KR840002357B1 - Process for the preparation of s-(acylamido acyl)mercapto acyl prolines - Google Patents

Abstract

Title compds. of formula (I), which are used as antihypertensives, are prepd. by reacting compds. of formula (II) with R2-H to give compds. of formula (III); removing R3 from (III) to form compds. of formula (IV); and reacting (IV) with R-A-0H. In the formulas, R is H, formyl, acetyl, propanoyl, butanoyl, phenylacetyl, phenylpropanoyl, benzoyl, cyclopentylcarbonyl, etc.; A is L-phenylalanyl, glycyl, L- alanyl, L-tryptophyl, L-tyrosyl, Lisoleucyl, etc.; R1 is H or methyl; R2 is L-proline, L-3,4-dehydroproline, D,L-3,4-dehydroproline etc.; R3 is acetyl; and n is 0 or 1, such that when n=0, R1 is methyl.

Description

Manufacturing method of S- (acyl amido acyl) mercapto acyl proline

Angiotensin converting enzyme (peptidyldipeptide hydrolase, hereafter called ACE) plays an important role in hypertension physiology.

The enzyme can convert angiotensin I, a decapeptide having the sequence AspArgValTyrIleHisProPheHisLeu, to an octapeptide angiotelsin II by removing the HisLeu at the carboxyl terminus.

The symbols for the various chemicals are described in the following table.

Ala = Left-Alanine

Arg = Left-arginine

Asp = L-aspartic acid

Gllu = pyro-Lglutamic acid

Gly = glycine

Hip = Hippuric acid (Benzoyl-glycine)

His = L-histidine

Ile = Left-Isoleucine

Leu = L-leucine

Phe = L-phenylalanine

Pro = Left-Proline

Pro = Left-3, 4-dehydroproline (L-3,4-dehydroproline)

Ser = L-serine

Trp = Left-tryptophan

Tyr = L-tryrosine

Val = L-valine

ACE = Angiotensin converting enzyme

Hopes = N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid.

Angiotensin I is found in kidneys and other tissues and plasma and is formed by the activity of renin, a peptidase that acts on α-2 globulin in serum.

Blood pressure is affected by certain peptides found in the blood, one of which is angiotensin II, a potent blood pressure booster.

Another is bradykinin, a nonapeptide with the arrangement ArgProProGlyPheSerProp heArg, a potent blood pressure lowering agent. In addition to direct blood pressure-boosting effects, angiotensin II stimulates the release of aldosterone, which tends to raise blood pressure by retaining extracellular salt and water.

Angiotensin II is found in measurable amounts in normal human blood but highly concentrated in blood of kidney hypertensive patients.

ACE activity levels are normal or hypertensive, usually exceeding the amount needed to maintain the measured level of angiotensin II.

However, it has been shown that significant antihypertensive effects can be achieved by prescribing ACE inhibitors to hypertensive patients. [Gavras, I., et al., New Engl. J. Med. 291, 817 (1974)]

The present invention relates to a method for preparing a new ACE inhibitor having the following general formula.

Wherein R is hydrogen, formyl, acetyl, propane oil, butane oil, waste acetyl, phenyl propane oil, benzoyl, cyclopentylcarbonyl, tert-butyloxycarbonyl, cyclopentylcarbonyl-L-lysyl, pyro-L -Gluramyl-L-lysyl, L-lysyl, L-arginyl or pyro-L-glutamyl,

A is L-phenylalanyl, glycyl, L-alanyl, L-tryptiphyl, L-tyrosyl, L-isosolyl, L-leusil, L-histidyl, or L-valyl and The amino group is amide-bonded with R under the condition that when phenyl is benzoyl, phenylalanyl is racemic.

R ₁ is hydrogen or methyl,

와 이미드 결합하고 있다.R ₂ is L-proline, L-3, 4-dehydrate proline, D, L-3, 4-dehydrate proline, L-3-hydroxyproline, L-4-hydroxyproline or L-thiazolidine-4 -Carboxylic acids and their amino groups are adjacent

And imide bonds.

n is 0 or 1 and when n = 0 R ₁ is methyl.

All amino acids mentioned herein have a left-coordinating configuration, unless otherwise specified, but in this benzoyl phenylalanine is racemic acid.

The compounds described are ACE inhibitors and they are useful as effective antihypertensive agents in oral administration.

Discovering the efficacy of ACE inhibitors in the compounds prepared by the present invention has opened the way for the synthesis of inhibitory compounds.

Although many inhibitors in the prior art were proline derivatives, using proline versus other amino acids also yielded good inhibitors. Arginine, phenylalanine, and alanine are all effective proline substitutes, so no certain trend can be recognized.

Substitutions for proline with L-3, 4-dehydrate proline have been made in several systems. Substitution of L-3 and 4-ΔPro at the seventh position of the bradykinin produced a bradykinin derivative, which significantly reduced the physiological activity. Fisher, GH et al., Arch. Biochem. Biophys, 189, 81 (1978). On the other hand, substitution of L-3, 4-ΔPro at the 3, 5 or 9 positions of the ACE inhibitor BPP _9a increases their inhibitory activity.

At present, however, no rational rationale has been established to explain the various results observed after the substitution of proline with ΔPro, and there is no clear concept of the effect of other proline derivatives or analogs substituted at various loci on ACE inhibitors. .

To date, the role of the left amino acid of sulfur in the structural formulas shown above is not correct and it is believed that this amino acid acts as an additional site for the enzyme.

If this is true, compounds with amino acids are expected to be better inhibitors.

Even so, it is not known which amino acids are effective at this position and enhance the inhibitory action of a given compound. Applicants believe that several amino acids are effective and that hydroxyproline, proline, L-, D, L-3, 4-desuproline and riazolidine-4-carboxylic acid derivatives are all effective antihypertensive agents and have high ACE inhibitory effects. I found out that I have

The present invention will be described in more detail in the following examples. Among these examples, thin layer chromatography (TLC) was performed using a silica gel plate, and the solvent system required for the TLC process is as follows.

(1) is methanol: chloroform of 1: 1 (volume ratio)

(2) Benzene: Water: Acetic Acid 9: 1: 9 (volume ratio)

(3) is acetic acid: water: n-butanol 26: 24: 150 (volume ratio)

(4) n-butanol: pyridine: acetic acid: water 15: 10: 3: 12 (volume ratio)

(5) Chloroform: Methanol: Ammonium hydroxide 60: 45: 20 (volume ratio)

Buffer for paper electrophoresis at pH 1.9: formic acid: acetic acid: water ratio 3: 2: 25 (volume ratio); pH 5.0: diethylene glycol: acetic acid: phytidine: water ratio 100: 6: 8.5: 885 ( Volume ratio).

