CN101269215B - Glucoprotein incretion composition - Google Patents

Glucoprotein incretion composition Download PDF

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Publication number
CN101269215B
CN101269215B CN 200810037425 CN200810037425A CN101269215B CN 101269215 B CN101269215 B CN 101269215B CN 200810037425 CN200810037425 CN 200810037425 CN 200810037425 A CN200810037425 A CN 200810037425A CN 101269215 B CN101269215 B CN 101269215B
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composition
fsh
variant
hormone
glycoprotein
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CN 200810037425
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CN101269215A (en
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严惠敏
季斌
季晓铭
洪云海
高霄梁
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上海天伟生物制药有限公司
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    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
    • A61K9/00Medicinal preparations characterised by special physical form
    • A61K9/14Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
    • A61K9/19Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles lyophilised, i.e. freeze-dried, solutions or dispersions
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
    • A61K38/00Medicinal preparations containing peptides
    • A61K38/16Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/22Hormones
    • A61K38/24Follicle-stimulating hormone [FSH]; Chorionic gonadotropins, e.g. HCG; Luteinising hormone [LH]; Thyroid-stimulating hormone [TSH]

Abstract

The invention discloses a solid compound containing disaccharide and glycoprotein hormones or the variant thereof, wherein, the content of the glycoprotein hormones or the variant is not lower than 10 microgrammes per milligramme of compound or not lower than 100 international units per milligramme of compound. The compound provided by the invention has fine stability.

Description

一种糖蛋白激素组合物 A glycoprotein hormone composition

技术领域 FIELD

[0001] 本发明涉及蛋白质纯化和生物医药领域。 [0001] The present invention relates to the field of protein purification and bio-medicine. 具体而言,本发明涉及糖蛋白激素及其稳定剂的组合物,它具有良好的稳定性。 In particular, the present invention relates to glycoprotein hormones and stabilizer compositions having good stability.

背景技术 Background technique

[0002] 卵泡刺激素(Follicle-stimulating hormone,简称FSH)是由垂体产生的糖蛋白激素,它由α链和β链两个亚基组成。 [0002] FSH (Follicle-stimulating hormone, referred to as FSH) is produced by the pituitary glycoprotein hormones, which consists of two α chains and β chain subunits. FSH的α亚基与黄体生成激素(leuteinizing hormone,简称LH)和绒毛膜促性腺素(chorionicgonadotropin,简称CG)的α亚基完全相同,具有92个氨基酸,分子量约为14500D,第52和78位置上的天冬酰胺是发生N-糖基化的氨基酸。 Α subunit of FSH and luteinising hormone (leuteinizing hormone, referred to as LH) and chorionic gonadotropin (chorionicgonadotropin, referred to as CG) of the α subunit of identical 92 amino acid having a molecular weight of about 14500D, positions 52 and 78 asparagine is N- glycosylated amino acid occurs. FSH的β亚基由111个氨基酸组成,分子量约为18000D,其中第7和24位置上的天冬酰胺是发生N-糖基化的氨基酸。 Β subunit of FSH consisting of 111 amino acids, molecular weight of about 18000D, wherein the asparagine at position 7 and 24 is N- glycosylated amino acid occurs. 而LH的β亚基由121个氨基酸组成,分子量约为14800D ;CG的β亚基则有145个氨基酸,分子量22000-39000D。 While the LH β-subunit consists of 121 amino acids, molecular weight of about 14800D; CG of the β-subunit has 145 amino acids, molecular 22000-39000D.

[0003] 临床上FSH主要用于治疗不育症以及体外的辅助生殖。 [0003] FSH Clinically for the treatment of infertility and assisted reproduction in vitro. FSH可以从垂体或绝经期妇女的尿液中提取出来,也可通过DNA重组技术而制备。 FSH is obtained from urine, pituitary or in postmenopausal women, it may also be prepared by recombinant DNA techniques.

[0004] 含有FSH的第一代产品是尿促性素(HMG),如Serono公司的Pergonal,它是FSH与LH比例约为1的混合物。 [0004] FSH containing the first generation are hMG (HMG), Serono's as Pergonal, which is a mixture of FSH and LH ratio of approximately 1. 但是,对于体内有较多量的LH而不需加用外源性LH的患者,LH 水平过高会影响卵泡的正常发育,不合时机的抑制减数分裂抑制因子会导致卵子的老化, 从而降低受精和着床的机会。 However, the body has a greater amount of LH in patients without the addition of exogenous LH, LH levels are too high will affect the normal development of follicles, inhibiting untimely meiosis inhibitors can cause aging of the egg, thereby reducing fertilization and the chance of implantation. 因此对这部分患者而言,更适合于使用纯FSH制剂。 For some patients this thus, more suitable for use pure FSH preparations. 另一方面,过多的LH容易导致多囊卵巢综合症(Polycystic Ovarian Syndrome,P00S),研究表明, LH过多对生殖功能有不利的作用,如引起月经稀发、无排卵、不孕及流产。 On the other hand, too much LH easily lead to polycystic ovary syndrome (Polycystic Ovarian Syndrome, P00S), studies show, LH excessive adverse effects on reproductive function, such as those caused oligomenorrhea, anovulation, infertility and miscarriage . 因此对P00S患者用纯FSH治疗比HMG更为安全,可减少卵巢过度刺激综合症(OvarianHyperstimulation Syndrome, 0HSS)危险。 Therefore P00S patients with pure FSH treatment is more secure than HMG, may reduce ovarian hyperstimulation syndrome (OvarianHyperstimulation Syndrome, 0HSS) dangerous.

