used as inhibitors of the conversion enzyme and as antihypertensive agents. The compounds of the invention can be represented by the following formula

About r1 r3 r4 r5 ο

II I I I I I II

R-C-C-NH-CH-C-N-C-C-R6

L · III,

R 2 OR ⁷ j wherein R and R ⁶ are the same or different and are hydroxy, lower alkoxy, lower alkoxy, lower dialkylamino lower alkoxy (dimethylaminoethoxy), acylamino lower alkoxy (acetylaminoethoxy), acyloxy lower alkoxy (pivaloyloxyloxymethyl) , aryl lower alkoxy such as benzyloxy, substituted aryloxy or substituted aryl alkoxy, the substituent being methyl, halogen or methoxy, amino, lower alkylamino, lower dialkylamino, hydroxyamino, aryl lower alkylamino such as benzylamino;

R * is hydrogen, alkyl of from 1 to 20 carbon atoms, including side and cyclic and unsaturated (such as allyl) alkyl groups, substituted lower alkyl, the substituent being halogen, hydroxy, lower alkoxy, aryloxy such as phenoxy, amino, dinisulf alkylamino, acylamino, such as acetamino and benzamido, arylamino, guanidino, imidazolyl, indolyl, mercapto, lower alkylthio, arylthio, such as phenylthio, carboxy or carboxamido, carbonisalkoxy, aryl, such as phenyl or naphthyl, phenyl or naphthyl alkyl, lower alkoxy or halogen, aryl lower al carbyl, aryl lower alkenyl, heteroaryl lower alkyl or heteroaryl lower alkenyl, such as benzyl, styryl or indolyl ether, substituted aryl lower alkyl, substituted aryl lower alkylene, substituted heteroaryl lower alkyl, or substituted heterostenyl, hetero-esteryl esterallyl esterallyl esteryl ester; lower alkyl, hydroxy, lower alkoxy, amino, aminomethyl, acylamino (acetylamino or benzoylamino) dinisyl alkylamino, lower alkylamino, carboxyl, halogenated lower alkyl, cyano or sulfonamido, aryl lower alkylaryl alkylene alkyl by amino or acylamino (acetylamino or benzoylamino);

R ² and R ⁷ are the same or different and are hydrogen or lower alkyl;

R ³ is hydrogen, lower alkyl phenyl lower alkyl, aminofenilmetil lower alkyl, hydroxyphenyl lower alkyl, acylamino lower alkyl (such as benzoylamino lower alkyl, acetylamino lower alkyl), amino lower alkyl, dimethylamino lower alkyl, halogenated lower alkyl, indolyl lower alkyl, mercapto lower alkyl, lower alkylthio lower alkyl;

R ⁴ is hydrogen or lower alkyl;

R ⁵ is hydrogen or lower alkyl, phenyl, phenyl lower alkyl, hydroxyphenyl lower alkyl, hydroxy lower alkyl, amino lower alkyl, guanidino lower alkyl, imidazolyl lower alkyl, indolyl lower alkylsilylalkyl alkyl mercitalkyl, mercapto alkyl

R ⁴ and R ³ may be linked together to form an alkylene bridge of 2 to 4 carbon atoms, an alkylene bridge of 2 to 3 carbon atoms and one sulfur atom, an alkylene bridge of 3 to 4 carbon atoms containing one double bond or an alkylene bridge, as above substituted by hydroxy, lower alkoxy, lower alkyl or lower dialkyl; and their pharmaceutically acceptable salts.

The lower alkyl or lower alkenyl groups, unless otherwise noted, described in any of the embodiments of the invention contain hydrocarbon radicals having one to six carbon atoms, for example methyl, ethyl, propyl, isopropyl, butyl, isobutyl, n- butyl, pentyl, isopentyl, hexyl or vinyl, allyl, butenyl and the like. The arylalkyl groups described in each of the above variants have from one to four carbon atoms in their alkyl moiety and include, for example, benzyl, p-methoxybenzyl and the like. Halogen means chlorine, bromine, iodine or fluorine. Aryl in each of the embodiments, unless otherwise indicated, is phenyl or naphthyl. Heteroaryl groups when they occur include, for example, pyridyl, thienyl, furyl, indolyl, benzthienyl, imidazolyl and thiazolyl.

Substituted lower alkyl moieties R, R and R are illustrated by groups such as

'I

BUT-CH _? -, HS-CH ₂ -, H ₂ N- (CH 2) ₄ -, CH ₃ -S- (CH 2) ₂ -, H ₂ N- (CH 2) ₃ and others.

When R ⁴ and R ⁵ are bonded via the carbon and nitrogen atom to which they are attached, they form a 4- to 6-membered ring which may contain one sulfur atom or one double bond. Preferred rings have the following formula

wherein Y is CH ₂ , S or CHO ₃ .

Preferred are those compounds of formula I in which

R and R ⁶ can be independently and independently hydroxy, lower alkoxy, lower alkoxy, aryl lower alkoxy, amino, lower dialkylamino lower alkoxy, acylamino lower alkoxy, acyloxy lower alkoxy, the substituent being methyl, halogen or methoxy;

R ² and R ⁷ are hydrogen;

R ³ is lower alkyl, amino lower alkyl, imidazoyl lower alkyl, halogen lower alkyl;

R ⁴ and R ⁵ are bonded to form the preferred said rings in which Y is CH ₂ , S or CH-OCH ₃ ;

R ¹ is as defined above.

Even more preferred are those compounds of formula I, wherein R ¹ is also alkyl having from 1 to 8 carbon atoms; substituted lower alkyl wherein the alkyl group has 1 to 5 carbon atoms and the substituent is amino, arylthio, aryloxy or arylamino, arylalkyl or heteroarylalkyl, wherein the alkyl moiety has 1 to 3 carbon atoms such as phenethyl or indolylethyl or substituted arylalkyl or substituted naphthyl (lower alkyl) and substituted heteroarylalkyl, wherein the alkyl groups have 1 to 3 carbons and wherein the substituent (s) is / are halogen, dihalogen, amino, aminoalkyl, hydroxy, lower alkoxy or lower alkyl. Most preferred are the compounds of formula I wherein R and R ⁶ are hydroxy, lower alkoxy, arylalkyloxy;

R ² and R ⁷ are hydrogen;

R ^J is methyl or amino lower alkyl;

R ⁴ and R ⁵ are bonded via the carbon and nitrogen atoms to form pyrroline, 4-thiapyrrol or 4-methoxypyrrol;

R ¹ is alkyl of 1 to 8 carbon atoms, substituted lower alkyl, the alkyl group having 1 to 5 carbon atoms and the substituent being amino, arylthio or aryloxy, arylalkyl or heteroarylalkyl, the alkyl moiety having 1 to 3 carbon atoms, such as phenethyl, or indolylethyl or substituted arylalkyl (phenyl lower alkyl or naphthyl lower alkyl) and substituted heteroalkyl, the alkyl group of the substituent (s) being / are halogen, dihalogen, amino, aminoalkyl, hydroxy, lower alkoxy or lower alkoxy.

Said preferred compounds also include their pharmaceutically acceptable salts, in particular compounds such as:

N- (1 (3) -carboxy-3-phenylpropyl) -1_-alanyl-1-proline;

N- (1 (S) -ethoxycarbonyl-3-phenylpropyl) -1_-alanyl-1-proline and its salt with mapeic acid;

N- (1 (c) -ethoxycarbonyl-4-methylpentyl) -1_-alanyl-1-proline; N- (1-carboxy-5-aminopentyl) -1_-alanyl-1-proline;

N-α- (1 (3) -carboxy-3-phenylpropyl) -1_-lysyl-L-proline;

N-α- (1 (3) -ethoxycarbonyl-3-phenylpropyl) -E-lysyl-proline;

N-α- [1 (3) -carboxy-3- (3-indolyl) propyl] -1-lysyl-L-proline:

N-α- [1 (3) -carboxy-3- (4-chlorophenyl) propyl] - lysyl-L-proline; N-α- [1 (3) -carboxy-2-phenylthiomethyl] N-lysyl-L-proline;

N - [1- (3) -carboxy-3- (4-chlorophenyl) propyl] <RTI ID = 0.0> lysyl </RTI> -4-methoxyproline;

N-α- [1 (3) -carboxy-5-aminopentyl] <- lysyl-L-proline:

Ethyl N- (1 (3) -ethoxycarbonyl-3-phenylpropyl) -E-alanyl proline hydrochloride;

and others.

The products of formula (I)

R1R3

II II

R — C — C — NH — CH — C l _o II

R 20 and preferred subgroups may be prepared by one or more of the methods described below.

As shown in the examples, the reactive groups which are not included in the condensation, such as amino, carboxy, mercapto and the like. can be protected by methods standard in peptide chemistry before coupling reactions take place and then the protection is removed to obtain the necessary compounds.

Method 1. Reaction Scheme 1 (R ² = H)

R-C-C = O + H ₂ N-CH-CNC-C-R6 NaBHsCN »

R7

II HI

The keto acid (or ester, amide or hydroxamic acid) II is condensed with a dipeptide of formula III in an aqueous solution, optimally near the neutral point, or in a suitable organic solvent (eg CH ₃ CN) in the presence of sodium cyanoborohydride to give I ( R ² = H). Optionally, the intermediate Schiff base, enamine or aminol can be catalytically reduced to obtain product I, for example with hydrogen in the presence of 10% palladium on charcoal or Raney-nickel. The ratio of diastereoisomeric products formed can be altered by the choice of catalyst.

If R and R ⁶ are carboxy protecting groups such as alkoxy or benzyloxy or the like, they can be converted by known methods such as hydrolysis or hydrogenation to a compound of formula I wherein R or R ⁶ are hydroxy groups. This is valid for all subsequent methods carried out under the conditions described.

In one case, the compound of formula II may be fused to amino acid IV

R ³ NaBH ₃ CN O R ¹ R ³

H ₂ NCH-COOH <II --------- ► R — C-CHNHCH — COOH

IV V under the same conditions to obtain amino acid V. By coupling known methods to the amino acid derivative VI, a compound of formula I is obtained.

Known methods include protecting reactive groups during coupling reaction, for example, by N-formyl, N-t-butyloxycarbonyl and N-carbobenzyloxy groups, followed by their removal to obtain a compound of formula I. In addition, the substituent R may be includes removable ester groups such as benzyl, ethyl or t-butyl. Condensing agents for this synthesis are commonly used in the chemistry of peptides such as dicyclohexylcarbodiimide (DCC) or diphenylphosphorylazide (DPPA) or a compound of formula V can be activated by the intermediation of active esters, such as those obtained from 1hydroxybenzothiazole.

R ⁴ R ⁵ DCC

V * HN-C-CO-R ⁶ ------ * I

R ⁷ (VI)

DCC = dicyclohexylcarbodiimide

DPPA = diphenylphosphorylase

Reaction scheme 2

VII

VIII

Amino acid (or ester, amido or hydroxamic acid) VII is fused to ketone VIII under the conditions described for reaction scheme 1 to give a compound of formula I.

Optionally, the synthesis can be carried out in steps by condensing a compound of formula VII with a keto acid of formula IX.

R ³ I VI * O = c — COOH - R ¹ R ³ II —► RC — C — NH — CH — COOH R2 IX X

to obtain amino acid X. By known methods mentioned above, in reaction scheme 1, X may be fused to the amino acid derivative of formula VI to obtain a compound of formula I.

R <R5

I I

X * HN-C-CO — R6

R ⁷

VI

About R1 R3 r4 r5 ο

II I I I I I II R — C — C — NH — CH — C — N — C — C — RS ι ο II 17

R2 O R7

In a case where R ¹ has an α-amino substituent, the carbonyl and amino groups may be suitably protected as a β-lactam group.

Method 2. Reaction pathway 1

R3 O R ⁴ R ⁵

I II I I

H ₂ I4-CH-CNC-CO-R6 * ² I

R7

111

R ¹

I

X-C-COR

I

R2

XI

O R ¹ II I

R ³

I

R ⁴ R ⁵ O

I I II

R-C-C-NH-CH-C-N-C-C

I, II I,

R2 OR ⁷

R ⁶

The dipeptide of formula III is alkylated with the appropriate αhalogen acid (ester or amide) or α-sulfonyloxy acid (ester or amide) of formula XI under alkaline conditions in water or an organic solvent.

X is a chlorine, bromine, iodine or alkylsulfonyloxy or arylsulfonyloxy group.

In one case, the synthesis can be carried out in the following steps:

R3

I h ₂ n-ch — coon

R ¹

I

X-C-COR

I

R2

IV

XI

R1 R ³

I I

RCO-C-NH-CH-COOH

I

R2

X ^ R ⁴ R ⁵

HN-C-COR ⁶

I.

VI

OR ¹ r3 R4 r5 o

II I I I I I II

R-C-C-NH-CH-C-N-C-C-R ⁶

I o II I -, r2 ο R ⁷

X is chlorine, bromine, iodine, alkylsulfonyloxy or arylsulfonyloxy.

The amino acid of formula IV is alkylated with an halogen-containing acid (ester or amide) of formula XI under alkaline conditions to afford compounds of formula X. The latter is condensed by standard methods as indicated in reaction pathway 1 with the amino acid (ester or amide) ) of formula VI to give a compound of formula I.

By reducing the breakdown of a benzyl ester of formula I in which R ⁶ is benzyloxy and R is alkoxy, compounds of formula I are obtained in which R is alkoxy and R ⁶ is hydroxy and when R ⁶ is alkoxy and R is benzyloxy. there are obtained compounds of formula I wherein R is hydroxy and R ⁶ is an alkoxy group.

Reaction scheme 2 R1 I R ³ 0 R ⁴ R ⁵ R-C-C-NH _{? +} I n I I _ II I ² X-CH-CNC-COR ⁶ 0 R2 R ⁷

VI

XI

OR ¹ R3 r4 r5 0

II IIII II _c

RCC-NH-CH-CNCC-R6 l _o II 1 ₇

R 2 OR ⁷ in which X is a chlorine, bromine, iodine, alkylsulfonyloxy or arylsulfonyloxy group.

The amino acid or derivative of formula VII is alkylated with the appropriately substituted α-haloacetyl or α-sulfonyloxyacetyl amino acid of formula XII, preferably R ⁶ other than hydroxyl. under alkaline conditions in water or other solvent to obtain the compounds of formula I.

Optionally, the synthesis can be carried out in steps by condensing the amino acid ester of formula VII with substituted α-halogenated acetic acid or α-sulfonyloxyacetic acid (XIII) to obtain the intermediate of formula X. By known methods described in the reaction 1, X may be coupled to an amino acid of formula VI or a derivative thereof to obtain a compound of formula I. In the methods described, the arylalkyloxy moiety in R or in R ⁶ or both may be used to afford

get an ester or a diester. R-C- R1 I -C-NH, R ³ || I I 0 R2 x-ch-soon VII XHI

R ¹ R ³

I I

RCO-C-NH-CH-COOH

R2

R ³ R ⁴ R ⁵ Q

I I I fl NH-CH-C-N-C-C-R6 II I,

About r7

In one case, the protecting groups may be removed by known methods.

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

In compounds of general formula I, the carbon atoms which are attached to R ¹ , R ³ and R ⁵ may be asymmetric. Therefore, the compounds exist in diastereoisomeric forms or in mixtures thereof. The syntheses described may use racemates, enantiomers or diastereomers as starting materials. When diastereoisomeric products are obtained in the synthesis, the latter can be separated by known methods such as chromatographic and fractional crystallization.

The amino acid portion of the structures is usually preferred

R ¹

II I R — C — C — NH—

I R2

R ³

I —NH — CH — CO and

R ⁴ R ⁵ O

I I I —N — C — C—

I R7 of formula I be L or S configuration, but diastereomers containing D-amino acid have activity dependent on their structures and have advantages in terms of metabolic stability in vivo and can therefore be used in mixture or as pure diastereomeric compounds .

The compounds of the invention form salts with various inorganic or organic acids and 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 and magnesium salts, salts with organic bases e.g. dicyclohexylammonium salts, N-methyl-O-glucamine, salts with amino acids such as arginine, lysine and the like. Organic and inorganic acid salts may also be prepared, e.g. hydrochloric, hydrobromic, sulfuric, phosphoric, methanesulfonic, toluenesulfonic, maleic, fumic, camphorsulfonic acids. Non-toxic physiologically acceptable salts are preferred, although other salts are also useful, for example, in isolating or purifying the product.

The salts may be formed by known methods, by reacting the free acid or the free alkali form of the product with one or more equivalents of the corresponding suitable base or acid in a solvent, such as water, which is then removed in vacuo or by freeze drying or else by exchanging the cations of the existing salt with another cation on a suitable ion exchange resin.

The compounds of the invention inhibit the enzyme, convert angiotensin, and thus block the conversion of the decapeptide angiotensin I to angiotensin II. Angiotensin II is a potent substance that boosts blood pressure. Therefore, a decrease in blood pressure may result from the inhibition of its biosynthesis, especially in animals and humans whose high blood pressure is associated with angiotensin II. The converting enzyme breaks down the vasodressant substance bradykinin. Therefore, inhibitors of the angiotensin converting enzyme may also lower blood pressure by enhancing the effects of bradykinin. Although the relative importance of these and other possible mechanisms is yet to be established, inhibitors of the angiotensin converting enzyme are effective antihypertensive agents in various animal models and are used clinically, for example in mono-patients with renal-vascular, malignant and congenital hypertension . For example, D. W. Cushman et al., Biochemistry 16, 5484 (1977).

Conversion enzyme inhibitors are evaluated with in vitro enzyme inhibition samples. For example, the method described in Y. Piquilloud, A. Reinharz, M. Roth, Biochem is applied. Biophys. Acta, 206, 136 (1970 which measures the hydrolysis of carbobenzyloxyphenylalanylhistidinylivcin. In vivo assessments can be performed, for example, with normal blood pressure rats provoked with angiotensin I by the method of JR Weeks and JA Jones, Proc. Soc. Exp. Biol Med. 104, 646 (1960), and in rat models with high renin values, as described in S. Koletsky et al., Proc. Soc. Exp. Biol. Med., 125, 96 (1967).

Therefore, the compounds of the invention are used in the treatment of high blood pressure. They are also valuable in the treatment of acute and chronic congestive heart failure, in the treatment of secondary hyperaldosteronism, primary and secondary pulmonary hypertension, renal failure and renal vascular hypertension, as well as in the treatment of vascular disorders such as migraine or Raynaud's disease. The use of the compounds of the invention for said and similar disorders is apparent to those skilled in the art.

In the treatment of high blood pressure and the clinical conditions described, the compounds of the invention can be used in compositions such as tablets, capsules or elixirs for oral administration, rectal administration suppositories, sterile solutions or suspensions for parenteral or intramuscular administration, and the like. The compounds of the invention may be administered to animals or humans in need of treatment at doses providing maximum pharmaceutical efficacy. Although the dose depends on the individual needs of the patient depending on the disease, the weight of the patient and the special diets that need to be imposed by other medicines taken at the same time and other factors that are known by those skilled in the art, the dose range is usually about 1 up to 1000 mg per patient per day, either as a single dose or as a single dose. Preferably, the dose is from 2.5 to 250 mg per patient per day, with the most preferred dose being 2.5 to 75 mg per patient per day.

The compounds of the invention may also be administered in combination with other antihypertensives and / or diuretics, for example, the compounds of the invention may be administered in combination with such compounds as amiloride, atenolol, bendroflumethiazide, chlorothalidone, clondin, cryptenaminoacetates, and cryptosineptanidine , guanethidine sulfate., hydrolysin hydrochloride, hydrochlorothiazide, hydroflumethiazide, metolazone, metoprolotartarate, methylcytiazide, methyldopa, methyldopate hydrochloride, minoxidil, pargillin hydrochloride lithiazide, prazosin, resercin, sodium nitroprusside, spironolactone, timolol, trichloromethane, trimethylate, benzthiazide, quinetazone, ticrinaphane, triamterine, acetaminophen, amino mixtures and combinations thereof.

Typically, the individual daily doses for these combinations may be different: from one-fifth of the recommended minimum clinical dose to the maximum doses when given separately.

To clarify these combinations, one of the antihypertensive compounds according to the invention, effectively clinically, at doses of 2.5 250 mg per day can be effectively combined at doses of 0.5 - 250 mg per day with the following compounds at the indicated daily dose: hydrochlorothiazide (15 - 200 mg), chlorothiazide (125 - 2000 mg), ethacric acid (15 - 200 mg), amiloride (5 - 20 mg), furosemide (5 - 80 mg), propanolol (20 - 480 mg), timolol (5 - 60 mg) ) and methyldopa (65 - 2000 mg). The triple drug combination of hydrochlorothiazide (15 - 200 mg) plus amiloride (5 - 20 mg) plus the conversion enzyme inhibitor of the invention (3 - 200 mg) or hydrochlorothiazide (15 - 200 mg) plus timolol (5 - 60 mg) plus inhibitor of the converting enzyme according to the invention (0.5 - 200 mg) are effective compositions for controlling blood pressure in hypertensive patients. Of course, the limits of these doses may be adjusted on a unit basis when it is necessary to determine a separate daily dose that varies with the disease, the weight of the patient, the special diet and other factors.

Typically, these combinations can be formulated into pharmaceutical compositions, which will be discussed below.

About 1 to 100 mg of a compound or mixture of compounds of general formula I or a physiologically acceptable salt is mixed with a physiologically acceptable vehicle, carrier, excipient, binder, preservative, stabilizer, flavoring agent and the like. in a single dose. The amount of active substance in these compositions is such that a suitable dose in the specified range is obtained.

The following are examples of adjuvants that may be included in tablets, capsules, and the like. Binder such as tragacanth, gum arabic, starch or gelatin, an inert substance such as microcrystalline cellulose; disintegrating agent such as starch, gelatinous starch, alginic acid, etc .; a lubricant such as magnesium sgearate; a sweetener such as sucrose, lactose or saccharin; flavoring agent, such as mint, gaultery, cherry oil. When a single dose is in capsule form, it may contain, in addition to the materials mentioned, a liquid carrier such as vegetable or animal oil. Various other materials may be present as coatings or as unit dose modifiers. The tablets can be coated with lacquer, sugar or both. A syrup or elixir may contain the active compound, sucrose, as a sweetener, methyl and propyl parabens, such as a preservative, a coloring agent and a flavoring with a cherry or orange flavor.

Formulations for sterile injectable compositions can be prepared according to conventional pharmaceutical practice by dissolving or suspending the active substance in a solvent such as water for injections, natural vegetable oil such as sesame oil, coconut oil, peanut oil, cottonseed oil and the like. or a synthetic oily carrier such as ethyl oleate or the like. Buffers, preservatives, antioxidants, etc. may be included as required.

The following examples illustrate the invention and particularly preferred embodiments. Preferred diastereomers of these examples are isolated by column chromatography or fractional crystallization.

Example 1. N-hydroxy-2-enylethyl) -1_-apanyl-1-proline.

A mixture of phenylpyruvic acid (753 mg) and lalananyl-b proline (171 mg) in methanol-water was adjusted to pH 6.8 and treated with sodium cyanoborohydride (173 mg) at room temperature until the reaction was complete. The product was adsorbed 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) -1_-alanyl-Lproline. One portion was purified by gel filtration (LH-20) for spectrographic analysis. The NMR spectrum showed a broad singlet at 7.2 complex absorption from 3.0 to 4.6, a multiplet at 2.1 and a pair of doublets at 1.5 ppm (parts per million).

Example 2. N- (1-carboxyethyl) -1_-alanyl-1-proline.

