PL150055B1 - A method of obtaining new acylpeptides - Google Patents

Description

PATENT DESCRIPTION

150 055

POLAND

REPUBLIC

FOLK

Additional patent to patent no. Applied for: 86 11 24 / P. 262563 /

11 25 for claims 1-3 Priority _{86 08 27} for claims 4-6

United States of America

IS A HEAT

Born / Patent

Int. Cl. ⁴ C07K 5/04

OFFICE

PATENT

PRL

Application announced: 04 28 Patent description published: 90 07 31

Inventor

Patent holder: Pfizer Inc, New York / United States of America /

METHOD OF MAKING NEW ACYLOPEPTIDES

The subject of the invention is a method for the production of new acylpeptides containing glutamic acid useful as immunostimulants and anti-infective agents in the form of pharmaceutical compositions for the treatment of infections. This relatively new field of immunopharmacology, and especially the part of it dealing with immunomodulation, is rapidly developing further.

A number of naturally occurring compounds have been studied, including the tetrapeptide - tuftsin, known as the chemical compound: N ² - / 1- / N2-L-threonyl-L-lysyl / -L-prolyl / -L-arginine

Much attention has been given to synthetic peptidoglycan derivatives, especially those known as muramyl dipeptides.

A comprehensive discussion of the numerous compounds tested as immunomodulators, and especially as immunostyrucants, can be found in the publications: Duker and ln · Annu · Rep · Med · Chem · 14, 146-167 / 1979 /: Lederev, 0. Med · Chem. 23, 819-825 / 1980 / and O. Kralovec, Drugs of the Futura 8, 615-638 / 1983 /.

Peptide immunostimulants have been described in numerous patents:

-N-acyl alpha-glutaroyl-L-alanyl dipeptides in German Patent Specification No. 3,024,355, published January 15, 1981;

- tatra- and pentapeptides containing O-alanyl-L-glutamyl units and L-elanyl-D-glutamyl units, respectively, in British patent specification No. 2,053,231, published February 4, 1981 and in German patent specification No. 3,024,261, published January 8, 1981

- derivatives of N-acyl-alanyl-gema-D-glutemyl tripeptides, in which the C-terminal amino acid is lysine or diaminopimelic acid in German Patent Specification No.

024 369, published January 15, 1981, and

- lactol-tatrapeptides consisting of N-lactylolalanyl, glutaminopimelin and carboxymethylamino members in EP-11 283,

150 055

150,055, published May 23, 1980. · Further polypeptide immunostimulants of general formula 4 in which R ¹ is hydrogen or acyl, R ² is, inter alia, hydrogen, lower alkyl, hydroxyethyl, benzyl; R ³ and R ^{4 are} each hydrogen, carboxyl, -COR ⁷ R® where R ⁷ is hydrogen, lower alkyl, optionally substituted with hydroxy, and R® is monodicarbokylower alkyl, R ⁵ is hydrogen or carboxyl provided that if one of R ⁴ or R ⁵ is hydrogen, the other is carboxyl or -C0R ⁷ R®, R ⁶ is hydrogen, m is 1-3, n is 0 to 2 1 their derivatives in which the carboxyl and amino groups are protected are described in US patents 4 311 640, 4 322 341 and in European patent applications 50856, 51812, 53388, 55846 1 57419.

None of the polypeptides disclosed in the prior art contain a heterocyclyl residue at a position substituted by a different R in the formula above.

Kitaura et al. In □ Med Chem 25, 335-337 / 1962 / report about N ² - / gema-Dglutamyl / -meso-2 / L /, 2 / O / -diaminopimelinic acid as having a minimal structure capable of eliciting a biological response characteristic of compounds of formula 4, wherein η · 1, R ¹ is CH ₃ CH / OH / -C0-; R ² is CH 3, R ³ and R ⁵ is -COOH; R ⁴ is -CONHCHg COOH and R ⁸ is hydrogen This compound of formula 4 is known as FK-156

The novel immunostimulants of the present invention have the general formula I, wherein R5 is cycloalkyl with 4-7 carbon atoms, cycloalkylmethyl with 6-8 carbon atoms, or alkyl with 2-10 carbon atoms; Rg is hydrogen or alkyl of 1-3 carbon atoms and R3 is the residue of formula 2 of amino acid IIa where X is hydrogen, alkyl of 1-2 carbon atoms or hydroxymstyle, n is an integer from 0 to 4 and R ₄ and R are each independently hydrogen, alkyl of 1-6 carbon atoms, cycloalkylmethyl of 6-8 carbon atoms, or benzyl, and their pharmaceutically acceptable basic salts.

