A liposome-encapsulated retalipotide formulation, methods of making and using the same

Abstract

The present application relates to the technical field of Retaliputide preparation, in particular to a liposome-encapsulated Retaliputide preparation, a preparation method and application thereof.The liposome-encapsulated Retaliputide preparation provided by the present application comprises Retaliputide and liposomes coated on the surface of the Retaliputide; the raw materials of the liposomes comprise hydrogenated lecithin, lecithin, cholesterol, o-sulfobenzamide and vitamin.The Retaliputide is encapsulated by the liposomes, so that it can be orally administered, the degradation of enzymes in the gastrointestinal tract can be largely avoided during oral administration, the bioavailability is ensured, and the use is convenient.Therefore, the liposome-encapsulated Retaliputide preparation provided by the present application has high bioavailability, convenient use, good social and economic benefits, and good application prospect.

Description

Technical Field

[0001] This invention relates to the field of retaglutide formulation technology, and in particular to a liposome-encapsulated retaglutide formulation, its preparation method, and its application. Background Technology

[0002] Retaglutide, a multi-target GLP-1 receptor agonist, aims to provide comprehensive treatment for obesity and type 2 diabetes by simultaneously activating the glucose-dependent insulinotropic peptide receptor (GIPR), glucagon-like peptide-1 receptor (GLP-1R), and glucagon receptor (GCGR). The drug's clinical trial results were announced in June 2023, and related research papers have been published in several authoritative international medical journals, demonstrating its outstanding therapeutic potential and broad application prospects.

[0003] Retaglutide is currently formulated as a subcutaneous injection solution, typically provided in pre-filled syringes (pre-filled syringes) for easy self-injection by patients. However, subcutaneous injection also has some drawbacks, potentially affecting patient compliance, especially for those who prefer oral medication.

[0004] Therefore, developing a more convenient formulation of retaglutide for patients is an urgent problem to be solved in this field, and it is of great significance for patients with obesity and type 2 diabetes. Summary of the Invention

[0005] In view of this, the present invention provides a liposome-encapsulated retaglutide formulation, its preparation method and application. The liposome-encapsulated retaglutide formulation provided by the present invention can be administered orally and is convenient to use.

[0006] This invention provides a liposome-encapsulated retaglutide formulation, comprising retaglutide and liposomes coated on the surface of retaglutide; the raw materials of the liposomes include hydrogenated lecithin, lecithin, cholesterol, o-sulfonylbenzoimide and vitamins.

[0007] This invention also provides a method for preparing the liposome-encapsulated retalotropic peptide formulation described above, comprising the following steps: (1) Hydrogenated lecithin, lecithin, cholesterol, vitamins and ethanol were mixed and then subjected to shearing, homogenization and solvent removal to obtain liposome membrane material; (2) The liposome membrane material was mixed with surfactant and water and subjected to hydration, shearing, purification and membrane rupture in sequence to obtain liposomes; (3) The liposomes were mixed with retaliglutide, o-sulfonylbenzoimide and water, encapsulated and filtered to obtain the liposome-encapsulated retaliglutide formulation.

[0008] Preferably, the hydrogenated lecithin is hydrogenated soybean lecithin; the lecithin is soybean lecithin.

[0009] Preferably, the mass ratio of hydrogenated lecithin to lecithin is 10:20~27; and the mass ratio of lecithin to cholesterol is 100:35~40.

[0010] Preferably, the vitamins include vitamin D and vitamin E; the mass ratio of vitamin D to vitamin E is 4~6:2~3; and the mass ratio of lecithin to vitamins is 100:1~3.

[0011] Preferably, the shearing speed is 12,000~15,000 rpm and the time is 5~10 minutes.

[0012] Preferably, the mass ratio of liposomes to retaliglutide is 1 to 2:1; the mass ratio of liposomes to o-sulfonylbenzoimide can be 100:35 to 45.

[0013] Preferably, the coating is carried out under the conditions of stirring and ultrasound; the ultrasound power is 80~100W; the stirring speed is 50~100rpm; and the coating time is 1~10 minutes.

