Smeglutide self-emulsifying composition and application thereof

Abstract

The invention discloses a semeglutide self-emulsifying composition and application thereof, and belongs to the technical field of biological medicine. According to the invention, a hydrophobic ion pairing (HIP) technology is utilized, the semeglutide and counter ions are firstly prepared into a hydrophobic ion pair compound to improve the lipid solubility of the semeglutide, and then the semeglutide hydrophobic ion pair compound is loaded into the self-microemulsion as an active component to realize efficient oral delivery of the semeglutide. Based on a hydrophobic ion pairing technology and a self-microemulsion delivery technology, a novel simple and cost-effective semeglutide oral delivery method is constructed, and the method has a good application prospect.

Description

Technical Field

[0001] The present invention belongs to the technical field of biomedicine, and particularly relates to a semaglutide self-emulsifying composition and application thereof. Background Art

[0002] Diabetes is a chronic disease that causes metabolic disorders primarily manifesting as abnormal metabolism of carbohydrates, fats, and proteins. This disorder can be caused by relative or absolute insulin deficiency, structural defects in the insulin itself, or decreased insulin sensitivity in target cells. The primary clinical manifestations are hyperglycemia and glycosuria, with symptoms of "three mores and one less." Based on its pathogenesis, diabetes can be divided into four types: type 1, type 2, specific type, and gestational type. Type 2 diabetes is the most common type of diabetes, accounting for 90% to 95% of all patients. Therefore, the search for effective treatments and medications for type 2 diabetes is urgent.

[0003] Glucagon-like peptide-1 (GLP-1) is a glucose-dependent insulinotropic hormone that increases insulin secretion. This effect occurs only in response to elevated blood glucose levels, not at normal blood glucose levels. Furthermore, GLP-1 can slow β-cell apoptosis and promote β-cell regeneration, offering a new treatment option for type 2 diabetes. However, its short half-life in vivo, rapid degradation by dipeptidyl peptidase-IV (DPP-IV), and renal metabolism significantly limit its clinical application. Consequently, researchers have been diligently developing new, more stable GLP-1 analogs to resist degradation and extend their biological half-life. These drugs mimic the effects of native GLP-1 to activate the GLP-1 receptor, thereby enhancing insulin secretion, inhibiting glucagon secretion, and delaying gastric emptying in a glucose-dependent manner. These drugs not only lower blood glucose but also have other benefits, including weight loss, blood pressure reduction, and improved lipid profiles.

[0004] GLP-1 analogs, as a class of polypeptide drugs, can currently only be administered by injection. In order to improve patient compliance, sustained-release injections and non-injection administration have become the main research and development directions for this type of drug. Among them, exenatide is made into sustained-release microspheres, which can be administered once a week. Some products directly avoid the disadvantages of injection and develop products such as implants, oral administration, transdermal and inhalation. Among them, oral administration is the most ideal route of administration, but it is necessary to solve many problems such as the destruction and absorption of GLP-1 analogs by gastric acid and enzymes in the stomach and intestines in order to improve bioavailability and reduce individual differences. Since diabetes is a chronic disease that requires long-term treatment, the existing GLP-1 analog preparations have a short duration of action and need to be injected daily or weekly. Long-term injections bring great pain and inconvenience to patients, resulting in poor patient compliance.