Derivatives of amino acids such as tert-butyloxycarbonyl, benzoyl, acetyl, formyl, propane oil, butan oil, phenylacetyl, and phenyl propane oil are commercially available and are available from Sephadex LH-20, Sephadex G-10 and Sephadex G. -25 is a registered trademark of Pharmacia, Inc., in Uppsala, Sweden.

Example 1

ACE activity analysis

2×10^-4M)인 벤조일-글리신히스티딘루우신과 약 130,000cpm의 [3H] 벤조일글리신히스티딘루우실(25Ci/mmole)이었다. 효소를 앞의 완충제에 희석시켰더니 40μl의 완충된 효소는 섭씨 37도에서 항온처리 15분이내에 기질의 13%를 가수분해할 수 있었다.In most of the experiments described herein, enzymes were analyzed using 0.05 M Hepes buffer, pH 8, containing 0.1 M NaCl and 0.75 M Na ₂ SO ₄ . The substrate used has a final concentration of 1 × 10 ^-4 M, (K _m

2 × 10 ⁻⁴ M) benzoyl-glycine histidine leucine and about 130,000 cpm [3H] benzoylglycine histidine leusil (25 Ci / mmole). When the enzyme was diluted in the previous buffer, 40 μl of the buffered enzyme was able to hydrolyze 13% of the substrate within 15 minutes of incubation at 37 degrees Celsius.

To begin the assay, 40 μl of enzyme and 10 μl of water, or inhibitors dissolved in water, were preincubated for 5 minutes at 37 degrees Celsius. Then 50 μl of substrate was added to initiate the reaction and the solution was incubated for 15 minutes at 37 degrees Celsius.

To terminate this reaction, 1 ml of 0.1 M HCl was added followed by 1 ml of ethyl acetate.

The mixture was shaken with a rotary mixer and the phases separated by simple centrifugation. An aliquot of 500 μl of the ethyl acetate layer was transferred to a liquid scintillation vial containing 10 ml of Riafluor (registered trademark of New England Nuclear Company in Boston, Mass.), And then the enzyme concentration was constant to determine the I ₅₀ value. With changes, the activity of the enzyme in the presence of the inhibitor was compared to the absence of the inhibitor.

The I ₅₀ value was calculated from the plot of% inhibition versus inhibitor concentration.

Example 2

Synthesis of 3-acetylthiopropane oil-L-proline-t-butylester

0.865 g of 3-acetylthiopropanoic acid was dissolved in 2 ml of tetrahydrofuran (THF) which was re-distilled and cooled to 0 degrees Celsius. A cooling solution containing 1.2031 g of dicyclohexylcarbodiimide was added to 2 ml of THF, followed by addition of 1 g of cooled L-proline-t-butylester solution. The reaction mixture was stirred for 1 hour at 0 degrees Celsius and then overnight at 4 degrees Celsius.

The reaction mixture was filtered and the precipitate was washed with ethyl acetate and the solvent of the filtrate was removed under reduced pressure in a rotary evaporator.

The residue was dissolved in ethyl acetate and washed three times with cold 1N citric acid, twice with saturated NaCl solution, twice with cold 1N NaHCO ₃ and three times with saturated NaCl solution. The solution was dried over anhydrous MgSO ₄ and filtered.

This solvent was removed under reduced pressure in a rotary evaporator at 30 degrees Celsius to yield a colorless, transparent oily product which was approximately 87%. Thin layer chromatography was performed using five solvents and all of these products were shifted to one point.

Example 3

Synthesis of 3-mercaptopropane oil-L-proline

0.5 g of 3-acetylthiopropane oil-L-proline-t-butyl ester, a product obtained in Example 2, was mixed with 4.5 ml of 5.5N methanolic ammonia for 1 hour at room temperature and under nitrogen atmosphere to remove acetyl group. This solvent was then removed by rotary evaporator at 25 degrees Celsius.

The product was poured into methanol and redevaporated twice more on a rotary evaporator, then the clear oily residue was dissolved in ethyl ether, washed twice with 5% potassium bisulphate, once with saturated NaCl solution and then dried over MgSO ₄ . Filtered.

The residual solvent was removed in vacuo to yield a clear oily product that moved to one point as a result of thin layer chromatography using each of the three solvent systems.

The t-butylester protecting group was removed by reaction with trifluoroacetic acid in the anisol.

Example 4-6

2-acetylthiopropanoic acid, 3-acetyl thimeo-2-D-tilpropanoic acid, or 3-acetylthio-2-D, L-methylpropanoic acid instead of 3-acetylthiopropanoic acid of Example 2 The following compounds can be obtained by substituting for and performing the procedure of Examples 2 and 3 in substantially the same way. In the first step, dicyclohexylamine salts can be formed by removing the t-butylester protecting group using trifluoroacetic acid in the anisol, which helps to separate the isomers. Acetyl protecting groups can be raised in the second step using methanol ammonia as described in Example 3.

EXAMPLE

Example 7

Synthesis of 3-mercapto-2-methyl-propanoyl-L-3, 4-dehydrate proline

Dissolve 1 mmol of L-3, 4-dehydroproline (Δ ³ Pro) in DMF, cool the solution to minus 15 degrees Celsius, and neutralize this solution by adding 1 equivalent of N-ethylmorpholine.

In each reaction vessel at minus 10 degrees Celsius, a solution containing 1 equivalent of 3-acetylthio-2-methyl-propanoic acid in 1 equivalent volume of DMF was added in 1, 1'-carbonyldiimidazole. After mixing with the solution, the solution is stirred for 1 hour.

While maintaining the temperature at minus 10 degrees Celsius, the first solution containing △ ³ Pro and the second solution containing 3-acetylthio-2-methyl propanoic acid were mixed, and the mixed solution was then added at minus 10 degrees Celsius. Stir for time.

The solution is heated little by little to room temperature and then the solvent is removed using a rotary evaporator under reduced pressure at 40 degrees Celsius. Ethyl acetate (25 ml) is added and the solution is cooled to 0 degrees Celsius.

2 ml of 1N citric acid is added and the two phases are mixed and separated again. The phases, which are separated using a separatory funnel, are washed twice more with 2 ml of 1N citric acid and twice with saturated NaCl solution and finally dried over anhydrous MgSO ₄ .

This MgSO ₄ is removed by filtration and the solvent is removed using a rotary evaporator. The residue is dissolved and then recrystallized with a nonpolar solvent such as benzene to yield 3-acetylthio-2-D, L-methylpropane oil-L-3, 4-desuproline.

