SU876641A1 - Derivatives of s-pyrrolidonemethylcystein as intermediate products for synthesis of cystin-containing peptides - Google Patents

Description

ICE for the production of citrine-containing lientides.

The goal is achieved by the proposed derivatives of S-pyrrolidone methylcysteine of General formula IX-U-H-CH-CO-Y:

I

fi

$

,,

N.

Lj °

where X is ČSNz) ASOCO (first) or HC1, Y is OH or OSJs,

ABOUT

s

-Cjj -jl - pyrrole and n don methyl (Ruth),

The compounds of the indicated general formula are obtained by the known (4) methods according to the scheme

HoeNH-CH-oooH

CHa-SH

Cysteine hydrochloride. Mol, weight 157.62,

BNNS I

SNGON

N-hydroxymethylpyrrolidone

. -

NSHS - SN-Soon CIS

S-CHj- /

about

Hydrochloride S-Ruth-Cysteine Mass Mol. 254.74

soce2, weoH

NSMS-Mr.-SN-SOOSNz

CHi

rv S-CH2. .S-Ruth-Cysteine Methyl Ester Hydrochloride, Mol. 268.76 Mol.

LCH bC-OCOD O L -t-butylpyrocarbonate

(CHCO COCO -VA- CH-COOH Io

sc. „

-S-CHj-T

 ff o C - S-Ruth-cysteine AI Mol. weight 318.39.

S-Route-cysteine (II) hydrochloride is obtained by reacting cysteine hydrochloride and N-hydroxymethylpyrrolidone at a molar ratio of 1: 1.1 for 30 minutes. Yield 90%.

Synthesis of S-Ruth-cysteine (III) methyl ester hydrochloride is carried out by esterification (I) in methanol in the presence of thionyl chloride with a yield of 99.6%.

By reacting (I 1) with di-tert-butyl pyrocarbonate in the presence of sodium hydroxide for 2.5 hours, Ba-c-S-Ruth distein is obtained (IV) in 98% yield.

Checking the effectiveness of S-Ruth-cysteine derivatives in the synthesis of cysteine-containing peptides shows that the Boc-S-Pym-cysteine condensation reaction with S-Ruth-cysteine methyl ester hydrochloride by the method of mixed anhydrides (with isobutyl chloroethylene ester in the presence of N-methylmorphine) passes with a rate of 90%.

Cleavage of the S-protecting groups from the Boc-S-Ruth-cysteinyl-S-Ruth-cysteine (v) methyl ester obtained and the disulfide ring closure are performed by iodine in methane for 1.5 h with a yield of 92%.

The use of S-Pym-cysteine derivatives as starting materials for the synthesis of cysteine-containing peptides makes it possible to obtain the above-higher cyclic disulfide of Boc-cysteinyl-cysteine methyl ester (VI) with a total yield of 73%.

Thus, the invention provides a significant positive effect. The yield of cystine-containing dipeptide in comparison with the known increases by 51%, a considerable simplification of the process associated with the use of 5-pyrrapidone-methylcysteine derivatives stable under the conditions of peptide synthesis is achieved. .

The advantages of the new compounds are their good crystallizability, high yields, resistance to the effects of various reagents used in the synthesis of peptides (the action of acids, caustic alkali, triethylamine, ammonia, hydrazine hydrate) and the selectivity of cleavage of 5-protected groups with iodine in methanol.

Claims (2)