A solution of Llananyl-Lproline (372 mg) and pyruvic acid (881 mg) in water was adjusted to pH 7 and treated with 337 mg of sodium cyanoborohydride at room temperature until the reaction was complete. The product was absorbed on a strong acid ion exchange resin and then eluted with 2% pyridine in water. Sublimation drying afforded 472 mg of N- (1-carboxyethyl) -1_-alanyl-1-proline. NMR and mass spectrogram are consistent with the structure. The NMR spectrum shows multiplets centered at 4.5, 3.7 and 2.2 ppm and a pair of doublets at 1.6 ppm.

Example 3 N- (1-Carboxy-2-cyclohexylethyl) -1_-alanyl-1-proline.

3-Cyclohexyl-2-oxopropionic acid (cyclohexylpyruvic acid) (0.98 g) and 1-alanyl-1-proline (0.22 g) were treated with sodium cyanoborohydride (0.22 g) as described above. A light colored solid of N- (1-carboxy-2-cyclohexylethyl) -1_ alanyl-bproline, 0.31 was obtained. After purification by chromatography, the mass spectrum showed peaks at 340 (molecular ion), 322, 227, 249 and 226. The NMR spectrum showed complex absorption in the range 4.8 to 3.6, as well as peaks (maxima) at 2.2, 1.7 and 1.2 ppm.

Example 4. N- (1-carboxy-5-methylhexyl) -1_-alanyl-bproline.

6-Methyl-2-oxoheptanoic acid (90 g) and L-alanyl • L-proline (0.21 g) with sodium cyanoborohydride (0.21 g) were treated as described above. A white loose mass of N- (1- (1-carboxy-5-methylhexyl) -1-alanyl-1-proline (0.24 g) was obtained. After purification by chromatography, the mass spectrum showed a maximum at 472 (disilyl derivative). The NMR spectrum shows absorption centered at 4.5, 3.65, 2.0, 1.6 and 0.85 ppm.

Example 5. N- (1-Carboxy-3-methylbutyl) -1_-alanyl-1-proline.

Treat 4-methyl-2-oxopentanoic acid (4-methyl-2-leulinic acid) (1.29 g) and 1-alanyl-1-proline (0.32 g) with sodium cyanoborohydride (0.32 g) as described above. A white loose mass of N- (1-carboxy-3-methylbutyl) -1_-alanyl-1-proline (0.40 g) was obtained. One part was purified by chromatography. The mass spectrum showed a maximum at 429 (molecular ion of disilyl derivative minus methyl, 444-15). The NMR spectrum showed resonances centered at 4.4, 3.6, 2.1, 1.6 and 0.95 ppm.

Example 6. N- (1-carboxypropyl) -1_-alanyl-1-proline.

2-Oxobutyric acid (ketobutyric acid) (1.02 g) and 1-alanyl-1-proline (0.37 g) were treated with sodium cyanoborohydride (0.38 g) as described above. Crude N- (1-carboxypropyl) -1-alanyl-Lproline (0.42 g) was obtained. One part was chromatographed for spectral analysis. The mass spectrum shows apparent peaks at 254 (M-18) and 210 (M-62). The NMR spectrum reveals complex absorption from 4.5 to 3.4 multiplet centered at 2.0 and methyl resonances centered at 1.55 and 0.95 ppm.

Example 7 N- (1-Carboxy-2-methylpropyl) -1_-alanyl-1-proline.

A mixture of the sodium salt of 3-methyl-2-ketobutyric acid (1.46 g) and 1-alanyl-1-proline (0.40 g) was treated with sodium cyanoborohydride (0.41 g) as described above. Elution with an ion exchange resin gave crude N- (1-carboxy-2-methylpropyl) -1_-alanyl-1-proline (0.45 g). The product melts at 131 - 142 °. The NMR spectrum showed complex absorption in the range 4.6 to 3.3, a wide multiplet centered at 2.2 and doublets at 1.65 and 1.1 ppm.

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

2-Oxoglutaric acid (1.46 g) i) and Lalananyl-b proline (0.37 g) were treated with sodium cyanoborohydride (0.38 g) as described above. Crude N- (1,3-dicarboxypropyl) -1-alanyl-1-proline (0.47 g) was obtained, m.p. 140-160 ° C. The mass spectrum of the silylated material shows one ion at 517 m / e, equivalent to the molecular ion for the trisilylated derivative minus methyl (532 - 15). The MRI is consistent with the structure. The methyl resonances are centered at 1.4 ppm.

Example 9. N, 1,4-Dicarboxybutyl) -1'-alanyl-1-proline.

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

Example 10. N- (1-carboxy-3-methylbutyl) -alanilyl-isoleucine.

A solution of 1-alanyl-1-isoleucine (150 mg) and sodium salt of 4methyl-2-oxopentanoic acid (564 mg) in water was adjusted to pH 7 and treated with 140 mg of sodium cyanoborohydride at room temperature for several hours. days. The reaction was quenched with a strong acid ion exchange resin, added to a column of the same resin and eluted with 2% pyridine in water. After freeze-drying, 200 mg (84.9%) of a white loose mass of N- (1-carboxy-3-methylbutyl) -1-alanyl-L-isoleucine were obtained. The mass spectrum showed peaks at 460 for the disilyl derivative 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 at 3.7 ppm.

Example 11 N1-carboxy-2-methylbutyl) -1_-alanyl-1-phenylalanine.

A solution of Lalanil-L-phenylalanine (150 mg) and the sodium salt of 4-methyl-2-oxopentanoic acid (483 mg) in water was adjusted to pH 7 and treated with 120 mg of sodium cyanoborohydride at room temperature for several days. The reaction was suppressed with Dautex 50 (H '), the reaction mixture passed through a column of the same resin and eluted with 2% pyridine in water. Freeze-drying afforded 197 mg (88.7%) of a white loose mass of N- (1-carboxy-2methylbutyl) -1_-alanyl-1-phenylalanine. The mass spectrum showed a maximum at 55 for the trisilyl derivative minus methyl (566-15), 479 for the disilyl derivative minus methyl (494-15> and 449 for the trsilyl ¹ d-y mins-COO ~ MS ^: ~ .zz 11 ⁷ ) The NMR spectrum is broad

· Г - - · „* * * '·,, P - η X, -4 £. ,. . - Oh with. . . Kg »I H / · C X *» '· W / w X-> ·. Aw., · .. .W. »Ν · .Xi 4 -4 w. —4 v4 </ h / C ”. Hmm> χΛ \ □ 4-4. C1 4_. 4 ^ .3 h orgp and smgp /.s ⁷ .. si / ί ν! Ntegging is in accordance with ρτιηι / ζτ4 / ηατθ m pz tpiptp € ΈΟΤΜθι_πθΜΜθ nr ZROMATNN TO 3ΠΗφ3Τ ^| - '! ΐ

In a small flask equipped with a pH electrode, mix 1.05 g of L · alanyl-B-lrol! and 1.2 ml of 4 M NaOH. 0.53 g of chloroacetic acid in 1.2 ml of 2 M NaOH was added. The pH is adjusted to - 9. It is heated to 35 °. The pH was maintained at 8-9 for 15 minutes by adding as much sodium as necessary. 0.53 g of urodic acid and sodium hydroxide are added as needed for 15 min. A third portion of 0.53 g of chloroacetic acid is charged, the pH is maintained for 8 - 9 15 min. then for another 15 min at 85 ° C and cooled. The reaction mixture was passed through a Dautex 50 (H ~) column, washed with water and eluted with 2% pyridine in water. The fractions collected showing: · a positive ninchindrin reaction were co-treated with a small eight B 3 vacuum and subjected to Drying by 1 - ⁴ l · ;. p cr D.

4J Cl »w -W. Ca. 4- ''. C »4 · -» 4 t 4- * hs times ¹ var? ex 50 (Na).

in a few milliliters of water and eluted with 0.5 M citric acid and rp was triturated. The necessary

The first product manifests itself first (the ninhydride test. Well separated from the unreacted alanlprol *. The product fraction is concentrated in vacuo to a weight of about 300 mg.

i ν V »> | - 3 · - W. x. FW V- 4- - - · • y - 4-4 -2 · '4z /. 4W, 'A Cl, Cl J. w * \ x / \ 1 I /. I IpC .Ilf iOCl h 3 oxen then went up with 2% ΐιί'Ρι / ί В IN VSD. FMSI ^; c of the product were concentrated in vacuo and freeze-dried. A yield of 417 mg of N-1-carboxymethyl-1-alanyl-1H-propyl was obtained.

BAD ORIGINAL spectra ^'1 j 0 ^{MeC, n} internal vv? NP? Intercalation ^pi 1.58 oom <d J = 6 * ίθ Mr I - Ο f' - 'F ''Ρ L 4' 1. O ¢ 0 ί ,,,,, ηΛ., M JU 'O CO Pl Ο Ο Qp / o-R

68.

Example 13. LH-carboxyethyl) -alanil-L-proline.

Dissolve 45 g of benzyl pyruvic acid and 4.5 g of L-alanine in p-dioxane. with ^and adjusted to 5.5 g of sodium cyanoborohydride then tempeoatura for 6 days NaOH and deyuavya N.4 mixture was stirred cpu hides a lot '' PCC '/ KTO pH was adjusted to>a'^{<CH,: is} ' ^r O' ^, '^' ^{? and} ' ^{: 1} k' / cp '·'' ^{, and!} a

This diffuser is loaded into a column of Dausecko 50 (H ⁴ '). facilitates ^m Venn in ohm - <Dnmlsap with water, washed with an ohm it dioxan - water, then with water The product was eluted with 2% pyridine in water, all fractions were collected and concentrated to dryness in vacuo. This is followed by grinding with water, filtration and washing with water. N o-N-Casoenzoxyethyl-l-alapyl as a mixture.

diastereomic ultrasomal compression of 1 0 g can be obtained from solids containing oe in the mother liquor.

ύΤ6Ό

L-Proline

Π'-ίΜογρ pr ^ pMAM · '! ⁿ | Sl .. 'f'. Lupo ^ five po dn

ML

O.ztvsoen in

HS p f. ι; p ·. the pole is 0 ° for 6 hours. then _ -C 'trempathype

ΗΟι oi bicarzonate. k was chromatographed on a pre-plate silica gel thin layer chromatography plate with ethyl acetate. It tears off the wide strip at rf = 0.5-0.6, eluting with ethyl acetate!

to obtain 212 mg of the mixture, the solvent was distilled off, 1 ι.

by

BAD OP! R L ec'rp -a \ -i-carbobenzoxyethyl-L0 ... v ... ../1 c. I OO it. I t>. ί .0 j ί u Η ν H And τι ί ί 0-4 VX ί 3 CM cv ... i *. . ci π G j. i. genus Add 50 mg of 10% palladium-on-carbon as a catalyst and boil at a pressure of h _? 276 κΝ / μ and room temperature.

F r-ι »1 r» f3 -% - 1 ^ ;. <l? / Λ! \ Ζ sucking through

About Art .t. ks Ci ύ ·. O C * ΓΊ sf kats Ct-

\ 1 - ο τ Κ-Ι! k> I

Γ> Τ

I

r.-i ji · -aci ο Ον rx is ί i io; -L-alanyl-L-proline. The nmr spectrum is comparable c r> r rp.u.iwp 2 and mass spectrometer ^m aryt of the silylated derivative shows

0.75 g of N-d-carbobenzethyl) -L-alanyl in pyridine were dissolved and 7.5 ml of 1 M triethylamine in pyridine were added, cooled and 1.09 g of benzyl hydrochloride added. L-crack ester and 0.678 g of dicyclohexylcarbodiimide. stored at 0 [deg.] C. for 20 h, filtered, and the reaction mixture was concentrated in vacuo. The residue was dissolved in ethyl acetate; the solution was washed with saturated potassium carbonate and then

The organic phase was dried, concentrated in vacuo, and the residue was chromatographed on silica gel with ethyl acetate hexane. to isolate the diastereoisomeric mixture of N- (1-

uh 'n ^A y! _-! 100 P:

, and after processing and sublimation

Su ^ e-n is .3. y ^ ava ^ -10-chaos <o <siietipc-s1nn.) 1- L-προ.'τ.η as the calf's veil ve ^ es. about spgur> Art. 1.65 ρρ ^ ι 'Dog · ;. 1 in - uh, 4pd

V · W V- ·. ^: - I hear Ο - I - And! .0, .. 030%. N- · karsomethoxiethyl; -L-alanyl-L-prslin.

A mixture of a solution of 1.4 g of methyl L-alaninate hydrochloid and 3.1 g of abromoporsionic acid in dioxane was neutralized to pH 9 with sodium hydroxide, heated to 70 ° C and maintained at 30 for 8 minutes by adding 8 to 9 necessary

BAD ORIGINAL cog-ipcTRo warptiee hydroxide After cooling, the mixture is kneaded with ion exchange resin Dautex 50 (H '') _s washed with water and eluted with 2% pyridine in water. The collected product fractions are freeze-dried. . The crude product was purified ^and Res chromatography on an ion exchange column of Dauteks 50 (Na *) in 0 5 M sodium citrate - buffer pH 3.3. The collected product fractions were concentrated to low volume in vacuo and repeated chromatography with Dautex 50 (Ma '). The product fractions were freeze-dried to give pure N- (1-carbomethoxyethyl) alanine.

The intermediate was coupled to the L-proline-t-butyl ester using diphenylphosphorylazide as described in Example 13, then the t-butyl ester was removed by dissolving in trifluoroacetic acid at room temperature for 3 hours, TFA was distilled off and the reaction mixture was purified on a Dautex 50 (H) - 2% pyridine column. as described, and N- (1-carbomethoxyethyl) -1_-alanyl-L-proline is obtained in the form of a mixture of diastereoisomers.

Example 16 N- (1-Methoxycarbonyl-3-methylthiopropyl) -alanil-Lproline.

A solution of pyruvoyl-L -proline (185 mg), L-methionine, methyl esto (600 mg) and sodium cyanoborohydride (200 mg) in 20 ml of methanol was neutralized with dilute methanol solution of sodium hydroxide. After three days at room temperature, the product was absorbed onto a highly acidic ion exchange resin and eluted with 2% pyridine in water to give 80 mg of product. The NMR spectrum showed OCH <at 3.35, 3-CH ₃ at 2.2 δ and CH-CH ₃ at 1.55 and 1.7 δ. The mass spectrogram of the silylated product shows the expected molecular ion at 404 m / e.

Example 17. N- (1 (5) -carboxy-3-methylthiopropyl) -alanil-LAVAP ORIGINAL

A solution of N- (1 (c) -methoxycarbonyl-3-methylthiopropyl) -Ol alanyl · L-proline (127.5 mg; 0 384 mM) in 2 ml of water was treated in a nitrogen atmosphere with 7.82 ml of 0.180 N sodium hydroxide (0 782 mM) ), the kats boil for 2 and a half hours at room temperature. The product was absorbed from the reaction mixture on 30 ml of Dautex 50 (H *) and celuated with 4% pyridine per well to give 73.5 mg, which was further purified through the LH-20 clone to give 55.7 mg of product. The NMR spectrum in D ₂ O shows -S-CH ₃ at 2.1; -CH ₃ at 1.5 and 1.6 δ and the absence of methyl ester. The mass spectrogram of the silylated product shows the expected molecular ion at 462 m / e.

Example 18 N1-methoxycarbonyl-2- (3-indolyl) -ethyl] -alanil-Lproline.

Work as described in Example 16, with the tryptophan methyl ester condensed with pyrovyl-Lproline in the presence of sodium cyanoborohydride to give N- [1-methoxycarbonyl-2- (3-indolyl) ethyl] -alanil-L- proline. The NMR spectrum in CDCI ₃ shows aromatic protons at 6.9 to 7.7, protons adjacent to the aromatic nucleus and adjacent to a nitrogen atom at 2.8 to 3.9: aliphatic methylene protons at 1.4 to 2.7 and alaninemethyl at 1.0 to 1.4. The mass spectrogram of the silylated product shows one ion at 516 m / e and corresponds to the loss of ^ i '· ^{1 g>} n ^, .'G'P 0 ^ · -j' 3?

Example 19 N- [1 (c) -carboxy-2- (3-indolyl) -ethyl] -O2-alanyl-1proline.

It's working. as described in Example 17. the product is hydrolyzed to give the expected dibasic acid. The NMR spectrum at D> Od ₅ Pyr shows 5 aromatic protons at 6.8 to 7.7: protons adjacent to the aromatic nucleus and adjacent to a nitrogen atom at 2.8 to 7.4 and 7 aliphatic protons at 1.0 to 2.2 δ according to the expected structure. The mass spectrogram of the silylated material shows a maximum at 431 m / e, interpreted as a submerged monosilylated ion, which lost one CH ₃ group.

im OHIG1NAL

N 20 <RTI ID = 0.0> N- </RTI> 1-Carboxy-3-phenylpropyl) -1-alanyl-N-4-thiazolidinicarboxylic acid.

-! Ο

<-. γ n ·

Aj 1 Ο kz *

of benzyl ester of

L-tiavsglidin-D-karbsksilpaga acid (2.6 g) were reacted in me ^t nlen * lornd. treated at 0-5 'with triethylamine (1.4 ml). then with cyclohexylcarbodiimide (DCC) (2.3 g) in methylene chloride and the mixture was allowed to stand overnight. After filtration and washing of the filtrate with water and sodium bicarbonate solution, the solvent was distilled off and the product was chromatographed on silica G60 (E. Merck) in ethyl acetate-hexane. The solvent was distilled off from the combined product fractions in vacuo, and the gasoline ester was hydrolyzed in acetonitrile at pH 13.5 (sodium hydroxide) for 1 hour at room temperature. After neutralization to pH 8, hydrochloric acid was washed with ether, concentrated the aqueous layer in vacuo and freeze dried. The tert-butyloxycarbonyl protecting group in 4 M hydrochloride in ethyl acetate was removed, the product was precipitated with ether, filtered and dried to give 1-alanyl-1-thiazolidine-4-carboxylic acid. 0.385 g of condensate are condensed

acid in water using 0.354 g of sodium cyanobohydride by the method described in the example to obtain 0.53 g of a mixture of di- stereoisomers of N- (1-carboxy3-phenylpropyl) -1'-alkanol-1 -4-thiazolidinecarboxylic acid. The NMR spectrum (D ₂ O - NaOD) contains a doublet doublet at 1.2 ppm (3H), Sigglet at 7.1 (5H), wide absorption in the range 1.6 to 2.0 (2H),

range 2.2 to 4.1 and large aq

I »Ύ ₍ , η · 4 k-ν k-h ..; ч \ \ ^ k * .. y .1 1 _g Mk» ίζ fJvH rivtl i “Ci Example 21 m. The mass spectrum of the silylated material shows DCs,> 1 «i k. 1 * * f * ^ * ¹ * rm * h h> chuO. N- (1-carbxy-3-phenylpropyl) - alanyl-L-

pipecolic acid.

By substitution of the thiazolidine carboxylic ester of Example with the hydrochloride of the methyl ester of L-pipecolic acid (1.8 ^d1 is nonyup.p ?. scopine / ^ s & m-η- title eg nc method P: \ · _ ^ Q GrIROr ·? · -Π • 'll .. .. i y,. H. <U. \ _D.

The γ * P spectrum of tCDODi shows a wide multiplet at 1.3 - 1.9 ppm (? N>. One singlet at 7.22 (5H) and a series of multiplets in the range 2.0 - 4.8 ppm. The mass spectrum of the silylated material shows a maximum at m / e = 580 for the disilylated molecular ion.

Example 22 N- (1 Jarboxy-3-phenylpropyl) -1_-alanyl-1_-methylalanine

By replacing the thiazolidine carboxyl ester of Example 20 with the L-N-methylalanine methyl ester hydrochloride (1.5 g), the title compound of the example can be obtained by the method described in Example 20.

Example 23. N- (1-carboxy-1-methylethyl) -1-alanyl-1-proline.

This compound can be obtained by coupling 7.7 g of 2-bromo butyric acid benzyl ester, 2.4 g of lalananyl-bproline tert-butyl ester and 7.0 g of silver oxide in 40 ml of benzene. After heating at reflux for 24 h, an additional 7.7 g of bromester and 7.0 g of silver oxy was added and heating at reflux continued for another 24 h. After cooling, filtering and distilling off the solvent, the product diester was isolated by conventional chromatographic methods. requisite free acid is prepared by removing ^and a tert - butyl ester group in trifluoroacetic acid and the benzyl group by catalytic hydrogenolysis in the established manner.

Example 24. N-Carboxy-1-phenylpropyl) -alanilyl-proline.

A mixture of 4-phenyl 2-acetic acid (1.49 g) and N-alanyl-1-lrolin (0.31 g) in water was adjusted to pH 7.5 with sodium hydroxide solution and treated with sodium cyanoborohydride (0.32 g) overnight. The product was absorbed on a highly acidic ion exchange resin and eluted with 2% pyridine in water to give 0.36 g of crude diastereomeric product, N '(1-carboxy-1-phenylpropyl) -1_-alanyl-bproline. One part is

BAD OFUG'NAL was purified by gel filtration (LH-20) for spectrographic analysis. The NMR spectrum in dimethyl sulfoxide (DMSO) showed aromatic hydrogen 720, a broad singlet at 3.40. wide multiplets at 3.0 to 3.9. 2.67 and 1.94. one doublet at 1 23 and 1 15 The mass spectrum shows a molecular ion at 492 m / e for the dimethylsilylated groups.

Example 25. \ '- (1-krbbsksi-3-feczlgrlgl-lapanyl-lproline.

Grind and sift XAD-2 polystyrene resin (Rohm &lt; i &apos;Co; Determine the 200-400 mesh fraction and load 440 ml in a chromatographic column which is calibrated with 0.1 Μ NH ₄ OH at 95: 5 (v / v water) methanol 350 mg of N- (1-carboxy-3-phenylpropyl) -1_-alanyl-1-proline prepared and purified according to Example 24, dissolved in 10 ml of the same solvent, were charged into the column, eluting with this solvent. The first isomer is swarmed from the column at an interval of 375 400 ml of eluate, the second isomer in the range 440 - 480 ml, the intermediate fractions containing a mixture of isomers. This isomer is freeze-dried to give 130 mg of a white solid After recrystallization from 1 ml of water to pH 3, 94 mg of white needle crystals of m.p. 148-151 ° are obtained, representing the reactive isomer of configuration. S, S, S determined by X-ray analysis. ' ^I7 L · ⁼ * 6 ⁷ 4 · -' it 1 M NO) after drying in vacuo over P.O. ₅ . NMR (DMSO) showed a single doublet for methyl protons at 1.22-·: jj. μ # Ki-ί, vi L and Ci, lz C Cl dxx □ 1 kz i; L><Co ίνί θ, Cz ζζ 4z y P CO 3 CM DciH θ And 122 mg of a white solid are obtained. Recrystallization of 103 mg of 2.5 ml of water with pH to 3 yields 64 mg of white crystals, m.p. 140-145 ° (decomposition). [α] _D = - 101.6 °, (0.1 M HCl) after drying. NMR (DMSO) showed methyl doublet at 1.17 ppm.

EXAMPLE 26 N- (1'C'-ectoxycarbonyl-3-phenylpropyl) -alanilyl-proline.

Ethyl 2- oxo-4-phenylbutyrate (1.03g) and N-alanyl-E-prslin (0.19 g) were dissolved in a solvent containing 1: 1 ethanol-water. Sodium cyanoborohydride solution (0.19 g) in ethanol - water added dropwise at room temperature

BAD ORIGINAL temperature for two hours. When the reaction is complete, the product is absorbed onto a highly acidic ion exchange resin and eluted with 2% π ..: ρ. in water. Product-rich fractions were freeze-dried to give 0.25 g of crude. N-O-ethoxycarbonyl-phenylpropyl) -6-alanyl-6-proline. The mass spectrum shows a molecular ion at 448 m / e for the monosilylated groups. The required isomer is obtained by chromatography.

Example 27. N- (1-Aminocarbonyl-3-phenylpropyl) -6-alanyl-Bproline.

