WO2012142961A1

WO2012142961A1 - Glycoprotein composition with low residual solvent level, and preparation method and use thereof

Info

Publication number: WO2012142961A1
Application number: PCT/CN2012/074425
Authority: WO
Inventors: 李勇; 季斌; 季晓铭
Original assignee: 上海天伟生物制药有限公司
Priority date: 2011-04-22
Filing date: 2012-04-20
Publication date: 2012-10-26
Also published as: KR20140018970A; CN102743742A

Abstract

Disclosed are a glycoprotein composition with low residual solvent level and preparation method thereof, the organic solvent content of the glycoprotein composition is no more than 0.5%; the method comprising the steps of: (a) vacuum drying the precipitate of a glycoprotein composition and an aqueous system together at 0-25°C; (b) continue vacuum drying after removing the aqueous system, and obtaining a glycoprotein composition with low residual solvent level.

Description

Glycoprotein composition with low solvent residue, preparation method and use thereof

The present invention relates to the field of purification of protein drugs. In particular, it relates to a method of drying a glycoprotein containing infertility. Background technique

Glycoproteins for the treatment of infertility are a class of structurally similar substances, including chorionic gonadotropin (HCG), menopausal gonadotropin (HMG), follic sputum hormone (FSH), luteinizing hormone (LH), in which menopause Gonadotropin is a mixture of follic sputum hormone and luteinizing hormone in a certain ratio (1: 0.1-1).

HCG, FSH, LH are all composed of two subunits, a chain and β chain, through non-covalent bonds, wherein their α subunits are identical, with 92 amino acids, molecular weight of about 14500D, positions 52 and 78. The asparagine is an amino acid that undergoes quinone-glycosylation. The β subunit of HCG has 145-147 amino acids with a molecular weight of 22200-39000D, and the asparagine at positions 13 and 30 and the positions 121, 127, 132, and 145 are where glycosylation occurs. The β subunit of FSH consists of 11 1 amino acids with a molecular weight of approximately 18000D, and asparagine at positions 7 and 24 is a Ν-glycosylated amino acid. The β subunit of LH consists of 121 amino acids with a molecular weight of approximately 14800D.

Clinically, HCG, HMG, FSH, and LH are mainly used to treat infertility and assisted reproduction in vitro. They can be extracted from the urine of a particular woman (pregnancy or menopause) or can be prepared by DNA recombination techniques.

The first generation of glycoproteins for infertility was Profono's Profas i (HCG) and Pergonal (HMG) in the 1960s, which were low in purity and contained large amounts of impurities. After the 1980s, Serono introduced Metrodi n-HP, a high-purity FSH preparation with low impurity content; later introduced Gonal-F (rFSH) and Luveri s (revolutionary technology) using DNA recombination technology. rLH), etc., Ferring also introduced high-purity menopausal gonadotropin-Menopur.

It can be seen that the development and application of high-purity glycoproteins are currently being pursued on the market to replace ordinary low-purity products to overcome the allergic reactions caused by the heteroproteins in ordinary low-purity products.

CN1309567A discloses liquid formulations containing FSH or variants thereof. However, because the liquid form must be kept below -20 °C, the glycoprotein is easily deactivated and the liquid form is not easy to store and transport; The liquid form is more likely to cause glycoprotein inactivation due to the freezing-dissolving process, or even the repeated freezing-dissolving process; the liquid form also encounters the risk of the packaging container being easily broken at a low temperature near the brittle point. For the finished product, the liquid form is not easy to transport, but also has poor stability of the liquid form of the glycoprotein drug, and must be added with a preservative to ensure that the microorganism will not breed during the effective period. A security hazard.

Therefore, the glycoprotein composition in solid form is more suitable for industrial production. The solid form is obtained mainly by freeze-drying and vacuum drying, but the high-purity glycoprotein is easily degenerated and deactivated during the freeze-drying process, and the denaturing inactivation mainly manifests in the complete molecular degradation into α chain and β. Two subunits of the chain, which are also impurities, which are isomers that have no curative effect or may cause side effects, and are added in an amount of 10-30% to offset the deactivation of the freeze-drying process, and A certain amount of subunits are also produced in the finished product, which reduces the purity of the product. CN101347613A discloses a lyophilization method for preparing a glycoprotein composition which can produce a high-purity, substantially subunit-free glycoprotein lyophilized composition, but the effect of removing the organic solvent by this method is not satisfactory.

