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Slow-release pharmaceutical preparation of long-acting follicle-stimulating hormone

A technology of follicle-stimulating hormone and slow-release drugs, which is applied to medical preparations with non-active ingredients, medical preparations containing active ingredients, drug combinations, etc., and can solve problems such as high labor costs, affecting effects, and disturbing animals

Active Publication Date: 2016-06-29
HUAQIAO UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Various similar products currently require multiple injections, which not only has high labor costs, but also affects the effect due to multiple disturbing animals
Over the years, the industry has been researching on long-acting follicle-stimulating hormone, including genetic recombination, slow-release preparations, changing the route of administration, etc., in an attempt to reduce the number of injections, but so far no animal-use long-acting follicle-stimulating hormone has been launched on the market

Method used

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  • Slow-release pharmaceutical preparation of long-acting follicle-stimulating hormone
  • Slow-release pharmaceutical preparation of long-acting follicle-stimulating hormone
  • Slow-release pharmaceutical preparation of long-acting follicle-stimulating hormone

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Preparation of gene recombinant follicle-stimulating hormone:

[0028] The CHO cells with stable and high expression of FSH were inserted into shake flasks and gradually expanded to a density of 1×10 6 / ml, recorded as the first day of culture. After 14 consecutive days, the cell culture fluid was taken to purify the expression product.

[0029] The CHO cell culture medium was centrifuged at 9000 g at a low temperature of 4° C. for 20 min, and the culture supernatant was collected. The membrane ultrafiltration with a molecular weight cut-off of 10 kDa was used to collect the ultrafiltration concentrate. Purify the concentrated solution with ProteinA affinity chromatography column to obtain high-purity FSH protein with a purity of about 98% and a specific activity of 12000 IU / mg. Store at 2-8°C for later use.

Embodiment 2

[0031] Preparation of pituitary follicle-stimulating hormone long-acting injection:

[0032] Take 100,000 IU of pituitary follicle-stimulating hormone (purchased from Ningbo No. 2 Hormone Factory), and dissolve it in 200 ml of sodium citrate-citrate buffer solution (pH 6.8-7.2). Every 1 ml of sodium citrate-citric acid buffer (pH6.8-7.2) contains 30 mg of sodium citrate and 0.6 mg of citric acid. Take a low molecular weight heparin (molecular weight 5000) solution (200, or 400, or 600 ml) with a concentration of 10 mg / ml, and mix it with the pituitary follicle-stimulating hormone solution. Then take a protamine solution (600, or 400, or 200 ml) with a concentration of 10 mg / ml, and mix it with the above-mentioned solution of pituitary follicle-stimulating hormone and low molecular weight heparin. Add sodium citrate-citrate buffer to 1000ml. All preparation processes must be carried out in a class 100 sterile area. All solutions were sterile filtered before use. Under stirr...

Embodiment 3

[0037] In vitro dissolution test:

[0038] Take the preparations A1-3 obtained in Example 2, put them in 10ml centrifuge tubes, place them in a constant temperature incubator at 37°C, and centrifuge (3000rpm) at the 6th, 12th, 24th, 36th, 48th, 72nd, and 96th hours, and take the Clear 200 microliters, and measure FSH concentration by ELISA method. After each sampling, 200 μl of sodium citrate-citrate buffer (pH 6.8-7.2) was added to the centrifuge tube. According to the measurement results, the dissolution curve is obtained, such as figure 1 shown. Depend on figure 1 It can be seen that the dissolution rate of formulation A2 (the ratio of heparin to protamine is 1:1) is significantly slower than that of the other two, and is closer to the zero-order release rate.

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Abstract

The invention provides a slow-release pharmaceutical preparation of a long-acting follicle-stimulating hormone.The slow-release pharmaceutical preparation is prepared from the follicle-stimulating hormone, a sodium citrate-citric acid buffering solution, heparin and protamine.The follicle-stimulating hormone is a pituitary follicle-stimulating hormone or a gene recombinant follicle-stimulating hormone, the slow-release drug preparation is an injection or a freeze-dried injection, the ratio of the heparin to the protamine in the slow-release pharmaceutical preparation is 10:1 to 1:10, and 100-1000 IU of the follicle-stimulating hormone, 10-50 mg of sodium citrate, 0.1-1.0 mg of citric acid, 2-10 mg of heparin and 2-10 mg of protamine are added to every 1 ml of normal saline or water for injection.The slow-release pharmaceutical preparation can be applied to superovulation and estrus induction of animals.

Description

【Technical field】 [0001] The invention relates to a sustained-release pharmaceutical preparation with follicle-stimulating hormone as the main active ingredient, belonging to the fields of pharmacology, molecular medicine and disease prevention and treatment. 【Background technique】 [0002] Follicle-stimulating hormone (FSH) is a glycoprotein gonadotropin secreted by the anterior pituitary gland of animals. It has the functions of regulating the growth, sexual maturity and reproduction of the body. [0003] FSH can promote the growth of endometrium, ovulation, and stimulate the development of multiple follicles in female animals. There are specific receptors for FSH on the target cells. After FSH binds to the follicle-stimulating hormone receptor on the cell membrane, it activates adenylyl cyclase through the G protein coupling mechanism, increasing intracellular cAMP or causing calcium ion influx. , thereby affecting the metabolic or functional activities of cells. FSH b...

Claims

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Application Information

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IPC IPC(8): A61K38/24A61K47/36A61K47/42A61P15/08
CPCA61K9/0002A61K9/0019A61K9/08A61K9/19A61K38/24A61K47/36A61K47/42
Inventor 刁勇
Owner HUAQIAO UNIVERSITY
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