Work as described in Example 26 to condense 2-oxo-4-phenylbutyramide and 6-alanyl-6-proline in the presence of sodium cyanoborohydride to give N- (1aminocarbonyl-3-phenylpropyl) - B-alanyl-B-proline.

Example 28 N- (1-Carboxy-3-phenylpropyl) -6-alanyl-6-tryptophan.

It's working. as described in Example 24, 2-oxo-4-phenylbutyric acid and B-alanyl-striptophan can be condensed in the presence of sodium cyanoborohydride to give N (1-carboxy-3-phenylpropyl) -B-alanyl -B-tryptophan.

Example

2S. N - (1-carboxy-3-phenylphenyl) -B-alanyl-B-4hydroxyproline.

P G on in the presence

B-alanyl-B-4-hydroxproproline is condensed on sodium cyanoborohydride to give N- (1kg carbs.-3-phenylpropyl) -1-B-alanyl-B-4-hydroxyproline NMR spectrum in deuterium methanol shows one doublet , centered at 1.53 ppm (3H), single -: - at 7.13 -5H) and a series of multiplets in the range 2.0 to 4.7 ppm.

Mass, кт The silylanyl material material exhibits the molecular ion of the toysilylated product at m / β = 580.

Example 30. N- (1-carboxy-3-phenylpropyl) -6-serinyl-6-proline.

Work as described in Example 24, as 2-oxo-4-phenylbutyric acid and B-serinyl-B-proline condense in

'' Ή

BAD ORIGINAL accepts sodium cyanoborohydride. to break N- (1-carboxyO ·. -1 -4P fx f I γι '*' X .... r-l I ^ -1 »·> <% r-ι.«..

h / v- • Η '- * ··' · '·; - - D ' ^: H ^; '

MSCC. .iCtlcZ Г0G- <y. '.. ίp6H CCN Pr OCv ιΤ .. 0 3D trisilylated products The NMR spectrum in D ₂ O is consistent with structure.

EXAMPLE 31 N-carboxy-3-phenylpropyl) -phenylalanil-b-rl.

It was done as described in Example 24, as 2-oxo-4-phenylbutyric acid and 1-phenylalanil-1-proline were condensed in the presence of sodium cyanoborohydride to give N- (1-carboxy-3-phenylpropyl) -1_-phenylalanyl -1_-proline.

The mass spectrum shows an ion at 406 m / e for the molecular ion minus water (424-18). The NMR spectrum in D _? O is consistent with the structure.

Example 32 N- (1-Carboxy-3-phenylpropyl) -1_-cysteinyl-1-proline.

Work as described in Example 24, as 2-oxo-4-phenylbutyric acid and L-c-benzylcysteine-L -proline condensed in the presence of sodium cyanoborohydride. The product can then be treated with sodium in liquid ammonia to give N, N-carboxy-3-phenylpropyl) -6-cysteinyl-L proline.

Etc; Measure 33. N- (1-carbxy-3-phenylprop-N -histidinyl-b-leucine.

It's working. as described in Example 24, as 2-oxo-4-phenylbutyric acid and 1-histipinyl-1-leucine were condensed in the presence of sodium cyanoborohydride to give N- (1-carboxy1 G ~ Τ 'K' XfX G ~ '

Ή '-' · ^: 4 ·

XI

Example 34 1 m- (1-Carboxy-3-phenylpropyl) -1_-phenylalanil-1-arginine.

Work as described in Example 24, condensing 2-oxo-4-phenylbutyric acid and b-phenylalanil-yarginine into

.1 the presence of sodium cyanoborohydride. to give N- (1-carboxy3-phenylpropyl) -1_-phenylalanil-1-arginine

Psimes 35. ^ • / -carbocop-3-Phenylpro-plUk-Fe.nilalay.-l-b trip I plowed.

It's working. as described in Example 24, as 2-oxo-4-phenylbutyric acid and B-phenylalanyl-L-tryptophan were condensed in the presence of sodium cyanoborohydride. to give N- (1-carboxy-3-phenylpropyl; -phenylalanil-b-tryptophan.

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

Work as described in Example 24 to condense 2-oxo-4-phenylbutyric acid and 1-phenylalanyl-1-arginine in the presence of sodium cyanoborohydride to give N- (1-carboxy3- (3-indolyl) propyl ) -1_-alanyl-1_-proline. The mass spectrum of the trisilylated material shows a molecular ion at 603 m / e and one M ⁺ - 15 maximum at 588 m / e.

Example 37 N1-carboxy-3-phenylpropyl) -1_-alanyl-3,4-dehydroproline.

A mixture of 3.4-dehydroproline (2.3 g), tert-Boc-1-alanine-Mhidrosuccinimide ester (7.2 g) and sodium carbonate (2.5 g) in a mixture of dscsan-water was co-authored at 0 'overnight.

8V

• Z 'o U T im · G \ O

R \ V / I -¼ w t <>

DdKyVM chu C 4 p νό Cu p c1oЯ n o.

Acute .-., - ca is a t-Boc protecting group with grifluorooietna acid in the usual way and chromatographed on Dauteks-50 (H ') _m elute with 2% pyridine in water as described in Example 2. Dipeptndat is Isolated by freeze drying. This product was condensed with 2-keto-4-phenylbutyric acid as described in Example 24. to give the product in the form of a mixture of diastereomers. The mass spectrum shows a molecular ion at 490 m / e for the distilled product.

BAD ORIGINAL

The diastereomeric mixture (140 mg) obtained above was separated into its components by chromatography on XAD-2 resin. as described in Example 25. The major component (70 mg) first exits the column = - 143 ° (1.3 methanol) The mass spectrum of each component

1'G η ^ ι / υπ «poi» '·· /> LJ nr> t <4 OH rv- \ /' ο οο <Χ-ζ 'U, -X. . -ζ. _. J. . ₍ VX /:> ΗνΠ 'tti ~ ¹ ·, Ч · КУ <Д дтир и м ethyl елил ирaни products.

| Example 36. '- (1-Carboxy-3-phenylpropyl) -1'-alanyl-2methylthiazolidine-4-carboxylic acid.

This compound can be prepared as described in Example 37 by replacing 2.3 g of 3,4-dehydroproline with 2.9 g of 2-methylthiazolidine-4-carboxylic acid.

Example 39. No. (1-Carboxy-3-phenylpropyl) -1_-alanyl-2methylalanine.

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

^Π ρπ ·. ·? Ρ zo capsules filled with solids and containing 50 mg of active ingredient per capsule and auxiliary components, in mg:

per capsule N- (1-carboxy-2-phenylethyl) -C-alanine-1- 50 mg Π ΓΊ 1 · - » ^1; '' j; and the goat 149 mg ^? 'a ^ hezium stearate 1 mg “2g-su ^{r s} (size 1) 200 angles

The N- (1-carboxy-2-phenylethyl) -1_-alanyl-1-proline powder No 60 was ground and then lactose and magnesium stearate were sieved through a tissue sieve No. 60. The combined ingredients were mixed for 10 minutes and then filled into capsules No. 1 of dry gelatin.

Example 41. N- (1-Ethoxycarbonyl-3-phenylpropyl) -1_-alanyl-1-BAD ORIGINAL proline

Solvent / a is e-alanyl-N-proline (7.7 g) and ethyl-2-oxo-4,., Μύ; 1l, l, 4l ml of ethanol was mixed with 64 g of powdered molecular sieves at room temperature for half an hour. Then slowly for 6 hours, a solution of ^{ι ·? 'Ρ ·. ·} ? Ζ cyanoborohydride (2 6 g) in 40 ml of a ^t anog. C a ⁿ d filtration of d * ι ί X 1 d p C d G \> t h I h. 1Ί D, h <I d I vl Ud h C> Λ Η CA С ^ μ d 1 'Ομ DdΚγ JM MV M Cj ί dK C b 0 Μ. The residue was partitioned between chloroform and water. The pH was adjusted to 8 5. The chloroform layer was separated and discarded. The aqueous layer was acidified to pH 2.7 and the product was extracted with chloroform. The chloroform extract was dried over sodium sulphate and concentrated in vacuo to give 10.4 g of mixed high-pressure pyastereomers and HPLC it was found that the desired product was the desired 1 N- (1- / B-ethoxycarbonyl3). -f enyl propyl) -L-ananun-L-nponnH.

NMR spectrum shows aromatic absorption at 7.1 δ and

diastereomeric methyls, such as a multiplet centered at 1.3 δ.

The mass spectrum shows a molecular ion at 376 m / e and a strong maximum at 358 (M * - H _? O).

PGM ί »<19 ..h-hh H_. B1- (1- (S) -ethoxycarbonyl-3-phenylgrogyl) -E-alanyl-E-

ί. rslein maleate.

A solution of Ν-ι 1-ethoxycarbonyl-3-phenylpropyl) -L-alanyl-EDEM ⁿ yn cm ^o is ^{mm, l?} O ^MeOH ^ 3 8 g ^»r 69 ml aietonitoml is toetioa with A 4.25 g • .'e. -eo'-OBd acid in 69 ml of acetonitrile. After stirring for 1 h at room temperature, the solid was filtered off, washed with acetonitrile and air-dried to give 8.4 g of maleate, m.p. 141-145 ° by HPLC with a purity of about 96%. The crude maleate was recrystallized from acetonitrile to give 7.1 g of N- (1 N -ethoxycarbonyl-3-phenylpropyl) N-alanyl-E-proline maleate, m.p. 148-150 ° by HrLC with a purity of about 99%

BAD ORIGINAL

N - (1-ethoxycarbonyl-3-phenylpropyl) -alanil-spoolin

C.sub.3 C.sub.3 g of 1-alanyl-1-proline. 0.206 g of ethyl 2-oxo-4-phenylbut / rat and 1.6 g of molecular sieves in 10 ml of ethanol were hydrogenated at room temperature at 276 kM / m ^? with 0.1 g of 10% palladium on carbon catalyst. After the ingestion was complete, the crude product obtained by filtration and concentration was absorbed on an ion exchange resin (Dautex 50, H ') and eluted with 2% pyridine in water to give 0.224 g bH (1-ethoxycarbonyl- 3phenylpropyl) -1_-alanyl-1-proline. HPLC shows an isomer ratio of 55: 45.

B. N- (1- (S) -ethoxycarbonyl-3-phenylpropyl) N-alanyl-proline maleate.

A mixture of 3 g of L-alanyl-E-proline, 5 g of ethyl 2-oxo-4-phenylbutanoate and 13 g of 3A molecular sieves and 3.6 g of Raney Nickel in 85 ml of ethanol are hydrogenated at 25 ° C and a hydrogen pressure of 276 kM / m 'until hydrogen uptake ceases. The solid was filtered off, washed with 80 ml of ethanol and the filtrates were separated. Analysis by high pressure liquid chromatography (HPLC) showed a ratio of diastereomers 87 13 in favor of the desired product. Ethanol was removed ⁿ cd vacuum to give an oily product which was ~ 5g- ~? r? <π '''N.r,> 4 - p. .o-ι- ^ ιτ- nU ι_Ό Ο LZQP up-rn

J— -. - -. 11 s J I with r-S '** i' - ·. and r. 4, I »IV · v. X. Wt. i. The 11 ν * two-phase mixture was adjusted to 8.6 with 50% sodium hydroxide. The layers were separated and the aqueous phase was extracted with another 2 x 20 ml of ethyl acetate. The aqueous phase was adjusted to pH 4.25 with hydrochloric acid, 12 g of sodium chloride was dissolved in water and the product was extracted with 5 x 12 ml of ethyl acetate. The extracts were collected and dried with sodium sulfate. The desired product (1- (S) -ethoxycarbonyl-3-phenylpropyl) -k-alanyl-cpoline was crystallized as its maleic salt by the addition of 1.86 g of maleic acid. After a scuffle, the sequel to

BAD The salt was filtered, washed with ethyl acetate and dried to give 5.2 g of pure product, m.p. 150-151 ° C.

Example 44 N- (1-Benzyloxycarbonyl-3-phenylpropyl N-alanyl-L-proline.

A solution of L-alanyl-L-proline (167 mg) and benzyl-2-oxo-4-phenylbutyrate (1.20 g) in 5 ml of ethanol was stirred at room temperature with 3 g of Type 4A molecular sieves. Sodium cyanoborohydride (75 mg) was then added portionwise over three hours. The product was purified by absorption in a strong cation exchange resin and eluted with 2% pyridine in water. Passing through a gel filtration column (LH-20) yielded 220 mg of N- (1-benzyloxycarbonyl-3-phenylpropyl) -1_-alanyl-L-proline as a mixture of isomers. After thin layer chromatography on silica gel, eluting with 1 ethyl acetate, 1 n-butanol, 1 water, 1 HOAc (acetic acid) revealed a large major spot with Rf 0 = 71. The isomers were separated using reverse phase HPLC to give N- (1 (S) -benzyloxycarbonyl-3-phenylpropylN'-alanyl-L-proline.

Similarly, reduction of N-acetylaminoethyl-2-oxo-4-phenylbutyrate and L-alanyl-L-proline with sodium cyanoborohydride yields N- [1- (2-acetylamino) ethoxycarbonyl-3-phenylpropyl] alanyl-proline.

Similarly, dimethylaminoethyl 2-oxo-4-phenylbutyrate and Lalanyl-L-proline afforded N- [1- (2-dimethylamino) ethoxycarbonyl-3-phenylpropyl] -1-alanyl-proline.

In the same way, benzyl 2-oxo-4-phenylbutyrate and L-alanyl-Lproline provide N- (1-benzyloxycarbonyl-4-methylpentyl) N-alanyl-L-proline.

The mass spectrum of the silylated material showed peaks at 510 (M ~ -15), 464, 420, 419, 385 and 296 m / e (base maximum).

The NMR spectrum is consistent with the structure.

Example 45 N- (1-Butoxycarbonyl-3-phenylpropyl) -E-alanylL-proline.

A solution of 3 ° N-benzyloxycarbonyl- ^4- methylpentyl) -alanilyl-bproline (452 mg) in 5 ml of benzene was hydrogenated over 150 mg of 10% palladium on charcoal to remove the nitrogen protecting group. After filtration and evaporation of the solvent, the Lalanil-L-proline 3 ° butyl ester was dissolved in 8 ml of tetrahydrofuran and treated with 1.41 g of butyl 2-oxo-4-phenylbutyrate and 3 g of powdered molecular sieves. Sodium cyanoborohydride (150) was added portionwise over several hours, stirring the mixture overnight at room temperature. After filtration and concentration in vacuo, the residue was treated with 25 ml of trifluoroacetic acid for 2 hours at room temperature. After removal of the acid, the product was purified by absorption on an ion exchange resin and by gel filtration (LH-2O). Concentration and drying of the product fractions afforded 182 mg of N- (1-butoxycarbonyl-3-phenylpropyl) -Lanylanyl-L-proline as a mixture of isomers. After thin layer chromatography (silica gel, 1 ethyl acetate, 1 butanol, 1 water, 1 acetic acid) two spots with Rf 0.67 and 0.72 are shown. The mass spectrum shows peaks at 548 (disilylated molecular ion) and 4 ⁷ 6 (monosilylated molecular ion). The isomers were separated by reverse phase HPLC to give N- (1- (S) butoxycarbonyl-3-phenylpropyl) -1-alanyl-L-proline.

Example 46 N- (1-Ethoxycarbonyl-3-phenylpropyl) -1-alanylL-proline, ethyl ester.

A solution containing 0.63 g of N- (1-carboxy-3-phenylpropyl) -1 alanyl-L-proline in 9.7 ml of ethanol was saturated with HCl gas at 0 ° After standing overnight at room temperature, HCI and ethanol were removed in vacuo, to obtain a light yellow oily liquid which was purified by gel filtration (column LH-20). Obtained from ethyl ester. N- (1-ethoxycarbonyl-3-phenylpropyl) -L alanyl L-proline is 0 39 g, one spot on thin layer chromatography. The NMR spectrum shows two ethyl groups for the aromatic ring. The mass spectrum shows a molecular ion at 404 m / e.

Example 46. N- (1-Carboxy-3-phenylpropyl) -E-alanyl-trans-4methoxy-L-proline.

Methyl trans-4-methoxy-prolineate hydrochloride was prepared by the method of E. Adams et al., J. Biol.Chem. 208, 573 (1054) by esterification with methanol hydrogen chloride in the standard manner. BocL-alanine in methylene chloride was combined with dicyclohexylcarbodiimide as described above, and the intermediate Boc-L-Ala-L-methoxy Pro-OMe was purified by chromatography on silica gel and eluting with ethyl acetate: hexane in a ratio of 1: 1 . The ester is hydrolyzed with sodium hydroxide in acetonitrile-water, adjusted to pH 7.5, dried by sublimation, and the amine is liberated in 4M hydrogen chloride in ethyl acetate in the usual way. 0.54 g of this L-alanyltrans-D-methoxy-L-proline was condensed with 2.0 g of 2-oxo-4-phenylbutyric acid in 6 ml of water using 0.43 g of sodium cyanoborohydride as described in Example 24. After isolation according to Example 24 yields 0.92 g of a mixture of diastereoisomers of. N-Cl-carboxy-3-phenylpropyl) E-alanyl-trans-4-methoxy-L-proline. The NMR spectrum in D ₂ O shows a split doublet centered at 1.58 ppm (3H), singlet at 3.37 (3H) and 7.35 ppm (5H), complex absorption in the range 1.9 - 3.5 and wide multiplet at 4.0 - 4.6 ppm. The mass spectrum shows protruding peaks at pte 360 (M-18) and 256 (M-122).

Example 48 N- (1-Benzyloxycarbonyl-3-phenylpropyl) N-alanyltrans-4-methoxy-L-proline.

By binding L-alanyl-4-methoxyproline. obtained according to Example 47, with 2-oxo-4-phenylbutyrate in ethanol in the presence of sodium cyanoborohydride according to the method described in Example 44, a mixture of diastereoisomers of N- (1-benzyloxycarbonyl-3-phenylpropyl) -1-alanyl-trans can be obtained -4-methoxy-L-proline. The isomers can then be separated by reverse phase HPLC to give N- (1 H -benzyloxycarbonyl-3-phenylpropyl) N-alanyl-trans 4-methoxy-L-proline.

Example 49 N- (1-Benzylaminocarbonyl-3-phenylpropyl) -1-alanylL-proline.

2-Oxo-4-phenylbutyric acid benzylamide was obtained by dissolving 3.0 g of this acid, 2.4 ml of benzylamine and 4.7 ml of diphenylphosphoyl azide in 60 ml of cold dimethylformamide and adding dropwise 2.6 ml of triethylamine in dimethylformamide (DMF). maintained the temperature at about -10 ° C for 2.5 hours and overnight at room temperature. The dimethylformamide was distilled off 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 1: 4. Evaporate the solvent from the product fractions to give 2.2 g of crystalline N-benzyl-2-oxo-4-phenylbutyramide. Bind 1.26 g of it with 0.19 g of L-Ala-L-Pro. using 0.125 g of sodium cyanoborohydride in ethanol as described in Example 44. The crude product was purified by gel filtration (LH 2 O). to obtain a mixture of diastereoisomers of N- (1-benzylaminocarbonyl-3-phenylpropyl) -k-alanyl-L-proline. The NMR spectrum (COCl ₃ ) showed one doublet at 1.1 ppm (H). a pair of singlets at 7.3 (10H) and complex absorption at 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 clearly expressed maxima at m / e = 509 (monosilyl derivative) and 581 (disilyl derivative).

Example 50 N- (1-Carboxy-3-phenylpropyl) -1-alanyl-N-methylphenylalanine.

By replacing the thiazolidine carboxyl ester of Example 20 with N-methyl-L-phenylalanine methyl ester, Lalananyl-1-methyl-1-phenylalanine is obtained. Condense 0.85 g of it with 3.02 g of 2-oxo-4-phenylbutyric acid using 0.64 g of sodium cyanoborohydride in water as described. The mixture was acidified to pH 1 5 and extracted into ether. The ether was distilled off and the residue was dissolved in 70% methanol-water and chromatographed on a Dautex 50 (H ') column. prepared with this solvent. It was eluted with a solution of 3% pyridine in the same solvent. The product fractions were collected, concentrated, dried by sublimation and purified on LH-20 in methanol. to give 0.54 g of a mixture of isomers of. N- (1carboxy-3-phenylpropyl) -1_-alanyl-N-methyl-phenylalanine. The NMR spectrum (D ₂ O, NaOD) showed one doublet centered at 1.18 ppm (3H), two overlapping singlets at 7.3 (10H), single at 2.95 (3H) and wide multiple absorption in the ranges 1.4 to 2.1 and 2.3 to 4.4 . The mass spectrum of the silylated product indicates the molecular ion of the desilylated product at m / e = 556.

Example 51 N- (1-Ethoxycarbonyl-3-phenylpropyl) -1-alanyl-1-prolineamide.

L-alanyl-L prolineamide is obtained by coupling tert, Boc-L-alanine with L-prolineamide by known methods using 4: 1 methylene chloride: dimethylformamide dicyclohexylcarbodiimide. The intermediate tert-Boc-L-Ala-LΡγο-Ν--Κ was purified by chromatography on LH-20 in methanol, then the tert-Boc protecting group in 4 M HC was removed! in ethyl acetate. Bind 0.5 g of product L-Aia-L-Pro-NH ₂ .HCl in 10 ml of absolute ethanol, neutralized with one equivalent of triethylamine, with 2.4 g of ethyl 2-oxy-4-phenyl-butyrate using molecular sieves and 0.30 g sodium cyanoborohydride as described in Example 41. In this example, the product is contained in a chloroform extract at pH 8.5; concentrated in vacuo, dissolved in 50% ethanol - water and chromatographed on Dauteks 50 (H ^g), prepared in 50% ethanol-water, eluted with 2% pyridine in this solvent, the product fractions were collected and was further purified by chromatography on LH-20 in methanol. The solvent was distilled off in vacuo to give 0.40 g of N- (1BAD O. 'ethoxycarbonyl-3-phenylpropyl) -1_-alanyl-1-prolineamide in the form of diastereoisomers NMR (CDCIJ shows one triplet overlapping doublet). 1.5 ppm (6H) series of five multiplets in the range 1.5 - 4.7 ppm (Ί5Η) and singlet at 7.17 ppm (5H) The mass spectrum of the silylated material shows clearly expressed maxima at m / e = 477 (monosyllable derivative) and 519 (disilyl derivative). derivative).

Example 52 N- 1- (1-Carboxy-3-phenylpropyl) -1_-alanyl-1-prolineamide.

The 1-alanyl-L-proline amide, prepared as described in Example 51, was coupled to 2-oxo-4-phenylbutyric acid using cyanoborohydride in 50% ethanol-water by the method described in example 2. After elution with the ion exchange resin, it is concentrated in vacuo to a small volume, washed with water and freeze-dried to give N- (1-carboxy-3-phenylpropyl) L'anyl-L-proline-amide as a mixture of diastereoisomers.

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

Example 53. N- (1- (S) -hydroxyaminocarbonyl-3-phenylpropyl) - [a] apanyl-bproline.

