WO2021208290A1 - Use of gphb5 glycoprotein hormone in lowering lipid and improving insulin sensitivity - Google Patents

Use of gphb5 glycoprotein hormone in lowering lipid and improving insulin sensitivity Download PDF

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WO2021208290A1
WO2021208290A1 PCT/CN2020/105383 CN2020105383W WO2021208290A1 WO 2021208290 A1 WO2021208290 A1 WO 2021208290A1 CN 2020105383 W CN2020105383 W CN 2020105383W WO 2021208290 A1 WO2021208290 A1 WO 2021208290A1
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gphb5
glycoprotein hormone
lipid
hormone
glycoprotein
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PCT/CN2020/105383
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French (fr)
Chinese (zh)
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赵子建
萧耿苗
李芳红
赵正刚
千爱君
张馨丹
王帅
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广州华真医药科技有限公司
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/22—Hormones
    • A61K38/24—Follicle-stimulating hormone [FSH]; Chorionic gonadotropins, e.g. HCG; Luteinising hormone [LH]; Thyroid-stimulating hormone [TSH]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/04—Anorexiants; Antiobesity agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/06—Antihyperlipidemics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
    • A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P5/00—Drugs for disorders of the endocrine system
    • A61P5/48—Drugs for disorders of the endocrine system of the pancreatic hormones
    • A61P5/50—Drugs for disorders of the endocrine system of the pancreatic hormones for increasing or potentiating the activity of insulin

Definitions

  • the present invention relates to the field of metabolic system diseases. Specifically, the present application provides the application of GPHB5 glycoprotein hormone in the preparation of drugs for reducing obesity and lipid-lowering, or drugs for treating glucose intolerance or insulin resistance in obese people; the present invention provides a A glycoprotein hormone GPHB5 with weight loss and fat reduction effects, reveals a new mechanism by which the pituitary gland regulates body weight and fat metabolism.
  • Obesity is a very common preventable cause of death and one of the most important public health problems in the 21st century. At present, the prevalence of obesity in both adults and children is on the rise, and it occurs more frequently in women than in men. From 1979 to 2016, the number of obese adult women increased from 69 million to 390 million, and obese men increased from 31 million to 281 million. Obesity will bring a series of complications, such as type 2 diabetes, cardiovascular disease, infertility, asthma, sleep apnea syndrome, etc. It is also related to the occurrence of breast cancer, rectal cancer, prostate cancer and other malignant tumors. It has seriously affected the health and quality of life of Chinese people.
  • GPHB5 is a subgroup of CGH, a member of the glycoprotein hormone family group, and is mainly secreted by ACTH-secreting cells in the anterior pituitary gland.
  • Glycoprotein hormone family members include follicle stimulating hormone (FSH), luteinizing hormone (LH), thyroid stimulating hormone (TSH), chorionic gonadotropin (CG), and CGH, a new glycoprotein hormone family member, was confirmed in 2002 .
  • FSH follicle stimulating hormone
  • LH luteinizing hormone
  • TSH thyroid stimulating hormone
  • CG chorionic gonadotropin
  • CGH a new glycoprotein hormone family member
  • GPHB5 can bind to TSH receptor (TSHR), and after GPHB5 and GPHA2 combine to form a heterodimer CGH, the ability to activate TSHR to produce cAMP is 4 times stronger than TSH.
  • activating adipose tissue TSHR can promote the hydrolysis of fat triglycerides, thereby reducing fat content and reducing weight. Therefore, GPHB5 is very likely to be combined with TSHR of adipose tissue to stimulate triglyceride hydrolysis, which can be used as a potential molecular target for weight loss and fat reduction.
  • the main problem that the present invention solves is to provide a new mechanism for weight loss and fat reduction, and treatment of glucose intolerance or insulin resistance in obese people; on the basis of existing research, the present invention proposes that the GPHB5 glycoprotein hormone can reduce fat in obesity. And the application in the treatment of glucose intolerance or insulin resistance in obese people; based on the role of GPHB5 glycoprotein hormone in weight loss and fat loss, it reveals a new mechanism for the pituitary to regulate body weight and fat metabolism.
  • the present invention relates to a glycoprotein hormone beta 5 (GPHB5) with the effect of reducing weight and fat.
  • GPHB5 glycoprotein hormone beta 5
  • the present invention provides the application of GPHB5 glycoprotein hormone in the preparation of drugs for reducing obesity and lipid-lowering.
  • the GPHB5 glycoprotein causes weight loss.
  • the GPHB5 glycoprotein reduces the body fat content, but the non-fat content does not change significantly.
  • the present invention provides the application of GPHB5 glycoprotein hormone in the preparation of drugs for the treatment of glucose intolerance, insulin resistance or diabetes in obese people.
  • the GPHB5 glycoprotein hormone can be GPHB5 glycoprotein hormone purified from tissues or recombinantly expressed.
  • the GPHB5 glycoprotein hormone can be prepared as an oral or injection dosage form.
  • the present invention provides an obesity-lowering drug, which is characterized in that it contains GPHB5 glycoprotein hormone, or a vector or cell expressing GPHB5 glycoprotein hormone.
  • the drug effect includes weight loss, or reduction in body fat content but no significant change in fat content.
  • the present invention provides a medicine for treating glucose intolerance or insulin resistance in obese people, which is characterized in that it contains GPHB5 glycoprotein hormone, or a vector or cell expressing GPHB5 glycoprotein hormone.
  • the medicine can be in the form of oral or injection.
  • the GPHB5 glycoprotein hormone in the present invention includes GPHB5 glycoprotein hormone from various biological sources.
  • the GPHB5 glycoprotein hormone in the present invention can be GPHB5 glycoprotein hormone purified from tissues using various known separation and purification methods or recombinantly expressed.
  • the GPHB5 glycoprotein hormone in the present invention does not exclude the existence of other amino acid sequences, as long as it does not hinder the function of the GPHB5 glycoprotein hormone.
