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 drug formulations. In particular, the present invention relates to a stable GLP-1 / GIP receptor agonist pharmaceutical composition, preparation method and application thereof. [Background technology]

[0002] Over the past 50 years, the prevalence of diabetes has been rising worldwide, and the number of patients is expected to reach 700 million by 2050. The prevalence of diabetes in Japan has reached 11.2%, with approximately 130 million patients. Currently, Japan is the country with the largest number of diabetes patients, and it has already become one of the major public health problems, bringing huge physiological, psychological and economic burdens to patients, families and society. Type 2 diabetes mellitus (T2DM) accounts for more than 90% of diabetes cases, and its main pathophysiological characteristics are a decrease in the ability of insulin to regulate glucose metabolism (i.e., insulin resistance) and a decrease in insulin secretion due to the dysfunction of pancreatic islet β cells.

[0003] T2DM drug treatment is mainly divided into three types: oral hypoglycemic agents, GLP-1 receptor agonists (GLP-1RAs) and insulin / insulin analogs. Oral hypoglycemic agents mainly include insulin secretagogues (sulfonylureas, non-sulfonylureas), insulin sensitizers (metformin, thiazolidinedione), etc. In clinical treatment, metformin is preferentially selected, which has a strong hypoglycemic effect, low risk of hypoglycemia, and the advantage of weight loss. In cases of poor glycemic control, two-drug or three-drug combination treatment is used, and insulin secretagogues, dipeptidyl peptidase-4 (DPP-4) inhibitors, GLP-1RAs, etc. are used in combination. For patients with obvious obesity and T2DM, metabolic surgery can be performed to obviously reduce weight and alleviate or even reverse the symptoms of diabetes. After metabolic surgery, intestinal hormones are obviously changed, and the metabolic effects such as weight loss and blood glucose reduction are achieved by enhancing the incretin secretion effect. The incretin secretion effect is one of the main reasons why insulin secretion increases after a meal in healthy people, and here, two gut hormones that play an important role are GLP-1 and GIP, respectively.

[0004] After T2DM patients eat, the secretion of GLP-1 in the body is reduced, but after binding to the receptor, the insulin secretion-promoting effect is similar to that of healthy people. GLP-1 in the human body is rapidly degraded by DPP-4, so its half-life is very short. In drug research and development for this mechanism, the structure of endogenous GLP-1 in the human body is modified to have a longer half-life in the human body and exert its effect. In the past decade, the research and development of GLP-1RAs drugs has progressed rapidly, and in addition to its remarkable blood glucose lowering effect, it also has a clear advantage in weight reduction, and can clearly reduce the risk of major cardiovascular events and all-cause mortality compared to the placebo group. In addition to being used for the treatment of diabetes, it has also been developed for the treatment of overweight and obese non-diabetic patients. Among GLP1-RAs, the effects of dulaglutide and semaglutide are remarkable, where dulaglutide was launched in China in 2019, and its blood glucose lowering and weight reduction effects are clearly superior to liraglutide. Semaglutide's blood sugar lowering and weight loss effects are superior to those of dulaglutide, and its safety profile is similar to that of dulaglutide.

[0005] Long-term high-concentration GLP-1RAs treatment can maintain good blood glucose lowering and weight reduction effects, but severe gastrointestinal side effects, such as nausea and vomiting, limit the use of the maximum dose of GLP-1RAs and prevent the maximum potential therapeutic effect from being achieved, so the therapeutic effect of GLP-1RAs on T2DM is still significantly different from that of gastrointestinal metabolic surgery. At present, a multi-target drug development policy based on the mechanism of action of GLP-1RAs has been put forward, in which GIP, which has another incretin effect, is an important research target. For GIP, in the case of high blood glucose concentration in the body of T2DM patients, its blood glucose lowering effect is severely impaired, but after the blood glucose concentration returns to normal levels, its insulin secretion promoting effect can be quickly restored. And GIP can improve insulin sensitivity by regulating abnormal fat metabolism and reducing abnormal fat accumulation between each tissue, and act on the central nervous system in cooperation with GLP-1 to reduce food intake. GIP can also significantly reduce gastrointestinal reactions such as nausea and vomiting induced by the antitumor drug cisplatin, and may further improve the therapeutic effect by increasing the tolerance of GLP-1RAs.