To a cold solution of N- (1-ethoxycarbonyl-3-phenylpropyl) -1-alanyl - [Proline-maleate (0.19 g), obtained as described in Example 42, in 1 ml of ethanol. 0.85 g of potassium hydroxide in 0.57 ml of ethanol is added. Subsequently, a suspension of 0.07 g of hydroxylamine hydrochloride in 0.9 ml of ethanol containing 0.060 g of potassium hydroxide is added dropwise. It is kept in an ice bath and then at room temperature overnight. The top layer was decanted, diluted with 10 ml of water, adjusted to pH 2.5 with hydrochloric acid and washed with chloroform. The aqueous layer was neutralized and freeze-dried and purified by chromatography on XAD-2 resin in a concentration gradient of 0.1 M ammonium hydroxymethanol to give N- (1- / S / hydroxyaminocarbonyl-3-phenylpropyl) -1_-alanyl - Proline. The massspecter friction of the silylated material shows one ion at m / e = 579 for the trisilylated derivative and the NMR is consistent with the structure / rt

Example 54. N- (1-Carboxy-3-methylbutyl) -alanil-B-tryptophan

A solution of the sodium salt of 4-methyl-2-oxopentanoic acid (414 mg) and B-alanyl-B-tryptophan (150 mg) in water was adjusted to pH 7 with sodium hydroxide and treated with sodium cyanoborohydride (103 mg) at room temperature for several days. The product was absorbed on a highly acidic ion exchange resin and eluted with 2% pyridine in water. Product-rich fractions were subjected to freeze-drying to give 189 mg of white powder. The mass spectrum shows a molecular ion at 389 m / e and peaks at 187 m / e and 157 m / e for the fragments shown separately:

sleep-sleep ₃

I

CH ²

but ₂ s-ss-nh-ss

NMR spectrum in D _? O is consistent with the structure.

Example 55. N- (1-Carboxy-3-methylbutyl) -6-histidyl-6-leucine.

It was done as described in Example 54, as 4-methyl-2-oxopentanoic acid and 6-histidyl-6-leucine were condensed in the presence of sodium cyanoborohydride to give N- (1-carboxy3-methylbutyl) -6-histidyl- B-leucine. In this example, the product was eluted from the ion exchange resin with 10% ammonia. The mass spectrum is in accordance with the structure / rat.

Example 56. N- (1-Carboxy-3-methylbugyl) -6-phenylalanil-barginine.

Work as described in Example 54, with 4-methyl-2-oxopentanoic acid and 6-phenylalanyl-6-arginine condensed into

BAD ORIGINAL presence of sodium cyanoborohydride to give N- (1-carboxy-3-methylbutyl) -1_-phenylalanil-1-arginine. The product was eluted from iopoform resin with 10% ammonia. The NMR spectrum in D _? 0 shows one split at ⁷ 2 (5H), a wide triplet centered at 3.0 (6H), a wide multiplet at 1.3 (6H) and one wide doublet at 0.9 (6H).

Example 57

A. N- (1-Carboxy-3-phenylpropyl) -1_-lysyl-Lproline.

Work as described in Example 54 as 2-oxo-4-phenylbutyric acid and tert-Boc-lysyl-L -proline condensed in the presence of sodium cyanoborohydride. Essentially all the third Boc protecting groups are cleaved when the product is absorbed on a highly acidic ion exchange resin. The crude N- (1-carboxy-3-phenylpropyl) -Lysyl-1-proline was eluted from the resin with 10% ammonia, dried by sublimation and purified by gel filtration chromatography (LH-20). A barely noticeable maximum for tert-Boc protons in the NMR spectrum disappears when the product is treated with 4 N ethyl acetate in hydrogen chloride gas. The NMR spectrum of the resulting hydrochloric acid salt of the product is consistent with the structure. The mass spectrum shows a molecular ion at 693 m / e for the tetrasilylated products. Chromatography on XAD-2 resin using 3.5% acetonitrile in 0.1 molar ammonium hydroxide gave N- (1-carboxy-3-phenylpropyl) -1_-lysyl-1-proline.

B. 1 1-a- (1 / in / -carboxy-3-phenylpropyl) -1_-lysyl-1_-proline.

Work as described in Example 54 as 2-oxo-4-phenylbutyric acid and N-tert-Boc-lysyl-E-proline are condensed in the presence of sodium cyanoborohydride. The product was absorbed on a highly acidic ion exchange resin and eluted with 2% pyridine in water. The product-rich fractions were distilled off and treated with 4 N HCl in ethyl acetate to remove tert-Boc protecting groups. The resulting hydrochloride salt is converted to the free base by absorption on a highly acidic ion exchange resin and eluting with 2

BAD OFliGiNAL% pyridine in water. Sublimation drying of product-rich fractions gave Ν-α-ί 1 (in) -carboxy-3-phenylpropyl) -1-lysyl-µ-proline as a white loose mass. The NMR spectrum is consistent with the structure. The mass spectrum shows a molecular ion at 549 for the distillated products. Chromatography afforded the required isomer.

Example 58 N- (1-Carboxy-3-phenylpropyl) -1- 3 -fluoroalanil-1-proline.

To a solution of L-3-fluoroalanine (420 mg) in 4 ml of acetone-water (1: 1) was added triethylamine (589 mg) and 2-tert-butoxycarbonyloxyimino-2-phenylacetonitrile (1.060). The mixture was stirred for 2.5 hours, cold aqueous potassium bicarbonate solution (5%) was added and the mixture was extracted with ethyl acetate. The aqueous phase was acidified with cold 1Ν hydrochloric acid and extracted with ethyl acetate. The 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) m.p. 91 - 93 ⁰ .

To a stirred solution of the resultant compound (800 mg) and prolinebenzyl ester (1.5 g) in methylene chloride (8 ml) at 0 ° was added dicyclohexylcarbodiimide (845 mg) in methylene chloride (6 ml) and stirred with C psyde at 0 ° for 2 hours and at 20 ° - 18 hours. The mixture was filtered, the precipitate was washed with methylene chloride and the combined filtrates and the washings were extracted with cold 1 N hydrochloric acid, cold 5% aqueous potassium bicarbonate and saturated aqueous sodium chloride, dried over sodium sulfate and concentrated. until dry. After silica gel column chromatography eluting with 6% acetone in chloroform, the pure protective dipeptide was obtained.

The tert-Boc group was removed by treatment with 4 N hydrogen chloride in ethyl acetate (8 ml) at 0 ° for 1 hour. About 20 ml of ether was added and the hydrochloride of the L3-fluoroalanil-L-proline gasoline ester (450 mg) was precipitated, m.p. 158-161 ° C, which was separated

BAD ORIGINAL by flutonation After hydrogenation in 6 ml of water and 2 ml of ethanol over 60 mg 10% palladium on carbon at a pressure of ¹ atmosphere and 20 ° C in

dry 330 mg of 1_-3-fluoroalanil-1-hydrochloride hydrochloride was obtained

The mass spectrum shows a molecular ion at 348 m / e for the trimethylsigilated product.

To a mixture of 4-phenyl-2-oxobutyric acid (375 mg) and L-3fluoroalanil-L-proline hydrochloride (100 mg) in 3 ml of water (pH adjusted to 7 with sodium hydroxide) was added sodium cyanoborohydride (80 G; The mixture was stirred for 20 hours and treated as described in Example 24. The mass spectrum of the LH-20 purified product showed a molecular ion at 510 m / e for dimethylsilyl wash materials Thin layer chromatography on silica gel showed a single spot Rf = 0.7 - system 1 : 1: 1: 1 ethyl acetate: acetic acid: n-butanol:

water.

The diastereoisomers were separated on an XAD-2 resin as described in Example 25.

N- (1-ethoxycarbonyl-3-phenylpropyl) -1_-3-fluoroalanil-L-proline was obtained as described in Example 26.

Example 53 N- (1-Ethoxycarbonyl-1-yl-3-phenylpropyl) -1_-alanyl-1-3,4-dehydropropoline.

'- • Alden-O-3 4-dehydroproline obtained according to Example 37 is converted by the procedure described in Example 26 into' - (1-ethoxycarbyl, 3-phenylpropyl-1-alanyl-1-3 _: 4-dehydropropoline as one two component diastereomeric mixture, thin layer chromatography - silica gel plate Rf 0 82 (basic component) and Rf 0.79 (minor), two manifestations in the 4.1: 1 system - butanol: water: acetic acid. materials.

Example 60 N- [1 (S) -Methoxycarbonyl-2- (1H-imidazol-4-yl) -ethyl] -Alananyl-L-proline.

BAD ORIGINAL

_...... J

Work as described in Example 16, such as methyl

N - ['- metho x and carbon 2 and * on /. see

(1H-imidazol-4-yl, i-ethyl] - C3-alapyl-E-proline.

The NMR spectrum in D ₂ O shows imidazole protons at 8.6 and 7.3;

melts adjacent to the indasla. and protons of methyl ester at 3.7 and alanyl methyl: 1.1 to 1.33.

Example 61 N- [1 (S) -Cap6oxy-2- (1H-imidazol-4-yl) -ethyl] DL-alanyl-E-proline.

Work as described in Example 17, hydrolyzing the product of Example 60 to give the expected dibasic acid. NMR in D ₂ O shows imidazole protons at 7.2 and 8.5; alanine methyl

at 1.25 δ. Example 62. N- [1 (c) -ethoxycarbnyl-5-aminopentyl] - DL-alanyl-E-proline.

A solution of c-benzyloxycarbonyl-lysine ethyl ester, hydrochloride (2.94 g) in water (10 ml) was basified with 15 ml of saturated aqueous potassium bicarbonate solution and extracted with methylene chloride. The extract was dried over magnesium sulfate and concentrated to dryness.

The residue is ε-benzyloxycarbonyl-E-lysine ethyl ester. dissolves

powder molecular sieves No. 4A (1.0 g). The mixture was stirred at room temperature for 4 hours. After 2 hours, sodium cyanoborohydride (630 mg) was added in 1 ml of οπθιπβ acid allowed to stand overnight. the swelling continued, then filtered, concentrated to dryness and the residue partitioned between water (0 mL) and methylene chloride (15 mL). The aqueous phase was absorbed on a highly acidic ion exchange resin and eluted with 4% pyridine in water to give 470 mg of N- [1 (c) -ethoxycarbonyl-5-benzyloxycarbonylaminopentyl] -O1-alanyl-1-proline. The protecting group is removed by hydrogenation in ethanol - Boda 1: 1 over 10% palladium

on coal at 276 kN / m '. The mixture was filtered and the filtrate evaporated to dryness. The residue in methanol is chromatographed on a hH-20 column to give the desired N- [1 (3) -ethoxycarbonyl-5-aminopentyl] □-alanyl-c-proline. The mass spectrum of the silylated material shows a maximum at about 59 ° to iris. ' , the splinted derivative and the maximum in the derivative. NMR in D? O confirms the structure.

Example 63 N- [1 (3) -carboxy-5-aminopentyl] -1-alanyl-1-proline.

N- [1 (S) -ethoxycarbonyl-5-benzyloxycarbonyl-aminopentyl] Lalananyl-bproline. prepared as described in Example 62, was treated with 0.1 M sodium hydroxide at room temperature overnight. The product was absorbed on a highly acidic ion-exchange resin and eluted with 4% pyridine per well to give N- [1 (c) -carboxy-5-benzyloxycarbonylaminopentyl] -OR-alanyl-E-proline. single stain by thin layer chromatography (Rf 0.4 - butanol: water: pyridine: acetic acid 10: 4: 3: 1). As described in Example 62, the protecting group was removed by hydrogenation to give. N- [1 (S) carboxy-5-aminopentyl] -6-alanyl-OE-proline. The mass spectrum of the trislined product is consistent with the structure, it shows

Example 64. N- [1-Carboxy-6-aminohexyl-1-alanyl-b-proline.

Benzyl 2-oxo-7-phthalimidoheptanoate. obtained by alkylation of benzyl 1 3-dithane-2-carboxylate with 5-phthalimidopentyl bromide and subsequent oxidation to ketone with N-bromosuccinimide, is condensed with B-alanyl-B-proline in the presence of excess NaBH ₃ CN. The condensation product of 1- {1- [1-benzyloxycarbonyl-6-phthalimidohexyl] - n-alanyl-L · proline (390 mg) was hydrogenated in 25 ml of 50% aqueous ethanol at 276 kN / m ² over 10% palladium on charcoal. Removal of solvent and catalyst is obtained. N-8 [1-carboxy-6-phthalimidohexyl] -1'-alanyl-1-proline (320 m 2 with the expected spectral and chromatographic properties. One hour 'of the intermediate obtained (152 mg · in 2 ml of ethanol was heated refluxed with hydrazine (32 mg) <.5 hours, the phthalhydrazine was removed by filtration, the ethanol was removed in vacuo, and the residue was aspirated on a highly acidic ion exchange resin After elution with 2% aqueous pyridine and freeze-drying N was obtained. fi-carboxy-6-aminohexyl] -1-alanyl-1-proline (58 mg) The mass spectrum shows a maximum at 311 per molecule borate ion minus water (329-18).

Example 65. N-fl-Benzyloxycarbonyl-6-aminohexyl] -L-alanyl-L-proline.

By performing hydrazinolysis as described in Example 64 on N-t-benzyloxycarbonyl-6-phthalimidohexyl] -1-alanyl-1-proline, a mixture can be isolated from which N- [1-benzyloxycarbonyl-6-phthalimidohexyl] - L-alanyl-L-proline. After thin layer chromatography on silica gel, eluting with 50 butanol: 11 acetic acid: 30 water, a spot is obtained, Rf 0.32.

Example 66 N- [1-carboxy-2-phenoxyethyl] -L-alanyl-L-proline.

Suspension of phenoxypyrazole acid (1.8 g), obtained by xenesis of ethylphenoxyacetate with diethyloxalate, followed by acid-catalyzed hydrolysis and decarboxylation and L-aaa-yl sodium hydroxide. The mixture was treated with NabH ₃ CN (0.18 g) and allowed to stand

'pn constant stirring ^and a 5 third day an additional ketoacid 10.9>! and sodium cyanoborohydride (0.18 g). The product was absorbed on a highly acidic ion exchange soma and eluted with 2% pyridine in water to give, after freeze-drying, 0.5 g of N- [1-carboxy-2-phenoxyethyl] - L-alanyl-L-proline. The NMR spectrum is consistent with

,? _{The τ3} l / zse-e / .ter shows one maximum at 479 for h / · / ;. 'c ·' mn-, ·? m-zol; 494 15).

with / '.' cp ο ·. _L ι-e'Zle »iKapucr.i'l-2-fenS \ s.ethyl] - L-alag-yl-lps-olin

By reacting the ethylphenoxypyruvate obtained from the acid by acid-catalyzed esterification and L-alanyl-Lproline with NaSH2CN is an ethanolic solution and isolation of the product as described in Example 6b, N- [1-ethoxycarbonyl-2-phenoxyethyl] can be obtained - ! _- and ⁿ anil- L-

Example 68. N [1-Carboxy-2-phenylthioethyl] - L-alanyl-L-proline.

A mixture of phenylthiopyruvic acid (1.96 g), obtained by condensation of ethylphenylthioacetate with diethyloxylate, followed by acid-catalyzed hydrolysis and decarboxylation, and L-alanyl-Lproline (0.37 g) in 10 ml of water, was adjusted to pH 7.0 with dilution to pH 7.0 with dilution to pH 7.0. was treated with NaBH ₃ CN (0.18 g) in 2 ml of water. After stirring overnight at room temperature the product is absorbed var ^y y whitefish ^ o acidic ion exchange resin and eluted with 2% pyridine in water to yield 0.36g of N- [1-carboxy-2feniltioetil] - L- alanyl-L-proline. The NMR and mass spectrum show the desired structure. One maximum at 348 indicates the molecular ion »-V ·» proline.

Mr. ·>! l · I mutual :, wasp

8. may have the product as described to give N- [1-ethoxycarbonyl-2-phenylthioethyl] z * in example

L-alanyl70. N- [1-ethoxycarbonyl-3-p-chlorophenylpropyl] alanyl-L-proline.

L50 Ethyl 4-p-chlorophenyl-2-oxo-butyl solution. acid through esterified? with ethanol in CC ·,. and _ al:

obtained from rt 'r-l ΓΊ I | I I | m Ci 3 7Ί I m I L / C Gbp Kci I i jj ». room ”*? until the reaction is complete, the eganolt can then be removed in vacuo and the product absorbed on a highly absorbed ion exchange resin. Eluting with 2% pyridine so-da MO.r.e to give N-fl-ethox / zharsg-nl-3 p-chloro-phenyl] -pyrrol] ·, Α

Tooth

L71. DG [1-Ethoxycarbonyl-2- (3-indsyl) ethyl] - L-alanyl

Lproline.

as in Example 26. as the indole-3-acid ethyl ester is condensed with L-alanyl-L-proline in 1: 1 spectrum shows a pyrogen-free solution of ethanol: water by sodium cyanoborohydride. Isolation of Dutex 50 as described above provides a mixture of isomers of N- [1-ethoxycarbonyl-2- (3-indolyl) ethyl] -L-alanyl-L-proline. NMR absorption at 7 0 δ * 5H). 4 2δ (5H). 3.4 δ (2H), 1.9 δ (6H) and 1.2 δ (6H).

Example 72

N- [1-ethoxycarbonyl-2-p-aminosmethylphenylethyl] - Lalanyl-L-proline.

- 4 schpenpleropanoate obtained nd p-oiaobenz ·. with ethyl 1-10-ditpan-ho carbecenlate and subsequent oxidative hydrolysis as described by Elie: and ^u anmann. J. Org. Chem., 37, 505 (1972)) with L-alanyl-L-proline and the product 'was purified by the method described in Example V. The resultant isomer of the - (1-ethoxycarbonyl-2-hydroxypropyl) L-alanyl-L-pooline in ethanol solution, containing hydrogen chloride and palladium-on-carbon catalyst. The solvent and excess HCl were distilled off in vacuo, washed with ethanol and concentrated to dryness to give hydrochlorides. of the diastereoisomers of the desired compound.

η

uh. .and ORIGINAL ^ ^π ρΐ ". · θρ 73 N- [1-carboxy-2-p-aminomethylphenylethyl] - L-apanyl {_-psylline is recovered from the N- [I-ethoxylcarbonyl-3-p-thiophenylethane-alanyl] - Proline obtained in Example 72 with one equivalent of sodium hydroxide in a mixture of methanol and water as a solvent at room temperature for one ns. The solvents were removed in vacuo to afford the sodium salts of a mixture of N- [1-carboxy-2-cyanophenylethyl] - L-alanyl-L-proline. Hydrogenate this mixture in ethanol solution of hydrogen chloride and serogroup as described in Example 72 to obtain the hydrochlorides of a mixture of measurements of the desired compound.

Example 74 N- [1-Ethoxycarbonyl-2 (S) -amino-3-phenylpropyl] -O. Lalanyl-L-proline.

To a mixture of N-phthaloyl-1-2-amino-3-phenylpropionaldehyde (Peterson et al., J. Am.Chem.Soc. 79. 1389 (1957)), (2.18 g) and potassium metabisulphite (0.87 g) in water: methanol 1: 1 sodium cyanide (0.55 g) was added with vigorous stirring. It was stirred for 90 min, 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-phthaloyl-3-amylC-3-methyl-N-hydroxythiothyronitrile.

of these prduK (in o-ssvsdsp ι 1 Q L

stay for two days at room temperature. Removed: conc. hydrochloric acid solvent, dissolve the residue in dioxane: (1: 1). heated to 7C 'and maintained at ~ ~ €

· "V" ~ / '• y ·' ·. η POitJ p O O t S O

4Z _? I - \ - · - · w G “*! '-Z J' X /, I X- * XX 'Vi »I C · 4- X. I X> —X

s_z / i 1 i k '/. ». L cl C xy And C Θ i D C D aV 4 -, .- xz. . - C - .X.X. . X- and 14-C Civi s. . ,. N, 4-, v; ., 0 custom! · whose mother. to give N, e>: 38) * 2-amino-4-phenyl-o-

phthalosilamino butanoic acid. It dissolves the acid in anhydrous

ethanol, anhydrous hydrogen chloride is passed until saturated and saturated

left at room temperature for 16 hours. The solvent is removed

ο. ORIGINAL j

vacuum to obtain the hydrochloop of ethanol ^< ? este ^m a: z ethyl 2-am-. ·: -ο-όβπ;:;: - 3-0talylamines butanoate with Γ. · ρ _ζ 5O -. 't sodium-cyanoborohydride as described in Example 16. to give N- [1-ethoxycarbonyl-3-phenyl-2-phthaloylaminopropyl] -0, L-alanyl-L-proline in the form of isomeric and D rsdukta s ethanol shaped condenser and with one equivalent of hydrazine for 1.5 h. cooled and stoiltoioa phthalhydrazide, and isolate the desired ⁿ ooduk ^m ^ of ^ s deriving or obtained mixture ^and p? h chromatographic methods to obtain N- [1- ethoxycarbonyl-2 (c) amino-3-phenylpropyl] -O. L-alanyl-L-proline. The 300 Mhz NMR spectrum has a maximum at 7.3 δ (5H), 3.2 and 4.4 δ (3H), 2.75 δ (2H), 2.0 δ (4H) and 1.1δ (6H).

Example 75 N- [1-carboxy-2- (c) -amino-3-phenylpropyl] -O. Laipanyl-L-proline.

2-Amino-4-phenyl-3- (c) -3-phthaloylamino butanoic acid, obtained in Example 74, was condensed with pyruvoyl-1-proline via sodium cyanoborohydride as described in example 16. to give N- [1 carboxy-3-phenyl-2-phthaloylamino] -O _; L-alanyl-1-proline as a mixture of isomers. Heated to reflux this product with a hpdoazp ^ in ethanol for ¹ .5 h shlazhda was filtered off i-e. · .. '. g., I / - l seen to isolate the desired - .: ^t January product by flash chromatography and to obtain the title product of this example. After the following chromatography on silica gel and elution with ethyl acetate: 1 acetic acid. water is obtained with a spot of Rf C. 10 °.

Example 76. N-N-Carboxy-2- (3) -benzoylamino-3-phenylpropyl] D, L-alanyl-L-proline.

A solution of N-phthaloyl-3-amines-4-phenyl-2-hydroxy butyronitrile, obtained in Example 74, in ethanol, saturated with anhydrous ammonia, remained at room temperature for 3 days. The solvent is removed in vacuo and

BAD

ORIGINAL 1 is --ε c = your reflux residue for 6 hours is concentrated salt kN? The H-apssa was dried to dryness and the residue was purified with Dautex-5. ... bold, eluting successively with water - methanol

...- ipn _< 4, i. ··. &Lt; tb &lt;&gt;&gt;&gt; of 5 M ammonium chloroethanol The desired 2 3-diamino-4-phenylpropionic acid was isolated from the last eluate by concentration to dryness. A solution of the copper complex of this amino acid was prepared and the benzoylation of the 3-amino group in situ with benzoyl yl-pnd under alkaline conditions was performed according to the method described by R Aoeske et al. J. Am C.hem Soo., 78. 5883 (1956). The copper xyglux axis is broken down with sulfur medium and worked as described in the following literature to obtain 2-amino-3- (B) benzoylamino-4-phenylbutyric acid. The resulting intermediate was condensed with pyruvoyl-2-proline by sodium cyanoborohydride as described in Example 16 to give the desired N- [1-carboxy-2 (c) -benzoylamino-3-phenylpropyl] -D, L- alanyl-L-proline as a mixture of isomers of xylo can optionally be separated by chromatographic methods. Mass spectral analysis showed a maximum at 596 nrve for the disilylated molecular ion minus 15. The NMR spectrum showed absorption for two phenyl at 7.0 - 7.5 δ and diastesomeone methyl at 1.1 δ.

Example 77 N- [1-Ethoxycarbonyl-2- (5) -senzoylamino-3oenylpropyl] -D L-alanyl-L-proline.

When treating 2-amino-3-benzonylamino-4-phenylbutyric acid hydrochloride in

Vacuum afforded ethyl 2-aminopropo-3-amino-4-oenylbutyrate. Doo ko'- ^, day? 1 -oa ^and, on this intergl-nonpsdect with piovoig 6 -glghn by cyanosorhydrin isolation, as described in impurity 16, the title compound of the example can be obtained.

Example 78 N2-amino-1-carboxy-4-methylpentyl] - D. Lalanil-L-proline.