A preferred group of compounds of formula I are those in which R5 is alkyl of 5-8 carbon atoms, R2 is hydrogen, R3 is the residue of formula 2 of the amino acid of formula IIa, in which X, n, Rg are as defined above, and R4 represents a hydrogen atom. Especially preferred within this group are compounds of formula I in which n is zero and Rg is hydrogen, the above-mentioned alkyl or cyclohexylmethyl. Especially preferred compounds are those in which is / R, S [Beta] -2-ethyl-1-butyl, Rg is hydrogen and X is methyl; R 6 is (R), S (3-heptyl, R 6 is hydrogen and X is methyl; Rg is (R, S) 2-methyl-pentyl, Rg is hydrogen and X is methyl; R 4 is (R), S (2-heptyl, R 6 is hydrogen and X is methyl; R | is / R, S / 2-ethyl-1-pentyl, Rg is hydrogen and X is methyl; R 5 / R, S / 1-hexyl, R 8 is hydrogen and X is methyl, R 5 is / S / or / R, S / 2-methyl-1-haxyl, R 6 is hydrogen and X is methyl ; R 6 is (S) or (R ₁ S) 2-ethyl-1-hexyl, R 6 is hydrogen and X is methyl; and R5 is 1-hexyl,

Rg is hydrogen and X is methyl. Also especially preferred are compounds in which ^ 1 is 1-hsksyl, X is hydrogen and n-3. Especially preferred are the esters in which is (R, S) 2-atyl-1-pentyl, X is methyl and Rg is n-butyl, iso-butyl or cyclohexylmethyl: R4 is / S / or / R, S) 2-methyl-1-hexyl, X is methyl and Rg is n-butyl, isobutyl or cyclohexylmethyl and R5 is / S / or / R, S / 2-ethyl-1-hexyl, X is methyl and Rg is n-butyl, iso-butyl or cyclohexylmethyl

Another preferred group of compounds of formula 1 are those in which R5 is cycloelkylalkyl with 4-7 carbon atoms, Rg is hydrogen and R3 is the aforementioned residue of formula 2 of the amino acid of formula 2a, in which n 0, X is alkyl of 1-2 carbon atoms and R4 and Rg are each hydrogen. Particularly preferred within this group are those compounds in which R <2> is cyclohexyl and X is methyl.

A third preferred group of compounds of formula I are those in which R5 is alkyl with 5-8 carbon atoms, Rg is hydrogen, R3 is the aforementioned residue of the amino acid formula 2a, in which X is hydrogen or alkyl of 1 -2 carbon atoms, n is the integer 0-4, Rg is hydrogen and R4 is alkyl of 1-6 atoms, cycloalkylmethyl of 6-8 carbon atoms or benzyl

By pharmaceutically acceptable basic salts of compounds of formula I are meant salts with inorganic or organic bases such as metal hydroxides

150 055 alkaline or alkaline earths 1 ammonium, triethylamine, atanolamine 1 dicyclohexylamine · The configuration of the amino acid residues that make up the compounds of formula 1 It is significant in terms of the pharmacological activity of these compounds ·

The highest activity is observed in the compounds of formula 1, with the configuration indicated in formula U of those compounds of formula 1, in which R ₂ and X are different from hydrogen, the preferred stereochemistry at this carbon atom is L and D, respectively.

Methodologies for the preparation of compounds of formula 1 include the formation of peptide connections between amino acids which, due to their amino and carboxyl groups, and often due to the presence of other reactive groups, require the protection of these groups 1 / or the activation of such groups, especially carboxyl groups, to ensure the course of a specific reaction or for its optimization

Compounds of formula 1 according to the invention are prepared by coupling the passage of Formula 3 in which R, R ₂ and R ₄ are as defined above, with a fragment of the amino acid of formula 2a in which X n and R ₅ are as defined above ·

Some protecting and activating groups are illustrated in the following examples.

