TWI494313B - Pharmaceutical composition for the treatment of type 2 diabetes in mammals including human beings - Google Patents

Description

Treatment of medical compositions including type 2 diabetes in human mammals

The present invention relates to a pharmaceutical composition for treating type 2 diabetes in a mammal including a human, particularly comprising rosiglitazone or pioglitazone or a pharmaceutically acceptable salt thereof and (R)-7-[3-amino-4 -(2,4,5-trifluoro-phenyl)-butenyl]-3-trifluoromethyl-5,6,7,8-tetrahydro-imidazo[1,5-a]pyrazine-1- A pharmaceutical composition of a fixed dose combination of a methyl carboxylate or a pharmaceutically acceptable salt thereof, a method of preparing the pharmaceutical composition, and the use of the pharmaceutical composition for treating type 2 diabetes in a mammal including a human.

Human type 2 diabetes is a chronic and progressive disease caused by the complex pathophysiology of dual endocrine defects involving insulin resistance and weakened insulin secretion. The treatment of human type 2 diabetes generally begins with diet and exercise followed by oral monotherapy for antidiabetic drugs. For many patients, this regimen does not adequately control blood glucose during long-term treatment, which results in the need for combination therapy within a few years after diagnosis. However, co-prescription of two or more oral anti-diabetic drugs can lead to complex and difficult treatment options for many patients used. Combining two or more anti-diabetic agents into a single formulation provides a possible means of delivering combination therapies that do not increase the complexity of the patient's daily treatment regime.

Dipeptidyl peptidase-IV (DPP-IV) inhibitors are new agents developed to treat or improve glycemic control in patients with type 2 diabetes. The drugs used to treat human type 2 diabetes in current clinical trials include MK-0431, vildagliptin (LAF-237), saxagliptin (BMS-47718), P93/01 (Prosidon), SYR322 (Takeda), GSK823093. , Roche0730699, TS021 (Taisho), E3024 (Eisai) and PHX-1149 (Phenomix). For example, oral administration of vildagliptin to human type 2 diabetic patients has been found to reduce fasting glucose and postprandial glucose deviation associated with significantly reduced HbAIC levels. For a review of the use of DDP-IV in the treatment of type 2 diabetes, see the following publications: (1) H.-U. Demuth et al., "Type 2diabetes-Theraphy with dipeptidyl peptidase IV inhibitors", Biochim. Biophvs. Acta. 1751:33 -44 (2005) and (2) K. Augustyns et al., "Inhibitors of proline-specific dipeptidyl peptidases: DPP4 inhibitors as a novel approach for the treatment of Type 2 diabetes", Expert Opin. Ther. Patants, 15:1387- 1407 (2005).

At present, some DPP-IV inhibitors have been disclosed (US 5,462,928, US Pat. No. 5,543,396, WO 9515309, WO2003004498, WO2003082817, WO2004032836, WO2004085661), wherein the DPP-IV inhibitor MK-0431 produced by Merck Company shows good DPP-IV inhibitory activity and selection. Sex, and was listed in 2006.

In order to administer a PPAR anti-diabetic agent, particularly glitazone, to an adult diabetic patient (body weight: 50 kg), for example, the daily dose is usually 0.01 to 1000 mg, preferably 0.1 to 500 mg. This dose is administered once to several times a day. In particular, when pioglitazone hydrochloride is used as the insulin sensitizer, the daily dose of pioglitazone hydrochloride is usually 7.5 to 60 mg, preferably 15 to 45 mg. When rosiglitazone (or maleate) is used as the insulin sensitizer, the dose of rosiglitazone per day is usually from 1 to 12 mg, preferably from 2 to 12 mg.

The glitazone is preferably pioglitazone, pioglitazone hydrochloride or rosiglitazone, pioglitazone maleate or rosiglitazone, and particularly preferably pioglitazone hydrochloride.