[0005] Serono公司推出的Metrodin是一种含有极少量LH的FSH制剂,其后的Metrodin-HP则是高纯度的FSH制剂。 [0005] Serono Metrodin has introduced a formulation containing a very small amount of FSH and LH, subsequent Metrodin-HP is a high purity FSH preparations.

[0006] 另外,近年来市场上也开发出一种高纯度的尿促性素 [0006] Further, in recent years, the market has developed a high purity hMG

Menopur,它是将纯化后的高纯度FSH与高纯度LH或者是高纯度hCG (人绒毛膜促性腺激素,human chorionic gonadotropin)按1 : 1的比例调配而成的产品,主要用于替代普通的HMG产品,并克服普通HMG产品中因大量杂蛋白而造成的对人体的过敏反应。 Menopur, which is a high purity of the purified high purity FSH LH or of hCG (human chorionic gonadotropin, human chorionic gonadotropin) and high purity by 1: 1 ratio of the deployment of products, mainly used to replace conventional HMG products, and overcome the body's allergic reaction to the product due to a large number of ordinary HMG contaminating proteins caused.

[0007] 由此可见,目前市场上都在致力于高纯度糖蛋白激素的开发和应用,但是,通过各种纯化手段获得的高纯度糖蛋白首先要做到一个原料药的形式,便于保存和检验,然后再做成制剂。 [0007] Thus, currently on the market are committed to the development and application of high-purity glycoprotein hormones, however, obtained through a variety of high-purity glycoprotein purification means the first to do a form of API, easy to save and test, and then made into formulations. 这个原料药的形式很重要,一般应尽量采用固体形式而不采用液体形式,因为液体不容易保存和运输,而对于糖蛋白激素来说固体形式的获得主要是通过冷冻干燥的手段,但我们知道,高纯度的糖蛋白激素在冷冻干燥的过程中很容易发生变性失活,因此现有技术中不得不采用液体的方式来保存高纯度糖蛋白激素的原料药,但液体形式必须保存在-20°C以下,否则容易失活;液体形式由于会遇到冻结_溶解过程,甚至是反复的冻结-溶解过程,更容易导致糖蛋白失活;液体形式还会遇到包装容器在低温下接近脆性点容易破裂的危险等。 This form of API is very important, the general should be used in solid form rather than using a liquid form because liquid is not easy storage and transport, and for the glycoprotein hormones is in solid form obtained mainly by means of freeze-drying, but we know , glycoprotein hormones with high purity in a freeze-drying process is prone to denaturation inactivated, thus the prior art have employed a manner to hold the liquid drug is a glycoprotein hormone of high purity, but it must be kept in liquid form -20 ° C or less, or easily inactivated; liquid form due to the encounter freeze _ dissolution process, and even repeated freezing - dissolution process more easily lead to inactivation of the glycoprotein; liquid packaging containers will encounter close brittle at low temperatures and so easy to spot the danger of rupture.

[0008] 因此本领域迫切需要开发出一种具有良好稳定性的糖蛋白激素的原料药形式。 [0008] Thus there is an urgent need to develop a drug having a good stability of form of the glycoprotein hormones. 发明内容 SUMMARY

[0009] 本发明旨在提供一种可作为原料药的含有高纯度的糖蛋白激素的固体组合物。 [0009] The present invention aims to provide high purity comprising a glycoprotein hormone a solid composition which may be used as a drug.

[0010] 在本发明的第一方面,提供了一种组合物,所述的组合物是固体组合物,其中含有二糖和糖蛋白激素或其变体。 [0010] In a first aspect of the present invention, there is provided a composition, said composition is a solid composition, which contains a disaccharide and a glycoprotein hormone or a variant thereof.

[0011] 在另一优选例中,所述的糖蛋白激素或其变体的含量为不低于10yg/mg组合物、 或不低于100国际单位/mg组合物。 [0011] In another preferred embodiment, the content of the glycoprotein hormones or a variant thereof is not less than 10yg / mg composition, or less than 100 IU / mg composition.

[0012] 在另一优选例中,所述的糖蛋白激素或其变体的含量为不低于20 μ g/mg组合物、 或不低于200国际单位/mg组合物。 [0012] In another preferred embodiment, the content of the glycoprotein hormones or a variant thereof is not less than 20 μ g / mg composition, or not less than 200 IU / mg composition.