For specialists, it is obvious that other protecting and activating groups could be used. The choice of a specific protecting group largely depends on the availability of the necessary reagent, its effect on the solubility of the protected compound, the ease of its removal and the presence of other groups on which it can act, that is selectivity or its removal

For example, it may be necessary, or at least desirable in many reactions, to protect the amino group and / or the carboxyl group. The chosen route of peptide synthesis may require removal of one or the other or both of these protecting groups to ensure further reaction of the regenerated amino or carboxyl group is the protective groups are used are reversible, and in many cases they can be removed independently of each other.In addition, the choice of the protecting group for a given amino group depends on the role of this amino group in the entire course of the synthesis.It should be used for amino groups protective groups with different levels of lability, that is, the ease of their removal. · The same applies to protecting groups for carboxyl groups. · Such groups are known in the art and the following sources can be used: Badansky et al. · Peptide Synthesis 2-nd Ed. John Wiley and Sons, Ν · Υ · / 1976 /; Green Protective Groups in Organic Synthesis, John Wiley and Sons, N.Y. (1981); McOmie, Protective Grups in Organic Chemistry, Plenum Press, N.Y / 1973 /; and Sheppard, Comprehensive Organic Chemistry, The Synthesis and Reactions of Organic Compounds, Pergamon Press, N.Y. / 1979 /, edited by E. Hasiem, part 23.6, pp. 321-339. Conventional protecting groups for the protection of amino and carboxyl groups are known to those skilled in the art.

Representative, but not limited to, are the protecting groups for the amino group; benzyloxycarbonyl, substituted or unsubstituted aralkyl groups such as benzyl, trityl, benzhydryl and 4-nitrobenzyl; benzylidine, arylthio such as phenylthio, nitro-phenylthio and trichloro-phenylthio; phosphoryl derivatives such as dimethylphosphoryl and Ο, Ο-dibenzylphosphoryl, trialklysilyl derivatives such as trimethylsilyl and others, as described in U.S. Patent No. 4,322,341, which is hereby referenced. The preferred amino protecting group is benzyloxycarbonyl The process of substituting such groups in a given amino group is well known. Generally, it is acylating a certain amino group with benzyloxycarbonyl chloride / benzyl chloroformate / in a reaction-neutral solvent, e.g. water, methylene chloride, tetrahydrofuran in the presence of a base (acid acceptor), e.g. sodium or potassium hydroxide when the solvent is water, or an organic solvent is and is used in the presence of a tertiary amine such as C _1-4 -trialkylamine or pyridine. If an aqueous system is used, The pH of the reaction medium is kept in the range 8-10, preferably 9. Alternatively, if the reagent, i.e. the compound whose amino group is to be protected, has a basic group, it can serve as an acid acceptor.

The protected amino and carboxyl groups are converted into unprotected groups in a manner known to those skilled in the art. Benzyl group, a preferred protecting group for the carboxyl group / as part of the protected carbazoyl group / is removed by catalytic hydrogenation with palladium, mostly palladium on carbon. Alternatively, such groups are removed using

150 055 trifluoromethanesulfonic acid in trifluoroacetic acid »in the presence of enisol to prevent alkylation · The t-butoxycarbonyl group is easily removed by treatment with hydrogen chloride saturated dioxane ·

Activating a carboxyl group as a means to direct a given reaction is methodologically known to those skilled in the art to use anhydrides in the reaction sequence described below, especially cyclic anhydrides and activated esters such as those derived from N-hydroxyphthalimide and N-hydroxy succinimide, both of which are are used in peptide syntheses Activated N-hydroxy succinimide esters excellently direct the reactions at these activated ester groups It is known to those skilled in the art that other activating groups can also be used A particularly interesting group is the N-hydroxyphthalimide group, which is used in the same way as N-hydroxyimide groups of succinic acid · A dehydrating coupling agent is used in both cases to form the activated ester ·

Representatives of such coupling agents are; , diphenylketene and N-ethyl-5-phenylisoxazolene-3-sulfonate · The reaction conditions for the use of these coupling agents are widely described in the literature · A reaction-neutral solvent is generally used at room temperature to 100 ° C ·

The above-mentioned kerbodiimide reagents are particularly recommended because they allow the reaction to be carried out at ambient temperature and allow a satisfactory yield of the desired ester to be obtained.

After the coupling reaction to the final product is complete, the various foot protecting groups are removed by the methods previously described and the compounds of formula 1 are isolated.

The pharmaceutically acceptable salts of compounds of formula I in which R5 or Rg are hydrogen are obtained by treating a solution, preferably an aqueous solution of such a compound of formula I with a base as mentioned above, usually in stoichiometric proportions. The salts can be isolated by evaporation or by precipitation ·

The products produced according to the invention will be processed into pharmaceutical compositions in unit doses containing a pharmaceutically acceptable carrier and an effective amount of a compound of formula 1 with an anti-inflammatory or immunostimulating effect.

