SU639446A3 - Method of obtaining peptides - Google Patents

Description

where R2 and R 1 have the above values, followed by the removal of all remaining hydrochloric acid groups. Example 1. Trifluoroacetate of alanyl-hdistil methyl ester (H-Ala-Gly- -OMe. CFsCOOH). 10 g of BOC-Ala-G y-OMe (45 mmol} is dissolved in 100 ml of cold (ice on a bath) of trifluorosulphonic acid. The solution is stirred for 1 hour at 0 ° C and the solution is distilled off (solvent, the residue is dissolved in methanol, Diethyl ether is added and the precipitated product is filtered off. EXAMPLE 2 Tert methyl ester, butyl oxy-benyl-glycyl-ala-glycine (BOC-Gly-Ala-Gly-OMe). To cold (ice on a ban) a solution of H-L1a (-Gly-OMe-CH3COOH (45 moles) in d} 1., 1 methyl ethyl (50 w)) is added. -this, lMorfolin (pH 8), then a cooled solution of 18 g of BOC- Gly-Otsr (45 lsheds in dim € ti, lfor moms Ideal (50%). The solution is kept on an ice bath for 2 days. The solvent is distilled off, the product is purified by chromatography on a column of silica gel using ethyl acetate (methanol) pyridine (98: 1: 1) as an eluent. The product is crystallized from a mixture of ethyl acetate and petroleum ether, the above compound is obtained, mp 98-100 ° C, NMR (DMSO-de); 1.25 b (3N), 1.4 b (9P), 3.68 b ( ZN). Example 3. Trifluoroacetate methyl ester gly | Tsil-alanyl-gly1 tsina (H-Gly-α-Ala-Gly-OMe. NUAF). 6.4 g (20.1 mmol) of BOC-Gly-Ala-iGly-OMe are dissolved in cold (ice-bath) trifluoro-x-muonic acid (120 ml) and the solution is stirred at 0 ° C for 1 hour. The solvent is distilled off, the residue is p, dissolved in methanol, the product is precipitated by addition of diethyl ether and filtered. Example 4: M-ester Grete, butyloxycarbonyl - glycyl - glycyl-alanylglyce .in (BOC-Gly-Gly-Ala - Gly - -OMe). To cold (ice Na-ban) solution 6.4. g (20.03 mt) H-Gly-Ala-Gly- -OMe GHsCOOH in dimethylformamide (30 ml) N-ethylmorpholine (2.8 ml} is added with stirring, then a solution of 8.5 g (24 mmol) of BFB- Gly-Otor in dimethyl form and MFA (20 ml. The solution is kept on ice-cold 1bahn, 2 days. The solvent is distilled off, the residue is dissolved in methanol and the product wasps; wait for the ethyl ether, crystallized from ethyl acetate, the above compound is obtained, mp 103–105 ° C and 141 ° C (dimorphic); NMR 1.25 5 (3N), 1.38 b (9H), 3.65 b (3N). Example 5. Hydrazide treg. butyloxycarbonyl- GLITNGLYCYL - ALAL - Glycine (BOC-Gly Gly-Ala - Gly - NH - -NH2). To the cold ( ice on a bath) to a solution of 2.5 g (6.7 mmol) of BOC-Gly-Gly-Ala-Gly-OMe in methanol (50 ml), 2.5 ml of shdrazine hydrate is added with stirring. The solution is stirred at 0 ° for three hours and at a constant temperature overnight. The precipitate is filtered, washed with methanol, dried, the above compound is obtained; amino acid analysis: Gly 3; Ala 0.99. Example 6. Greg butyloxycarbonyl methyl glycyl glycyl glycine allyl glycyl C-trityl methyl ester -cysteinyl-N-G; og. butyloxycarbonyl-lysyl - asparagine - phenylalanyl-phenylalanine. (BOC-Gly-Gly- -Ala - Gly - Cys (Trit) -Lys- (BOC) - -Asn-Phe-Phe-OMe). To a solution of 1.57 g (4.2 mmol) of BOC-Gly-GLy-Ala-Gly NH-NHj in dimethyl sulfoxide (25 ml) and dimethyl farmide (25 ml) were added 2.04N hydrogen chloride solution in ethyl acetate (5.15 l) at -10 ° C. The mixture is cooled to -12 ° C and 0.61 ml (5.2 mmol} of greg. butyl nitrite is added. It is kept at C 15 min, O1 is cooled to -15 ° C and 1.8 ml (pH 8) of N-ethyldiisopropylamine, then a solution of 4.5 g (4.18 mmol} H-Cys (Trit) -Lys (BOC) -Asn-Phe- -Phe- OMe and again 0.72 ml of N-ethyldiisopropylamine in dimethylformamide (25 ml}. The mixture is stirred for one hour at -10 ° C and overnight at room temperature It is evaporated, the residue is dissolved in methanol, the product is precipitated with water and crystallized from methanol to obtain the above compound; aiMiH but acid analysis: Lys 1.07; Asn 0.97; Gly 3.27; Ala 1.0; Phe 2.08 cysteic acid 1.35. Example 7. Gret.butyloxy .carbonyl-glvcyl-tlicyl-alanyl - glycyl-Strityl-cysteinyl-N - treg.butyloxycarbonyl-lysyl - asparaginyl - phenylalanine - phenyl alanine hydrazide. (BOC-Giy-Gly-Ala-Gly- - Cys (Trit) -Lys (BOC) -Asn-Phe-Phe- -NHNHa). To a cold (ice-bath) solution of 0.9 g (0.66 mol} BOC-Gly- -Gly-Ala-Gly-Cys (Trit) -Lys (BOC) - -Asn-Phe-Phe OMe in dimethylformamide (25 ml) 1.5 ml of hydrazine hydrate is added. The solution is stirred for 18 hours at room temperature. The solvent is distilled off, the residue is triturated with methanol and dried over phosphorus pentoxide, but the above compound; Amino acid analysis: Lys -1,10; Asn 1,00 ; Gly 3,33; Ala 1,0; Phe 2,14; cysteic acid 1.35. Example 8. Greg. Butyloxycarbonylgly: Cytil-glycyl-1-alanyl-1 Gly Tsil-8-trityl-stiine-N-Greg. Butyloxycarbonyl- lysylasparaginyl - phenylalanil - phenyl laniltryptophil-N - Greg.butyloxycarbonyl-lysyl-0-treg. butyl-threonyl - fepylanyl-Ogreg. butyl-threonyl - phenyl) 1nl - O-tert, butyl-threonyl-0-gret. butyl - seril - S-trityl cysteine . BOC-G y-GlV-Ala-Gly-Cys (Trit) -Lys (BOG) -Asn-Phe-Phe- -Trp - Lys (BOG) -Thr (Bu) -Phe- -Thr (Bu) -Ser ( Bu) -Gys (Trit) OH.

To a solution of 800 mg (0.59 mmol) BOG- -Gly-Gly-Ala-Gly-Gys (Trit) -Lys- (BOG) -Asn-Phe-Phe-NHNHj in dimethylformamide (12 ml, cooled to -20 ° C, 1.85 g of a solution of hydrogen chloride in egylacetate (0.795 ml, 1.475 mmol) is added. GMecb is cooled to G and 0.081 ml (0.71 mmol) of tert-butyl nitrite is added, the solution is mixed with 15 ml. The solution is 81Q ml (0.59 ml) ) H-Trp-Lys- (BOG) -Thr (Bu) - Phe - Thr (Bu) - Ser- (Bu) -Cys (Trit) -OH and 0.354 ml (2.06 mmol) of N-ethyl-diisopropylamine in dimetklformamide ( 6 ml) is cooled to -15 ° C and added to the reaction mixture, which is stirred at -15 ° G for 1 hour and at 25 ° G for 18 hours. The solvent is distilled off, the residue triturated ayut with icy citric acid, filtered, washed with water and methanol, dried, the above compound is obtained; amino acid analysis: Zys

BOC-G-lij-G-lij-Aia-Gly-Yi) 5-Lijg BOC Agr.-he-Ph.e-Trp-LijS (BOCl-Th.) - Phe-ThrCBu) -ger (Bu) -Cy5 -c; i

To this compound is added cold concentrated hydrochloric acid (23 ml) in an ice-water bath under nitrogen with vigorous stirring for 10 minutes, then up to: add glacial acetic acid (300 ml) and the solution is lyophilized. . The residue was dissolved in water and lyophilized again, the residue was dissolved in 0.01 n. aqueous solution of mmonium acetate and added to the column with carboxymethylcellulose (Whatman GM-23, 2.5X30 cm). Products elute 0.06 n. ammonium additate solution (buffer). The purified substance is lyophilized and the above compound is obtained in the form of a white precipitate in the form of its acetic salt; Ltah MeOH 282 microns (to 5260), 289 microns (e 4730): amino acid analysis: Lys 2.28; Asn a, 05; Thr 1.95, Ser 0.98. Gys 2.34, Gly 3.00, Ala 1.05, Phe 3.12. Newly lyophilized compound gives the compound as a free base; amino acid analysis: Lys 2,10; Asn 1.04; The 1.80; Ser 0.95; Gvs 2.17; Gly 3.00; Ala 1.07; Phe 2.99.

Similarly, you can get the above compound, Lrimen instead of iodine Rodan: according to the method of Hiski. And G.MHTa.