[0013] 在另一优选例中,所述的糖蛋白激素或其变体选自卵泡刺激素(FSH)或其变体、 黄体生成素(LH)或其变体、或其混合。 [0013] In another preferred embodiment, the glycoprotein hormones or a variant thereof selected from follicle stimulating hormone (FSH) or a variant thereof, luteinizing hormone (LH) or a variant thereof, or mixtures thereof.

[0014] 在另一优选例中,所述的二糖选自乳糖、蔗糖、麦芽糖、海藻糖或其两种或两种以上的混合;更优选乳糖。 [0014] In another preferred embodiment, said disaccharide is selected from lactose, sucrose, maltose, trehalose, or a mixture of two or more than two; more preferably lactose.

[0015] 在另一优选例中,所述的卵泡刺激素或其变体是人卵泡刺激素或其变体。 [0015] In another preferred embodiment, the FSH or a variant thereof is human follicle-stimulating hormone or a variant thereof.

[0016] 在另一优选例中,所述的人卵泡刺激素或其变体选自重组的人卵泡刺激素或其变体、或人尿来源的卵泡刺激素或其变体。 [0016] In another preferred embodiment, the human FSH or a variant thereof selected from recombinant human FSH or a variant thereof, or human urine-derived follicle stimulating hormone or a variant thereof.

[0017] 在另一优选例中,所述的黄体生成素或其变体是人黄体生成素或其变体。 [0017] In another preferred embodiment, the luteinizing hormone or a variant thereof is human luteinising hormone or a variant thereof.

[0018] 在另一优选例中,所述的人黄体生成素或其变体选自重组的人黄体生成素或其变体、或人尿来源的黄体生成素或其变体。 Al. [0018] In another preferred embodiment, the human luteinizing hormone or a variant thereof selected recombinant luteinising hormone or a variant thereof, or human urine-derived luteinizing hormone or a variant thereof.

[0019] 在另一优选例中,所述的固体组合物是冻干粉。 [0019] In another preferred embodiment, the solid composition is a lyophilized powder.

[0020] 在另一优选例中,所述的组合物是固体,由二糖和糖蛋白激素或其变体组成,其中糖蛋白激素或其变体的含量为不低于10 μ g/mg组合物、或不低于100国际单位/mg组合物。 [0020] In another preferred embodiment, the composition is a solid, a disaccharide and a glycoprotein hormone or a variant thereof, wherein the content of a glycoprotein hormone or a variant thereof is not less than 10 μ g / mg composition, or less than 100 IU / mg composition.

[0021] 在本发明的第二方面,提供了一种药物组合物,所述的药物组合物含有如上所述的固体组合物。 [0021] In a second aspect of the present invention, there is provided a pharmaceutical composition, said pharmaceutical composition comprising a solid composition as described above.

[0022] 在本发明的第三方面,提供了一种如上所述的固体组合物在制备治疗不育综合症的药物中的用途。 [0022] In a third aspect of the present invention, there is provided the use of a solid composition as described above for the preparation of sterile syndrome therapeutic medicament.

[0023] 据此,本发明提供了一种具有良好稳定性的糖蛋白激素的原料药形式。 [0023] Accordingly, the present invention provides a drug having a good form stability of the glycoprotein hormones. 具体实施方式 Detailed ways

[0024] 发明人经过广泛而深入的研究,惊奇地发现高纯度的糖蛋白激素与合适的稳定剂组合进行冷冻干燥后,可以得到稳定的组合物。 [0024] The inventors of extensive and intensive studies, found that surprisingly high purity glycoprotein hormones suitable stabilizing agents in combination with freeze-dried to obtain a stable composition. 其中的稳定剂以二糖为佳,尤其是乳糖。 Wherein the stabilizer is preferably a disaccharide, especially lactose.

[0025] 发明人发现,此时二糖不仅起稀释剂和赋形剂的作用,更重要的是起保护剂的作用,它可以保护糖蛋白分子在冷冻干燥过程中可能发生的构象变化所导致的变性失活,另外一方面,它对产品的稳定性也有很大的提高。 [0025] The inventors have found that, at this time not only as a diluent and disaccharide excipients role, more important is the role of the protective agent that protects glycoproteins conformational changes may occur in the resulting freeze-drying process denatured inactivation, on the other hand, the stability of its products have greatly improved. 在此基础上,发明人完成了本发明。 On this basis, the inventors have completed the present invention.

[0026] 具体而言,如背景技术部分中所述,现有技术中采用了液体形式来保存高纯度糖蛋白激素的中间品或原料药形式,不便于保存和运输,稍有不慎,便会对产品产生很大的影响。 [0026] Specifically, as described in the background section, the prior art employed to store intermediate products in liquid form or in the form of highly pure drug glycoprotein hormones, inconvenient storage and transport, the slightest mistake, then the product will have a huge impact. 而采用固体形式又遇到冷冻干燥过程容易导致糖蛋白失活的结果。 The use of solid forms encountered during freeze-drying easily lead to inactivation of the glycoprotein results.