The products produced according to the invention are useful for the treatment of mammals, including humans, both clinically and prophylactically in diseases caused by various pathogenic microorganisms, especially gram-negative bacteria. They are also useful as immunostimulants in mammals, including humans, with a high risk of infection due to the existence or clinically induced immunodeficiency

A test study using male C ^ H / HeN mice from Charles River Breeding Laboratory is shown below

Mice are acclimated for 5 days prior to use, and then dosed in a volume of 0.2 ml subcutaneously / SC / or orally / PO / test compounds at various dilutions / 100,

10.1 and 0.1 mg / kg / or placebo / pyrogen-free saline / The regimen of action depends on the organisms used for infection: -24 and 0 hours before inoculation with Klebsielle pneumoniae in normal and -3, -2 and - mice 1 day before inoculation with Escherichla coli or Staph, aureus in immunocompromised mice The infection is carried out intramuscularly / IM / in the thigh in the case of K pneumoniae or intraperitoneally / IP / in the case of E. coli and Staph aureus. , 2 ml Mortality is recorded after 7 days for K pneumoniae and after 3 days for the other two micro-organisms

Cultures of K pneumoniae, E coli 1 Stach aureus from a frozen blood bank for purity were carried out on infusion cerebral nerve agar / BHI /

From 18-hour-old culture plates, three colonies were inoculated onto 9 ml of BHI broth. The culture was grown for 2 hours at 37 ° C in a rotary shaker, then 0.2 ml was applied to a series of BHI slants. After 18-hour incubation at 37 ° C C slants were washed with BHI broth, culture density was corrected using Spectronić 20 and diluted to obtain LO5O dose for normal mice.

150 055

When used as anti-inflammatory or immunostimulating agents in humans, the compounds of the invention are administered orally, subcutaneously, intramuscularly, intravenously or intraperitoneally, generally in the form of a composition according to pharmaceutical practice. For example, they may be administered in the form of tablets, pills, powders or granules. , containing excipients such as starch, milk sugar, some types of clays, etc. They can be administered in the form of capsules mixed with the same or equivalent excipients They can also be administered in the form of oral suspensions, solutions, emulsions, syrups and elixirs that can contain flavoring or coloring agents.For most oral applications, tablets or capsules containing 50 to 500 g of the substances according to the invention are best suited.

The physician should determine the most effective dosage for each patient, depending on age, body weight, response and route of administration. Oral administration is recommended in single or divided doses in the amount of 1.0 to 300 mg / kg / day. Recommended dose for parenteral use - is 1.0 to 100 mg / kg / day, preferably 1 to 20 mg / kg / day

The following examples are provided solely to better illustrate the invention. The following abbreviations are used in the NMR spectra descriptions: s - slnglet, d - doublet, t-triplet, q - quartet, m - multiplet · Meaning moles 1 millimoles are defined by the abbreviations m and ma

Example I · N-heptanoyl-D-gamma-glutamyl-gllcylo-D-alanlna / R-Chg / ^{CH 2/5 '1 R} 2 ^H + ^{and R} 3' - ^hndch / C ^H 3 / C ⁰ 2 ^h / ·

IA. N-heptamoyl-D-gamma-gluteaylo / benzyl alpha ester / -glycine

To a solution of 897 mg / 13 mm / glycine and 1.3 g / 13.0 mm / triethylamine in 10 µl of water was added 5.0 g (11.2 mm) of the hydroxy succinimide ester of N-heptanol-O-gamma-glutamylate elf benzyl in 100 ml of dioxane, and the mixture thus formed was stirred at room temperature for 80 hours. The solution was poured into 300 ml of ethyl acetate and the separated organic phase was washed with 10% hydrochloric acid, water and brine solution. The organic phase was separated, dried over magnesium sulphate and evaporated to dryness under reduced pressure. The residue was triturated with diethyl ether and filtered under nitrogen. yielding 3.43 g / 74% / product yield