Example 10. Methyl ether treg.butyloxycarboyl. Nyl-leu | cyl - glycyl-glycyl1, 04; Asn 0.96; Thr 1.6; Ser 0.49; Gly 2.82;

Ala 1.0: 0; Phe 2.80.

lO p and M e p 9. Cyclic disulfide glycyl-glycyl-alanyl-glycyl-cysteinylisyl-asparaginyl-phenylalanyl-phenyl-nnl-Triitophil-lysyl-threonyl-phenyl-aryl-threonyl-seryl-cysteine. (H-Gly-- -Gly-Ala-Gly-Gys-Lys-Asn - Phe- -Phe-Trp-Lys-Thr - Phe - Thr-Ser - -Gys-OH).

0.871 g (0.32 mmol) BOG-Gly-Gly- -Ala-Gly-Gys (Trit) -Lys (BOG) -Asn- -Phe-Phe-Trp-Lys (BOG) -Thr (Bu) - -Phe -Thr (Bu) -Ser (Bu) -Gys (Trit) -OH is dissolved in glacial accelerated acid (60. "l) and 150 ml of iodine in methanol is added dropwise to a 0.5% HOMV solution of iodine in (30 mmol) with stirring for 1 hour at room temperature. GMecb is stirred for a further e for 45 minutes, then cooled on ice and 1 N added. a solution of 6 ml of sodium hyposulfite in water to destroy excess iodine (colorless solution). The solvent is distilled off, the residue is triturated with water, dried and the dry product is triturated with isopropyl ether, to obtain a cyclic disulfide of hexadecanapeptide of the formula

alanyl-glktsia (BOG - Leu-Gly-Gly- -Ala-Giy-OMe).

2 g (5.35 m.iol) of a BOG-Gly-α-Gly-Ala-Gly-OAVe solution in triftoracetic acid (25 ml) was stirred at 0 ° G for one hour. The solution of the intel is distilled off, the residue is triturated with e-f1f, the residue is ground and dried, and the tetrales of the bird H-Gly-Giy-Ala-Giy-OMe are obtained as a trifluoroacetic salt. 5.35 mmol of the peptide solution, 2.31 g {lu mlol) BOG-Leu-OH 1.35 g m-, 10 l) lozbis triazole, 2.27. (11 ml) dicyclo.ohexylcarbodimide and 0.68 g of N-et.ilmorpholine from dimethylphoramide (25 ml permixed at 0 ° G for 24 hours. The precipitate is filtered off, f) glt. Solution is added to diethyl ether (200 ml). The precipitated precipitate separates ji crystal 1zuig from a mixture of methanol and isopropyl ether to obtain the above compound; so nl

V1 p) o / g

(G 1, dime 190, 5-193 ° G,

-10, tilformamide).

Example 11. Gret.butyloxycarbonyl-leucyl - glycyl - glycyl-alanyl-glycine hydrazide. (BOG-Leu-Giy-Gly-Ala-Gly-NH NHa).

2 g (4 liy.) Solution of BOG-Leu- -Gly-Gly-Ala-Giy-OMe n 4.12 ml (80 mmol) of hydrazine hydrate in methanol

(50 ml) stirred at 0 ° C for 4 h. The mixture is concentrated} by 1pairing to 4 ml and added to diethyl ether (200 ml). The precipitate was collected, dissolved in methanol (4 ml) and added to diethyl ether (200 ml). The precipitated precipitates are separated, dried and the above compound is obtained, i.e. 174-176.5 ° C; and f -12.0 ° (with I, dimethyl, or.ma; mi1d).

Example 12. Methyl ef.ir tert.butyloxycarboiyl-leucyl-glycyl-glycyl-alanyl-glycyl-S-tritnyl-cy-teinyl-N Greg. Butyloxycarbonyl-lysyl-asyaragi nyl-phenylalan. -Leu-Ghi-Gly-Ala-Gly-Cys (Trit) - Lys (BOG) -Asn-Phe-Phe-OMe).

0.15 ML (1.27 mmol) of tert. Buttons are added to a solution of 0.308 g (0.635 mmol) of BOC-Leu-Gly-Gly-Ala-Gly-NsHs in dimethylformamide (5 ml) at -20 ° C and 2.4 and. - hydrogen chloride in ethyl acetate (0.66 ml, 1.59 mmol). Stir 15 min. three -90 ° G to a mixture add a solution of 0.62 g (0.58 mmol) of H-Cys (Trit) -Lys (BOG) -Asn-Phe-Phe-OMe and 0.372 ml of diisopropylethylamine in dimethyl form; mid (6 ml) The mixture is stirred at -20 ° G for one hour and at 0 ° G for 24 hours, then added to 200 ml of diethyl ether. The precipitated precipitate was collected, dissolved in methanol (5 liters) and added to diethyl (Ether (200 ml). The precipitate was filtered off and dried, and the above compound was obtained in T. 238-240.5 ° G.

Example 13. Greg. Gly-Gys (Trit) -Lys- (BOG) -Asn-Phe-Phe-, NHNH2).

A solution of 0.56 g (0.38 mmol) BOG- -Leu - Gly - Gly - Ala - Gly-Gys- - (Trit) -Lys (BOG) -Asn - Phe-Phe - -OMe and 0.74 ml of hydrate hydrazine in dimethylformamide (10 ml) was stirred at 0 ° G for 12 hours and three .25 ° G for 24 hours. The solution is added to 100 ml of diethyl ether, you (H-Leu-Gly-Gl j-Ala Solution 0.23 g (0.081 m.iol) BOG-Leu-Gly-Gly-Ala-Gly-Gys (Trit) - Lys (BOG) -Asn - Phe-Phe - Trp - - Lys (BOG) - Thr (Bu) -Phe - Thr (BuO-Ser (Bu) -Gys (Trit) -OH in acetic acid (90 ml)) drop by drop m for one hour is added to a solution of 0.5% iodine in methanol (41.5 ml, 0.81 mmol). The precipitated precipitate is collected, dissolved in dimethylformamide (3 ml) and added to 100 ml of diethyl ether. The precipitate is collected, dried and the above compound is obtained, mp 239-242 ° G; amino acid analysis: Lvs 1.05; cysteic acid 0.66; Asn 0.99; Gly 3.00; 1.11; 0.5 Gys 0 , 21; Leu 0.96; Phe 1.86.

Example 14. Greg.butyloxycarbonylleucyl - g LI1Ts and l-gl search for l-a.la and l - glycyl-S-trityl-cietenyl-№-greg.butyloxycarbonyl-lysyl-asyaraginyl - phenyl-aryl-phenyl-aryl-triitophil - N-trgt. butyloxycarbonyl-lysyl-O-g / et. butyl - threoyl-phenylalanil-0-g / er. butyl - threonyl-0 - tert. butyl-seryl-3-tert-cysteine (BOG-Leu-Gly-Gly-Ala-Gly-iCys (Trit) -Lys (BOG) -Asn-Phe-Phe-Trp-Lys- (BOG) -Thr ( Bu) -Phe-Thr (Bu) -Ser- (BuO-Cys (Trit) -OH).

0.03 ML (0.28 mmol) of greg.-butyl nitrite is added to a solution of 0.20 g (0.14 mmol} BOG-Leu-Gly-Gly-Ala-Gly-Gys (Trit) -Lys (BOG) -Asn -Phe-Phe-NsHs in dimethyl sulfoxide (2 ml) and dimethylformamide (4 ml) and 2.21 N. of hydrogen chloride in ethyl acetate 0.16 ml (0.35 mmol) cooled to -20 ° G. 1 at -20 ° G, a solution of 0.20 g (0.14 mmol) of H-Trr-Lys (BOG) - -Thr (Bu) -Phe-Thr (Bu) -Ser (Bu) Gys (Trit) - is added on-HGOOH and 0.08 ml (0.49 mmol) of diisopropylamine in dimethylformadide (5 ml). The solution was stirred at -20 ° G for 1 hour and at 0 ° G for 24 hours. Then concentrated to 2 ml and added to d. ethyl ether (100 ml), the precipitate is collected, washed one (2x5 ml), methanol (2x5 ml) and dried, the above compound is obtained; amino acid analysis: Zys 1.92; disteic acid 0.93; Asp 0.93; Thr 1.38; Ser 0.66; Gly 3 , 00; Ala 1.02; Gys 0.39; Leu 0.93; Phe 2.70.

Example 15. Cyclic disulfide leucylglycyl-tlitsil - alanyl - glycylCysteinyl-lysyl-asparaginyl - phenyl-rshl-feiyllail-tri-tofil - lysyl-threo yl-phenylalanyl-treoiyl-seryl-disteina

The solution is stirred at room temperature for 1 hour and cooled to 0 ° G. Introduce 1.62 ml of sodium hyposulfite until the solution completely discolored. The solvent is distilled off, the precipitate is separated, dried,

triturated with ether, dried and crystallized heptadecapepeptide cyclic disulfide of the formula Glij-Cys-Lys-Asn-Phe-P.he-TrpLL 3-Thr-Phe-Thr-Ser - (: t) S-0} -))

BOC-Glu-G-ly-A a-Gly-Cys-Lys (BO (;) - ASn-Phe-Phe-Trp-LyS (BOCV

-Thr (Bu) -Phe Tlvr№u) -Ser (Bu) -Cy & -OH

A solution of 0.155 g (0.066 zlnol} of heptadekapeptide peptic diculfide disulfide in concentrated hydrochloric acid (6.87 ml) under nitrogen atmosphere is vigorously stirred at 0 ° C min., Acetic acid is added (69 ml and lyophilized. Residue; create in 1 ml water (50 ml and again lyophilized, the product is chromatographed in a column of ic chemically modified with Sephadex G-25 M (3X60 hedgehog) interlaced dextran and equilibrated in the fusible phase, then in the upper phase of the butanol: water: 4: 1: 5 mixture .The upper phase of the LD; l: eny to desorb the peptide. The fracters containing the pure peptide are collected , evaporated and lyophilized from water to give the above in the form connect .nie th its acetate salt.

