Smeglutide microsphere preparation and preparation method thereof

Abstract

The invention discloses a semeglutide microsphere preparation and a preparation method thereof, and belongs to the technical field of medicine synthesis. The preparation method comprises the following steps: adding polyvinyl alcohol into water, heating to 70-80 DEG C for dissolving, and cooling to obtain a PVA solution; the preparation method comprises the following steps: adding a PEG copolymer into an organic solvent, dissolving, filtering, adding the semeglutide into filtrate, and dissolving to obtain a semeglutide solution; taking a semeglutide solution as a dispersion phase, taking a PVA (Polyvinyl Alcohol) solution as a mobile phase, and obtaining a semeglutide emulsion through a membrane emulsification method; the preparation method comprises the following steps: carrying out homogenization emulsification treatment on a semeglutide emulsion, stirring the treated emulsion, and carrying out microsphere curing, washing and drying to obtain the semeglutide microsphere preparation. The preparation prepared by using the biodegradable high polymer material as the auxiliary material can effectively prolong the in-vivo action time of the semeglutide, reduce the number of times of medicine injection, improve the adaptability of patients, facilitate clinical use and greatly improve the use experience of the patients.

Description

Technical Field

[0001] The invention belongs to the technical field of drug synthesis, and particularly relates to a semaglutide microsphere preparation and a preparation method thereof. Background Art

[0002] Obesity seriously troubles people's lives and health. The World Health Organization has pointed out that overweight and obesity are the fifth largest risk of death in the world. At present, the number of overweight people in my country has reached 200 million, and the number of obese people has exceeded 90 million, of which 12 million are severely obese.

[0003] Semaglutide was originally developed by Novo Nordisk as a hypoglycemic drug for the treatment of type 2 diabetes. In June 2021, the FDA approved Semaglutide for marketing as a weight loss drug (trade name Wegovy). CN 1011330882 reported that Semaglutide helps the pancreas release an appropriate amount of insulin to control blood sugar when blood sugar is high. It acts on the brain area that regulates appetite and food intake, and reduces appetite and weight by slowing down gastric emptying.

[0004] Semaglutide needs to be used for a long time to exert its therapeutic benefits, and this type of drug requires patients to inject it every day or every week, and the injection needs to be stored at 2-8°C, which brings great inconvenience to patients. Even the oral version currently under development also requires frequent administration. Therefore, maintaining continuous plasma drug levels by improving drug delivery technology is important for better treatment of diabetes, weight loss, and prevention of cardiovascular disease. Summary of the invention

[0005] In view of the problem that frequent administration of semaglutide in the prior art brings inconvenience to patients, the present invention provides a semaglutide microsphere preparation and a preparation method thereof. The preparation prepared using biodegradable polymer materials as excipients can effectively prolong the action time of semaglutide in the body, reduce the number of drug injections, improve the patient's adaptability, and is more convenient for clinical use, greatly improving the patient's usage experience.

[0006] The present invention is achieved through the following technical solutions: A method for preparing a semaglutide microsphere preparation comprises the following steps: (1) Add polyvinyl alcohol to water, heat to 70-80°C to dissolve, and cool to obtain a PVA solution; (2) adding the PEG copolymer to an organic solvent to dissolve, filtering, and adding semaglutide to the filtrate to dissolve to obtain a semaglutide solution; (3) Using semaglutide solution as dispersed phase and PVA solution as mobile phase, semaglutide emulsion was obtained by membrane emulsification method; (4) The semaglutide emulsion is homogenized and emulsified, the emulsion is stirred, the microspheres are solidified, washed, and dried to obtain a semaglutide microsphere preparation.

[0007] Furthermore, in step (1), the concentration of the PVA solution is 0.2-5%; and the weight average molecular weight of the polyvinyl alcohol is 84,000-124,000.

[0008] Furthermore, the PEG copolymer in step (2) is one of PLGA-PEG-PLGA triblock copolymer, PEG-PLLA diblock copolymer, mPEG-PLGA diblock copolymer, and mPEG-PLLA diblock copolymer; and the organic solvent is one of ethyl acetate, dichloromethane, chloroform, and tetrahydrofuran.