[0014] Preferably, the filtration process further includes drying; the drying is freeze drying; the freeze drying includes pre-cooling, first freeze drying and second freeze drying in sequence; the pre-cooling method is: uniformly cooling from 4 degrees Celsius to -45 degrees Celsius for 60 to 120 minutes; the first freeze drying temperature is -45 degrees Celsius for 2 to 4 hours; the second freeze drying temperature is 30 degrees Celsius for 2 to 4 hours.

[0015] This invention also provides the application of the above-described liposome-encapsulated retaglutide formulation in the field of retaglutide drug preparation.

[0016] Compared with the prior art, the present invention has achieved the following beneficial effects: This invention encapsulates retaglutide in liposomes, enabling oral administration. This largely avoids enzymatic degradation in the gastrointestinal tract during oral administration, ensuring bioavailability and ease of use. Therefore, the liposome-encapsulated retaglutide formulation provided by this invention offers both high bioavailability and ease of use, demonstrating significant social and economic benefits and promising application prospects. This invention initially constructs liposomes from hydrogenated lecithin, lecithin, cholesterol, and vitamins, then hydrates them using a surfactant, and subsequently modifies the liposomes with o-sulfonylbenzoimide. This allows for the formulation of retaglutide in liposomes with good stability. Attached Figure Description

[0017] To more clearly illustrate the technical solutions of this invention, the accompanying drawings used in the embodiments of this invention or in the prior art are briefly described below. For those skilled in the art, other drawings can be derived from the following drawings without creative effort, and all such drawings are within the protection scope of this invention.

[0018] Figure 1 This is a schematic diagram of the structure of the liposome-encapsulated retaglutide formulation provided by the present invention; wherein, 1 is retaglutide and 2 is a liposome; Figure 2 This is a scanning electron microscope image of the liposome-encapsulated retalotropic peptide preparation prepared in Example 1 of the present invention. Detailed Implementation

[0019] This invention provides a liposome-encapsulated retaglutide formulation, comprising retaglutide and liposomes coated on the surface of retaglutide; the raw materials of the liposomes include hydrogenated lecithin, lecithin, cholesterol, o-sulfonylbenzoimide and vitamins.

[0020] This invention also provides a method for preparing the liposome-encapsulated retalotropic peptide formulation described above, comprising the following steps: (1) Hydrogenated lecithin, lecithin, cholesterol, vitamins and ethanol were mixed and then subjected to shearing, homogenization and solvent removal to obtain liposome membrane material; (2) The liposome membrane material was mixed with surfactant and water and subjected to hydration, shearing, purification and membrane rupture in sequence to obtain liposomes; (3) The liposomes were mixed with retaliglutide, o-sulfonylbenzoimide and water, encapsulated and filtered to obtain the liposome-encapsulated retaliglutide formulation.

[0021] This invention involves mixing hydrogenated lecithin, lecithin, cholesterol, vitamins, and ethanol, followed by sequential shearing, homogenization, and solvent removal to obtain a liposome membrane material. In this invention, the hydrogenated lecithin can be hydrogenated soybean lecithin.

[0022] In this invention, lecithin can be soybean lecithin.

[0023] In this invention, the mass ratio of hydrogenated lecithin to lecithin can be 10:20~27, specifically 10:21, 10:22, 10:23, 10:24, 10:25 or 10:26.

[0024] In this invention, the mass ratio of lecithin to cholesterol can be 100:35~40, specifically 100:36, 100:37, 100:38 or 100:39.

[0025] In this invention, vitamins may include vitamin D and vitamin E; the mass ratio of vitamin D to vitamin E may be 4~6:2~3, specifically 5:2.4 or 5:2.7.

[0026] In this invention, the mass ratio of lecithin to vitamins can be 100:1 to 3, specifically 100:2.

[0027] In this invention, ethanol can be anhydrous ethanol.

[0028] In this invention, the mass ratio of lecithin to ethanol can be 1:600~800, specifically 1:650, 1:700 or 1:750.

[0029] In this invention, the shearing speed can be 12,000 to 15,000 rpm, specifically 13,000 rpm or 14,000 rpm, and the time can be 5 to 10 minutes, specifically 8 minutes.