Pharmaceutical dosage forms of combinations of antidiabetic agents and thiazolidinedione derivatives have been proposed in the art. For example, EP 0 749 751 teaches a pharmaceutical composition comprising an insulin sensitivity enhancer which is a thiazolidinedione compound in combination with other anti-diabetic agents. More particularly, EP 0 749 751 teaches that a preferred insulin sensitivity enhancer is pioglitazone, which can be combined with other anti-diabetic agents such as rosiglitazone, phenformin or buformin, and these drugs can be combined (mixed and/or coated) Conventional excipients to provide taste masking or sustained release. Another example of a combination of an antidiabetic agent and a thiazolidinedione derivative is U.S. Patent No. 6,011,049, which teaches a sustained release form, such as an osmotic pump or a skin patch containing pioglitazone or troglitazone. And a single pharmaceutical composition of rosiglitazone or pioglitazone. Other combinations of anti-diabetic agents and thiazolidinedione derivatives can be found in U.S. Patent Nos. 6,524,621; 6,475, 521; 6, 451, 342 and 6, 153, 632, and PCT patent application WO 0 1/3 594, incorporated herein by reference.

Has the following structural formula (R)-7-[3-amino-4-(2,4,5-trifluoro-phenyl)-butenyl]-3-trifluoromethyl-5,6,7,8- The tetrahydro-imidazo[1,5-a]pyrazine-1-carboxylic acid methyl ester is a compound of the formula (I), and the compound and its salt are disclosed in Chinese Patent Application No. 200710302335.9.

The present invention provides a pharmaceutical composition for treating type 2 diabetes in a mammal comprising a human, the pharmaceutical composition comprising a therapeutically effective amount of rosiglitazone or pioglitazone or a pharmaceutically acceptable salt thereof and a compound of formula (I) Or a pharmaceutically acceptable salt thereof, wherein the weight ratio of rosiglitazone or a pharmaceutically acceptable salt thereof to the compound of the formula (I) or a pharmaceutically acceptable salt thereof is 1:6.25 to 1:75, pioglitazone or The weight ratio of the pharmaceutically acceptable salt thereof to the compound of the formula (I) or a pharmaceutically acceptable salt thereof is from 1:3 to 1:50.

The two active pharmaceutical ingredients in the pharmaceutical compositions of the invention are either immediate or slow release. The pharmaceutical composition of the present invention may be in the form of a tablet, and particularly a film-coated tablet, and may also be other oral dosage forms such as capsules (including hard capsules and soft capsules), oral solutions, sustained release agents, pills. , granules, granules, sustained release pellets, etc.

One aspect of the invention relates to a dosage form for the pharmaceutical administration of a fixed dose combination of a compound of formula (I) or a pharmaceutically acceptable salt thereof and rosiglitazone or pioglitazone or a pharmaceutically acceptable salt thereof. The dosage form can be in powder or solid form and includes tablets, capsules, sachets and the like. Particular solid dosage forms relate to tablets containing a fixed dose combination of a compound of formula (I) or a pharmaceutically acceptable salt thereof and rosiglitazone or pioglitazone or a pharmaceutically acceptable salt thereof.

The present invention also provides a process for the preparation of a pharmaceutical composition of a fixed dose combination of a compound of formula (I) or a salt thereof and rosiglitazone or pioglitazone or a salt thereof by a dry or wet treatment method. Dry processing methods include dry compression and dry granulation, and wet processing methods include wet granulation.

Another aspect of the invention provides the use of a pharmaceutical composition of the invention in the treatment of Type 2 diabetes in a mammal comprising a human comprising administering to the subject in need of such treatment a therapeutically effective amount of a pharmaceutical composition of the invention.

In a particular embodiment of the invention, the pharmaceutical composition comprises (1) a compound of formula (I) or a pharmaceutically acceptable salt thereof, which is the first active pharmaceutical ingredient; (2) rosiglitazone or pioglitazone or a a salt, which is a second active pharmaceutical ingredient; and (3) a lubricant or a glidant. In a particular embodiment of this aspect of the invention, the pharmaceutical composition may also contain one or more excipients selected from one or more binders (binding agents); one or more diluents; one or more a surfactant or wetting agent; one or more disintegrants; and one or more antioxidants.