IB. N-heptanoyl-O-gamma-glutamyl-glycyl-D-alanine

For a solution of 2.0 g / 4.78 mm / N-haptenoyl-D-garara-glutamyl / alpha-benzyl ester / -glycine, 1.75 g / 5 mm / p-toluenesulfonic acid salt of D-alanine benzyl ester, 506 mg (5 mm) of triethylamine and 675 mg (5 mm) of i-Nydroxybenzotriazole in 100 ml of tetrahydrofuran, 3.03 g (7.17 mm) of 1-cyclohexyl-3- (2-morpholinoethyl) -carbodiimide metho p-toluenesulfonate were added and so on the resulting mixture was stirred at room temperature for 18 hours. The reaction mixture was poured into 300 ml of ethyl acetate, the organic phase was separated and washed with 10% hydrochloric acid, water, saturated sodium bicarbonate solution and brine solution. The organic phase was separated, dried over magnesium sulfate and concentrated under reduced pressure. The residue was triturated with diethyl ether and filtered under nitrogen to give 2.7 g of the product. Two grams of this solid in 75 ml of methanol with 400 mg of 10% palladium on carbon was shaken under an atmosphere of hydrogen at an initial pressure of 0.345 MPa for 4 hours.

The catalyst was filtered off, the filtrate was evaporated under reduced pressure and the residue was dissolved in water and lyophilized to give 1.23 g (90% yield) of the desired product as a white solid.

NMR spectrum (DMSO-d 6) showed absorption at 4.35-4.2 (m, 2H), 3.83 (s, 2H), 2.35 (t, 3-7 Hz, 2H), 2.17 (t, 0 - 7 Hz, 2H), 2.1-1.8 (m, 2H), 1.55-1.45 / m, 2V, 1.3 (d, J = 6ffe, 3H), 1. 17 (bs, 6H) and 0.75 (ba, 3H), ppm.

EXAMPLE II · N-heptanoyl-D-gamma-glutamyl-glycyl-glycine / R "CH ₃ / CH _2/5 -jr -hj ₂ r _{3 2} -hnch every ₂ h /

IIA · N-heptanoyl-D-tetra-glutamyl hydroxy succinimide ester / benzyl alpha ester / -glycine.

For a cold solution / 0 ° C / 13.0 g / 31 mm / N-heptanoyl-D-gamma-olutamyl / alpha benzyl ester / -glycine and 3.91 g / 34 mm / N-hydroxy succinimide in 400 ml tetra6

150,055 of hydrofuran, 7.0 g (34 mm) of dicyclohexylcarbodimide was added and the mixture was stirred at 0 ° C for 1 hour and then at room temperature for 18 hours.

The solids were filtered off and the filtrate was evaporated under reduced pressure. The residue was triturated with diethyl ether and filtered under nitrogen to provide 15.4 g (98%) of the desired product.

IIB. N-heptanoyl-D-gamma-glutemyl-glycine

To 2.0 g (3.97 mm) of N-heptanoyl-D-gamaglutamyl hydroxy succinimide ester / benzyl / -glycine alpha ester in 100 ml of dioxane 446 mg (5.96 mm) of glycine and 0.55 ml / 3 were added , 9 mm / triethylamine in 10 ml of water and the resulting mixture was stirred at room temperature for 18 hours. The solution was poured into 100 ml of ethyl acetate and the organic layer was washed with 2.5% hydrochloric acid, water and brine solution. The organic layer was separated, dried over magnesium sulfate and evaporated to dryness. The residue was triturated with diethyl ether and filtered under nitrogen to give 1.7 g of a white solid. 1.5 g of this solid in 75 ml of methanol containing 200 mg of 10% palladium hydroxide on carbon was shaken under a hydrogen atmosphere at a pressure of 0.345 MPa for 3 hours. The catalyst was filtered off and the filtrate was concentrated under reduced pressure. The residue was dissolved in water and lyophilized to give 1.12 g (90% yield) of the desired product.

NMR spectrum (DMSO-d6) showed an absorption at 8.2-8.0 (m, 3H), 4.19 (m, 1H).

4.8-4.6 (, 4H). 2.25 (t, 0 - 7 Hz, 2H), 2.1 (t, 0 - 6 Hz, 2H), 2.05-1.7 (m, 2H), 1.5 (m, 2H), 1, 25 / be, 6H / 1 0.85 / t, 0-6Hz, 3H / ppm.

Example III. N-heptanoyl-D-gamma-glutamyl-glycyl-D-9erine (R 1 »CH ₃ / CH ₂ ) _{5 -t} R 2 — hn 8h (CH ₂ oh) (every ₂ h).

