Pharmaceutical composition of stable receptor agonist, preparation method and application thereof

Abstract

The present invention relates to a pharmaceutical composition of a stable receptor agonist, a preparation method thereof and its application. Specifically, the present invention discloses a pharmaceutical composition having a dual receptor agonist as an active ingredient. The pharmaceutical composition includes a dual receptor agonist buffer, an osmotic pressure regulator and a pH regulator. The pharmaceutical composition of the present invention has good drug stability and safety, and the preparation method is simple and suitable for industrial production.

Description

[Technical Field]

[0001] The present invention relates to the field of pharmaceutical preparations. Specifically, the present invention relates to stable pharmaceutical compositions of GLP-1 / GIP receptor agonists, methods for their preparation, and uses. [Background technology]

[0002] Over the past 50 years, the prevalence of diabetes has continued to increase worldwide, with the number of affected individuals expected to reach 700 million by 2050. The prevalence of diabetes in China is 11.2%, affecting approximately 130 million people. China currently has the highest number of diabetes patients in the world. This has become a major public health problem, resulting in significant physical, mental, and economic burdens for patients, their families, and society. Type 2 diabetes mellitus (T2DM), which accounts for over 90% of the diabetic population, is primarily pathophysiologically characterized by a reduced ability of insulin to regulate glucose metabolism (i.e., insulin resistance) and a concomitant decrease in insulin secretion caused by pancreatic beta-cell dysfunction.

[0003] T2DM medications are classified into three major categories: oral hypoglycemic agents, GLP-1 receptor agonists (GLP-1 RAs), and insulin / insulin analogs. Oral hypoglycemic agents mainly include insulin secretagogues (sulfonylureas and non-sulfonylureas) and insulin sensitizers (metformin and thiazolidinediones). Metformin is the first-line treatment for clinical use, boasting a potent hypoglycemic effect, a low risk of hypoglycemia, and the benefits of weight loss. When glycemic control is poor, metformin is often combined with insulin secretagogues, dipeptidyl peptidase-4 (DPP-4) inhibitors, GLP-1 RAs, or other drugs for dual or triple therapy. Obese T2DM patients may undergo metabolic surgery to alleviate or even reverse their diabetic condition while achieving significant weight loss. After metabolic surgery, gut hormones are significantly altered, and the incretin secretion effect is enhanced, achieving metabolic benefits such as weight loss and blood glucose reduction. The incretin secretory effect is one of the main causes of the postprandial increase in insulin secretion in healthy populations, where two gut hormones that play a key role are GLP-1 and GIP, respectively.

[0004] After meals, T2DM patients experience reduced secretion of GLP-1. GLP-1 binds to receptors and then exerts insulin secretion-stimulating effects similar to those in healthy individuals. GLP-1 in the human body has a very short half-life due to its rapid degradation by DPP-4. To develop drugs based on this mechanism, endogenous human GLP-1 is restructured to function with a longer half-life in the human body. In recent decades, the development of GLP-1 RAs has progressed rapidly. In addition to their clear blood glucose-lowering effects, they also have a clear weight-loss benefit. Compared with placebo, they can also significantly reduce the risk of major cardiovascular events and all-cause mortality. In addition to treating diabetes, they are also being developed for the treatment of overweight or obese non-diabetic patients. Among GLP-1 RAs, dulaglutide and semaglutide have significant effects. Dulaglutide was marketed in China in 2019 and has significantly better blood glucose-lowering and weight-loss effects than liraglutide. Semaglutide has superior blood glucose lowering and weight loss effects to dulaglutide and has a similar safety profile to dulaglutide.

[0005] Long-term treatment with high concentrations of GLP-1 RAs can maintain favorable blood glucose lowering and weight loss effects, but severe gastrointestinal side effects, such as nausea and vomiting, limit the use of maximum doses of GLP-1 RAs, thereby preventing their maximum potential efficacy from being achieved. Therefore, a large gap remains between the efficacy of GLP-1 RAs and that of gastrointestinal metabolic surgery for T2DM. Currently, multitarget drug development strategies based on the mechanism of action of GLP-1 RAs have been proposed, and another GIP with incretin activity is a key research target. Although the blood glucose lowering effect of GIP is significantly impaired by elevated blood glucose concentrations in T2DM patients, its insulin secretion-stimulating effect can be rapidly restored when blood glucose concentrations return to normal levels. Furthermore, GIP regulates abnormal fat metabolism, reduces abnormal fat accumulation in tissues, improves insulin sensitivity, and acts synergistically with GLP-1 in the central nervous system to reduce food intake. GIP can also significantly reduce gastrointestinal reactions such as nausea and vomiting caused by the antitumor drug cisplatin, potentially further enhancing the efficacy of GLP-1 RAs by increasing their tolerability.