[0030] In this invention, the homogenization pressure can be 14~16MPa, specifically 15MPa, and the time can be 5~10 minutes, specifically 7 minutes.

[0031] In this invention, the solvent can be evaporation, the evaporation temperature can be 60~70 degrees Celsius, specifically 65 degrees Celsius, the vacuum degree can be -0.8~-0.95 MPa, specifically -0.9 MPa, and the time can be more than 30 minutes, specifically 60 minutes, 120 minutes or 180 minutes.

[0032] After obtaining the liposome membrane material, this invention mixes the liposome membrane material with a surfactant and water, and sequentially performs hydration, shearing, purification, and membrane disruption to obtain liposomes. In this invention, the surfactant can be Tween 80.

[0033] In this invention, the mass ratio of liposome membrane material to surfactant can be 13 to 17:1, specifically 14:1, 15:1 or 16:1.

[0034] In this invention, the mass ratio of surfactant to water can be 1:120 to 160, specifically 1:130, 1:140 or 1:150.

[0035] In this invention, the hydration temperature can be 55-60 degrees Celsius, specifically 58 degrees Celsius, and the time can be 1-3 hours, specifically 1.5 hours or 2 hours.

[0036] In this invention, the shearing speed can be 12,000 to 15,000 rpm, specifically 13,000 rpm or 14,000 rpm, and the time can be 5 to 10 minutes, specifically 8 minutes.

[0037] In this invention, purification can be ultrafiltration; the parameters of ultrafiltration can include: a 100kDa ultrafiltration membrane or a 300kDa ultrafiltration membrane, and a time of 25 to 35 minutes, specifically 30 minutes.

[0038] After obtaining the liposomes, the present invention mixes the liposomes with retaglutide, o-sulfonylbenzoimide and water, encapsulates the mixture, and then filters it to obtain a liposome-encapsulated retaglutide formulation. In the present invention, the mass ratio of liposomes to retaglutide can be 1~2:1, specifically 1.3:1, 1.5:1 or 1.8:1.

[0039] In this invention, the mass ratio of liposomes to o-sulfonylbenzoimide can be 100:35~45, specifically 100:37, 100:39, 100:40 or 100:43.

[0040] In this invention, the mass ratio of liposomes to water can be 1:50~70, specifically 1:55, 1:60 or 1:65.

[0041] In this invention, the wrapping can be carried out under stirring and ultrasonic conditions; the ultrasonic power can be 80~100W, specifically 90W; the stirring speed can be 50~100rpm, specifically 60rpm, 70rpm, 80rpm or 90rpm; the wrapping time can be 1~10 minutes, specifically 3 minutes, 5 minutes or 8 minutes.

[0042] In this invention, the filtration process may further include drying; the drying process may be freeze drying; the freeze drying process may include pre-cooling, first freeze drying, and second freeze drying in sequence; the pre-cooling method may be: uniformly cooling from 4 degrees Celsius to -45 degrees Celsius for 60 to 120 minutes, specifically 75 minutes or 90 minutes; the first freeze drying temperature may be -45 degrees Celsius for 2 to 4 hours, specifically 3 hours; the second freeze drying temperature may be 30 degrees Celsius for 2 to 4 hours, specifically 3 hours.

[0043] This invention also provides the application of the above-described liposome-encapsulated retaglutide formulation in the field of retaglutide drug preparation.

[0044] The liposome-encapsulated retalotropic peptide formulation provided by this invention has the following structure: Figure 1 As shown, it has the advantages of good stability, high bioavailability and oral administration, making it suitable for the preparation of retaglutide drugs, with broad application prospects.

[0045] To further illustrate the present invention, the technical solutions of the present invention will be clearly and completely described below with reference to the accompanying drawings and embodiments.

[0046] Example 1: This embodiment prepares a liposome-encapsulated retaglutide formulation, and the specific steps are as follows: (1) Mix 10g hydrogenated soybean lecithin, 25g soybean lecithin, 9.2g cholesterol, 0.5g vitamin D, 0.25g vitamin E and 18kg anhydrous ethanol, shear at 15000rpm for 8 minutes, homogenize at 14MPa for 7 minutes, and evaporate at 65 degrees Celsius and -0.9MPa for 30 minutes to remove solvent to obtain liposome membrane material.