[0005] Semaglutide, a GLP-1 analog, is used to treat type 2 diabetes. Currently available formulations include subcutaneous injection and oral tablets. Semaglutide injection is primarily used to control blood sugar in adults with type 2 diabetes who, despite diet and exercise, have not yet achieved target blood sugar levels despite treatment with metformin or sulfonylureas. This subcutaneous injection utilizes a modified human GLP-1 molecule to achieve resistance to degradation by dipeptidyl peptidase 4 (DPP-4) and tight albumin binding, resulting in a longer half-life. It also retains high GLP-1 amino acid sequence homology, ensuring its safety and efficacy. Despite the significant advantages of GLP-1RAs in terms of glucose lowering and cardiovascular benefits, patient acceptance of GLP-1RA therapy remains low, primarily due to gastrointestinal adverse reactions and the subcutaneous route of administration. Subcutaneous injection not only reduces drug compliance but also is prone to eliciting an immune response. Therefore, Novo Nordisk obtained approval from the National Medical Products Administration (NMPA) and launched oral semaglutide tablets for the treatment of type 2 diabetes. This is the first oral GLP-1 receptor agonist approved for marketing in China. This tablet provides patients with a new treatment option and is easier for patients to use and manage. However, the bioavailability of semaglutide oral tablets is only 0.5-1.0%, resulting in an oral dose much larger than the injection dose. At the same time, in order to improve bioavailability, SNAC is used in the prescription of semaglutide oral tablets. SNAC is not selective for the passage of drugs through the small intestinal wall, so oral semaglutide requires fasting for 6 hours before taking it and fasting for half an hour after taking it, which causes inconvenience to the use of semaglutide oral tablets.

[0006] Due to the special properties of semaglutide (its large molecular weight makes it difficult to be absorbed through the gastrointestinal mucosa and it is degraded by enzymes and acids and bases in the gastrointestinal tract), the research on its oral formulation brings great difficulties and severe challenges, especially in terms of improving oral bioavailability. Overcoming the obstacles to oral delivery of semaglutide and improving its bioavailability are urgent problems to be solved. Based on this, the present invention designs and utilizes hydrophobic ion pairing (HIP) technology to increase the fat solubility of semaglutide, and then loads it into a self-microemulsion to achieve efficient oral delivery of semaglutide. Summary of the Invention

[0007] The purpose of the present invention is to provide a semaglutide self-emulsifying composition and its application, and to create a simple and cost-effective new method for oral delivery of semaglutide based on hydrophobic ion pairing technology and self-microemulsification delivery technology.

[0008] In order to achieve the above object, the present invention adopts the following technical solutions:

[0009] A semaglutide hydrophobic ion pair complex is generated by the reaction of semaglutide and a counterion, wherein the counterion is docusate sodium, sodium lauryl sulfate or sodium lauryl sulfonate; and the molar ratio of the semaglutide to the counterion is 1:1-6:1.

[0010] Furthermore, the counterion is docusate sodium, and the molar ratio of semaglutide to docusate sodium is 1:1-6:1, preferably 2:1-6:1, and more preferably 4:1.

[0011] Furthermore, the preparation method of the semaglutide hydrophobic ion pair complex is as follows: adding docusate sodium to water to obtain a docusate sodium aqueous solution, adding semaglutide to a hydrochloric acid solution to obtain a semaglutide hydrochloric acid solution, adding the docusate sodium aqueous solution to the semaglutide hydrochloric acid solution, stirring to react, centrifuging the reaction solution, washing the precipitate, and freeze-drying to obtain the semaglutide hydrophobic ion pair complex.

[0012] In a specific embodiment of the present invention, the method for preparing the semaglutide-docusate sodium ion pair comprises the following specific steps:

[0013] Step 1: Weigh docusate sodium, add an appropriate amount of ultrapure water to prepare a docusate sodium aqueous solution; Step 2: Weigh semaglutide, use 0.01 mol / L HCl solution as a solvent, and prepare a semaglutide hydrochloride solution; Step 3: Add docusate sodium to the semaglutide solution according to the molar ratio, stir at a speed of 500 rpm at room temperature for 30 minutes, and the solution becomes white turbid; Step 4: Centrifuge the suspension at 11220 rpm for 5 minutes, discard the supernatant, wash the precipitate with ultrapure water 2-3 times, collect the precipitate, and freeze-dry; Step 5: The final white powder is the semaglutide-docusate sodium ion pair, which is collected in a penicillin bottle and stored in a sealed dark place in a refrigerator at 4°C for later use.

[0014] A semaglutide self-emulsifying composition, the raw materials of which include an active ingredient and an excipient mixture; the active ingredient is the above-mentioned semaglutide hydrophobic ion pair complex, and the excipient mixture includes an oil phase, an emulsifier, and a cosolvent;

[0015] The amount of the semaglutide hydrophobic ion pair complex is 0.01-0.27% of the mass of the auxiliary material mixture.