[0006] The active ingredient of the present invention is a new generation GIP / GLP-1 dual receptor agonist developed by the inventors, which has very strong agonist activity on both the GIP receptor and the GLP-1 receptor. Due to its dual receptor agonist activity, the dual receptor agonist is intended for the treatment of type 2 diabetes mellitus (T2DM). Pharmacodynamic experiments in animals have shown that after a single administration, the present invention can significantly reduce random blood glucose, stimulate insulin secretion, reduce food intake, and have a certain weight loss effect in db / db diabetic mice. The pharmaceutical composition of the present invention has a superior therapeutic effect to semaglutide at an equivalent dose. After long-term continuous administration once every three days, the pharmaceutical composition of the present invention can dose-dependently reduce blood glucose, body weight, and liver weight in db / db diabetic mice. In the continuous administration test, the pharmaceutical composition of the present invention can dose-dependently reduce the body weight of DIO mice, reduce food intake and liver weight, and also significantly reduce the plasma triglyceride, low-density lipoprotein cholesterol and total cholesterol levels of DIO (diet-induced obesity) mice, and improve the blood lipid metabolism disorder in DIO mice.The pharmaceutical composition of the present invention has better blood lipid lowering and weight reducing effects than the same dose of semaglutide.Therefore, it can be seen that the active ingredient of the present invention has good blood glucose lowering effect, can play a role in reducing blood lipids and body weight, and has good development prospects.

[0007] The active ingredient in the present invention is a polypeptide, which is a compound formed by linking multiple amino acids together via peptide bonds. The results of stress tests on bulk drugs show that significant decomposition occurs under acid, alkaline, oxidative, or high-temperature conditions. The results of impact factor tests on bulk drugs show that polymer growth occurs under high-temperature or light conditions. The characteristics of the active ingredient result in insufficient stability. Obtaining a stable, high-quality, industrially producible pharmaceutical composition of a GLP-1 / GIP receptor agonist remains a major challenge for researchers. Summary of the Invention

[0008] An object of the present invention is to provide a stable pharmaceutical composition of a GIP / GLP-1 dual receptor agonist.

[0009] The object of the present invention is achieved by the following technical solutions:

[0010] The pharmaceutical composition of the present invention comprises an active ingredient, a buffer, an osmotic pressure adjusting agent, and a pH adjusting agent, and the active ingredient of the present invention is [ka] It has the following structure.

[0011] The amino acid sequence of this structure is: L-Tyrosyl-isobutyryl-L-glutamyl-glycyl-L-threonyl-L-phenylalanyl-L-threonyl-L-seryl-L-aspartyl-L-tyrosyl-L-seryl-L-isoleucyl-L-tyrosyl-L-leucyl-L-glutamyl-L-lysyl-L-isoleucyl-L-alanyl-L-alanyl-L-glutaminyl-L-glutamyl-L-phenylalanyl-L-valyl-L-asparaginyl-L- Tryptophanyl-L-leucyl-L-leucyl-L-alanyl-glycyl-glycyl-L-prolyl-L-seryl-L-seryl-glycyl-L-alanyl-L-prolyl-L-prolyl-L-prolyl-L-seryl-{N6-[(22S)-10,19,24-trioxo-3,6,12,15-tetraoxa-9,18,23-triaza-22,42-dicarboxy-tetradodecanoyl]}-L-lysineamide.

[0012] Preferably, the pharmaceutical composition of the GIP / GLP-1 dual receptor agonist is an injection, and the active ingredient is at a concentration selected from 0.5 mg / mL to 40 mg / mL, preferably 1 mg / mL to 30 mg / mL, and more preferably 1 mg / mL, 2 mg / mL, 5 mg / mL, 10 mg / mL, 15 mg / mL, 20 mg / mL, 25 mg / mL, or 30 mg / mL.