[0027] 在探索改进现有技术方法的过程中,发明人注意到将高纯度糖蛋白与乳糖进行适当的配比,特别是高纯度糖蛋白和乳糖的浓度在一个合适范围内,那么此时乳糖会起到保护剂的作用,防止糖蛋白分子在冷冻干燥过程中可能发生的构象变化所导致的变性失活。 [0027] In the process of the prior art methods of improving exploration, the inventors noted that high purity glycoprotein appropriate ratio with lactose, high purity particularly at a concentration of glycoproteins and lactose in a suitable range, then the time lactose will play the role of a protective agent, to prevent denaturation conformational change may occur glycoproteins in a freeze-drying process resulting from inactivation.

[0028] 在本发明中,所述的二糖(disaccharide)由二分子的单糖通过糖苷键形成,如麦芽糖、乳糖、蔗糖、海藻糖等;优选自下述的一种或多种,如麦芽糖、乳糖、和蔗糖;更佳地是乳糖。 [0028] In the present invention, the disaccharide (disaccharide) is formed from two molecules of monosaccharides through glycoside bonds, such as maltose, lactose, sucrose, trehalose and the like; preferably one or more from the following such as maltose, lactose, and sucrose; more preferably lactose.

[0029] 在本发明中,所述的糖蛋白激素的变体是指氨基酸发生缺失或通过重组技术对其进行修饰过的糖蛋白激素,但其基本药效还是存在或是得到增强。 [0029] In the present invention, the glycoprotein hormone variant refers to an amino acid deletion or modified glycoprotein hormone produced by recombinant techniques thereof, but the basic or enhanced efficacy still exist.

[0030] 本发明提供的组合物中,二糖和糖蛋白激素或其变体的重量之和是组合物总重量的60-100 %,较佳地是80-99 %,更佳地是90-95 %。 [0030] The compositions of the present invention is provided, disaccharides and glycoprotein hormones or a variant thereof and 60-100% by weight of the total weight of the composition, preferably a 80 to 99%, more preferably 90 -95%.

[0031] 在本发明的组合物中,所述的糖蛋白激素或其变体的生物效价为500-30000国际单位/mg蛋白质,较佳地为3500-12000国际单位/mg蛋白质。 [0031] In the compositions of the present invention, the glycoprotein hormones or a variant of the biological potency of 500-30000 IU / mg protein, preferably of 3500-12000 IU / mg protein. 如果其中的糖蛋白激素仅为FSH或其变体,FSH或其变体的生物效价为500-15000国际单位/mg蛋白质,较佳地为7000-12000国际单位/mg蛋白质;如果其中的糖蛋白激素仅为LH或其变体,LH或其变体的生物效价为500-8000国际单位/mg蛋白质,较佳地为2000-6000国际单位/mg蛋白质。 If only one of the glycoprotein hormones FSH or a variant thereof, or a FSH variant thereof biological potency of 500-15000 IU / mg protein, preferably of 7000-12000 IU / mg protein; wherein if sugar a protein hormone or a variant thereof only LH, or LH variant biological potency 500-8000 IU / mg protein, preferably 2000-6000 IU / mg protein.

[0032] 在本发明的组合物中,所述的糖蛋白激素或其变体的纯度为50%以上,较佳地为90%以上。 [0032] In the compositions of the present invention, the purity of the glycoprotein hormones or a variant thereof is 50% or more, preferably 90% or more.

[0033] 在本发明提供的每毫克组合物中,糖蛋白激素或其变体的含量不低于100国际单位,较佳地不低于200国际单位。 [0033] In the composition of the present invention to provide per mg, the content of a glycoprotein hormone or a variant thereof is not less than 100 international units, preferably not less than 200 IU.

[0034] 在本发明提供的每毫克组合物中,糖蛋白激素或其变体的含量不低于10 μ g。 [0034] In the composition of the present invention to provide per mg, the content of a glycoprotein hormone or a variant thereof is not less than 10 μ g. 如果其中的糖蛋白激素仅为FSH或其变体,FSH或其变体含量为不低于10μ g/mg组合物,较佳地不低于20 μ g/mg组合物;如果其中的糖蛋白激素仅为LH或其变体,LH或其变体的含量为不低于10 μ g/mg组合物,较佳地不低于35 μ g/mg组合物。 If only one of the glycoprotein hormones FSH or a variant thereof, or a FSH variant content is not less than 10μ g / mg composition, preferably not less than 20 μ g / mg composition; wherein if glycoprotein only LH hormone or variant thereof, the content of LH or a variant thereof is not less than 10 μ g / mg composition, preferably not less than 35 μ g / mg composition.

[0035] 本发明提供的组合物中,二糖和糖蛋白激素或其变体的重量比为8-99 : 1,较佳地为12-60 : 1。 [0035] The compositions of the present invention, provided, by weight of disaccharides and glycoprotein hormones or a variant ratio of 8-99: 1, preferably 12 to 60: 1.