[0047] (2) Take 15g of the obtained liposome membrane material, mix the liposome membrane material with 1g Tween 80 and 150g of water, hydrate at 58 degrees Celsius for 1.5 hours, shear at 13000rpm for 8 minutes, purify with a 100kDa ultrafiltration membrane for 30 minutes, break the membrane, and obtain liposomes.

[0048] (3) Mix 5g of the prepared liposomes with 2.5g of retaliglutide, 2g of o-sulfonylbenzoimide and 300g of water, and encapsulate them under stirring and sonication. The sonication power is 90W and the stirring speed is 80rpm. Encapsulate for 3 minutes, filter, cool from 4 degrees Celsius to -45 degrees Celsius at a constant speed for 90 minutes, freeze-dry at -45 degrees Celsius for 3 hours, and freeze-dry at 30 degrees Celsius for 3 hours to obtain the liposome-encapsulated retaliglutide formulation.

[0049] Example 2: This embodiment prepares a liposome-encapsulated retaglutide formulation, and the specific steps are as follows: (1) Mix 10g hydrogenated soybean lecithin, 25g soybean lecithin, 9.2g cholesterol, 0.5g vitamin D, 0.25g vitamin E and 18kg anhydrous ethanol, shear at 15000rpm for 8 minutes, homogenize at 14MPa for 7 minutes, and evaporate at 65 degrees Celsius and -0.9MPa for 30 minutes to remove solvent to obtain liposome membrane material.

[0050] (2) Take 15g of the obtained liposome membrane material, mix the liposome membrane material with 1g Tween 80 and 150g of water, hydrate at 58 degrees Celsius for 1.5 hours, shear at 13000rpm for 8 minutes, purify with a 100kDa ultrafiltration membrane for 30 minutes, break the membrane, and obtain liposomes.

[0051] (3) Mix 5g of the prepared liposomes with 3g of retaliglutide, 2g of o-sulfonylbenzoimide and 300g of water, and encapsulate them under stirring and sonication. The sonication power is 80W and the stirring speed is 80rpm. Encapsulate for 3 minutes, filter, cool from 4 degrees Celsius to -45 degrees Celsius at a uniform rate for 90 minutes, freeze-dry at -45 degrees Celsius for 3 hours, and freeze-dry at 30 degrees Celsius for 3 hours to obtain the liposome-encapsulated retaliglutide formulation.

[0052] Example 3: This embodiment prepares a liposome-encapsulated retaglutide formulation, and the specific steps are as follows: (1) Mix 10g hydrogenated soybean lecithin, 25g soybean lecithin, 9.2g cholesterol, 0.5g vitamin D, 0.25g vitamin E and 18kg anhydrous ethanol, shear at 15000rpm for 8 minutes, homogenize at 14MPa for 7 minutes, and evaporate at 65 degrees Celsius and -0.9MPa for 30 minutes to remove solvent to obtain liposome membrane material.

[0053] (2) Take 15g of the obtained liposome membrane material, mix the liposome membrane material with 1g Tween 80 and 150g of water, hydrate at 58 degrees Celsius for 1.5 hours, shear at 13000rpm for 8 minutes, purify with a 100kDa ultrafiltration membrane for 30 minutes, break the membrane, and obtain liposomes.

[0054] (3) Mix 5g of the prepared liposomes with 3.5g of retaliglutide, 2g of o-sulfonylbenzoimide and 300g of water, and encapsulate them under stirring and sonication. The sonication power is 80W and the stirring speed is 80rpm. Encapsulate for 5 minutes, filter, cool from 4 degrees Celsius to -45 degrees Celsius at a constant speed for 90 minutes, freeze-dry at -45 degrees Celsius for 3 hours, and freeze-dry at 30 degrees Celsius for 3 hours to obtain the liposome-encapsulated retaliglutide formulation.