If 2-D-methyl isomer is required, the residue is dissolved in acetonitrile (approximately 3 ml) and the solution is warmed to 40 degrees Celsius.

One equivalent of dicyclohexylamine is added and the solution is left at room temperature overnight. The crystals can be collected by filtration and then recrystallized from isopropanol when a purer tablet is required for three washes with acetonitrile. The acetyl protecting group can be removed in the same manner as in Example 3.

Example 8-11

D, L-3, 4-dehydrate proline or L-3-hydroxyproline, or L-4-hydroxyproline or L-riazolidine-4-carboxylic acid instead of L-3, 4-dehydrate proline of Example 7. By substituting for and following the same procedure as in Example 7, the following compounds may be obtained.

EXAMPLE

Example 12

In a similar manner, 3-acetylthiopropanoic acid or 2-acetylthiopropanoic acid was substituted for the 3-acetylthio-2-methylpropanoic acid of Example 7-11. , 4-dehydrate proline, D, L-3, 4-dehydrate proline, L-3-hydroxyproline, L-4-hydroxyproline, and L-thiazolidine-4-carboxylic acid derivatives were obtained.

Example 13

^{N α - [3- (N α} - acetyl-phenyl -L- alanyl thio) -2-methylpropanoic five days D- Synthesis of -L- proline

A solution containing 41.5 mg of N ^α -acetyl-L-phenylalanine in 0.5 ml of re-distilled dimethylformamide (DMF) was cooled in an ice-dry ice-acetone bath at minus 20 degrees Celsius. To this solution was added a cold solution of 1.0 ml DMF containing 35 mg of 1, 1'-carbonyldiimidazole.

The solution was stirred for 2 hours at minus 10 degrees Celsius and then cold with 1 mg of DMF neutralized with N-ethylmorpholine containing 48 mg of 3-mercapto-2-D-methylpropane oil-L-proline. To the solution.

The reaction mixture was stirred at minus 10 degrees Celsius for another hour and then slowly heated to room temperature.

The solvent was removed under reduced pressure at 40 degrees Celsius and ethyl acetate was added to the residue.

The mixture was cooled in an ice bath, washed with 0.1N HCl and again washed three times with saturated NaCl solution.

This organic solvent was dried over anhydrous MgSO ₄ and then removed using a rotary evaporator.

This product was purified by Sephadex LH-20 ^tm column chromatography using a 1.2 cm x 95 cm column and eluted with isopropanol.

Only the top part was collected and the solvent was removed under reduced pressure to yield 35.5 mg of the title compound.

It was found by TLC using solvent systems 1, 2, 3, 5 that this product was clustered.

Example 14-21

Of Example 13 N ^{α-acetyl-a-phenylalanine} instead of N ^{α-acetyl-glycine,} N ^α-acetoxy butyl-alanine, N ^{α-acetyl-tryptophan,} N ^{α-acetyl-tyrosine,} N ^α-acetyl-a solution sour N ^α-acetyl-Lu Woosin, N ^α - acetyl-histidine or N ^α - acetyl - by carrying out the same procedure of example 13 is replaced with valine, and then to obtain the following compound.

EXAMPLE

Example 22

In a similar manner, the product of Examples 3-12 was substituted for the 3-mercapto-2-D-methylpropanoyl-L-proline of Examples 13-21 and the procedures of Example 13 were followed as is. 3,4-dehydrate proline, D, L-3, 4-dehydrate proline, L-3-hydroxyproline, L-4-hydroxyproline and L-riazolidine-4-carboxylic acid derivatives were obtained.

Example 23

L-phenylalanine, glycine, L-alanine, L-tryptophan, L-tyrosine, L-isosolecin, L-leucine, L-histidine and L-valine instead of the N ^α -acetyl derivatives of Examples 13-22 of N ^α - formyl, N ^α - propane five days, N ^α - butane five days, N ^α - phenylacetyl or N ^α - phenyl After a substituted propane five days induction chedeul and still perform the procedure of example 13, formyl, propane Oil, butan oil, phenylacetyl, and phenylpropane oil derivatives were obtained.

Example 24

^{N α - [3- (N α} - tert-butyloxycarbonyl -L- alanyl phenyl thio) -2-methylpropanoic five days D- Synthesis of -L- proline

A re-distilled DMF 0.5ml N ^α - tert-butyloxycarbonyl -L- phenylalanine ^{(N α -Boc-L-Phe} ) ice minus 20 ° C and the solution containing the 133mg degrees - on a dry ice-cooled in an acetone bath I was.

To this solution was added a cold solution of 1.0 ml of DMF containing 87 mg of 1,1'-carbonyldiimidazole, and then the solution was stirred for 2 hours at minus 10 degrees Celsius and then again N-ethyl mor To a cold solution containing 119.5 mg of 3-mercapto-2-2-D-methylpropane oil-L-proline in 1 ml of DMF neutralized with Pauline. The reaction mixture was neutralized with N-ethylmorpholine and stirred at minus 10 degrees Celsius for another hour and then slowly heated to room temperature.

The solvent was removed under reduced pressure at 40 degrees Celsius and ethyl acetate was added to this residue.

The mixture was cooled in an ice bath, washed with 0.1N HCl and three times with saturated NaCl solution.

This solvent was dried over anhydrous MgSO ₄ and then removed using a rotary evaporator. The product was purified by liquid chromatography with Sephadex G-10 ^tm using a 1.2 cm x 95 cm separator tube and eluted with a solution of THF: isopropane oil 3: 7 (volume ratio).

Only the top portion was collected and the solvent was removed under reduced pressure to yield 165 mg of the designated product.

This product was found to be homogeneous by paper electrophoresis at pH 5.0 and TCL using solvent systems 1, 2 and 3.

Example 25-32

Example 24 of the N ^α -Boc-Phe with N ^α -Boc- glycine, N ^α -Boc- alanine, N ^α -Boc- tryptophan, N ^α -Boc- tyrosine, N ^α -Boc- this new solution, N ^α -Boc- base Woosin, N ^α -Boc- Hi Stephen Dean or resulting from replacing the N ^α -Boc-valine and as to perform the procedure of example 24 to give the following compounds.

EXAMPLE

Example 33

In a similar manner, the 3-mercapto-2-D-methylpropaneoyl-L-proline of Example 24-32 was replaced with the product of Example 3-12 and the procedure of Example 24 was carried out as is L-3. , 4-dehydrate proline, D, L-3, 4-dehydrate proline, L-3-hydroxyproline, L-4-hydroxyproline, and L-thiazolidine-4-carboxylic acid derivatives were obtained.