  • the medicine of the present invention may contain various excipients, vectors, and expression cells that are known or unknown in the pharmaceutical field/biological field, and those skilled in the pharmaceutical field/biological field can design corresponding dosage forms based on existing knowledge.
  • the present invention provides a method for reducing obesity and lipid reduction, including administering GPHB5 glycoprotein hormone to a subject in need.
  • the present invention provides a method for treating glucose intolerance, insulin resistance or diabetes in obese people, including administering GPHB5 glycoprotein hormone to subjects in need.
  • the GPHB5 glycoprotein hormone is a GPHB5 glycoprotein hormone purified from tissues or recombinantly expressed.
  • the present invention has the following beneficial effects:
  • the present invention is based on the role of the GPHB5 glycoprotein hormone in weight loss and fat reduction, and reveals a new mechanism for the pituitary to regulate body weight and fat metabolism.
  • the invention can help lose weight, enhance insulin sensitivity, and improve blood lipid metabolism through injection or patients with low GPHB5 recombinant hormone levels.
  • Figure 1 shows the GPHB5 knockout verification. GPHB5 mRNA expression levels in the testis, brain, and heart of wild-type mice and knockout mice were detected by qPCR. GPHB5 mRNA in the three tissues of knockout mice was significantly lower than that of wild-type mice, and almost no expression. n.d. stands for not detectable.
  • Figure 2 shows the body weight growth curve of GPHB5 knockout mice (-/-) and littermate wild-type mice (WT).
  • Figure 3 is a comparison of the body types of GPHB5 knockout mice (-/-) and littermate wild-type mice (WT).
  • Figure 4 shows the comparison of fat content (Fat Mass) and non-fat content (Lean Mass) of GPHB5 knockout mice (-/-) and litter wild-type mice (WT).
  • FIG. 5 is a comparison of glucose tolerance (GTT) between GPHB5 knockout mice (-/-) and litter wild-type mice (WT).
  • Figure 6 is a comparison of insulin tolerance (ITT) between GPHB5 knockout mice (-/-) and litter wild-type mice (WT).
  • Figure 7 is a schematic diagram of the expression vector pBudCE4.1-GPHB5/GPHA2 plasmid.
  • Figure 8 is a schematic diagram of the expression vector pcDNA3.1-GPHB5-L-GPHA2 plasmid.
  • Figure 9 shows the Western Blot detection of recombinant GPHB5/GPHA2 protein.
  • Figure 10 shows the change of cAMP in cells after 3T3-L1 cells stimulated by recombinant protein by ELISA.
  • the lack of GPHB5 glycoprotein hormone will cause weight gain. Specifically, the lack of GPHB5 will increase the body fat content, while the non-fat content will not change significantly. The lack of GPHB5 will cause glucose intolerance and insulin resistance.
  • the application includes the following steps: using GPHB5 protein purified from tissues or recombinantly expressed and purified by injection or oral administration to reduce weight and lipids in obese patients; or using GPHB5 protein purified from tissues or recombinantly expressed and purified by injection Or it can be taken orally to improve insulin sensitivity and lower blood sugar in obese patients.
  • the Genbank number of the mouse GPHB5 gene is NM_175644.3, the Genbank number of the human GPHB5 gene is NM_145171.4, and the full length of the GPHB5 gene is about 3900bp.
  • the GPHB5 knockout mice used were commissioned by Saiye Biotechnology Co., Ltd., through the CRISPR/Cas9 gene knockout technology, the 2 and 3 exons of the GPHB5 gene were knocked out, and the 3340bp base was knocked out to obtain 2 GPHB5 knockout mice. Labeled as line1 and line2.
  • the first 10 sites that may be off-target were detected by DNA sequencing, and the results showed that line1 and line2 were not off-target, indicating that the GPHB5 knockout mouse was successfully constructed.
  • GPHB5 knockout mice and littermate wild-type mice were obtained.
  • the testes, brains, and hearts of GPHB5 knockout mice and wild-type mice were taken to detect GPHB5mRNA levels.
  • Figure 1 the results showed that GPHB5 knockout mice basically did not express GPHB5mRNA.
  • mice were reared in an SPF barrier environment. Male mice in the same litter were placed in the same cage, with 3-4 mice per cage, and fed with the same feed and sterile water. The body weight of the mice was recorded after 6 weeks of age and was recorded once a week, as shown in Figure 2 and Figure 3. After 18 weeks of age, the body weight of GPHB5 knockout mice was significantly higher than that of wild-type mice in the same litter. At 31 weeks, the body weight of GPHB5 knockout mice was 6g more than that of wild-type mice, indicating that GPHB5 deletion can lead to obesity.
  • the body composition analyzer Echo MRI of Huijia Biological Co., Ltd. was used to detect the body fat content and non-fat content of GPHB5 knockout mice and littermated wild-type mice. As shown in Figure 4, the body fat content of GPHB5 knockout mice was significantly higher than that of wild-type mice in the same litter, while the non-fat content was basically the same, indicating that the increase in body weight of GPHB5 knockout mice was mainly due to the accumulation of fat.
  • GPHB5 knockout mice and wild-type mice in the same litter were fasted for 12 hours, and the fasting blood glucose was measured by Roche blood glucose meter. According to 2g/kg body weight, 20% glucose was injected intraperitoneally, and the blood glucose level of mice 15min, 30min, 60min, 90min, 120min, 150min after injection was measured by Roche blood glucose meter, as shown in Figure 5, the results showed the glucose tolerance of GPHB5 knockout mice It is significantly lower than that of wild-type mice in the same litter.
  • the complete gene sequences of GPHB5 and GPHA2 were synthesized by Nanjing GenScript Company and optimized by humanization of codons.
  • the target gene was cloned into the pBudCE4.1 vector, where the GPHB5 gene was cloned between the Hind III/BamH I sites of the vector, and the GPHA2 gene was cloned between the Not I/Xho I sites of the vector, as shown in Figure 7.
