RU2054001C1 - Method of synthesis of 5-arginylaminonaphthalene-1-sulfamides - Google Patents

Abstract

FIELD: organic chemistry. SUBSTANCE: product: 5-arginylaminonaphthalene-1-sulfamides of the formula

where R₁=H; R₂=CH₃, C₂H₅, C₃H₇, iso-C₃H₇, C₄H₉, iso-C₄H₉, C₅H₁₁, C₆H₁₁; R₁=R₂=CH₃, C₂H₅, NR₁R₂ together

; empirical formulas are БФ C₂₁H₃₀N₆SO₃, C₁₇H₂₄N₆SO₃, C₁₈H₂₆N₆SO₃. Reagent 1: N^α-tert.-butylhydroxycarbonylarginyl hydrochloride (HCl). Reagent 2: 5-aminonaphthalene-1-sulfamide. Reaction conditions: in the medium of dimethylformamide (DMFA) and pyridine, in the presence of di-tert.-butylpyrocarbonate followed by protecting group removal. EFFECT: improved method of synthesis. 3 cl, 3 tbl

Description

где при R₁=Н; R₂=CH₃, C₂H₅, C₃H₇, i=С₃Н₇, С₄Н₉, i-С₄Н₉, С₅Н₁₁,

или при R₁= R₂=СН₃, С₂Н₅, или NR₁R₂= -N

O -N

,-N

, которые применяются 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 [1]
Наиболее близким является способ получения 5-аргиниламинонафталин-1-пентаметиленсульфамида, исходя из N^α-карбобензоксиаргинина гидрохлорида, который перемешивают с N-гидроксибензотриазолом (ОВГ) в диметилформамиде (ДМF) и конденсируют с 5-аминонафталин-1-пентаметиленсульфамида бромгидратом в присутствии дициклогексилкарбодиимида и триэтиламина, образовавшуюся НВr соль N^α-защищенного аргинил-АНСА разлагают и защитную карбобензоксигруппу снимают бромистым водородом в ледяной уксусной кислоте, образовавшуюся при этом НВr соль аргинил-АНСА очищают, разлагают и выделяют продукт в виде свободного основания с выходом 40% [2]
Недостатком способа является низкий выход целевого продукта, а также многостадийность, использование сильного аллергена дициклогексилкарбодиимида и непроизводимого в нашей промышленности бромистого водорода в ледяной уксусной кислоте, образование побочного продукта бромистого бензила, который является трудноутилизируемым лакриматором (НВr/СН₃СООН сильная кислота, разрушает аминонафталинсульфамиды). Кроме того, возможное выделение целевых продуктов в виде бромгидратов нецелесообразно, так как они неудобны в дальнейшем использовании вследствие реакционноспособности НВr (он может восстановить сульфамидную группу, а образующийся при этом Br₂ окислить другие лабильные группы). Свободные основания неудобны в тех случаях, когда необходимо их длительное хранение (из-за их нестабильности). Перечисленные факторы в совокупности являются значительным препятствием для получения аргиниламинонафталинсульфамидов и субстратов на их основе в промышленных масштабах.The invention relates to bioorganic chemistry, specifically to an improved method for producing arginyl derivatives of aminonaphthalene sulfamides of the general formula

where at R ₁ = N; R ₂ = CH ₃ , C ₂ H ₅ , C ₃ H ₇ , i = C ₃ H ₇ , C ₄ H ₉ , i-C ₄ H ₉ , C ₅ H ₁₁ ,

or when R ₁ = R ₂ = CH ₃ , C ₂ H ₅ , or NR ₁ R ₂ = -N

O-n

, -N

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 [1]
The closest is a method for producing 5-arginylaminonaphthalene-1-pentamethylene sulfamide, starting from N ^α- carbobenzoxy arginine hydrochloride, which is mixed with N-hydroxybenzotriazole (OGH) in dimethylformamide (DMF) and condensed with 5-aminonaphthalene-1-pentamethylene sulfamide dimethyl dimethyl sulfamide bimide triethylamine, the formed HBr salt of the N ^α -protected arginine ANSA is decomposed and the protective carbobenzoxy group is removed with hydrogen bromide in glacial acetic acid, the resulting HBr salt of arginine ANSA purify, decompose and isolate the product in the form of a free base with a yield of 40% [2]
The disadvantage of this method is the low yield of the target product, as well as multistage, the use of the strong allergen dicyclohexylcarbodiimide and hydrogen bromide that is not produced in our industry in glacial acetic acid, the formation of benzyl bromide by-product, which is a difficult to utilize lacrimator (HBr / CH ₃ COOH strong acid, destroys aminone sulfide) . In addition, the possible isolation of the desired products in the form of bromohydrates is not practical, since they are inconvenient in future 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 ). Free bases are inconvenient in those cases when they need long-term storage (due to their instability). These factors together are a significant obstacle to the production of arginylaminonaphthalene sulfamides and substrates based on them on an industrial scale.

 The aim of the invention is to increase the yield, as well as simplifying the process by reducing stages.

The goal is achieved by using the acylation reaction using N ^α -protected arginine hydrochloride, deprotecting, neutralizing and isolating the product, the free base of 5-aminonaphthalene-1-sulfamide being acylated, N ^α -t-butyloxycarbonylarginine is used as protected arginine, the process is carried out in mixtures of dimethylformamide-toluene in the presence of an excess of di-tert-butyl pyrocarbonate and pyridine and without isolating the resulting N ^α -t-butylosarbonyl-arginyl-ANSA remove the protective group by treatment with a reaction mixtures of trifluoroacetic acid in methylene chloride.