The pharmaceutically acceptable salts of the compounds of formula (I), rosiglitazone or pioglitazone include, but are not limited to, phosphates, hydrochlorides, sulfates, nitrates, hydrobromides, methanesulfonates, Malay Acid salt, tartrate, succinate, acetate, trifluoroacetate, fumarate, citrate, citrate, besylate, benzoate, naphthalene sulfonate, lactate or Malate.

The compound of the formula (I) or a salt thereof incorporated in the pharmaceutical composition of the present invention has a daily dose of 25 mg to 300 mg per person. The daily dose per person of the compound of the formula (I) or a salt thereof is preferably from 50 mg to 200 mg. Individual individual doses per day are 25, 50, 75, 100, 150, 200, 250 and 300 mg of the compound of formula (I) or a salt thereof.

The daily dose of rosiglitazone or a salt thereof incorporated in the fixed dose combination of the present invention is from 1 mg to 12 mg per day, and the individual daily doses are 1, 2, 4, 8, 10 and 12 mg per person, The daily dose of pioglitazone or its salt is 7.5 mg to 60 mg per person, and the individual daily doses are 7.5, 15, 30, 45 and 60 mg per person. These daily doses of rosiglitazone or pioglitazone or their salts represent daily doses approved for commercial treatment of human type 2 diabetes in China and/or the United States.

In the fixed dose combination of the present invention, the specific embodiment of the daily dose of the compound of the formula (I) or a salt thereof and rosiglitazone or pioglitazone or a salt thereof is as follows:

a compound of the formula (I) or a salt thereof (mg) 25, 50, 75, 100, 150, 200, 250, 300

Rosiglitazone or its salt (mg) 1,2,4,8,12

Pioglitazone or its salt (mg) 7.5, 15, 30, 45, 60

It may be any therapeutically effective amount of a compound of formula (I) or a salt thereof, in combination with any therapeutically effective amount of rosiglitazone or pioglitazone or a salt thereof, for example: 50+2, 50+4, 100+2, 100+ 4 or 50+7.5, 50+15, 50+30, 50+45, 75+7.5, 75+15, 100+30, 100+45 and so on.

The two active pharmaceutical ingredients in the pharmaceutical composition of the present invention can be in four release forms:

The pharmaceutical composition of the present invention is prepared by a wet method or a dry method. In one embodiment, the pharmaceutical composition is prepared by a wet processing method. In one class of this embodiment, the pharmaceutical composition is prepared by a wet granulation process. In the wet granulation, high shear granulation or fluidized bed granulation can be applied. In one embodiment, the use of fluidized bed granulation has the advantage of making the tablet have a higher radial strength.

The pharmaceutical composition obtained by dry or wet processing can be compressed into tablets, packaged or metered into sachets.

The pharmaceutical composition contains one or more lubricants or glidants. Examples of the lubricant include magnesium stearate, calcium stearate, stearic acid, sodium stearyl fumarate, hydrogenated castor oil or a mixture thereof. A preferred lubricant is magnesium stearate or sodium stearyl fumarate or a mixture thereof. Examples of the glidant include colloidal cerium oxide, calcium phosphate, magnesium citrate, and talc.

The pharmaceutical compositions of the present invention optionally contain one or more binders. Embodiments of the binder include hydroxypropyl cellulose (HPC), hydroxypropyl methylcellulose (HMPC), hydroxyethyl cellulose, starch 1500, polyvinylpyrrolidone (polypyrrolidone), and copolyenolone. . A preferred binder is polyvinylpyrrolidone.

The pharmaceutical compositions of the present invention may also optionally contain one or more diluents. Examples of the diluent include mannitol, sorbitol, calcium dihydrogen phosphate dihydrate, microcrystalline cellulose, and powdered cellulose. A preferred diluent is microcrystalline cellulose. Microcrystalline cellulose can be obtained from several suppliers including Avicel PH 101, Avicel PH 102, Avicel PH 103, Avicel PH 105 and Avicel PH 200 manufactured by FMC Corporation.