UV-proof, Yatach (MeOH): 290 lsh (e5000), 281 lc (E 5540), 275 microns (B 5205), 269 m (e 4980), 265 microns (e 4625), 259 lc. (e 4085). Repeated lyophilization of this product from water gives the above compound as its free base; amino acid analysis: Lys 1.98; cystennova acid 1.41; Asp 1.26; Thr 1.92; Ser 96; Gly 3.00; Ala 1.01; Leu 0.96; Phe 2.76.

Similarly -moz about get the above compound, applied instead of iodine filed.

Example 16. Methyl ether treg.butyloxycarbonyl - tlycyl-glycyl - glycylalanyl-glyshchina (BOC - Giy - Gly - Gly - Ala - Gly - OMe).

A solution of 3.7 g (10.04 l10.l) BOC-Gly-Otosr, 3.12 g (8.03 mmol} H-Gly-Gly-Ala-Gly-OMe. CP3 COOH and 1.1 ml morpholine in di: methylformamide (15 ml} leremeshivall at 0 ° C for 20 hours. The precipitate is filtered off, the filtrate is added to diethyl ether. The precipitate is separated, combined with the previous one and crystallized from methanol to give the above compound; m.p. 198-20GS .YD (, dimethylformamide).

Example 17. Greg. Butyloxycarbonyl-glidyl-gly-cyl-glycyl - alanylglydine hydrazide (BOC-GLy-Gly-Gly-Ala-Glv--NHNHj).

A solution of gOg (2.32 mmol) of BOC-Gly- -Gly-Gly Ala-Gly-OMe 1 ml (23.2 lbol) and hydrazine hydrate in methanol (30 ml) was stirred at 0 ° C for 4 hours. Evaporated. The residue is triturated with diethyl.m. ether, dried, and the above compound is obtained t. Pl. 221-223 ° C.

Example 18. Hydrazide N, 8-ditrite, ilcystenyl - N-tert.butyloxycarbonyl - lysyl-asparaginyl - phenylalanyl - phenylalanine (Trit-Cys (Tril) -Lys (BOC) -Asn-Phe-Phe-ХНХНг).

A solution of 1.25 g (1.0 mmol) of Trit-Cys - (Trit) -Lys (BOC) - Asn-Phe-Phe-OMe and 0.97 ml (20 ppm) of hydrazine hydrate in methanol (30 ml is stirred BAT 0 ° C 2 DAYS The solvent is distilled off and the residue is crystallized from ethanol / isopropyl ether to give the above compound; NMR (DM M CO de): b 1.38 (S, 9H), 7.19-7.30 (t40N).

Example 19. NS-ditrityl-disteiBilN- -greg. Butyloxycarbonyl-lysyl - asparaginyl - phenylalan.an-phen lanolil-tr, and ptofil-X -greg.butyloxycarbonyl - lysyl-Otrvg. Butyl-threonyl - phenylalanyl-O - third. Butyl-threonpl-O-Greg. Butyl - Seril-S - Tritnl-cysteine (Trit-Cys (Trit) -Lys (BOC) - -Asn-Phe-Phe-Thr-Lys (BOC) -Thr (BuO-Phe- Thr (Bu) -Ser (Bu) -Cy (Trit) OH) 0.09 -IL (0.74 mmol) Tert-butyl nitrite is added to a solution cooled to -20 ° C containing 0.2 g (0.493 mmol} Trit-Cys ( Trit) - Lys (BOC) -Asn-Phe-Phe-NHNHs in dimethylformamide (10 ml) and a 2.6N solution of hydrogen chloride in ethyl acetate (0.475 ml, 1.23. M.yul). Mix 15 MtH at -20 ° C, add 0.7. (0.493 mmol) solution H - Trr - Lys (BOC) -The (Bir -) - Phe - Thr (BuJ- -Ser (Bu) -Cys (Trit) -OH.HCOOH n 0.3 ml (1.65 .1 /. "Ol.h) of dilzol. Propylethylamine in dimethylforma mid (10 ml). The solution is stirred at -20 ° C for 1 hour and at 25 ° C for 24 hours and The residue is triturated with water, diethyl ether, cold 1N citric acid, and dried to give the above compound. NMR (CDCb); b 1.08 and 1.13 (S, 27H), 1.37 (S, 18H), 7.28 (m, BON); amino acid analysis; Lys 2.23; disteic acid 1,10; Asp 1.00; Thr 2.12; Ser 1.02; Cys 0.64, Phe 3.18.

lip 11 M ep 20. Formate S-trityl-cysteinyl-N-greg. Butyloxy-carbonyl - lysyl-asparagine d-fe1 ylal .-: 1l - phenylalanyl-tryptophyl-X -reg. butyloxycarbonyl - lysyl0-treg. butyl-threonyl - enanilil-O - tert.butnl-threonyl-0-tert. butyl - seril - S-trityl-cpstein (H-Cys (Trit) -Lys (BOC) - Asn-Phe-Phe-trp-Lys (BOC) -Thr- (Bu) -Phe-Thr (Bu) -Ser (Bu) -Cys- (Trit) -OH. HCOOH).

A solution of 0.50 g (0.192 ppm) Trit-Cys- (Trit) -Lys (BOC) -Asn-Phe-Phe-Trp-Lys (BOC) -Thr (Bu) -Phe-Thr (Bu) -Ser ( Bu), - Cys (Trit) -OH 6 ml of acetic acid / formic acid / water (7: 1: 2) are stirred for 25 ° C for 6 hours. The solvent was distilled off and the residue was triturated with diethyl ether, the above was prepared: No. Compound; A1minokiotny analysis: Lys 2.23; cysteine cyloid 1.38; Asp 1.00; Thr 2.14; Ser 0.86; Phe 3.24. Example 21. Greg. Butyloxycarbonylgly; P: I, L-gl. Icyl - glidyl - alanyl - gl.cyl-S trityl-cysteinyl-N - - / reg. Butyloxycarbonyl-isyl - asparaginyl-phenylalanyl - phenylalanyl - tryptophan-M-greg . Butyloxycarbonyl-lysyl-O-Greg. Butyl - Threonyl-phenylalanil-O-Cr0g. Butyl - Threonyl-O-tert.butyl-Seril-S - Trityl-cysteine (BOC) - -Gly-Gly Gly-Ala-Gly - Cys (Trit) - -Lys (BOG) -Asn-Phe - Phe-Trp-Lys (BOG) -Thr (Bu) -Phe-Thr (Bu) -Ser (BuO-Cys (Trtt) i-MOH). 0.04 ML (0.34 mmol) Greg. Butyl nitrite is added to the coolant, g, d н n: but to a 20 ° G solution of 0.076 g (0.176 ppl) of BOC-Gly-Gly-iGIy-Ala- Gly-NHNHj in a mixture of dimethyl sulfoxide (1 lgl), dimethylformamide (2 ml) and 2.5 n. chloride solution

i - (; iy-G-lij-Gly-Aia - Gttj-Cys-Lus - A5n-Plie-Phe-Trp-.Lys-Thr-he-Thr-8r --- Cy, 5-O.H

1t of hydrogen in ethyl acetate (0.176 ml. 0.44 mmol.) After stirring at -20 ° G for 15 minutes, a solution of 0.385 g (0.16 mmol) of HCys (Trit) - Lys (BOG) -Asn-Phe is added to the mixture. -Phe-Trp-Lys (BOG) -Thr (Bu) -Phe-Thr (Bu) -Ser (Bu) -Gys (Trit) -OH. HGOOH and 0.12 ml of diisopropylethylamine in dimethylformamide (5 ml). The solution is stirred at -20 ° G for 1 hour, then the temperature is raised to 0 ° G and stirred for 1 hour, then stirred for another 20 hours at 20 ° G. The solvent is evaporated and the residue is added to diethyl ether (100. "l). The precipitate is collected, washed water, methanol, and dried, get the above compound; amino acid Analysis: Lys 2.30; Cysteine K Slot 1.42; Asp 1.00; Thr 2.72; Ser 1.00; Giy 3.84; Ala 1.00; Phe 3.34. Note e, p 22. Cyclic disulfide GLIDYL-GLI. CYL-GLYCIL-alanyl-glycyl-cysteinyl-lysyl-asparaginyl-phenylalanyl-phenyl-aryl-tryptophyl-lysyl-threonyl-phenyl-aryl-threonyl-ser, or yl-cysteine

A solution of 0.285 g (0.103 lshoug)) BOG- -Gly-Gly-Gly-Ala-Gly-Gys (Trit) -Lys- (BOG) -Asn-Phe-Phe-Trp-Lys- (BOG) -Thr (Bu) -Phe - Thr (Bu) - -Ser (Bu) -Gys (Trit) -OH in acetic acid (200 ml) was added to a solution of 0.5% iodine in metaiol (H2 ml, 1.03 mmol) over 60 min, continue to stir- uU | -AtU-Gvy-Cys-Lys (BOC) -A3n-Phehei, -Thr (Eu) (Bu) -SBr (bu) -Su5 The solution of this cyclic heptadecapeptide disulfide (0.1 mmol ) in conc. hydrochloric acid (10 ml) was stirred at 0 ° G for 10 min. in nitrogen atmosphere. 100 ml of acetic acid and lyophilis chot are added. After re-lyophilization from water (100 ml), the residue is chromatographed on a column with chemically modified crosslinked dextran (Gefadex G-25, 3X50 cm) suspended in the lower phase of butanol / acetic acid / water (4: 1: 5). For desorption, the upper phase is used; an almost pure heptadekapeptide is obtained. The pure fractions are combined, evaporated and lyophilized to give the above compound as an acetic acid salt.