[0006] The active ingredient of the present invention is a new generation GIP / GLP-1 dual receptor agonist developed by the company, which has extremely strong agonist activity on the GIP receptor and also has strong agonist activity on the GLP-1 receptor, and its dual receptor agonist activity is expected to be used in the treatment of type 2 diabetes mellitus (T2DM).According to the results of animal pharmacological experiments, the present invention can significantly lower random blood glucose, promote insulin secretion, and reduce food intake in db / db diabetic mice after a single administration, and has a certain weight-lowering effect, and it is clear that the effect of the pharmaceutical composition of the present invention is superior to that of semaglutide at the same dose. By long-term continuous administration once every 3 days, it can dose-dependently reduce the blood glucose, body weight and liver weight of db / db diabetic mice; in continuous administration test, it can dose-dependently reduce the body weight, food intake and liver weight of DIO mice; it can also significantly reduce the plasma triglyceride, low-density lipoprotein cholesterol and total cholesterol levels of DIO (diet-induced obesity) mice, and improve the lipid metabolism abnormality state of DIO mice; its fat-lowering and body weight-lowering effects at the same dose are better than those of semaglutide. As can be seen, the active ingredient of the present invention has good blood glucose-lowering effect and can achieve fat-lowering and body weight-lowering effects, and has good development prospects.

[0007] The active ingredient in the present invention is a polypeptide, which is a compound formed by a plurality of amino acids linked by peptide bond.The results of the destruction experiment of the raw material show that it decomposes obviously under acid, alkali, oxidation and high temperature conditions, and the results of the influence factor experiment of the raw material show that it increases its polymer under high temperature and light irradiation conditions.Due to the special nature of the active ingredient, the stability of the active ingredient is poor, and in this case, it is still a huge challenge for researchers to obtain a stable, high-quality and industrially producible pharmaceutical composition of GLP-1 / GIP receptor agonist. Summary of the Invention

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

[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 structure of the active ingredient of the present invention is [ka] It is.

[0011] The amino acid sequence of this structure is as follows:

[0012] 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-glutamine-L-glutamyl-L-phenylalanyl-L-valyl-L-asparagine-L-thiamine Lyptophanyl-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.

[0013] Preferably, the GIP / GLP-1 dual receptor agonist pharmaceutical composition is an injection solution, and the concentration of the active ingredient is selected from 0.5 mg / mL to 40 mg / mL, preferably 1 mg / mL to 30 mg / mL, 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.

[0014] Preferably, the injection solution contains 0.05% to 4.0% (W / V) of the active ingredient, preferably 0.1% to 4.0% (W / V), more preferably 0.1% to 3.0% (W / V).

[0015] In the present invention, the unit of W / V is g / mL.

[0016] Preferably, the buffer is selected from phosphate buffer, acetate buffer, citrate buffer, carbonate buffer, tartrate buffer, Tris buffer, histidine salt, preferably citrate buffer or phosphate buffer, more preferably disodium hydrogen phosphate.

[0017] Preferably, the injection solution contains a buffer solution at 0.05% to 3.0% (W / V), preferably 0.05% to 2.0% (W / V), and more preferably 0.05% to 1.0% (W / V).

[0018] Preferably, the osmolality adjusting agent is one or more selected from mannitol, lactose, sucrose, propylene glycol, glycerin, preferably propylene glycol or mannitol.

[0019] Preferably, the injection solution contains an osmolality regulator at 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).

[0020] Preferably, the pH adjusting agent is one or more selected from hydrochloric acid, sodium hydroxide.

[0021] Preferably, the injection solution contains 0.1% to 3.0% (W / V) of the active ingredient, 0.05% to 3.0% (W / V) of a buffer solution, 0.5% to 5.0% (W / V) of an osmotic pressure adjusting agent, and an appropriate amount of a pH adjusting agent.