[0016] Furthermore, the oil phase is a mixture of propylene glycol monocaprylate and medium chain triglycerides. Preferably, the mass ratio of propylene glycol monocaprylate to medium chain triglycerides is 2:1-1:3, more preferably 1:2.

[0017] Furthermore, the oil phase accounts for 10%-40% of the total mass of the auxiliary material mixture, preferably 10%-30%, more preferably 30%.

[0018] Furthermore, the emulsifier is caprylic acid capric acid polyethylene glycol glyceride, and the co-solvent is diethylene glycol monoethyl ether.

[0019] Furthermore, the mass ratio of the emulsifier to the co-solvent is 1:2-4:1, preferably 2.5:1.

[0020] In a specific embodiment of the present invention, the mass ratio of the oil phase, the emulsifier and the co-solvent in the auxiliary material mixture is 3:5:2.

[0021] A semaglutide self-microemulsification composition comprises the semaglutide self-emulsification composition and water.

[0022] In a specific embodiment of the present invention, the preparation method of the semaglutide-docusate sodium ion pair self-microemulsion is: weighing the prescribed amount of excipients and mixing them evenly, adding the prescribed amount of semaglutide-docusate sodium complex, mixing evenly, taking the mixture and adding 100 times the amount of 37°C ultrapure water, stirring to obtain a semaglutide-docusate sodium ion pair self-microemulsion.

[0023] Use of the above-mentioned semaglutide self-emulsifying composition or the above-mentioned semaglutide self-microemulsification in preparing a medicament, wherein the medicament is used for preventing and treating diabetes, obesity, non-alcoholic fatty liver disease or neurodegenerative diseases.

[0024] The present invention is based on hydrophobic ion pairing technology and self-microemulsion delivery technology, and uses a hydrophobic ion pair complex generated by the reaction of semaglutide with a counterion as an active ingredient to prepare a self-microemulsion.

[0025] Through the inventors' preliminary experiments, they selected docusate sodium as a counterion from commonly used counterion reagents. The sulfonate group in its chemical structure is strongly acidic and can form a hydrophobic ion pair with semaglutide. The branched structure is hydrophobic, and after forming an ion pair with semaglutide, it helps to improve the lipid solubility of semaglutide. Docusate sodium has the following structure:

[0026]

[0027] The present invention is based on hydrophobic ion pairing technology and self-microemulsification delivery technology to construct a simple and cost-effective new method for oral delivery of semaglutide, which has good application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 is the complexation rate of semaglutide and different counterions.

[0029] Figure 2 is the logP of the counterion complex formed by semaglutide and different counterions.

[0030] Figure 3 is the DSC spectrum of SET-DOC complex.

[0031] Figure 4 、 Figure 5 、 Figure 6 is the equilibrium solubility of SET-DOC complex in different oily lysozymes.

[0032] Figure 7 It is a pseudo-ternary phase diagram.

[0033] Figure 8 This is the screening result of oil phase ratio.

[0034] Figure 9 This is the screening result of the ratio of emulsifier and co-emulsifier.

[0035] Figure 10 The results are for the in vitro release of semaglutide-docusate sodium ion pair from microemulsion.

[0036] Figure 11 The results are for the cytotoxicity of semaglutide-docusate sodium ions on self-microemulsion.

[0037] Figure 12 These are the pharmacodynamic results of semaglutide-docusate sodium ion pair self-microemulsion. DETAILED DESCRIPTION

[0038] The preferred embodiments of the present invention will be described in detail below with reference to the examples. It should be understood that the following examples are provided for illustrative purposes only and are not intended to limit the scope of the present invention. Those skilled in the art may make various modifications and substitutions to the present invention without departing from the purpose and spirit of the present invention.

[0039] Unless otherwise specified, the experimental methods used in the following examples are conventional methods.

[0040] Unless otherwise specified, the materials and reagents used in the following examples can be obtained from commercial sources.