The stability of the S-Route group is due to the presence, in the molecule of a 5-membered heterocyclic ring, pyrrolidone, in which the distribution of electrons contributes to the stabilization of the molecule. S - Root-derived cysteine-white crystalline substances, soluble in alcohols, ethyl acetate, dimethylformamide, water. Insoluble in ether, petroleum ether, hexane. The structure of new compounds was confirmed by IR, PMR spectra, elemental analysis. The IR spectra are taken on a Perki p-E L1eg-325 spectrophotometer in the region of 650–3500 cm-H (NNVD, NUOL). The PMR spectra were recorded on an R-12A (60 MHz) Pivbl Perki n-Elmer spectrophotometer. P D and measure 1. Hydrochloride Sr-pyrolidonmetyl-cysteine (II). To a mixture of 15.7 g (0.1 mol) of cysteine hydrochloride, 12.7 g (0.11 mol of N-hydroxymethyl pyrrolidone and 40 ml of ohlahscene water to 0 ° C) is added with stirring 15 ml of concentrated hydrochloric acid and stirred at at room temperature for 30 minutes, the resulting solution was poured into a mixture of 450 ml of isopropyl alcohol and ether (1: 2), stirred at the onset of crystallization, and the reaction mixture was left in the refrigerator for 4 hours. The product was filtered, washed on the filter with ether and Isotrated from a mixture of methanol and ether. The yield of hydrochloride (I) is 22.95 g (90%, counting on cysteine hydrochloride.) Tpl. 174-175 C, B1 + 4.7 (c1, H, 0), 13.4 (c1, MeOH), Found,%: C 37.70; H 5.92 ; N 10, Calculated,%: C 37.72; H 5.94; M 11.00 PMR spectrum (0.0 + CF5COOH), (f; 8.4 (2H MS, NH), 4, 40 (1H, q,), 3.17 (2H, m,), 4.52 (2H, AH system, 2, 5-SC, MH 3.61 (2H, M, N CH 2.43 (2H, m, NCOCCH), 2.11 m, d (2H, m, OCH2.C). IR spectrum, cm: 3460 (), 1730 (VCO in 5-membered cycle), 1645 ((Mhm amide I), 1515 (ONH, II), 1270 (CTNH, amide III) Example 2. Methyl ester hydrochloride, 3-pyrrolidonemethyl-. -cysteine (II)). To 48 ml of methanol cooled until, under vigorous stirring, dropwise ml of chlorine thionyl is added, then 6 g (0.024 mol) of S-pyridon-cysteine hydrochloride (Cl) is added. and the resulting solution was stirred at 45 ° C for 4 hours, after which it was dissolved in vacuum to dryness. The residue is recrystallized from a mixture of methane ether and dried in vacuo over potassium hydroxide to give 6.27 g (Ml) (99.6% calculated as I). M.p. 70-72 s, oL -5.2 (c1, MeOH). Found,%: C 40.17, H 6.25, N 10, HCl Calculated,%: C 40.22, H 6.38, N 10.42., “DEM spectrum (CRjCOOH) rf: 8.4 ( 2H, 4M.CN-H) -, 4.6 (IH, m,), 4.86 (1H, d, -СНЧ1), 4.45 (2H, K. SOOCH),:. 3.82 (2H, t, NCH2), 3.40 (2H, m,), 2.88 (2H, m, COCH,), 2.35 (2H, m, C-CHi-C), 1 , 40 ppm / S.ZN, t, sn). IR Spectrum, 3420 (); 1730 (-VCO in 5 membered cycle); - 1640 (dNH I); 1520 (cHyN amide I) 1270 (ONH amide IM), 1745. (l) CO el. ether). Example 3.N-tert-butoxycaronyl-5-pyrrolidonemethyl-cysteine (IV). 10.2 g (0.04 mol of S-pyrrolidonemegyl-cysteine (II) hydrochloride) are dissolved by stirring 80 ml of 1N sodium hydroxide solution, and a solution of 12 g (0.055 mol) of di-tert-butyl pyrocarbonate is poured into 40 ml tert. -butyl alcohol. The solution is heated at 45 for 5 minutes, and then stirred at room temperature for 2.5 hours. The reaction mixture is then extracted with 50 mA of petroline ether and the aqueous layer is cooled to 0 ° C and acidified to pH 3- 4 citric acid. The acidified reaction mixture is saturated with sodium chloride (20 g) and extracted with ethyl acetate (3X50 ml) .. Ethyl the acetate extracts are washed with a saturated solution of sodium chloride and then dried over anhydrous sodium sulfate. The ethyl acetate solution was evaporated in vacuo and hexane was added to the residue. The product is recrystallized from a mixture of ethyl acetate and hexane. Output 12.5 g (98%, counting on I), T. pl. 113-114 °, 3 ° С (c 0.5, HjO). Found,%: C 49.10; H 6.97 / ft, 72.. Calculated,%: C 49.04 / H 6.97 / N 8.80. PMR spectrum (DMCQ), ss: .6.90 (1H, d, N11); 4.34 (2H, M, CH2.N); 4.01 (1H, .m,); 3.37 (2H, t,); 2.75 (2H, m, NCOCH-i); 2.00 (2H, m,), 1.43 ppm (p. C (SI)). , IR spectrum, cm: 3330 (l) HH--); 1725 (A) CD in 5 membered cycle); 1695 (DCO urethane); 1640 ((fNH amide I); 1530 (cH NH amide II); 1265 (dNH amide III). Example 4. Methyl ester M-tert-butoxycarbonyl-5-pyrrole and -methyl-cysteinyl-S-pypropyldimethyl-cysteine (V To cooled to -15 - 20 With rast: a thief 1.27 g (4 mmol) of N-tret-botoxicarbonyl-S-pyrrolidonmethyl-L-cysteine in 5 ml of dimethylformamide is added dropwise with stirring 0.44 ml (4 mmol) 14- methyl morpholine and then 0.52 ml (4 mmol) of chlorobolic acid isobutyl ester. After 10 minutes, a cooled mixture of 1.075 g (4 mmol) of 5-pyrrolidonmethyl-1-cysteine methyl ester hydrochloride and 0.44 ml (4 mmol) of N- methylmorpholine B 5 MP di methylformat | 1ida and nepehie are sprinkled for another 3.5 hours at -15 -, after the reaction is completed, the mixture is diluted with water and extracted with ethyl acetate. The organic layer is taken in succession with 0.25 sodium bicarbonate solution, 0.05 N sulfuric acid, and The solvent was evaporated in vacuo and hexane was added to the residue. The resulting product was recrystallized from a mixture of ethyl ester of acetic acid and hexane. The yield of 1.92 g (90%), so pl. .78-80 s (. 47, (c 0.1, MeOH). Found,% t С 49.54; H 6.94, N10.4 С2аНз М4075. Calculated,%: С 49.61; H 6 , 81, N 10.52. Spectrum PMR „(CO C1z): 5.4 (1H, d, NH); 5.26 (1H, D, NH); 4.52 (1H, m); 4 , 34 (4n, m, u) 3.70 (IH, m, Cf-H); 3.58 (3N, s, -, СООСНа); 3.02 (4H, t, NCH o and NCH), 2 , 90 (4H, m, CPHG and CPHg), 2.4 (4H, m, CH-L-CO and CH1CO); 2.0 (4H, M, C-CHi-C and C-CIU-C); 1.42 ppm (9H, s, C (CH, 4 IR spectrum, cm: 3420 (1 N H), 173 (l) CO in the 5-membered cycle), 1745 (VCO el. Ether); 1695 (Deo urethane); 1640 amide I), 1530 (rfNH amide II), 12-70 cGm H amide II). Example 5. Cyclic disulfide of methyl ester of N-tert-butoxycarbonyl-cysteinyl-cysteine (VI). To a solution of 1.27 g (5 mmol) of iodine in 50 ml of methanol, a solution of 0.67 g (1.25 mmol of methyl ester of Boc-S-pyrrolyronmethyl-cysteinyl-5-pyrrolidone-methyl-Cysteine (V) in 30 ml of methanol, after which the mixture is stirred for an additional 40 minutes After cooling, 1N sodium thiosulfuric acid solution is added until the yellow color is obtained, the mixture is evaporated with dl iu MP, washed with 0.5N sodium sulfate and sodium sulfate and water, dried over anhydrous sodium sulfate. Dissolve; ts1 is evaporated, the residue is crystallized out when adding petroleum Lane ether. The yield after recrystallization from Hlyuroforma mixture of petroleum ether and 0.39 g, measured as the IV). M.p. 186-187s, 67.8 (c 1, MeOH). Total yield 73% (per cysteine hydrochloride). Found,%: C 42.97; H 6.04, N3.30 Ql HjoNj Og-S / i. Calculated,%: C, 42.89 / H, 5.99; N 8.33. Literary data: So pl. 185187, 64, (with 1.87, MeOH). Found: C, 43.13; H 6.21; N 8.38. 180-183 2. Formula of the Invention Derivatives of S-pyrrolidone-methyl-ristein with the general formula X-NH-H-CO-Y. SI, S-CH2-NCr3 where X is (CH.) ZSOCO (Woe) or NSP Y is OH or OCH3, .., -pyrrolidonmethyl- (Ruth), O. as the intermediate products of the synthesis of cystine-containing peptides. Sources of information taken into account in the examination 1. US Patent 3560521, cl. 260-326.3, pub. 1971. 2. The patent of Germany 2060969, class. 129 a - § / OlJ - Publ. 1971. Arsh G.Sh., Papekhevich OS, Mixta S.Ya. 5-benzamide solvents and derivatives of L-cysteine and-mercapto propionic acid, JO, 1975, 45, 6, p. 1384-1388. 4. Chemistry of polypeptides. Ed. f. Kahso nisa. Per. from English, M., world, 1977.