[0055] Example 4: This embodiment prepares a liposome-encapsulated retaglutide formulation, and the specific steps are as follows: (1) Mix 10g hydrogenated soybean lecithin, 25g soybean lecithin, 9.2g cholesterol, 0.5g vitamin D, 0.25g vitamin E and 18kg anhydrous ethanol, shear at 15000rpm for 8 minutes, homogenize at 14MPa for 7 minutes, and evaporate at 65 degrees Celsius and -0.9MPa for 30 minutes to remove solvent to obtain liposome membrane material.

[0056] (2) Take 15g of the obtained liposome membrane material, mix the liposome membrane material with 1g Tween 80 and 150g of water, hydrate at 58 degrees Celsius for 1.5 hours, shear at 13000rpm for 8 minutes, purify with a 100kDa ultrafiltration membrane for 30 minutes, break the membrane, and obtain liposomes.

[0057] (3) Mix 5g of the prepared liposomes with 4g of retaliglutide, 2.1g of o-sulfonylbenzoimide and 300g of water, and encapsulate them under stirring and sonication. The sonication power is 90W and the stirring speed is 80rpm. Encapsulate for 5 minutes, filter, cool from 4 degrees Celsius to -45 degrees Celsius at a uniform rate for 90 minutes, freeze-dry at -45 degrees Celsius for 3 hours, and freeze-dry at 30 degrees Celsius for 3 hours to obtain the liposome-encapsulated retaliglutide formulation.

[0058] Example 5: This embodiment prepares a liposome-encapsulated retaglutide formulation, and the specific steps are as follows: (1) Mix 10g hydrogenated soybean lecithin, 25g soybean lecithin, 9.2g cholesterol, 0.5g vitamin D, 0.25g vitamin E and 18kg anhydrous ethanol, shear at 15000rpm for 8 minutes, homogenize at 14MPa for 7 minutes, and evaporate at 65 degrees Celsius and -0.9MPa for 30 minutes to remove solvent to obtain liposome membrane material.

[0059] (2) Take 15g of the obtained liposome membrane material, mix the liposome membrane material with 1g Tween 80 and 150g of water, hydrate at 58 degrees Celsius for 1.5 hours, shear at 13000rpm for 8 minutes, purify with a 100kDa ultrafiltration membrane for 30 minutes, break the membrane, and obtain liposomes.

[0060] (3) Mix 5g of the prepared liposomes with 4g of retaliglutide, 2g of o-sulfonylbenzoimide and 300g of water, and encapsulate them under stirring and sonication. The sonication power is 100W and the stirring speed is 80rpm. Encapsulate for 5 minutes, filter, cool from 4 degrees Celsius to -45 degrees Celsius at a constant speed for 80 minutes, freeze-dry at -45 degrees Celsius for 3 hours, and freeze-dry at 30 degrees Celsius for 3 hours to obtain the liposome-encapsulated retaliglutide formulation.

[0061] Example 6: This embodiment prepares a liposome-encapsulated retaglutide formulation, and the specific steps are as follows: (1) Mix 10g hydrogenated soybean lecithin, 25g soybean lecithin, 9.2g cholesterol, 0.5g vitamin D, 0.25g vitamin E and 18kg anhydrous ethanol, shear at 15000rpm for 8 minutes, homogenize at 14MPa for 7 minutes, and evaporate at 65 degrees Celsius and -0.9MPa for 30 minutes to remove solvent to obtain liposome membrane material.

[0062] (2) Take 15g of the obtained liposome membrane material, mix the liposome membrane material with 1g Tween 80 and 150g of water, hydrate at 58 degrees Celsius for 1.5 hours, shear at 13000rpm for 8 minutes, purify with a 100kDa ultrafiltration membrane for 30 minutes, break the membrane, and obtain liposomes.

[0063] (3) Mix 5g of the prepared liposomes with 4.5g of retaliglutide, 2.1g of o-sulfonylbenzoimide and 300g of water, and encapsulate them under stirring and sonication. The sonication power is 100W and the stirring speed is 80rpm. After encapsulation for 5 minutes, filter, cool from 4 degrees Celsius to -45 degrees Celsius at a uniform rate for 95 minutes, freeze-dry at -45 degrees Celsius for 3 hours, and freeze-dry at 30 degrees Celsius for 3 hours to obtain the liposome-encapsulated retaliglutide formulation.