[0009] Furthermore, the molar ratio of lactide to glycolide in the PLGA-PEG-PLGA triblock copolymer and the mPEG-PLGA diblock copolymer is 50-90:50-10; and the molecular weight of PEG and mPEG in the PEG copolymer is 400-2000.

[0010] Furthermore, the molar ratio of lactide to glycolide in the PLGA-PEG-PLGA triblock copolymer and the mPEG-PLGA diblock copolymer is 75-85:25-15; the molecular weight of mPEG in the mPEG-PLGA diblock copolymer and the mPEG-PLLA diblock copolymer is 800-1000.

[0011] Furthermore, in step (2), the mass ratio of the PEG copolymer to semaglutide is 1:0.5-2; and the amount of the organic solvent used is 8-10 times the mass of the PEG copolymer.

[0012] Furthermore, in step (3), the semaglutide solution is poured into a membrane emulsification tank, nitrogen is passed through and pressurized to 0.05-0.25 MPa, and the dispersed phase is extruded through the membrane tube. The dispersed phase forms spherical droplets on the membrane surface, which slowly detach from the membrane surface under the action of the fast-flowing PVA solution until no droplets are formed on the membrane surface, and the nitrogen is turned off to generate a semaglutide emulsion. In step (4), the drying method is vacuum freeze-drying.

[0013] Furthermore, the membrane tube in step (3) is made of SPG membrane or stainless steel, and the pore size of the membrane tube is 1-200 μm.

[0014] Furthermore, the homogenization emulsification conditions in step (4) are 15000-20000 rpm for 20-40 min.

[0015] In the present invention, the semaglutide microsphere preparation is prepared by the preparation method.

[0016] Beneficial effects achieved by the present invention: The present invention discloses a method for preparing a semaglutide microsphere preparation. The pegylation modification technology couples polyethylene glycol with a polyester polymer material through a covalent bond, thereby improving the physical and chemical properties of the auxiliary material, making the preparation process of the microsphere easier to repeat and control, and the sustained-release effect of the preparation is better.

[0017] The sustainable treatment period of semaglutide preparations prepared with PEGylated copolymers as excipients can be extended to 30 to 80 days. The long-acting sustained-release direct use method is more convenient, enriching the use period of semaglutide preparations and laying a more solid foundation for the development of new indications for semaglutide. The semaglutide microsphere preparation prepared by the present invention can effectively prolong the action time of semaglutide in the body, reduce the number of drug injections, improve the adaptability of patients, is more convenient for clinical use, and greatly improves the patient's use experience. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 This is a scanning electron micrograph of the semaglutide microspheres prepared in Example 1; Figure 2 This is the particle size distribution diagram of semaglutide microspheres prepared in Example 1; Figure 3 This is a graph showing the release rate change of the semaglutide microspheres prepared in Example 1 in an in vitro degradation experiment; Figure 4 This is a graph showing the change in release rate per unit time of the in vitro degradation experiment of semaglutide microspheres prepared in Example 1. DETAILED DESCRIPTION

[0019] The above contents of the present invention are further described in detail below through examples, but this should not be understood as the scope of the above subject matter of the present invention being limited to the following examples, and all technologies implemented based on the above contents of the present invention belong to the scope of the present invention.

[0020] Example 1 (1) Add polyvinyl alcohol (weight average molecular weight: 108,000) to water under stirring, heat to 75°C to dissolve, and cool to room temperature to obtain a PVA solution with a mass concentration of 1%; (2) Add PLGA-PEG-PLGA triblock copolymer (PLGA monomer ratio of 85:15, PEG segment of 800, total molecular weight of 50379) to tetrahydrofuran (8 times the mass of PLGA-PEG-PLGA triblock copolymer) to dissolve, filter to remove impurities and sterilize, add semaglutide to the filtrate, stir to dissolve, and obtain semaglutide solution. The mass ratio of PLGA-PEG-PLGA triblock copolymer to semaglutide is 1:1; (3) The semaglutide solution was used as the dispersed phase and the PVA solution as the mobile phase. The semaglutide solution was poured into the membrane emulsification tank and pressurized to 0.20-0.25 MPa with nitrogen. The dispersed phase was extruded through a stainless steel membrane tube (a two-stage tube with pore sizes of 50 μm and 30 μm). The dispersed phase formed spherical droplets on the membrane surface and slowly separated from the membrane surface under the flow of the PVA solution. The pressurization pressure was controlled at 0.20-0.25 MPa until no droplets were formed on the membrane surface. The nitrogen was then turned off to generate a semaglutide emulsion. (4) The semaglutide emulsion was homogenized and emulsified (18000 rpm, 30 min). The treated emulsion was stirred at 50±3°C and 50 rpm. The microspheres were solidified, washed with water, and freeze-dried under vacuum conditions to obtain a semaglutide microsphere preparation. The drug loading of the semaglutide microsphere preparation was found to be 32%, and the encapsulation efficiency was 91%.