[0013] Preferably, the injection contains the active ingredient at 0.05% to 4.0% (W / V), preferably 0.1% to 4.0% (W / V), more preferably 0.1% to 3.0% (W / V).

[0014] In the present invention, the unit of W / V is g / mL.

[0015] Preferably, the buffer is selected from phosphate buffer, acetate buffer, citrate buffer, carbonate buffer, tartrate buffer, Tris buffer, and histidine salts, preferably citrate buffer or phosphate buffer, more preferably disodium hydrogen phosphate.

[0016] Preferably, the injection contains 0.05% to 3.0% (W / V), preferably 0.05% to 2.0% (W / V), more preferably 0.05% to 1.0% (W / V) of a buffering agent.

[0017] Preferably, the osmolality modifier is selected from one or more of mannitol, lactose, sucrose, propylene glycol, and glycerol, preferably propylene glycol or mannitol.

[0018] Preferably, the injection contains an osmotic agent in an amount of 0.05% to 5.0% (W / V), preferably 1.0% to 3.0% (W / V), and more preferably 1.0% to 2.0% (W / V).

[0019] Preferably, the pH adjuster is selected from one or more of hydrochloric acid or sodium hydroxide.

[0020] Preferably, the injection contains 0.1% to 3.0% (W / V) of the active ingredient, 0.05% to 3.0% (W / V) of a buffer, 0.5% to 5.0% (W / V) of an osmotic pressure adjuster, and an appropriate amount of a pH adjuster.

[0021] Preferably, the injection contains 0.1% to 3.0% (W / V) of the active ingredient, 0.05% to 2.0% (W / V) of a buffer, 1.0% to 3.0% (W / V) of an osmotic pressure adjuster, and an appropriate amount of a pH adjuster.

[0022] Preferably, the injection contains 0.1% to 3.0% (W / V) of the active ingredient, 0.05% to 1.0% (W / V) of a buffer, 1.0% to 2.0% (W / V) of an osmotic pressure adjuster, and an appropriate amount of a pH adjuster.

[0023] Preferably, the injection contains 0.1% to 3.0% (W / V) of the active ingredient, 0.05% to 0.1% (W / V) of a buffering agent, 1.0% to 2.0% (W / V) of an osmotic pressure adjusting agent, and an appropriate amount of a pH adjusting agent.

[0024] Preferably, the injection contains 0.1% to 3.0% (w / v) of the active ingredient, 0.05% to 1.0% (w / v) of disodium hydrogen phosphate and / or sodium hydroxide, 1.0% to 2.0% (w / v) of propylene glycol, and an appropriate amount of a pH adjuster.

[0025] Optionally, the injectable solution further comprises a preservative selected from m-cresol, phenol, phenylcarbinol, phenethyl alcohol, parahydroxybenzoate, hydroxybenzoate, benzyl alcohol, chlorobutanol, phenoxyethanol, methylparaben, and the like, preferably m-cresol, phenol, or trichloro-tert-butanol.

[0026] Optionally, the injection contains 0.01% to 3% (W / V) of a preservative, preferably 0.05% to 1.5% (W / V), more preferably 0.1% to 1% (W / V), and even more preferably 0.1% to 0.5% (W / V).

[0027] Preferably, the injection contains 0.1% to 3.0% (W / V) of the active ingredient, 0.05% to 1.0% (W / V) of disodium hydrogen phosphate and / or sodium hydroxide, 1.0% to 2.0% (W / V) of propylene glycol, 0.05% to 1.5% (W / V) of a preservative, and an appropriate amount of a pH adjuster.

[0028] Preferably, the injection contains 0.1% to 3.0% (W / V) of the active ingredient, 0.05% to 1.0% (W / V) of disodium hydrogen phosphate and / or sodium hydroxide, 1.0% to 2.0% (W / V) of propylene glycol, 0.1% to 1% (W / V) of a preservative, and an appropriate amount of a pH adjuster.

[0029] Preferably, the injection contains 0.1% to 3.0% (W / V) of the active ingredient, 0.05% to 1.0% (W / V) of disodium hydrogen phosphate and / or sodium hydroxide, 1.0% to 2.0% (W / V) of propylene glycol, 0.1% to 0.5% (W / V) of a preservative, and an appropriate amount of a pH adjuster.