[0064] Example 7: This embodiment prepares a liposome-encapsulated retaglutide formulation, and the specific steps are as follows: (1) Mix 10g hydrogenated soybean lecithin, 25g soybean lecithin, 9.2g cholesterol, 0.5g vitamin D, 0.25g vitamin E and 18kg anhydrous ethanol, shear at 15000rpm for 8 minutes, homogenize at 14MPa for 7 minutes, and evaporate at 65 degrees Celsius and -0.9MPa for 30 minutes to remove solvent to obtain liposome membrane material.

[0065] (2) Take 15g of the obtained liposome membrane material, mix the liposome membrane material with 1g Tween 80 and 150g of water, hydrate at 58 degrees Celsius for 1.5 hours, shear at 13000rpm for 8 minutes, purify with a 100kDa ultrafiltration membrane for 30 minutes, break the membrane, and obtain liposomes.

[0066] (3) Mix 5g of the prepared liposomes with 4.5g of retaliglutide, 2g of o-sulfonylbenzoimide and 300g of water, and encapsulate them under stirring and sonication. The sonication power is 90W and the stirring speed is 80rpm. Encapsulate for 1 minute, filter, cool from 4 degrees Celsius to -45 degrees Celsius at a uniform rate for 100 minutes, freeze-dry at -45 degrees Celsius for 3 hours, and freeze-dry at 30 degrees Celsius for 3 hours to obtain the liposome-encapsulated retaliglutide formulation.

[0067] Example 8: This embodiment prepares a liposome-encapsulated retaglutide formulation, and the specific steps are as follows: (1) Mix 10g hydrogenated soybean lecithin, 25g soybean lecithin, 9.2g cholesterol, 0.5g vitamin D, 0.25g vitamin E and 18kg anhydrous ethanol, shear at 15000rpm for 8 minutes, homogenize at 14MPa for 7 minutes, and evaporate at 65 degrees Celsius and -0.9MPa for 30 minutes to remove solvent to obtain liposome membrane material.

[0068] (2) Take 15g of the obtained liposome membrane material, mix the liposome membrane material with 1g Tween 80 and 150g of water, hydrate at 58 degrees Celsius for 1.5 hours, shear at 13000rpm for 8 minutes, purify with a 100kDa ultrafiltration membrane for 30 minutes, break the membrane, and obtain liposomes.

[0069] (3) Mix 5g of the prepared liposomes with 5g of retaliglutide, 2g of o-sulfonylbenzoimide and 300g of water, and encapsulate them under stirring and sonication. The sonication power is 80W and the stirring speed is 80rpm. Encapsulate for 1 minute, filter, cool from 4 degrees Celsius to -45 degrees Celsius at a constant speed for 95 minutes, freeze-dry at -45 degrees Celsius for 3 hours, and freeze-dry at 30 degrees Celsius for 3 hours to obtain the liposome-encapsulated retaliglutide formulation.

[0070] Example 9: This embodiment prepares a liposome-encapsulated retaglutide formulation, and the specific steps are as follows: (1) Mix 10g hydrogenated soybean lecithin, 25g soybean lecithin, 9.2g cholesterol, 0.5g vitamin D, 0.25g vitamin E and 18kg anhydrous ethanol, shear at 15000rpm for 8 minutes, homogenize at 14MPa for 7 minutes, and evaporate at 65 degrees Celsius and -0.9MPa for 30 minutes to remove solvent to obtain liposome membrane material.

[0071] (2) Take 15g of the obtained liposome membrane material, mix the liposome membrane material with 1g Tween 80 and 150g of water, hydrate at 58 degrees Celsius for 1.5 hours, shear at 13000rpm for 8 minutes, purify with a 100kDa ultrafiltration membrane for 30 minutes, break the membrane, and obtain liposomes.