BAD ORIGINAL cm trans-3-amino-4- (2-metnlpropil) -2atss ^t: 'l,' ^O: derived moez adding hloosulfonilizotsianat to 4 ·. · .Gι ι. s <. j, .'- ab j> -r.ag / hom. who protected it as a tracer _; : methylphenyl derivative and then treated with lithium diisopropylamide, followed by tosylazide and chlorotrimethylsilane After acid treatment and chromatography with silica gel, grams-3-azido-N-N-methylpropyl; -e-azetidinone is obtained. which is hydrogenated (NO% d-q C ethanol) to an amino derivative and to 4.58 g benzylpyruvate in 20 ml absolute ethanol. contains South Molecular Sieves 4A. of sodium cyanoborohydride (0.65 g) in 8 ml of absolute ethanol was added dropwise. stirring until the reaction is 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 HCl to pH 3. The mixture was then adjusted to pH 9.5 with 10% sodium carbonate solution.

The aqueous solution was saturated with sodium chloride and saturated. extracted with the ethyl ^m empowered (5 x 40 ml). The combined organic layers were dried with Na 2 SO 4 and concentrated to give 4.94 g of an oily product. Chromatography on silica gel (ethyl acetate) afforded 1.11 g of purified product. The NMR and mass spectrogram are in accordance with the structure / composition of the benzyl ester of K ^! -trans-4- (2-methylpropyl) -2-oxo-3dZrumgg-1M ni-D gn.q QQ EXPLOITS ^υ ΟΘ3

..... _{h 4} , -r-. , »-S ς. ~ Q 'dtr fi G * n' f * ⁴ t; γ * Λ u raztess c 0 ° the acid (428 mg) and L-proline-t, Push ~ is: ^_ ec ί ?? 7 mA in 5 ml dime'il <Yormamid treated with a solution of e (605 mg) in 5 ml of dimethylformamide and after the test with 5 ml of dimethylformamide) in the hour, the ice bath was removed and the stirrer overnight at t / acetate (100 ml) was obtained and the resulting

Yu, t. - o / il. Triethylamine solution (223g boiling point for 20 min. After three mg of PP solution was washed with carbonate 13 x 30 ml) and water (1 x 50 ml of sodium bicarbonate solution was dried over sodium sulfate Concentration gave an oily liquid, 0 78 g,

whose I AM. ^p and mass spectrum are in accordance with the structure N-trans-h (2-methylpropyl) -2-ox 3-azetidinyl-D L-alanyl-L-proline tert <>

. The reaction mixture was concentrated to dryness at room temperature for 2 to 5 hours and the residue was treated with 1 N NaOH (30 ml) for 4.5 hours at room temperature. temperature. The alkaline mixture was slowly added to the highly acidic ion exchange resin and the product was extracted with 2% pyridine in water. Sublimation drying gave 0.30 g of N- [2-amino-1-carboxy-4-methylpentyl] - D. L-alanyl-L-proline. which consists of four diastereoisomers (S, S, S, S; S.S.FLS: R, R, R, S; R, R.S, S). which can be separated by chromatography. The MRI and mass spectrograms show structure compliance. The NMR spectrum showed multiplets centered at 4.5, 3.85, 2.3, 1.79 and 1.16 ppm. The mass spectrogram has a maximum at 458 (dissolved molecular ion minus 15 /

Impurity 79. N-12-amino-1-ethoxycarbonyl-4-methylpengyl] -Dalanyl-L-proline.

The intermediate of Example 78 N-trans-4- (2-methylpropyl-oxes-c-azetidinyl-D. 1-alanine (125 mg) can be condensed with L-proline, gasoline ester hydrochloride (167 mg) in the presence of dphenylphosphorylazide (191 ν '· and triethylamine (140 mg? in dimethylformamide to give 217 mg N- (trans-4- (2-methylpropyl) -2oxo-3-azetisulfonyl) - D. L-alanyl-E-proline) The benzene protecting group is removed by hydrogenolysis and the lac * lam can be opened with anhydrous sodium ethinate in ethanol to give N-12-amines-1-ethoxloroonyl -; mb

L-alanyl-L-proline

Example 80 N- [2-Benzamido-1-carboxy-4-methylpentyl] apanyl-L-proline.

N- [2-amino-1-carboxy-4-methylpentyl1-D, L-alanyl prepared according to Example 78, dissolved at the base of

p.

D.

D, LL- proline, treated with

- .f— - / * ps • ect. ⁿ : ^l lnra ^and no • X C

SNA

N - 2-benzamido-1 * ethoxenecarbonyl-4-methylpentyl] - D.

L-allanl-L-proline.

of a solution of N '- [2-a-lno-1-ethoxycarbonyl-4-carbon solvent with benzoyl chloride, can be obtained • dc-1-exoxycarbonyl-methyl-pentyl-O-E-alanyl-b proline.

Example 82. N-α- [1-Ethoxycarbonyl-3-phenylpropyl] - LL-proline.

Arginyl Work was done as described in Example 37 by condensing the Ν-α-tert-Soc-N-fo-HHTpo-L-apruHHH, the N-hydroxy succinimide ester, with Lprlip in dioxane water and the група-α-tert.-Boc protecting group. is removed is a acid. the dipeptide is isolated as is

Ethyl 2-oxo-4-phenylbutyrate linked to the resulting product as described in Example 25 is isolated as described to

POLUNES OF CODE C <Ό-α30 ^: In δρΓΗΗΗπδ, the BC is C ^ is PROTECTIVE with the pYR of the group.

This protection is removed by catalytic hydrogenation over &lt; 1 &gt; /. At κι C &lt; 3 &gt;; jars in vacuo to obtain a mixture of isomers of Ν-α- [1-3-phenpyl] -L-aop. '^and;'lPOOLNN | Gl-1Mer 83 Ν- (ί-ϊ1-ΚΑρ6θΚθΜ-3-φσ.Η1 ·! Γ.ρθ “; ·· Π]

L λ-Jr. '

N * (o -epsylO | \ »> n-caroonylp1 ^_ and,“ 3 ογιυκνΉcarbonyl-0 _: μ-homolysin. Concentrated by Homslide through the Copper Complex, with L-Proline 3 ° Butyl Ester can be then remove the 3 ° with three.Fluoric acid and the product can react groups can be η OWGlNAL ·· <*

5T _ _and -j ---. K0'- ^, D- ^, ZZAC1! C'HN ^, . ^, 1 I ^t item

: '· <R>_1; - _IG and n ₂ about by the ka-alntchmo hidoogenmrane. so to get · '

ПΛ

The intermediate described in Example 83. N-N-benzyloxycarbonyl-D-homolizin-L -proline can be reacted with ethyl 2-oxo-4-phenylbutyrate and NaBH 4 CN. The condensation product can then be hydrogenated over palladium on charcoal to give N, N -carbonyl-3-phenylpropyl, L-homolizyl-L-proline.

Example 85. N-α- [1-Carboxy-3-phenylpropyl] -β-amino-0. Lapanyl- L- ⁿ rollin

DL-α β-diaminopospionic acid was reacted under alkaline conditions with excess benzyloxycarbonylchloid to give blind acidification D _; La p-di (benzyloxycarbonylamino) -propionic acid, m.p. 123.5 - 124 ° C. Phosphorous pentachloride is added to. chloroform solution containing the di-Cbz product to obtain after treatment 0.1_-4- (bechyloxycarbonylaminomethyl) oxazolidin-2.5 dione Rzzozo of L-proline tert-butyl ester in methylene chloride is added to

tetrahydrofuran at -60 ° C. the mixture is distilled to the crude yielding the crude product. The trifluoroacetic acid effect breaks down tert-butyl ester in two hours at a glossy-g.: .--; for. ~ - is a liquid mixture of L prslin and N-benzyloxycarbonyl 'lamino-O-alanine-proline. By gel ^and ane 'column LW-20) pure e is obtained peptide. It condenses

2-Oxo-N-phenylbutyric acid and benzylcarbonylamino-D.L alanine-α-proline in the presence of sodium cyanoborohydride. Removal of the protecting group from the resulting product is obtained

p amino-D, L-alanyl-L-proline. NMR in

O ORIGINAL

.. J

D 0 gives aromatic subtones at 7.4 δ methic protons at 4.2 and Μν .-. Π'Π.πθ-Γ.! gg · 3 7 3 2 2 ^ϋ and 2 10 δ

.. '~ ~ ~ · r' - A ι_ι ι ι π r ·· r ~. G »η ι ι · 'I. P,. e. and m ui - O

........ - - · '· - y - ·. <· F- J f.

L- cuidr-H: - L- Proline

Solution of 3-Benzyloxycarbonylamino-D L-alanyl-L-proline. obtained as described in Example 85 and ethyl 2-oxy-4-phenylbutyrate ••; condensed into a solution of ethanol with NaS '- yC' The protecting group can be removed from the product by catalytic hydrogenation, so as to obtain Na-fl-ethoxycarbonyl-phenylpropyl] -5-amino-C. L-alanyl-L-proline.

Example 87 N1-ethoxycarbonyl-3-phenylpropyl] -O, L-alanyl-L-prel; ή.

Hydrolyzed ethyl 2-oxo-S-p-tsianofenilpropionat obtained in Example 72 by stirring in 5% sodium hydroxide at room TEM ⁿ era ^_ pops? ^d, ood ^h 'is ^and '. ^, is ^w a single knife. The reaction mixture was taken up. ; ether, the water layer was acidified to pH 2 with conc. hydrochloric acid, the product is extracted with a mixture of ether and ethyl acetate and the solvent removed to give 2-oxo-3-cyanophenylpropionic acid. Binds reducing this acid benzyl ester ^m and L-homsfenilalanina in the presence of d; fi: LR (Τ 'Li, 37 Ή (_ / ».» ^: ί *''* 4 · f \ Cl Λ. · U cl ·; - - · ~ *.' · ->, C. i'l L, ...

riOcMkiCTBanc a mixture of diastereoizsm ^ eras of N-: * - (£>. · etoksikarbs'nnl-d-fennltss ^_. _d! s L- p-tspanofenilala ^ in. The product is condensed with benzene, L-poolinat hidrohlorgd in dimethylformamide using the dipepylphosphorylazide reagent as described in Example 13. to obtain a mixture of diastereoisomers of N-. = - 5. benzene ester; - carbethoxy-3-phenylpropyl] -O. L-p-cyanophenyl -an / l -'... • '•• oenn. Hadsrsgennra this intermediate in ethanol is used to rum. м' m and ka i, lOd! op palladium on the outer and is formed as is described in Example 72 to the desired product is obtained in the form of a mixture of isomers.

| BAD ORIGINAL

Example 88 N- [x- [1-1'8 '- carboxy-3-phenylpropyl] -O. L-p-Aminomethylphenylapanyl-L-clone.

- SALT POLL \ / ct In Her

ΖΝ "and ^m pHe ° hydroxide in a mixture of. Ci. / t- · .in-t-kenpe overnight.

The solvent was removed from the reaction mixture in vacuo, hydrated, the residue in ethanol solution of hydrogen chloride and treated as described in Example 72 to give a mixture of diastereomers of the desired compound.

Example 89 N-α- [1-ethoxycarbonyl-3-phenylpropyl] -N-acetyl-1-lysyl-L-proline.

As described in Example 25, ethyl 2-oxo-4-phenylbutioate is coupled to N-acetyl-L-lysyl-l-proline in ethanol-water solution in the presence of sodium cyanoborohydride. Isolate on Dautex-50 as described and freeze-dried fractions of the product to afford a mixture of N- [1 ethoxycarbonyl-3-phenylpropyl-1-K isomers ^| -g> atce ^t ! D1-b lysyl- L- rot. Mass material. has a maximum at 653 (M648 η v-h,.,, ll. _{ _ '7 1-- «/! t M' L | ^ · j- | .1 ·. A D p,. 'r-: Λ r? · «'I_-, -pOj! IN.

Like t 3 is described in 26ίΜβρ 26. binds ethyl-4-oxo-4- phenylbutyrate ο L-histidyl- L-Proline in the presence of sodium  1; '? rn * L> Π vjn • I i! Λ, '' ¹ 'ii »· ι t / /'.. H ». U 'Λ H / C Λ L 'l o r C Λ instructions to obtain oooismeers of Ν-α-ί ι -etox ;; carbonyl-3-oenyl .-. ropig-b

* Istidyl-L · POLYLINE.

Example 91 A. N-α- [1-Ethoxycarbonyl-3-a-naphthylpropyl] -1-lysyl-L-proline

Ethyl 4-l-naphthyl-2-oxbutyrate obtained by alkylation of ethyl ι 3-dignai-2-carboxylate with 2-α-naphthylethyl bromide and subsequently

BAD ORIGINAL

And —-µ.

absorbs ·· ', a highly acidic ion exchange resin, and is eluted with 2% pyridine in it.

-UH A-rt t G1G- | G \ D \ G91 1 πΊ. I .. POMN. I _

d. ί \ - <. χ-ι ι-carboxy-3-a-naphthylpropyl-L-lisyl-L-proline.

Suspension of 4-a- ^and aftyl-2-scomaspe ^: -'2 acid ¹ - ?, ether by hydrolysis, adjusted to pH 7 with water with dilute sodium hydroxide, and dried by sublimation. The residue was treated with ε-3 ^ϋ - 6- ^{ethoxycarbonyl} -E-lysyl-1-proline. how is described in Example 91

A, to give N-[alpha] - [1-carboxy-3-a-naphthylpropyl] -1-lysyl] PP00, 1 L

Lproline.

{D \

·. V / chloro-e .-: yl-2-oxobutyric acid (1.1 g) in 5 ml of water was adjusted to pH / with a dilute sodium hydroxide solution and treated with 0 07 g of NaSH ₃ CN in 1 ml of water for several hours ^and aooa . After stirring, place the room / Ί ·· in water to give 0 0 8 g of product. NMR indicates that the tert-oct protecting group is not j-ι η n m - 0 0 0

- .Ki G-i ο <- - · 0 - - - <o -> -; •• 'G I 09.C · L ίJ S. ** m_

m. <

1.

IL- L- PSOLES ► * ΓΛ

I'm not with the structure. A molecular ion was found at 584. by chromatography

PSL.in hso <c l \ 81 ίαπι ί Iss

..; _<t ; ORIGINAL. - - I * ·

Ί ^ ΗΗΜί.Ι · '.') .N.ynf> nA ^ unn] .i ko ^, .'kaosn :; ^, l -_- l; ¹ Zil ^! _- ~ ^roll,.! n in ethanol solution with excess izil- l- poslin.

Solution

Ν-α- [1 -carbox. 1-3- (3,4-dichlorosulfonyl-lropyl] -L lysyl-L-preline.

of 4- (3 _: 4-dichlorophenyl) -2-oxobutyric acid obtained from dichlorodihydrocinamate by condensation with ethyloxalate and subsequent acid-catalyzed hydrolysis and partial decarboxylation in water was treated as described in Example 92 to give M-a - [1-carboxy-3- (3,4-dichlorophenyl) -propyl] -1- lysylL-proline. The NMR spectrum at 0 ₂ 0 shows aromatic hydrogen at 7.30 ό, wide singlet at 4.3 δ and broad multiplets at 3.53, 3.00, 2.68, 1.98 and

6o L. The mass spectroscope shows a maximum of 775 m / e. which corresponds to the loss of a vest from the desired structure, a large M - 2 (457 gp / e) isotope contribution confirms the presence of 2 C !.

N-α- [1 (5) -carboxy-3- (3-indolyl) propyl] -N lysyl-L-proline.

- 0 _ p f · -

V 'ho.otriptophan by the method described by Weygand ef al ..

App 658.

ι it (1962) Condensed with-tert-Boc -? - lysyl-1_-proline in sodium cyanoborohydride, as described in Example b4, for the presence of a crude mixture of diastereoisomer of | -carboxy-3- (3.diolsGprspkjarGntret.-Voo-E-lzll-d-roll. g. The spectrum of the tetrasilylated product has a maximum at 732 (Mg). 717 (Mg-15). 615. 660 and 645 m / e ^P azrushava is the protection of the lysine side chain chsez treatment with 4 N-hydrogen chloride in the ethyl ^m empowered and purified with a strongly acid ion exchange resin as described in this example, in order to, _n · "and

/ * \ ν Γ. Ξ Η from · '^; : 'od' / kt noe staining is obtained

.'omeo.

 '' '- ο • - • cp ^: _KO .-! LslS »: Lg_- LYSYL-! _ GROLIN.

Benzyl 2-oxo-7-u ^m alimidoheptanoat and ε-3 ^υ - butoxycarbonyl-L ·.-Proline are ks.-condensing with an excess of NaBH ₃ CN b ethanolic oaz'5oo to obtain Na-fl-benzyloxycarbonyl-bSled this can remove the onsyl group by hydrogenation over palladium, remove the tert.-Boc group with 45 N HCl in ethyl acetate, and remove the phthalimidogroup by hydrazine treatment to give

N- [x- [6-amino-1-carboxyhexyl] -1- lysyl-L-proline.

Example 97. N-α- [6-amino-1-benzyloxycarbonylhexyl] -1_-lysylL-proline.

In the treatment of N- [alpha] - (1-benzyloxycarbonyl-6-phthalimidohexyl) -N-8-3 ^with '-butoxycarbonyl-l-lysyl-l-proline. prepared according to Example 96 with 4.0 N HCl! in ethyl acetate and then with one equivalent of hydrazine in ethanol, under reflux, a mixture is obtained from which N-N-benzyl-sicarbonylhexyl '* -j-lysyl-L-poslin can be isolated.

lysylprslin.

Benzyl 2-oxo-6-phthalimidohexanoate was treated as described in Example 96. to give N- [5-amino-1-carboxypentyl] L-l-3 / l-l-carbon black chromatograph ^The desired product is at 516 m-e, M'-15 ion at 510 m / e and a maximum of 399 m / e. which is indicative of the loss of silylated carboxyl.

Example 99 o-amipd-1-oennlox. more sorry · - ·. ppm = / ί-L- proline.

J 'AND

R αι_ιθ | 1D

2-oxo-6-0 thalimidohexanoate

as described in Example 96. except that desc-.ί is eliminated! '.'Have hydrogen on palladium From the product mix

1 1 Q * ~ 'dd Μ-, γ-Γ 1 .Κ ^ οόοκοίЧС <' ^ ^т ' ^{l n} ' ~ I hc. , _ Proline.

Phenoxypyridic acid <0.9 g) is dissolved in water, the pH is adjusted to 7 with a dilute sodium hydroxide solution and the solution is subjected to sublimation drying. The residue was dissolved in 10 ml of ethanol and treated with N - 8-3 ° -butoxycarbonyl-1-lysyl-1-proline (0.36 g) and powdered molecular sieves No. 4A (3 0 g). Sodium cyanoborohydride (0.13 g in 3.5 ml ethanol) was added in portions 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 eluted with 2% pyridine in water, followed by freeze-drying to afford 0.25 g of product free of protecting groups групи-α- [1carboxy-2-phenoxyethyl] -2- lysyl-L-proline. NMR and mass spectrum are consistent with the structure

5. N- [alpha] - [1-ethoxycarbonyl-2-phenoxyethyl] -L lysyl-L-proline.

E ^m ^ yl phenyl oksipiruvat treated with E-3 ° -butoxycarbonyl-1_saglaspo Example 100 A to afford the Ν-α- [1lro PF> n spectrum shows absorption phenyl.

centered at 7.3 δ, and wide proline.

was treated with c-3 ^is -butoxy carbonyl-E-lysyl-L -proline as described in Example 100 to give N-α- [1-carboxy-2-phenylthioethyl] -1_-lysyl-L-proline. The mass spectrum shows a silylated molecular ion at 567 m / e. Atheroscopy yields the desired isomer.

ii .and I i • 'u, r <μι ·,,,,. ,. I; -; and 7 f | i _ r. Γ, .: 1 Π _; ¹ W> · <- / k Γ \ Va sz 4z I '»'? I γ vl. ''> · .4 / v. I 7 ': L_, Λ. k> I L.

r * ·; > · _ ”·‘ · ’’ - ·. '=: -3441, _se ΤΟ βν _; , ..- _ν - ρ-. ^ £ .and _S gg- icyeer .-: · -3-sutoksi • 'S.obonpl L-ri'Ciin-L-nponnH under ^d Simer 100 ^(A; to give N- [1-ethoxycarbonyl-2-phenylthioethyl] -E-lysyl-L-proline.

Example 102. N-a- [1-carboxy-2'c) -phenylpropyl] -01- lysyl- Lπροπ ', Ή.

N-N-amipO-4-phenyl-O- (hortalim / _l dooutanoate) is condensed with 2oox-6-phthalimidohexanoic acid (obtained by alkylation of benzyl 1,3-dithian-2-carboxylate with phthalimidobutyl bromide and followed by oxalidobutyl bromide) hydrolysis) in the presence of sodium cyanoborohydride as described in Example 13. The resulting intermediate was coupled to the L-proline benzyl ester hydrochloride by diphenylphosphorylazide as described in Example 13 to obtain a mixture of isomers of benzyl ester. of N- [alpha] - [1-ethoxycarbonyl-2 (S) phthalimido-3-phenylpropyl] -1H-o-phthalenyl-O, [_- After treatment with two equivalents of sodium hydroxide in a solution of ethanol-water for 4 hours at room temperature, neutralized to pH 4 with concentrated hydrochloric acid, the ethanol was distilled off in vacuo. The product is C Ks, I ka Sa. · ' ¹ '»<a ca S '>' U C U C i G I p Sk ύ ί Ο> _ sa, C. ..-., Sa ZZ L ·. V The 3 ^ -0 3 <lK y, 'w!. J; ed is heated with ocoaiCn x. ^ Admixture in ethanol containing 2 equivalents of hydrazine. for 1.5 hours and isolated as described in

! measure 1 and 3.

Ν - (/ - ί and -carboxy-2'-oenzoylamΗθ-ό-ψеНи1лΓΊρ / ΛΓίΜημΡ i.! - ^Γ Ό0'ΊΗΗ

Example 7 is condensed with 2-oxo-phthalimidO-hexane acid in the presence of sodium cyanoborohydride by the method described in Example 13

The resulting N- [alpha] - [1-carbethoxy-2 (S) -benzoylamino-3-phenylpropyl] - N - with phthaloyl-0,1_-lysyl can then be coupled to the hydrochloride of g

BAD ORIGINAL urine-1'-Phosphorylazide as

GC. * 4 lysyl-1_-proline purification by chromatography Poi with the equivalent of sodium hydroxide in ethanol solution for hours at room temperature

102 gives N- [alpha] -N-carboxy-2 (S) -benzoylamino-3 phenylpyridin-yl] -N-phthalyl-O ₁ b lysyl-L-proline. By heating the latter to reflux in ethanol for 1.5 hours in the presence of one equivalent of hydrazine and isolating according to Example 74, the desired compound under the farm of a mixture of isomers can be obtained.

Example 104 1- {1- (x- [2-amino-1-carboxyl-1-4-methylpentyl] -OH b lysylL-proline.

The trans-3-amino-4- (2-methylpropyl) -2-azetidinone ethanol solution prepared according to Example 78 can be reduced reductively with benzyl 2-oxo-6-phthalimidohexanoate using sodium cyanoborohydride and molecular sieves. The product Ν-α- [4 (2-methylpropyl) -2-oxo-3-azetidinyl] -N-phthaloyl-0,1_-lysine benzyl ester can then be debenzylated by hydrogenation over palladium.