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

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

[0025] Preferably, the injection solution 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.

[0026] Optionally, the injection solution further comprises a preservative, the preservative being selected from m-cresol, phenol, phenylcarbinol, phenylethanol, parahydroxybenzoic acid esters, hydroxybenzoic acid esters, benzyl alcohol, chlorobutanol, phenoxyethanol, methylparaben, etc., preferably m-cresol, phenol or chlorobutanol.

[0027] Optionally, the injection solution contains 0.01% to 3% (W / V) preservative, preferably 0.05% to 1.5% (W / V) preservative, more preferably 0.1% to 1% (W / V), and even more preferably 0.1% to 0.5% (W / V) preservative.

[0028] Preferably, the injection solution 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.

[0029] Preferably, the injection solution 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.

[0030] Preferably, the injection solution 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.

[0031] Preferably, the weight ratio of the active ingredient to the buffer solution is 1:0.01-10, preferably 1:0.02-1, more preferably 1:0.02-0.5.

[0032] Preferably, the weight ratio of the active ingredient to the osmotic pressure regulator is 1:0.1-20, preferably 1:0.4-10.

[0033] Preferably, the total ratio of disodium hydrogen phosphate and sodium hydroxide in the buffer solution is 1:0-3.0, preferably 1:0.10-2.5, more preferably 1:0.13-1.5, and further preferably 1:0.13-1.2.

[0034] The weight ratio of the buffer solution to the osmotic pressure regulator is preferably 1:5-80, preferably 1:5-60, more preferably 1:15-40, and further preferably 1:15-25.

[0035] Preferably, the pH range of the pharmaceutical composition is from 6.5 to 9.0, preferably from 7.0 to 8.5, more preferably from 7.0 to 8.0.

[0036] Preferably, the dosage volume of the pharmaceutical composition is between 0.5 ml and 1 ml.

[0037] Another object of the present invention is to provide a method for preparing said pharmaceutical composition, which comprises dissolving a buffer and an osmolality adjusting agent in water for injection, dissolving an active ingredient in said liquid, stirring to dissolve, adding a pH adjusting agent, adjusting the volume to volume, filtering and dispensing.

[0038] Preferably, the water for injection is cooled to ≦25° C.

[0039] Preferably, the preparation of the pharmaceutical composition involves sterile filtration, dispensing and encapsulation.

[0040] Preferably, the filling process is completed under nitrogen gas protection.

[0041] Preferably, the pharmaceutical composition is dispensed using a pre-filled syringe.

[0042] Preferably, the pharmaceutical composition is dispensed using a card bottle.

[0043] Preferably, the pharmaceutical composition is dispensed using a penicillin bottle.

[0044] Another object of the present invention is to provide a use of the pharmaceutical composition in the manufacture of a medicament for treating non-insulin dependent diabetes mellitus, insulin dependent diabetes mellitus, obesity, insulin resistance or dyslipidemia, preferably, the non-insulin dependent diabetes mellitus is type II diabetes.

[0045] The pharmaceutical composition of the present invention may be used in combination with one or more agents selected from metformin, thiazolidinediones, sulfonylureas, dipeptidyl peptidase inhibitors and sodium glucose cotransporters simultaneously, separately or successively.

[0046] Unlike other chemical products, even a trace amount of impurity components in drugs can cause risks in terms of safety, efficacy, quality controllability, etc. of clinical drug use. The present invention can obtain a pharmaceutical composition with good stability by controlling the components and the ratio of each component in the formulation without adding other auxiliary materials, effectively control the content of related substances, significantly reduce the content of specific impurities, greatly reduce the toxicity of the drug, and the increase in the content of specific impurities after long-term storage of the drug is slow, improving the safety of the drug. In addition, the filling is protected with nitrogen gas to improve the stability of the formulation. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0047] 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 of these should be included in the scope of patent protection defined in the claims of this application. It should be understood that the examples provided herein are only for illustrating the purpose of the present invention and should not be construed as limiting the present invention.