[0030] Preferably, the weight ratio of the active ingredient to the buffering agent is 1:0.01-10, preferably 1:0.02-1, more preferably 1:0.02-0.5.

[0031] Preferably, the weight ratio of the active ingredient to the osmotic pressure adjusting agent is 1:0.1-20, preferably 1:0.4-10.

[0032] The ratio of the total amount of disodium hydrogen phosphate to the total amount of sodium hydroxide in the buffer solution is preferably 1:0 to 3.0, preferably 1:0.10 to 2.5, more preferably 1:0.13 to 1.5, and even more preferably 1:0.13 to 1.2.

[0033] The weight ratio of the buffer solution to the osmotic pressure adjusting agent is preferably 1:5-80, more preferably 1:5-60, more preferably 1:15-40, and even more preferably 1:15-25.

[0034] Preferably, the pharmaceutical composition has a pH range of 6.5 to 9.0, preferably 7.0 to 8.5, more preferably 7.0 to 8.0.

[0035] Preferably, the administration volume of the pharmaceutical composition is 0.5 mL to 1 mL.

[0036] Another object of the present invention is to provide a method for preparing a pharmaceutical composition, in which a buffer and an osmolality adjuster are dissolved in water for injection, an active ingredient is dissolved in the drug solution by stirring, a pH adjuster is added, the mixture is adjusted to a specified volume, filtered, and sub-packaged.

[0037] Preferably, the water for injection is cooled to 25°C or below.

[0038] Preferably, the resulting pharmaceutical composition is sterile filtered, subpackaged, and encapsulated.

[0039] Preferably, the filling process is completed under nitrogen protection.

[0040] Preferably, the pharmaceutical composition is sub-packaged in a pre-filled syringe.

[0041] Preferably, the pharmaceutical composition is sub-packaged in a cartridge bottle.

[0042] Preferably, the pharmaceutical composition is subpackaged in a vial.

[0043] Another object of the present invention is to provide a use of a pharmaceutical composition for the manufacture of a medicament for treating non-insulin dependent diabetes mellitus, insulin dependent diabetes mellitus, obesity, insulin resistance, or blood lipid metabolism disorders, preferably wherein the non-insulin dependent diabetes mellitus is type II diabetes mellitus.

[0044] The pharmaceutical compositions of the present invention can be used simultaneously, separately, or sequentially in combination with one or more agents selected from metformin, thiazolidinediones, sulfonylureas, dipeptidyl peptidase inhibitors, and sodium glucose cotransporters.

[0045] Unlike other chemical products, even a small amount of impurities in drugs can pose risks to the safety, efficacy, quality control, etc. of clinical administration. In the present invention, a pharmaceutical composition with good stability can be obtained by controlling the components and component proportions in the formulation without adding other auxiliary substances, wherein the content of related substances is effectively controlled and the content of specific impurities is significantly reduced, thereby significantly reducing the toxicity of the drug, and the content of specific impurities increases slowly after long-term storage of the drug, thereby improving the safety of the drug. Furthermore, filling under nitrogen protection improves the stability of the preparation. DETAILED DESCRIPTION OF THE INVENTION

[0046] It should be understood that those skilled in the art can make various modifications and improvements to the present invention based on the contents disclosed herein without departing from the spirit and scope of the present invention. All various modifications and improvements should be encompassed within the scope of protection defined by the claims of this application. Furthermore, it should be understood that the examples provided herein are for the sole purpose of illustrating the present invention and should not be construed as limiting the present invention.

[0047] Example I. [Table 1] Water for injection was cooled to below 25°C and added for later use. Disodium hydrogen phosphate (0.071 g) was weighed out according to the formula and dissolved in 90 mL of water for injection, followed by stirring to dissolve. Next, 0.1 mL of 100 g / L sodium hydroxide solution was added and thoroughly stirred (0.2 g of the active ingredient was not added). Propylene glycol (1.5 g) was weighed out according to the formula and thoroughly stirred. The active ingredient of the present invention was weighed out according to the formula and slowly stirred to dissolve, and the solution was adjusted to pH 7.5-8.0 with sodium hydroxide or hydrochloric acid solution. The solution was adjusted to 100 mL with water for injection. Finally, the solution was filtered through a 0.22 μm microporous filter membrane. The filtered solution was subpackaged into prefilled syringe bottles, the bottles were filled with nitrogen, and stoppers were inserted into the bottles. Samples were tested for stability at 25°C ± 2°C / 60% RH ± 5% RH and 40°C, respectively. Test parameters included appearance and related substances, and the results are shown in the table below. [Table 2]

