RU2038095C1 - Method for producing glucosamine hydrochloride possessing antiarthrosis activity - Google Patents

Abstract

FIELD: medicine. SUBSTANCE: remedy is individual substance of D- glucosamine hydrochloride obtainable in chitin hydrolysis with concentrated hydrochloric acid, chitin being utility waste in shrimp and krill processing. Method involves carrying out chitin hydrolysis with concentrated hydrochloric acid (1: 10) during 2 h at water bath temperature of 75 C. Hydrolysate is thinned with water. Hydrolysis is performed together with purification with activated carbon at 60 C. The hydrolysate is boiled down in vacuum with temperature being not higher than 60 C until the residual liquid quantity is 15% of initial hydrochloric acid quantity. The target product is separated by its crystallization from acid water solution in continuous stirring up. EFFECT: enhanced purity; enhanced effectiveness in treating heavy cases of arthroses. 1 tbl

Description

 The invention relates to the pharmaceutical industry and relates to a method for the production of an anti-arthritic drug from chitin obtained from secondary raw materials from processing shrimp, krill and other crustaceans mined by the fishing industry.

 There is a method of producing an anti-arthritic agent D-glucosamine sulfate from the shell of crabs and shrimps by treating the raw material with hydrochloric acid, converting the chloride to the base, treating the latter with sulfuric acid to form glucosamine sulfate (Swiss Patent No. 525861).

 It is known that D-glucosamine sulfate has an anti-arthronic effect, but it is not very stable, hygroscopic, and easily oxidized, especially when wet. Its stabilization is carried out by adding two moles of sodium chloride per mole of substance.

 The disadvantage of this drug is the high content of sodium chloride (about 13%), which leads to an increase in the irritating effect on living tissue with intramuscular injection of the drug itself with a 20% solution.

 Known methods for producing D-glucosamine hydrochloride from the shell of crustaceans with a yield of 50-55% of the original chitin with a purity of 98% also with a yield of 48% of chitin.

A known method of producing a reagent D-glucosamine hydrochloride from crab meal by obtaining and subsequent hydrolysis of chitin with conc. Hydrochloric acid (1: 10) in a boiling water bath (100 ^about C) for 2.5 hours, adding water (1: 100 chitin- water), bleaching with activated carbon (140: 50 chitin-charcoal) at 60 ° ^C for 20 minutes, filtration, evaporation in vacuo at 10-12 mm Hg and a temperature of 60-70 ^{° C,} precipitation with ethyl alcohol (140: 80 chitin-alcohol) and crystallized overnight (13-15 h), washing with ethanol (140: 20 chitin-ethanol), push-ups, and drying in a vacuum desiccator over calcium chloride. Yield 61-64% of chitin. The described method is close to the claimed and selected for the prototype.

 The disadvantages of this method are: low yield (64%) of the target product, hydrolysis at high temperature leads to the destruction of part of the substance, the stage of precipitation with alcohol leads to the formation of a fine crystalline precipitate, and therefore, more heavily contaminated and difficult to filter. A small amount of alcohol taken to wash the precipitate does not allow complete removal of impurities and by-products of hydrolysis, as well as to wash the precipitate from concentrated hydrochloric acid.

 We did not find in the literature information on the anti-arthritic effect of D-glucosamine hydrochloride. However, there is evidence of the incorporation of glucosamine into the macromolecule of nerve endings and the conversion of glucosamine to glucose and aminogluconic acid using animal tissue.

 The aim of the invention is to increase the yield of the target product, the exclusion of alcohol at the stage of crystallization of the product, the expansion of the arsenal of medical devices for the treatment of arthrosis.

This goal is achieved in that the method of producing D-glucosamine hydrochloride for medical purposes, with a purity of 99%, is carried out by the stepwise hydrolysis of chitin at a low temperature. Initially, the hydrolysis carried out with concentrated hydrochloric acid (1:10 chitin-acid) at 75 ^{° C for} 2 hours then added a hot distilled water (1: 5 chitin-water). And activated carbon (1: 0.5 chitin-carbon) and continued hydrolysis in a dilute aqueous acidic solution at ⁶⁰ ° C for another 2 hours. at the same time it cleans the hydrolyzate, and possibly also the catalytic effect of activated carbon. Hydrolysis at low temperature and acid concentration reduces the decomposition of D-glucosamine at this stage.

The resulting hot hydrolyzate was separated from coal by filtration on a Buchner funnel under vacuum. Coal was washed on the filter with hot water. The combined filtrate was evaporated in vacuo at a bath temperature not above ⁶⁰ ° C to reduce the liquid volume to 85% of the original volume. The final product was crystallized at a reduced temperature for 8 hours in an acid aqueous solution (not higher than 4 ° ^C). The precipitate of D-glucosamine hydrochloride was separated and washed with 96% alcohol to a white color. The precipitate was dried in a vacuum oven at a temperature not higher than 60 ^° C and alcohol regeneration of activated carbon causes no difficulties.

 The yield of the target product is 69% when the content of the main substance D-glucosamine hydrochloride is 99.4-99.6% by chloride ion and not less than 99% by the Elson-Morgan method.

PRI me R. 100 g of chitin, with an ash content not exceeding 0.3%, was loaded into a three-necked round-bottom flask with a capacity of 5 l, and 1000 g of conc. Hydrochloric acid (0.855 l) with a density of 1.17 (chemical grade) were poured. The mixture was heated for 2 hours at 75 ^{° C} and constant thorough mixing. Then to the flask were added 500 ml of hot (80 ° ^C) water was charged with 50 g of activated charcoal and heated with constant stirring for 2 hr at 60 ° ^C. By coal hydrolyzate was filtered hot on a Büchner funnel end of the hydrolysis and bleaching (cleaning) under vacuum (suction filter) through a three-four-layer filter. If the filtrate turned out to be contaminated with coal, it was heated to 60 ^° C and re-filtered. At the end of the filtration, the coal was washed on the filter with hot (70 ^° C) distilled water 500 ml (1: 5 chitin-water). Wash water was combined with the filtrate, which should be colorless or have a weak greenish-yellow color. If the filtered hydrolyzate has a dark color, then it is necessary to re-bleach it with charcoal (10: 1 chitin-charcoal) for 20 minutes at 60 ^o C.