[0072] (3) Mix 5g of the prepared liposomes with 5g of retaliglutide, 2.1g of o-sulfonylbenzoimide and 300g of water, and encapsulate them under stirring and sonication. The sonication power is 100W and the stirring speed is 80rpm. Encapsulate for 3 minutes, filter, cool from 4 degrees Celsius to -45 degrees Celsius at a constant speed for 90 minutes, freeze-dry at -45 degrees Celsius for 3 hours, and freeze-dry at 30 degrees Celsius for 3 hours to obtain the liposome-encapsulated retaliglutide formulation.

[0073] Example 10: This embodiment prepares a liposome-encapsulated retaglutide formulation, and the specific steps are as follows: (1) Mix 10g hydrogenated soybean lecithin, 25g soybean lecithin, 9.2g cholesterol, 0.5g vitamin D, 0.25g vitamin E and 18kg anhydrous ethanol, shear at 15000rpm for 8 minutes, homogenize at 14MPa for 7 minutes, and evaporate at 65 degrees Celsius and -0.9MPa for 30 minutes to remove solvent to obtain liposome membrane material.

[0074] (2) Take 15g of the obtained liposome membrane material, mix the liposome membrane material with 1g Tween 80 and 150g of water, hydrate at 58 degrees Celsius for 1.5 hours, shear at 13000rpm for 8 minutes, purify with a 100kDa ultrafiltration membrane for 30 minutes, break the membrane, and obtain liposomes.

[0075] (3) Mix 5g of the prepared liposomes with 5g of retaliglutide, 2g of o-sulfonylbenzoimide and 300g of water, and encapsulate them under stirring and sonication. The sonication power is 90W and the stirring speed is 80rpm. Encapsulate for 3 minutes, filter, cool from 4 degrees Celsius to -45 degrees Celsius at a constant speed for 90 minutes, freeze-dry at -45 degrees Celsius for 3 hours, and freeze-dry at 30 degrees Celsius for 3 hours to obtain the liposome-encapsulated retaliglutide formulation.

[0076] Comparative Example 1: The preparation method of this comparative example is the same as that of Example 1, except that hydrogenated soybean lecithin is replaced with an equal mass of soybean lecithin.

[0077] Comparative Example 2: The preparation method of this comparative example is the same as that of Example 5, except that soybean lecithin is replaced with an equal mass of hydrogenated soybean lecithin.

[0078] Comparative Example 3: The preparation method of this comparative example is the same as that of Example 3, except that vitamin D is replaced with an equal mass of vitamin E.

[0079] Comparative Example 4: The preparation method of this comparative example is the same as that of Example 1, except that the mass of Retaglutide is 2g.

[0080] Comparative Example 5: The preparation method of this comparative example is the same as that of Example 1, except that the mass of Retaglutide is 4g.

[0081] Comparative Example 6: The preparation method of this comparative example is the same as that of Example 1, except that o-sulfonylbenzoimide was not added.

[0082] Test Example 1: The liposome-encapsulated retalotropic peptide preparation prepared in Example 1 was characterized using scanning electron microscopy, and the results are as follows: Figure 2 As shown.

[0083] according to Figure 2 As can be seen, the liposome-encapsulated retaliglutide formulation provided by the present invention is structurally a liposome-encapsulated retaliglutide, which proves that the present invention has successfully encapsulated retaliglutide in liposomes.

[0084] Test Example 2: The performance of the liposome-encapsulated retaglutide formulations of Examples 1-10 and Comparative Examples 1-6 was tested. The test method was as follows: 380 male SD (Sprague-Dawley) rats aged 6-8 weeks were selected, of which 360 were male SD rats with type II diabetes model (induced by high-fat / high-sugar diet combined with low-dose streptozotocin), and 20 were healthy male SD rats. Apart from whether or not the model was established, there were no other differences in the physiological health status and subsequent feeding process of the above rats. 360 rats were randomly divided into 18 groups. Each group was given either the liposome-encapsulated retalistatin formulation of Examples 1-10 or Comparative Examples 1-6, an aqueous solution of an equal volume of retalistatin powder (commercial control group), or an equal volume of physiological saline (model control group). 20 healthy male SD rats served as the blank control group (receiving an equal volume of physiological saline). Each rat was given the drug (liposome-encapsulated retalistatin formulation, aqueous solution of retalistatin powder, or physiological saline) once a week according to its group. The test was conducted for 60 weeks. Fasting blood glucose and 2-hour postprandial blood glucose on the first day of each week were recorded for all rats in each group, and the average values ​​were calculated. The results are shown in Tables 1 and 2.