Example 34

Synthesis of N ^α -cyclopentylcarbonyl-L-phenylalanine

A cold solution containing 2.06 gm of dicyclohexylcarbodiimide in 10 ml of dichloromethane was added to a solution containing 1.414 gm of cyclopentanecarboxylic acid in 5 ml of dichloromethane at minus 5 degrees Celsius.

Then a solution containing 4.28 gm of L-phenylalanyl benzoyl ester toluene sulfone salt in 10 ml of DMF neutralized with 1.36 ml of -ethyl morpholine was added.

The reaction mixture was stirred for 1 hour at 0 degrees Celsius and then left for 3 hours at room temperature.

Dicyclohexyl urea was removed by filtration and 50 ml of ethyl acetate was added to the filtrate.

The organic phase was washed until neutral and dried over anhydrous MgSO ₄ and filtered. The solvent was removed using a rotary evaporator and the residue was crystallized from isopropanol and hexane to yield 2.35 gm of white crystals with a melting point of 88-89 degrees Celsius.

The results of elemental analysis of these crystals are as follows.

Calculated: C = 75.19; H = 7.17; N = 3.9855

Found: C = 74.96; H = 7.17; N = 4.09

The sterilizer was removed from benzyl by hydrolyzing the carbon phase of anhydrous alcohol with 2 gm of 10% palladium.

This catalyst was removed by filtration and ethanol was removed using a rotary evaporator.

The residue was crystallized with ether and hexane to yield 1.15 gm of the designated product of white color with a melting point of 107-108 degrees Celsius.

Elemental analysis of this crystal gave the following results.

Calculated: C = 68.94; H = 7.33; N = 5.36

Found: C = 68.90; H = 7.32; N = 5.34

The product was found to be homogeneous by paper electrophoresis at pH 1.9 and pH 5.0 or by TLC using solvent systems 1, 2 and 3.

This designated product can be weakly represented by N ^α -cpc-L-Phe.

Example 35

^{N α - [3- (N α} - cyclopentyl-carbonyl-phenyl -L- alanyl thio) -2-methylpropanoic five days D- Synthesis of -L- proline

A solution containing 52.5 mg of the product obtained in Example 34 in 0.5 ml of re-distilled DMF was cooled in an ice-dry ice-acetone bath at -20 degrees Celsius. To this solution was added a cold solution containing 34 mg of 1,1'-carbonyldiimidazole in 1.0 ml of DMF.

The solution was stirred at -10 degrees Celsius for 2 hours and then cold solution containing 45.6 mg of 3-mercapto-2-D-methylpropane oil-L-proline in 1 ml of DMF neutralized with N-ethyl morpholine. Mixed. The reaction mixture was stirred at -10 degrees Celsius for another hour and heated slowly until room temperature.

The solvent was removed under reduced pressure of 40 degrees Celsius and ethyl acetate was added to this residue, then the mixture was cooled, washed with 0.1N HCl and again washed with saturated NaCl solution.

The solvent was dried over anhydrous MgSO ₄ , removed using a rotary evaporator, and the product was purified by Sephadex LH-20 ^tm tube chromatography using a 1.2 cm × 95 cm separation tube and eluted with isopropanol.

The top portion was combined and the solvent removed under reduced pressure to yield 60.5 mg of the title compound. TLC using solvent systems 1, 2, 3 and 5 revealed that the product was homogeneous.

Example 36-43

Substitution with the benzoyl ester toluenesulfonate of glycine, L-alanine, L-tryptophan, L-tyrosine, L-isoleucine, L-leucine, L-histidine or L-valine instead of the L-phenylalanine salt of Example 34 The following compounds were obtained by performing the following procedures of Examples 34 and 35 as they were.

EXAMPLE

Example 44

In a similar manner, the 3-mercapto-2-D-methylpropaneoyl-L-proline of Examples 35-43 was replaced with the product of Examples 3-12 and the procedure of Example 35 was followed as is L-3, 4-dehydrate proline, D, L-3, 4-dehydrate proline, L-3-hydroxyproline, L-4-hydroxyproline, and L-thiazolidine-4-carboxylic acid derivatives were obtained.

[45]

Synthesis of N ^α- (3-L-phenylalanylthio-2-D-methylpropaneoyl) -L-proline

The product obtained in Example 24 can be removed by stirring a mixture of 30 mg of this product, 50 μl of anisol and 200 μl of anhydrous trifluoroacetic acid (TFA) at room temperature for 1 hour. Anisole and TFA were removed under reduced pressure of 35 degrees Celsius and the residue was triturated with anhydrous ether.

This white residue was purified by liquid chromatography with Sephadex G-10 ^tm using a 1.2 cm x 95 cm separation tube, then eluted with 5% acetic acid. Only the top portion was collected and lyophilized to yield 17.5 mg of the designated compound.

The product was homogeneous, either by electrophoresis at pH 1.9 and 5.0 or by TLC using solvents 4 and 5.

Example 46-53

In a similar manner, the procedure of Example 45 was followed to remove the protecting groups of the product obtained in Examples 25-32 to afford the following compounds.

EXAMPLE

Example 54

In a similar manner, L-3,4-dehydrate proline, D, L-3, 4-dehydrate proline, L-3-hydroxyproline, L-4-hydroxyproline by removing the protecting group of the compound described in Example 33. And L-thiazolidine-4-carboxylic acid derivatives.

The protecting group removal is carried out by following the procedure of Example 45.

Example 55

Synthesis of N-hydroxysuccinimide Ester of Cyclopentane Carboxylic Acid

A cold solution containing 11.4 gm of dicyclohexylcarbodiimide in DMF was added dropwise to the mixture containing 5.76 gm of N-hydroxysuccinimide and 5.71 gm of cyclopentanecarboxylic acid in DMF at 0 degrees Celsius.

The reaction mixture was stirred for 30 minutes at 0 degrees Celsius and then left overnight at 4 degrees Celsius.

The crystalline dicyclohexyl urea was removed by filtration and the precipitate was washed with ethyl acetate.

The solvent in the mixed filtrate was removed under reduced pressure and the residue was crystallized with benzene and hexane to yield 5.77 gm of white crystals having a melting point of 72.5-73 degrees Celsius.

The infrared absorption spectrum in chloroform showed the typical spectrum of N-hydroxysuccinimide ester.

Elemental analysis of these crystals yielded the following results.

Calculated: C = 56.86; H = 6.20; N = 6.63

Found: C = 56.77; H = 6.07; N = 6.66

Example 56

Synthesis of N ^α -cyclopentylcarbonyl-N ^ε -tert-butyloxycarbonyl-L-lysine

10ml of water and NaHCO ₃ 420mg wave in THF 5ml N ^ε - tert - the solution containing the butyloxycarbonyl -L- lysine 1.2316gm was cooled with stirring in an ice bath.

