SU833256A1 - Method of obtaining lactogenic hormone - Google Patents

Description

(54) METHOD FOR OBTAINING LACTOGENOUS HORMONE

; The invention relates to chemical-headlights of the manufacturing industry and relates to the acquisition of drugs that can be used in medical practice to stimulate lactadia in women born with hypogalactia.

A known method of producing lactogeic hormone is by dissolving acetonated loroshk from cattle hypophysis in acidified water, precipitating the hormone raw material with sodium chloride, dissolving the precipitate in water, dialysis the resulting solution against phosphate buffer, followed by dialysis against acetate buffer by centrifuging (1J.

However, the known method is long-lasting (12-14 days), labor-intensive (more than 30 operations) and provides an insufficiently high yield of lactogenic hormone (1500-3000 units / kg) with an activity 10 times lower than the outcome {| hormone.

The purpose of the invention is to increase activity, increase the yield of the target product and simplify the process.

This goal is achieved by the fact that a pathogenic hormone is obtained by dissolving n acetonated powder from cattle hypophysis in acidified water, precipitating the raw hormone sodium chloride, dissolving the precipitate in water. Dialysis of the resulting solution against phosphate buffer, followed by dialysis against. acetate buffer by centrifugation, the precipitate after salting out dissolved in water at pH 8.0-8.2, dialyzed against phosphate buffer pH 7.1-7.3, the supernatant filtered through kieselgur and then dialyzed. against acetate I-o buffer with a pH of 5.1-5.3.

The method is carried out as follows.

Precipitate (efflorescence) of a raw lactogenic hormone (LTG) is dissolved in water at pH 8.0-8.2, and the insoluble part is separated by centrifugation. The solution is placed in plastic bags and dialyzed against 0.4-0.7 molar phosphate buffer with a pH of 7.1-7.3 to. full, separated ballast substances (GO-40h). The precipitated precipitate is separated by centrifugation, and the supernatant is filtered through a kisel- layer. gur (0 5-2cm).

Claims (2)

The clear solution is placed in cellophane bags n dialdate against D, 05-0.2 molar acetate buffer with a pH of 5.1-5.3 for 18-24 hours. The precipitated precipitate is separated by centrifugation, dissolved in water, adjusted to pH to S and lofilizuyut. The output of the lactogenic hormone J5000-20000ЕД from 1 kg of the anterior lobes of the cat hypophysis. Activity hormone 15-20ED / MG. Electrophoresis in a full-acrylamide gel reveals 3 major bands characteristic of LTG. Example I. 50 g of acidic acetonating powder obtained from 4 kg of anterior cattle pituitary glands are taken, dissolved in 2 l of O, I acetic acid solution and 130 ml of saturated sodium chloride solution is added to the solution with stirring. The precipitate is allowed to form for 2 hours at 5 ° C, then centrifuged. The supernatant is used to produce corticotropin, and the precipitate is dissolved in water at pH 8.0. The resulting solution is poured into cellophane bags and dialyzed for 24 hours against a Yul 0.015 m phosphate buffer containing 0.5 mol of sodium chloride, pH 7.3. The precipitate formed during the dialysis process is separated by centrifugation. The supernatant is filtered through a layer of kieselguhr, another 2 cm. until a clear solution is obtained. The resulting solution is placed in plastic bags and dialyzed against 0.1 M sodium acetate buffer with a pH of 5.3 for 18 hours at 5 ° C. The precipitated LTG precipitated in the hfvtseee dialysis was separated by centrifugation, dissolved in water pM, and lofilized. Powder weight 3.2 g pr “a“; tivnostn 20.5 U / mg Example 2. Acting & amphibiously | y 1, the precipitate is dissolved in water at a pH of 8.2j and the resulting solution is placed in plastic bags and dialyzed against phosphate buffer at pH 7.1. The precipitate is separated and the supernatant is filtered through a 0.5 cm thick layer of kieselguhr. The filtration process is repeated until a clear solution is obtained. The resulting "s" in this way the solution is placed in cellophane bags and dialyzed against acetate buffer with a pH of 5.1 for 24 hours at 2 ° C. The formed LTG precipitate is separated by centrifugation, washed with acetone and dried in a vacuum oven at a temperature not higher than 45 ° C. Powder weight 5.5 g with an activity of 16.0 U / mg. Example 3. 100 g of acidic acetonated1-0 powder obtained from 5kg of the anterior parts of bovine pituitary glands are dissolved in 5 liters of distilled water and 2fi hydrochloric acid. The pH in pacTiiope is adjusted to 2.8-3.0. With stirring, 325 ml of solution is added to the solution: a solution of sodium chloride solution 11 is settled at a temperature of 2–8% and 3-4 hours. The precipitated precipitate is separated by centrifugation and redissolved in 2.5 liters of water, bringing the pH to 2.0-2.5. The insoluble portion is separated by centrifugation, and 160 ml of saturated sodium chloride solution is added to the supernatant. After 1-2 hours, the precipitate is collected by centrifugation, suspended in water and the pH is adjusted to 7.5-8.5. The volume of the resulting solution is 1 l. It is flooded with i double cellophane bags and dialyzed against 8 l of phosphate buffer with a pH of 6.4-6.8 for 36 hours. The precipitated after dialysis is separated by centrifugation and discarded, and the supernatant is placed in double cellophane bags n dialyzed against 8 l of acetate buffer with a pH of 5.4-5.8 for 36 hours. The precipitate which has precipitated after dialysis is separated by centrifugation, suspended in 500 ml of water, the pH value is adjusted to 3.0 with the addition of hydrochloric acid. 33 ml of a saturated sodium chloride solution are added thereto. The precipitated lactogenic hormone is re-dissolved in distilled water at a pH of 7.5-8.5, and the resulting solution is placed in plastic bags and dialnated against phosphate buffer for 24 hours. The siege is separated by centrifugation, and the supernatant is mixed into cellophane bags and dialyzed against acetate buffer for 24 hours. The resulting precipitate of lactogenic hormone is separated by centrifugation, washed with acetone n and dried until acetone is completely removed. The weight of dry powder of lactogenic hormone is 9.2 g, and its biological activity is 0.8 U / mg. The yield, from I kg of the anterior lobes of the cat hypophysis, is 1,472 U. The proposed method, in comparison with the known, allows to increase the specific activity of LTG from 0.5-1.0 U / mg to 15-20 U / mg and the quality of the odd solution excludes impurities of ballast proteins, which dramatically increases the effectiveness of the drug when its medical use, to increase the output from 1500-3000 U / kg to 15000-20000 U / kg, as well as to simplify the process by reducing the number of operations from 30 to 15, which shortens the process time by 3-4 times. The method of obtaining lactogenic hormone by dissolving acetonated powder from cattle hypophysis in acidified water, precipitating the raw hormone with sodium chloride, dissolving the precipitate in water, dialysis the resulting solution against phosphate buffer, followed by dialysis against acetate buffer and centrifuge. 5 8332566 peeling, which differs by SA teM, and then dialyzed against acetate in order to increase activity, increase the buffer with a pH of 5.1-5.3. Bjbt & oaa target product and simplify pro-sources of information, Cessa, sediment after diluted solution-taken into account in the examination  in water at a pH of 8D-8.2, the dial is PRO. I- Handbook for the production of organic matter buffer with a pH of 7.1-7.3 over-5 preparations. Ed. Turov V. A. and others. sedimentary water is filtered through Knseli., Medicine, 1970, p. 59.