[0036] 本发明提供的组合物中,乳糖和糖蛋白激素或其变体的重量比为8-99 : 1,较佳地为12-60 : 1。 [0036] The compositions of the present invention, provided, by weight of lactose and glycoprotein hormones or a variant ratio of 8-99: 1, preferably 12 to 60: 1. 如果其中的糖蛋白激素仅为FSH或其变体,乳糖和FSH或其变体的重量比为20-99 : 1,较佳地为30-50 : 1 ;如果其中的糖蛋白激素仅为LH或其变体,乳糖和LH 或其变体的重量比为8-99 : 1,较佳地为12-20 : 1。 If only one of the glycoprotein hormones FSH or a variant thereof, lactose or a FSH variant, and a weight ratio of 20-99: 1, preferably 30-50: 1; if only one of the glycoprotein hormones LH or a variant thereof, the weight ratio of lactose and LH or a variant thereof is 8-99: 1, preferably 12-20: 1.

[0037] 本发明提供的组合物在冷冻干燥过程中糖蛋白激素不会发生变性失活,并且此组合物具有良好的稳定性。 [0037] The present invention provides a composition does not denature the inactivation of the glycoprotein hormone in the freeze drying process, and the composition has good stability.

[0038] 本发明提供一种药物组合物,它包括:(a)糖蛋白激素或其变体;(b) 二糖;和(C) 药学上可接受的载体。 [0038] The present invention provides a pharmaceutical composition, comprising: (a) a glycoprotein hormone or a variant thereof; (b) a disaccharide; and the (C) a pharmaceutically acceptable carrier.

[0039] 如本文所用,术语“药学上可接受的载体”指用于治疗剂给药的载体,包括各种赋形剂和稀释剂。 [0039] As used herein, the term "pharmaceutically acceptable carrier" refers to a carrier for administration of therapeutic agents, including various excipients and diluents. 该术语指这样一些药剂载体:它们本身并不是必要的活性成分,且施用后没有过分的毒性。 This term refers to some of the medicament carrier: are not in themselves essential active components and without undue toxic after application. 合适的载体是本领域普通技术人员所熟知的。 Suitable carriers are those of ordinary skill in the art. 在Remington' s Pharmaceutical Sciences (Mack Pub. Co.,NJ 1991)中可找到关于药学上可接受的赋形剂的充分讨论。 In Remington 's Pharmaceutical Sciences (Mack Pub. Co., NJ 1991) can be found thorough discussion of pharmaceutically acceptable excipients. 在组合物中药学上可接受的载体可包括液体,如水、盐水、甘油和乙醇。 In the composition pharmaceutically acceptable carriers can include liquids such as water, saline, glycerol and ethanol. 另外,这些载体中还可能存在辅助性的物质,如崩解剂、润湿剂、乳化剂、PH缓冲物质等。 Additionally, these carriers may also exist in auxiliary substances, such as disintegrants, wetting agents, emulsifiers, PH buffering substances, and the like.

[0040] 所述药物组合物可以根据不同给药途径而制备成各种剂型。 The [0040] The pharmaceutical compositions can be prepared into various dosage forms according to different administration routes. 这些剂型以下面方式之一施用:口服、喷雾吸入、直肠用药、鼻腔用药、颊部用药、局部用药、非肠道用药,如皮下、 静脉、肌肉、腹膜内、鞘内、心室内、胸骨内和颅内注射或输入,或借助一种外植储器用药。 These dosage forms administered one of the following ways: orally, spray inhalation, rectal administration, nasal administration, buccal administration, topical administration, parenteral administration, such as subcutaneous, intravenous, intramuscular, intraperitoneal, intrathecal, intraventricular, intrasternal and intracranial injection or infusion, or by means of one of the explants medication reservoir. 其中治疗时优选皮下给药方式。 Wherein the treatment is preferably subcutaneous administration.

[0041] 另外需要指出,本发明药物组合物中有效成分的使用剂量和使用方法取决于诸多因素,包括患者的年龄、体重、性别、自然健康状况、营养状况、有效成分活性强度、服用、代谢速率、病症的严重程度以及诊治医师的主观判断。 [0041] Also to be noted, the pharmaceutical compositions of the invention in dosage and methods of use of the active ingredient depends on many factors, including the age, weight, gender, natural health status, nutritional status, active intensity of the active ingredient, taking, Metabolism the subjective judgment of the severity of the diagnosis and treatment as well as physicians rate condition. 建议安全有效量通常至少约10微克/ 千克体重,而且在大多数情况下不超过约10毫克/千克体重,较佳地该剂量是约10微克/ 千克体重-约1毫克/千克体重。 Recommended safe and effective amount typically at least about 10 micrograms / kg body weight, and in most cases not more than about 10 mg / kg body weight, preferably the dose is about 10 micrograms / kg body weight - about 1 mg / kg body weight. 当然,具体剂量还应考虑给药途径、病人健康状况等因素, 这些都是熟练医师技能范围之内的。 Of course, the specific dose factors should be considered the route of administration, patient health status, these are skilled physician within the skill range.