[0021] The scanning electron microscopy image of the semaglutide microsphere preparation prepared in Example 1 is as follows Figure 1 The particle size distribution diagram is shown in Figure 2 As shown, the particle size of semaglutide microsphere preparation is D10: 13 μm, D90: 85 μm, and span value: 0.60.

[0022] Example 2 (1) Add polyvinyl alcohol (weight average molecular weight: 108,000) to water under stirring, heat to 75°C to dissolve, and cool to room temperature to obtain a PVA solution with a mass concentration of 1%; (2) Add mPEG-PLGA diblock copolymer (PLGA monomer ratio of 85:15, PEG segment of 1000, total molecular weight of 51082) to tetrahydrofuran (9 times the mass of mPEG-PLGA diblock copolymer) to dissolve, filter to remove impurities and sterilize, add semaglutide to the filtrate, stir to dissolve, and obtain semaglutide solution. The mass ratio of mPEG-PLGA diblock copolymer to semaglutide is 1:1; (3) The semaglutide solution was used as the dispersed phase and the PVA solution as the mobile phase. The semaglutide solution was poured into the membrane emulsification tank and pressurized to 0.20-0.25 MPa with nitrogen. The dispersed phase was extruded through a stainless steel membrane tube (a two-stage tube with pore sizes of 50 μm and 30 μm). The dispersed phase formed spherical droplets on the membrane surface and slowly separated from the membrane surface under the flow of the PVA solution. The pressurization pressure was controlled at 0.20-0.25 MPa until no droplets were formed on the membrane surface. The nitrogen was then turned off to generate a semaglutide emulsion. (4) The semaglutide emulsion was homogenized and emulsified (18000 rpm, 30 min), and the treated emulsion was stirred at 50±3°C and 50 rpm, the microspheres were solidified, washed with water, and freeze-dried under vacuum conditions to obtain a semaglutide microsphere preparation; after testing, the drug loading of the semaglutide microsphere preparation was 31%, the encapsulation efficiency was 90%, the particle size of the sample was D10: 12 μm, D90: 82 μm, and the span value was 0.63.

[0023] Example 3 (1) Add polyvinyl alcohol (weight average molecular weight: 108,000) to water under stirring, heat to 75°C to dissolve, and cool to room temperature to obtain a PVA solution with a mass concentration of 1%; (2) Add mPEG-PLA diblock copolymer (mPEG segment is 1000, total molecular weight is 47853) to tetrahydrofuran (10 times the mass of mPEG-PLA diblock copolymer) to dissolve, filter to remove impurities and sterilize, add semaglutide to the filtrate, stir to dissolve, and obtain semaglutide solution, the mass ratio of mPEG-PLA diblock copolymer to semaglutide is 1:1; (3) The semaglutide solution was used as the dispersed phase and the PVA solution as the mobile phase. The semaglutide solution was poured into the membrane emulsification tank and pressurized to 0.20-0.25 MPa with nitrogen. The dispersed phase was extruded through a stainless steel membrane tube (a two-stage tube with pore sizes of 50 μm and 30 μm). The dispersed phase formed spherical droplets on the membrane surface and slowly separated from the membrane surface under the flow of the PVA solution. The pressurization pressure was controlled at 0.20-0.25 MPa until no droplets were formed on the membrane surface. The nitrogen was then turned off to generate a semaglutide emulsion. (4) The semaglutide emulsion was homogenized and emulsified (18000 rpm, 30 min), and the treated emulsion was stirred at 50±3°C and 50 rpm. The microspheres were solidified, washed with water, and freeze-dried under vacuum conditions to obtain semaglutide microsphere preparations. After testing, the drug loading of the semaglutide microsphere preparation was 31%, the encapsulation efficiency was 88%, the particle size of the sample was D10: 15 μm, D90: 96 μm, and the span value was 0.67.

