RU2043337C1 - Method of synthesis of 5-arginylaminonaphthalene-1-sulfamides - Google Patents

Abstract

FIELD: organic chemistry. SUBSTANCE: process is carried out by interaction of N_q-nitro-N_α-tert. -butylhydroxycarbonylarginine with 5-amino-naphthalene-1-sulfamide in the form of base in pyridine and dioxane medium in the presence of di-tert.-butylpyrocarbonate, and compound

is obtained where values R¹, R² are given above. Protective groups were removed with hydrogen in hydrogen chloride solution in methanol in the presence of Pd/C. Method provides yield increase from 35-40% to 49-62.7% EFFECT: increased yield of product. 7 tbl

Description

где R¹ H, R² CH₃; C₂H₅; н-, i= C₃H₇; н-, i-, t-C₄H₉; C₅H₁₁; C₈H₁₇;

; -CH

или R¹ R² CH₃;
C₂H₅; C₃H₇; н-, i-C₄H₉; или
NR¹R²=

O;

;

которые применяют в биотехнологии в качестве хромо- и флюорогенных субстратов при определении ферментативной активности протеаз и анализа сложных их смесей, а также могут быть использованы для синтеза других производных на их основе, применяемых для таких же целей.The invention relates to bioorganic chemistry, specifically to an improved method for producing arginyl derivatives of aminonaphthalene sulfamides of the general formula
Argnh

where R ¹ H, R ² CH ₃ ; C ₂ H ₅ ; n-, i = C ₃ H ₇ ; n-, i-, tC ₄ H ₉ ; C ₅ H ₁₁ ; C ₈ H ₁₇ ;

; -Ch

or R ¹ R ² CH ₃ ;
C ₂ H ₅ ; C ₃ H ₇ ; n-, iC ₄ H ₉ ; or
NR ¹ R ² =

O;

;

which are used in biotechnology as chromo- and fluorogenic substrates in determining the enzymatic activity of proteases and analysis of their complex mixtures, and can also be used for the synthesis of other derivatives based on them, used for the same purposes.

The closest is a method for producing 5-arginylaminonaphthalene-1-pentamethylene sulfamide, in which N ^α- carbobenzoxy arginine hydrochloride is mixed with N-hydroxybenzotriazole (OBT) in dimethylformamide (DMF) and condensed with 5-aminonaphthalene-1-pentamethylenediamine dimethyl dimethyl dimethyl dimethyl dimethyl dimethyl dimethyl dimethyl amide the resulting HBr salt of N ^α -protected arginyl-ANSA decompose and remove the protective carbobenzoxy with hydrogen bromide in glacial acetic acid, HBr salt isolated arginyl-ANSA, it is decomposed and highlight t product as a free base in 40% yield disadvantage of the method-prototype is the low yield of the desired product (≈40% based on the starting) and multistage. In addition, the method uses a strong allergen, dicyclohexylcarbodiimide, not produced in the CCCP industry, hydrogen bromide in glacial acetic acid, and a product (by-product) of benzyl bromide, which is a difficult to utilize lacrimator, is formed. The possible isolation of the target products in the form of bromohydrates is impractical, since they are not stable during storage and are not convenient for further use due to the reactivity of HBr (it can restore the sulfamide group, and the other labile groups formed in this process oxidize Br ₂ ).

 The aim of the invention is to increase the yield of target products.

The goal is achieved by a method including obtaining N-protected 5-arginylaminonaphthalene-1-sulfamide, removing the protective group and isolating the target product, in which 5-aminonaphthalene-1-sulfamide is used as the starting compound in the form of a free base, it is acylated with N ^q -nitro -N ^α -tert. Butyloxycarbonyl-arginine in pyridine-dioxane medium in the presence of di-tert-butylpyrocarbonate to obtain 5-N ^q -nitro-N ^α -tert. Butyloxycarbonyl) arginylaminonaphthalene-1-sulfamide, and the deprotection is carried out hydrogenation in astvore methanolic hydrogen chloride in the presence of a palladium-carbon catalyst to give the dihydrochloride 5-arginilaminonaftalin-1-sulphonamide.

Distinctive features of the proposed method are, firstly, the use of a specific feedstock, as well as the change of other reagents and actions inherent in the previously known method, with a positive effect. In the proposed set of operations, cleaner intermediates are formed, stages are reduced, the yields of the target products are increased (49.0-62.7%) based on the reaction N ^q -nitro-N ^α -tert.butyloxycarbonyl-arginine (see table 7). In addition, the obtained intermediate products in the form of 2 НСI-salts of arginine-ANSA are convenient for long-term storage due to their stability, and, if necessary, can be decomposed to free bases in high yields.