UV spectrum, A „, ah (MeOH) 283 microns (g 6702), 289 microns (e 6350). Repeated lyophilization

Sewing 60 minutes at room temperature. The solution is cooled to 0 ° G and 1 N is added. 2.06 mmol sodium hyposulfite solution, for de-lightening. The solvent was distilled off, and the oily residue was added to water (100 ml). The precipitation is filtered off and dried, and a heptadecapepeptide cyclic disulfide of the formula

the product from water gives the above compound as a free base; amino acid analysis: Lys 2,0; cysteic acid 1.42; Asp 1.09; Thr 1.89; Ser 0.91; Gly 3.64, Ala 0.98; Phe 2.95.

Similarly, you can get the above compound applied instead of iodine Rodan by the method of Hiski and GMHTa.

Example 23. N, S-dntrityl-cytosine N -treg. Butyloxycarbonyl-lysyl-asparagipyl - phenylalanyl-phenylalanyl - tryptophyl-№ -t-butyloxycarbonyl - lysyl-Otret. Butyl: yl - threonyl - phenylalanyl - ogreg. butyl - threonyl-O-greg. butyl - seryl2-tertylthioethylamide, (Trit) -Gys (Trit) -Lvs (BOG) -Asn-Phe-Phe - Trp-Lys (BOG) -Thr (Bu) -Phe-Thr (Bu) -Ser (BuO- NHGHoGH S-Tritj. Trp-Lys BSOS) Oh

13

A solution of 0.80 g (0.637 mmol) of the reptile of the pentapeptide Trit-Cys (Trit) -Lys (BOC) .-- Asn-Phe-Phe-NHNHj in Ashram (Ilfor: mother, Id (9 ml) and cooled to -20 ° C. Add to it a 2.4N solution of h.: Hydrogen sulphate in ethyl acetate (0.691 mmol), then 0.0872 ml (0.764 mmol) of greg.b of tilnitrite. The mixture is stirred for 15 min. Dari -15 ° C. A solution of 0.852 g (0.637 mmol) of H-Tr-Lys (BOC) -Thr (BuO- - Phe - Thr (Bu) - Ser (Bu) - - NHCHaCHjS-Trit in dimethylformamide (8 ml), equivalent to 0.272 ml ( 1.59 mmol) of K-ethyl-diisoprolylamine, cooled to -15 ° C, 1I is added dropwise to the reaction mixture. The stirring is continued at -15 ° C for 1 hour and at a temperature of about 15 ° C. The reaction mixture is evaporated in a vacuum, the residue is triturated with water, filtered, washed with water, dried iad (P-phosphoric acid. The residue is chromatographed with silica gel (163 g), the product is eluted with chloroform, containing 3 % (Methanol and 0.3% Liridine. The pure product is crystallized from ombsi methanol with ethanolyl ether | And get the above connection | Inen1ie with so pl. 163-180 ° C (-p-azl.).

Found,%: C 69,24; H 7.09; N 8.9.

Ci HieoNi OlgSs.

Calculated,%: С -69,85; H 7.04; N 8.76.

Pr e. M e ip 24. Forlniat 5-trityl c; istenil - S – greg. Butyloxycarbonyl - lysyl asparaginyl – phenylalanil - phenylalanyl tryptophyl –No – goth. Butnloxycarbonyl –lyl-O-greg. Butyl-threonyl - f. .butyl-threonyl-O-greg.butyl - seril - 2tr (ittilt1ioe111LILAMID: a (H - Cys (Trit) - -Lys (BOS) -Asn - Phe-Phe - Trp- -Lys (BOC) -Thr (Bu) -Pbe-Thr (Bu) - -Ser (Bu) -NHOH CH2S-Trit HCOOH).

, A solution of 0.909 g (0, i355 mmol) of underite Trit - Cys (Trit) - Cys (BOC; - -Asn - Phe-Phe - Trp - Lys (BOC) - -Thr (Bu) -Phe - Thr (Bu) - Ser (Bu) - -NHCH2CH2S-Trit IB Omeoy 20 ml of acetic acid (ants; K1ISL (Oty) water. (7: 1: 2) pvrv11esh), and incubated overnight at 1-1-2 temperatury. The solvent is distilled off, the residue triturated with (water, the precipitated K is obtained by filtration, bruised

(H leu-Gty-Gly-A; a-SVij - C. s-LH3 - A5n-- he-rhe Trp-Lus-Thr-Pns-Thr-reg-ZhSN.HK .., S)

A solution of 0.230 g (0.083 mmol)

BOC-Leu-Gly-GV4-Ala-C4y-CysiTrit) -L-Lis (BGC-Asn-Phe-Ph.e-Trp-lys (30C-Thr lBu) -Phe-Thr (Bu -) - SerCBu) - l IiCH2CH2S-Trit

AND

water 1and dried "hell p ouzyvOFOSFOR, get the above compound; amino acid analysis: Lys 2.03; Asp 1.00; Ser 0.87; Phe 2.97: cystei / nova acid 0.90; Thr 1.85.

EXAMPLE .m EXAMPLE 25 Gret.butiloksikarbonilleytsil-glycyl-Glide-Alganov - glltsil - Stritil - UiiCTeiiH.ri-N- treg.butiloksikarboNyl-lysyl-asparalianl - fe ilalanil - phenylalanyl - triptofil-: № - greg.butiloksikarbonil - lysyl - O-tt.6utkl - threonylphenylalanne-O-t-t-butyl-threonyl-O-greg. butyl - 2 - tritylthioethylamide G (BOC) -Leu - Gly-Gly - Ala - Gly - -Cys (Trit) - Lys (BOC) - Asn - Phe- -Phe - Trp - Lys (BOC) - Thr (Bu) -

-Phe - Thr (Bu) - Ser (Bu) - -NHCHsCHsS-Trit.

To a solution of 0.066 g (0.136 mmol) of ldrasil of the pentapeptide BOC-Leu - Gly-Gly-Ala-Gly-NHNHj IB cyixoiM dimethylfor: 1mide (3), cooled to -20 ° C, add 2N. 0.175 ml of hydrochloric acid in ethyl acetate, then 0.0186 ml (0.13 mmol) treg-butyl nitrite. Mixture rimsH-15 ° C 15 min. A solution of 0.315 g (0.33 mmol) of H-Cys (Trit) -Lys (BOC) -

-Asn, - Phe-Phe - Trp-Lys (BOG) - -Thr (Bu) - Phe-Thr (Bu) -Ser (Bu} - -NHCH2CH2S-Trit in dimethylforma “de (4 ml) contain 0.082 ml (0.476 mmol) 1 1-ethyldn isopropyl; 1 MI “a, cooled to -15 ° C, stirring is continued at -15 ° C for 1 hour, and the mixture is kept at room temperature overnight. The reaction mixture is evaporated in vacuo, the residue triturated with ice. on ice 1 y. limo. Nnoy acid, filtered B washed with WATER.

The solid residue is triturated with metachol and dried with l, phosphorus thioxide, beam: south of the above connection; amino acid analysis: Ly 1.88; cysteic acid 0.84; Asp 1.00: Thr 1.94; Ser 0.97: Gly 2.78; Ala 0.89; Leu 0.89: Phe APT.

Example 26. Cyclic disulfide leucylglycyl - glycyl - ala.nyl - glycylcytecnyl-lysyl - asparagilyl - phenylalanyl-phenylalanyl-tryptophyl-lysyl-threonylphenylalanyl - threonyl-seryl - 2-thioethylamide

15

in acetic acid {100 ml}, slowly add 0.211 g (0 ,.- 3) iodine to 42 ml of methanol at room temperature with vigorous stirring and mix the mixture at room temperature with iGO M.UH. Cool slowly to 0 ° C and add 1N. sodium hyposulfite solution

BOC-Leu-ily-Gly-Ala-Giy-Cij3-Ly5 (BOC) -ASn-Phe-Ihe-Trp-Lys {BOC)

-Thr isu 1- rhs-Thr (Bu) -BegSBi b-MISNzSNY

With vigorous stirring to cyclic hexadecapeptide in concentrated hydrochloric acid (7 ml) at 0 ° C, under nitrogen atmosphere, for 10 min. 90 ml of acetic acid is added and the solution is lyophilized. The residue is washed with 2% acetic acid in water and lyophilized. Then it is purified by chromatography on a column with H1I1Le modified Sephadex G-25 M crosslinked dextran (3x50 cm), which is initially equilibrated in the butanol / acetic acid / water mixture (4: 1;: 5), then balanced in the upper phase. For desorption of almost pure hexadecapeptide, the upper phase is used.The pure fractions are combined, evaporated, lyophilized and the above compound is obtained as an acetic salt.

UV spectrum: i ,,,, v (MeOH) 290 microns (e 4290); 282 microns (& 4910), 273 ir (s 4630). Repeated lyophilization of the product gives the above compound as a free base; amino acid analysis: Lys 2,01; Asp 1.35; Ser 0.93, Ala 0.99; Phe 2.52; cysteic acid Q, 6G; Thr 1.98; Gly 3.00; Leu 0.96.