To this solution was added a cold solution containing 1.199 m of the product obtained in Example 55 in 10 ml of THF.

After the reaction mixture was stirred at room temperature overnight, THF was removed using a rotary evaporator at 35 degrees Celsius.

About 20 ml of water was added to this reaction mixture and adjusted to a pH of 9 with solid NaHCO ₃ .

The aqueous phase was extracted three times with ethyl acetate and the organic phase was discarded. This aqueous solution was cooled in an ice bath and acidified to pH 2 with 1N HCl in the presence of ethyl acetate.

The organic phase was washed twice with ice water and again with saturated NaCl solution. The organic solution was dried over anhydrous MgSO ₄ and filtered. The solvent was removed with a rotary evaporator and the residue was crystallized with ether and hexane to yield 1.135 gm of white crystals with a melting point of 104.5-105.5 degrees Celsius.

The results of elemental analysis of these crystals are as follows.

Calculated: C = 59.63; H = 8.83; N = 8.18

Found: C = 59.74; H = 8.85; N = 8.24

This product was known to be homogeneous by paper electrophoresis at pH 1.9 and pH 5.0 or by TLC using solvent systems 1, 2 and 3.

Example 57

Synthesis of N ^α -cyclopentylcarbonyl-N ^ε -tert-butyloxycarbonyl-L-lysine-N-hydroxysuccinimide ester

A solution containing 346 mg of N-hydroxysuccinimide and 1.027 gm of the product obtained in Example 56 in 10 ml of CH ₂ Cl ₂ was cooled to -50 degrees Celsius.

The reaction mixture was stirred for 30 minutes at 0 degrees Celsius and left overnight at 4 degrees Celsius. The crystalline dicyclohexyl urea was removed by filtration and washed with ethyl acetate.

This mixed filtrate was washed twice with 1N NaHCO ₃ , twice with water and finally with saturated NaCl solution.

The organic phase was dried over anhydrous MgSO ₄ , filtered and the solvent removed using a rotary evaporator.

The residue was crystallized with isopropanol to yield 0.844 gm of white crystals having a melting point of 140.5 degrees Celsius.

Infrared absorption spectra in chloroform showed typical spectra of N-hydroxysuccinimide esters.

Elemental analysis of these crystals gave the following results.

Calculated: C = 57.39; H = 7.57; N = 9.56

Found: C = 57.10; H = 7.57; N = 9.61

Example 58

Synthesis of N ^α -cyclopentylcarbonyl-N ^ε -tert-butyloxycarbonyl-L-lysyl-L-phenylalanine

A solution containing 220 mg of the product obtained in Example 57 in 2 ml of dioxane was added dropwise to a mixture containing 51 mg of NaHCO ₃ and 99.1 mg of L-phenylalanine in a mixture of 2 ml of water and 1 ml of DMF.

The reaction mixture was stirred overnight at room temperature and dioxane was removed using a rotary evaporator at 35 degrees Celsius.

Ethyl acetate (10 ml) was added to this reaction mixture, cooled, acidified to pH 2 with 0.1 N HCl and then the aqueous solution was discarded.

The organic phase was washed with ice water and saturated NaCl solution, dried over anhydrous MgSO ₄ and the solvent removed by rotary evaporator.

The residue was crystallized with ether and petroleum ether (b.p. 30-60 ° C.) to yield 95 mg of a white solid with a melting point of 90-92 degrees Celsius.

This product was known to be homogeneous by paper electrophoresis at pH 1.9 and pH 5.0 and TLC using solvent systems 1, 2, 3 and 4.

The result of elemental analysis is as follows.

Calculated: C = 63.78; H = 8.03; N = 8.58

Found: C = 63.40; H = 8.07; N = 8.34

Example 59

Of N ^α- [3- (N ^α -cyclopentylcarbonyl-N ^δ -tert-butyloxycarbonyl-L-lysyl-L-phenylalanylthio) -2-D-methylpropaneoyl] -L-proline synthesis

A solution containing 73.5 mg of the product obtained in Example 58 in 0.5 ml of re-distilled DMF was cooled in an ice-dry ice-acetone bath at -20 degrees Celsius.

To this solution was added a cold solution containing 26 mg of 1,1'-carbonyldiimidazole in 1.0 ml of DMF.

The solution was stirred for 2 hours at -10 degrees Celsius and then mixed with a cold solution containing 36 mg of 3-mercapto-2-D-methylpropane oil-L-proline in 1 ml of DMF neutralized with N-ethylmorpholine. It was.

The reaction mixture was stirred for an additional hour at -10 degrees Celsius and then heated slowly until room temperature. The solvent was removed under reduced pressure at 40 degrees Celsius and ethyl acetate was added to this residue.

The mixture was cooled in an ice water bath, washed with 1N citric acid and washed three times with polyvinyl saturated NaCl solution.

The solvent was dried over anhydrous MgSO ₄ and then removed using a rotary evaporator. This product was purified by Sephadex LH-20 ^tm tube chromatography using a 1.2 cm × 95 cm separator tube and eluted with THF: isopropanol 3: 7 (volume ratio).

Top section only collected together and the solvent is removed under reduced pressure ^{N α - [3- (N α} -cpc-N δ -Boc-L-Lys-L-Phe- thio) -2-methylpropane-D- five days; - L-proline yielded the designated product which could be weak.

Example 60

^{N α - [3- (N α} - cyclopentyl carbonyl -L- florisil -L- alanyl phenyl thio) -2-methylpropanoic five days D- Synthesis of -L- proline

N-Boc groups were removed from lysine by stirring a mixture of 30 mg of the product obtained in Example 59, 50 μl of anisole, and 200 μl of anhydrous trifluoroacetic acid (TEA) at room temperature for 1 hour.

Anisole and TFA were removed under reduced pressure of 35 degrees Celsius and the residue was triturated with anhydrous ether.

The residue was purified by liquid chromatography with Sephadex G-10 ^tm using a 1.2 cm × 95 cm separator and eluted with 5% acetic acid.

Only the top part is collected and lyophilized to give N ^α- [3- (N ^α- [3- (N ^α -cpc-L-Lys-L-Phe-thio) -2-D-methylpropane oil] -L-proline It yielded the designated product which could be weak.

Example 61-68

The L-phenylalanine of Example 58 was substituted with Glycine-L Alanine, L-Tryptophan, L-Tyrosine, L-Isoleucine, L-Lucine, L-Histidine or L-Valine and the compounds of Examples 58, 59, 60 As a result of the procedure, the following compounds were obtained.