[0041] Example 1

[0042] The preparation method of the semaglutide-counterion complex used in this example is as follows: weigh semaglutide, and prepare a hydrochloric acid solution of semaglutide using 0.01 mol / L HCl solution as a solvent; weigh the counterion, add an appropriate amount of ultrapure water to prepare a counterion solution; add the counterion solution to the semaglutide solution according to the molar ratio, and the mixture is shaken at 150 rpm at room temperature for 2 hours until the solution becomes white turbid; centrifuge the suspension at 11220 rpm for 5 minutes, discard the supernatant, wash the precipitate with ultrapure water 2-3 times, collect the precipitate, and freeze-dry. The final white powder is the semaglutide-counterion complex, which is collected in a vial and stored in a sealed dark place in a refrigerator at 4°C for later use.

[0043] 1. Investigation of the recombination rate of semaglutide and different counterions

[0044] Semaglutide was dissolved in 0.01 mol / L HCl to prepare a 1 mg / mL solution. The counterions sodium docusate, sodium lauryl sulfate, sodium lauryl sulfonate, sodium oleate, and sodium deoxycholate were weighed and dissolved in water at molar ratios of 1:1, 2:1, 4:1, and 6:1 to the semaglutide solution. The counterion solution was added dropwise to the semaglutide solution at a 1:1 (v / v) ratio under gentle shaking. The mixture was shaken at 150 rpm for 2 h at room temperature. The suspension was centrifuged at 11220 rpm for 10 min. An appropriate amount of the supernatant was collected and filtered. The semaglutide concentration was determined by HPLC. The semaglutide conjugation rate was calculated based on the initial amount of semaglutide added.

[0045] like Figure 1As shown, sodium oleate and sodium deoxycholate did not result in the formation of any ion pairs. This is because the ionizable structures of sodium oleate and sodium deoxycholate are weakly acidic carboxylic acid groups, which gain protons and lose charge at low pH. The ionizable structures of sodium dodecyl sulfate, sodium dodecyl sulfonate, and docusate sodium are highly acidic sulfate and sulfonic acid groups, which retain negative charges at low pH. All of them can form complexes with semaglutide and precipitate. Among the three, the complexation rate of docusate sodium with semaglutide at various reaction molar ratios is higher than that of sodium dodecyl sulfate and sodium dodecyl sulfonate.

[0046] 2. Determination of logP of semaglutide-counterion complex

[0047] Semaglutide was dissolved in 0.01 mol / L HCl to prepare a 1 mg / mL solution. The counterions sodium docusate, sodium lauryl sulfate, and sodium lauryl sulfonate were weighed and dissolved in water at molar ratios of 2:1, 4:1, and 6:1 to the semaglutide solution. The counterion solution was added dropwise to the semaglutide solution at a 1:1 (v / v) ratio under gentle shaking. The mixture was shaken at 150 rpm for 2 hours at room temperature. The suspension was centrifuged at 11220 rpm for 10 minutes, the supernatant was decanted, the precipitate was washed twice with purified water, and then lyophilized. The lyophilized complex and semaglutide API were then added with 2 mL of n-octanol and 2 mL of purified water, respectively. The mixture was placed on a thermostatic shaker (100 rpm, 37°C) for 24 hours and then centrifuged at 8000 rpm for 15 minutes for phase separation. The n-octanol layer and the aqueous layer were collected, filtered, and the amounts of semaglutide in the n-octanol and aqueous phases were determined by HPLC. The logP values ​​of the complexes and semaglutide were calculated according to the formula.

[0048] The oil-water partition coefficient (P) of a drug is the concentration ratio when the drug reaches distribution equilibrium between the oil phase (such as n-octanol) and the water phase. It is usually expressed as a logarithmic value (Log P) and is a key parameter in drug development. The Log P value reflects the distribution of the drug between the oil and water phases. The larger the Log P value, the more lipophilic the drug is and the more conducive it is to loading into SEDDS. The HIP complex has sufficient lipophilicity (Log P>2) and can be stably added to SEDDS. Figure 2As shown, the Log P of semaglutide is -0.95, and complexation with each counterion significantly enhances its lipid solubility. DOC enhances the lipid solubility of semaglutide more effectively than the other two counterions at all molar ratios, with the SET-DOC complex achieving the highest logP value of 2.16 at a 4:1 reaction molar ratio. DOC is the most commonly used surfactant in HIPs and can form HIPs with a variety of drugs, including peptides and proteins. Therefore, subsequent experiments used the SET-DOC complex formed by reacting docusate sodium with semaglutide at a 4:1 molar ratio.