  • the pBudCE4.1-GPHB5/GPHA2 plasmid was transformed into MJ109 Escherichia coli, and a single clone was picked for plasmid identification and DNA sequencing to verify the correctness of the inserted sequence.
  • pBudCE4.1 plasmid is a commonly used vector for expressing double genes in mammalian cells.
  • the vector carries His tag, V5 tag and myc tag sequence.
  • GPHB5 protein will carry myc and His tag after expressing recombinant protein, GPHA2
  • the protein will carry V5 and His tags, which can be used for purification and detection.
  • the fusion gene containing the signal peptide of GPHB5 and the mature peptide, the connecting peptide L, the mature peptide of GPHA2 and the His tag was synthesized, and cloned into the EcoR I/Not I site of the pCDNA3.1 vector, as shown in Figure 8.
  • connecting peptide L there are two protein sequences of connecting peptide L: the conventional connecting peptide GGGGSGGGGSGGGGS, and the CTP peptide PRFQDSSSSKAPPPSLPSPSRLPGPSDTPILPQ, which can extend the half-life of the recombinant protein.
  • the constructed expression vector pBudCE4.1-GPHB5/GPHA2 and the expression vector pcDNA3.1-GPHB5-L-GPHA2 were transfected into mammalian cells, including HEK 293T cells, 293F cells, and CHO cells.
  • Figure 9 shows the result of Western Blot experiment with the culture supernatant after transfection of the expression vector pBudCE4.1-GPHB5/GPHA2 into 293T cells, which indicates that the GPHB5/GPHA2 recombinant protein was successfully expressed.
  • the culture medium containing the recombinant protein was concentrated by ultrafiltration tube 5 times. After purification by the His tag purification kit, it was used to stimulate 3T3-L1 cells for 20 min, 60 min, and 90 min.
  • the intracellular cAMP level was detected by ELISA, and it was found to be compared with the control group.
  • the expressed GPHB5/GPHA2 recombinant protein stimulates cAMP to increase after 20min and 60min, as shown in Figure 10, indicating that the expressed recombinant protein is active.

Abstract

Provided in the present invention is use of GPHB5 glycoprotein hormone for preparing a medicament for reducing obesity and lipid or a medicament for treating glucose intolerance, insulin resistance or diabetes in obese populations. The use comprises using purified or recombinantly expressed and purified GPHB5 protein from a tissue to reduce weight and lipid in obese people or to improve insulin sensitivity and reduce blood sugar in obese people by means of injection or oral administration. The present invention reveals a new mechanism for regulating body weight and lipid metabolism by pituitary gland on the basis of the effect of GPHB5 glycoprotein hormone in weight loss and lipid reduction.

Description

GPHB5糖蛋白激素在降脂和改善胰岛素敏感中的应用Application of GPHB5 Glycoprotein Hormone in Lowering Lipid and Improving Insulin Sensitivity 技术领域Technical field
本发明涉及代谢系统疾病领域，具体地，本申请提供了GPHB5糖蛋白激素在制备减少肥胖以及降脂药物，或治疗肥胖人群葡萄糖不耐受或胰岛素抵抗的药物中的应用；本发明提供了一种具有减肥减脂功效的糖蛋白激素GPHB5，揭示一种垂体调控体重和脂肪代谢的新机制。The present invention relates to the field of metabolic system diseases. Specifically, the present application provides the application of GPHB5 glycoprotein hormone in the preparation of drugs for reducing obesity and lipid-lowering, or drugs for treating glucose intolerance or insulin resistance in obese people; the present invention provides a A glycoprotein hormone GPHB5 with weight loss and fat reduction effects, reveals a new mechanism by which the pituitary gland regulates body weight and fat metabolism.
背景技术Background technique
肥胖是一种很常见的可预防死因，也是21世纪最重要的公共卫生问题之一。目前成人与儿童的肥胖盛行率都在上升，且女性较男性更常发生。从1979年到2016年，肥胖成年妇女人数从六千九百万人增加到三亿九千万人，肥胖男性从三千一百万增加到两亿八千一百万。肥胖会带来一系列的并发症，如2型糖尿病、心血管疾病、不孕不育、哮喘、睡眠呼吸暂停综合征等，还与乳腺癌、直肠癌、前列腺癌等恶性肿瘤的发生有关，严重影响了国人的健康和生活质量。Obesity is a very common preventable cause of death and one of the most important public health problems in the 21st century. At present, the prevalence of obesity in both adults and children is on the rise, and it occurs more frequently in women than in men. From 1979 to 2016, the number of obese adult women increased from 69 million to 390 million, and obese men increased from 31 million to 281 million. Obesity will bring a series of complications, such as type 2 diabetes, cardiovascular disease, infertility, asthma, sleep apnea syndrome, etc. It is also related to the occurrence of breast cancer, rectal cancer, prostate cancer and other malignant tumors. It has seriously affected the health and quality of life of Chinese people.
GPHB5是糖蛋白激素家族组成员CGH的亚组，主要由垂体前叶的促肾上腺皮质激素分泌细胞分泌。糖蛋白激素家族成员包括促卵泡激素(FSH)、促黄体素(LH)、促甲状腺激素(TSH)、绒毛膜促性腺激素(CG)，以及2002年被证实为新的糖蛋白激素家族成员CGH。在哺乳动物中，糖蛋白激素对性腺和甲状腺功能起关键作用，促性腺激素能调节生长及卵巢、睾丸的分化，而TSH是机体能量平衡所必须的。GPHB5 is a subgroup of CGH, a member of the glycoprotein hormone family group, and is mainly secreted by ACTH-secreting cells in the anterior pituitary gland. Glycoprotein hormone family members include follicle stimulating hormone (FSH), luteinizing hormone (LH), thyroid stimulating hormone (TSH), chorionic gonadotropin (CG), and CGH, a new glycoprotein hormone family member, was confirmed in 2002 . In mammals, glycoprotein hormones play a key role in gonad and thyroid function. Gonadotropins can regulate growth and differentiation of ovaries and testes, and TSH is necessary for the body's energy balance.