Starting from 2.0 g / 3.98 mm / N-heptanoyl D-gamma-glutamyl hydroxy succinimide ester / benzyl alpha ester / -glycine, 780 mg / 4.02 mm / O-benzyl-D-serine and 0.556 ml / 4.02 mm (triethylamine) and proceeding as in Example 2B to give 902 mg (76% yield) of the desired product, mp 130-132 ° C.

NMR spectrum (DMSO-d_) showed absorption at: 8.36-7.94 (m, 3H), 4.46-4.28 (m, 1H),

4.28 - 4.08 /, 1H /, 3.94 - 3.50 / m, 4H /, 2.25 / t, 0-9Hz, 2H /, 2.17 / t, J-9Hz, 1H / , 2.10-1.04 (, 14H) and 0.9 (t, 0-6Hz, 3H) ppm.

Example IV. N-O-heptanoyl-gamma-glutamyl-glycyl-D-alpha-amino-butyric acid / R ₁ CH ₃ / CH _2/5 -> R-H, R ₅ '-HNCD / CH ₂ CH ₃ / CO ₂ H /.

The procedure of Example 2B was repeated, starting with 2.0 g (3.98 mm) of N-heptanoyl-D-gamma-glutamyl hydroxy succinimide ester (alpha benzyl acid) -glycine,

400 mg (4.02 mm) of D-alpha-amino butyric acid and 0.556 ml (4.02 mm) of triethylamine afforded 632 mg (57% yield) of the desired product, mp 140-141 ° C.

NMR spectrum (DMSO-d6) showed absorption at: 8.16-8.04 (m, 3H), 4.22-4.08 (m, 2H), 3.84-3.58 (", 2H). 2.2 (t, 3 = 9 Hz, 2H), 2.12 (t, J? 9 Hz, 2H), 2.04-1.0 (m, 15H) and 0.85 (m). 0 · 6Ηζ, 6H / ppm.

EXAMPLE acid N-heptanoyl-D-gamma-glutamyl-glycyl-3-aminopropionic acid / r ₁ CH ₃ / CH _2/5 - r h _2, r ₃ -HN / CH _2/2 - at ₂ h /.

The procedure of Example 2B was repeated and starting with 1.5 g (3.0 mm) of N-heptanoyl-O-gamma-glutamyl hydroxy succinimide ester / alpha benzyl ester-glycine, 350 mg (3.9 mm) of 3- of aminopropionic acid and 0.56 ml (3.9 mm) of triethylamine, 500 mg (43% yield) of the desired product were obtained, mp 135-138 ° C.

NMR spectrum (DMSO-d6) showed absorption at: 8.19-8.02 (m, 2H), 7.98-7.87 (t,

0-5Hz, 1H), 4.25-4.1 (m, 2H), 3.8-3.49 (m, 2H), 3.44-3.1 (m, 2H), 2.4) t, 0-6Hz, 2H / 2.22 / t, 0-7Hz, 2H /, 2.14 / t, J-7Hz, 2H /, 2.1-1.67 / m, 2H /, 1.6 -1.17 (m, 8H) and 0.88 (t, 0-6Hz, 3H / ppm.

Example VI. N-heptanoyl-D-gamma-glutemylo-glycyl-4-aminobutyric acid / y ₁ ^'CH 3 / ^CH _2/5 -, R ₂ "H, R ₃ -HN / CH _2/3 CO ₂ H /.

150 055

The procedure of Example 5 was repeated, replacing 4-aminopropionic acid with 410 mg (4.0 mm) 4-amino butyric acid to give 600 mg (50% yield) of the desired product, mp 140-142 ° C.

NMR spectrum (DMSO-d ₆ ) showed absorption at: 8.18-8.03 (m, 2H), 7.88 (bt.

0 -4Hz, iH), 4.17-4.09 (m, 2H), 3.81-3.48 (m, 2H), 2.32-2.08 (m, 6H), 2.08- 1.08 (m, 12H) and 0.88 (t, J = 6 Hz, 3H) ppm.

Example VII. N-heptanoyl-D-gamma-glutamyl-glycyl-5-aminopentanoic / R CHIg / Cl · ^ / ^ -, R ₂ ^a H, R3 »-HN / CH _2/4 C0 ₂ H / ·

Replacing 3-aminopropionic acid with 470 mg (4.0 mm) of 5-amino-pentanoic acid by following Example 5, 520 mg of the desired product were obtained, m.p. 22-124 ° C.