[0049] 3. Differential Scanning Calorimetry

[0050] like Figure 3 As shown, semaglutide (SET) exhibits a large endothermic peak at 88°C and a short endothermic peak at 179.4°C. Docusate sodium (DOC) exhibits a small endothermic peak at 91.5°C, and a strong, broad endothermic peak appears above 250°C. The thermal curve of the SET / DOC mixture largely retains the characteristic endothermic peaks corresponding to both SET and DOC, with slight differences in their intensity and location. The spectrum of the SET-DOC complex reveals only shallow endothermic peaks at 275.25°C and 282.29°C, and a small exothermic peak at 279.95°C. The original endothermic peaks of SET at 88°C and 179.4°C, and the original endothermic peak of DOC at 91.5°C, disappear. These results indicate that DOC and SET are not simply physically mixed, but rather interact to form a complex.

[0051] Example 2

[0052] 1. Determination of the equilibrium solubility of semaglutide-docusate sodium ion pairs in oily solvents

[0053] 1.0 g of various oily solvents were weighed into vials (n = 3). The types of oily solvents are listed in Table 1. The SET-DOC complex was added to the oily solvents until excess; the mixture was vortexed and placed in a 37°C constant temperature shaker at 100 rpm for 48 hours. Samples were collected and centrifuged at 8000 rpm for 15 minutes. The supernatant was diluted with methanol to an appropriate multiple. The equilibrium solubility of the HIP complex in various oily solvents was determined by HPLC.

[0054] Table 1. Types of oily solvents

[0055]

[0056] The carbon chain length, unsaturation and presence of polar functional groups in the oil phase will affect its interaction with the HIP complex molecules, thus affecting the solubility. Figure 4-6 As shown, the saturated solubility of the SET-DOC complex in the medium-chain glyceride Capryol 90 was the highest, at (1.87±0.10) mg / g. Plurol Oleique CC 497 ranked second, followed by Peceol. Among the screened emulsifiers, Labrasol had the highest solubility for the SET-DOC complex, with an equilibrium solubility of (2.18±0.05) mg / g. The other emulsifiers exhibited lower solubility. Among the screened cosolvents, Transcutol HP exhibited the strongest solubility for the SET-DOC complex, at (11.70±0.32) mg / g.

[0057] 2. Compatibility study of mixed oil phase and Labrasol

[0058] 2.0g each of Capryo 90 and WL 1349 were mixed to form the mixed oil phase. Capryo 90, a 1:1 (w / w) mixture of Capryo 90 and WL 1349, and Labrasol were then mixed in ratios of 1:9, 2:8, 3:7, and 4:6, respectively. 200 volumes of deionized water were then added to the mixture and gently stirred at 100 rpm using a magnetic stirrer at 37°C. The self-emulsification time was measured, and the self-emulsification performance was visually observed. The results were then graded into five grades: A, B, C, D, and E.

[0059] The specific classification standards for self-emulsification effects are as follows:

[0060] A: Microemulsion forms very quickly (less than 1 minute), and the liquid is clear or has a blue opalescence.

[0061] B: Microemulsion forms quickly (within 1 minute), and the liquid is bluish-white;

[0062] C: White emulsion formed quickly (within 2 minutes) and was bright white in color;

[0063] D: The emulsification time is long (more than 2 minutes), and the liquid surface has a slight oil droplet and is dark grayish white;

[0064] E: Emulsification is difficult, and large unemulsified oil droplets continue to exist on the liquid surface.

[0065] As shown in Table 2, the addition of WL 1349 can significantly improve the compatibility of the Capryo 90:Labrasol system. Therefore, Capryo 90:WL 1349 was selected as the oil phase, Labrasol as the surfactant, and Transcutol HP as the cosolvent for subsequent formulation screening.