Vacuum drying usually involves mixing the organic solvent with the glycoprotein solution, forming a precipitate and then dehydrating, and then drying the precipitate in a vacuum. This method can reduce the deactivation of glycoprotein, but the organic protein in the dried glycoprotein. The solvent content is high and the mass percentage exceeds 1%. Most of the organic solvents are harmful to the human body. In 2010, the Chinese Pharmacopoeia clearly stated that the ethanol limit in the drug was 0.5%.

Therefore, there is an urgent need in the art to develop a vacuum drying method capable of reducing the residual of an organic solvent in a glycoprotein, and thereby obtaining a glycoprotein composition having a low organic solvent residue. Summary of the invention

The present invention is directed to a glycoprotein composition having a low organic solvent residue, and a vacuum drying method for obtaining the above glycoprotein composition.

The 5%, preferably not more than 0.1%, more preferably not more than 0.1%, more preferably not more than 0.1%, more preferably not more than 0.1%.

Wherein the organic solvent is ethanol, acetone or methanol.

Wherein the glycoprotein is selected from the group consisting of chorionic gonadotropin, menopausal gonadotropin, follic sputum hormone, luteinizing hormone, or a mixture thereof.

Wherein the chorionic gonadotropin is a human urine source and/or a recombinant human chorionic gonadotropin, or a variant thereof; the menopausal gonadotropin is a human urine source and/or a recombinant human menopausal period a gonadotropin, or a variant thereof; the follicular sputum hormone is a human urinary source and/or recombinant human follicular stimulating hormone, or A variant thereof; the luteinizing hormone is human urine source and/or recombinant human luteinizing hormone, or a variant thereof. In another aspect of the present invention, there is provided a process for the simple preparation of a low solvent residual glycoprotein composition. The inventors have intensively studied and found that the glycoprotein composition and the water system are vacuum dried together, and the organic solvent can be effectively removed to obtain a low-solvent residual glycoprotein composition having good stability. Drying principle of composition of low solvent residual glycoprotein

Since glycoproteins are easily deactivated at high temperatures, it is not suitable to use spray drying or other heating methods for drying. The freeze-drying method also has a certain deactivation phenomenon, and the freeze-drying equipment is expensive, and the drying amount is small. It is expensive. The vacuum drying method (drying without water system) does not degrade the product, but the organic solvent remains high. The inventors conducted an in-depth analysis of the reasons for the excess solvent residue. The organic solvent and water volatilized together in a certain proportion in the early stage of vacuum drying of the sample, but the amount of volatilization increased with the decrease of the organic solvent and water content in the sample. Small, when the water content in the sample is no longer reduced, the amount of organic solvent is no longer reduced. Therefore, the residual amount of the organic solvent is related to the water content in the sample. The inventors have surprisingly found that placing a substance capable of releasing water vapor in a vacuum drier can not only obtain a composition of a glycoprotein having a low solvent residual, but also has no degradation inactivation of the sample.

The inventors conducted intensive studies on the drying principle after obtaining a composition of a low-solvent residual glycoprotein. A substance capable of releasing water vapor is placed in the vacuum dryer in order to control the ratio of the organic solvent to water in the sample during the vacuum drying process. During the vacuum drying process, the organic solvent volatilizes together with the water in the sample. In the vacuum drying oven, the organic solvent and the water vapor each occupy a certain proportion, but as the drying progresses, the moisture content is limited. In order to maintain this balance, we are In the drying system, a substance capable of releasing water vapor is added, and artificially increasing the water pressure in the vacuum drying box can suppress the volatilization of water in the sample, and at the same time promote the volatilization of the organic solvent in the sample. Therefore, the residual of the organic solvent in the sample is effectively reduced, and the water content in the sample is also maintained at a low level, so that the sample does not have degradation deactivation.