The free acid and the gasoline ester of poolin bind to the lp τ, '. Οο n-pr''ΖΠ- ^t ~ ри ⁿ obri9np; <pa

WJ 1 VU CHK _ ^: -4. ¹ UV / C Jf 'I <4 i X- / H «x _m U P- ~ I I);

\ Ca1 \ IV described above to obtain N-α-U2-methylpropyl ^: 1) -2-oxo-3-

The otaloyl group is

N-a-4 <2-methylpropyl] -2-oxo-3-a-thiol base

G.- ⁴

BAD ORIGINAL ⁿ wp 105 α · α · [2benzamido, ./zyl- L- proline.

- k a p b ο / ’'- 4 - m e t i l p e h t i l] - D. L -

2-oxs-2-azetidinyl-3-C3-oxo-carbanyl-DL-benzyl can be obtained from trans 3-amino-2- (methylpopyl-2-azetidione (impurity 78) and benzyl e-3 ^L '- butoxycarbonylamino-2-oxohexanoate. The gasoline group is removed by hydrogeneration and the product is coupled to the L-proline gasoline ester. methylpropyl] -2-oxo-3azetidinyl-N-c-3 ^O butoxycarbonyl-DL-lysine is then debenzylated with hydrogen and β-lactam is hydrolyzed with a dilute base to give N-a-4- [2-amino -1-carboxy-4-methylpentyl] - Ν-ε3 ° 'butoxycarbonyl-DL-lysyl-E-prol After benzoylation with benzoyl chloride in an organic solvent, the protective tert.-Boc group is removed with trifluoroacetic acid to give N-α- [2-benzamido-1-carboxy-4-methylpentyl] -0, E-lysyl- L-Proline.

Example 106. N-α- [1 (3) -carboxy-3-β-chlorophenylpropyl] -Lisyltrans-4-methoxy-E-proline.

The hydrochloride of non-methyl-trans-4-methoxy-prolineate is coupled with Ν-α-tert-Bcc-N-c-Cbz-L-lysine using dicyclohexylcarbodiimide and triethylamine in methyl ethyl chloride.

as described in the primer is purified by chromatography. the ester is hydrolyzed and the protecting tert-Boc group is removed as described in the same example. Reduction of N, N-8-C6-E-lysyl-E-4-methoxyproline is reduced

2-oxo-4-o-chlorophenylbutyric acid (obtained from p-chloroxyl ethyl ester: alkaline-catalyzed condensation with d,: ethoxalate, followed by decarboxylation in anhydrous hydrochloric acid in acetic acid) presence of sodium cyanobo * idride and subsequent treatment according to Example 24. to obtain a mixture of isomers of Ν-α-11-carboxy-3-β-chlorophenylpropyl] - Ν-εCbz-L-lysyl-trans-4-methoxy- L-Proline BenzyloxyumiNAL is removed

..__

O / kars / trapping by catalytic hydrogenation and palladium-on-charcoal catalyst in the usual manner. By means of raffi =, the desired mass of the ectog is maximized at 584 »M '- 1). 5s8 - 15). 466.397. Oats and 352 (base maximum) for the disilyl product

N * and - [- carboxy-3-p-chlorophenylpropyl) -L lysyl-4-p-azotidine carboxylic acid.

estes

N, N-Cbz-L-ruCHH with 1-thiazolidin-4-carboxylic acid hydrochloride, the product is purified by chromatography, the ester is hydrolyzed and tert.-Boc protecting group is removed by the methods described in Example 20. By reducing coupling with 2-oxo-4-chlorophenyl butyric acid as described in the same example, a mixture of isomers of Ν-α-ΓΙ-carboxy-3-p-chlorophenylpropyl) Ne-Cbz- L-lysyl - L-thiazolidine-carboxylic acid. By removing the benzyloxycaobonyl protecting group when treated with hydrobromic acid in acetic acid at room temperature in the standard peptide chemistry manner, distilling off the solvent acre-freeze-drying.

S'oS

--1 ,.

mo 108 \ '- R7- [1-carboxy-3-p-chlorophenylpropyl) -L' lysyl '*' ethyl esto * and p-ans-5-methyl-thiazolidine-4 thioacetamide. acid reaction with fsmaldehyde, hydrolysis to β-methylcysteine and following the method used by R.F. Nutt et al.

Abstracts of the 6th Am tert-Bcc-N-c-Cbz-L-OHSHH were combined using 2.8 ml of triethylamine and 4.5 g of dicyclohexylcarbodiimide in methylene chloride as described in Example 20 by reducing the coupling of this intermediate

Vr · c 2-oxo-4-p- [llorophenylbutyric acid using cyanoborohydride and protecting the benzyloxycarbonyl p- · ₁ ροπι · ιπ. · <- lysyl- and _- trans-5-methylthiazolidine 1 cassic acid in the form of ^Li a mixture of isomers.

Example 109. N-α- [1-carbxy-3-p-chlorophenylpropyl) -1_-lysylL-3,4-dehydroproline

The L-3,4-dehydroproline ethyl ester hydrochloride is condensed with Ν-α-tert.-Boc-N-8-Cbz-L-nn3HH, the third, Boc group is removed with 4 M HCl in ethyl acetate, then bound reductively the intermediate with 2-oxo-4-p-chlorophenylbutyric acid, the protecting group with hydrobromic acid in acetic acid was removed and further treated according to example 107 to give a mixture of isomers of M-α- [1-carboxy -3-p-chlorophenylpropyl) -1_-lysyl-L-3.4 dehydroproline.

Example 110. N- [1-Carboxy-4-methylpentylN-alanyl-L-proline.

A solution of 5-methyl-1-oxohexanoic acid (1.44 g) and L alanyl-L-proline (0.37 g) in 5 ml of water was adjusted to pH 7 and the temperature of the reaction product was treated for one day.

S '

O O / η 1 L AM Π I L LJ

P-ν 'b> jJ ν »m' · to obtain 0.5 g of product after freeze-drying

One part (0.2 g) was purified by column chromatography on LH 20 to afford 18 g of 1-carboxy-4-methylpentyl-1-alanyl-L-proline. NMR μ? Ββ ~? Κτ.; ~ Μ · are in with ^m ? E ^m with ^m ? '' ¹ is with the niastereoisomers

'. * o ^g a' · be isolated by chromatography. Peaks of 443 (M ~ -15), 458 (ΜΊ. 341.301 and 244 m / e (baseline maximum) are observed in the mass spectrum of the product being distilled.

^g Example 111 N- [1 (R) ethoxycarbonyl-4-methylpentyl) -ts- alanyl-L-proline.

BAD ORIGINAL: -: - 1.7> Mesh-2-oxo ⁱⁿ exanoate (3 44 g 'n L-alanyl-L-proline (0.74: s) is mixed with 15 ml ethanol with 6 g powdered molecular sieves 4A. E. - Pa-iKuj ik-j hours is added dropwise to sodium cyanide. dr1-! d id io ~ <=. eh-s. ^- h, - then the anol is removed in vacuo the product is absorbed on a highly acidic ion exchange resin · eluted with 2% pyridine in water to give 1 08 g of N- [1ethoxycarbonyl-4-methylpentyl) -E-alanyl-L-proline One part was purified by LH-20 chromatography for spectral analysis. The NMR is consistent with the structure. The mass spectrum shows a maximum of n and 114 (silylated molecular ion-15). By chromatography to give the desired izsmer.

EXAMPLE 112. N -41-Carsxy-3-p-phenoxypropyl) -1_-alanyl-L-proline.

Mixture of 2-oxo-4-p-phenoxyphenylbutyric acid, obtained by the reaction of p-phenoxyphenyl Grignard reagent with ethylene oxide daza alcohol, which is converted to bromide and condensed with ethyl 1,3dithian-2-carboxylate by oxidative cleavage .

followed by alkaline hydrolysis. The resulting ketoacid and Lallall-L-proline with zoda at pH 7 adjusted with a dilute base were treated with excess sodium cyanoborohydride. The product N- (1karboksi-3-p-Phenoxypropyl '- (_- alanyl L-proline ^m Seeli ex': sleev ^item of dnsililnraniya (M) obZ (M -loj, 468 i.o / 0 rii / e

N '- (1-Ethoxycarbonyl ο is isozoa at maxima at

Psymer

584

113.

product contains (baseline maximum).

-3-p-phenoxyphenylpropyl) 2-oxo-4-c-phenoxyphenylbutyric acid, as

Oh! 1-!

ntslmer 112. with the exception that h may oyde omitted lralna alkalpahidosliza and condense L- alanyl-L-proline in grisas ^d velopment not a sodium tsianoboohidrid so as to obtain N- (1o-rksiksrbsnpl -3-p -phenyxiphenylpropyl) - L-alanyl-L-proline.

Example 114 M- (1-Carboxy-3-phenylpropyl) - alanyl-D, ί

3,3-dimethyl-proline.

BAD ORIGINAL

3-Dimethyl-P-L-prslin is obtained from 3-methyl-2-butenal by the Cox method _: J. Chem. Soo .. 1964 5024. and converted to the hydrochloride of Mg · .K, l;. ·? c'-Tup - _ · /. _. ·. ~ s; _ Д ДВДЗсс; This is condensed with n-oxo-4-phenylbuty * one as described in Example 47 to give a mixture of the isomers of the desired product.

Example 115. A. N- [1-Carboxy-3-phenylpropyl) - N - (3Csylcysteinyl) - N -

Condensation of L-N-tert-Boc-benzylcysteine with L-proline tert-butyl ester in the presence of dicyclohexyl carbodiimide in the usual way gives a blocked dipeptide L- (N-Tper-Boc-Sbenzylcysteinyl) - L-proline-butyl ester. The latter was treated with 4 N HCl in ethyl acetate at 0 °. to give P (3-benzylcysteinyl) -k-proline. Treatment of this dipeptide with 2-oxo-4-phenylbutyric acid in the presence of sodium cyanoborohydride yields N- (1-carboxy-3-renylpropyl} -0-1b-oenzylcysteine * proline as a mixture of isomers.

B. · N- (1-Carboxy-3-phenylpropyl) -k-cysteinyl L-proline.

N-carboxy-3-phenylpropyl; -N- (5-benzylcysteinyl) -B-proline obtained in part A. is tautoated with sodium in liquid ammonia to give the desired compound.

Example 116. N- [alpha] - [1-carboxy-3-phenylpropyl] -k-ornyl-L-proline.

N-tert.-Boc-k-ornityl-proline and 2-oxo-4-phenylbutyric acid were condensed in the presence of sodium cyanoohydride on rci ~. The protecting group is removed from the product in ethyl acetate. The crude was adsorbed on a strongly aqueous solution of 3% using 4 × hydrogen chloride gas. The 2-way hydrochloride salt was acidified with a resin and extracted with pyridine. The mass spectrum shows a molecular ion at 355 m / e for the product - 36. The NMR spectrum is consistent with the structure.

BAD ORIGINAL

L_.

Χ-ε- ^τ Ω? Τ - BOSS-L-lysyl -'_ · • Ι .- 'ΖΛ, 7. Γ 1; -Y. Γϊ-Γ »u / - 't. ·” · Γ · _.4ι Qu! L Π Γ>Γ> ζλ π ι 1 π ·· - '''·''*I> - · -' · - '· - ~ f- - - - - Τ -''*-.' · Lazyl-L-proline

Z- .-. Z-h :. -and, ·. b-t  'a * '} -

to which are added ^1. '

a .- \ ν on is towers! c .. ..>,> -.after sieving the reaction, sieves are removed by filtration. After evaporation, the residue from the filtrate is dissolved in water, extracted with ether and absorbed on a highly acidic ion exchange resin. Elution with 2% pyridine in water gives 639 mg of the crude protected product M-a- [1 (B) -ethoxycarbonyl-3-

The protecting group was removed with 4 N hydrogen chloride gas in ethyl acetate. The resulting HCl salt was absorbed on a highly acidic ion exchange resin and eluted with 2% pyridine to give 270 mg of product. The mass spectrum shows a molecular ion at 678 lp / e for the distillated products plus 1. The NMR is consistent with the structure. Chromatography afforded the desired monomer.

Example 1'3. N- [alpha] - [1-carboxy-3-phenylpropyl] -N-c- [beta]-dimethyl-Elizyl-L-proline.

ester is alladic on charcoal, 27c * 1 CT · • fcl. · Hi - Γ * ⁴ * II. JO 'J 1 /: /. _t . .

> - ci p £ D j h ✓: U; and .λ

κΗτ.ι 'hydrogen pressure). ^{M-O-toet.-Bss-au:!} This group is cleaved with 4 N hydrochloric gas in ethyl acetate. As described in Example 54, 2Oxo-4-phenylbutyric acid and Ν-ε-N-ε-dimethyl-Lysyl-L-proline hydrochloride were condensed in the presence of sodium cyanoborohydride.

2, the pectrum is consonant

proline.

N-m-a- [1-carboxy-3-phenylpropyl] -1_-lysyl-L-proline, a mixture of diastereomers obtained according to Example 57 B, was purified by

BAD ORIGINAL

Ο _; -ι ^r * '·· “- <' ·· r · 42 D .'JCiTQuO n HAP = 2 columns prepared. a <t ^ is c-mica but ^{p is} named is calibrated at 53 ° C with 0 1 Μ H ₄ OH. * - -. _l , - _ - -.? k · - mo ^: '1' · and ^ talking in; With ml of the same time: gauge and added to the column. When

the elbows are eluted with this solvent, the first isomer appears in ci ·? · .'- · g shoes 320 - 360 ml eluate The second isomer appears in the range -icQ 540 m-l of eluate Me. ldn fractions do not contain a mixture of isomers . when the fractions contained the / first isomer. freeze-dried to give 72 mg of loose white mass. This is the rounder isomer ^m with the SSS configuration by analogy with the more active isomer of N- [alpha] - [1-carboxy-3-phenylpropyl] -1_-alanyl-L-proline. , which was found by X-ray analysis to have an SSS configuration. Thin layer chromatography on silica gel in 1: 1: 1: 1 ethyl acetate / n-butanol / water / acetic acid gave a single spot with an Rf value of 0.43. The 300 MHz NMR spectrum showed a triplet for the methine prot γ to the phenyl substituent 3.40 ppm. For sublimation SuL * jcnNci | DdK '' r with rci □. · Λ ». ^ Υϊ» ι c> ι <y όc * c * · ¢ - · * ι · solid. Thin layer chromatography of this substance gave a spot with an Rf value of 0.39 300 Mhz The NMR spectrum obtained showed a single triplet for

61 cc.

the methine proton γ to the phenyl substituent at ι by lysylbenzene.

as described in the example

54. as 2-oxo-4 acid and

N-acetyl-L-lysyl-L rat in the presence of sodium cyanoborohydride. ha

The NMR of carbsx-3-phenylpropyl] -. The 1-c-acetyl-L-lysyl spectrum shows a molecular ion at 663 for trisilylated products.

Example 121. N- [alpha] - [1-carboxy-3-phenylpropyl-N-arginyl-L-proline.

BAD ORIGINAL

ml is obtained by: DCC condensation of the hydrochloride salt of the L-proline gasoline ester and of the N-ot-tert ^P o ^g -. '\ - <o-us 'ro- L-arginch Ν-α-tert -Eos-protecting group is -, .- _o Qb ^{! 1} uRJ4 '! ~ Uoi.hiu -> .j NJHCl 5 ^Q T' ** 7 ° ^{! 1 with} r3 ^m n POLYL N ⁰¹ - ¹ ',' n-N-nitro-k-arginyl-β-proline benzyl ester was condensed with 2oxn-4-phenylbutyric acid as described in Example 54

The reaction yields a very low yield (25-33%) of M-α- [1-carbxy-

3-phenylpropyl] - N-ω-nitro-L-arginyl-L-proline. This compound (159 mg) was dissolved in a solution (2.5 ml) of acetic acid / water / methanol (84%, 8%. 8%) and hydrogenated at 276 kN / m ^{2 at} room temperature over 130 mg of palladium on charcoal. simultaneous removal of the ω-nitro protecting groups and of the benzyl ester. The catalyst was filtered off and the filtrate was evaporated to 94 mg, and the water-soluble portion was dried by sublimation to a loose white mass (90 mg). This solid is acetate of the desired product and is converted to the free base by absorption on a highly acidic ion exchange resin, washed with water, eluted with 2% pyridine in water by freeze-drying the product rich fractions to give 60 mg of M-a - [ 1-carboxy-3-phenylpropyl] -k-arginyl-L-proline. n '», - slzk ~ bM7 is in accordance with the structure. The mass spectrum shows the molecular ion poi 793 for the pentasilylrap products

Example 122. N- [1-Carboxy-3-phenylpropyl] -1_-histidyl Lproline.

It was worked as described in Example 54. The 2-eye-4-phenylbutyric acid and the n-histidyl-c-proline were condensed in the presence of sodium cyanoborohydride. to give N- [1-carboxy3-phenylpropyl] -histidine-kspin. The product was purified by gel filtration in methanol (LH-20). The NMR spectrum is consistent with the Coc structure. The mass spectrum shows a molecular ion at 657 for the disilylated products.

BAD UrtiGiNAL

u ....----—

I.

CX ^- G ι u <Sl ”7 D ι v * r. Ach 4 1 </ ΐ'ίΠ ^ ύ ,. 'ν'ί-κύ ^ r · or. ? r- c ·. b - 'ci - ι uj Ο κ'Ό and ^L -> Π Oui ι'»'l ~ - 4 .. ... χ, ^ ... · .4 h / 4 ^Γ 'ν>. - _Μ νΠ 4 »^ ν4 ~

Γ \ ι _ <“. Τηοτ _Ρ <χκ ^ _ΐ _ PI7IL. «Μ <> I Μ V ¹ . ϋ 4? 'Ζ 4_? I G »4-, ι · I

L-proline in the presence of sodium cyanoborohydride. The µ-tert-Boc protecting group is removed from the product with 4Ν HCI in epiacetate. The resulting hydrochloride

on Datex 50 (NM and eluted with 2% pyridine in water. Sublimation drying of product-rich fractions gave the free base salt of the light brown solids. The NMR spectrum was consistent with the structure. The mass spectrum showed a molecular ion at 718 for tetrasilylated

Example 124. N-α- [1-Ethoxycarbonyl-4-methylpentyl] -1_-lysyl Lproline.

Dissolve 2-oxo-4-methyl-ethylpentoate (2 75 g) and N-ε-τρβτ.-Βοο-Ι-lysyl-L-proline (2.75 g; in 150 ml of ethanol containing 16 g of powdered molecular sieves 4A. Hydrogenate at 276 kN / m ² room temperature, above 'g' 0% palladium on charcoal After absorption of 1 mole of hydrogen, it is filtered through a filter mass and washed carefully with the eia ~ cn catapitator on the filtered precipitate.

G τρνΜ ^η '' ^τ

Jn 4 K / JI 4J *. · ¹ 44 14- Π 4 ¹ Spend

mn) to remove neutral products. The pH of the aqueous layer was adjusted to ^7, saturated with sodium chloride and extracted with - - - - -,. ~ <0 in -1 G.H D R ST O <4 nL 'J? n r ·. p m-ITR - *> · '. ·· «-JS- ¹ , - - - - .. ^ cs.a. L.4.4,! <4 ^. '_; .4 · · _{ . · K! C. 4. NN anhydrous magnesium sulfate. The ethyl acetate was evaporated to give (138 g) of crude N-cis- [1-ethcC111 <arbonyl-4-methylpentyl] -1'-1-c-tert-Eos-11-yl] L-proline. Remove the protecting group tert.-Vos on the usual pa -,: .- with 4 NS! in ethyl acetate. The resulting hydrochloride was converted to the free base with a highly acidic ion exchange resin, eluting with 2% pyridine in water. Product rich Fractions are subjected to

BAD ORIGINAL k

ν y ο »«> -0.k «ui ^ r1ps drying. to being put forward hygroscopic crumbly> z_eo ^t d? NMR spectra I GL | .11 4bG1 1 L ^ ar <nr.v. '' _L , | ..t. »* UrunriGuTUr :. . .Πΐ'ΟιΛΓΊ I - ΓυΛΤΠΜφ

Ν- <χ- | 1 -etokoikaroopkign-1istilpep71) ptsi'Lizil L- proline are and i.oo HyiSi ii> a to the corresponding acid by karoonova .zhvane in CLPP pootrlr uia sodium ^{hirooko! 4R} 5 Etch? N ^o sn ° ntya ^(d, oi buil The reaction mixture was acidified to pH 5, absorbed on a highly acidic ion exchange resin and eluted with 2% pyridine in water. The product-rich fractions were dried by freeze-drying to give N-α- [1-carboxy -4-methylpentyl] -1'-lysyl L-proline as a white loose solid The NMR spectrum shows a molecular ion at 516 for the disilylated products.

Example 126. N- [alpha] - [1-carboxy-3-phenylpropyl] - [beta] -L-leucyl-LtrI and иαπ.

Work as described in Example 54. as 2-oxo-4-phenylbutyric acid and? -Leucyl-? -Triltophan are condensed in the presence of sodium cna-hydrochloride. The product is subjected to a sulfoxation mixture of dioxane? water, as it is poorly water-soluble NMR spectrum is compatible with the structure

M Q n O and y P O <-] i '

M0H Pr * /

695 for trisilylated

Tablets.

and drink it <nop / o nvr> vO λ i. 'ί m p * <c p Kry | _ | m Π - ^l LPPIL

1-allanl-1-prol .: -: (25 mg), pre-clustered starch (USj patent (82 mg), microcrystalline cellulose (82 mg) and magnesium mode Ν-, 1 (S'i-ethoxycarbonyl-Zmg ) can be formulated 3 - f e n and l p p with p l / - L - al l a n and l - L - π ρ o l i n with the composition of pre-clustered starch ic.

ί 'x ί L>

feniprs ^g Ф '′ − b − li311L-1_-proline (20 k I · 4 f O X * “-' f Λ <· </ GP” ~ H ΓΛ | '- u'., ό. r.ci on

BAD ORIGINAL microcrystalline cellulose and magnesium stearate as described above.

B. Combination tablets.

The combination tablet with a diuretic, for example hydrochlorothiazide, typically contains N- [1 (c) -ethoxycarbonyl-3-phenylpropyl] -1-alanyl-L-proline (7.5 mg), hydrochlorothiazide (50 mg), pre-clustered starch (82 mg) and magnesium stearate (1 mg). Tablets with, for example, N- [1- (S) -ethoxycarbonyl-3-phenylpropyl] -1-lysyl-1-proline (5 mg) and hydrochlorothiazide (50 mg) are prepared by first replacing N / S (c) -ethoxycarbonyl -3-phenylpropyl / in the composition described above.

B. Injections.

The injectable composition contains N71 (3) -ethoxycarbonyl-3-phenylpropyl / -1-alanyl-B-proline (5.42 mg), dibasic anhydrous sodium phosphate (11.4 mg), benzyl alcohol (0.01 ml) and water for injections (1.0 ml). Similarly, this composition can be prepared using N- [1- (c) -ethoxycarbonyl-3-phenylpropyl] -1-lysyl-1-proline in place of N71 (S) -ethoxycarbonyl-3-phenylpropyl] -Lalananyl-proline . The injection composition may also contain a pharmaceutically effective amount of another compound having an antihypertensive and / or diuretic action, for example, hydrochlorothiazide, as described in point B of this example.