Example 27. Greg. Butyloxycarbonyl-glycyl-glycyl-glycyl-alanyl-glycyl-trityl-cysteinyl-: No. -gret. butyl - threonyl-phenylalanil-O-Greg. butyl - threonyl-0-t-butyl-seryl-2-tritylthioethylamide. (BOC- -GIy -. Gly-G y-Aia - Gly-Cys (Trit) - Lys (BOG) -Asn-Phe-Phe-Trp-Lys-

(H-Glu-C 4y-Oly-Ala-Cly-Cijs-Lys-ASrt-Ke-Ph. E-Trp-Lys-Thr-Phe-Thr-Ser-NHCHgCH S 5

A solution of 0.224 g (0.083 mmol) BOG-Gly-Gly-Gly-Ala-Gly-Gys (Trit) -Lys (BOG) -Asn-Phe-Trp-Lys (BOG) - Thr (Bu) -Phe-Thr (Bu ) -Ser (Bu) - MHG: H2GH2S-Trit in 160 ml of acetic acid is slowly added to a strongly stirred solution of 0.211 g (0.83 mmol) iodine in methanol (42 ml) at room temperature

sixteen

in water to destroy excess iodine (bleaching solution).

The solvent is distilled off almost to dryness, the residue is triturated with cold water, filtered, washed with water, dried over phosphorus pentoxide, washed with ether and dried, to obtain a cyclic disulfide of the formula

- (BOG) -Thr (Bu) -Phe - Thr (BuO - -Ser (Bu) -: NHGH2GH2S-Trit).

To a solution of 0.0608 g (0.141 mmol} hydrate BAP-G1} -Gly-Gly-α-Ala-Gly-HNHNH2 in dry dimethylformamide (#} and 2 ml of dimethyl sulfoxide, cooled to -20 ° G, add 2N). hydrochloric acid in ethyl acetate (0.176 ml, 0.352 lgjoll), then 0.019 lgl (0.196 lsh./g) treg.butylnitrite.

-15 ° G 15 min. A solution of 0.327 g (0.133 mmol} H-Gys (Trit) -Lys (BOG) -Asn-Phe- -Phe-Trp-Lys (BOG) -Thr (Bu) -The-

-Thr (Bu) - Ser (Bu) -NHGH2GH2S - -Trit in dimethylformamide (4 ml} containing 0.085 ml (0.494 mmol} of N-ethyldiisopropylamine, cooled to -15 ° G and added dropwise to the reaction mixture. Continue stirring at -15 ° G for 1 h and at room temperature overnight.The reaction mixture is evaporated under vacuum, the residue is triturated with ice-cold 1N citric acid, filtered and washed with water.The solid residue is triturated with methanol and dried over phosphorus pentoxide the above compound is obtained; amino acid analysis: Lys 2.31; cysteic acid 0.84; Asp 1.00; Thr 2.26; Ser 1.1 9; Gly 4.00; Ala 0.84; Phe 3.2.

Example 28. Cyclic disulfide glycyl-glycyl-glycyl-alanyl-glypyl-cysteinyl-phenylalanyl-phenyl-alanyl-tryptophyl-lysyl-threonyl-phenyl-alan-threonyl-seryl-2-thioethylamide

I,

temperature After the addition of iodine, stirring of the solution is continued at room temperature for 60 minutes. The solution is cooled to 0 ° G and 1N is slowly added to it. sodium hyposulfite solution

water to destroy excess iodine (prior to bleaching). The solvent is distilled off almost to dryness, the residue is triturated from cold 17

water, filtered, washed with water, dried, receive cyclic disulfite

BFB-GITJ-Gly-Clg-Ala-Gly-C ys-Li sCBOO-ASH-Phe pj e-Trp-LijStBOC-Thr (Bu l-Fhe-Thr (Bu) -SeriBu) - "HCH2CH2S This cyclic hexadequa peptide is strongly mixed with 7 ml of concentrated hydrochloric acid at 0 ° C under nitrogen for 10 minutes. 90 ml of acetic acid is added to the mixture and the solution is lyophilized. The residue is washed with 2% acetic acid in water and again lyophilized. The residue is chromatographed on a column with chemically modified crosslinked dextran (Sephadex G - 25 M), first balanced in the lower phase of the butanol / acetic acid / water system (4: 1: 5), then in the upper phase of this system. The upper phase is used to desorb almost pure hexadecapeptide. The pure fractions are combined, evaporated and lyophilized to give the above compound as an acetic acid salt. UV spectrum: Yatah (MeOH) 288 f / s (e4870), 280 microns (e5575), 274 microns (eBSvO), 268 microns (e5185), 265 microns (s4955). Repeated lipophilization of the last product from water gives the aforementioned compound, in the form of a free base; amino acid analysis: Lys 1.72; Asp 1.00; Ser 0.73; Ala 0.69; Phe 2.78: cysteic acid 0.57; Thr 1.69; Gly 3.59. Example 29. Greg. Butyloxycarbonylglycyl-glycyl-alanyl-glycyl - S - tritylcysteinel. - O - t / oeg. Butyl-threonyl-O-hot. Butyl-serial-2 - tritylthioethylamide. (BOS-

(.H-C-ly-C-ly-Ala-GXy-Cys-LiiS-A5rx-Phe-Phe-rrp-Lys-Thr-Ehe-Thr-3er-MCH, CH, S)

0.860 g (0.3 mmol BOG-Gly-Gly-Ala-Gly-Gys- (Trit) -Lys (BOG) -Asn-Phe-Phe-Trp-Lys (BOG) -Thr (Bu) - Phe-Thr ( Bu) -Ser (Bu) -NHGH2GH2S is dissolved in 150 L1L of glacial acetic acid and added dropwise with stirring, over one hour to a solution of 0.5% iodine in methanol (150% ugly, 30% lols). at room temperature. GMecb

BOt - Gly-Gly-Ata- & l.y-Ci, 3-LyalBOC1-Asu-1, e-Ph, e-: rj; -LijS (BQCi-Thr (Bu) -Phe-Thr (BuM- Sei- Su) -NliCHjCH 3

18

formulas

continue stirring for another 45 minutes, cool on ice and add 1 and. an aqueous solution of sodium hyposulfite (6 ml to destroy excess iodine (before decolorization of the solution). The solvent is distilled off, the residue is triturated with water, dried and the dry product is triturated with isopropyl ether, to obtain a cyclic pentadekapeptide disulfide of the formula Gly-Gly-Ala-Gly-Cys ( Trit) - Lys - (BOG) -Asn - Phe - Phe - Trp-Lys- (BOG) - Thr (Bu-) - Phe-Thr (Bu) - Ser (Bu) - NHGH2GH2S-Trit). To a solution of 800 mg (0.58 mmol) of BOG-Gly-Gly-Ala-Gly-Gys (Trit) -Lys- (BOG) -Asn-Phe-Phe-NHNHa in 12 ml of dimethylformamide at -20 ° G, add with stirring 1.85 n. a solution of hydrogen chloride in ethyl acetate (0.795 ml, 1,475 mmol). The temperature of the mixture is cooled to -15 ° G, 0.081 ml (0.71 mmol} of hot butyl nitrite is added, and the solution is stirred for 15 minutes. A solution of 0.52 g (0.637 mmol H - Trr - Lys - (BOG) - Thr (Bu) - Phe-Thr (Bu) - Ser- (Bu) NHCHaGHaS-Trit and 0.354 ml (2.06 mmol N-ethyldiisopropylamine in 6.0 ml dnmethylformamide is cooled to -15 ° G and added to the above reaction mixture. GMecb is stirred at - 15 ° G for an hour and at 25 ° G for 18 h. The solvent is distilled off, the residue is triturated with ice-cooled citric acid, filtered, washed with water, methanol, dried and the above compound is obtained. analysis: Lys 2.01; Asp 0.97; Thr 1.60; Ser 0.65; cysteic acid 0.87; Gly 2.92; Ala 1.00; Phe 2.97. Example 30. Cyclic disulfide glycyl-glycyl-alanyl - Glycyl-cysteinyl-lsyl-asparaginyl-phenylalanyl-phenylalanil-trintofil-lysyl-threonyl-phenylalanyl-threonyl-seryl-2-thioethylamide.

nineteen

23 ml of cold hydrochloric acid hydrochloric acid is added to this mixture, the mixture is cooled with an ice ball and under a nitrogen atmosphere, and with Pe-re) mesh, for 10 min, 300 ml of glacial acetic acid and solution lyof: ilizuyut. Residue; pa) SST (rut in water and again lofiz.izuyut. The residue is dissolved in a 0.01 iH. Aqueous solution of ammonium acetate, placed in a column with carboxymethylcellulose (Whatman GM-23, 2.5x30 cm). Pure compound evil "0.06 n. acetate and ammonium bufrom.p. The purified substance is lyophilized: from WATER AND an acetic acid salt is obtained, the above compound is in the form of white T1Head; UV spectrum: I, groin (MeOH) 282 * h (e 5120), 289 g / c (e 4610). Re-lyophilization of the product; from the water gives the above-mentioned compound in the form of a free one-jar "I. Amount of acidic analysis: Lys 2.06 ; Asp 1.02; Thr 1.75; Ser 0.91; c stennovaya acid 0.73; Gly 3.00; Ala 1.10; Phe 3.11.

At 1M1NL BiMrecTo iodine Rodanide by the method of Hismi and Smith, it is also possible to obtain the above compound.

Example 31. Methyl eflr a-cetyl (S-trityl) - cysteinyl-M - Greg. Butoxycarbanyl-lysyl - Aoparal-nyl - phenylalanylphenylalanine (Ac - Cys (Trit) - Lys - (BOS) -Asn-Phe-Phe-OMe ).