The inventors have further studied the drying process after obtaining the composition of the low-solvent residual glycoprotein, and surprisingly found that when the organic solvent is dried, the substance releasing the water vapor is removed, and the vacuum drying is continued for a certain period of time. The moisture in the sample can be vacuum dried to a qualified range, and the organic solvent can be further reduced, and the sample is stable in this moisture range. Therefore, the drying process of the present invention is divided into two stages, the first stage is to vacuum dry the sample and the water system together. The organic solvent, the second stage is to remove the water system and vacuum dry until the sample moisture content is qualified. Preparation

The present invention provides a method of preparing a glycoprotein composition having a low organic solvent content, the method comprising the steps of:

(a) The organic solvent is sufficiently mixed with a solution of the raw material of the glycoprotein-containing composition to precipitate, and the precipitate (1) is collected, and after dehydration with an organic solvent, the composition of the glycoprotein is further precipitated to obtain a precipitate containing the glycoprotein composition. (2) ; and

(b) The precipitate (2) containing the glycoprotein composition is vacuum dried together with the aqueous system to obtain a glycoprotein composition having a low organic solvent residue provided by the present invention.

In another preferred embodiment, in the step), the organic solvent has a temperature of -20 to 25 ° C, preferably - 20 to 0 ° C.

In another preferred embodiment, in the step (a), the organic solvent is selected from the group consisting of ethanol, acetone or methanol. In another preferred embodiment, in the step (a), the amount of the organic solvent for dehydration is from 0 to 10 times, preferably from 1 to 4 times the weight of the wet precipitate (1).

In another preferred embodiment, in the step (b), the drying temperature is from 0 to 25 ° C, preferably from 0 to 20 ° C.

In another preferred embodiment, in the step (b), the water system comprises solid, liquid or gaseous tap water, deionized water, reverse osmosis water, pure water and water for injection, an object capable of releasing water vapor, and an aqueous solution. Preferably, purified water is used.

In another preferred embodiment, in the step (b), the drying time is from 1 to 48 hours, preferably from 10 to 24 hours.

In another preferred embodiment, the solution containing the glycoprotein composition starting material described in step a can be prepared by the following steps:

The raw material containing the glycoprotein composition is dissolved in water or a buffer to prepare a solution containing the raw material of the glycoprotein composition.

In another preferred embodiment, the raw material containing the glycoprotein composition is a solid.

In another preferred embodiment, the buffer is selected from the group consisting of: buffers having a pH buffer ranging from 3 to 11; preferably from; phosphate buffer, Tris buffer, sodium acetate buffer.

In another preferred embodiment, the dissolution temperature is 5 to 20 ° C, preferably 10 to 16 ° C.

In another preferred embodiment, the solution containing the glycoprotein composition starting material has a pH of from 3 to 11, preferably from 4 to 10. In another preferred embodiment, the following steps may be further included after step (b):

(c) The vacuum system is removed after the water system is removed to obtain a composition of the low solvent residual glycoprotein provided by the present invention.

In the step (c), the drying time is from 0 to 48 hours, preferably from 5 to 24 hours. In a preferred embodiment of the invention, the method of preparing a low organic solvent content glycoprotein composition comprises the steps of:

(a') using an organic solvent of -20 to 0 ° C and a solution of the raw material of the composition containing the glycoprotein, and mixing and precipitating, and collecting the precipitate (1);

(b') the precipitate (1) is dehydrated with an organic solvent, and the composition of the glycoprotein is further precipitated to obtain a precipitate containing the glycoprotein composition (2);

(c') drying the composition precipitate of the glycoprotein (2) with a water system at 0 to 25 ° C for 1 to 48 hours to obtain a composition of a low solvent glycoprotein having good stability;

(d') After the step (c'), the water system is removed, and the low-solvent glycoprotein composition obtained in the vacuum drying step (c') is continuously dried at 0 to 25 ° C for 0 to 48 hours to obtain a lower solvent residue. a composition of glycoproteins. As used herein, "glycoprotein" is selected from one or more of the following: chorionic gonadotropin (CG), foll icule-stimulating hormone (FSH), luteotropic (luteotropic) Hormone, LH), human menopausal gonadotropin (HMG). As used herein, "glycoprotein composition material" means a composition containing a glycoprotein, including a glycoprotein, or a glycoprotein and a pharmaceutically/food acceptable carrier. The glycoprotein composition obtained by the method for preparing a glycoprotein composition existing in the art may be used as a raw material, and may be a solid or a liquid, such as, but not limited to, a high-purity HMG obtained by the preparation method disclosed in Chinese Patent No. CN1246332C; HCG obtained by the preparation method disclosed in the patent CN1958603B; and FSH obtained by the method disclosed in CN101317103A.