[0042] 本发明提到的上述特征,或实施例提到的特征可以任意组合。 [0042] The above features of the present invention mentioned above, or the features mentioned embodiments can be arbitrarily combined. 本案说明书所揭示的所有特征可与任何组合物形式并用,说明书中所揭示的各个特征,可以任何可提供相同、 均等或相似目的的替代性特征取代。 All of the features disclosed in this specification may be combined with and used in any combination thereof, various features disclosed in this specification may be any of the same may be provided, substituted alternative features equivalent or similar purpose. 因此除有特别说明,所揭示的特征仅为均等或相似特征的一般性例子。 Therefore, unless otherwise indicated, the disclosed feature is only an example of equivalent or similar features in general.

[0043] 本发明的主要优点在于: [0043] The main advantage of the present invention is that:

[0044] 1、提供了一种固体形式的糖蛋白激素的组合物,便于保存和运输; [0044] 1, provides a solid form of a composition glycoprotein hormones, to facilitate storage and transport;

[0045] 2、本发明方法获得的糖蛋白激素的组合物,具有良好的稳定性,作为原料药适用于制备相应的制剂。 [0045] 2. The method according to the present invention is obtained by a composition glycoprotein hormones, it has good stability, a respective formulations suitable for preparing a drug.

[0046] 下面结合具体实施例,进一步阐述本发明。 [0046] The following embodiments with reference to specific embodiments, further illustrate the present invention. 应理解,这些实施例仅用于说明本发明而不用于限制本发明的范围。 It should be understood that these embodiments are illustrative only and the present invention is not intended to limit the scope of the invention. 下列实施例中未注明具体条件的实验方法,通常按照常规条件、或按照制造厂商所建议的条件。 Experimental methods without specific conditions in the examples below, are performed under routine conditions, or in accordance with the conditions recommended by the manufacturer. 除非另外说明,否则百分比和份数按重量计算。 Unless otherwise indicated, percentages and parts are by weight.

[0047] 除非另行定义,文中所使用的所有专业与科学用语与本领域熟练人员所熟悉的意义相同。 [0047] Unless otherwise defined, all professional and scientific terms and those skilled in the text used in the familiar sense. 此外,任何与所记载内容相似或均等的方法及材料皆可应用于本发明中。 Further, any methods and materials similar or equivalent to those described herein Jieke applied to the present invention. 文中所述的较佳实施方法与材料仅作示范之用。 The method of the preferred embodiment described herein and the material for exemplary purposes only.

[0048] 实施例1 [0048] Example 1

[0049] 含FSH的组合物 [0049] FSH-containing compositions

[0050] 在本实施例中,将高纯度FSH(购自上海天伟生物制药有限公司)用作FSH药物组合物的原料,其中FSH的生物效价为9521国际单位/mg。 [0050] In the present embodiment, the high purity FSH (available from Shanghai Tian Wei Biological Pharmaceutical Co., Ltd.) was used as starting material in the pharmaceutical composition of FSH, wherein the FSH biological potency of 9521 IU / mg.

[0051] 配制50mL 0. OlM磷酸二氢钠溶液(用NaOH调pH约6. 5),加入2g乳糖,搅拌溶解后,用0.22μπι过滤器过滤。 [0051] formulated 50mL 0. OlM sodium dihydrogen phosphate solution (adjusted with NaOH to about pH 6.5), was added 2g of lactose and dissolved with stirring, filtered 0.22μπι filter. 取IOmL滤过液,加入10. Omg上述高纯度FSH(购自上海天伟生物制药有限公司),完全溶解后,装盘放入冻干机中,按如下程序进行冷冻干燥: Take IOmL filtrate was added 10. Omg said high purity FSH (available from Shanghai Tian Wei Biological Pharmaceutical Co., Ltd.). After complete dissolution, the loading into the lyophilizer and lyophilized according to the following procedure:

[0052] 1.层板至-40°C预冻3小时; [0052] 1. ply to pre-freezing -40 ° C 3 hours;

[0053] 2.冷阱至-45°C ; [0053] 2. A cold trap to -45 ° C;

[0054] 3.启动真空泵; [0054] 3. Start the vacuum pump;

[0055] 4.层板至-10°C维持15小时; [0055] 4. ply to -10 ° C for 15 hours;

[0056] 5.层板至+20°C维持5小时; [0056] The ply to + 20 ° C for 5 hours;

[0057] 6.出箱,得392mg冻干粉末。 [0057] 6 out of the box to give 392mg lyophilized powder.

[0058] 此产品测定FSH的生物效价结果如下:[0059] [0058] The results of the biological potency of FSH Related measured as follows: [0059]

Figure CN101269215BD00071

[0060] 对照例 [0060] comparative example

[0061] 在本对照例中,将高纯度FSH与乳糖配制成含FSH低于50国际单位/mg组合物总量的配比,再对其冷冻干燥过程及稳定性进行考察。 [0061] In the present Comparative Example, high purity FSH and FSH containing lactose formulated ratio lower than 50 IU / mg of the total composition, and then freeze-drying process and evaluate its stability.