研究表明GPHB5能够和TSH受体(TSHR)结合，并且GPHB5和GPHA2结合形成异质二聚体CGH后，激活TSHR产生cAMP的能力比TSH强4倍。另外，激活脂肪组织TSHR能促进脂肪甘油三酯的水解，进而降低脂肪含量，使体重下降。因此，GPHB5极可能与脂肪组织TSHR结合，刺激甘油三酯水解，可作为一种潜在的减肥减脂分子靶标。Studies have shown that GPHB5 can bind to TSH receptor (TSHR), and after GPHB5 and GPHA2 combine to form a heterodimer CGH, the ability to activate TSHR to produce cAMP is 4 times stronger than TSH. In addition, activating adipose tissue TSHR can promote the hydrolysis of fat triglycerides, thereby reducing fat content and reducing weight. Therefore, GPHB5 is very likely to be combined with TSHR of adipose tissue to stimulate triglyceride hydrolysis, which can be used as a potential molecular target for weight loss and fat reduction.
发明内容Summary of the invention
本发明主要解决的问题在于提供一种减肥减脂，治疗肥胖人群中葡萄糖不耐受或胰岛素抵抗的新机理；在现有研究的基础上，本发明提出了GPHB5糖蛋白激素在肥胖降脂，和治疗肥胖人群中葡萄糖不耐受或胰岛素抵抗中的应用；依据于GPHB5糖蛋白激素在减肥减脂中的作用，揭示一种垂体调控体重和脂肪代谢的新机制。The main problem that the present invention solves is to provide a new mechanism for weight loss and fat reduction, and treatment of glucose intolerance or insulin resistance in obese people; on the basis of existing research, the present invention proposes that the GPHB5 glycoprotein hormone can reduce fat in obesity. And the application in the treatment of glucose intolerance or insulin resistance in obese people; based on the role of GPHB5 glycoprotein hormone in weight loss and fat loss, it reveals a new mechanism for the pituitary to regulate body weight and fat metabolism.
本发明涉及一种具有减肥减脂功效的糖蛋白激素glycoprotein hormone beta  5(GPHB5)。The present invention relates to a glycoprotein hormone beta 5 (GPHB5) with the effect of reducing weight and fat.
一方面，本发明提供了GPHB5糖蛋白激素在制备减少肥胖以及降脂药物中的应用。In one aspect, the present invention provides the application of GPHB5 glycoprotein hormone in the preparation of drugs for reducing obesity and lipid-lowering.
进一步地，所述的GPHB5糖蛋白导致体重下降。Further, the GPHB5 glycoprotein causes weight loss.
进一步地，所述的GPHB5糖蛋白使全身脂肪含量减少，而非脂肪含量无显著变化。Further, the GPHB5 glycoprotein reduces the body fat content, but the non-fat content does not change significantly.
另一方面，本发明提供了GPHB5糖蛋白激素在制备治疗肥胖人群葡萄糖不耐受、胰岛素抵抗或糖尿病的药物中的应用。On the other hand, the present invention provides the application of GPHB5 glycoprotein hormone in the preparation of drugs for the treatment of glucose intolerance, insulin resistance or diabetes in obese people.
所述的GPHB5糖蛋白激素可以为组织中纯化或利用重组表达的GPHB5糖蛋白激素。The GPHB5 glycoprotein hormone can be GPHB5 glycoprotein hormone purified from tissues or recombinantly expressed.
所述的GPHB5糖蛋白激素可以被制备为口服或注射剂型。The GPHB5 glycoprotein hormone can be prepared as an oral or injection dosage form.
另一方面，本发明提供了一种肥胖降脂药物，其特征在于，其中包含GPHB5糖蛋白激素，或表达GPHB5糖蛋白激素的载体或细胞。In another aspect, the present invention provides an obesity-lowering drug, which is characterized in that it contains GPHB5 glycoprotein hormone, or a vector or cell expressing GPHB5 glycoprotein hormone.
进一步地，所述药物效果包括体重下降，或者全身脂肪含量减少而非脂肪含量无显著变化。Further, the drug effect includes weight loss, or reduction in body fat content but no significant change in fat content.
另一方面，本发明提供了一种治疗肥胖人群葡萄糖不耐受或胰岛素抵抗的药物，其特征在于，其中包含GPHB5糖蛋白激素，或表达GPHB5糖蛋白激素的载体或细胞。On the other hand, the present invention provides a medicine for treating glucose intolerance or insulin resistance in obese people, which is characterized in that it contains GPHB5 glycoprotein hormone, or a vector or cell expressing GPHB5 glycoprotein hormone.
所述的药物可以为口服或注射剂型。The medicine can be in the form of oral or injection.
本发明中的GPHB5糖蛋白激素包括各种生物来源的GPHB5糖蛋白激素。The GPHB5 glycoprotein hormone in the present invention includes GPHB5 glycoprotein hormone from various biological sources.
本发明中的GPHB5糖蛋白激素可以是使用各种已知分离纯化方法从组织中纯化得到或者重组表达的GPHB5糖蛋白激素。The GPHB5 glycoprotein hormone in the present invention can be GPHB5 glycoprotein hormone purified from tissues using various known separation and purification methods or recombinantly expressed.
本发明中的GPHB5糖蛋白激素中不排斥其他氨基酸序列的存在，只要不妨碍其行使GPHB5糖蛋白激素功能即可。The GPHB5 glycoprotein hormone in the present invention does not exclude the existence of other amino acid sequences, as long as it does not hinder the function of the GPHB5 glycoprotein hormone.
本发明中的药物中可以含有药学领域/生物领域已知或未知的各种赋形剂、载体、表达细胞，药学领域/生物领域技术人员可以依据现有知识设计相应剂型。The medicine of the present invention may contain various excipients, vectors, and expression cells that are known or unknown in the pharmaceutical field/biological field, and those skilled in the pharmaceutical field/biological field can design corresponding dosage forms based on existing knowledge.