[0048] Example 1, [Table 1]

[0049] The water for injection is cooled to ≦25℃ and prepared for use. Weigh out 0.071g of the prescribed amount of disodium hydrogen phosphate and dissolve it in 90ml of water for injection, stir and dissolve, then add 0.1ml of 100g / L sodium hydroxide solution and stir evenly (0.2g of active ingredient is not added), weigh out 1.5g of the prescribed amount of propylene glycol and stir evenly, weigh out the prescribed amount of the active ingredient of the present invention and stir slowly to dissolve it, adjust the pH of the liquid to 7.5-8.0 with sodium hydroxide or hydrochloric acid solution, and use water for injection to make the liquid 100ml, and finally filter the liquid through a 0.22μm microporous filter, dispense into prefilled syringe bottles, fill with nitrogen gas, and then push in the plug. The samples were placed under conditions of 25°C±2°C / 60%RH±5%RH and 40°C, respectively, and their stability was examined. The examination indexes were appearance and related substances, and the results are shown in the table below.

[0050] [Table 2]

[0051] Example 2, [Table 3]

[0052] The injection water is cooled to ≦25℃ and prepared for use. Weigh out 0.071g of the prescribed amount of disodium hydrogen phosphate and dissolve in 90ml of injection water, stir and dissolve, then add 0.1ml of 100g / L sodium hydroxide solution and stir evenly, weigh out 1.5g of the prescribed amount of propylene glycol and stir evenly, weigh out 2.0g of the prescribed amount of the active ingredient of the present invention and stir slowly to dissolve it, adjust the pH of the liquid to 6.5, 7.0, 7.5, 7.8, 8.0, 8.5 respectively with sodium hydroxide or hydrochloric acid solution, and use injection water to make the above liquid constant to 100ml, finally filter the liquid through 0.22μm microporous filter, dispense into prefilled syringe bottle, fill with nitrogen gas, and then push in the plug. The sample was placed under the conditions of 25°C±2°C / 60%RH±5%RH and its stability was examined. The examination indexes were appearance and related substances. The results are shown in the table below.

[0053] [Table 4]

[0054] Example 3, [Table 5]

[0055] The injection water is cooled to ≦25℃ and prepared for use. Weigh out 0.071g of the prescribed amount of disodium hydrogen phosphate and dissolve it in 90ml of injection water, stir and dissolve, then add 100g / L sodium hydroxide solution of the prescribed amount, stir evenly, weigh out 1.5g of propylene glycol of the prescribed amount, stir evenly, measure the pH value of the solution, weigh out 2.0g of the active ingredient of the present invention of the prescribed amount, slowly add, stir for 45 minutes, observe the dissolution phenomenon of the active ingredient, measure the pH value of the solution, place the sample under the conditions of 25℃±2℃ / 60%RH±5%RH, 5℃±3℃, 40℃, and light irradiation, and consider its stability, carry out analysis and research on its impurities, consider the impact of different buffer concentrations on impurities, and provide a research concept for the selection of pharmaceutical compositions, and the research results are shown in the following table.

[0056] [Table 6]

[0057] Example 4, [Table 7]

[0058] The water for injection was cooled to ≦25℃ and prepared for use. The prescribed amount of disodium hydrogen phosphate was weighed and dissolved in 90ml of water for injection, stirred to dissolve, and then 0.1ml of 100g / L sodium hydroxide solution was added and stirred uniformly. The prescribed amount of propylene glycol was weighed and stirred uniformly. The prescribed amount of 2.0g of the active ingredient of the present invention was weighed and stirred slowly to dissolve it. The pH of the liquid was adjusted to 7.5-8.0 with sodium hydroxide or hydrochloric acid solution, respectively, and the above liquid was made into a constant volume of 100ml with water for injection. Finally, the liquid was filtered through a 0.22μm microporous filter, dispensed into a prefilled injection bottle, filled with nitrogen gas, and then plugged in. The sample was placed under the conditions of 25℃±2℃ / 60%RH±5%RH or 40℃, and its stability was examined. The examination indexes were appearance and related substances, and the results were shown in the table below.