 PRI me R 1 (synthesis of the target product according to the prototype). 5-arginylaminonaphthalene-1-pentamethylene sulfamide.

A solution of 3.5 g (10 mmol) of N ^α -karbobenzoksiarginina in 20 ml of dimethylformamide was surged under stirring at -4 ^{° C} to 1.33 g of 1-hydroxybenzotriazole in 10 ml of dimethylformamide was added 2.1 g of dicyclohexylcarbodiimide and stirred for 40 min at 0 ^C. to this solution was added 3.7 g (10 mmol) of hydrobromide of 5-aminonaphthalene-1-pentametilensulfamida and 2.8 ml of triethylamine. The mixture was stirred for 48 hours, filtered, 60 ml of ethyl acetate and 120 ml of hexane were added to the filtrate, decanted, the precipitate was washed with ethyl acetate-hexane, dissolved in butanol-water, the upper layer was separated, the aqueous was extracted with butanol, the combined butanol extracts were washed with 5% aqueous NaNO ₃ and aqueous KHSO ₄ , water and evaporated to dryness. The resulting preparation (R _f = 0.75, butanol-acetic acid water 4: 1: 2) was introduced into the next synthesis step without crystallization.

4 g of the N ^α -protected derivative was stirred for 2.5 hours at ²⁰ ° C with 40 ml of 2N. HBr in CH ₃ COOH, poured into 600 ml of ether with stirring, the precipitate was filtered off, dissolved in 200 ml of water, extracted with ethyl acetate, the aqueous layer was evaporated to dryness, the residue was dissolved in butanol-CH ₃ COOH-water (4: 1: 2) and chromatographed on a column (5 x 15 cm) with silica gel in the same system. Area of blue fluorescence were collected, evaporated to dryness, dissolved in 200 ml of water was stirred ^at 50 ° C with 0.5 g of charcoal, filtered, to the filtrate was added 1 g of NaBr, was evaporated to 20 ml, pH adjusted to 7.8 with Na ₂ CO ₃ and left at 4 ^about C. The residue was separated, dried in vacuum. Yield 2.1 g (40%). R _f = 0.28 (in the same system, on silofol plates). It is manifested by ninhydrin on TLC and has blue (λ = 440 nm) fluorescence upon UV irradiation.

PRI me R 2. 5- (N ^q -nitro-N ^α -tert. Butyloxycarbonyl) arginylaminonaphthalene-1-pentamethylene sulphamide.

3.2 g of (N ^q -nitro-N ^α -tret.butiloksikarbonil) arginine, 0.8 ml of dry pyridine and 2.4 ml of di-tret.butilpirokarbonata stirred in 10 ml of dry dioxane at ²⁰ ° C for 1.5 am. 2.9 g of 5-aminonaphthalene-1-pentamethylene sulfamide are added in small portions over the course of 50 minutes to the resulting solution. The reaction mixture was stirred for 4 hours, another 1.6 ml of di-tert-butyl pyrocarbonate was added and stirred for 17 hours. Then the dioxane was evaporated under reduced pressure, the oily residue was dissolved in acetone and the solution was passed through an alumina chromatography column, eluted with ethyl acetate-methanol ( 1: 1). The fraction containing the target product is boiled for 0.5 hours with activated carbon, filtered off, and the filtrate is evaporated. The oily residue was triturated with dry diethyl ether, the resulting residue was filtered off and dried in air. 3.73 g (63%) of chromatographic and analytically pure product are obtained, mp. 117-121 ^{° C,} R _f = 0,70 on Silufol plates (ethyl acetate-methanol 19: 1), α | _D ²⁰ - 5,8 (CI; acetone).

 Found C, 52.77; H 6.51; N, 16.71; S 4.83.

C ₂₆ H ₃₇ N ₇ SO ₇ .

 Calculated, C 52.78; H6.30; N, 16.58; S 5.42.

 Similarly, other N-protected 5-arginylamino-naphthalene-1-sulfamides were obtained. Their yields are given in table 1, physico-chemical properties in table 2.

 PRI me R 3. 5-Arginylaminonaphthalene-1-pentamethylene sulfamide dihydrochloride.