The above glycoprotein composition raw material is obtained by way of example only, and the glycoprotein composition raw material of the present invention should not be limited thereto.

In the method for obtaining a low solvent residual glycoprotein composition provided by the present invention, the "precipitating the precipitate containing the glycoprotein composition (2) together with the water system by vacuum" means that the sugar egg will be contained. The precipitate of the white composition (2) is placed in a place where the vacuum dryer is usually placed, and an open container containing a substance capable of releasing water vapor is placed around the vacuum dryer (for example, the bottom), which releases water The substance of the steam is selected from the group consisting of tap water, pure water, or a mixture of ice and water.

In the method of obtaining a low solvent residual glycoprotein composition provided by the present invention, the "withdrawn water system" means that the open container containing the substance capable of releasing water vapor is moved outside the vacuum drying environment. The main advantages of the invention are:

1. The present invention provides a novel low solvent residual glycoprotein composition.

2. The present invention provides a process for the preparation of a novel low solvent residual glycoprotein composition.

3. The invention has mild conditions, simple operation process, small degradation deactivation phenomenon, good product stability and low energy consumption, which greatly reduces the difficulty of process operation and reduces production cost. DRAWINGS

Figure 1 shows the GC spectrum of the standard with an ethanol content of 0.5%; wherein the retention time peak area and mass content of the solvent are listed in the following table:

Figure 2 shows the GC spectrum of ethanol in the HMG sample in Comparative Example 2;

Figure 3 shows the GC spectrum of ethanol in the HMG sample of Example 6;

Figure 4 shows a GC chromatogram of the ethanol content in the HCG sample of Example 9; Retention time (minutes) Peak area mass content (%) Solvent type

3.175 414.1 0.20 ethanol

detailed description

Determination of organic solvent residues in glycoprotein compositions

In an embodiment of the present invention, the residual solvent of the sample prepared by the method of the present invention is measured by GC, and the GC detection method is as follows:

Column: Varian CP-Select 624 (94% cyanopropylphenyl-6% dimethylpolysiloxane) Column size: 30mX0.32mmX 1.8μηι

Inlet temperature: 20CTC

Detector temperature: 26CTC

Flow rate: 2.5ml/min, constant current

Furnace temperature:

30 ° C / min

40 ° C, 2 ° C / min 5 ° C / min 250 ° C,

45 °C 80 °C clock

5 minutes 2 minutes Carrier gas: Nitrogen

Split ratio: 1 : 1 (workstation) , actual split ratio: 17:1 (measured by soap film meter) Detector gas: air flow: 400ml/min

Hydrogen flow: 40ml/min

Blowing gas flow: 25ml/min (nitrogen)

Headspace sampler conditions

Heating equilibrium temperature: 80 ° C; Quantitative ring temperature: 85 ° C; Transmission line temperature: 90 ° C; Equilibration time: 20 minutes; Pressurization time: 0.5 minutes; Filling time: 0.2 minutes; Quantitative ring equilibrium time: 0.10 minutes Injection time: 1 minute; Total running time: 30 minutes; Injection volume: 1ml; Shake mode: Slight vibration. The invention is further illustrated below in conjunction with specific embodiments. It is to be understood that the examples are merely illustrative of the invention and are not intended to limit the scope of the invention. The experimental methods in the following examples which do not specify the specific conditions are usually in accordance with conventional conditions or according to the conditions recommended by the manufacturer unless otherwise stated. Otherwise the percentage and number of copies are calculated by weight.

Unless otherwise defined, all professional and scientific terms used herein have the same meaning as those skilled in the art. In addition, any method-based material that is similar or equivalent to the recited content can be used in the method of the invention. The preferred embodiments and materials described herein are for illustrative purposes only.