NMR spectrum (DMSO-d6) showed absorption at: 8.25-7.94 (m, 2H), 7.85 (t, □ -5Hz, IH), 4.25-4.1 (m, 2H), 3.82-3.45 (m, 2H), 3.24-2.9 (m, 2H), 2.21-2.08 (m, 6H),

2.08-1.2 (m, 14H) and 0.88 (t, 0-6 Hz, 3H / ppm.

Example VIII. N-heptanoyl-D-gamma-glutamyl-glycyl-6-amino-hexanoic acid / CH ^ R ^ / CH ^ / ^ -, R2 ^eH "R ^ -HN / CH ^^ COgH /.

Substituting 3-aminopropionic acid with 530 mg (4.0 mm) of 6-aminohexanoic acid and proceeding according to Example 5, 520 mg (40% yield) of the desired product were obtained as a white foam.

NMR spectrum (DMSO-d _g ) showed absorption at: 8.28-7.9 (m, 2H), 7.82 (bt)

0 "4Hz, IH), 4.27-4.1 (m, 2H), 3.81-3.47 (m, 2H), 3.15-2.90 (m, 2H), 2.3- 2.08 (m, 6H), 2.08-1.18 (m, 16H) and 0.88 (t, 3.6Ηζ, 3H) ppm.

Claims (6)

Patent claims A method for the preparation of new acylpeptides of the general formula, in which R5 is alkyl of 2-10 carbon atoms, cycloalkyl of 4-7 carbon atoms or cycloalkylmethyl of 6-8 carbon atoms, R2 is hydrogen or alkyl of 1-3 carbon atoms, R3 represents the remainder of the formula 2a of the amino acids of formula IIa, wherein X is hydrogen, alkyl of 1-2 carbon atoms or hydroxy methyl, and n is an integer of 0-4, and R4 and R5 each independently represent hydrogen or pharmaceutically acceptable basic salts thereof, characterized by the compound of formula 3 wherein and R ₂ are as defined above is coupled with the simultaneous dehydration of a compound of formula 2a, wherein X, n and R are as defined above, and the groups are not included in the reaction are optionally protected, and selectively removing the blocking groups and, if desired, converting the compound of Formula 1 to its pharmaceutically acceptable base salt. 2. The method according to claim 1, characterized in that the coupling is carried out with N-hydroxy succinic acid or 1-hydroxybenzotriazole and a carbodiimide. The method of Claim 1, wherein the O-benzyl blocking groups are removed by catalytic hydrogenation. 4. Process for the preparation of new acylpeptides of general formula I, wherein R5 is alkyl of 2-10 carbon atoms, cycloalkyl of 4-7 carbon atoms or cycloalkylmethyl of 6-8 carbon atoms, Rg is hydrogen or alkyl of 1-3 carbon atoms, R3 is a residue of formula 2 of the amino acid of formula IIa, where X is hydrogen, alkyl of 1-2 carbon atoms or hydroxymethyl, and n is an integer of 0-4, and R4 and Rg are each independently alkyl of 1- 6 carbon atoms, cycloalkylmethyl of 6-8 carbon atoms or benzyl, or if R4 or Rg is hydrogen, the latter is as defined above and pharmaceutically acceptable base salts thereof, characterized in that the compound of formula III, wherein R 2e, where X, n and Rg are as defined above, and the non-reacting groups are optionally β 150 065 unprotected, the blocking groups are then selectively removed and the compound of Formula 1 converted to its pharmaceutically acceptable basic salt if desired. The method according to claim 4, characterized in that the coupling is carried out with N-hydroxy succinamide or 1-hydroxybenzotriazole and carbodia midium The method according to claim 4, characterized in that the O-benzyl blocking groups are removed by catalytic hydrogenation. ABOUT RiĆNH D / CO2R4 CH L O I II (CH ₂ ) ₂ CONHCHC-R ₃ R ₂ Formula 1. D -NHeH (CH ₂ ) _n CO ₂ R ₅ X Formula 2 D HiNCHCHzLCCbRs X Formula 2o ABOUT RiĆNH D _z CO ₂ R ₄ Yh L ICH ^ mNHCHCO ^ 'Formula 3 R2 R ¹ - (HNCHCO) n-HN-CH-R ³ R ² (C Hzlm CO-NH-CH-R '* (CH ₂ ) ₃ Formula 4 R ⁶ -HN-ĆN-R ⁵ Printing House of the Polish Patent Office. Circulation 100 copies Price 1500 PLN