EXAMPLE

Example 69

In a similar manner, in Example 59 and 61-68 3-mercapto-2-D-methylpropaneoyl-L-proline was replaced with the product of Examples 3-12 and the procedures of Examples 59 and 60 were carried out as is. L-3, 4-dehydrate proline, D, L-3, 4-dehydrate proline, L-3-hydroxyproline, L-4-hydroxyproline and L-thiazolidine-4-carboxylic acid derivatives were obtained.

Example 70

The cyclopentanecarboxylic acid of Example 55 was substituted with pyro-L-glutamic acid and the procedures of Examples 55-57 were different as is. N ^α -pyro-L-glutamyl-N ^δ -tert-butyloxycarbonyl-L-lysine -N-hydroxysuccinimide ester was obtained.

The fatigue-L-glutamyl derivative as a result of replacing the N ^α -cpc- N ^δ -Boc-L-Lys-N-hydroxysuccinimide ester of Example 58 with this product and following Examples 58-69, Got it.

Example 71

Process for preparing pyro-L-glutamyl-L-phenylalanine benzyl ester

A solution containing 0.52 g of pyro-L-glutamic acid, 1.72 g of L-phenylalanine benzyl ester toluenesulfonic acid and 0.55 ml of N-ethylmorpholine in 5 ml of DMF and 20 ml of dichloromethane was cooled in an ice bath with stirring.

A solution containing 0.826 g of dicyclohexylcarbodiimide in 2 ml of dichloromethane was added to the reaction mixture above. The reaction mixture is stirred in iced water for 1 hour and left at room temperature overnight.

Dicyclohexyl urea was removed by filtration and the product was washed with ethyl acetate.

The solvent in the mixed filtrate was removed under reduced pressure using a rotary evaporator at 40 degrees Celsius, ethyl acetate (25 ml) was added to this residue and the organic solution was washed until neutral. This organic phase was dried over anhydrous MgSO ₄ , filtered and the solvent removed using a rotary evaporator.

The material was crystallized with isopropanol and ether to yield 1.01 g of white needles with a melting point of 103-104.5 degrees Celsius.

This material was homogeneous in both TLC and electrophoretic systems.

The result of elemental analysis is as follows.

Calculated: C = 68.84; H = 6.05; N = 7.64

Found: C = 68.58; H = 6.05; N = 7.56

Example 72

Process for preparing pyro-L-glutamyl-L-phenylalanine

The benzoyl ester protecting group of the compound of Example 71 (1.0 g) was removed by catalytic hydrogenolysis of 15 ml of ethanol and 0.15 ml of glacial acetic acid overnight at room temperature of H ₂ 20 psi using 150 mg of 10% (by weight) Pd. It was.

This catalyst was removed by filtration and the solvent removed by rotary evaporator. The material was crystallized with isopropanol and benzene to yield a total of 402 mg of white crystals with a melting point of 147-149 degrees Celsius.

Electrophoresis at pH 1.9 and pH 5.0 and TLC using solvent systems 1, 2, and 3 resulted in a homogeneous material.

The result of elemental analysis is as follows.

Calculated: C = 60.86; H = 5.84; N = 10.14

Found: C = 60.37; H = 5.85; N = 9.98

Example 73

Method for preparing N ^α- [3- (Pyro-L-glutamyl-L-phenylalanylthio) -2-D-methylpropane oil-L-proline

A solution containing 87 mg of 1,1'-carbonyldiimidazole in 1.0 ml of DMF was added to a solution containing 139 mg of the product of Example 72 in 0.5 ml DMF at -15 degrees Celsius.

The reaction mixture was stirred for 1 hour at -10 degrees Celsius and a mixed solution containing 0.072 ml of N-ethyl morpholine and 119.5 mg of 3-mercapto-2-D-methylpropane oil-L-proline in 1 ml of DMF. Was added. The reaction mixture was stirred for an additional hour at -10 degrees Celsius and then slowly heated to room temperature.

DMF was removed under reduced pressure with a rotary evaporator at 40 degrees Celsius and 7 ml of ethyl acetate and 2 ml of 1N citric acid were added.

The organic phase was washed three times with 1N citric acid and twice with saturated NaCl solution. The organic phase was dried over anhydrous MgSO ₄ and filtered.

The solvent was removed using a rotary evaporator and the residue was purified by Sephadex G-25 ^tm (1.2 × 99 cm) partition tube chromatography with n-butanol: acetic acid: H ₂ O (volume ratio 4: 1: 5).

This product was homogeneous as a result of electrophoresis at pH 5.0 with TLC (solvents 1, 2, 3).

Example 74

Pyro-L-glutamic acid may be referred to as N ^α , -N ^δ -bis-t-buryloxycarbonyl-L-lysine (hereinafter referred to as bis-Boc-L-Lys) or N ^α , N ^γ , N ^ω -triacetinyl By replacing with oxycarbonyl-L-arginine (hereinafter referred to as tri-Adoc-L-arginine) and following the procedure of Example 71, bis-Boc-L-Lys of the corresponding L-phenylalanine-benzyl ester or tri-Adoc-L-Arg derivatives can be synthesized.

Bis-Boc-L-Lys is commercially available and Tri-Adoc-L-Arg is described by Jager, G. and Geiger, R., Chem. Ber. 103, 1727 (1970).

Example 75

In the procedures of Examples 71 and 74 by replacing benzyl esters of glycine, alanine, tryptophan, tyrosine, isoleucine, leucine, histidine or valine with the corresponding fatigue-L-glutamyl, bis-Boc-L-Lys and tri-Adoc-L-Arg derivatives were obtained.

The benzyl ester protecting group was essentially removed as described in Example 72.

Example 76

Bis-Boc-L-Lys and tri-Adoc-L- as a result of reacting the compounds produced in Examples 72, 74, and 75 with some of the compounds obtained in the procedure of Example 3-12 and following the procedure of Example 73 as they are Arg derivatives were calculated.

The bis-Boc and tri-Adoc protecting groups were removed by treatment with trifluoroacetic acid in anisol.

By performing the methods and procedures of Examples 2-12 and 71-76 as they were, compounds of the thioester group having the following general formulas were obtained.

In the food,

R is L-arginyl, L-lysyl or pyro-L-glutamyl;

A is L-phenylalanyl, glycal, L-alanyl, L-tryptiphyl, L-tyrosyl, L-isosolyl, L-leusil, L-histidyl, or valely and their α-amino groups Is an amide bond with R;

R ₁ is hydrogen or methyl;

와 이미드 결합하고 있다;R ₂ is L-proline, L-3, 4-dehydrate proline, D, L-3, 4-dehydrate proline, L-3-hydroxyproline, L-4-hydroxyproline or L-thiazolidine-4 -Carboxylic acids and their imino groups

And imide bond;

n is 0 or 1 and when n = 0 R ₁ is methyl.