A solution of 0.191 2 (0.5 mmol) of p-nitrophenylacetate in 4 ml of dimethylfrmamide is added at 0 ° C to a solution of 0.750 g (0.698 mmol) of H - Cys (Trit) - Lys (BOC) - Asn-Phe-Phe-OMe HOAc and 0.1 ml of netilmorpholine. After being stirred at 0 ° C for 24 hours, the solvent solvent 1 was distilled off by an iB vacuum. The residue is dissolved in 3 ml of methanol, and slowly added to 200 ml of diethyl ether. The precipitate was collected and 1IZ of methanol crystallized to give the above compound with m.p. 219.5-22GS; -21.6 ° (, dimethylformamide).

The same abraem, p-n-nitrophenyl esters of murine, propioic, oily, isobutyric, trimethylacetic, kalonovoy or benzoic ester, instead of l-n; nt1-phenylacetate, receive the corresponding compounds of the above formula, where instead of ac- Fprmyl, propylene, butanoyl, zo-butanoyl, taivaloyl, hexanoyl or benzyl.

Example 32. Acetyl- (S-trityl) -cysteinyl-N - butoxycarbonyl - lIzi, l - aaparati or - phenylLan1Il-phenyl.alanine (Ac - Cys (Trit) - Lys (BFB) - - Asn-Phe-Phe- iNHNHz).

(le-C j $ -Iijs Asn-Phe-Rie-Thr-1 5-Thr -T -e-Tlt.r Se-ThOng 1

20

A solution of 0.40 g (0.379 mmol) of Ac-Cys- (Trit) - Lys (BFB) -Asn - Phe - Phe - -OII and 0.37 ml (7.58 mmol) of hydrazine hydrate in methanol (15 ml) is stirred 5 at 0 ° C 48 hours The precipitate is collected, dried and receive the above compound, with so pl. 236-237 ° C; -28.6 ° (, dimethyl form amide).

In the same way, using the appropriate starting materials, compounds are obtained in which, instead of Ac-farmil, propionyl, butanoyl, isobutanoyl, pivaloyl, hexanoyl or 1 benz, onl.

PRI me R 33. Acetyl-5-trityl-cysteinyl-H-reg. Butoxycarbonyl-lysyl-asp .. raginyl-phenylalanyl-phenyl-aryl-tryptophyl-N-guoate. Butoxycarbonyl-lysyl-Otret. -Butyl-threonyl- phenylalanil - O-tert.p butyl-Tr | eon | Il-seril-2 - 11-ethylthiostila1MID (Ac - Cys (Trit) - Lys (BOS) - Asn - -Phe-Phe-Trp-Lys (BOS) -Thr (- Phe-Thr (Bu) - Ser (Bu) - NHCHj- -CHsSTrit).

K pacTiBopy 0.240 g (0.227 mmol) Ac-

Cys (Trit) -Lys (BFB) -Asn-Phe-Phe- -iNHNHg in CyxOM dimethyl L amide forms (2 ml) and dimethyl sulfoxide (1 ml), cooled to -20 ° C, add 2N. hydrochloric acid in ethyl acetate (0.273 mmol), then 0.0312 ml (0.273 mmol) of tert-butyl nitrite. The mixture is stirred for 15 min at −15 ° C. A solution of 304 g (0.227 mmol) of H-Thr-Lys (BFB) -Thr (Bu) -Phe-Thr- (Bu) -Ser (Bu4-NHCHaCHsS-Trit in Dimethylfor) Ma mide (3 ml) containing 0.097 ml (0.568 mmol) of N-ethyl lId of nosopylamine, cooled to -15 ° C, is added to the above reaction mixture. The mixture is stirred at -15 ° C

1 hour and at 25 ° C for 20 hours. The solvent is distilled off in VA, Kuum, the residue is triturated with ice on ice 1. mono acid, filtered, washed with water | and dried over the phosphorus. The solid residue is triturated with α-methanol, filtered and dried over phosphorus pentoxide to give the above compound; amy: no acid analysis: Lys 1.82; Asp 1.00; Ser 0.76; cysteiH101va acid 0.93; Thr 1.87; Phe 3,10.

In the same way, by using the appropriate starting materials, compounds of the formula P are obtained, where imesto Ac-formyl, propionyl, (butanayl, isobutanoyl, pivaloyl, hexachhoyl or benzoyl.

Example 34. Cyclic disulfide acetyl-cysteinyl - lysyl-asparaginyl - phenylalanyl - phenyl-anil - tryptophyl-lysyl - threonyl-phenylalanyl - threonyl-seryl2-thioethylamide;

21

A solution of 0.250 g (0.110 mmol) Ac-Cys- (Trit) -Lys (BOG) -Asn - Phe-Phe - -Trp - Lys (BOG) -Thr (Bu) - Phe-Thr (Bu) -Ser (Bu) - NHGHjGHaS-Trit in acetic acid (61 ml), mixed; a, under stirring (0.278 g), 1.1 mmol of iodine in Methanol (56 ml) was added at 25 ° G. iOo partly the added solution kept stirring 25 ° G to teAt-Cy5-Lyf; (BOC) -A3n- he Phe-Trp-Ly3tbOC -Thrl.Ba) -rhe-Thr Bu) - $ ep (Bu), CHoS

To the cyclic peptide, 9.1 ml, pIr of HVAC hydrochloric acid, p-ri 0 ° G, under nitrogen atmosphere for 10 minutes, are added. With vigorous stirring, 119 ml of uxuanium acid are added and the solution is immediately lyophilized. The residue is dissolved in water and reappointed. The residue is then dissolved in IB of the upper phase of the solvent mixture of the buta, nol / acetic acid / iodine a (4: 1: 5) and placed in Olon.ku with chemically modified Cipher dextrus (Gefadex G-25 M), prepared with in the form of a suspension of the lower phase of the umeo solvent. The upper solvent phase is used to desorb the undecapeptide. The fractions containing pure product are combined and evaporated in vacuo. The residue was triturated with diethyl ether, dissolved in 5% acetic acid, lyophilized, and the above compound was obtained as the acetic acid salt;

UV spectrum: max (MeOH) 290 microns (e4990), 282 microns (e 5455), 274 microns (e 5095).

Preparation in the form of the acetic acid salt is repeatedly lyophilized from water and the above compound is obtained in a free-ice form; amino acid analysis: Lys 1.93; Asp 1.00; Ser 0.84; cysteic acid 0.80; Thr 1.84; Phe 2.97.

In the same way, using other starting materials, the corresponding compounds of the formulas III and L are obtained, where R is H and R is NHR 2 where formyl, propionyl, butanoyl, isobutanoyl, plvoaloyl, hexanoyl or benzoyl.

Approx. 35. Pentachlorophenyl. Ethyl ester of Z-tritylthioIropionic acid (Trit-SHsGHaGOORhGIs).

To a solution of 1.0 g (2.87) of 3-tritylthiopropionic acid in dry tetrahydrofuran (25 ml). Add 0.765 g (2.87 mmol) of pentachlorofelol. G.Mecb is cooled to 0 ° G, 0.596 g (2.87 ljol) of dihydrocyclohexylcarbodiimide are added to it and stirred for 1 hour at 0 ° G, then for 1 hour at 25 ° G. Then the mixture is cooled to 0 ° G , f; iltrate, the filtrate is evaporated in vacuo. The residue is crystallized from ethyl acetate and get the above connection with so on. Il. 154-156 ° G.

22

chevy hour The solution is cooled to 0 ° G; slowly add bH. a solution of sodium hyposulphite in water to destroy excess iodine (decolouration of solution). The solution was distilled off in vacuo, the residue (triturated with iC water, filtered, washed with water, dried over phosphorus pentoxide, was prepared (Cyclic Lock for 1-1 decapeptide of formula

Ex. 36. Methyl 3-tritylthiopropponyl-N-ester - greg. Butoxycarbonyl lysyl-asparapinyl - phenylalayl - .phenylalanine. (Trit-SGH2GH2GO-Lys (BOG) -Asn-Phe-Phe-OMe).

A solution of 0.579 g (1 mmol) of penta.chlorophenyl ester 3-tritylthiopropionic acid, 1 mmol H-Lys (BOG) -Asn-Phe- -Phe-O. Me-OH Ac, and 0.14 (1 mmol) triethylamine. stir at 25 ° G 3

DAY. The solvent is distilled; in vacuum, the residue is triturated with chilled ice. water solution of lemon acid, filtered, pramy.yvayut BO.ODY and dried, caustic potassium. The crystalline solid is made from methanol to give the foregoing compound 1BNI with m.p. 215-220 ° G.

Example 37. G. hydr. Azid 3-tritylthiopropion - N - greg.butokspkarboiil - lysyl-asparagaiyl - phenyl.lanil - phenylalanine (Trit - SGHsGHaGO - Lys (BOG) - -Asn-Phe-Phe-XHNHa).

A solution of 0.900 g (0.9 mmol) of Trit - -SGHsCHsGO-Lys (BOG) -Asn - Phe- -Phe-OMe and 1 ml of hydrazine dihydrogenate in dimet. Forma; m "de (20 ml) is stirred by sp. And 25 ° C 20 hours The solvent is distilled off in vacuo. The residue is triturated with cold water, filtered, washed with water. And dried,. The above compound with t is obtained.

ll 225-235 ° G.