[0062] 在本对照例中,将高纯度FSH(购自上海天伟生物制药有限公司)用作FSH药物组合物的原料,其中FSH的生物效价为9521国际单位/mg。 [0062] In the present Comparative Example, high purity FSH (available from Shanghai Tian Wei Biological Pharmaceutical Co., Ltd.) was used as starting material in the pharmaceutical composition of FSH, wherein the FSH biological potency of 9521 IU / mg.

[0063] 配制500mL 0. OlM磷酸二氢钠溶液(用NaOH调pH约6. 5),加入20g乳糖,搅拌溶解后,用0. 22 μ m过滤器过滤。 [0063] formulated 500mL 0. OlM sodium dihydrogen phosphate solution (adjusted with NaOH to about pH 6.5), 20g of lactose were added and dissolved with stirring, with a 0. 22 μ m filter. 取IOOmL滤过液,加入5. Omg上述高纯度FSH(购自上海天伟生物制药有限公司),完全溶解后,装盘放入冻干机中,按实施例1的程序进行冷冻干燥, 得3. 89g冻干粉末。 Take IOOmL filtrate was added 5. Omg said high purity FSH (available from Shanghai Tian Wei Biological Pharmaceutical Co., Ltd.). After complete dissolution, the loading into the lyophilizer and lyophilized according to the procedure of Example 1 to give 3. 89g lyophilized powder.

[0064] [0064]

Figure CN101269215BD00072

[0065] 从上述两例的结果可见,采用本发明所获得的FSH组合物,在冷冻干燥过程中几乎没有变性失活,而对照例有较明显的失活现象。 [0065] From the results of the above two examples, using FSH compositions of the present invention is obtained, in a freeze drying process is almost no denaturation inactivated, while the Comparative Example have a more significant deactivation.

[0066] 稳定性对比 [0066] Comparative Stability

[0067] 将实施例1和对照例的样品在30°C下考察其稳定性,结果如下 [0067] The sample of Example 1 and Comparative Example investigation at 30 ° C the stability, the following results

[0068] [0068]

Figure CN101269215BD00073

[0069] 可见,采用本发明所获得的FSH组合物与对照例相比具有良好的稳定性。 [0069] visible, FSH compositions obtained using the present invention compared with the comparative example having good stability.

[0070] 实施例2 [0070] Example 2

[0071] 含LH的组合物 [0071] LH-containing composition

[0072] 在本实施例中,将高纯度LH(购自上海天伟生物制药有限公司)用作LH药物组合物的原料,其中LH的生物效价为4307国际单位/mg。 [0072] In the present embodiment, the high purity LH (available from Shanghai Tian Wei Biological Pharmaceutical Co., Ltd.) was used as starting material in the pharmaceutical composition of LH, LH wherein the biological potency of 4307 IU / mg.

[0073] 配制50mL 0. OlM磷酸二氢钠溶液(用NaOH调pH约6. 5),加入2g乳糖,搅拌溶解后,用0. 22 μ m过滤器过滤。 [0073] formulated 50mL 0. OlM sodium dihydrogen phosphate solution (adjusted with NaOH to about pH 6.5), was added 2g of lactose and dissolved with stirring, with a 0. 22 μ m filter. 取IOmL滤过液,加入25mg上述高纯度LH(购自上海天伟生物制药有限公司),完全溶解后,装盘放入冻干机中,按实施例1的程序进行冷冻干燥,获得402mg冻干粉末。 Take IOmL filtrate was added 25mg of the high-purity LH (available from Shanghai Tian Wei Biological Pharmaceutical Co., Ltd.). After complete dissolution, the loading into the lyophilizer and lyophilized according to the procedure of Example 1, to obtain 402mg frozen a dry powder.

[0074] 此产品测定LH的生物效价以及在30°C下考察其稳定性,结果如下 [0074] Determination of Related LH biological potency and study its stability at 30 ° C, the following results

[0075] [0075]

Figure CN101269215BD00081

[0076] 可见,采用本发明所获得的LH组合物,几乎没有变性失活,且具有良好的稳定性。 [0076] visible, LH compositions obtained using the present invention, almost no denaturation inactivated, and has good stability.

