SU791744A1 - Dimethylester of 4-phthalimido-n-phthalyl-dl-glutamic acid as intermediate product for synthesis of erythro-4-amino-dl-glutamic acid - Google Patents

Description

(Vl). The total yield of the substance, g, calculated on the starting diazo acetic ester (or methyl acrylate), is 32%.

The disadvantage of this method is the complexity and complexity of the process carried out at high pressure (246 atm), explosive (hydrogen, diazo acetic ester and highly toxic substances such as diazomethane, benzoyl chloride) are used.

The aim of the invention is the synthesis of 4-phthalimido-M-phthalyl-01-glutamic acid dimethyl ester, which is an intermediate product for the synthesis of 4-amino-01-glutamic acid spectrometry.

This is achieved thanks to the new chemical structure of dimethyl ester 4-phthalmicide-M-phthalyl-01-glutamine CH-COOCH,

1-5

Bg

about

(these conditions using 4-bromo-N-phthalyl-1.-glutamic acid dimethyl ester occur almost completely racemizing the product. Target product (1) is isolated from the reaction mixture as a mixture of diastereoisomers by conventional methods, for example by column chromatography on silica gel. 4-phthalimido-M-phthalyl-01-glutamic acid dimethyl ester is a colorless, crystalline, optically inactive substance, mp 150-170 ° C. The wide melting range is due to the fact that substance is a mixture of two diastereisomers. Chromatographically it appears as two closely spaced spots having R -0.48 and 0.55 in the system chloroform-benzene-methanol (12: 10: 1) on platinum Silufoluv -25 type. The yield of 4-phthalimido-N-phthalyl-I Ju-glutamic acid dimethyl ester by the proposed method is 59%, based on 4-bromo-H-phthalyl-01- (or 1) -glutamic acid dimethyl ester The structure of 4-phthalimido-N-phthalyl-01-glutamic acid dimethyl ester was confirmed by elemental analysis, UV and IR spectroscopy. In the UV spectrum there are two absorption maxima: at 220 and 298 nm, confirming the presence of phthalic groups, In the IR spectrum there are the following absorption bands, indicating the presence in the compound (1) of a functional acid, which is expressed by the formula

Hjf OOC-CH-CH, -CH-COOCH, II

Ivl n

0 Vo 0

The method of producing dimethyl ester is based on the known reaction of replacing a halogen atom with a phthalimido group and consists in the fact that 4-bromo-M-phthalyl-01 (or L) -glutamic acid dimethyl ester is reacted with potassium phthalimide in an organic solvent medium, mainly of methyl formamide, at a temperature of 80-100 C for 4-5 h according to the scheme:

HjCOOd-CH-CH - CH.-COOCHj

.w ij

/five

In groups: 1780 and 1720 cm phthalimide groups, 1740 sms 0 ester groups. Example. 3.84 g of dimethyl 4-bromo-N-phthalyl-DL- (or L) -glutaminebvoic acid and 3.7 g of phthalimide porridge are poured into 35 ml of dimethylformamide (DMF) and the mixture is heated at 80 ° C for 4 hours. The precipitate is filtered off and the DMF is evaporated under reduced pressure. To the residue was added 60 mp of chloroform, filtered, the filtrate was washed with water, 10% NaOH solution and again twice with water. The chloroform solution is dried with sodium sulfate, evaporated, and dimethyl 4-phthalimido-N-phthalyl-DL-glutaminic acid dimethyl ester is isolated by column chromatography. A 150 x 2 column (cm) filled with silica gel L 40/100 is used. Elute with chloroform-benzene-methanol (12: 10: 1). Fractions i are collected, having 55 and 0.48 in this system on Silufol UV-254 plates (appearance in UV light), combined and evaporated in vacuo. 4-phthalimido-M-phthalyl-01-glutamic acid dimethyl ester is obtained, which is a colorless crystalline product, mp 150-170 ° C. Yield 2.67 g, which corresponds to 59% of the theoretically calculated. Found,%: C 61.44, H 3.98, N 6.30 ClbHia OeNi Calculated: C 61.30, H 4.00, N 6.23.

UFS1nm) 220, 298-M-phthalic corpse

IR (cm) 1740-CO-esters of 1780,1720-CO-phthalimide groups.

The proposed method for the preparation of 4-phthalimide-H-phthalyl-oi-glutamic acid dimethyl ester provides a number of advantages compared to the method for producing a compound, the multistage is eliminated, high pressure is not used, and therefore the equipment design is simplified, and there is no need to use explosive ones. and highly toxic substances such as hydrogen, diazomethane, diazo acetic ester, etc.).

4-phthalimido-M-phthalyl-01-glutamic acid dimethyl ester is used to prepare erythro-4-amine-DL-glutamic acid, which has the ability to inhibit the glutamine synthetase enzyme.

For this purpose, dimethyl erythro-4-phthalimido-M-phthalyl-OL-glutamic acid dimethyl ester is isolated from dimethyl 4-phTcLPi-Imido-N-phthalol-D-L-glutamic acid (1), a mixture of diastereisomers, by adsorption chromatography mainly on silica gel.

Erythro-4-phthalimido-M-phthalyl-OL-glutamic acid dimethyl ester is further subjected to hydrolysis in the presence of mi: neutral acid, predominantly saline by boiling the reaction mass for 5-6 h, followed by treatment of the resulting erythro-4 dihydrochloride α-amino-oi-glutamic acid in the usual manner, for example, pyridine in an alcohol-aqueous medium. An erythro-4-amino-01-glutamic acid, which is a colorless crystalline substance, is obtained, mp. 270C causes a positive reaction with ninhydrin. The data of elemental analysis and IR spectroscopy confirm the composition and structure of such a compound.

Erythro-4-amino-1) b-glutamic acid is not described in the literature.

Acid has an inhibitory effect on the enzyme glugamin synthetase, the key enzyme for nitrogen metabolism in plants and animals. One of the most important reactions catalyzed by this enzyme is the biosynthesis of gummin, which plays an indispensable role in the construction of bases of nucleic acids — any cell is fundamentally based. As a result, glutamine synthetase enzyme inhibitors can exhibit a variety of biological activities — antitumor, antiviral, etc.

A specific feature of erythro-4-amino-0L-glutamic acid is the ability to inhibit the glutamine synthetase enzyme in the presence of magnesium and manganese, the percentage of inhibition is about 30% in either case. In addition, it exhibits a low general toxicity when administered intraperitoneally to animal mice), while

0 as the most potent inhibitor of glutagne synthetase, methionine-Bb-sulfoximine, is an order of magnitude more toxic (mg / kg) - its toxic effect is manifested in damage to the central nervous system, and therefore is not used in medical practice.

Claims (1)

Invention Formula 4-phthalimido-N -phthalyl-1) -L-glutamic acid dimethyl ester of the formula HjCOOC - CH-SK-CH coaxially -№-N ° OGG O yo as an intermediate for the synthesis of erythro-4-amino-11b-glutamic acid. Sources of information taken into account during the examination 1. E. Carter, F.R. Van Abeele, W. Rothrock, J. Bio). Chim, 178, c ..325334, 1949.