[0085] Table 1. Fasting blood glucose and physiological test results of Examples 1-10 and Comparative Examples 1-6:

[0086] Table 2. Postprandial blood glucose test results of Examples 1-10 and Comparative Examples 1-6:

[0087] As shown in Tables 1 and 2, the liposome-encapsulated retaglutide formulation provided by this invention effectively protects retaglutide from degradation by gastrointestinal enzymes, ensuring that it can reach its site of action and exert its effect. Compared to commercially available retaglutide powder, the liposome-encapsulated retaglutide formulation provided by this invention can better control blood glucose levels under the same conditions, demonstrating a significant improvement in bioavailability. This provides a new approach to reducing medication costs for patients and ensuring therapeutic efficacy.

[0088] The embodiments of the present invention have been described above; however, these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. All other embodiments obtained by those skilled in the art based on the above embodiments of the present invention without inventive effort are within the protection scope of the present invention.

Claims

1. A liposome-encapsulated retalotropic peptide formulation, characterized in that, It includes retaliglutide and liposomes coated on the surface of retaliglutide; the raw materials of the liposomes include hydrogenated lecithin, lecithin, cholesterol, o-sulfonylbenzoimide and vitamins.

2. A method for preparing a liposome-encapsulated retalotropic peptide formulation, characterized in that, The liposome-encapsulated retaliglutide formulation of claim 1 comprises the following steps: (1) Hydrogenated lecithin, lecithin, cholesterol, vitamins and ethanol were mixed and then subjected to shearing, homogenization and solvent removal to obtain liposome membrane material; (2) The liposome membrane material was mixed with surfactant and water and subjected to hydration, shearing, purification and membrane rupture in sequence to obtain liposomes; (3) The liposomes were mixed with retaliglutide, o-sulfonylbenzoimide and water, encapsulated and filtered to obtain the liposome-encapsulated retaliglutide formulation.

3. The preparation method according to claim 2, characterized in that, Hydrogenated lecithin is hydrogenated soybean lecithin; lecithin is soybean lecithin.

4. The preparation method according to claim 2, characterized in that, The mass ratio of hydrogenated lecithin to lecithin is 10:20~27; the mass ratio of lecithin to cholesterol is 100:35~40.

5. The preparation method according to claim 2, characterized in that, Vitamins include vitamin D and vitamin E; the mass ratio of vitamin D to vitamin E is 4~6:2~3; the mass ratio of lecithin to vitamins is 100:1~3.

6. The preparation method according to claim 2, characterized in that, In step (1), the shearing speed is 12000~15000rpm and the time is 5~10 minutes.

7. The preparation method according to claim 2, characterized in that, The mass ratio of liposomes to retaliglutide is 1~2:1; the mass ratio of liposomes to o-sulfonylbenzoimide can be 100:35~45.

8. The preparation method according to claim 2, characterized in that, The coating process is carried out under stirring and ultrasonic conditions; the ultrasonic power is 80~100W; the stirring speed is 50~100rpm; and the coating time is 1~10 minutes.

9. The preparation method according to claim 2, characterized in that, After filtration, drying is also included; the drying is freeze drying; the freeze drying includes pre-cooling, first freeze drying and second freeze drying in sequence; the pre-cooling method is: uniformly cooling from 4 degrees Celsius to -45 degrees Celsius for 60 to 120 minutes; the temperature of the first freeze drying is -45 degrees Celsius for 2 to 4 hours; the temperature of the second freeze drying is 30 degrees Celsius for 2 to 4 hours.

10. The application of a liposome-encapsulated retaglutide formulation in the field of retaglutide drug preparation, characterized in that, The liposome-encapsulated retaliglutide formulation is the liposome-encapsulated retaliglutide formulation of claim 1.

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