[0048] Example II [Table 3] The water for injection was cooled to below 25°C and added for later use. Disodium hydrogen phosphate (0.071g) was weighed according to the prescription and dissolved in 90mL of water for injection, followed by stirring to dissolve. Then, 0.1mL of 100g / L sodium hydroxide solution was added and thoroughly stirred. Propylene glycol (1.5g) was weighed according to the prescription and thoroughly stirred. The active ingredient of the present invention (2.0g) was weighed according to the prescription and slowly stirred to dissolve, and the solution was adjusted to pH 6.5, 7.0, 7.5, 7.8, 8.0, and 8.5 with sodium hydroxide or hydrochloric acid solution, respectively. Each of the above solutions was adjusted to 100mL with water for injection. Finally, the solution was filtered through a 0.22μm microporous filter membrane. The filtered solution was sub-packaged into a pre-filled syringe bottle, the bottle was filled with nitrogen, and the stopper was pressed into the bottle. The samples were tested for stability under the conditions of 25°C ± 2°C / 60% RH ± 5% RH. Test parameters included appearance and related substances, and the results are shown in the table below. [Table 4]

[0049] Example III. [Table 5] Water for injection was cooled to below 25°C and added for later use. Disodium hydrogen phosphate (0.071 g) was weighed out according to the recipe and dissolved in 90 mL of water for injection, followed by stirring to dissolve. 100 g / L sodium hydroxide solution was then added according to the recipe and thoroughly stirred. Propylene glycol (1.5 g) was weighed out according to the recipe and thoroughly stirred. The pH of the solution was determined, and the active ingredient of the present invention (2.0 g) was weighed out according to the recipe and slowly added. The mixture was stirred for 45 minutes, and the dissolution of the active ingredient was then observed. The pH of the solution was determined, and the samples were tested for stability under conditions of 25°C ± 2°C / 60% RH ± 5% RH, 5°C ± 3°C, 40°C, and lighting. The impurities in the samples were analyzed to examine the effects of various buffer concentrations on the impurities, thereby providing a research concept for selecting pharmaceutical compositions. The test results are shown in the table below. [Table 6-1] [Table 6-2]

[0050] Example IV. [Table 7] Water for injection was cooled to below 25°C and added for later use. Disodium hydrogen phosphate was weighed according to the formula and dissolved in 90 mL of water for injection, followed by stirring to dissolve. 0.1 mL of 100 g / L sodium hydroxide solution was then added and thoroughly stirred. Propylene glycol (1.5 g) was weighed according to the formula and thoroughly stirred. The active ingredient of the present invention (2.0 g) was weighed according to the formula and slowly stirred to dissolve. The solution was then adjusted to a pH of 7.5-8.0 with sodium hydroxide or hydrochloric acid solution, respectively. Each of the above solutions was adjusted to 100 mL with water for injection. Finally, the solution was filtered through a 0.22 μm microporous filter membrane. The filtered solution was subpackaged into prefilled injection bottles, the bottles were filled with nitrogen, and stoppers were inserted into the bottles. Samples were tested for stability at 25°C ± 2°C / 60% RH ± 5% RH or 40°C. Test parameters included appearance and related substances, and the results are shown in the table below. [Table 8]

[0051] Example V [Table 9] Water for injection was cooled to below 25°C and added for later use. Disodium hydrogen phosphate (0.071 g) was weighed out according to the formula and dissolved in 90 mL of water for injection, followed by stirring to dissolve. 0.1 mL of 100 g / L sodium hydroxide solution was then added and thoroughly stirred. The osmolality adjuster was weighed out according to the formula and thoroughly stirred. The active ingredient of the present invention (2.0 g) was weighed out according to the formula and slowly stirred to dissolve. The solution was adjusted to a pH of 7.5-8.0 with sodium hydroxide or hydrochloric acid solution, respectively. The solution was adjusted to 100 mL with water for injection. Finally, the solution was filtered through a 0.22 μm microporous filter membrane. The filtered solution was subpackaged into prefilled injection bottles, the bottles were filled with nitrogen, and stoppers were inserted into the bottles. Samples were tested for stability under conditions of 25°C ± 2°C / 60% RH ± 5% RH. Test parameters included appearance and related substances, and the results are shown in the table below. [Table 10]