The invention is further illustrated below in conjunction with specific embodiments. It is to be understood that the examples are not intended to limit the scope of the invention. The experimental methods in which the specific conditions are not indicated in the following examples are usually carried out according to conventional conditions or according to the conditions recommended by the manufacturer. Percentages and parts are by weight unless otherwise stated.

Unless otherwise defined, all professional and scientific terms used herein have the same meaning as those skilled in the art. In addition, any methods and materials similar or equivalent to those described can be applied to the present invention. The preferred embodiments and materials described herein are for illustrative purposes only. Comparative example 1

Freeze drying of FSH

45 mL of 0. 01M sodium dihydrogen phosphate solution (pH adjusted to about 6.5 with NaOH) was added, 5 ml of ethanol was added, 2 g of lactose was added, and the mixture was stirred and dissolved, and then filtered with a 0.22 μm filter. Take 10 mL of the filtrate, add 10. Omg of the above high-purity FSH (provided by Shanghai Tianwei Bio-Pharmaceutical Co., Ltd., the biopotency is 8817 IU/mg), and after completely dissolving, put the plate into the lyophilizer (purchased from Virt). In is), it was freeze-dried according to the method of Example 1 of CN101347613A, and was taken out of the box to obtain 391 mg of a lyophilized powder.

The biopotency results of FSH were determined as follows:

Comparative example 2

Preparation by traditional vacuum drying method

10 ml 0. lmol / L sodium acetate solution was prepared, adjusted to pH 6.5 with acetic acid, cooled to 10 ° C, and 250 mg of HMG raw material (provided by Shanghai Tianwei Bio-Pharmaceutical Co., Ltd., bioavailable by FSH 405 IU/mg) , stir to dissolve. Add 50 ml of -20 °C ethanol, stir and mix, centrifuge to obtain 931 mg of wet precipitate, dehydrate with 3 ml of ethanol, centrifuge to obtain 901 mg of HMG wet precipitate, and dry at 20 ° C under vacuum. At 96 hours, 249 mg of HMG sample was obtained. The bioavailability (in terms of FSH) and solvent content were determined as follows:

Example 3

Preparation of low solvent residues HMG

10 ml 0. lmol / L sodium acetate solution was prepared, adjusted to pH 4.0 with acetic acid, cooled to 16 ° C, added 250mg HMG raw material (provided by Shanghai Tianwei Bio-Pharmaceutical Co., Ltd., bio-valence 405 IU / mg in FSH) , stir to dissolve. 100 ml of ethanol at -20 ° C was added, stirred and mixed, and centrifuged to obtain 936 mg of HMG wet precipitate, which was vacuum dried at 20 ° C for 10 hours with an aqueous system containing an ice water mixture to obtain 257 mg of HMG sample. The bioavailability (in terms of FSH) and solvent content were determined as follows:

The results are as follows:

Example 4

Preparation of low solvent residues HMG

10 ml 0. lmol / L sodium acetate solution was prepared, adjusted to pH 6.5 with acetic acid, cooled to 10 ° C, and 250 mg of HMG raw material (provided by Shanghai Tianwei Bio-Pharmaceutical Co., Ltd., bioavailable by FSH 405 IU/mg) , stir to dissolve. Add 50 ml of -20 °C ethanol, stir and mix, centrifuge to obtain 956 mg of wet precipitate, dehydrate with 10 ml of ethanol, centrifuge to obtain 916 mg of HMG wet precipitate, and vacuum dry at 10 ° C for 24 hours with water system containing ice water mixture. 252 mg HMG sample. Determined, biologically effective The results are as follows:

Example 5

Preparation of low solvent residues HMG

Prepare 10ml 0. lmol/L sodium acetate solution, adjust pH10 with acetic acid, cool to 15 °C, add 250mg HMG raw material (provided by Shanghai Tianwei Bio-Pharmaceutical Co., Ltd., bio-valence FSIU 405IU/mg), stir and dissolve . Add 100 ml of ethanol at -10 ° C, stir and mix, centrifuge to obtain 953 mg of HMG wet precipitate, vacuum dry at 0 ° C with water system containing purified water for 24 hours, remove the water system, and continue drying at 0 ° C for 5 hours. 246 mg HMG sample. The bioavailability (in terms of FSH) and solvent content were determined as follows:

The results are as follows:

Example 6

Preparation of low solvent residues HMG

Prepare 10ml 0. lmol / L sodium dihydrogen phosphate solution, adjust pH 7.0 with NaOH solution, cool to 10 ° C, add 250mg HMG raw material (provided by Shanghai Tianwei Bio-Pharmaceutical Co., Ltd., bio-valence 405IU / FSH) Mg), stir to dissolve. Add 50 ml of ethanol at 0 ° C, stir and mix, centrifuge to obtain 952 mg of wet precipitate, dehydrate with 3 ml of ethanol, centrifuge to obtain 932 mg of HMG wet precipitate, and vacuum dry at 0 ° C with water system containing purified water for 24 hours to remove the water system. , continue to dry at 20 ° C for 24 hours. The 244 mg HMG sample was obtained, the biopotency (in terms of FSH) and the solvent content were as follows:

The results are as follows:

Example 7

Preparation of low solvent residues HCG

Prepare 10 ml of 0.1 mol/L Tris solution, adjust pH 7.5 with hydrochloric acid, cool to 10 ° C, add 300 mg of HCG raw material (provided by Shanghai Tianwei Bio-Pharmaceutical Co., Ltd., bio-potency 5702 IU/mg), stir and dissolve. Add 50 ml of ethanol at 0 ° C, stir and mix, centrifuge to obtain 1172 mg of wet precipitate, dehydrate with 11 ml of ethanol, centrifuge to obtain 1022 mg of wet precipitate of HCG, vacuum dry at 10 ° C with water system with ice water mixture for 24 hours, remove water System, continue to dry at 20 ° C for 24 hours, get

295 mg HCG sample.

The results are as follows:

Example 8

Preparation of low solvent residual FSH

Take 1000ml of solution containing FSH raw material at 10 °C (provided by Shanghai Tianwei Bio-Pharmaceutical Co., Ltd., bio-valence 1958 IU/ml), add 5000ml of ethanol at -20 °C, stir and mix, centrifuge to obtain 15820mg of wet precipitate, use 48ml Dehydration of ethanol, centrifuged to obtain 14958mg of FSH wet precipitate, vacuum drying at 0 °C with water system containing purified water for 24 hours, remove the water system, continue drying at 20 °C for 24 hours At the time, 3995 mg of FSH sample was obtained.

Example 9

Preparation of low solvent residual HCG

Take 10L of solution containing HCG raw material at 16 °C (provided by Shanghai Tianwei Bio-Pharmaceutical Co., Ltd., bio-valence 68319 IU/ml), add 50L of -10 °C ethanol, stir and mix, centrifuge to obtain 620g of wet precipitate, use 4L The ethanol was dehydrated, centrifuged to obtain 560 g of HCG wet precipitate, vacuum dried at 20 ° C for 20 hours with a water system containing purified water, the water system was removed, and drying was continued at 20 ° C for 20 hours to obtain 122 g of HCG sample.

The bioavailability and solvent content of HCG were determined as follows:

Example 10

Preparation of low solvent residues HCG

The procedure of Example 7 was followed except that the raw material was precipitated and dehydrated by replacing the ethanol in Example 7 to prepare a wet precipitate to obtain 298 mg of a HCG sample. The bioavailability and solvent content were determined as follows:

The results are as follows: Unit titer total potency titer yield methanol content water content

(IU/mg) (10,000 IU) (%) ( % ) ( % )

HCG raw materials 5702 171 1. 4 1. 5

HCG wet precipitation 59. 5 6. 2

HCG sample 5623 168 98 0. 08 1. 1

Example 11

Preparation of low solvent residues HMG

The procedure of Example 6 was followed except that acetone was used instead of the ethanol in Example 6 to precipitate and dehydrate the starting material to prepare a wet precipitate, thereby obtaining 240 mg of HMG sample. Determined, biological titer

FSH meter) and solvent content results are as follows:

The results are as follows:

The above is only the preferred embodiment of the present invention, and is not intended to limit the scope of the technical scope of the present invention. The technical content of the present invention is broadly defined in the scope of the claims of the application, any technical entity completed by others. The method or method, if it is identical to the scope of the claims, or equivalent changes, is considered to be within the scope of the claims.