The pharmaceutical composition of the present invention may further optionally contain a disintegrating agent. The disintegrant may be one of several modified starches, modified cellulose polymers or polycarboxylic acids, such as crosslinked sodium carboxymethylcellulose, sodium starch glycolate, potassium bolconlin and hydroxymethylcellulose. Calcium (CMC Calcium). In one embodiment, the disintegrant is crosslinked hydroxymethylcellulose sodium. Crosslinked hydroxymethylcellulose sodium NF type A is commercially available under the trade designation "Ac-di-sol".

The pharmaceutical compositions of the present invention may also optionally contain one or more surfactants or wetting agents. The surfactant can be an anionic, cationic or neutral surfactant. Anionic surfactants include sodium lauryl sulfate, sodium dodecyl sulfate, sodium oleyl sulfate, and sodium laurate mixed with stearic acid and talc. Cationic surfactants include benzalkonium chloride and alkyltrimethylammonium bromide. Neutral surfactants include glycerol monooleate, polyoxyethylene sorbitan fatty acid esters, polyvinyl alcohol and sorbitan esters. Embodiments of the wetting agent include poloxamers, polyoxyethylene alkyl ethers, polyoxyethylene castor oil derivatives, and polyoxyethylene stearates.

An antioxidant may optionally be added to the formulation to give it chemical stability. The antioxidant is selected from the group consisting of α-tocopherol, γ-tocopherol, δ-tocopherol, tocopherol enriched natural extract, L-ascorbic acid and its sodium or calcium salt, ascorbate palmitate, citric acid Propyl ester, octyl phthalate, dodecyl decanoate, butylated hydroxytoluene (BHT) and butylated hydroxyanisole (BHA). In one embodiment, the antioxidant is BHT or BHA.

A preferred dosage form of the pharmaceutical composition of the present invention is a tablet prepared by a compression method. The tablet may be coated with a mixture of, for example, hydroxypropylcellulose and hydroxypropylmethylcellulose, which contains titanium dioxide and/or other colorants such as iron oxide, dyes and lakes; polyvinyl alcohol ( A mixture of PVA) and polyethylene glycol (PEG) containing titanium dioxide and/or other colorants such as iron oxide, dyes and lakes; or any other suitable immediate release coating. The coating provides taste masking and additional stability to the final tablet. Commercially available coatings are supplied by Colorcon as Opadry ^® for formulating powder mixtures.

Finally, sweeteners and/or flavoring agents can be added if desired.

The rosiglitazone or pioglitazone or a salt thereof of the present invention may be either immediate release or slow release, and the compound of the formula (I) or a salt thereof may be either immediate release or slow release.

The pharmaceutical tablet compositions of the present invention may also contain one or more additional formulation ingredients selected from excipients known in the art of various pharmaceutical formulations. Depending on the desired properties of the pharmaceutical combination, any of the ingredients may be selected individually or in combination based on their known use in preparing the tablet composition. Such ingredients include, but are not limited to, diluents, compression aids, glidants, disintegrants, lubricants, perfumes, flavoring agents, sweeteners, and preservatives.

The term "tablet" as used herein is intended to include compressed drug dosage formulations of all shapes and sizes, whether coated or not. Substances that can be used for coating include hydroxypropylcellulose, hydroxypropylmethylcellulose, titanium dioxide, talc, sweeteners, colorants, and flavoring agents.

The invention also provides a method of treating a mammal, including human type 2 diabetes, by orally administering to a subject in need of such treatment a therapeutically effective amount of a fixed dose combination pharmaceutical composition of the invention. In one embodiment, the subject in need of such treatment is a human. In another embodiment, the pharmaceutical composition is in the form of a tablet, which may also be in the form of a capsule.

The pharmaceutical composition containing the fixed dose combination can be administered once daily (QD), twice daily (BID) or three times daily (TID).