另一方面，本发明提供了一种减少肥胖以及降脂的方法，包括向需要的受试者施用GPHB5糖蛋白激素。In another aspect, the present invention provides a method for reducing obesity and lipid reduction, including administering GPHB5 glycoprotein hormone to a subject in need.
另一方面，本发明提供了一种治疗肥胖人群葡萄糖不耐受、胰岛素抵抗或糖尿病的方法，包括向需要的受试者施用GPHB5糖蛋白激素。In another aspect, the present invention provides a method for treating glucose intolerance, insulin resistance or diabetes in obese people, including administering GPHB5 glycoprotein hormone to subjects in need.
所述的GPHB5糖蛋白激素为组织中纯化或利用重组表达的GPHB5糖蛋白激素。The GPHB5 glycoprotein hormone is a GPHB5 glycoprotein hormone purified from tissues or recombinantly expressed.
本领域的应用和药物中不排斥包含其他有降脂、治疗葡萄糖不耐受或胰岛素抵抗的药物，本领域技术人员可以尝试将这些成分与GPHB5糖蛋白激素组合并验证其效果。The applications and drugs in this field do not exclude other drugs that lower lipids, treat glucose intolerance or insulin resistance. Those skilled in the art can try to combine these ingredients with the GPHB5 glycoprotein hormone and verify its effect.
与现有技术相比，本发明具有的有益效果为：Compared with the prior art, the present invention has the following beneficial effects:
本发明依据于GPHB5糖蛋白激素在减肥减脂中的作用，揭示一种垂体调控体重和脂肪代谢的新机制。本发明通过注射或GPHB5重组激素水平低下的病人，将有助于减肥、增强胰岛素敏感性，并改善血脂代谢。The present invention is based on the role of the GPHB5 glycoprotein hormone in weight loss and fat reduction, and reveals a new mechanism for the pituitary to regulate body weight and fat metabolism. The invention can help lose weight, enhance insulin sensitivity, and improve blood lipid metabolism through injection or patients with low GPHB5 recombinant hormone levels.
附图说明Description of the drawings
图1为GPHB5敲除验证，通过qPCR检测野生型小鼠和敲除鼠的睾丸、大脑、心脏中GPHB5 mRNA表达水平，敲除鼠三种组织中GPHB5mRNA都明显低于野生型，基本不表达。n.d.代表not detectable。Figure 1 shows the GPHB5 knockout verification. GPHB5 mRNA expression levels in the testis, brain, and heart of wild-type mice and knockout mice were detected by qPCR. GPHB5 mRNA in the three tissues of knockout mice was significantly lower than that of wild-type mice, and almost no expression. n.d. stands for not detectable.
图2为GPHB5敲除鼠(-/-)及同窝野生型小鼠(WT)体重增长曲线。Figure 2 shows the body weight growth curve of GPHB5 knockout mice (-/-) and littermate wild-type mice (WT).
图3为GPHB5敲除鼠(-/-)及同窝野生型小鼠(WT)体型的对比。Figure 3 is a comparison of the body types of GPHB5 knockout mice (-/-) and littermate wild-type mice (WT).
图4为GPHB5敲除鼠(-/-)及同窝野生型小鼠(WT)全身脂肪含量(Fat Mass)与非脂肪含量(Lean Mass)的对比。Figure 4 shows the comparison of fat content (Fat Mass) and non-fat content (Lean Mass) of GPHB5 knockout mice (-/-) and litter wild-type mice (WT).
图5为GPHB5敲除鼠(-/-)及同窝野生型小鼠(WT)糖耐量(GTT)的对比。Figure 5 is a comparison of glucose tolerance (GTT) between GPHB5 knockout mice (-/-) and litter wild-type mice (WT).
图6为GPHB5敲除鼠(-/-)及同窝野生型小鼠(WT)胰岛素耐量(ITT)的对比。Figure 6 is a comparison of insulin tolerance (ITT) between GPHB5 knockout mice (-/-) and litter wild-type mice (WT).
图7为表达载体pBudCE4.1-GPHB5/GPHA2质粒的示意图。Figure 7 is a schematic diagram of the expression vector pBudCE4.1-GPHB5/GPHA2 plasmid.
图8为表达载体pcDNA3.1-GPHB5-L-GPHA2质粒的示意图。Figure 8 is a schematic diagram of the expression vector pcDNA3.1-GPHB5-L-GPHA2 plasmid.
图9为Western Blot检测重组GPHB5/GPHA2蛋白。Figure 9 shows the Western Blot detection of recombinant GPHB5/GPHA2 protein.
图10为ELISA检测重组蛋白刺激3T3-L1细胞后细胞中cAMP的变化。Figure 10 shows the change of cAMP in cells after 3T3-L1 cells stimulated by recombinant protein by ELISA.
图中，*代表p＜0.05，差异显著；**代表p＜0.01，差异极显著；***代表p＜0.001，差异极其显著；****代表p＜0.0001，差异非常显著。In the figure, * represents p<0.05, the difference is significant; ** represents p<0.01, the difference is extremely significant; *** represents p<0.001, the difference is extremely significant; **** represents p<0.0001, the difference is very significant.
具体实施方式Detailed ways
为让本领域的技术人员更加清晰直观的了解本发明，下面将结合附图，对本发明作进一步的说明。In order to allow those skilled in the art to understand the present invention more clearly and intuitively, the present invention will be further described below in conjunction with the accompanying drawings.
实施例中，GPHB5糖蛋白激素缺失会导致体重上升，具体地，GPHB5缺失会使全身脂肪含量增加，而非脂肪含量无显著变化，GPHB5缺失会导致葡萄糖不耐受及胰岛素抵抗。In the examples, the lack of GPHB5 glycoprotein hormone will cause weight gain. Specifically, the lack of GPHB5 will increase the body fat content, while the non-fat content will not change significantly. The lack of GPHB5 will cause glucose intolerance and insulin resistance.
