WO2011009360A1

WO2011009360A1 - Pharmaceutical composition for treatment of type 2 diabetes in mammals including human beings

Info

Publication number: WO2011009360A1
Application number: PCT/CN2010/074309
Authority: WO
Inventors: 袁开红; 马淑芹; 刘华文; 朱亚飞; 李雷明
Original assignee: 江苏恒瑞医药股份有限公司
Priority date: 2009-07-21
Filing date: 2010-06-23
Publication date: 2011-01-27
Also published as: CN101961336A; CN101961336B; HK1153143A1

Abstract

The present invention provides a pharmaceutical composition for treating type 2 diabetes in mammals including human beings, especially a pharmaceutical composition comprising fixed-dose (R)-7-[3-amino-4-(2,4,5-trifluoro-phenyl) -butyryl]-3-trifluoromethyl-5,6,7,8-tetrahydro-imidazo[1,5-a]pyrazin-1-methyl carboxylate or its pharmaceutically acceptable salts and rosiglitazone or pioglitazone or their pharmaceutically acceptable salts, preparation method thereof and use of the pharmaceutical composition for treating 2 type diabetes in mammals including human beings.

Description

Pharmaceutical composition for treating mammals including human type 2 diabetes

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

Human type 2 diabetes is a chronic and progressive disease caused by the complex pathophysiology of double endocrine defects involving insulin resistance and weakened insulin secretion. Treatment for type 2 diabetes generally begins with diet and exercise followed by oral anti-diabetic monotherapy. 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 represent a new class of 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 levels of HbAIC. For a review of the use of DDP-IV for 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) (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 (US5462928, US5543396, W09515309, WO2003004498, WO2003082817, WO2004032836, WO2004085661), wherein the DPP-IV inhibitor MK-0431 produced by Merck has a 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 an 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 an insulin sensitizer, the daily dose of rosiglitazone 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, particularly preferably pioglitazone hydrochloride.

Pharmaceutical dosage forms of a combination of an anti-diabetic agent and a thiazolidinedione derivative have been proposed in the art. For example, EP 0 749 751 teaches a pharmaceutical composition comprising an insulin sensitivity enhancer which is a thiazolyldione compound in combination with other anti-diabetic agents. More particularly, EP 00749751 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 are provided 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 antidiabetic agents and thiazolidinedione derivatives can be found in U.S. Patent No. 6,524. 621; 6, 475, 521; 6, 451, 342 and 6, 153, 632 and PCT patent application WO 0 1/3 594, which are incorporated herein by reference.

Having the following structural formula (R)-7-[3-amino-4-(2,4,5-trifluoro-phenyl)-butanoyl]-3-trifluoromethyl-5,6,7,8-tetra The hydrogen-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.

Formula (I) Summary of the invention

The present invention provides a pharmaceutical composition for treating a mammal, including human type 2 diabetes, 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 1:6.25-1:75, pioglitazone or The weight ratio of the pharmaceutically acceptable salt to the compound of 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, or 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. A specific solid dosage form relates to a tablet containing a combination of a fixed amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof and rosiglitazone or pioglitazone or a pharmaceutically acceptable salt thereof.

The invention also provides a method for preparing a compound of the formula (I) by a drying or a wet treatment method or A method of combining a salt with a fixed dose of a pharmaceutical composition of rosiglitazone or pioglitazone or a salt thereof. 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 a mammal, including human type 2 diabetes, comprising administering to a 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 a diluent; one or more surfactants or wetting agents; one or more disintegrants; and one or more antioxidants.

Salts of 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, milk Acid or malate.

The daily dose of the compound of the formula (I) or a salt thereof incorporated in the pharmaceutical composition of the present invention is 25 mg to 300 mg. Preferably, the daily dose of the compound of formula (I) or a salt thereof is from 50 mg to 200 mg. Discrete human daily doses 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 1 mg to 12 mg, and the daily dose for discrete humans is 1, 2, 4, 8, 10 and 12 mg, pioglitazone or The daily dose of the salt is 7.5 mg ~ 60 mg, and the daily dose of the discrete person is 7.5, 15, 30, 45 and 60 mg. 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, a 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, etc.

The two active pharmaceutical ingredients in the pharmaceutical compositions of the invention may have four release forms:

Ingredients: Compound of formula (I) or its rosiglitazone or pioglitazone

; t or their salts

Immediate release immediate release

Immediate release

Slow release

The pharmaceutical composition of the present invention is prepared by a wet or dry treatment 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 providing tablets with higher radial strength.

The pharmaceutical composition obtained by dry or wet treatment 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 silica, calcium phosphate, magnesium silicate, and talc.