[0024] Comparative Example 1 (1) Add polyvinyl alcohol (weight average molecular weight: 108,000) to water under stirring, heat to 75°C to dissolve, and cool to room temperature to obtain a PVA solution with a mass concentration of 1%; (2) adding PLGA copolymer (PLGA monomer ratio of 85:15, total molecular weight of 48935) to tetrahydrofuran (8 times the mass of PLGA copolymer) to dissolve, filtering to remove impurities and sterilizing, adding semaglutide to the filtrate, stirring to dissolve, and obtaining semaglutide solution, wherein the mass ratio of PLGA copolymer to semaglutide is 1:1; (3) The semaglutide solution was used as the dispersed phase and the PVA solution as the mobile phase. The semaglutide solution was poured into the membrane emulsification tank and pressurized to 0.20-0.25 MPa with nitrogen. The dispersed phase was extruded through a stainless steel membrane tube (a two-stage tube with pore sizes of 50 μm and 30 μm). The dispersed phase formed spherical droplets on the membrane surface and slowly separated from the membrane surface under the flow of the PVA solution. The pressurization pressure was controlled at 0.20-0.25 MPa until no droplets were formed on the membrane surface. The nitrogen was then turned off to generate a semaglutide emulsion. (4) The semaglutide emulsion was homogenized and emulsified (18000 rpm, 30 min), and the treated emulsion was stirred at 50±3°C and 50 rpm. The microspheres were solidified, washed with water, and freeze-dried under vacuum conditions to obtain semaglutide microsphere preparations. After testing, the drug loading of the semaglutide microsphere preparation was 26%, the encapsulation efficiency was 65%, the particle size of the sample was D10: 16 μm, D90: 129 μm, and the span value was 0.98.

[0025] From the comparison of the above embodiments and comparative examples, it can be seen that the semaglutide microspheres prepared using the modified materials PLGA-PEG-PLGA triblock copolymer, mPEG-PLGA diblock copolymer, and mPEG-PLA diblock copolymer (relative to PLGA copolymer) have certain advantages in drug loading, encapsulation efficiency and microsphere particle size, and the microspheres have better sphericity under a microscope. This is because the PEG-modified biodegradable materials (such as PLGA-PEG-PLGA triblock copolymers, mPEG-PLGA diblock copolymers, and mPEG-PLA diblock copolymers) have improved hydrophilic and hydrophobic properties after adding PEG segments, making the material more stable during the membrane emulsification spheronization process. In addition, tetrahydrofuran, a solvent miscible with water, is used in this process, so that after the microspheres form a layer, the solvent tetrahydrofuran that can dissolve the material is diluted, and the initial morphology of the microspheres is retained to a greater extent during the homogenization process. At the same time, the drug loading and encapsulation rate of the microspheres are guaranteed, and the advantages of the membrane emulsification process are maximized, so that the particle size distribution of the obtained microspheres is controllable and the batch difference is small.

[0026] In vitro degradation experiment of semaglutide microsphere preparation: 20 mg of semaglutide microspheres prepared in Example 1 were accurately weighed and placed in a 50 ml sealed test tube, 20 ml of release medium (phosphate buffer solution with a pH of 7.4±0.2) was added, the lid was screwed and sealed and mixed, and then the test tube was placed on a 37±0.5°C constant temperature shaker and shaken at a speed of 100 rpm. Samples were taken at 1 day, 2 days, 4 days, 8 days, 12 days, 24 days, 36 days, 48 ​​days and 72 days after placement. Before sampling, the test tube was slowly rotated to evenly suspend the solution, and the solution was allowed to stand for at least 10 minutes to allow the microspheres to fully settle. Samples were taken at about 2 cm below the liquid surface, 1 ml of each solution was accurately measured, and an equal volume of isothermal release medium was added to ensure that the release was in a trough state. The collected samples were filtered through a 0.45 μm microporous filter membrane, and the filtrate was taken as the test solution. 20 μl of the sample was taken from the HPLC to determine the released semaglutide content and calculate the release rate. Figure 3 As shown, the release rate per unit time of the in vitro degradation experiment of semaglutide microspheres prepared in Example 1 is shown in Figure 4 As shown, the release rate per unit time of the in vitro degradation experiment of the semaglutide microspheres prepared in Example 1 is shown in Table 1.