应用包括如下步骤：利用从组织中纯化或利用重组表达及纯化的GPHB5蛋白，通过注射或口服，使肥胖病人减肥降脂；或者利用从组织中纯化或利用重组表达及纯化的GPHB5蛋白，通过注射或口服，使肥胖病人改善胰岛素敏感，降低血糖。The application includes the following steps: using GPHB5 protein purified from tissues or recombinantly expressed and purified by injection or oral administration to reduce weight and lipids in obese patients; or using GPHB5 protein purified from tissues or recombinantly expressed and purified by injection Or it can be taken orally to improve insulin sensitivity and lower blood sugar in obese patients.
实施例1 GPHB5敲除鼠的构建、验证和相关指标分析Example 1 Construction, verification and related index analysis of GPHB5 knockout mice
小鼠GPHB5基因的Genbank号为NM_175644.3，人GPHB5基因的Genbank号为NM_145171.4，GPHB5基因全长约为3900bp。所用的GPHB5敲除鼠是委托赛业生物科技有限公司，通过CRISPR/Cas9基因敲除技术，敲除GPHB5基因的2号及3号外显子，敲除3340bp碱基得到2只GPHB5敲除鼠，标记为line1和line2。通过DNA测序检测可能脱靶的前10个位点，结果表明line1和line2均没有脱靶，说明GPHB5敲除鼠构建成功。The Genbank number of the mouse GPHB5 gene is NM_175644.3, the Genbank number of the human GPHB5 gene is NM_145171.4, and the full length of the GPHB5 gene is about 3900bp. The GPHB5 knockout mice used were commissioned by Saiye Biotechnology Co., Ltd., through the CRISPR/Cas9 gene knockout technology, the 2 and 3 exons of the GPHB5 gene were knocked out, and the 3340bp base was knocked out to obtain 2 GPHB5 knockout mice. Labeled as line1 and line2. The first 10 sites that may be off-target were detected by DNA sequencing, and the results showed that line1 and line2 were not off-target, indicating that the GPHB5 knockout mouse was successfully constructed.
通过杂合子与杂合子配繁，得到GPHB5敲除鼠和同窝野生型小鼠。取GPHB5敲除鼠及野生型小鼠的睾丸、大脑、心脏，检测GPHB5mRNA水平，如图1所示，结果表明GPHB5敲除鼠基本不表达GPHB5mRNA。Through heterozygous and heterozygous mating, GPHB5 knockout mice and littermate wild-type mice were obtained. The testes, brains, and hearts of GPHB5 knockout mice and wild-type mice were taken to detect GPHB5mRNA levels. As shown in Figure 1, the results showed that GPHB5 knockout mice basically did not express GPHB5mRNA.
GPHB5敲除鼠与同窝野生型小鼠体重比较：Comparison of body weight between GPHB5 knockout mice and wild-type mice in the same litter:
SPF屏障环境中饲养小鼠，同窝雄鼠放同一笼，每笼3-4只，喂相同的饲料和无菌水。从小鼠6周龄成熟后开始记录体重，每周记录一次，如图2和图3所示。从鼠龄18周之后，GPHB5敲除鼠体重明显高于同窝野生型小鼠，31周时GPHB5敲除鼠体重比野生型小鼠多6g，说明GPHB5缺失会导致肥胖。Mice were reared in an SPF barrier environment. Male mice in the same litter were placed in the same cage, with 3-4 mice per cage, and fed with the same feed and sterile water. The body weight of the mice was recorded after 6 weeks of age and was recorded once a week, as shown in Figure 2 and Figure 3. After 18 weeks of age, the body weight of GPHB5 knockout mice was significantly higher than that of wild-type mice in the same litter. At 31 weeks, the body weight of GPHB5 knockout mice was 6g more than that of wild-type mice, indicating that GPHB5 deletion can lead to obesity.
全身脂肪含量与非脂肪含量分析：Analysis of body fat content and non-fat content:
采用汇佳生物股份有限公司的体成分分析仪Echo MRI，检测GPHB5敲除鼠与同窝野生型小鼠的全身脂肪含量与非脂肪含量。如图4所示，GPHB5敲除鼠的全身脂肪含量明显高于同窝野生型小鼠，而非脂肪含量基本一致，表明GPHB5敲除鼠体重的增加主要来源于脂肪的累积。The body composition analyzer Echo MRI of Huijia Biological Co., Ltd. was used to detect the body fat content and non-fat content of GPHB5 knockout mice and littermated wild-type mice. As shown in Figure 4, the body fat content of GPHB5 knockout mice was significantly higher than that of wild-type mice in the same litter, while the non-fat content was basically the same, indicating that the increase in body weight of GPHB5 knockout mice was mainly due to the accumulation of fat.
糖耐量分析：Glucose tolerance analysis:
GPHB5敲除鼠和同窝野生型小鼠禁食12小时，罗氏血糖仪测空腹血糖值。按2g/kg体重，腹腔注射20％葡萄糖，罗氏血糖仪测注射后15min、30min、60min、90min、120min、150min小鼠的血糖值，如图5所示，结果表明GPHB5敲除鼠的糖耐量明显低于同窝野生型小鼠。GPHB5 knockout mice and wild-type mice in the same litter were fasted for 12 hours, and the fasting blood glucose was measured by Roche blood glucose meter. According to 2g/kg body weight, 20% glucose was injected intraperitoneally, and the blood glucose level of mice 15min, 30min, 60min, 90min, 120min, 150min after injection was measured by Roche blood glucose meter, as shown in Figure 5, the results showed the glucose tolerance of GPHB5 knockout mice It is significantly lower than that of wild-type mice in the same litter.