Example 77

^{N α - [3- (N α} - benzoyl -L- tryptophyl thio) -2-methylpropanoic five days D- Synthesis of -L- proline

A solution containing 62 mg of N-benzoyl-L-tryptophan in 0.5 ml of re-distilled dimethylformamide (DMF) was cooled in an ice-dry ice acetone bath with -20 degrees brow seed.

To this solution was added a cold solution containing 35 mg of 1, 1 'carbonyldiimidazole in 1.0 ml of DMF and then stirred for 2 hours at -10 degrees Celsius, and 1 ml of DMF neutralized with N-ethylmorpholine. To a cold solution containing 48 mg of 3-mercapto-2-D-methylpropane oil-L-proline in water.

The reaction mixture was stirred for an additional hour at -10 degrees Celsius and then heated slowly until room temperature.

This solvent was removed under reduced pressure at 40 degrees Celsius and ethyl acetate was added to this residue.

The mixture was cooled in an ice bath, washed with 0.1N HCl and then washed three times with saturated NaCl solution.

This solvent was dried over anhydrous MgSO ₄ and then removed by rotary evaporator.

The product was purified by liquid chromatography with Sephadex × LH-20 ^tm using a 1.2 cm × 95 cm separator tube and eluted with isopropanol. Only the top part was collected and the solvent was removed under reduced pressure, yielding 60.5 mg of a fizzing compound, a foamy material.

This product was found to be homogeneous by TLC using solvent systems 1, 2, 3 and 5.

Example 78-85

The N ^α -benzoyl-L-tryptophan of Example 77 was treated with N ^α -benzoyl-L-tyrosine, N ^α -benzoyl-L-histidine, N ^α -benzoyl-phenylalanine, N ^α -benzoyl-glycine, benzoyl-L-alanine , N ^α - benzoyl -L- this new solution, N ^α - benzoyl -L- Lou during or N ^α - by a substituted benzoyl -L- valine and as to perform the process of example 77 to give the following compounds.

EXAMPLE

Example 86

In a similar manner, L-3 by replacing the 3-mercapto-2-D-methylpropaneoyl-L-proline of Examples 77-85 with the products of Examples 3-12 and following the procedures of Example 77 as is , 4-dehydrate proline, D, L-3, 4-dehydrate proline, L-3-hydroxyproline, L-4-hydroxyproline, and L-thiazolidine-4-carboxylic acid derivatives were obtained.

^{N α - [3- (N α} - benzoyl-phenyl alanyl thio) Another process for preparing a D- -2-methylpropane-five days -L- proline is described in the following two examples.

Example 87

Method for preparing N ^α -benzoyl-phenylalanine-N-hydroxysuccinimide ester

1.347 g of benzoyl-phenylalanine and 0.576 g of N-hydroxysuccinimide were mixed in a THF and DMF 1: 1 (volume ratio) mixed solution.

This mixed solution was incubated overnight in the presence of 1.133 g of dicyclohexylcarbodiimide at 4 degrees Celsius.

The reaction mixture was filtered and solvent removed under reduced pressure at 30 degrees Celsius.

The remaining white residue was recrystallized with THF-isopropyl alcohol to yield 1.194 g (65.2 g) of a white solid having a melting point of 156-157 degrees Celsius.

Infrared absorption spectra in the chloroform is 1818cm ^-1 3440cm ^-1-carboxy succinimide and N- represents an NH represents the characteristics of the imide group, 1790cm ^-1, 1744cm ^-1 and 1669cm ^-1 in the characterizing part of N- benzoyl Showed the girdle.

Example 88

^{N α - [3- (N α} - benzoyl-phenyl alanyl thio) Synthesis of D- -2-methylpropane-five days -L- proline

93.3 mg of 2-D-methyl-3-mercaptopropaneoyl-L-proline prepared by the method described in Example 87, 62 mg of 1-hydroxybenzotriazole (HOBt), N-benzoyl-phenylalanyl-N- The reaction mixture containing 165 mg of hydroxysuccinimide ester was cooled in a large 0 ° C. ice bath and then 0.0544 ml of N-ethylmorpholine was added. The reaction mixture was stirred for 3 hours in an ice bath, incubated overnight at 4 degrees Celsius, and then left at room temperature for 20 hours.

The reaction was terminated by addition of 25 μl of N, N-dimethyl-1, 3-propanediamine and stirred for an additional 2 hours.

Ethyl acetate was added to the reaction mixture, extracted with cold 0.1N HCl, washed, twice with water and three times with saturated NaCl solution. The mixture was dried over anhydrous MgSO ₄ and filtered.

The solvent was removed under reduced pressure in the rotary evaporator to yield a clear oily product.

This product was purified by chromatography with a Sephadex LH-20 ^tm separator tube 1.2 cm × 96 cm, which was eluted with THF to collect 2.5 ml of fragments.

Sections 30 and 31 were put together and the solvent was removed under high vacuum to yield 110 mg of product.

Anhydrous ether was added to obtain a white substance such as gum.

The ether was poured still and a white substance such as gum was dissolved in THF, transferred to a glass jar, dried in a stream of nitrogen and dried overnight with P ₂ O ₅ .

As a result, 82 mg of foamy product was obtained.

Sections 29, 32 and 33 were chromatographed again under the same conditions and second chromatographed fragments 33-36 were carried out as described previously to yield 51 mg of product. The yield was 71%.

N ^α - is described in [propane five days 2- (N ^α - benzoyl-phenyl alanyl thio)] -L-3, 4- dehydration proline with another method for producing the derivatives of the following two examples.

Example 89

Synthesis of 2-benzoylphenylalanylthiopropanoic acid

The solution containing benzoyl-phenylalanine in 30 ml of re-distilled dimethylformamide (DMF) was cooled to -20 degrees Celsius.

In 10 ml of re-distilled DMF, a solution containing 1,1'-carbonyldiimidazole was added dropwise while shaking vigorously. The temperature should not exceed -14 degrees Celsius.

After addition the solution was stirred at -10 degrees Celsius for 2 hours.

D and L-thiolactic acid contained in re-distilled DMF neutralized with re-distilled N-ethylmorpholine was added with continuous stirring at 10 ° C for 1 hour. The solution was slowly heated to room temperature and then approximately the same volume of ethyl acetate was added. The mixture was cooled and neutralized with concentrated HCl in saturated NaCl.

The organic phase was washed three times with a less saturated NaCl solution, ie, a solution diluted with saturated NaCl: water to 5: 1 (volume ratio).