[0052] Example VI [Table 11] Water for injection was cooled to below 25°C and added for later use. Disodium hydrogen phosphate was weighed according to the formula and dissolved in 90 mL of water for injection, followed by stirring to dissolve. 0.1 mL of 100 g / L sodium hydroxide solution was then added and thoroughly stirred. Propylene glycol was weighed according to the formula and thoroughly stirred. 2.0 g of the active ingredient of the present invention was weighed according to the formula and slowly stirred to dissolve. The solution was adjusted to a pH of 7.5-8.0 with sodium hydroxide or hydrochloric acid solution, respectively. The solution was adjusted to 100 mL with water for injection. Finally, the solution was filtered through a 0.22 μm microporous filter membrane. The filtered solution was subpackaged into prefilled injections and stoppers were inserted into them. Samples were tested for stability under conditions of 25°C ± 2°C / 60% RH ± 5% RH, 5°C ± 3°C, 40°C, lighting, and low-temperature cycling. Test parameters included appearance and related substances, and the results are shown in the table below. [Table 12-1] [Table 12-2]

[0053] Experimental Example I: Hemolysis Test 1. Test substance: Test sample: the injection prepared in Example 1 (containing 2.0 g of the active ingredient); 1.2 Vehicle: Sodium chloride injection.

[0054] 2. Experimental Method: Various amounts (0.5-0.1 mL) of test sample and various amounts (2.0-2.4 mL) of sodium chloride injection were added to 2.5 mL of 2% rabbit red blood cell suspension in glass test tubes, respectively. Sodium chloride injection and sterile water for injection were used as negative and positive controls, respectively, for a total volume of 5.0 mL per test tube. Each test tube was incubated in an electronic thermostat (set at 37°C) for 3 hours to observe red blood cell lysis and aggregation.

[0055] As can be seen from the test tubes containing 0.5-0.1 mL of test sample and the negative control test tube, red blood cells settled to the bottom of the tube, the supernatant was clear and colorless, and the red blood cells at the bottom of the tube were completely dispersed without hemolysis or agglutination after shaking. Also, as can be seen from the positive control tube containing sterile water for injection, some red blood cells settled to the bottom of the tube, the supernatant was clear and red, and the red blood cells at the bottom of the tube were partially hemolyzed and completely dispersed without agglutination after shaking.

[0056] Under these experimental conditions, 0.1 to 0.5 mL of the injectable pharmaceutical composition of the present invention at a nominal concentration of 20 mg / mL did not cause hemolysis of rabbit red blood cells or red blood cell aggregation in the experimental system.

[0057] Experimental Example II: Subcutaneous Irritation Test 1. Test substance: Test sample: the injection prepared in Example 1 (containing 2.0 g of the active ingredient); 2. Experimental Method: New Zealand rabbits were selected for the experiment, and each of them was subcutaneously administered with the test sample to the left and right sides of the body of each given rabbit for self-control, and the local subcutaneous irritation response at the administration site caused by a total of four subcutaneous injections was observed. During this experiment, no abnormalities related to the injectable pharmaceutical composition of the present invention were found at the local administration site of all animals.

[0058] Experimental Example III: Testing the therapeutic effect in type 2 diabetic db / db mice after a single subcutaneous injection of the pharmaceutical composition of the present application 1. Test substance: Test sample: the injection prepared in Example 1 (containing 2.0 g of the active ingredient); 2. Experimental Method The effects of a single administration on blood glucose, serum insulin levels, body weight, and food intake in type 2 diabetic db / db mice were examined, while observing the effective period after a single administration. The experimental results showed that the pharmaceutical composition could significantly lower random blood glucose in db / db mice after a single subcutaneous injection at doses of 1, 3, or 10 nmol / kg, demonstrating a good dose-effect relationship and a clear random blood glucose lowering effect at a dose of 3 nmol / kg. The duration of the blood glucose lowering effect of the pharmaceutical composition after a single subcutaneous injection was dose-dependent, with the random blood glucose lowering effect lasting 1 day at a dose of 1 nmol / kg and 2 to 3 days at doses of 3 nmol / kg and 10 nmol / kg. [Table 13]