The compound of the formula (I) or a salt thereof has a longer inhibition time for DPP-IV than MK-0431 and a greater inhibitory strength than MK-0431. Through experimental research, the present inventors have surprisingly found that the compound effect of the compound of the formula (I) or a salt thereof and rosiglitazone or pioglitazone or a salt thereof is remarkably higher than that of any of the components alone, especially when Rogge column The compounding ratio of the ketone or a salt thereof and the compound of the formula (I) or a salt thereof is from 1:6.25 to 1:75, and the compounding ratio of the pioglitazone or a salt thereof and the compound of the formula (I) or a salt thereof is from 1:3 to 1:50. better result. Thus, a composition of the compound of the formula (I) or a salt thereof and rosiglitazone or a salt thereof, a compound of the formula (I) (hereinafter referred to as compound A, code SP2086) or a salt thereof, and a combination of pioglitazone or a salt thereof It is of great clinical significance.

The following examples further describe and illustrate embodiments within the scope of the invention. The examples and the test examples are given for the purpose of illustration only, and are not intended to be construed as limiting the scope of the invention.

Example 1. Compound A phosphate and rosiglitazone maleate compound tablet

Prescription Each tablet contains:

Compound A Phosphate 50 mg

Rosiglitazone maleate 4 mg

Microcrystalline cellulose 12 mg

2% starch slurry

Magnesium stearate 0.5 mg

Preparation method: Compound A phosphate, rosiglitazone maleate, microcrystalline cellulose were mixed and wet granulated with 2% starch slurry. Dry, add magnesium stearate, and mix and compress.

Example 2, Compound A Phosphate and Pioglitazone Hydrochloride Compound Tablets

Prescription Each tablet contains:

Compound A Phosphate 50 mg

Pioglitazone 15 mg

Microcrystalline cellulose 12 mg

2% starch slurry

Magnesium stearate 0.5 mg

Preparation method: Compound A phosphate, pioglitazone hydrochloride, microcrystalline cellulose were mixed and wet granulated with 2% starch slurry. Dry, add magnesium stearate, and mix and compress.

Example 3, Compound A Hydrochloride and Rosiglitazone Maleate Compound Tablets

Prescription Each tablet contains:

Compound A Hydrochloride 50 mg

Rosiglitazone maleate 15 mg

Microcrystalline cellulose 12 mg

2% starch slurry

Magnesium stearate 0.5 mg

Preparation method: Compound A hydrochloride, rosiglitazone maleate, microcrystalline cellulose was mixed and wet granulated with 2% starch slurry. Dry, add magnesium stearate, and mix and compress.

Example 4, Compound A Hydrochloride and Pioglitazone Hydrochloride Compound Tablets

Prescription Each tablet contains:

Compound A Hydrochloride 50 mg

Pioglitazone 15 mg

Microcrystalline cellulose 12 mg

2% starch slurry

Magnesium stearate 0.5 mg

Preparation method: Compound A hydrochloride, pioglitazone hydrochloride, microcrystalline cellulose was mixed and wet granulated with 2% starch slurry. Dry, add magnesium stearate, and mix and compress.

Example 5, Compound A mesylate salt and rosiglitazone maleate compound tablet

Prescription Each tablet contains:

Compound A Mesylate 50 mg

Rosiglitazone maleate 15 mg

Microcrystalline cellulose 12 mg

2% starch slurry

Magnesium stearate 0.5 mg

Preparation method: Compound A methanesulfonate, rosiglitazone maleate, microcrystalline cellulose was mixed and wet granulated with 2% starch slurry. Dry, add magnesium stearate, and mix and compress.

Example 6, Compound A mesylate salt and pioglitazone hydrochloride compound tablet

Prescription Each tablet contains:

Compound A Mesylate 50 mg

Pioglitazone 15 mg

Microcrystalline cellulose 12 mg

2% starch slurry

Magnesium stearate 0.5 mg

Preparation method: Compound A mesylate, pioglitazone hydrochloride, microcrystalline cellulose was mixed and wet granulated with 2% starch slurry. Dry, add magnesium stearate, and mix and compress.