[0027] Table 1 Table of release rate per unit time in the in vitro degradation experiment of semaglutide microspheres prepared in Example 1 Depend on Figure 3 , Figure 4 As shown in Table 1, the semaglutide microspheres prepared by the membrane emulsification method of the present invention have uniform particle size and adjustable size, have high drug loading and encapsulation efficiency, the organic solvent in the microspheres can be quickly removed with low residual content, are easy to scale up production, and the burst release of the microspheres is controllable, and can achieve 72 days of sustained release.

Claims

1. A method for preparing a semaglutide microsphere preparation, It is characterized in that The following steps are involved: (1) Add polyvinyl alcohol to water, heat to 70-80°C to dissolve, and cool to obtain a PVA solution; (2) adding the PEG copolymer to an organic solvent to dissolve, filtering, and adding semaglutide to the filtrate to dissolve to obtain a semaglutide solution; (3) Using semaglutide solution as dispersed phase and PVA solution as mobile phase, semaglutide emulsion was obtained by membrane emulsification method; (4) The semaglutide emulsion is homogenized and emulsified, the emulsion is stirred, the microspheres are solidified, washed, and dried to obtain a semaglutide microsphere preparation.

2. The method for preparing the semaglutide microsphere preparation according to claim 1, It is characterized in that The concentration of the PVA solution in step (1) is 0.2-5%; the weight average molecular weight of the polyvinyl alcohol is 84,000-124,000.

3. The method for preparing the semaglutide microsphere preparation according to claim 1, It is characterized in that The PEG copolymer in step (2) is one of PLGA-PEG-PLGA triblock copolymer, PEG-PLLA diblock copolymer, mPEG-PLGA diblock copolymer, and mPEG-PLLA diblock copolymer; the organic solvent is one of ethyl acetate, dichloromethane, chloroform, and tetrahydrofuran.

4. The method for preparing the semaglutide microsphere preparation according to claim 3, It is characterized in that The molar ratio of lactide to glycolide in the PLGA-PEG-PLGA triblock copolymer and the mPEG-PLGA diblock copolymer is 50-90:50-10; the molecular weight of PEG and mPEG in the PEG copolymer is 400-2000.

5. The method for preparing the semaglutide microsphere preparation according to claim 3, It is characterized in that The molar ratio of lactide to glycolide in the PLGA-PEG-PLGA triblock copolymer and the mPEG-PLGA diblock copolymer is 75-85:25-15; the molecular weight of mPEG in the mPEG-PLGA diblock copolymer and the mPEG-PLLA diblock copolymer is 800-1000.

6. The method for preparing the semaglutide microsphere preparation according to claim 1, It is characterized in that In step (2), the mass ratio of PEG copolymer to semaglutide is 1:0.5-2; the amount of organic solvent used is 8-10 times the mass of PEG copolymer.

7. The method for preparing the semaglutide microsphere preparation according to claim 1, It is characterized in that In step (3), the semaglutide solution is poured into a membrane emulsification tank, and nitrogen is passed through the tank to pressurize the tank to 0.05-0.25 MPa. The dispersed phase is extruded through the membrane tube, and the dispersed phase forms spherical droplets on the membrane surface. Under the action of the fast-flowing PVA solution, the droplets slowly detach from the membrane surface until no droplets are formed on the membrane surface. The nitrogen is then turned off to generate a semaglutide emulsion. In step (4), the drying method is vacuum freeze drying.

8. The method for preparing the semaglutide microsphere preparation according to claim 7, It is characterized in that The membrane tube described in step (3) is made of SPG membrane or stainless steel, and the pore size of the membrane tube is 1 to 200 μm.

9. The method for preparing the semaglutide microsphere preparation according to claim 1, It is characterized in that The homogenization emulsification conditions in step (4) are 15000~20000rpm, 20~40min.

10. A semaglutide microsphere preparation prepared by the preparation method according to any one of claims 1 to 9.

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