Composition comprising GLP-1 receptor agonist and glucagon receptor agonist and uses thereof
一种胰高血糖素、受体激动剂的技术,应用在减肥和/或降脂领域,能够解决活性差、半衰期短等问题,达到促进体重减轻、食物摄取减少、减少体重增加的效果
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Embodiment 1
[0090] Example 1: Preparation of PB-119 (a conjugate of PB-105 and mPEG-ppMAL-23KD)
[0091] Weigh 10mg of PB-105 (purchased from Chengdu Capgemini Biomedical Technology Development Co., Ltd.) and 65mg of mPEG23KD-ppMAL (produced by Paige Biopharmaceutical Company) in a 50mL glass container bottle (the molar ratio of PB-105 to PEG is 1:1.2 ), dissolved in 5ml of 0.2M PBS buffer solution with pH 6.5, and stirred at room temperature (25°C) for 1 hour, then adjusted the pH to 4.0 with hydrochloric acid to terminate the reaction, and placed at 4°C for purification.
[0092] Dilute the above reaction solution 5 times with ultrapure water, adjust the pH to 4.0 with acetic acid, and load the sample on the cation exchange of 10 mL Macrocap SP equilibrated with buffer A (20 mM pH 4.0 HAc-NaAc) for 3-4 column volumes. Column, after loading, use buffer A (20mM pH 4.0 HAc-NaAc) to elute excess PEG, then use eluent B (20mM pH4.0 HAc-NaAc+1M NaCl) to elute 10 column volume gradients When t...
Embodiment 2
[0093] Example 2: Preparation of PB-120 (a conjugate of PB-105 and mPEG-ppMAL-25KD)
[0094] Weigh 10mg of PB-105 (purchased from Chengdu Capgemini Biomedical Technology Development Co., Ltd.) and 72mg of mPEG25KD-ppMAL (produced by Paige Biopharmaceutical Company) in a 50mL glass container bottle (the molar ratio of PB-105 to PEG is 1:1.2 ), dissolved in 5ml of 0.2M PBS buffer solution with pH 6.5, and stirred at room temperature (25°C) for 1 hour, then adjusted the pH to 4.0 with hydrochloric acid to terminate the reaction, and placed at 4°C for purification.
[0095] Dilute the above reaction solution 5 times with ultrapure water, adjust the pH to 4.0 with acetic acid, and load the sample on the cation exchange of 10 mL Macrocap SP equilibrated with buffer A (20 mM pH 4.0 HAc-NaAc) for 3-4 column volumes. Column, after loading, use buffer A (20mM pH 4.0 HAc-NaAc) to elute excess PEG, then use eluent B (20mM pH4.0 HAc-NaAc+1M NaCl) to elute 10 column volume gradients When t...
Embodiment 3
[0096] Example 3: Preparation of PB-107 (a conjugate of PB-105 and mPEG-ppMAL-30KD)
[0097] Weigh 10mg of PB-105 (purchased from Chengdu Capgemini Biomedical Technology Development Co., Ltd.) and 86mg of mPEG30KD-ppMAL (produced by Paige Biopharmaceutical Company) in a 50mL glass container bottle (the molar ratio of PB-105 to PEG is 1:1.2 ), dissolved in 5ml of 0.2M PBS buffer at pH 6.5, and stirred at room temperature (25°C) for 1 hour, then adjusted to pH 4.0 with hydrochloric acid to terminate the reaction, and placed at 4°C for purification.
[0098] Dilute the above reaction solution 5 times with ultrapure water, adjust the pH to 4.0 with acetic acid, and load the sample on the cation exchange of 10 mL Macrocap SP equilibrated with buffer A (20 mM pH 4.0 HAc-NaAc) for 3-4 column volumes. Column, after loading, use buffer A (20mM pH 4.0 HAc-NaAc) to elute excess PEG, then use eluent B (20mM pH4.0 HAc-NaAc+1M NaCl) to elute 10 column volume gradients When the elution reac...
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