Example 7, Compound A Tartrate and Rosiglitazone Maleate Compound Tablets

Prescription Each tablet contains:

Compound A Tartrate 50 mg

Rosiglitazone maleate 15 mg

Microcrystalline cellulose 12 mg

2% starch slurry

Magnesium stearate 0.5 mg

Preparation method: Compound A tartrate, rosiglitazone maleate, microcrystalline cellulose was mixed and wet granulated with 2% starch slurry. Dry, add magnesium stearate, and mix and compress.

Example 8, Compound A Tartrate and Pioglitazone Hydrochloride Compound Tablets

Prescription Each tablet contains:

Compound A Tartrate 50 mg

Pioglitazone 15 mg

Microcrystalline cellulose 12 mg

2% starch slurry

Magnesium stearate 0.5 mg

Preparation method: Compound A tartrate, pioglitazone hydrochloride, microcrystalline cellulose was mixed and wet granulated with 2% starch slurry. Dry, add magnesium stearate, and mix and compress.

Example 9, Compound A Malate and Rosiglitazone Maleate Compound Tablets

Prescription Each tablet contains:

Compound A Malate 50 mg

Rosiglitazone maleate 15 mg

Microcrystalline cellulose 12 mg

2% starch slurry

Magnesium stearate 0.5 mg

Preparation method: Compound A malate, rosiglitazone maleate, microcrystalline cellulose was mixed and wet granulated with 2% starch slurry. Dry, add magnesium stearate, and mix and compress.

Example 10, Compound A Malate and Pioglitazone Hydrochloride Compound Tablets

Prescription Each tablet contains:

Compound A Malate 50 mg

Pioglitazone 15 mg

Microcrystalline cellulose 12 mg

2% starch slurry

Magnesium stearate 0.5 mg

Preparation method: Compound A malate, pioglitazone hydrochloride, microcrystalline cellulose was mixed and wet granulated with 2% starch slurry. Dry, add magnesium stearate, and mix and compress.

Similarly, a compound containing different ratios of two active ingredients, such as rosiglitazone or a salt thereof, and Compound A or a salt thereof, can be prepared by adjusting the amount of Compound A or a salt thereof, rosiglitazone or a salt thereof in the compound. The ratio is 1:6.25, 1:12.5, 1:25, 1:50, 1:75, and so on.

Alternatively, a compound containing two active ingredients in different ratios, for example, a ratio of pioglitazone or a salt thereof to Compound A or a salt thereof, of 7.5:25, is prepared by adjusting the amount of Compound A or a salt thereof, pioglitazone or a salt thereof in the compound. 7.5:50, 7.5:75, 7.5:100, 7.5:150, 7.5:375, etc.

Test Example 1: In vitro activity and selectivity of Compound A and MK-0431:

Thaw DPP-IV-Glo., buffer and equilibrate to room temperature before use. Pre-buffer frozen luciferin detection reagent, suspend DPP-IV-Glo. Add ultrapure water to the matrix and mix gently. In a 1 mM matrix, place the luciferin assay reagent in a brown bottle and add DPP-IV-Glo. The luciferin test reagent should be dissolved within 1 minute, dissolve the test compound in DMSO to 50 times the final operating concentration, add 50 μl of test compound 2 μL to each tube, and add 2 μL DMSO to the negative control and blank control. Add 46 μL of Tris buffer to each tube, add 48 μL of Tris buffer to the blank, add 2 μL of LDPP-IV enzyme to each tube of the negative control and test sample, mix by shaking and centrifuge the tube. The contents of the tubes were all transferred to 96-well plates at a ratio of 1:49 for the mixed matrix and DPP-IV-Glo. Mix the vibrations to mix thoroughly. Allow to stand at room temperature for 30 to 60 minutes before use, add 50 μL of DPP-IV-Glo. and substrate mixture to each 96-well plate well, seal the plate with a sealing film, and use a plate shaker at 300 The 96-well material was slowly mixed at 500 rpm/30 s. Incubate for 30 minutes to 3 hours at room temperature and measure the chemiluminescence count on a NOVOstar multi-function microplate reader.