 The method differs in conditions ensuring the process with the formation of the target product, namely the use of certain starting materials and reagents, therefore, according to paragraph 9.02. Instructions EZ-2-74 is an object of the invention. In the proposed set of technological operations, cleaner intermediate products are formed and there is no need for their isolation, as well as labor-intensive product purification, including chromatography, staging is reduced and the yield of the target products is increased (they reach 62-84%). In addition, it is possible to directly obtain the target products in the form of HCl salts. The latter (if necessary) decompose to a free base with a high yield.

PRI me R 1. 5-Arginylaminonaphthalene-1-pentamethylene sulfamide. 3.3 g of N ^α -tert-butiloksikarbonilarginina (monohydrate) dissolved in 10 ml of dimethylformamide is poured 30 ml of dry toluene. The solution was evaporated under reduced pressure to 10 ml, cooled ^to 10 ° C, is added 0.8 ml of dry pyridine and 2.7 ml of di-tert butylpyrocarbonate. It is stirred for 1 hour, 3.2 g of 5-aminonaphthalene-1-pentamethylene sulfamide are added portionwise over 40 minutes. Stirred for 3 hours at ²⁰ ° C after which was added 1.3 ml of di-tert-butylpyrocarbonate and stirred for a further 16 hours. The reaction mixture was poured into 100 ml of dry ether, stirred for 10 minutes, the ether was decanted, the oily residue was poured 50 ml of trifluoroacetic acid methylene chloride (1: 1). The reaction mixture was stirred for 1 hour at ²⁰ ° C, concentrated to 10 ml and poured 100 ml of dry ether. The resulting product is filtered off, dried in a vacuum desiccator, dissolved in 20 ml of water, and unreacted aminonaphthalene sulfamide is filtered off. The aqueous solution was extracted with 80 ml of n-butanol, the organic layer was separated, washed with 5% aqueous NaHCO ₃ to pH 7-8 (aqueous layer), then with water (twice in 10 ml). The solvent was evaporated under reduced pressure; the oily residue is triturated with ether, the resulting crystalline precipitate is filtered off and dried. 2.80 g (62%) of chromatographic and analytically pure product are obtained. Mp 138-143 ^about With, R _f = 0,41, (butanol CH ₃ COOH water 4: 1: 2), / α / $\genfrac{}{}{0ex}{}{\text{20}}{\text{D}}$ +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.

 Other products of formula 1 are prepared in a similar manner; the conditions are given in table. 1, the data of elemental analysis of the target products in table. 2.

PRI me R 2. Dihydrochloride 5-arginylaminonaphthalene-1-pentamethylene sulfide. The treatment of the reaction mixture to the stage of neutralization inclusively is carried out in the same manner as in example 1; after neutralization with NaHCO ₃ and washing, the mixture was concentrated to 15 ml and saturated with dry HCl. Further, the selection of the product is continued as in example 1. Yield 2.96 g (57%). Mp 141-145 ^о С, R _f = 0.40 (the same system), / α / $\genfrac{}{}{0ex}{}{\text{20}}{\text{D}}$ + 6.3 (Cl; CH ₃ OH). 1 H-NMR (DMSO): 1.38 (CH ₂ ), 3.06 (CH ₂ ).

Similarly to examples 1 and 2 receive other dihydrochlorides. For example, 5-arginylaminononaphthalene-1-cyclohexyl sulfamide dihydrochloride is obtained from 3.3 g of 5-aminonaphthalene-1-cyclohexyl sulfamide 4.4 g (82%). R _f = 0.42 (same system) / α / $\genfrac{}{}{0ex}{}{\text{20}}{\text{D}}$ +17.7 (Cl; CH ₃ OH). PMR (DMSO): 1.23 (CH ₂ ); 3.20 (CH).

 The characteristics of the remaining dihydrochlorides Arg-ANSA are given in table. 3.

PRI me R 3. 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 serving) 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.

 Thus, the method allows to increase the yield of target products in comparison with the known from 40 to (61-84)% and reduce the staging (from 4 stages to 2), eliminate toxic reagents (dicyclohexylcarbodiimide, hydrogen bromide in glacial acetic acid) and side product (benzyl bromide).

Claims (2)

1. METHOD FOR PRODUCING 5-ARGINYLAMINONAFTALIN-1-SULFAMIDES of the general formula

where at R ₁ -H, R ₂ -CH ₃ , C ₂ H ₅ , C ₃ H ₇ , iso-C ₃ H ₇ , C ₄ H ₉ ; iso-C ₄ H ₉ ;

or when R ₁ = R ₂ - CH ₃ , C ₂ H ₅ ,
or N, R ₁ R ₂ together -

by the interaction of N-substituted arginine hydrochloride with the corresponding 5-aminonaphthalene-1-sulfamide in an organic solvent in the presence of a condensing agent, followed by removal of the protective group and isolation of the target product, characterized in that N ^α -tetra hydrochloride is used β-butyloxycarbonyl arginine, 5-aminonaphthalene-1-sulfamide are used in the form of a base, dimethylformamide and pyridine are used as a solvent, and diter is used as a condensing agent. butyl pyrocarbonate, deprotection is carried out with a solution of trifluoroacetic acid in methylene chloride. 2. The method according to claim 1, characterized in that the hydrochloride N ^α -tert-butyloxycarbonyl arginine and di-tert-butyl pyrocarbonate are taken in a molar ratio of at least 1: 1.8, respectively.

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