[0066] Table 2. Compatibility of mixed oil phase with Labrasol

[0067]

[0068] 3. Drawing of pseudo-ternary phase diagram

[0069] Based on compatibility studies, Capryo 90:WL 1345 was selected as the oil phase, Labrasol as the surfactant, and Transcutol HP as the cosolvent. Capryo 90 and WL 1345 were first mixed uniformly in mass ratios of 2:1, 1:1, 1:2, and 1:3 to form the mixed oil phase. The emulsifier, Labrasol, and the cosolvent, Transcutol HP, were then mixed uniformly in mass ratios of 1:9, 2:8, 3:7, 4:6, 5:5, 6:4, 7:3, 8:2, and 9:1 to form the mixed surfactant. The mixed oil phases in each ratio were then compounded with the mixed surfactants in each ratio in mass ratios of 1:9, 2:8, 3:7, 4:6, 5:5, and 6:4, respectively, with the total mass of each formulation being fixed at 1.0 g. After vortex mixing for 5 minutes, an appropriate amount of each formulation was added to 200 times deionized water and stirred gently at 37°C and 100 rpm using a magnetic stirrer. The emulsified solution was observed and the formulation that appeared clear, (semi-)transparent and had a light blue opalescence was placed in the pseudo-ternary phase diagram to draw the pseudo-ternary phase diagram.

[0070] like Figure 7 As shown, a 1:2 Capryo 90:WL 1345 ratio achieves the largest self-emulsification region. A 1:3 Capryo 90:WL 1345 ratio shows a slightly smaller self-emulsification region than a 1:2 ratio. This indicates that a 1:2 Capryo 90:WL 1345 ratio achieves the best self-emulsification effect. Considering the effect of adding WL1349 on solubility, a 1:2 Capryo 90:WL 1345 ratio was selected as the oil phase for the next step of screening.

[0071] 4. Screening of oil phase ratio

[0072] Based on the pseudo-ternary phase diagram screening results, a Capryo 90:WL 1349 (1:2):Labrasol:Transcutol HP system was selected as the basic formulation for single-factor screening. The mass ratio (Km) of the emulsifier Labrasol to the cosolvent Transcutol HP was determined to be 2.5:1, and the oil phase ratio was screened within the range of 10% to 40%. A blank microemulsion was prepared and diluted 200-fold in deionized water. The particle size and PDI value after emulsification were measured to determine the optimal oil phase ratio.

[0073] like Figure 8 As shown, when the oil phase ratio in the formulation is less than 30%, the particle size and PDI gradually decrease as the oil phase ratio increases. When the oil phase ratio is 30%, the particle size and PDI are minimized, reaching (82.86±0.31) nm and 0.133±0.009, respectively, indicating the formation of a relatively uniform nanoemulsion. When the oil phase ratio is greater than 30%, the particle size and PDI gradually increase with increasing oil phase ratio, and some larger oil droplets remain unemulsified. Since microemulsion particle size significantly influences absorption, smaller particle size facilitates absorption. However, the oil phase ratio affects drug loading, so the final oil phase Capryo 90 / WL 1349 (1:2) ratio was determined to be 30%.

[0074] 5. Screening of emulsifier and co-solvent ratio

[0075] Blank SME systems were prepared using a 30% fixed oil phase Capryo 90:WL 1349 (1:2) ratio. Emulsifier:cosolvent mass ratios (Km) of 1:2, 1:1, 1.5:1, 2:1, 2.5:1, 3:1, and 4:1 were used. These blank SME systems were then diluted 200-fold with deionized water, and the particle size and PDI values ​​after emulsification were measured. The optimal mass ratio of emulsifier:cosolvent was determined based on these experimental results.

[0076] like Figure 9 As shown, when the emulsifier-to-co-emulsifier ratio (Km) varies from 1:2 to 4:1, the particle size and PDI of the self-emulsified formulation first decrease and then increase as Km increases. When Km = 2.5:1, i.e., when the emulsifier ratio is 50% and the co-solvent ratio is 20% in the self-emulsified formulation, the particle size and PDI of the formulation are minimized, reaching (87.06±0.53) nm and 0.150±0.005, respectively. This indicates that at this Km value, the system achieves the best self-emulsification performance. Therefore, the optimal blank SEDDS formulation is determined to be Capryo90 / WL 1349 (1:2): Labrasol: Transcutol HP = 3:5:2.