Results: The inhibitory activity of Compound A on DPP-IV was better than that of the control drug MK-0431, and the selectivity was also stronger than MK-0431.

Test Example 2: Oral Glucose Tolerance Study of Different Compositions of Rosiglitazone and Compound A Pharmaceutical Compositions in Normal ICR Mice

Male ICR mice were orally administered with double distilled water after 6 hours of fasting, and different ratios of rosiglitazone maleate: Compound A phosphate = 1:6.25, 1:12.5, 1:25, 1:50, 1: 75 The same dose (10 mg/kg) of the pharmaceutical composition, each group was orally administered with glucose 2.5 g/kg at 30 minutes for oral glucose tolerance test. Blood was collected at 0, 30, 60, and 120 minutes after the administration of the sugar, and the serum glucose concentration was measured.

Serum glucose determination method;

Glucose kit was used to determine the glucose content in the serum. 250 μl of the enzyme working solution was added, 5 μl of serum was added, and a blank tube (add 5 μl of double distilled water) and a standard tube (add 5 μl of glucose standard solution) were added, and the mixture was mixed, and the water bath was heated at 37 ° C for 20 minutes. Zero with a blank tube and colorimetric at OD505nm.

Serum glucose content BG (nmol / l) = OD _{sample tube} / OD _{standard tube} × 5.55

Data processing and statistical analysis;

1. Statistical analysis of data using mean ± standard deviation (Mean ± SD) and Student-t test

2. Calculate the percentage of blood glucose drop at 30 minutes after feeding sugar and the area under the curve AUC

Test Example 3: Oral glucose tolerance study of different ratios of pioglitazone and Compound A pharmaceutical composition in normal ICR mice

Male ICR mice were orally administered with double distilled water after 6 hours of fasting, with different ratios of formulation (pioglitazone hydrochloride: compound A phosphate = 7.5:25, 7.5:50, 7.5:75, 7.5:100, 7.5:150, 7.5: 375) A mixture of the same dose (30 mg/kg), each group was orally administered with glucose 2.5 g/kg at 30 minutes for oral glucose tolerance test. Blood was collected at 0, 30, 60, and 120 minutes after the administration of the sugar, and the serum glucose concentration was measured. Serum glucose determination method; glucose content in serum was determined by glucose kit, 250 μl enzyme working solution was taken, 5 μl serum was added, blank tube (add 5 μl double distilled water) and standard tube (add 5 μl glucose standard solution), and mixed. The water bath was incubated at 37 ° C for 20 minutes, and the blank tube was adjusted to zero, and the color was measured at OD 505 nm.

Serum glucose content BG (nmol / l) = OD sample tube / OD standard tube × 5.55

Data processing and statistical analysis;

1. Statistical analysis of data using mean ± standard deviation and Student-t test

2. Calculate the percentage of blood glucose drop at 30 minutes after feeding sugar and the area under the curve AUC

Test Example 4: Effect of Compound A Phosphate and MK-0431 in Combination with Rosiglitazone Maleate or Pioglitazone Hydrochloride in Wistar Rats with Genetic Obesity and Diabetes Mellitus

Male Wistar rats aged 14 to 19 weeks were divided into 9 groups of 5 to 6 each, taking distilled water, Compound A phosphate, rosiglitazone maleate, pioglitazone hydrochloride, MK-0431, MK. -0431 + rosiglitazone maleate, MK-0431 + pioglitazone hydrochloride, compound A phosphate + rosiglitazone maleate and compound A phosphate + pioglitazone hydrochloride for 14 days. Blood was collected from the tail vein, and plasma glucose and hemoglobin A1 were separately determined enzymatically using a commercial kit (NC-ROPET, Nippon Chemiphar CO.). The results were expressed as mean ± standard deviation of each group (n = 5 to 6) and Analysis by Dunnett's test is given in Table 4. Use a 1% significance level.