[0059] [Table 8]

[0060] Example 5, [Table 9]

[0061] The water for injection was cooled to ≦25℃ and prepared for use. The prescribed amount of disodium hydrogen phosphate 0.071g was weighed and dissolved in 90ml of water for injection, stirred to dissolve, and then 0.1ml of 100g / L sodium hydroxide solution was added and stirred uniformly. The prescribed amount of osmotic pressure regulator was weighed and stirred uniformly. The prescribed amount of active ingredient of the present invention 2.0g was weighed and stirred slowly to dissolve it. The pH of the liquid was adjusted to 7.5-8.0 with sodium hydroxide or hydrochloric acid solution, respectively. The liquid was adjusted to 100ml with water for injection. Finally, the liquid was filtered through a 0.22μm microporous filter and dispensed into a prefilled injection bottle, filled with nitrogen gas, and then plugged in. The sample was placed under the conditions of 25℃±2℃ / 60%RH±5%RH, and its stability was examined. The examination indexes were appearance and related substances, and the results were shown in the table below.

[0062] [Table 10]

[0063] Example 6, [Table 11]

[0064] The water for injection was cooled to ≦25℃ and prepared for use. The prescribed amount of disodium hydrogen phosphate was weighed and dissolved in 90ml of water for injection, stirred and dissolved, and then 0.1ml of 100g / L sodium hydroxide solution was added and stirred evenly. The prescribed amount of propylene glycol was weighed and stirred evenly. The prescribed amount of the active ingredient of the present invention was weighed and dissolved by slowly stirring. The pH of the liquid was adjusted to 7.5-8.0 with sodium hydroxide or hydrochloric acid solution, respectively. The liquid was adjusted to 100ml with water for injection. Finally, the liquid was filtered through a 0.22μm microporous filter, dispensed into prefilled injections, and plugs were inserted. The sample was placed under conditions of 25℃±2℃ / 60%RH±5%RH, 5℃±3℃, 40℃, light irradiation, and low temperature cycle, and its stability was examined. The examination indexes were appearance and related substances, and the results were shown in the table below.

[0065] [Table 12]

[0066] Test Example 1 Hemolysis test 1. Subject: Test product: Injection solution prepared in Example 1 (active ingredient 2.0 g), 1.2 Solvent: Sodium Chloride Injection.

[0067] 2. Experimental Method: Different volumes (0.5-0.1mL) of test samples and different volumes (2.0-2.4mL) of sodium chloride injection were added to glass test tubes containing 2.5mL of 2% rabbit red blood cell suspension, and sodium chloride injection and sterile water for injection were used as negative and positive controls, respectively. The total volume of each test tube was 5.0mL, and the tubes were incubated in an electric thermostatic box (set temperature 37℃) for 3 hours to observe the lysis and aggregation of red blood cells.

[0068] In the test tubes containing 0.5-0.1 mL of the test product and the negative control product, it was found that the red blood cells sank to the bottom of the tube, the upper layer solution was colorless and transparent, the red blood cells at the bottom of the tube were uniformly dispersed after shaking, and there was no hemolysis or agglutination. In the sterile water for injection positive control tube, it was found that some red blood cells sank to the bottom of the tube, the upper layer solution was transparent red, the red blood cells at the bottom of the tube were uniformly dispersed after shaking, there was no partial hemolysis, and there was no agglutination.

[0069] Under the test conditions, the pharmaceutical composition injection of the present invention having a labeling concentration of 20 mg / mL did not cause hemolysis of rabbit red blood cells in a 0.1-0.5 mL test system, nor did it cause red blood cell aggregation.

[0070] Test Example 2: Subcutaneous irritation study 1. Subject: Test product: Injection solution prepared in Example 1 (active ingredient 2.0 g),

[0071] 2. Experimental Method: In the test, New Zealand rabbits were selected, and the same-body left-right self-control method was used to administer the drug subcutaneously a total of four times to the New Zealand rabbits. During the period in which the irritation reaction caused by local subcutaneous administration was tested, no abnormalities related to the injection of the pharmaceutical composition of the present invention were observed in any of the animals after local administration.