[0059] Furthermore, using a mouse insulin ELISA kit, serum insulin levels in each group of db / db mice were determined 2 hours after administration. The results showed that after a single subcutaneous injection of the pharmaceutical composition into db / db mice at 1, 3, and 10 nmol / kg, serum insulin levels in mice of each dose group were significantly increased compared with those of the vehicle group, indicating a good dose-effect relationship. Furthermore, the increase in serum insulin is also positively correlated with a decrease in blood glucose. [Table 14]

[0060] Experimental Example IV: Testing the therapeutic effect in type 2 diabetic db / db mice after chronic subcutaneous injection of the pharmaceutical composition of the present application 1. Test substance: Test sample: the injection prepared in Example 1 (containing 2.0 g of the active ingredient); 2. Experimental Method: Type 2 diabetic db / db mice were treated with 1000mg of benzodiazepine once every 3 days for 9 consecutive doses. After the experiment (day 26), the liver weights of db / db mice in the 1, 3, and 10 nmol / kg groups were 2.313 ± 0.113 g, 2.267 ± 0.108 g, and 1.993 ± 0.104 g, respectively, all of which were significantly reduced compared with those in the vehicle group (p < 0.05, p < 0.05, p < 0.001). There was no statistically significant difference in pancreas weight compared with the vehicle group (p > 0.05).

[0061] In conclusion, the pharmaceutical composition can dose-dependently reduce the body weight and liver weight of type 2 diabetic db / db mice by long-term continuous administration once every three days, and has a clear random blood glucose lowering effect at doses of 3 and 10 nmol / kg.

[0062] Experimental Example V: After a single subcutaneous or intravenous injection of the pharmaceutical composition of the present application Cynomolgus monkeys Pharmacokinetic studies in 1. Test substance: Injection prepared in Example 3; 2. Experimental Method: The doses administered in the four groups, i.e., intravenous injection group, low-dose subcutaneous injection group, medium-dose subcutaneous injection group, and high-dose subcutaneous injection group, were 0.1, 0.03, 0.1, and 0.3 mg / kg, respectively, and all were administered as a single dose.

[0063] After intravenous administration, the drug of the present invention showed an elimination half-life (t1 / 2) of 50.7 to 51.5 hours in cynomolgus monkeys, indicating slow elimination. It also showed a clearance rate (CL) of 0.89 to 0.93 mL / h / kg, indicating low clearance. It also showed an apparent volume of distribution (Vd) of 66.0 to 68.1 mL / kg, indicating that the drug was distributed mainly in plasma. After a single subcutaneous administration, the drug of the present invention showed a time to maximum plasma concentration (Tmax) of 13.3 to 18.0 hours in cynomolgus monkeys. It also showed a plasma elimination half-life (T1 / 2) of 48.9 to 65.4 hours, indicating slow elimination. There was no statistical difference in the exposure (AUClast) of the drug of the present invention between animals of different genders (P>0.05). Within the dose range of 0.02 to 0.2 mg / kg, the exposure of the pharmaceutical composition in the plasma of animals (AUC last ) increased with increasing dose, and the rate of increase in exposure was similar to the rate of increase in dose.

Claims

1. A stable pharmaceutical composition of a GIP / GLP-1 dual receptor agonist, comprising an active ingredient, a buffer, an osmolality adjuster, and a pH adjuster, wherein the active ingredient is 【Chemistry 1】 The pharmaceutical composition has the structure:

2. 2. The pharmaceutical composition according to claim 1, wherein the pharmaceutical composition is an injection and the active ingredient is at a concentration selected from 0.5 mg / mL to 40 mg / mL, preferably 1 mg / mL to 30 mg / mL.

3. 2. The pharmaceutical composition according to claim 1, wherein the pharmaceutical composition contains the active ingredient in an amount of 0.05% to 4.0% (W / V), preferably 0.1% to 3.0% (W / V).

4. 2. The pharmaceutical composition of claim 1, wherein the buffer is selected from phosphate buffer, acetate buffer, citrate buffer, carbonate buffer, tartrate buffer, Tris buffer, and histidine salt, preferably citrate buffer or phosphate buffer, more preferably disodium hydrogen phosphate.