5.92 g of 5- (N ^q -nitro-N ^α -tert-butyloxycarbonyl) arginylaminonaphthalene-1-pentamethylene sulphate was dissolved in 150 ml of dry methanol, 10 ml of 4.4 N was added. HCl in methanol and 2 g of catalyst (10% Pd / C). Was stirred in a hydrogen stream of 25 hours at 20 ° ^C, add an additional 2.5 g of catalyst and 4.5 ml of HCl in methanol, stirred for another 23 hours in a stream of hydrogen. After this, the reaction mixture was filtered, the filtrate was evaporated, the oily residue was triturated with dry diethyl ether, filtered and washed with ether. Obtain 4.73 g (91%) of the target product, so pl. 141-145 ^о С, R _f = 0.40 (on the silofol plates n-butanol-acetic acid-water 4: 1: 2), α | _D ²⁰ + 6.3 (CI; CH ₃ OH). 1 H-NMR (DMSO): 1.38 (CH ₂ ), 3.06 (CH ₂ ).

 Found C, 48.76; H 6.45; N, 15.94; S 5.46; CI 13.24.

C ₂₁ H ₃₂ N ₆ SO ₃ Cl ₂ .

 Calculated, C 48.55r; H 6.21; N, 16.18; S 6.17; Cl 13.65.

 Similarly, the removal of the protective group of other N-protected arginylaminononaphthalene sulfamides is carried out. The yield of the corresponding dihydrochlorides obtained is given in table. 3, their physicochemical properties in table 4.

 PRI me R 4. 5-Arginylaminonaphthalene-1-pentamethylene sulfamide.

5.2 g (0.01 mol) of 5-arginylaminonaphthalene-1-pentamethylene sulfamide dihydrochloride are dissolved in 50 ml of water, 200 ml of n-butanol are added to the solution in a separatory funnel and (in 20 ml portions, shaking after each portion) 5 % aqueous NaHCO ₃ to pH> 7 (aqueous layer). The organic layer is washed with water twice in 20 ml and evaporated to 10-15 ml. The residue was triturated with 150 ml of dry ether, filtered off, washed with ether, and dried. 4.15 g (93%) of the expected product are obtained. Mp 138-143 ^{° C,} R _f 0,40 (same sistema.α | _D ²⁰ + 6,9 (Cl; CH ₃ OH).

 Found, C 56.24; H 6.77; N, 18.85; S 6.54.

C ₂₁ H ₃₀ N ₆ SO ₃ .

 Calculated, C 56.48; H 6.77; N, 18.82; S 7.18.

 Analogously to example 4, other 5-arginylaminonaphthalene-1-sulfamides were obtained. Their yields are given in table 5, physico-chemical properties in table 6.

The yield of final products in terms of the amount of the starting compound (N ^q -nitro-N ^α -tert. Butyloxycarbonyl) -arginine (3.2 g) taken in the synthesis is shown in Table 7.

As can be seen from the above examples and table. 7, the proposed method allows to increase the yield of the final product from 40% to 49.0-62.7%. An additional technological advantage of the proposed method is the ability to isolate stable intermediate products of arginylaminonaphthalene sulfamide dihydrochlorides, which, unlike (N ^α -carbobenzoxy) arginylaminonaphthalene sulfamides, arginylaminonaphthalene sulfamides and their bromohydrates can be stored for a long time and, if necessary, directly used in peptide synthesis without undesirable side reactions.

 Using the invention allows to obtain arginyl-ANSA on an industrial scale from available raw materials and with high yields.

Claims (1)

METHOD FOR PRODUCING 5-ARGINYLAMINONAFTHALIN-1-SULFAMIDES of the general formula

where R ₁ H; R ₂ CH ₃ , C ₂ H ₅ ; n-; iC ₃ H ₇ ; n-, i-, tC ₄ H ₉ ,

C ₅ H _{1 1} ; C ₈ H _{1 7} ;
or
R ₁ = R ₂ CH ₃ , C ₂ H ₅ , C ₃ H ₇ , n-, iC ₄ H ₉ ,
or

the interaction of N protected arginine with the corresponding aminonaphthalene sulfamide in an organic solvent in the presence of a condensing agent to obtain N-protected 5-arginylaminonaphthalene -1-sulfamide by its isolation, deprotection and isolation of the target product, characterized in that, in order to increase the yield of the product, As N protected arginine, N ^q nitro N ^α tert-butyloxycarbonyl arginine is used, which is reacted with 5-aminonaphthalene-1-sulfamide in the form of a base, where as a solution Pyridine and dioxane are used, and di-tert-butyl pyrocarbonate is used as the condensing agent to obtain N-protected 5-arginylaminonaphthalene-1-sulfamide of the general formula

where R ₁ and R ₂ have the indicated meanings, and by deprotection with hydrogen in a solution of hydrogen chloride in methanol in the presence of palladium on carbon as a catalyst.

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