The combination of the compound A phosphate and the rosiglitazone maleate or pioglitazone hydrochloride in Table 4 significantly reduces the concentration of blood glucose and hemoglobin, which is stronger than the compound A phosphate, rosiglitazone maleic acid. Salt, pioglitazone hydrochloride, MK-0431 are administered alone, and their strength is greater than that of MK-0431 in combination with rosiglitazone maleate or pioglitazone hydrochloride.

Claims (9)

A pharmaceutical composition for treating type 2 diabetes in a mammal comprising humans, the pharmaceutical composition comprising a therapeutically effective amount of rosiglitazone or pioglitazone or a pharmaceutically acceptable salt thereof and (R)-7-[3- Amino-4-(2,4,5-trifluoro-phenyl)-butenyl]-3-trifluoromethyl-5,6,7,8-tetrahydro-imidazo[1,5-a]pyridyl Methyl azine-1-carboxylate or a pharmaceutically acceptable salt thereof, wherein rosiglitazone or a pharmaceutically acceptable salt thereof and (R)-7-[3-amino-4-(2,4, 5-trifluoro-phenyl)-butenyl]-3-trifluoromethyl-5,6,7,8-tetrahydro-imidazo[1,5-a]pyrazine-1-carboxylic acid methyl ester or its The weight ratio of the pharmaceutically acceptable salt is 1:6.25 to 1:75, pioglitazone or a pharmaceutically acceptable salt thereof and (R)-7-[3-amino-4-(2,4,5-three Fluoro-phenyl)-butenyl]-3-trifluoromethyl-5,6,7,8-tetrahydro-imidazo[1,5-a]pyrazine-1-carboxylic acid methyl ester or pharmaceutically acceptable thereof The weight ratio of the accepted salt is from 1:3 to 1:50. The pharmaceutical composition according to claim 1, wherein the (R)-7-[3-amino-4-(2,4,5-trifluoro-phenyl)-butanyl]-3-tri The fluoromethyl-5,6,7,8-tetrahydro-imidazo[1,5-a]pyrazine-1-carboxylic acid methyl ester or a pharmaceutically acceptable salt thereof is used in an amount of 25 per person per day. 300mg. The pharmaceutical composition according to claim 1, wherein the rosiglitazone or a pharmaceutically acceptable salt thereof is used in an amount of from 1 to 12 mg per person per day, and the pioglitazone or a pharmaceutically acceptable salt thereof The daily dose per person is 7.5 to 60 mg. The pharmaceutical composition according to any one of claims 1 to 3, wherein the pharmaceutical composition is a tablet, a hard capsule, a soft capsule, an oral solution, a sustained release agent, a pill, a granule, Granules, sustained release micro Pills or other oral dosage forms. The pharmaceutical composition according to any one of claims 1 to 3, wherein the (R)-7-[3-amino-4-(2,4,5-trifluoro-phenyl)- Methyl butyl hydrazide]-3-trifluoromethyl-5,6,7,8-tetrahydro-imidazo[1,5-a]pyrazine-1-carboxylate or a pharmaceutically acceptable salt thereof is immediate release Or release slowly. The pharmaceutical composition according to any one of claims 1 to 3, wherein the rosiglitazone or pioglitazone or a pharmaceutically acceptable salt thereof is immediate release or slow release. The pharmaceutical composition according to any one of claims 1 to 3, wherein the pharmaceutical composition is a pharmaceutical composition which is administered once a day, twice a day or three times a day. The pharmaceutical composition according to any one of claims 1 to 3, wherein the pharmaceutically acceptable salt is selected from the group consisting of a phosphate, a hydrochloride, a sulfate, a nitrate, a hydrobromide, and a methanesulfonate. Acid salt, maleate, tartrate, succinate, acetate, trifluoroacetate, fumarate, citrate, citrate, besylate, benzoate, naphthalenesulfonic acid Salt, lactate or malate. Use of a pharmaceutical composition according to any one of claims 1 to 8 for the preparation of a medicament for the treatment of diabetes in a mammal including a human.