[0077] 6. Determination of drug loading of semaglutide-docusate sodium ions in self-microemulsion

[0078] The prescribed amount of blank excipients (Capryo 90 / WL 1349 (1:2): Labrasol: Transcutol HP = 3:5:2) was mixed thoroughly, and an excess of semaglutide-docusate sodium ion-pair complex was added. The mixture was vortexed for 5 minutes and shaken at 100 rpm in a 37°C constant-temperature shaker for 48 hours. The mixture was then centrifuged at 8000 rpm for 15 minutes and incubated at 37°C for 1 hour to obtain SET-DOC SEDDS. 0.2 g of the supernatant was diluted to 4 mL with methanol. The saturated solubility of SET-DOC in the self-nanoemulsion was determined and calculated by HPLC. The saturated drug loading was 2.64 ± 0.85 mg / g.

[0079] 7. Formula of semaglutide-docusate sodium ion pair self-microemulsion (mass percentage)

[0080]

[0081] The above-prescribed amount of excipients was taken and vortexed for 5 minutes to mix evenly, the prescribed amount of semaglutide-docusate sodium complex was added, vortexed for 3 minutes to mix evenly, 100 times the amount of 37°C ultrapure water was added to the mixture, and magnetic stirring was carried out at a constant speed of 200 r / min to obtain a semaglutide-docusate sodium ion pair self-microemulsion.

[0082] The particle size, distribution and Zeta potential of the self-microemulsification were investigated using a Malvern Nano ZS particle size analyzer.

[0083] Table 3. Self-emulsification efficiency, particle size, PDI and potential

[0084] Self-emulsifying time(s) Particle size (nm) PDI Zeta potential (mV) 38±0.78 78.88±0.03 0.156±0.045 -9.64±0.13

[0085] 8. In vitro release of semaglutide-docusate sodium ions from self-microemulsions

[0086] 1 mg of semaglutide API and semaglutide-docusate sodium self-microemulsion (equivalent to 1 mg of semaglutide) were placed in 50 mL of artificial intestinal fluid (pH 6.8) in a release medium in triplicate, kept in the dark, at 37 ° C and at a speed of 100 rpm. At different time points of 1, 2, 4, 6, 8, 12, and 24 h, 0.5 mL of sample was taken, and an equal amount of blank release medium was added at the same temperature. The sample was placed in an ultrafiltration tube and centrifuged at 8000 rpm for 5 min, and the filtrate was taken. The concentration of semaglutide was determined by HPLC, the cumulative release of semaglutide at each time point was calculated, and the cumulative release curve was plotted.

[0087] like Figure 10As shown in the figure, the release rate of semaglutide API was relatively fast, with approximately 99.56% released in the first 2 hours in the simulated intestinal fluid. The release rate of SET-DOC SEDDS was relatively slow in the simulated intestinal fluid, with the cumulative release reaching nearly 80% in 12 hours.

[0088] Example 3

[0089] Study on the cytotoxicity of semaglutide-docusate sodium ions on self-microemulsion

[0090] Caco-2 cells in the logarithmic growth phase were taken and 1×10 4 Cells were seeded in a 96-well plate, with control wells (cells only, no drug) and zero-wells (no cells, no drug, or medium only). After 24 hours of incubation at 37°C and 5% CO₂, the culture medium was removed and 100 μL of culture medium containing drug (free semaglutide group and SET-DOC SEDDS group) was added, resulting in final drug concentrations of 5, 10, 20, 50, and 100 μg / mL, respectively. After 12 hours of incubation in an incubator, the old culture medium was discarded, the plates were washed with PBS, and 100 μL of 0.5 mg / mL MTT solution was added to each well and incubated for 4 hours. After incubation, the MTT supernatant was discarded, and 150 μL of DMSO was added and shaken for 10 minutes to fully dissolve the crystal violet. The absorbance of each well was measured at 570 nm using a microplate reader, and cell viability was calculated according to the formula.