In some cases, a three-layer system is found, in which case the intermediate layers are collected and mixed with the aqueous phase of the lower layer. The organic phase was dried over anhydrous MgSO ₄ , filtered and placed in a rotary evaporator to remove the solvent.

The combined aqueous and intermediate phases were acidified to pH 2 with concentrated HCl at 0 ° C. and extracted three times with ethyl acetate.

The organic phase was washed with saturated NaCl, dried over anhydrous magnesium sulfate, filtered and evaporated on a rotary evaporator to recover clear oil.

Example 90

^{N α - [2- (N α} - benzoyl phenyl alanyl-thio-propane-five days)] - L-3, the synthesis of 4-muddy water-proline

Boc-L-3 and 4-dehydroproline (213 mg; 1 mmol) were reacted with 1 ml of anhydrous trifluoroacetic acid for 1 hour to remove the protecting group.

Anhydrous 3 ml of ethyl ether was added and the solvent was removed using a rotary evaporator at 30 degrees Celsius.

The residue was washed three times with ether and dried over NaOH in a vacuum drier.

The solution containing 358 mg (1 mmol) of the product obtained in Example 89 in 3 ml of re-distilled DMF was cooled to -20 degrees Celsius in a dry ice-acetone bath. A solution containing 171 mg (1.05 mmole) of 1,1'-carbonyldiimidazole was added dropwise to 1 ml of re-distilled DMF and the temperature of the reaction mixture was maintained at -10 to -15 degrees Celsius.

The reaction mixture was stirred for 2 hours and then a cold solution containing L-3, 4-dehydroproline trifluoro acetate in 2 ml of DME (neutralized with triethylamine) was added.

The reaction mixture was heated slowly to room temperature.

The solvent was removed by rotary evaporator at high vacuum at 35 degrees Celsius and the residue was dissolved in 10 ml of ethyl acetate and ₂ ml of H ₂ O. The mixture was acidified with 1N HCl at 0 ° C. to pH 2 and mixed.

The organic phase was separated and collected and the aqueous phase was extracted three more times with 5 ml of ethyl acetate.

The combined organic phases were decolorized with heart char washed twice with H ₂ O and three times with saturated NaCl.

This organic phase was dried over anhydrous MgSO ₄ and filtered.

This filtrate was evaporated to dryness in a rotary evaporator to obtain 366 mg of an oily product.

This product was purified by chromatography on equilibrated Sephadex LH-20 ^tm (1.2 × 98 cm) and eluted with methanol.

150 drops of fragments (2.9 ml) were collected to elute the desired compound in fragments 24-29.

The solvent was removed by rotary evaporation and further purified by chromatography once more with Sephadex LH-20 ^tm .

The second separator was equilibrated and eluted with isopropanol to collect only the top part and the solvent was removed by rotary evaporator.

This product had approximately 95% purity as measured by thin layer chromatography using solvent systems (2) and (3).

Example 91

The inhibitory effect of the various compounds synthesized in vitro was measured in the analytical system described in Example 1.

The prepared enzyme was Ryan. Purified from human urine as described by J.W., et al., Tissue and Cell 10, 555 (1978).

Table 1 shows the concentrations of various compounds of I ₅₀ indicates whether inhibitor required for 50% inhibition of the enzyme at standard quantitative gyeha, which contains a lower level of the substrate ₅₀ than I go.

TABLE 1

Example 92

Oral Dosing Effect of Various Compounds

Mice (210-290 g body weight) were fasted overnight and then anesthetized with intraperitoneal pentobarbital (50-60 mg / kg).

After the tracheotomy, the animals mechanically shot clear air. A tube was inserted into the vein of the femoral to inject angiotensin I and another was inserted into the common carotid artery to directly measure arterial blood pressure.

1000 units of heparin were injected through the femoral vein to prevent humoral coagulation.

Blood pressure was measured with a blood pressure transducer connected to a polygraph.

Rats were injected with a solution containing 160-400 ng / kg of angiotensin I in 20 μl of 0.9 g% NaCl. The amount of angiotensin I was sufficient to increase the mean arterial blood pressure by 27-50 mmHg.

After the injected mice responded to anginotensin I, 5.5-23 μmole / kg of each compound (the drug was dissolved in 10 μl of H ₂ O 0.15 ml + NaHCO ₃ ) was injected via gavage.

At regular intervals, the effects of angiotensin I, 160-400 ng / kg, on mean arterial blood pressure were tested and the results are shown in Table 2. Doses of specific compounds are listed in parentheses below the compound number.

For example, 13 (13.6) means that 13.6 μmole / kg of the compound obtained in Example 13 was taken orally.

When testing the effects of angiotensin I, the time after oral administration of a particular compound is expressed in minutes, which is written in [] after the number in% of inhibition.

TABLE 2

Example 93

Intravenous effects of various compounds

Anesthetized mice were prepared as described in Example 92. After rats responded to angiotensin I, each compound containing 2 μmole / kg in a volume of 15 μl of C.OIN sodium bicarbonate was injected through the femoral vein.

At regular intervals, the effect of angiotensin I, 400 ng / kg on mean arterial blood pressure was tested. The results are shown in Table 3, and Table 3 follows the model shown in Table 2.

TABLE 3

Although the present invention has been described in connection with specific aspects thereof, many modifications to the present invention are possible so that this application uses the arguments obtained in this paper, for example, conventional methods well known in the art to which the present invention pertains. It will be appreciated that it is intended to speak of a variant use or application of the invention in accordance with the principles of the invention in general, including that it may be applied to the essential features described previously and in accordance with the limitations of the appended claims.

Claims (1)

The compound of formula (II) is reacted with R ₂ -H to form a compound of formula (III), and R ₃ is removed from the compound of formula (III) to form a compound of formula (IV). A process for producing S- (acylamidoacyl) mercaptoacyl proline of formula (I), wherein the compound of formula (IV) is reacted with RA-OH.

Among these expressions. R is hydrogen, formyl, acetyl, propane oil, butane oil, phenylacetyl, phenyl propane oil, benzoyl, cyclopentylcarbonyl, tert-butyloxycarbonyl, cyclopentylcarbonyl-L-lysyl, pyro-L-glu Tamil-L-lysyl, L-arginyl, L-lysyl or pyro-L-glutamyl, A, L-phenylalanyl, glycyl, L-alanyl, L-tryptiphyl, L-tyrosyl, L-isorucil, L-leusil, L-histidyl or L-valyl, The amino group is an amide bond with R. When R is benzoyl, phenylalanyl must be racemic, R ₁ is hydrogen or methyl, R ₂ is

And these amino groups are adjacent

And amide bonds, R ₃ is acetyl, n is 0 or 1 and when n = 0 R ₁ is methyl.