Example 38. 3 - tritylthiopropioNILN -greg.butoxycarbonyl-lysyl-asparaginyl-phenylalanyl-phenylalanyl-tryptophyl-N -butoxpcarbon41l-lysyl-O-tert-butyl-treonpl-phenyl-alapyl-O-gret.butyltro-redyl-redyl-redyl-redyl. - tritylthioetplamnd (Trit-SGHaGHoGO-Lys (BOG) - Asn-Phe - Phe-Trp - Lys (BOG) - -Thr (Bu) - Phe-Thr (Bu) - Ser

(Bu) -NHGHaGHaS-Trit).

To a solution of 0.5 g {0, мм mmol) Trit-SGHaGH.GO-4ys (BOG) - Asn - Phe - -Phe-NHNHj, in 20 ml of dimethylformamide and 5 ml of dimethyl sulfoxide, o.lal;

to -20 ° G, a solution of 1.4 N is added. 0.895 ml of hydrochloric acid B in ethyl acetate, then 0.069 ml (0.6 mmol) of grvg.butylnitrite. GiMecb is stirred at -15 ° G for 15 min and rast is added cooled to -15 ° G

Thief 0.67 g (0.5 sylux) H-Tgr-Lys (BFB) - -Thr (Bu) -Phe-Thr (BuO-Ser (BuO - -NHCH2CH2S - Trit "0.214 ml (1.25 mmol) N -et.ild: and, isopliplIla; MiEHa In 10 ml of dimethylformamide. The reaction mixture is neipeMie sewn at -15 ° C for 1 hour at 25 ° C for 20 hours, then evaporated to a vacuum. The residue is triturated with ice-cooled aqueous solution 1 cooled on ice. L. m., IMNIs, is fluttered, washed with water, “dried” by the adhesion of the thiomysyu% PSCHSN 00-Vr-ASn-The-Fne-trp-L -jg-Thr-TheI.

A solution of 0.5 g (0.216 mtol) Trit-

-ССНгСНгСО-Lys (BOS) -Asn - Phe-

Phe-Trp-Lys (BOG) -Thr (Bu) -Phe - Thr (Bu) -Ser (Bu) -NHCHzCHjS-Trit

in 100 ml of acetic acid slowly add 15

Pour to a stirred solution 0.547 g

(2.16 mmol) iodine in PO ml of methanol at

25 ° C. At the end of the addition solution

continue to stir at 25 ° C for 1 hour. & CH2CH ,, CO-Lys (BOC) -Asn-Phe - Thr (Bi) - Sei- (Bu) The decapeptic mixture is stirred in ml of concentrated hydrochloric acid at 0 ° C under nitrogen atmosphere for 10 minutes, 200 ml of acetic acid and the solution immediately lyophilized. The residue is dissolved in water and lyophilized, then the residue is dissolved in the upper phase of the solvent mixture butanol acetic acid / water (4: 1: 5) and placed in a column with chemically modified crosslinked dextran (Sephadex C-25 M) prepared in the lower phase of the mixture solvents. The upper phase of this mixture is used to desorb the decapeptide. The fractions containing the pure product are combined and evaporated in vacuo. The residue was dissolved in 5% acetic acid and lyophilized to give the above compound as an acetic acid salt. UV spectrum: Xtah (MeOH) 290 microns (s4920), 282 microns, (e 5390). This compound, in the form of an acetic acid salt, is repeatedly lyophilized from water and the above compound is obtained as a free base; amino acid analysis: Lys 1.97; Asp 1.00, Thr 1.64; Ser 0.65; Phe 2.94. Example 40. Methyl ether a, a-dimethyl-3,5-dimethoxybenzyloxycarbonyltryptophyl-N-gret. Butyloxycarbonyl-lysyl-O-treg. Butyl-threonyl-phenylalanyl-tert-butyl-threonyl-O-gret. Butyl-seryl S-trityl cysteine. (DDCbo - - (BOC) -Thr (BuO-Phe - Thr (Bi) - -Ser- (Bu) -Cys (Trit) -OMe).

24

phosphorus. The solid residue is triturated with cold metal and Dried, and the above compound is obtained; amino acid analysis: Lys 1.99; Asp I, il5; Thr 1.73- Ser

0.67; Phe 3.00.

P p And M e p 39. Cyclic disulfide of Z-chioprolnoyl-lysyl-asparaganyl - phenylalanil - phenylalanil - tryptophyl - lysyltrbonyl - phenylalanyl - threonyl - ser1IL-2-ethyl ethyl amide;

The solution is cooled to 0 ° C and 1N is slowly added. an aqueous solution of sodium hyposulfite to destroy excess iodine (until the solution becomes colorless). The solvent is distilled off in vacuo and the residue is triturated with water. The precipitate is filtered off, washed several times with water and dried over phosphorus pentoxide, to obtain a cyclic protected decapeptide of the formula Thr-Ser-NHCH OSH) I-Trp-Lys BOS) -Hrb GKeCH CH A solution of diazomethane in ether is added to a solution of 12.6 g ( 0.795 mmol DDCbo - Thr-Lys (BOC) -Thr (Bu) - -Phe-Thr (Bu) -Ser (Bu) -Cys (Trit) - -on in 10 ml of methanol at 0 ° C. The mixture is stirred for 1 hour and The residue is chromatographed on silica gel using 3% methanol and 0.5% pyridine in chloroform for elution. The eluate is evaporated to give the above compound, NMR (CDCIs): 61.13 (S, 18H), 1.27 (S, 9H ), 1.48 (S, 9n), 1.77 (S, 6H), 3.68 (S, 3H ), 3.79 (S, 6H), 7.1-7.6 (m, 23H). In the same way, instead of diazomethane, diazoethane, 1-diazopropane, 2 diazopropane, 1-diazobutane, 1-diazoisobutane, 1-diazopentane are used instead , 4-diazo-2-methyl butane, 1-diazo-hexane, 1-diazo-heptane or 1-diazo-diazo-hexane, 1-diazo-heptane or 1-diazo-octane, the corresponding ethyl octane is obtained, the corresponding ethyl, propyl, isopropyl, - are obtained butyl, isobutyl, propyl, isopropyl, amyl, 2-methylbutyl, hexyl, heptyl, and RiTWIT-V TliTOTTJT II GTTT TT TatTTTTT octyl esters of the indicated compound. Example 41. S-tritylcysteine decyl ether (H - C3S (Trit) - OCH2 - (CH2) 8CH3). A solution of 0.894 g (3 mmol) cysteine decyl ester hydrochloride 0.78 g (3 mmol} triphenylcarbinol and 0.42 ml of a mixture of boron trifluoride in acetic acid (5.2 ml) is stirred at room temperature for 1 hour. The solvent is from

vacuum and the residue is chromatographed on silica gel, using a 40% ethyl acetate / hexane solution containing 0.1% triethylamine for elution. After evaporation of the eluate, the above compound a 1.5 +25.84 (, CHCis) NMR (CDCis) 60.89 (t, I 5HZ, 3N), 1.3 (S, 16H), 1.55 ( m, 2H), 2.55 (m, 2H), 3.25 (2d, I 5HZ, IH), 4.1 {t ,, 2H).

In the same way, but replacing the starting material with an equivalent amount of cstein tetradecyl ether hydrochloride, tetradecyl ether is obtained.

tritylcysteine, "+ 13.8 ° (, СНС1г). NMR (CDCls) 60.89 (t, I 5HZ, ЗН), 1.3 (S, 24H), 1.56 (m, 2H), 2.55 (m, 2H), 3.23 (2d, I 5HZ, IH), 4.1 (tI 6HZ, 2H).

In the same manner, the corresponding nonyl, undecyl, dodecyl, and S-tritylcysteine tridecyl esters are prepared.

The combination of the above esters of S-tritylcysteine with a hexazptidide hydrazide of the formula DDCbo-Trp-Lys (BOC) - Thr (Bu) -Phe-Thr (Bu) -Ser (Bu) - NHNH2 gives the azide method nonyl, decyl, undecyl-, dodecyl, tridecyl and tetradecyl esters of the corresponding compounds.

Example 42. Tryptophil-NO. -Greg.-butyloxycarbonyl-isyl-0-warm methyl formate. butyl - threonyl-phenylalanil-0-t. butyl threonyl-O-greg. butyl serine S-trityl cysteine; (H-Trr-Lys (BOC) - Thr (Bu) -Phe-Thr (Bu) -Ser (Bu) - Cys (Trit) -OMe - HCOOH).

A solution of 1.11 g (0.685 lshol) DDCbo-

-Tgr - Lys (BOS) - Thr (Bu ") - Phe-

-Thr (Bu) -Ser (Bu) -Cys (Trit) -OMe in 10 liters of formic acid / acetic acid / water mixture (1: 7: 2) is stirred at room temperature overnight. The mixture is evaporated and the residue is triturated with water. The solid is filtered and dried in vacuo to give the title compound.

UV spectrum;). Shah: MeOH: 290 g: (eBZB), 280 l4 / s (e 8470), 273 microns (at 3720), 216 microns (e 78400).

In the same way, using the other heptapeptides of Examples 40 and 41 as starting materials, ethyl, propyl, isopropyl, butyl, isobutyl, amyl, 2-methylbutyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl, tridecyl and tetradecyl ethers of these compounds.

Example 43. Methyl ether treg.butyloxycarbonyl-alanyl-glycyl-S-trityl-cysteinyl- - ° -tert, butyloxycarbonylisyl-asparaginyl-phenylalanyl-phenylalanyl-tryptophyl-N-tert, butyloxycarbonyl-lysyl-0-tert. -O-treg.butyl-seril-S - tritylcysteine (BOS -

26

 - Cvs (Trit) -Lys (BOC) -Asn- -Phe-Phe-Trp-Lys (BOC) -Thr (Bu) - Ser (Bu) -Cys (Trit) -OMe).