[0091] like Figure 11 As shown, after SET and SET-DOC SEDDS were co-incubated with Caco-2 cells, the cell activity was not affected and the survival rate was greater than 90%, indicating that the self-microemulsification preparation had good cell biosafety and compatibility.

[0092] Example 4

[0093] Evaluation of the efficacy of semaglutide-docusate sodium ion self-microemulsion

[0094] 1. Modeling

[0095] Eight-week-old male Sprague-Dawley rats, weighing 180-230g, were fed a high-fat diet with a 45% fat content for one month to induce insulin resistance. One month later, the rats were fasted overnight but not water for 16 hours and then received a single intraperitoneal injection of 2% STZ at 30mg / kg. Following the injection, the rats were given ample water and fasted for another 2 hours before resuming a high-fat diet and water.

[0096] 2. Medication

[0097] Nine SD rats with successful modeling were randomly divided into three groups (n=3): NaCl group (blank control saline group); Solution group (positive control semaglutide solution group); SEDDS group (semaglutide-docusate sodium ion pair self-microemulsion).

[0098] The treatment group received oral gavage of a self-microemulsion (SET 700 μg / kg); the control group received a SC injection (SET 70 μg / kg in normal saline); and the blank control group received oral normal saline (1 mL / kg). Dosing was performed once daily, with a 12-hour pre-dose fast (overnight fasting) and the blood glucose level was measured daily.

[0099] like Figure 12 As shown, blood glucose levels in the oral saline group remained stable with no significant fluctuations. After 15 consecutive days of treatment, blood glucose levels at fixed times of day in both the subcutaneous semaglutide and SET-DOC SEDDS groups showed an overall downward trend. The SET-DOC SEDDS group showed a significant reduction in blood glucose compared to the oral saline group.

Claims

1. A semaglutide hydrophobic ion pair complex, characterized in that: It is generated by the reaction of semaglutide and counterion, wherein the counterion is docusate sodium, sodium lauryl sulfate or sodium lauryl sulfonate; the molar ratio of semaglutide to counterion is 1:1-6:

1.

2. The semaglutide hydrophobic ion pair complex according to claim 1, characterized in that The counterion is docusate sodium.

3. A semaglutide self-emulsifying composition, characterized in that: The raw materials include an active ingredient and an excipient mixture; the active ingredient is the semaglutide hydrophobic ion pair complex according to any one of claims 1 or 2, and the excipient mixture comprises an oil phase, an emulsifier and a cosolvent; The amount of the semaglutide hydrophobic ion pair complex is 0.01-0.27% of the mass of the excipient mixture.

4. The semaglutide self-emulsifying composition according to claim 3, characterized in that The oil phase is a mixture of propylene glycol monocaprylate and medium chain triglycerides.

5. The semaglutide self-emulsifying composition according to claim 4, characterized in that The oil phase accounts for 10%-40% of the total mass of the auxiliary material mixture.

6. The semaglutide self-emulsifying composition according to claim 4, characterized in that The mass ratio of the propylene glycol monocaprylate to the medium chain triglyceride is 2:1-1:

3.

7. The semaglutide self-emulsifying composition according to claim 3, characterized in that The emulsifier is caprylic acid capric acid macrogol glyceride, and the co-solvent is diethylene glycol monoethyl ether.

8. The semaglutide self-emulsifying composition according to claim 7, characterized in that The mass ratio of the emulsifier to the co-solvent is 1:2-4:

1.

9. A semaglutide self-microemulsion, characterized in that: The method comprises the semaglutide self-emulsifying composition according to any one of claims 3 to 8 and water.

10. Use of the semaglutide hydrophobic ion pair complex according to any one of claims 1 or 2, the semaglutide self-emulsifying composition according to any one of claims 3 or 8, or the semaglutide self-microemulsification according to claim 9 in the preparation of a medicament for preventing and treating diabetes, obesity, non-alcoholic fatty liver disease, or neurodegenerative diseases.

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