Claims

Rights request

5%。 The content of the organic solvent is not more than 0.5%.

3%。 The organic solvent mass percentage does not exceed 0.3%.

1%。 The composition according to claim 2, wherein the organic solvent mass percentage does not exceed 0.1%.

The composition according to claim 1, wherein the organic solvent is ethanol, acetone or methanol.

The composition according to any one of claims 1 to 4, wherein the glycoprotein is selected from the group consisting of chorionic gonadotropin, menopausal gonadotropin, follic sputum hormone, luteinizing hormone, or a mixture thereof. .

The composition according to claim 5, wherein the human chorionic gonadotropin is human urine source and/or recombinant human chorionic gonadotropin, or a variant thereof; the menopausal gonadotropin Is a human urine source and/or recombinant human menopausal gonadotropin, or a variant thereof; the follicular sputum hormone is a human urinary source and/or recombinant human follicular sputum hormone, or a variant thereof; The human is a human urine source and/or recombinant human luteinizing hormone, or a variant thereof.

A method of preparing a glycoprotein composition according to any one of claims 1 to 6, wherein the method comprises the steps of:

(a) The organic solvent and the solution of the glycoprotein-containing composition raw material are thoroughly mixed and precipitated, and the precipitate (1) is collected, and after dehydration with an organic solvent, the composition of the glycoprotein composition is further precipitated to obtain a precipitate containing the glycoprotein composition. Object (2);

(b) The precipitate (2) containing the glycoprotein composition is vacuum dried together with the aqueous system to obtain the low solvent residual glycoprotein composition according to any one of claims 1 to 6.

The preparation method according to claim 7, wherein in the step a, the amount of the organic solvent for dehydration is 0 to 10 times, preferably 1 to 4 times the weight of the wet precipitate (1).

The preparation method according to claim 7, wherein in the step a, the organic solvent is selected from the group consisting of ethanol, acetone or methanol.

The preparation method according to claim 7, wherein in the step a, the organic solvent has a temperature of -20 to 25 ° C, preferably -20 to 0 ° C.

The preparation method according to claim 7, wherein in the step b, the vacuum drying temperature is 0 to 25 ° C, preferably 0 to 20 ° C.

The preparation method according to claim 7, wherein in the step b, the vacuum drying time is 1 to 48 hours, preferably 10 to 24 hours.

The preparation method according to claim 7, wherein in the step b, the water in the water system is tap water, deionized water, reverse osmosis water, pure water, or water for injection.

14. The method according to claim 13, wherein in step b, the water in the water system is in a liquid, solid, gaseous, or mixed form.

The preparation method according to claim 14, wherein in the step b, the water system is an aqueous solution.

The preparation method according to claim 15, wherein in the step b, the water system is an object that can generate water vapor.

The preparation method according to claim 7, wherein the solution containing the glycoprotein composition raw material described in the step a can be prepared by the following steps:

The production method according to claim 17, wherein the dissolution temperature is 5 to 20 ° C, preferably 10 to 16 ° C.

The preparation method according to claim 17, wherein the buffer is selected from the group consisting of: a buffer having a pH buffer ranging from 3 to 11; preferably from: phosphate buffer, Tris buffer, sodium acetate Buffer.

The production method according to claim 17, wherein the pH of the solution containing the glycoprotein composition raw material is controlled to be 3 to 11, preferably 4 to 10.

The preparation method according to claim 7, wherein the step b may further comprise the following steps:

(c) The vacuum system is removed after the water system is removed to obtain the low solvent residual glycoprotein composition of any of claims 1-6.

The preparation method according to claim 21, wherein in the step c, the vacuum drying time is 0 to 48 hours, preferably 5 to 24 hours.

23. Use of a glycoprotein composition according to any one of claims 1 to 6 wherein the composition is for the preparation of a medicament for the treatment of infertility syndrome.