[0072] Test Example 3: Study of the therapeutic effect of the pharmaceutical composition of the present application after a single subcutaneous injection into type 2 diabetic db / db mice 1. Subject: Test product: Injection solution prepared in Example 1 (active ingredient 2.0 g), 2. Experimental Method After a single administration, the effects on blood glucose, serum insulin level, body weight and food intake of type 2 diabetes db / db mice were observed, and the duration of efficacy after a single administration was also observed. The experimental results showed that the pharmaceutical composition can significantly lower random blood glucose in db / db mice after a single subcutaneous injection at doses of 1, 3 and 10 nmol / kg, exhibiting a good dose-effect relationship, and a clear random blood glucose lowering effect can be seen at a dose of 3 nmol / kg. The duration of hypoglycemic effect after a single subcutaneous injection of the pharmaceutical composition is dose-related, with the random blood glucose lowering effect being maintained for 1 day at 1 nmol / kg, and 2-3 days at 3 nmol / kg and 10 nmol / kg.

[0073] [Table 13]

[0074] In addition, Mouse Insulin ELISA Kit was used to measure the serum insulin content of each group of db / db mice 2 hours after administration. The results showed that after a single subcutaneous injection of 1, 3, 10 nmol / kg of this pharmaceutical composition into db / db mice, the serum insulin levels of the mice in each dose group were significantly increased compared with the vehicle group, showing a good dose-effect relationship, and there was a positive correlation between the increase in serum insulin and the decrease in blood glucose.

[0075] [Table 14]

[0076] Test Example 4: Study of the therapeutic effect of the pharmaceutical composition of the present application after long-term subcutaneous injection into type 2 diabetic db / db mice 1. Subject: Test product: Injection solution prepared in Example 1 (active ingredient 2.0 g),

[0077] 2. Experimental Method: Type 2 diabetic db / db mice were administered once every 3 days for 9 consecutive doses. After the end of the study (d26), the liver weights of db / db mice in the 1, 3, and 10 nmol / kg groups were 2.313±0.113g, 2.267±0.108g, and 1.993±0.104g, respectively, all of which were significantly lower than the vehicle group (p<0.05, p<0.05, p<0.001), and there was no statistically significant difference in pancreas weight compared to the vehicle group (p>0.05).

[0078] In summary, when this pharmaceutical composition is administered repeatedly for a long period of time in a dose-dependent manner once every three days, it can reduce the body weight and liver weight of type 2 diabetic db / db mice, and at doses of 3 and 10 nmol / kg, it has obvious random hypoglycemic effect.

[0079] Test Example 5: Pharmacokinetic study of a single subcutaneous or intravenous injection of the pharmaceutical composition of the present application in cynomolgus monkeys 1. Subject: The injection solution prepared in Example 3,

[0080] 2. Experimental Method: The doses administered in each of the four groups, i.e., the intravenous group, the low-dose subcutaneous injection group, the medium-dose subcutaneous injection group, and the 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.

[0081] After intravenous injection, the elimination half-life t1 / 2 of the drug of the present invention in the cynomolgus monkey was between 50.7 and 51.5 h, and the drug was slowly eliminated; the elimination rate CL was between 0.89 and 0.93 mL / h / kg, and the amount of elimination was low; the apparent distribution volume Vd was between 66.0 and 68.1 mL / kg, and the drug was mainly distributed in plasma; after a single subcutaneous injection, the time to reach the peak plasma concentration Tmax of the drug of the present invention in the cynomolgus monkey was 13.3 to 18.0 h, and the plasma elimination half-life T1 / 2 was between 48.9 and 65.4 h, and the drug was slowly eliminated; the exposure amount (AUClast) of the drug of the present invention in the animals of different sexes was not statistically different (P>0.05). Within the dose range of 0.02 to 0.2 mg / kg, the exposure amount (AUC last ) increased with increasing dose, and the rate of increase in exposure was close to that of increase in dose.