G. Suppositories

A typical rectal delivery suppository composition may contain N71 (S) -ethoxycarbonyl-3-phenylpropyl / -1_-alanyl-B-proline (1 25 mg), butylated hydroxyanisole (0.08 - 1.0 mg), disodium calcium edetate (ethylenediaminetetinotetinaminotetinotetinaminotetinotetinaminotetinotetinaminotetinotetinaminotetinaminotetinaminotetinaminotetinaminotetinaminotetinaminotetinaminotetinaminotetinaminotetinaminotetaminotetaminotetaminotetaminotetinaminotetinamide acid) (0.25 - 0.50 mg) and polyethylene glycol (775 - 1600 mg). Another suppository composition may be prepared by substituting, for example, N- [1 (c) -ethoxycarbonyl-3-phenylpropyl] -6-alanyl-6-proline with N- (1 (c) -ethoxycarbonyl-3-phenylpropyl) -1_-lysyl-1-proline and by substituting, for example, disodium calcium edetate with butylated hydroxytoluene (0.04 - 0.08 mg) and polyethylene glycol with hydrogenated vegetable oil (675 - 1400 mg) as

BAD ORIGINAL ϋιϋΐυΐ). .it ί ν; ecobee PS. Wtcobee ivl Wttepsols, etc. Composition for contraception: 'thorns may contain other antihypertensive compounds

Kami with 01 ORIGINAL DOTS and with HP ι. TIMER

11 Example 128 N- [1-egoxycarbonyl; 1-3 (3 indolylpropyl)] - L alanyl-L-psoline

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 was hydrogenated with 10% palladium on carbon in the presence of 3.2 g of powdered molecular sieves (4A ) at a hydrogen pressure of 276 kN / m ''. After completion of the reaction, it was filtered and the filtrate was concentrated to dryness.

The axis was purified with tutC.U.7ixJ U / C ν ν / | \ and methanol and chromatographed on a Sephadex LH-20 column to give 0.67 g of a crude product in the form of an oily orange liquid. . A portion of the crude product is purified by dissolving in water and extracting with ether. The product is recovered by diluting the aqueous phase with an equal volume of saturated sodium chloride solution and extracting with ethyl acetate to give 80 mg of N- [1-ethoxycarbonyl-3- (3-indolylpropyl)] -1-alanyl-L-proline below the farm of yellow oil. The mass spectrum shows a maximum at 553 for the disilylated product and at 631 for the trisilylated product.

Benzyl N-1 1 (8carboxy-3-renylpropyl] -1 alanyl-L-prolinate.

Tissyl chloride (13.1 ml) was added to 150 ml of benzyl alcohol, keeping the temperature below 0 ° (exothermic reaction). To students ^o raztrg> O was added dropwise to N- [1 (S) -carboxy-H-phenylpropyl] L- alanyl-L- overflow (15 g). The cooling bath was removed and the ace mixture was stirred at room temperature overnight. After heating to 45 'under vacuum to remove the dissolved gases, the reaction mixture was diluted with 500 ml of ether and washed with 10 x 100 ml of water. A solid phase appeared in the organic layer, which was filtered and dried to give 4.9 g of crude menobenzyl ester.

yao UHIGINAL

The combined aqueous extracts neutralized with sodium bicarbonate gave a second yield, weighing .4 g. The two yields were combined and one part (1.0 g) was recrystallized from ethanol-water to give 0.95 pure monobenzyl ester with m.p. 120 -125 °. Mass spectral analysis shows that the gasoline ester is attached to the proline ring.

Example 130. Ethyl N- [1 (S) -Egoxycarbonyl-3-phenylpropyl] alanyl-L-proline hydrochloride.

A solution of 3.0 g of N- [1 (S) -ethoxycarbonyl-3-phenylpropyl) -1-alanyl-L-proline in 50 ml of absolute ethanol was saturated with hydrogen chloride at 0 °. After staying overnight at 0 °, HC! and ethanol was removed in vacuo to give 3.1 g of an oily liquid which, after freeze-drying, gave a white solid. Thin layer chromatography and HPLC indicate only one component. The NMR spectrum shows two ethyl groups for the aromatic ring. The mass spectrum shows a molecular ion at 404 mp / e.

Example 131. N- (1-Ethoxycarbonyl-N- (2-chlorophenyl) propyl] 4. alanyl N-lrolin.

It's working. as described in Example 26, as 2-oxo-3- (2-chlorophenyl) butyrate and L-alanyl-L-proline were condensed in the presence of sodium cyanoborohydride to give N- (1-ethoxycarbonyl-3- (2-chlorophenyl) propyl] -1-alanyl-L-proline The mass spectrum shows one maximum at 411, (M ⁺ + 1) and a strong maximum at 392, (M-18).

Example 132. N-egoxycarbonyl N- (4-fluorophenyl) propyl] N- alanyl N-proline.

As described in Example 26, ethyl-2-oxo-4- (4-fluorophenyl) butyrate and L-alanyl-L-proline were condensed in the presence of sodium cyanoborohydride to give N- [1-ethoxycarbonyl-3- (4-fluorophenyl) propyl] -1-alanyl-L-proline. The mass spectrum shows a molecular ion at 394 and strong maxima at 376 (M-18, H ₂ O) and 375 (M19, F). The NMR spectrum is consistent with the proposed structure. An exploratory analysis shows one hydrate.

Example 133. N- [1-Ethoxycarbonyl-3- (2-naphthyl) propyl] -Balanyl-B-proline.

As described in Example 26, ethyl-2-oxo-4- (2-naphthyl) -butyrate and B-alanyl-B-proline were condensed in the presence of sodium cyanoborohydride to give N- [1-ethoxycarbonyl-3 - (2naphthyl) propyl] -B-alanyl-B-proline. The mass spectrum shows a molecular ion at 426 and a maximum at 408 (M-18, H ₂ O). The NMR spectrum is consistent with the proposed structure. The elemental analysis shows one hemihydrate.

Example 134. N- [1- [carboxy-3- (2-chlorophenyl) propyl] -blysyl-B-proline.

As described in Example 26, egyl-2-oxo-4- (2-chlorophenyl) butyrate and N-ε-tert.-Boc-B-lysyl-B-proline were condensed in the presence of sodium cyanoborohydride to give N-[alpha] - [1-carboxy-3- (2-chlorophenyl) propyl] - N-ε-tert-Boc-B-lysyl-B-proline. The tert-Boc-protecting group was removed by treatment with 4N HCl in ethyl acetate , then the ethyl ester group is saponified with dilute sodium hydroxide solution to give N-α- [1-carboxy-3- (2-chlorophenyl) propyl] -β-lysyl-β-proline after absorption on a highly acidic ion exchange resin and subsequent elution. Elemental analysis showed solvation with 1.5 mol of water. The NMR spectrum is consistent with the proposed structure. The mass spectrum of a silylated sample shows a maximum at 568, which corresponds to a mass loss of 15 (methyl) of the disilylated products.

Example 135. N- [alpha] - [1-carboxy-3- (4-fluorophenyl) propyl] -Blysyl-B-proline.

As described in Example 134, ethyl-2-oxo-4- (4fluorophenyl) butyrate and N-ε-tert.-Boc-B-lysyl-B-proline were condensed in the presence of sodium cyanoborohydride. The resulting product was unblocked as described in Example 4 to give N-α- [1carboxy-3- (4-fluorophenyl) propyl] -B-lysyl-B-proline. Elemental

Анализ · · Analysis shows sesquihydrate. The NMR spectrum is in accordance with the proposed structure. The mass spectrum of the silylated sample shows a molecular ion at 711 for the tetrasilylated products.

Example 136. N- [alpha] - [1-carboxy-3- (2-naphthyl) propyl] -1-lysyl-Lproline.

As described in Example 134, substitution of ethyl-2-oxo-4 (2-chlorophenyl) butyrate with ethyl 2-oxo-4- (2-naphthyl) butyrate gave N- [1-carboxy-3- ( 2-naphthyl) propyl] -1_-lysyl-L-proline. The NMR spectrum is compatible with the proposed structure. The mass spectrum shows a molecular ion at 599 for the distilled products.

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

A mixture of biphosphorus tribromide (18.95 g) and pyridine (3.76 ml) in 10 ml of benzene was cooled to -5 ° C and treated with a solution of 2- (2-thienyl) ethanol (25 g) in 10 ml of benzene containing 1.25 ml of pyridine at such a speed that the reaction temperature does not exceed 0 ° C. The mixture is then allowed to reach room temperature and stirred overnight. The reaction mixture was then concentrated in a water pump at 30 40 ° to remove the solvent and other volatile constituents, leaving the residue under Claisen distillation under vacuum from a water pump to obtain 21 g of crude bromide (b. 115 -119 ° C). This product was subjected to fractional distillation at 0.3 mm to give 19 g of pure 2- (2-thienyl) ethyl bromide (bp 50-51 ° C; 99.9%). This bromide was dissolved in 40 ml of dry tetrahydrofuran and added dropwise with stirring to a mixture of magnesium sawdust (2.4 g) and 15 ml of dry tetrahydrofuran in a nitrogen atmosphere. The mixture was then stirred and refluxed for 1 hour, cooled to room temperature and added dropwise under nitrogen to a mixture of diethyloxalate (17.5 g) in 25 ml of dry THF in an ice bath at -15 ° C. After completion of the reaction, the reaction mixture was stirred at -10 ° C for 15 minutes, 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 which was found by gas chromatography to contain, in wt%: solvent 26, diethyloxalate 10.5 and ethyl oxo- (2-thienyl) ) -bougrate 63.4. The NMR spectrum is consistent with this mixture, showing the presence of two specific types of ethyl esters, and the mass spectrum exhibits a maximum at m / e = 212 for the molecular ion of a ketoester.

Example 138. N- [1-Ethoxycarbonyl-3- (2-thienyl) propyl] -L alanyl-L-proline.

Add from type 4A molecular sieves (powders) to a 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 45 min at room temperature, followed by the slow addition (3.5 hours) of sodium cyanoborohydride solution (0.785 g) in 15 ml of anhydrous ethyl alcohol and stirring was continued overnight at room temperature. The reaction mixture was made slightly acidic with glacial acetic acid, filtered, and the filtrate was concentrated in vacuo to an oily orange liquid. Extraction of a chloroform solution of this oil with 10% potassium bicarbonate, followed by re-extraction of the acidified (pH 3) bicarbonate extracts with chloroform, after drying and evaporating the solvent, gave 400 mg of the desired disubstituted dipeptide. The NMR spectrum (CDCI3) shows the thienyl proton as a multiplet at 7.0 ppm, two active protons in a wide singlet at 6.2 ppm, six methyl protons as a wide multiplet of 1.22 - 1.43 ppm and one series of poorly defined multiplets in the range 1.8 - 4.4 ppm.

Example 139. N [1-Carboxy-3- (2-thienyl) propyl] 4.-lysyl-Lproline.

Type 4A molecular sieves were added to a mixture of Ν-ε-τρβτ.-Boc-L-lysyl-L-proline (2.96 g) and ethyl 2-oxo-4 (2-thienyl) -butyrate (6.4 g) in 40 ml of anhydrous ethyl alcohol. The mixture was stirred for 45 min at room temperature, then a solution of sodium cyanoborohydride (1.36 g) in 25 ml of anhydrous ethyl alcohol was added slowly (4.5 hours) and stirred overnight at room temperature. The reaction mixture was then acidified very slightly with glacial acetic acid, filtered and concentrated in vacuo to give an orange foam (16.4 g). After spraying this crude foam with three portions of ether and decanting the ether solution, a significant amount of excess ketoether was removed. The ether-insoluble oil was poured into 1.1 ml of cold ether, filtered and concentrated in vacuo to give an orange oily liquid. A solution of this unblocked product in 250 ml of ethyl alcohol was treated with 5 g of sodium hydroxide in 35 ml of water and stirred overnight at room temperature. After careful acidification with 0.5 N HCl, concentration in vacuo and subsequent purification by elution from one column of 150 ml of Dautex 50 W-X2 (H *) with water containing 2% pyridine, the substituted dipeptide was obtained in the form of dicarboxylic acid. The NMR spectrum (D ₂ O / DCI) is consistent with the texture.

Example 140 N- [1-hydroxy-34-enylpropyl] 4.-alanyl-amino-butyric acid.

A mixture of 8.9 g of methylaminoisobutyrate hydrochloride, 8.7 ml of triethylamine and 11.9 g of Boc-L-alanine in 135 ml of methylene chloride 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 46 hours, filtered and the filtrate was concentrated, then the residue was separated by treating the portions respectively with water and ethyl acetate. The organic phase was washed with 10% citric acid, water, 5% sodium bicarbonate and water. It is dried over magnesium sulfate and concentrated to give the crude dipeptide. Purify the crude product on silica gel, eluting with hexane-ethyl acetate (7: 3).

Dissolve 1.2 g of this dipeptide in 30 ml of 4N HCl / ethyl acetate and store for one hour at room temperature. Concentrate the mixture and dry the residue by freeze-drying to give the L-alanyl-α-amino butyric acid methyl ester hydrochloride

A solution of 688 mg of this amino ester, 949 mg of ethyl 2-oxo-4-phenylbutyrate and 328 mg of sodium acetate in 10 ml of ethanol is prepared. This mixture is hydrogenated over 0.5 g of 10% palladium on carbon at 276 kN / m ² . filtrate and concentrate the filtrate. Add 4 ml of 7: 3 hexane: ethyl acetate, filter, and chromatograph the filtrate on silica gel to give methyl N-ethoxycarbonyl-3-phenylpropyl] L-alanyl-α-aminoisobutyrate.

Elemental analysis: Calculated for (C 25 H 25 N 4 O): C, 63.30; H, 8.24; N, 7.38; found: C-63.58; H, 8.13; N, 7.27;

The NMR spectrum is compatible with the structure

Mix 460 mg of this diester with 3 ml of 2 N sodium hydroxide overnight at room temperature. Apply the resulting solution to a Dautex 50 (H *) column and elute with 5% pyridine in water. Concentrate the appropriate fractions to give N- (1-carboxy-3-phenylpropyl) -ALanyl-N-aminobutyric acid. Elemental analysis. Calcd for (C ₁₇ H ₂ 7N2O ₆ .2H ₂ O): C - 54.82; H, 7.58; Found: C, 54.94; H, 7.51; N, 7.48.

The NMR spectrum is compatible with the structure.

Example 141. b1 [1-Carboxy-3-phenylpropyl] -1'-alanyl-L · methylphenylalanine.

The compound was prepared according to Example 140 starting from the hydrochloride of 1-methyl-phenylalanine methyl ester instead of the hydrochloride of the amino-butyric acid methyl ester

Example 142. N {-1-carboxy-3- [4- (1-benzylimidazolyl)] propyl 1-alanyl-bproline.

Excess of 4- (4- (1-benzylylidazolyl)] - 2-ketobutyric acid (obtained by dibenzylation of 3- (4-imidazolyl) propionic acid, by condensation with diethyloxalate, followed by acid-catalyzed hydrolysis and decarboxylation) is condensed with Lalanyl-Lproline in water (pH 6.7) in the presence of excess sodium cyanoborohydride After 3 days at room temperature, the product is absorbed onto a highly acidic ion exchange resin After elution with 2% aqueous pyridine solution and freeze drying the crude product that follows is purified on a Sephadex LH-20 column using methanol C ¹³ and H ¹ NMR and IR spectral data are consistent with the structure.Organic elemental analysis data are available for the monohydrate The mass spectrum (after silylation) shows maxima at 572 (M * ), 577 (M * -15), and 455 (M ⁺ - CQSiMea) m / e.

Example 143. N [1-Carboxy-3- (4-imidazolyl) propyl] 4.-alanyl-L-proline.

N-1-carboxy-3-4- (1-benzimidazolyl) propyl-1-alanyl-Lproline prepared as described in Example 142 was subjected to hydrogenolysis at 276 kN / μ ² over an equal weight of 5% palladium on charcoal in a 3: 1 acetic acid / water mixture at 60 ° for three days. After removal of the catalyst by filtration and concentration of the filtrate, the crude product was purified by absorption on a highly acidic ion exchange resin, followed by elution with 2% aqueous pyridine and freeze drying. Further purification was performed on a Sephadex LH-20 column using methanol. H ¹ NMR and thin layer chromatography (silica gel) indicated that the gasoline group had been removed to obtain the title product of the example. The mass spectrum shows a maximum of (M ⁺ -18) at rn / e = 320.

Example 144. N-1-carboxy-3- [4- (1-benzylimidazolyl)] propyl} ^, -1-lysyl-bproline and

N [1-carboxy-3- (4-imidazolyl) propyl] -1-lysyl-b-proline.

The two title compounds of the example were prepared as N- -1-carboxy-3- [4- (1-benzylimidazolyl)] propyl N-85 alanyl-L-proline (Example 142) and N [1-carboxy-3 were obtained - (4-imidazolyl) propyl] L-alanyl-L · proline, replacing L-alanyl-L-proline with Et-Boc-Lysyl-1-proline and tert.-Boc group removed with trifluoroacetic acid at 0 ° for three hours after condensation

Example 145. N-1-ethoxycarbonyl-3- [4- (1-benzylimidazolyl)] propyl N-alanyl-1-proline and

N [1-ethoxycarbonyl-3- (4-imidazolyl) propyl] N-alanyl-L-proline.

The two compounds of the title of the example were prepared as N-1-carboxy-3- [4- (1-benzylimidazolyl)] propyl-C 1 alanylT-proline (Example 142) and N [1-carboxy-3- (4-imidazolyl) ) propyl] L-alanyl-L-proline (Example 143) using 4- [4 (1-benzylimidazolyl)] -2-ketobutyric acid ethyl ester and condensation was carried out in the presence of 4A sieve in ethyl alcohol, instead of in water.

Claims (10)