[0077] 实施例3 [0077] Example 3

[0078] 含FSH、LH混合物的组合物 [0078] containing FSH, LH mixture composition

[0079] 配制IOOmL 0. 9%的氯化钠溶液(用NaOH或HCl调pH约7. 0),加入4g麦芽糖, 搅拌溶解后,用0.22μπι过滤器过滤。 [0079] formulated IOOmL 0. 9% sodium chloride solution (pH adjusted with NaOH or HCl to about 7.0), 4g of maltose was added, stirred to dissolve, filtered through a filter 0.22μπι. 取IOmL滤过液,加入10. Omg高纯度FSH(购自上海天伟生物制药有限公司,生物效价为9521国际单位/mg),再加入25. Omg高纯度LH (购自上海天伟生物制药有限公司,生物效价为4307国际单位/mg),完全溶解后,装盘放入冻干机中,按实施例1的程序进行冷冻干燥,获得405mg冻干粉末。 Take IOmL filtrate was added 10. Omg high purity FSH (available from Shanghai Tian Wei Pharmaceutical Company Ltd., the biological potency of 9521 IU / mg), was added high purity 25. Omg LH (available from Shanghai Tian Wei Biological pharmaceutical Co., Ltd., the biological potency of 4307 IU / mg), after complete dissolution, the loading into the lyophilizer and lyophilized according to the procedure of Example 1, to obtain 405mg lyophilized powder. 测得FSH生物效价为223IU/ mg, LH生物效价为242IU/mg。 Measured FSH biological potency of 223IU / mg, LH biological potency of 242IU / mg.

[0080] 此产品在30°C下的稳定性结果如下: [0080] Stability Results of this product at 30 ° C as follows:

[0081] [0081]

Figure CN101269215BD00082

[0082] 实施例4 [0082] Example 4

[0083] FSH冻干针剂的制备 Preparation of lyophilization of FSH [0083]

[0084] 用于制造100瓶FSH冻干针剂,且每瓶含75IU FSH的例子如下: Examples [0084] 100 for producing FSH lyophilization bottles and bottle containing 75IU FSH is as follows:

[0085] 将实施例1所制备的FSH冻干组合物35mg,溶解于IOmL注射用无热原水中,用0. 22 μ m过滤器进行无菌过滤,并用注射用无热原水洗涤过滤器。 [0085] FSH lyophilized composition prepared in Example 1 was 35mg, IOmL dissolved in pyrogen-free water for injection, sterile filtered with 0. 22 μ m filter, and washed with pyrogen-free water for injection was washed filter.

[0086] 将Ig乳糖溶解于30mL注射用无热原水中,如果需要的话,用HCl或NaOH调节PH6. 5士0. 2,用0. 22 μ m过滤器进行无菌过滤。 [0086] The lactose was dissolved in 30mL Ig pyrogen-free water for injection, if necessary, with HCl or NaOH to adjust PH6. 5 persons 0. 2, 0. 22 μ m sterile filtered with a filter. 然后加入到上述FSH溶液中,用注射用无热原水定容至75mL,混勻。 FSH was then added to the solution, with pyrogen-free water for injection to 75 mL volume, and mix.

[0087] 将上述溶液分装入安瓿瓶中,每瓶0. 75mL,进行冷冻干燥。 [0087] The above solution was dispensed into ampoules, bottle 0. 75mL, freeze-dried.

[0088] 所得到的安瓿瓶中,每瓶含约75IU FSH和10mg乳糖。 [0088] The obtained ampoules, each containing about 75IU FSH and 10mg of lactose.

[0089] 以上所述仅为本发明的较佳实施例而已,并非用以限定本发明的实质技术内容范围,本发明的实质技术内容是广义地定义于申请的权利要求范围中,任何他人完成的技术实体或方法,若是与申请的权利要求范围所定义的完全相同,也或是一种等效的变更,均将被视为涵盖于该权利要求范围之中。 [0089] The foregoing is only preferred embodiments of the present invention only, not intended to limit the scope of the substance of the technical contents of the present invention, the essence of the technical contents of the present invention is broadly defined in claim required range of application, any other person to complete technical or physical methods, exactly as if the application scope of the claims as defined, or an equivalent can change, will be regarded as encompassed within the scope of the claims.

Claims (4)

  1. 一种糖蛋白激素固体组合物,含有乳糖作为稳定剂;其中糖蛋白激素选自卵泡刺激素和黄体生成素,卵泡刺激素的生物效价为7000‑12000国际单位/mg蛋白质,黄体生成素的生物效价为2000‑6000国际单位/mg蛋白质;组合物中乳糖与卵泡刺激素的重量比为(30‑50)∶1;乳糖与黄体生成素的重量比为(12‑20)∶1。 A glycoprotein hormone solid composition containing lactose as stabilizing agent; wherein the glycoprotein hormone is selected from follicle stimulating hormone and luteinizing hormone, follicle stimulating hormone biological potency of 7000-12000 IU / mg protein, luteinizing hormone biological potency of 2000-6000 IU / mg protein; lactose composition with follicle stimulating hormone weight ratio of (30-50) to 1; luteinizing hormone lactose weight ratio of (12-20) to 1.
  2. 2.如权利要求1所述的固体组合物,其特征在于,所述的固体组合物是冻干粉。 Said solid composition as claimed in claim 1, wherein said solid composition is a lyophilized powder.
  3. 3.含有权利要求1-2中任一项所述固体组合物的药物组合物。 1-2 The pharmaceutical composition according to any one solid composition of claim 3 comprising.
  4. 4.权利要求1-2中任一项所述固体组合物在制备治疗不育综合症的药物中的用途。 The use of a solid composition in the manufacture of infertility syndrome therapeutic agent of any of claim 1-2.
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