Claims

1. A stable GIP / GLP-1 dual receptor agonist pharmaceutical composition comprising an active ingredient, a buffer, an osmolality adjusting agent, and a pH adjusting agent, wherein the active ingredient has the structure: 【Chemistry 1】 13. The pharmaceutical composition according to claim 12,

2. The pharmaceutical composition according to claim 1, characterized in that the pharmaceutical composition is an injection solution, and the concentration of the active ingredient is 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, characterized in that it contains 0.05% to 4.0% (W / V) of active ingredient, preferably 0.1% to 3.0% (W / V).

4. 2. The pharmaceutical composition according to claim 1, characterized in that the buffer is selected from phosphate buffer, acetate buffer, citrate buffer, carbonate buffer, tartrate buffer, Tris buffer, histidine salt, preferably citrate buffer or phosphate buffer, more preferably disodium hydrogen phosphate.

5. 2. The pharmaceutical composition according to claim 1, characterized in that it contains 0.05% to 3.0% (W / V) of a buffer solution, preferably 0.05% to 2.0% (W / V), and more preferably 0.05% to 1.0% (W / V).

6. 2. The pharmaceutical composition according to claim 1, wherein the osmolality adjusting agent is one or more selected from mannitol, lactose, sucrose, propylene glycol, glycerin, preferably propylene glycol or mannitol.

7. The pharmaceutical composition according to claim 1, characterized in that the injection solution contains an osmolality adjusting agent at 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. 2. The pharmaceutical composition according to claim 1, wherein the pH adjusting agent is one or more selected from hydrochloric acid and sodium hydroxide.

9. It contains 0.1% to 3.0% (W / V) of an active ingredient, 0.05% to 3.0% (W / V) of a buffer solution, 0.5% to 5.0% (W / V) of an osmotic pressure adjusting agent, and an appropriate amount of a pH adjusting agent; Preferably, the composition contains 0.1% to 3.0% (W / V) of an active ingredient, 0.05% to 2.0% (W / V) of a buffer solution, 1.0% to 3.0% (W / V) of an osmotic pressure adjusting agent, and an appropriate amount of a pH adjusting agent; More preferably, the composition contains 0.1% to 3.0% (W / V) of an active ingredient, 0.05% to 1.0% (W / V) of a buffer solution, 1.0% to 2.0% (W / V) of an osmotic pressure adjusting agent, and an appropriate amount of a pH adjusting agent; More preferably, the pharmaceutical composition according to claim 1 contains 0.1% to 3.0% (W / V) of an 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.

10. The pharmaceutical composition according to claim 1, characterized in that the weight ratio of the active ingredient to the buffer solution is 1:0.01-10, preferably 1:0.02-1, and more preferably 1:0.02-0.

5.

11. The pharmaceutical composition according to claim 1, characterized in that the weight ratio of the active ingredient to the osmotic pressure adjusting agent is 1:0.1-20, preferably 1:0.4-10.

12. The pharmaceutical composition according to claim 9, wherein the buffer solution 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. 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. The pharmaceutical composition according to claim 1, characterized in that the pH range of the pharmaceutical composition is 6.5 to 9.0, preferably 7.0 to 8.5, and more preferably 7.0 to 8.

0.

15. 15. The pharmaceutical composition according to claim 1, further comprising a preservative, said preservative being one or more selected from m-cresol, phenol, phenylcarbinol, phenylethanol, parahydroxybenzoic acid esters, hydroxybenzoic acid esters, benzyl alcohol, chlorobutanol, phenoxyethanol and methylparaben.

16. A method for producing the pharmaceutical composition according to claims 1 to 15, comprising dissolving a buffer and an osmolality adjusting agent in water for injection, dissolving an active ingredient in said liquid drug, stirring to dissolve, adding a pH adjusting agent, adjusting the volume to volume, filtering and dispensing.

17. The use of the pharmaceutical composition according to claims 1 to 15 in the manufacture of a medicament for treating non-insulin-dependent diabetes mellitus, insulin-dependent diabetes mellitus, obesity, insulin resistance or dyslipidemia, preferably, the non-insulin-dependent diabetes mellitus is type II diabetes.

Images