To a solution of 0.775 g (0.624 g. Chol) of heptayetiide BOC-Ala-Cys (Trit) -Lys (BOC) Asn-Phe-Phe-NHNH2 heptazide in 9 ml of dpmethylformamide, cooled to -20 ° C, is added 2.1 n anhydrous hydrogen chloride in ethyl acetate

(1.56 mmol), then 0.0863 ml (0.749 mmol), treg.butylnitrite, the mixture is shifted at -15 ° C for 15 minutes A solution of 0.900 g (0.624 mmol) of heptapepyl methyl ester H-Thr - Lys (BOC) -Thr (Bu-) -

Phe - Thr (Bu) (Bu) -Cys (Trit) - -OMe. HCOOH and 0.373 ml (2.184 mmol) of N-ethyldiisopropylamine in 5 ml of dimethylformamide are cooled to -15 ° C and added to the above solution at

-15 ° C.

The resulting mixture was stirred at -15 ° C for 1 hour and at room temperature for 3 days. The solvent is distilled off in vacuo, the residue is triturated with 1N. citric acid, filtered, the precipitate washed with water, and dissolved over phosphorus pentoxide in vacuum. The dried residue is triturated with a small amount of methanol, filtered and dried, to obtain the above compound, amino acid analysis;

Lys 2.06, cysteic acid 1.75, Asp 0.94, Thr 1.93, Ser 0.97, Gly 1.00, Ala 0.88, Phe 3.10.

In the same way, using other heptapeptide alkyl ethers, ethyl, propyl, isopropyl, butyl, isobutyl, amyl, 2-methylbutyl, hexyl, heptyl, OKTIL-, nonyl, decyl-, undecyl, dodecyl, tridecyl and tetradecyl esters of these compounds.

Example 44. Methyl ester of cyclic disulfide tert, butyloxycarbonyl-alanyl-glycyl-cysteinyl-N-tert, butyloxycarbonyl-lysyl-asparaginyl-phenylalanyl-phenylalanyl-tryptophyl-N-tert.butyloxycarbonyl-lysyl-O-gre butyl-threonyl-phenylalanil-O-tert. butyl-threonyl-ogret. butyl-seryl - cysteine (BOC-Ala- -Gly-Cys-Lys (BOC) -Asn-Phe-Phe-

- Trp - Lys (BOC) -Thr (Bu) -Phe-Thr (Bu) -Ser (Bu) -Cys-OMe).

0.898 g (0.345 mmol) of the BOC-Ala-Gly-Cys (Trit) -Lys (BOC) -Asn-Phe-Phe-Trp- tetradecapeptide methyl ester

-Lys (BOC) -Thr (Bu) -Phe-Thr (Bu) -Ser (Bu) -C3s (Trit) -OMe is dissolved in 200 ml of warm acetic acid. The solution is cooled to room temperature and added dropwise to a solution of 5% iodine in methanol (175 mg, 3.45 mmol) over 1 /. The mixture was stirred for another 1 h, cooled to 0 ° C and 1 n was added. 15 ml of an aqueous solution of sodium hyposulfite to destroy excess

iodine. The solvent is distilled off, the residue is

rubbed with water and dried over phosphorus pentoxide in vacuo, the above compound was obtained.

Thus, using other linear zinc, tetradecapeptide esters, ethyl, propyl, isopropyl, butyl, isobutyl, amyl, 2-methylbutyl, three (H - AlQ - Giu-CLiS-LyS. -Asn-The-i-lie-Trp-Lyg-Thr - The-Ti4r-S r-Sch-OL-1

To a solution of 0.345 mmol cyclic oxygen:; f BCG-Alc-Glg-Cg. tEOij) -Asn he- Iie-TrpZeg Ei) -Cif $ -Ohm (0.345 mmol) in 29 ml of concentrated hydrochloric acid under stirring under nitrogen atmosphere for 10 min at 0 ° C, glacial acetic acid (290 ml) and the solution is lyophilized. The residue is dissolved in 5% acetic acid and lyophilized again. The residue is redissolved in a 0.2 M solution of ammonium acetate, added to a carboxymethyl cellulose column (Whatman CM-23) and eluted with 0.2 M ammonium acetate buffer. The eluate is lyophilized, the residue is subjected to separation chromatography on a column with chemically modified ex-dextran (Sephadex G-25M), initially balanced with the lower phase of butanol / acetic acid / water mixture (4: 1: 5), then to desorb the somatostatin methyl ester using the upper phase . Pure fractions are combined, bulked and lyophilized to give the above compound as an acetic acid salt; UV spectrum: Xshah (MeOH): 290 microns (s 4345), 287 microns (& 4385), 268 microns (e 4090). 265 microns (e 3900), 215 microns (e 38305). NMR (DMSO-dG) b 3.63 (S, 3N, OCH3). Repeated lyophilization of this salt from water gives the above compound as a free base. Amino acid analysis: Lys 2.00; csteinopic acid 1.28, Asp 0.95, Thrl, 91, Ser 0.97, Gly 1.00, Ala 0.94, half Cys 0.30, Phe 3.01. In the same way, using other cyclic disulfides of formula III, ethyl-, propyl-, isopropyl- is also obtained. butyl, isobutyl, amyl, 2-methylbutyl, hexyl, heptyl, octyl, nonyl, decyl, unSCH-CH (h) CO-Lys - A3u-Phe-Ph, e-Trp lyS -Thr Pb.e-rht-Ser-MCH (B.} CH, S

li 1

28

decyl and tetradecyl ethers corresponding to the above compound.

Example 45. Somatostatin methyl ester; alanyl-glycyl-cysteinyl-lysyl-asparaginyl-feiyl-alanyl-phenyl-alayl-trnpto-phyl-lysyl-threonyl-phenyl-alyl-threonyl methyl serine-cysteine methyl ester methyl ester methyl ester;

Claims (1)

1 Lys SOO} -TIiri-Sa) - he-Tlii-, decyl, dodecyl, tridecyl and tetradecyl esters, corresponding to the above .viy compound. Example 46. Somatostatin methyl ester; methyl ester of the reduced form of alanyl-glycyl-cysthenyl-lysyl asnaraganyl-phenylalanyl-phenyl-aryltrintophyl-lysyl-threonyl-phenyl-alyl -treoyl-seryl-cnstein (H-Ala-Gly- -Cvs-Lys-Asn-Phe-Phe-phy-stean-H-Ala-Gly- -Cvs-Lys-Asn-Phe-P-peptyline (H-Ala-Gly-Cvs-Lys-Asn-Phe-P-peptyline) Phe-Thr-Ser-Cys-OMe). According to the procedure of Example 45, but replacing the cyclic disulfide with an equimolar co. KiKCCTiiOM of the compound BOC-Ala-Gly- -Cys (Trit) -Lys (BOC) -Asn - Phe- -Phe - Trp-Lys (BOC) -Thr (Bu) - -pjje - Thr (Bu) -Ser (BuO-Cys - the aforementioned coefficients are obtained by treating the somatostatium methyl ester from Example 45 with dithiothreitol or sodium bisulfite with the hydrolysis of the dithiosulfate derivative obtained in this way ;, see the above compound. In the same way, but using other somatostatin alkali ethers of the formula I, where R is IT-Ala-Gly-NH, B is COOAik, obtained in Example 45, or other linear sewn tetras Adecapeptide esters, ethyl, propyl, isopropyl, butyl, isobutyl, amyl, 2 methyl, ochyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl, tridecyly tetradecyl esters, corresponding to the above compound. Formula of the invention 29 where Ri is H-Gly-Gly-Ala-Gly-NH, li-Gly-Gly-Gly-Ala-Gly-NH or H-Leu-Gly-Gly-Ala-Gly-NH; W is H or COOH; or R is H or NHR3, where R is acyl or benzoyl; R is hydrogen; or R1 is H-Ala-Gly-NH; R2 is COOAik, where Alk is a chain-chain or branched Ci-Cu alkyl, characterized in that the linear peptide of the formula Trt is SCH2CH (R4) CO-Lys (BOC) -Asn-Phe-Phe-Trp - Lys (BOG) -Thr - ZSNSN (D) CO-Lys (BOG) -A3r.-Phe-Phe-Trp-L 4S (BOC) -Thr (Ba) -Phe-Thr (Bi) (Zi) YSTERE11 (W) where R2 and R are as defined above, followed by removal of all remaining protecting groups with hydrochloric acid. 04.06.75 with R1-H-Gly-Gly-Ala-Gly-NH, H-Gly-Gly-Gly-Ala-Gly- thirty (Bu) - Phe - Thr (Bu) - Ser (Bu) - NHCH {R2) -CHaS-Trt (II), where R2 is H or GOOH; R is BOG-Gly-Gly-Ala-Gly-NH; BOG-Gly-Gly-Gly-Ala-Gly-NH or BOG-Leu-Gly-Gly-Ala-Glv-NH; or R2- H; R is H or NHR, where Rs is as defined above; or R2 - GOOAlk; R - BOG-Ala- -Gh-NH; oxidized with iodine or rhodanide to obtain the corresponding cyclic disulfide and derivative of the formula -NH or H-Leu-Gly-Gly-Ala-Gly- -NH; 07/08/75 RI - hydrogen or NHR, where R3 is acyl or benzoyl, R2 is hydrogen; or R1 is H-Ala-Gly-NH; R - GOOAlk, where Alk is an alkyl group with straight or branched chain Ci-gh.