A pharmaceutical composition for the treatment of high blood pressure comprising a pharmaceutically acceptable carrier and a pharmaceutically effective amount of an amino acid derivative of the formula wherein R and R ⁸ are the same or different and mean hydroxy, lower alkoxy, lower alkeny, lower dialkylamino lower alkoxy, acylamino lower alkoxy, acyloxy lower alkoxy, aryloxy, aryl lower alkoxy, substituted aryloxy or substituted aryl lower alkoxy, the substituent being methyl, halogen or methoxy, amino, lower alkylamino, lower dialkylamino, aryl lower alkylamino or aryl lower alkylamino oxyamino, R ¹ is hydrogen, alkyl having from 1 to 20 carbon atoms, including side and cyclic and unsaturated alkyl groups; substituted lower alkyl, the substituent being halogen, hydroxy, lower alkoxy, aryloxy, amino, lower alkylamino, lower dialkylamino, acylamino, arylamino, guanidino, imidazolyl, indolyl, mercapto, lower alkylthiocarbothio, carboxylic, phenyl, substituted phenyl, the substituent being lower alkyl, lower alkoxy or halogen, aryl lower alkyl or heteroaryl lower alkyl, aryl lower alkenyl or heteroaryl lower alkenyl, substituted aryl lower alkenyl, substituted heteroaryl alkylarylalkyl; or lower alkenyl, the substituent being halogen or dihalogen, lower alkyl, hydroxy, lower alkoxy, amino, aminomethyl, tn acylamino, lower dialkylamino, lower alkylamino, carboxyl, halogenated lower alkyl, cyano or alkylsulfonylalkyl or alkylosulfonylalkyl substituted on the alkyl moiety by amino or benzoylamino; R ² and R ⁷ are hydrogen or lower alkyl; R ³ is hydrogen, lower alkyl, phenyl lower alkyl, aminomethyl phenyl lower alkyl, hydroxyphenyl lower alkyl, hydroxy lower alkyl, acetylamino lower alkyl acylamino lower alkyl amino lower alkyl, dimethylamino lower alkyl, halogenated lower alkyl, guanidino lower alkyl, imidazolyl lower alkyl, indolyl lower alkyl, mercapto lower alkyl, lower alkylthio lower alkyl; R ⁴ is hydrogen or lower alkyl; R ⁵ is hydrogen or lower alkyl, phenyl, phenyl lower alkyl, hydroxy lower alkyl, amino lower alkyl, guanidino lower alkyl, imidazolyl lower alkyl, indolyl lower alkyl, mercapto lower alkyl or lower alkyl; R ⁴ and R ⁵ may be interconnected to form an alkylene bridge of from 2 to 4 carbon atoms, an alkylene bridge of from 2 to 3 carbon atoms and one sulfur atom, an alkylene bridge ^having 3 to 4 carbon atoms containing one double bond or alkylene bridge, as indicated above, substituted by hydroxy, lower alkoxy, lower alkyl, or pharmaceutically acceptable salts thereof, and an antihypertensive and / or diuretic compound selected from the group consisting of amiloride, atenolol, bendroflumetazid, chloroflumetazid, chloro , chlorothiazide, clondin hydrochloride, cryptenamine acetate ati and kriptenamin tannate, deserpidine, diazoxide, guanethidine sulfate, hydralazine hydrochloride, hydrochlorothiazide, hydroflumethiazide, metolazone, metoprololtartarat, methyclothiazide, methyldopa, methyldopa hydrochloride, minoxidil, pargyline hydrochloride, polythiazide, prazosin hydrochloride, propranolol, rauvolfia coil retsinamin, reserpine, sodium nitroprusside, spironolactone, timolol, trichloromethacid, trimethophan camsylate, benzthiazide, quinetazone, ticrinaphane, triamterene, acetazolamide, aminophylline, cyclothiazide, ethacrynic acid, furosemide, meretox iline procaine, sodium ethacrynate and mixtures thereof and combinations thereof. 2. A pharmaceutical composition for treating high blood pressure comprising a pharmaceutically acceptable carrier and a pharmaceutical compound of an effective amount of an amino acid of formula B which R and R ⁶ may be independently and independently hydroxy, lower alkoxy, arylalkyloxy, R ² and R ⁷ are hydrogen, R ³ is methyl, amino lower alkyl, R ⁴ and R ⁵ are bonded via a carbon and nitrogen atom, for form proline, 4-thiaprolin or 4-methoxyproline and R ¹ is alkyl having from 1 to 8 carbon atoms; substituted lower alkyl, where the alkyl group has 1 to 5 carbon atoms and the substituent is amino, arylthio or aryloxy, arylalkyl or heteroarylalkyl, wherein the alkyl moiety has 1 to 3 carbon atoms, substituted arylalkyl or heteroarylalkyl, such as the alkyl group having 1 and 5 carbon atoms and the substituent (s) is halogen? dichalogen amino, aminoalkyl, hydroxy, lower alkoxy or lower alkyl, its pharmaceutically acceptable salts, and an antihypertensive and / or diuretic compound selected from the group comprising amiloride, atenolol, bendroflumetidine, chlorothalidine, chlorothalidine, chlorothalidon, chlorothalidine, chlorothalidon deserpidine, diazoxide, quanethidine sulfate, hydralazine hydrochloride, hydrochlorothiazide, hydroflumethiazide, metolazone, metoprolaltartarate, methylcothiazide, methyldopa, methyldopate hydrochloride, minoxide, pargillin hydrochloride iazid, prazosin hydrochloride, propranolol, rauvolfia coil restsinamin, reserpine, sodium nitroprusside, spironolactone, timolol, trichlormethyacid, trimetofan camsylate, benzthiazide, quinethazone, tikrinafan, triamterene, acetazolamide, aminophylline, cyclothiazide, ethacrynic acid, furosemide, procaine meretoksilin. ethacrynate sodium and mixtures thereof and combinations thereof. The composition of claim 2, wherein the amino acid compound is selected from the group consisting of: N- (1-carboxy-3-phenylpropyl) -1-alanyl-1-proline; N- (1-ethoxycarbonyl-3-phenylpropyl) -1.-alanyl-1-proline N- (1-ethoxycarbonyl-4-methylpentyl) -1-alanyl-1-proline; N- (1-carboxy-5-aminopentyl) -1_-alanyl-1-proline; N- [alpha] - (1-carboxy-34 © nylpropyl) -Lysyl-L-proline; N- [alpha] - (1-ethoxycarbonyl-3-phenylpropyl) -1-lysyl-bproline; N- [alpha] - [1-carboxy-3- (3-indolyl) propyl] N-lysyl-L-proline; N-α- [1-carboxy-3- (4-chlorophenyl) propyl] N-lysyl-L-proline; N- [alpha] - [1-carboxy-2-phenylthioethyl] -1-lysyl-L-proline; N- [alpha] - [1-carboxy-3- (4-chlorophenyl) propyl] -E-lysyl-trans-4methoxy-E-proline; N- [alpha] - [1-carboxy-5-aminopentyl] N-lysyl-L-proline; N- [alpha] - [1-carboxy-34-enylpropyl] -E-ornyl-L-proline; Ethyl N- (1-ethoxycarbonyl-3-phenylpropyl N-alanyl-E-proline hydrochloride; N- [1- (ethoxycarbonyl) -3- (4-imidazolyl) propyl) -1-alanine <proline; N- (1 (C) -carboxy-3-phenylpropyl) -E-alanyl-E-proline; N- (1 (c) -ethoxycarbonyl-3-phenylpropyl) -E-alanyl-E-proline; N- (1 (S) -ethoxycarbonyl-3-phenylpropyl) -L-alanyl-E-proline, maleic salt; N-a- (1 (S) -carboxy-3-phenylpropyl) 4.-lysyl-L-proline; Ethyl N- (1 (c) -ethoxycarbonyl-3-phenylpropyl) -L-alanyl-L · proline hydrochloride; and N-α- (1 (c) -ethoxycarbonyl-8H () naphthyl) -E-lysyl-L-proline; A pharmaceutical composition for treating high blood pressure comprising a pharmaceutically effective amount of an amino acid compound according to claim 3, a pharmaceutically effective amount of hydrochlorothiazide and a pharmaceutically acceptable carrier. A pharmaceutical composition for treating high blood pressure comprising a pharmaceutically effective amount of an amino acid compound according to claim 3, a pharmaceutically effective amount of hydrochlorothiazide and timolol, and a pharmaceutically acceptable carrier. A pharmaceutical composition for treating high blood pressure comprising a pharmaceutically effective amount of an amino acid compound according to claim 3, a pharmaceutically effective amount of timolol and a pharmaceutically acceptable carrier. A pharmaceutical composition for treating high blood pressure comprising a pharmaceutically effective amount of an amino acid compound according to claim 3, a pharmaceutically effective amount of hydrochlorothiazide and amiloride, and a pharmaceutically acceptable carrier. A pharmaceutical composition for treating high blood pressure comprising a pharmaceutically effective amount of an amino acid compound according to claim 3, a pharmaceutically effective amount of furosemide, and a pharmaceutically acceptable carrier. 9. A pharmaceutical composition for treating high blood pressure comprising a pharmaceutically effective amount of N- (1 (S) ethoxycarbonyl-3-phenylpropyl) -1_-alanyl-1-proline, a pharmaceutically effective amount of hydrochlorothiazide and a pharmaceutically acceptable carrier. A pharmaceutical composition for treating high blood pressure comprising a pharmaceutically effective amount of N-α- (1 (S) carboxy-3-phenylpropyl) -1-lysyl-L-proline, a pharmaceutically effective amount of hydrochlorothiazide and a pharmaceutically acceptable carrier. A pharmaceutical composition for the treatment of high blood pressure comprising a pharmaceutically effective amount of N- (1 (S) ethoxycarbonyl-3-phenylpropyl) -L-alanyl-1-proline, a pharmaceutically effective amount of hydrochlorothiazide and timolol and a pharmaceutically acceptable carrier. 12. A pharmaceutical composition for treating high blood pressure comprising a pharmaceutically effective amount of Na- (1 (S) carboxy-3-phenylpropyl) -1H-lysyl-L-proline, a pharmaceutically effective amount of hydrochlorothiazide and timolol and a pharmaceutically acceptable carrier . A pharmaceutical composition for the treatment of high blood pressure comprising a pharmaceutically effective amount of N- (1 (S) ethoxycarbonyl-3-phenylpropyl) -E-alanyl-k-proline, a pharmaceutically effective amount of timolol and a pharmaceutically acceptable carrier. 14. A pharmaceutical composition for treating high blood pressure comprising a pharmaceutically effective amount of N-? - (1 (S) carboxy-3-phenylpropyl) -1_-lysyl-L-proline, a pharmaceutically effective amount of timolol and a pharmaceutically acceptable carrier. 15. A pharmaceutical composition for treating high blood pressure comprising a pharmaceutically effective amount of N- (1 (S) ethoxycarbonyl-3-phenylpropyl) -1-alanyl-E-proline, a pharmaceutically effective amount of hydrochlorothiazide and amiloride and a pharmaceutically acceptable carrier. 16. A pharmaceutical composition for treating high blood pressure comprising a pharmaceutically effective amount of N-α- (1 (S) carboxy-3-phenylpropyl) -1-lysyl-L-proline, a pharmaceutically effective amount of hydrochlorothiazide and amiloride and a pharmaceutically acceptable carrier. 17. A pharmaceutical composition for treating high blood pressure comprising a pharmaceutically effective amount of N- (1 (S) ethoxycarbonyl-3-phenylpropyl) -1_-alanyl-1-proline, a pharmaceutically effective amount of furosemide and a pharmaceutically acceptable carrier. 18. A pharmaceutical composition for treating high blood pressure comprising a pharmaceutically effective amount of N-? - (1 (S) carboxy-3-phenylpropyl) -Lysyl-L-proline, a pharmaceutically effective amount of furosemide and a pharmaceutically acceptable carrier. 19. A process for the preparation of a compound of the formula wherein R and R ⁶ are the same or different and denote hydroxy, lower alkoxy, lower alkenoxy, lower dialkylamino lower alkoxy, acylamino lower alkoxy, acyloxy lower alkoxy, aryloxy, aryl lower alkoxy, instead or substituted aryl lower alkoxy, the substituent being methyl, halogen or methoxy, amino, lower alkylamino, lower dialkylamino, aryl lower alkylamino or hydroxyamino, R ² and R ⁷ are hydrogen or lower alkyl; R ³ is hydrogen, lower alkil.fenil lower alkyl, aminomethyl phenyl lower alkyl, hydroxyphenyl lower alkyl, hydroxy lower alkyl, acetylamino lower alkyl acylamino lower alkyl amino lower alkyl, dimethylamino lower alkyl, halogenated lower alkyl, guanidino lower alkyl, imidazolyl lower alkyl, indolyl lower alkyl, mercapto lower alkyl, lower alkylthio lower alkyl; R ¹ is hydrogen, alkyl having from 1 to 20 carbon atoms, including side and cyclic and unsaturated alkyl groups; substituted lower alkyl, the substituent being halogen, hydroxy, lower alkoxy, aryloxy, amino, lower alkylamino, lower dialkylamino, acylamino, arylamino, guanidino, imidazolyl, indolyl, mercapto, lower alkylthiocarbothio, carboxylic, phenyl, substituted phenyl, the substituent being lower alkyl, lower alkoxy or halogen, aryl lower alkyl or heteroaryl lower alkyl, aryl lower alkenyl or heteroaryl lower alkenyl, substituted aryl lower alkenyl, substituted heteroaryl alkylarylalkyl; or lower alkenyl, the substituent being halogen or dihalogen, lower alkyl, hydroxy, lower alkoxy, amino, aminomethyl, acylamino, lower dialkylamino, lower alkylamino, carboxyl, halogenated lower alkyl, cyano or alkylsulfonylalkyl, or alkylsulfonylalkyl in the alkyl moiety with amino or acylamino; R ⁴ is hydrogen or lower alkyl; R ⁵ is hydrogen or lower alkyl, phenyl, phenyl lower alkyl, hydroxy lower alkyl, amino lower alkyl, guanidino lower alkyl, imidazolyl lower alkyl, indolyl lower alkyl, mercapto lower alkyl or lower alkyl; R ⁴ and R ⁵ . are. linked together to form an alkylene bridge of 2 to 4 carbon atoms, an alkylene bridge of 2 to 3 carbon atoms and one sulfur atom, an alkylene bridge of 3 to 4 carbon atoms containing a single double bond or an alkylene bridge, as indicated » above substituted by hydroxy, lower alkoxy or lower alkyl, and pharmaceutically acceptable salts thereof, wherein a ketone of the formula in which R ¹ includes a suitable protection of each reagent group is reacted with a dipeptide or a protective dipeptide of formula r · ·· In which R ³ and R ⁶ include appropriate protection of each reagent? Group and the intermediates formed are reduced by the reaction in the presence of a reducing agent, followed by removal of the protecting groups, if necessary to obtain the desired product, and if desired its salt is obtained by conventional methods and optionally isolated biologically active isomer by chromatography or fractional crystallization. A process for preparing a compound according to claim comprising reacting a ketone with the formula ΠΟ19. in which R is a reactive group, the formula is not hydroxy but R is a suitable protection of each with an amino acid or a protected amino acid with R? HjN-COOH R ³ includes adequate protection of each reactive. a group, in the presence of a reducing agent, obtains an intermediate of formula (I) in which an agent to, even after amino acid of formula n-coon, bind this intermediate or protected amino acid derivative (VI) LhX ^ ooh and condensation of a compound of formula (X) with an amino acid derivative of formula R ⁴ R 5 (VI) wherein R ⁶ is not hydroxy and R ⁵ includes appropriate protection of each reactive group, followed by deprotection if necessary to obtain a compound according to claim 18, wherein R and R ⁶ are not hydroxy and, if desired, R and / or R * to be converted to hydroxyl groups by hydrolyzing or hydrogenating the corresponding precursor and further, if desired, preparing a salt thereof by conventional methods and, if desired, further a biologically more active isomer is isolated by chromatography or fractional crystallization. A method for the preparation of compounds according to claim 19, comprising reacting a dipeptide of the formula reactive (»0 in which R ³ and R ⁵ include appropriate protection of each group with a compound of the formula R 2 (I) wherein R ⁶ is not hydroxy and R ⁵ includes a suitable protection of each reagent group to give a compound according to claim 19 wherein R and R ⁶ are not hydroxy, followed by removal of the protecting groups at optionally converting R and / or R ⁶ into hydroxy by hydrolyzing or hydrogenating the corresponding intermediate, and optionally preparing a salt by conventional means and, if desired, isolating the biologically active isomer by chromatography or fractional crystallization. 21. A process for the preparation of a compound of formula according to claim 19, which comprises reacting an amine of formula reactive which is a R ¹ group. wherein R ³ and includes with ketone with appropriate protection of each formula O = -C ~ c - N - N - CO - R ⁸ (VIII) R ⁵ include appropriate protection of each reactive group in the presence of a reducing agent, followed by deprotection if necessary to obtain the desired product or, if desired, a stepwise reaction by condensation of (VII), wherein R 5 is hydroxy with a keto acid of formula wherein R ³ includes appropriate protection of each reactive group in the presence of a reducing agent to give a compound of formula wherein R ¹ includes appropriate protection of each reactive group and X is chlorine, bromine, iodine or a sulfonyloxy derivative, sequ ano by removal of the protecting groups if necessary to yield the desired product or if desired reacting (XI), in which R is not OH with an amino acid derivative of the formula Su-UN wherein R ³ comprises a suitable protection of each reactive group to form an intermediate of formula and then the intermediate is reacted with an amino acid derivative of formula -CO-R ⁸ in which R ⁶ is not hydroxy, and R ⁵ includes a suitable protection of each reactive group, followed by removal of the protecting group, if necessary to obtain a compound according to claim 19, and optionally to obtain its salt by conventional methods and, if desired, isolate the biologically more active isomer by chromatography or fractional crystallization. 23. A process for the preparation of compounds according to claim 19, wherein said amino acid comprises reacting the derivative of the formula IN WHICH the reactive amino acid includes appropriate protection of any group with an α-substituted acyl derivative in which X is a chlorine, bromine, iodine or sulfonyloxy derivative, in which preferably R® is not hydroxy and R ³ and R ⁵ include appropriate protection of each a reactive group, followed by removal of the protecting group if necessary. to obtain the desired product or, optionally, the amino acid ester of formula (VII) in which R is not hydroxy, is reacted with an α-substituted acid of formula R? X-N-H-COOH in which R ³ includes a suitable protection of each reactive group to obtain an intermediate ester of formula and reacting an amino acid of the formula intermediate with an ester in which R ⁶ is not hydroxy, and R ⁵ includes a suitable protecting each reactive group, followed by removal of the protecting group, if necessary, to obtain a compound according to claim 19, and optionally converting R and / or R® into hydroxy by hydrolyzing or hydrogenating the corresponding precursor and further, if desired, its salt is obtained by co conventional methods and, if desired, the biologically more active isomer is isolated by chromatography or fractional crystallization. A method according to claim 19 for the preparation of a compound of formula R and R can independently and independently be hydroxy, alkoxy, arylalkyloxy, R ¹ is alkyl having from 1 to 8 carbon atoms; substituted lower alkyl, wherein the alkyl group has 1 to 5 carbon atoms and the substituent is amino, arylthio or aryloxy, arylalkyl or heteroarylalkyl, wherein the alkyl moiety has 1 to 3 carbon atoms, a substituted arylalkyl or heteroarylalkyl group, such as the alkydo group having 1 to 3 carbon atoms the substituent (s) is halogen dihalogener amino, aminoalkyl, hydroxy, lower alkoxy or lower alkyl, R ² and R ⁷ are hydrogen, R ³ is methyl, amino lower alkyl, R ⁴ and R ⁵ are bonded via a carbon and nitrogen atom, for to form proline, 4-thiaprolin or 4methoxyproline. The method of claim 19 for the preparation of a compound of formula 100 comprising reacting the ketone of the formula with the dipeptide of the formula in the presence of a reducing agent to obtain the desired product, and isolating the more active diastereoisomer by chromatography and fractional crystallization. 26. The method of claim 25, wherein the maleic acid of N- [1 (S) ethoxycarbonyl-3-phenylpropyl] -alanilyl-bproline is obtained by fractional crystallization. A method according to claim 19 for the preparation of a compound of formula NHj nnr n which involves the reaction of a ketone of formula 101 O CHpHiD but-with-ο-ο with a protective dipeptide of the formula in which t-Boc is the t-butyloxycarbonyl protecting group in the presence of a reducing agent, to obtain the protected form of the desired product, is followed by reaction with an appropriate acid reagent, to obtain the desired product, followed by isolation of the biologically more active diastereomer by chromatography or fractional crystallization. A method according to claim 19 for the preparation of a compound of formula which comprises the reaction of a ketone of formula <feHs C2H5O-c-CH = -O 102 with a protected dipeptide of formula NhftBoc (CHL in which t-Boc is the t-butyloxycarbonyl protecting group, in the presence of a reducing agent, to obtain the protected form of the desired product, is followed by reaction with a suitable acid reagent to obtain the desired product, followed by isolation of the biologically more active diastereomer by chromatography or fractional crystallization. A method according to claim 19 for the preparation of a compound of formula comprising the reaction of a ketone of formula with a protected dipeptide of formula 103 M-HBoc in which t-Boc is the t-butyloxycarbonyl protecting group, in the presence of a reducing agent, to obtain the protected form of the desired product, followed by reaction with a suitable acid reagent to obtain the desired product, and isolation of the biologically more active diastereomer by chromatography or fractional crystallization. 30. The method of claim 19 for the preparation of the following compounds: N- (1-carboxy-3-phenylpropyl) -1_-alanyl-bproline; N- (1-ethoxycarbonyl-3-phenylpropyl) -E-alanyl-L-proline N- (1-ethoxycarbonyl-4-methylpentyl) -1_-alanyl-1-proline; N- (1-carboxy-5-aminopentyl) -1_-alanyl-bproline; N- [alpha] - (1-carboxy-34enylpropyl) N-lysyl-L-proline; N- [alpha] - (1-ethoxycarbonyl-3-phenylpropyl) -E-lysyl-proline; N-α- [1-carboxy-3- (3-indolyl) propyl] 4-lysyl-L-proline; N-α- [1-carboxy-3- (4-chlorophenyl) propyl] N-lysyl-L-proline; N- [alpha] - [1-carboxy-2-phenylthioethyl] -Lysyl-L-proline; N-α- [1-carboxy-3- (4-chlorophenyl) propyl] -1-lysyl-trans-4-methoxy-proline; N- [alpha] - [1-carboxy-5-aminopentyl] -1-lysyl-L-proline; N- [alpha] - [1-carboxy-3-phenylpropyl] N-ornigyl-L-proline; Ethyl N- (1-ethoxycarbonyl-3-phenylpropyl) N-alanyl-L-proline hydrochloride; N- [1- (ethoxycarbonyl) -3- (4-imidazolyl) propyl) -E-alanine-Aproline; N- [1-carboxy-3- (4-imidazolyl) propyl] N-lysyl-L-proline; 104 N- (1 (S) -carboxy-3-phenylpropyl) -alanil-E-proline; N- (1 (c) -ethoxycarbonyl-3-phenylpropyl) -1-alanyl-1-proline; N- (1 (S) -ethoxycarbonyl-3-phenylpropyl) -1-alanyl-E-proline, maleic salt; N-a- (1 (S) -carboxy-3-phenylpropyl) -lysyl-L-proline and Ethyl N- (1 (S) -ethoxycarbonyl-3-phenylpropyl) -alanilyl-L proline hydrochloride; and N-α- (1 (S) -ethoxycarbonyl-3-phenylpropyl) -1-lysyl-L-proline; 31. A pharmaceutical preparation for the treatment of high blood pressure comprising a pharmaceutically acceptable carrier and a pharmaceutically effective amount of an amino acid compound of the formula wherein R and R * are the same or different and mean hydroxy, lower alkoxy, lower alkeny, lower dialkylamino lower alkoxy, acylamino lower alkoxy, acyloxy lower alkoxy, aryloxy, aryl lower alkoxy, substituted aryloxy or substituted aryl lower alkoxy, the substituent being methyl, halogen or methoxy, amino, lower alkylamino, lower dialkylamino, aryl or lower alkyl hydroxyamino, R ¹ is hydrogen, alkyl having from 1 to 20 carbon atoms, including side and cyclic and unsaturated alkyl groups; substituted lower alkyl, the substituent being halogen, hydroxy, lower alkoxy, aryloxy, amino, lower alkylamino, lower dialkylamino, acylamino, arylamino, guanidino, imidazolyl, indolyl, mercapto, lower alkylthiocarboxyl, carboxy, alkoxy, or alkoxy substituted phenyl, the substituent being lower alkyl, lower alkoxy or halogen, aryl lower alkyl or heteroaryl lower alkyl, aryl lower alkenyl or heteroaryl lower alkenyl, substituted aryl lower alkyl, 105 substituted heteroaryl lower alkyl, substituted aryl lower alkenyl or substituted heteroaryl lower alkenyl, the substituent being halogen or dihalogen, lower alkyl, hydroxy, lower alkoxy, amino, aminomethyl, acylamino, lower dialkylalkylaminoalkylaminoalkylaminoalkylaminoalkylaminoalkylaminoalkylaminoalkylaminoalkylaminoalkylaminoalkylaminoalkylaminoalkylaminoalkylaminoalkylaminoalkylaminoalkylaminoalkylaminoalkylaminoalkylaminoalkylaminoalkylaminoalkylaminoalkylaminoalkylaminoalkylaminoalkylaminoalkylaminoalkylaminoalkylamino or sulfonamido, aryl lower alkyl or heteroaryl lower alkyl substituted in the alkyl moiety by amino or benzoylamino; R ² and R ⁷ are hydrogen or lower alkyl; R ³ is hydrogen, lower alkyl, phenyl lower alkyl, aminomethyl phenyl lower alkyl, hydroxyphenyl lower alkyl, hydroxy lower alkyl, acetylamino lower alkyl acylamino lower alkyl amino lower alkyl, dimethylamino lower alkyl, halogenated lower alkyl, guanidino lower alkyl, imidazolyl lower alkyl, indolyl lower alkyl, mercapto lower alkyl, lower alkylthio lower alkyl; R ⁴ is hydrogen or lower alkyl; R ⁵ is hydrogen or lower alkyl, phenyl, phenyl lower alkyl, hydroxy lower alkyl, amino lower alkyl, guanidino lower alkyl, imidazolyl lower alkyl, indolyl lower alkyl, mercapto lower alkyl or lower alkyl; R ⁴ and R ⁵ may be linked together to form an alkylene bridge of 2 to 4 carbon atoms, an alkylene bridge of 2 to 3 carbon atoms and one sulfur atom, an alkylene bridge ^& 3 to 4 carbon atoms containing one double bond or alkylene bridge as above substituted with hydroxy, lower alkoxy, lower alkyl or their pharmaceutically acceptable salts and an antihypertensive and / or diuretic compound and mixtures or combinations thereof. 106 32. A pharmaceutical composition for the treatment of high blood pressure, vklyuchvdsht pharmaceutically acceptable carrier and a pharmaceutically effective amount of compound of the amino acids of formula wherein R and R ⁶ each independently may baddt hydroxy, lower alkoxy, arylalkyloxy, R ² and R ⁷ are hydrogen, R ⁵ is methyl, amino lower alkyl, R ^ and R ⁵ are joined by carbon atoms and nitrogen to 00PA3YBAT proline, 4-thiaproline or 4-methoxy-proline and R ¹ is alkyl having from 1 to 8 CARBON ATOMS SUBSTITUTED LOWER ALKYL AS THE ALKYL GROUP HAS 1 TO 5 CARBON ATOMS AND SUBSTITUTES Amen, arylthio or aryloxy, arylalkyl or heteroarylalkyl wherein the alkyl moiety has from 1 to 3 carbon atoms, substituted arylalkyl or heteroalkyl, such as alkyl group having from 1 to 3 carbon atoms and substituent (s) is halo, dihalo, amino, aminoalkyl , hydroxy, lower alkoxy or lower alkyl or pharmaceutically acceptable salts thereof and an antihypertensive and / or diuretic compound and mixtures thereof, or mixtures thereof. The composition of claim 32, wherein the amino acid compound is selected from the following compounds: N- (1-carboxy-3-phenylpropyl) -1_-alanyl-1-proline; N- (1-ethoxycarbonyl-3-phenylpropyl) -1_-alanyl-bproline N- (1-ethoxycarbonyl-4-methylpentyl) -alanil-1-proline; 10? N- (1-carboxy-5-aminopentyl) -k-alanyl-L-proline; N- [alpha] (1-carboxy-3-phenylpropyl) -Lysyl-L-proline; N- [alpha] - (1-ethoxycarbonyl-3-phenylpropyl) -E-lysyl-k-proline; N-α- [1-carboxy-3- (3-indolyl) propyl] -1-lysyl-L-proline; N-α- [1-carboxy-3- (4-chlorophenyl) propyl] -1-lysyl-L-proline; N- [alpha] - [1-carboxy-2-phenyphyoethyl] -E-lysyl-L-proline; N- [alpha] - [1-carboxy-3- (4-chlorophenyl) propyl] -1-isyl-trans-4-methoxy-L-proline; N- [alpha] - [1-carboxy-5-aminopontyl] -k-lysyl-L-proline; N- [alpha] - [1-carboxy-3-phenylpropyl] 4.-ornyl-L-proline; Ethyl N- (1-ethoxycarbonyl-3-phenylpropyl) -1-alanyl-k-proline hydrochloride; N- [1- (ethoxycarbonyl) -3- (4-imidazolyl) propyl) -k-alanine-Aproline; N- [1-carboxy-3- (4-imidazolyl) propyl] -k-lysyl-L-proline; LC1 (S) -carboxy-3-phenylpropyl) -L-alanyl-k-proline; N- (1 (c) -ethoxycarbonyl-3-phenylpropyl) -k-alanyl-k-proline; N- (1 (S) -ethoxycarbonyl-3-phenylpropyl) -E-alanyl-k-proline, maleic salt; N- [alpha] - (1 (S) -carboxy-3-phenylpropyl) -k-lysyl-L-proline and Ethyl N- (1 (c) -ethoxycarbonyl-3-phenylpropyl) -k-alanyl-quinoline hydrochloride; and N-α- (1 (S) -btoxycarbonyl-3-phenylpropyl) -L-lysyl-L-proline.