胰岛素耐量分析：Insulin tolerance analysis:
GPHB5敲除鼠和同窝野生型小鼠禁食6小时后，用罗氏血糖仪测空腹血糖，按0.8IU/kg体重，腹腔注射0.2IU/ml的胰岛素稀释液，罗氏血糖仪测注射后15min、30min、45min、60min、75min、90min小鼠的血糖值，并以注射前测得的血糖值为100％作胰岛素耐量曲线，如图6所示。结果表明GPHB5敲除鼠对胰岛素不敏感。After fasting for 6 hours in GPHB5 knockout mice and littermate wild-type mice, use Roche blood glucose meter to measure fasting blood glucose. According to 0.8IU/kg body weight, intraperitoneal injection of 0.2IU/ml insulin diluent, Roche blood glucose meter measures 15min after injection , 30min, 45min, 60min, 75min, 90min mice blood glucose values, and the blood glucose value measured before injection is 100% as the insulin tolerance curve, as shown in Figure 6. The results show that GPHB5 knockout mice are not sensitive to insulin.
实施例2 重组GPHB5/GPHA2蛋白在哺乳动物细胞中的表达和功能分析Example 2 Expression and functional analysis of recombinant GPHB5/GPHA2 protein in mammalian cells
表达载体pBudCE4.1-GPHB5/GPHA2质粒的构建：Construction of expression vector pBudCE4.1-GPHB5/GPHA2 plasmid:
GPHB5和GPHA2全基因序列由南京金斯瑞公司合成，并进行密码子人源化优化。目的基因克隆至pBudCE4.1载体中，其中GPHB5基因克隆至载体的Hind III/BamH I位点之间，GPHA2基因克隆至载体的Not I/Xho I位点之间，如图7所示。将pBudCE4.1-GPHB5/GPHA2质粒转化至MJ109大肠杆菌中，挑取单克隆进行质粒酶切鉴定以及DNA测序方法验证插入序列的正确性。pBudCE4.1质粒为哺乳动物细胞表达双基因常用的载体，载体上携带有His标签、V5标签和myc标签序列，根据基因插入位点显示，表达重组蛋白后GPHB5蛋白将携带myc和His标签，GPHA2蛋白将携带V5和His标签，可用于纯化和检测。The complete gene sequences of GPHB5 and GPHA2 were synthesized by Nanjing GenScript Company and optimized by humanization of codons. The target gene was cloned into the pBudCE4.1 vector, where the GPHB5 gene was cloned between the Hind III/BamH I sites of the vector, and the GPHA2 gene was cloned between the Not I/Xho I sites of the vector, as shown in Figure 7. The pBudCE4.1-GPHB5/GPHA2 plasmid was transformed into MJ109 Escherichia coli, and a single clone was picked for plasmid identification and DNA sequencing to verify the correctness of the inserted sequence. pBudCE4.1 plasmid is a commonly used vector for expressing double genes in mammalian cells. The vector carries His tag, V5 tag and myc tag sequence. According to the gene insertion site, GPHB5 protein will carry myc and His tag after expressing recombinant protein, GPHA2 The protein will carry V5 and His tags, which can be used for purification and detection.
表达载体pcDNA3.1-GPHB5-L-GPHA2质粒的构建：Construction of expression vector pcDNA3.1-GPHB5-L-GPHA2 plasmid:
合成含有GPHB5的信号肽以及成熟肽段、连接肽L、GPHA2的成熟肽和His标签的融合基因，克隆至pCDNA3.1载体的EcoR I/Not I位点之间，如图8所示。其中，连接肽L的蛋白序列有两种：常规连接肽GGGGSGGGGSGGGGS，以及能够延长重组蛋白半衰期的CTP肽段PRFQDSSSSKAPPPSLPSPSRLPGPSDTPILPQ。The fusion gene containing the signal peptide of GPHB5 and the mature peptide, the connecting peptide L, the mature peptide of GPHA2 and the His tag was synthesized, and cloned into the EcoR I/Not I site of the pCDNA3.1 vector, as shown in Figure 8. Among them, there are two protein sequences of connecting peptide L: the conventional connecting peptide GGGGSGGGGSGGGGS, and the CTP peptide PRFQDSSSSKAPPPSLPSPSRLPGPSDTPILPQ, which can extend the half-life of the recombinant protein.
重组GPHB5/GPHA2蛋白在哺乳动物细胞中的表达及功能分析：Expression and function analysis of recombinant GPHB5/GPHA2 protein in mammalian cells:
将构建的表达载体pBudCE4.1-GPHB5/GPHA2和表达载体pcDNA3.1-GPHB5-L-GPHA2转染至哺乳动物细胞中，包括HEK 293T细胞、293F细胞、CHO细胞。图9为表达载体pBudCE4.1-GPHB5/GPHA2转染293T细胞后继续培养2天后，取培养基上清进行Western Blot实验的检测结果，表明GPHB5/GPHA2重组蛋白表达成功。对含有重组蛋白的培养基进行超滤管浓缩5倍，His标签纯化试剂盒进行纯化后，用于刺激3T3-L1细胞20min、60min、90min，ELISA检测细胞内cAMP水平，发现与对照组相比，表达出来的GPHB5/GPHA2重组蛋白刺激20min和60min后cAMP上升，如图10所示，说明表达出来的重组蛋白具有活性。The constructed expression vector pBudCE4.1-GPHB5/GPHA2 and the expression vector pcDNA3.1-GPHB5-L-GPHA2 were transfected into mammalian cells, including HEK 293T cells, 293F cells, and CHO cells. Figure 9 shows the result of Western Blot experiment with the culture supernatant after transfection of the expression vector pBudCE4.1-GPHB5/GPHA2 into 293T cells, which indicates that the GPHB5/GPHA2 recombinant protein was successfully expressed. The culture medium containing the recombinant protein was concentrated by ultrafiltration tube 5 times. After purification by the His tag purification kit, it was used to stimulate 3T3-L1 cells for 20 min, 60 min, and 90 min. The intracellular cAMP level was detected by ELISA, and it was found to be compared with the control group. , The expressed GPHB5/GPHA2 recombinant protein stimulates cAMP to increase after 20min and 60min, as shown in Figure 10, indicating that the expressed recombinant protein is active.