Purified hydrolyzate was evaporated under vacuum (better in a rotary vacuum evaporator) at a pressure of 0.01 atm (1,013 kPa) and a water bath temperature of 60 ^{° C} with stirring to reduce the volume of the liquid 85%
Crystallization of D-glucosamine hydrochloride was performed from an acidic aqueous solution at a temperature not exceeding 4 ^{° C} for 8 hours under stirring.

 Upon completion of crystallization, the cooled suspension was filtered on a suction filter. The crystals were collected and transferred to a 500 ml beaker for washing with alcohol.

 100 ml of 96% ethanol was added to the crystals, stirred for 10-15 minutes and filtered to separate the alcohol. The washing was repeated three times in the same way as 100 ml, or 6 times in 50 ml (preferably 50 ml).

Washed the white crystals were dried in a vacuum oven at 55 ± ± 5 ^{° C} and a pressure of 0.01 atm (1,013 kPa) of moisture content of not more than 0.5% was obtained 69 g of D-glucosamine hydrochloride (69% yield). The filtrate was sent for alcohol regeneration, coal for washing and regeneration.

The desired product D-glucosamine hydrochloride is a white crystalline powder, odorless, hygroscopic, residual moisture 0.5% ash 0.2% Decomposition temperature (blackening test) 195-200 ^{° C,} pH of an aqueous 20% solution at 20 ^о С 2.3-2.5 units

Factors affecting the yield of D-glucosamine hydrochloride are:
1. Lower temperature, with an increase in the exposure of hydrolysis with the combination of hydrolysis and purification.

Time, h 2 2+ 2 2+ 3
Temperature, ^о С 75 75 and 60 75 and 60
Exit, 60 69 69
2. Dilution of the hydrolyzate with water, with continued hydrolysis in the presence of activated carbon.

Dilution
hydrolyzate
(chitin-water) 1: 1 1: 5 1:10
Concentration after
dilution if
accept conc.
that for 100% 91% 67% 50%
Exit, 63 69 65
3. Reduced evaporation temperature:
Temperature, ^о С 55 60 70
Exit 69 69 65
4. The crystallization temperature from an aqueous acidic solution:
Temperature, ^о С 4 10 15 20
Exit, 69 65 60 50
5. crystallization time at ⁴ ° C:
Time, h 7 8 9 10
Exit 69 69 69 69
6. The exclusion of the stage of precipitation of the product with alcohol and crystallization from an aqueous acidic solution allows to obtain large crystals, which reduces their surface and, accordingly, pollution. The precipitate is easily washed, filtered, does not clog the filter.

 7. The reduction in crystallization time compensates for the time spent on hydrolysis.

 Preliminary biological tests of D-glucosamine hydrochloride were carried out at the Institute of Rheumatology. Attached is a copy of the report-conclusion on the study of the chondroprotective (anti-arthritic) properties of D-glucosamine hydrochloride, performed by senior scientific researcher. Candidate of Medical Science N.V. Bunchuk and Junior Scientific al. V.V. Popov.

 D-glucosamine hydrochloride has a pronounced anti-arthritic effect, is part of the cartilage tissue of the joints.

 We conducted preclinical trials and obtained the permission of the Pharmacological Committee of the Ministry of Health of the USSR for conducting clinical trials in adults in the form of developed dosage forms (protocol of the Federal Ministry of Health of the USSR No. 9 of 05.16.1990).

 The acute toxicity of D-glucosamine hydrochloride when ingested exceeds 8000 mg / kg (800 g / 100 kg) of the weight of the animal, i.e. the substance is practically non-toxic. It is not inferior in effect to existing anti-arthritic drugs Rumalon and Arteparon, which are imported to the USSR in limited quantities and also have an allergenic effect, and D-glucosamine sulfate is produced abroad and does not enter the USSR.

 D-glucosamine hydrochloride significantly expands the arsenal of anti-arthritic drugs that are absent in the domestic medical industry.

 The inventive method allows to obtain an anti-arthritic agent D-glucosamine hydrochloride for medical use with a yield of 69% and a purity of 99% in comparison with the prototype (up to 64%).

 Upon receipt of D-glucosamine hydrochloride, the technological stage of converting it to base and sulfate is excluded, the stage of stabilization of sulfate by sodium chloride disappears, and the irritating effect of sodium chloride (13%) during intramuscular injection of the solution also decreases if stabilized glucosamine sulfate is taken. In addition, when ingesting glucosamine sulfate inside, it will go into chloride, because gastric juice has a strongly hydrochloric acid environment with a pH of about 1.8 units.

Claims (1)

METHOD FOR PRODUCING AN GLUCOSAMINE HYDROCHLORIDE HAVING ANTI-ARTHROSIS ACTIVITY, including grinding chitin, treating with hydrochloric acid while heating and stirring, then with water and activated carbon, evaporating in vacuo, crystallizing the target product, washing with ethanol, drying with 75 acid ^o With 2 hours, activated carbon and hot water with a ratio of chitin water of 1 5 at 60 ^o With for 2 hours, the evaporation of the hydrolyzate is also carried out at 60 ^o With, and the crystallization of the final product at 4 ^o C for 8 hours from the resulting hydrochloric acid solution.