5. 2. The pharmaceutical composition according to claim 1, wherein the pharmaceutical composition comprises the buffering agent in an amount of 0.05% to 3.0% (W / V), preferably 0.05% to 2.0% (W / V), more preferably 0.05% to 1.0% (W / V).

6. 2. The pharmaceutical composition of claim 1, wherein the osmolality adjuster is selected from one or more of mannitol, lactose, sucrose, propylene glycol, and glycerol, preferably propylene glycol or mannitol.

7. 2. The pharmaceutical composition according to claim 1, wherein the injectable solution contains the osmotic agent in an amount of 0.05% to 5.0% (W / V), preferably 1.0% to 3.0% (W / V), and more preferably 1.0% to 2.0% (W / V).

8. 10. The pharmaceutical composition of claim 1, wherein the pH adjusting agent is selected from one or more of hydrochloric acid or sodium hydroxide.

9. the pharmaceutical composition comprises 0.1% to 3.0% (W / V) of the active ingredient, 0.05% to 3.0% (W / V) of the buffer, 0.5% to 5.0% (W / V) of the osmotic pressure adjusting agent, and an appropriate amount of the pH adjusting agent; Preferably, the pharmaceutical composition comprises 0.1% to 3.0% (w / v) of the active ingredient, 0.05% to 2.0% (w / v) of the buffering agent, 1.0% to 3.0% (w / v) of the osmotic pressure adjusting agent, and an appropriate amount of the pH adjusting agent; More preferably, the pharmaceutical composition comprises 0.1% to 3.0% (w / v) of the active ingredient, 0.05% to 1.0% (w / v) of the buffering agent, 1.0% to 2.0% (w / v) of the osmotic agent, and an appropriate amount of the pH adjusting agent; More preferably, the pharmaceutical composition according to claim 1 comprises 0.1% to 3.0% (w / v) of the active ingredient, 0.05% to 1.0% (w / v) of disodium hydrogen phosphate and / or sodium hydroxide, 1.0% to 2.0% (w / v) of propylene glycol, and an appropriate amount of the pH adjuster.

10. 2. The pharmaceutical composition according to claim 1, wherein the weight ratio of the active ingredient to the buffering agent is 1:0.01-10, preferably 1:0.02-1, more preferably 1:0.02-0.

5.

11. 2. The pharmaceutical composition according to claim 1, wherein the weight ratio of the active ingredient to the osmotic agent is 1:0.1-20, preferably 1:0.4-10.

12. 10. The pharmaceutical composition according to claim 9, wherein the buffer comprises disodium hydrogen phosphate and sodium hydroxide, and the weight ratio of disodium hydrogen phosphate to sodium hydroxide is 1:0-3.0, preferably 1:0.10-2.5, more preferably 1:0.13-1.5, and even more preferably 1:0.13-1.

2.

13. 2. The pharmaceutical composition according to claim 1, wherein the weight ratio of the buffer to the osmotic pressure adjusting agent is 1:5-80, preferably 1:5-60, more preferably 1:15-40, and even more preferably 1:15-25.

14. 2. The pharmaceutical composition according to claim 1, wherein the pharmaceutical composition is in the pH range of 6.5 to 9.0, preferably 7.0 to 8.5, more preferably 7.0 to 8.

0.

15. 15. The pharmaceutical composition of claims 1-14, wherein the pharmaceutical composition further comprises a preservative selected from one or more of m-cresol, phenol, phenylcarbinol, phenethyl alcohol, parahydroxybenzoate, hydroxybenzoate, benzyl alcohol, chlorobutanol, phenoxyethanol, and methylparaben.

16. 16. A method for preparing the pharmaceutical composition of claims 1 to 15, wherein the buffer and the osmolality adjuster are dissolved in water for injection, the active ingredient is dissolved in the drug solution by stirring, the pH adjuster is added, the mixture is adjusted to a specified volume, filtered, and sub-packaged.

17. Use of the pharmaceutical composition according to claims 1 to 15 for the manufacture of a medicament for treating non-insulin dependent diabetes mellitus, insulin dependent diabetes mellitus, obesity, insulin resistance, or blood lipid metabolism disorders, preferably wherein the non-insulin dependent diabetes mellitus is type II diabetes mellitus.

Images