上述对实施例的描述是为便于该技术领域的普通技术人员能理解和应用本发明。熟悉本领域技术的人员显然可以容易地对这些实施例做出各种修改，并把在此说明的一般原理应用到其他实施例中而不必经过创造性的劳动。因此，本发明不限于这里的实施例，本领域技术人员根据本发明的揭示，对于本发明做出的改进和修改都应该在本发明的保护范围之内。The above description of the embodiments is to facilitate those of ordinary skill in the technical field to understand and apply the present invention. It is obvious that those skilled in the art can easily make various modifications to these embodiments, and apply the general principles described here to other embodiments without creative work. Therefore, the present invention is not limited to the embodiments herein. Based on the disclosure of the present invention, those skilled in the art make improvements and modifications to the present invention should all fall within the protection scope of the present invention.

Claims (13)

  1. GPHB5糖蛋白激素在制备减少肥胖以及降脂药物中的应用。Application of GPHB5 glycoprotein hormone in the preparation of drugs for reducing obesity and lipid-lowering.
  2. 根据权利要求1所述的应用，其中所述的GPHB5糖蛋白激素导致体重下降。The use according to claim 1, wherein the GPHB5 glycoprotein hormone causes weight loss.
  3. 根据权利要求1所述的应用，其中所述的GPHB5糖蛋白激素使全身脂肪含量减少，而非脂肪含量无显著变化。The use according to claim 1, wherein the GPHB5 glycoprotein hormone reduces the body fat content, but the non-fat content does not change significantly.
  4. GPHB5糖蛋白激素在制备治疗肥胖人群葡萄糖不耐受、胰岛素抵抗或糖尿病的药物中的应用。The application of GPHB5 glycoprotein hormone in the preparation of drugs for the treatment of glucose intolerance, insulin resistance or diabetes in obese people.
  5. 根据权利要求1-4任一项所述的应用，其中所述的GPHB5糖蛋白激素为组织中纯化或利用重组表达的GPHB5糖蛋白激素。The application according to any one of claims 1 to 4, wherein the GPHB5 glycoprotein hormone is GPHB5 glycoprotein hormone purified from tissues or recombinantly expressed.
  6. 根据权利要求1-5任一项所述的应用，其中所述的GPHB5糖蛋白激素被制备为口服或注射剂型。The use according to any one of claims 1-5, wherein the GPHB5 glycoprotein hormone is prepared as an oral or injection dosage form.
  7. 一种肥胖降脂药物，其特征在于，其中包含GPHB5糖蛋白激素，或表达GPHB5糖蛋白激素的载体或细胞。An obesity-lowering drug is characterized in that it contains GPHB5 glycoprotein hormone, or a carrier or cell expressing GPHB5 glycoprotein hormone.
  8. 根据权利要求7所述的药物，其中药物效果包括体重下降，或者全身脂肪含量减少而非脂肪含量无显著变化。The medicine according to claim 7, wherein the effect of the medicine includes weight loss, or reduction of body fat content but no significant change in fat content.
  9. 一种治疗肥胖人群葡萄糖不耐受或胰岛素抵抗的药物，其特征在于，其中包含GPHB5糖蛋白激素，或表达GPHB5糖蛋白激素的载体或细胞。A medicine for treating glucose intolerance or insulin resistance in obese people, which is characterized in that it contains GPHB5 glycoprotein hormone, or a carrier or cell expressing GPHB5 glycoprotein hormone.
  10. 根据权利要求7-9任一项所述的药物，其为口服或注射剂型。The medicine according to any one of claims 7-9, which is an oral or injection dosage form.
  11. 一种减少肥胖以及降脂的方法，其特征在于，包括向需要的受试者施用GPHB5糖蛋白激素。A method for reducing obesity and lipid-lowering is characterized by comprising administering GPHB5 glycoprotein hormone to a subject in need.
  12. 一种治疗肥胖人群葡萄糖不耐受、胰岛素抵抗或糖尿病的方法，其特征在于，包括向需要的受试者施用GPHB5糖蛋白激素。A method for treating glucose intolerance, insulin resistance or diabetes in obese people is characterized by comprising administering GPHB5 glycoprotein hormone to subjects in need.
  13. 根据权利要求11或12所述的方法，其中所述的GPHB5糖蛋白激素为组织中纯化或利用重组表达的GPHB5糖蛋白激素。The method according to claim 11 or 12, wherein the GPHB5 glycoprotein hormone is purified or recombinantly expressed GPHB5 glycoprotein hormone in tissues.
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Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20020160953A1 (en) * 2000-04-25 2002-10-31 Holloway James L. Mammalian glycoprotein hormone-1
WO2007005320A2 (en) * 2005-06-29 2007-01-11 Zymogenetics, Inc. Methods of delivering corticotroph-derived glycoprotein hormone

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20050096264A1 (en) * 2003-02-25 2005-05-05 Macdonald Lynn Methods for treating obesity and related conditions with glycoprotein hormone beta family hormones
CN110904046B (en) * 2018-09-18 2021-10-08 中国农业大学 Application of ISLR gene in preparation of medicine for treating obesity and improving insulin resistance

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20020160953A1 (en) * 2000-04-25 2002-10-31 Holloway James L. Mammalian glycoprotein hormone-1
WO2007005320A2 (en) * 2005-06-29 2007-01-11 Zymogenetics, Inc. Methods of delivering corticotroph-derived glycoprotein hormone

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
SHANNON L. OKADA*, JEFF L. ELLSWORTH, DIANE M. DURNAM, HARALD S. HAUGEN, JAMES L. HOLLOWAY, MERIDETH L. KELLEY, KATHERINE E. LEWIS: "Corticotroph Localization and Thyroid-Stimulating Hormone Receptor Binding Properties of Corticotroph-derived Glycoprotein Hormone", MOLECULAR ENDOCRINOLOGY, THE ENDOCRINE SOCIETY, US, vol. 20, no. 2, 1 January 2006 (2006-01-01), US , pages 414 - 425, XP009135306, ISSN: 0888-8809 *

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