Composition of glycoprotein scarcely containing subunits

A composition and glycoprotein technology, applied in the field of protein drug purification, can solve the problems of inactivation of glycoproteins, easy inactivation of glycoproteins, not easy to transport, etc.

Inactive Publication Date: 2012-01-11
SHANGHAI TECHWELL BIOPHARMACEUTICALS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Generally, solid form should be used as much as possible instead of liquid form. For APIs, liquid form is not easy to store and transport, because liquid form must be kept below -20°C, otherwise glycoprotein is easily inactivated; Freezing-thawing process, even repeated freezing-thawing process, is more likely to cause glycoprotein inactivation; liquid form also encounters the danger of easy rupture of packaging containers at low temperatures close to the brittle point, etc.
For the finished preparations, in addition to being difficult to transport in liquid form, the stability of glycoprotein drugs in liquid form is poor, and preservatives must be added to ensure that microorganisms will not grow within the validity period. For example, the publication number is Patent application of CN1309567A
[0008] The solid form is mainly obtained by means of freeze-drying, but high-purity glycoproteins are prone to denaturation and inactivation during the freeze-drying process. Subunits, and these subunits are also impurities, they are isomers that have no curative effect or may cause side effects, so the current freeze-dried preparations of high-purity glycoproteins must add 10-30% more amount when feeding to offset the freezing effect. The inactivation of the drying process, and a certain amount of subunits will be produced in the finished product, which reduces the purity of the product
The reasons for the degradation and inactivation are, on the one hand, the unreasonable formula of raw materials and auxiliary materials, and on the other hand, the freeze-drying process needs to be improved

Method used

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  • Composition of glycoprotein scarcely containing subunits
  • Composition of glycoprotein scarcely containing subunits
  • Composition of glycoprotein scarcely containing subunits

Examples

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preparation example Construction

[0062] The present invention also provides a method for preparing a glycoprotein composition, the method comprising the steps of:

[0063] (a) raising the temperature of the pre-frozen glycoprotein-containing aqueous solution to the sublimation drying temperature at a heating rate of 0.05-5° C. / minute under vacuum, and maintaining it for 1-30 hours; and

[0064] (b) Elevate the temperature to the analytical drying temperature at a rate of 0.05-5° C. / min, and maintain it for 1-20 hours to obtain a composition containing glycoprotein, and the composition preferably contains a freeze-dried powder of glycoprotein. Said composition is a composition of glycoproteins whose subunit content is not more than 10 wt%.

[0065] The advantages of the present invention are:

[0066] The freeze-drying method provided can reduce or hardly generate subunit degradation, thereby maintaining the activity of the glycoprotein, reducing the generation of impurities, and obtaining a glycoprotein almo...

Embodiment 1

[0078] Freeze-drying of FSH

[0079] Prepare 50mL of 0.01M sodium dihydrogen phosphate solution (adjust the pH to about 6.5 with NaOH), add 2g of lactose, stir to dissolve, and filter with a 0.22μm filter. Take 10 mL of the filtrate, add 10.0 mg of the above-mentioned high-purity FSH (purchased from Shanghai Tianwei Biopharmaceutical Co., Ltd.) (biological potency is 8817 international units / mg), after complete dissolution, the eutectic point of this solution is about -2 ℃, put it into a lyophilizer (purchased from Virtis), and freeze-dry according to the following procedure:

[0080] 1. Prefreeze the shelf at -40°C for 3 hours;

[0081] 2. Cold trap to -45°C;

[0082] 3. Start the vacuum pump;

[0083] 4. The shelf temperature is raised from -40°C to -10°C at a rate of 0.125°C / min, and the shelf temperature reaches -10°C and maintained for 15 hours;

[0084] 5. The shelf temperature is raised from -10°C to +20°C at a rate of 0.3°C / min, and the shelf temperature reaches +2...

Embodiment 2

[0101] Preparation of FSH freeze-dried injection

[0102] A typical example of the production of 10,000 vials of FSH freeze-dried injections, each containing 75IU FSH, is as follows:

[0103] A calculated amount (in units of biological potency) of FSH essence was dissolved in 50 mL of pyrogen-free water for injection, if necessary, adjusted to pH 6.5±0.2 with HCl or NaOH, and then sterile-filtered with a 0.22 μm filter .

[0104] Dissolve 100 g of lactose in 2 L of pyrogen-free water for injection, adjust the pH to 6.5 ± 0.2 with HCl or NaOH if necessary, and perform sterile filtration with a 0.22 μm filter. Then add it to the above FSH solution, dilute to 7.5L with pyrogen-free water for injection, and mix well. The eutectic point of this solution is about -2°C.

[0105] The above solution was divided into ampoules, 0.75mL per bottle, and freeze-dried according to the following procedure:

[0106] 1. Prefreeze the shelf at -40°C for 3 hours;

[0107] 2. Cold trap to -45°...

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Abstract

The invention discloses a composition of glycoprotein scarcely containing subunits. A method comprises the following steps of: (a) raising the shelf temperature of the pre-frozen aqueous solution containing glycoprotein to the range from 1 DEG C to 20 DEG C below the eutectic point of the solution at a temperature rate of 0.05-5 DEG C / min in vacuum, and maintaining the temperature for 1-30 hours;and (b) raising the shelf temperature to the temperature not lower than 0 DEG C at a temperature rate of 0.05-5 DEG C / min, and maintaining the temperature for 1-20 hours, thus obtaining lyophilized powder containing glycoprotein.

Description

[0001] This application is a divisional application of 200810042989.7, the filing date of the original application is September 17, 2008, and the title of the invention is "composition of glycoprotein with almost no subunits and its preparation method". technical field [0002] The invention relates to the field of purification of protein medicines. In particular, it relates to a freeze-drying method of an aqueous solution containing glycoprotein for treating infertility. Background technique [0003] Glycoproteins for the treatment of infertility are a class of substances with similar structures, including chorionic gonadotropin (HCG), menopausal gonadotropin (HMG), follicle stimulating hormone (FSH), and luteinizing hormone (LH). Phase gonadotropin is a mixture containing follicle-stimulating hormone and luteinizing hormone in a certain ratio (1:0.1-1). [0004] HCG, FSH, and LH are all composed of two subunits of α chain and β chain in the form of non-covalent bonds, in ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/24A61K9/19A61P15/08
Inventor 季晓铭高霄梁季斌严惠敏洪云海
Owner SHANGHAI TECHWELL BIOPHARMACEUTICALS CO LTD
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