The pharmaceutical compositions of the invention optionally contain one or more binders. Embodiments of the binder include hydroxypropylcellulose (HPC), hydroxypropylmethylcellulose (HMPC), hydroxyethylcellulose, starch 1500, polyvinylpyrrolidone (polypyrrolidone), and copolymerized olefins. Pyrone. A preferred binder is polyvinylpyrrolidone.

The pharmaceutical compositions of the 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 also optionally contain a disintegrant. The disintegrant may be one of several modified starches, modified cellulose polymers or polycarboxylic acids, such as crosslinked hydroxymethylcellulose sodium, sodium starch glycolate, polakolin potassium and hydroxymethylcellulose. CMC Calcium o In one embodiment, the disintegrant is cross-linked hydroxymethylcellulose sodium. Crosslinked hydroxymethylcellulose sodium NF type A is commercially available under the trade name "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 sulfonate, oleyl sulfate, and sodium laurate mixed with stearic acid and talc. Cationic surfactants include benzalkonium chloride and decyltrimethylammonium bromide. Neutral surfactants include glycerol monooleate, polyoxyethylene sorbitan fatty acid esters, polyvinyl alcohol and sorbitan esters. Embodiments of the wetting agent include poloxamers, polyoxyethylene methyl 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, ascorbyl palmitate, citric acid Propyl ester, octyl phthalate, dodecyl phthalate, butylated hydroxytoluene (oxime) and butylated hydroxyanisole (oxime). In one embodiment, the antioxidant is ruthenium or osmium.

A preferred dosage form of the pharmaceutical composition of the invention is a tablet prepared by a compression process. 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 to Opadry® for the formulation of powder blends.

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 released immediately or slowly. freed.

The pharmaceutical tablet composition of the present invention may further contain one or more additional formulation ingredients selected from excipients known in the art of various pharmaceutical preparations. 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 pharmaceutical dosage formulations of all shapes and sizes, whether coated or not. Materials which can be used for coating include hydroxypropylcellulose, hydroxypropylmethylcellulose, titanium dioxide, talc, sweeteners, colorants and flavoring agents.

The invention further provides a method of treating a mammal, including human type 2 diabetes, a therapeutically effective amount of a fixed dose combination pharmaceutical composition of the invention by oral administration of a subject in need of such treatment. 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 (I) or its salt inhibits DPP-IV activity longer than MK-0431, and the inhibition intensity is greater than MK-0431. Through experimental research, the present inventors have surprisingly found that the compound efficacy 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 either component alone, especially when Rogge column The ratio of the ketone or a salt thereof and the compound of the formula (I) or a salt thereof is 1:6.25 to 1:75, and the 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. Therefore, 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 a compound A, code SP2086) or a salt thereof and a composition of pioglitazone or a salt thereof It is of great clinical significance. detailed description

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 Each prescription contains:

Compound A Phosphate 50 mg

Rosiglitazone maleate 4 mg

Microcrystalline cellulose 12 mg

2% starch paddle

Magnesium stearate 0.5 mg Preparation method: Compound A phosphate, rosiglitazone maleate, microcrystalline cellulose was mixed and granulated by a 2% starch slurry. Dry, add magnesium stearate, and mix and compress. Example 2. Compound A phosphate and pioglitazone hydrochloride compound tablet

Each piece contains:

Compound A phosphoric acid

Pioglitazone hydrochloride

Microcrystalline cellulose

2% starch paddle

Magnesium stearate

Preparation method: Compound A phosphate, pioglitazone hydrochloride, microcrystalline cellulose were mixed and granulated by a 2% starch slurry. Dry, add magnesium stearate, and mix and compress. Example 3, Compound A hydrochloride and rosiglitazone maleate compound tablet

Prescription Each tablet contains:

Compound A Hydrochloride 50 mg

Rosiglitazone maleate 15 mg

Microcrystalline cellulose 12 mg

2% starch paddle

Magnesium stearate 0.5 mg Preparation method: Compound A hydrochloride, rosiglitazone maleate, microcrystalline cellulose was mixed and granulated by a 2% starch paddle wet method. Dry, add magnesium stearate, and mix and compress. Example 4: Compound A hydrochloride and pioglitazone hydrochloride compound tablet

Each prescription contains: Compound A Hydrochloride 50 mg

Pioglitazone 15 mg

Microcrystalline cellulose 12 mg

2% starch paddle

Magnesium stearate 0.5 mg Preparation method: Compound A hydrochloride, pioglitazone hydrochloride, microcrystalline cellulose was mixed and granulated by a 2% starch slurry. Dry, add magnesium stearate, and mix and compress. Example 5: Compound A mesylate salt and rosiglitazone maleate compound tablet

Each piece contains:

Compound A methanesulfonic acid

Rosiglitazone maleate

Microcrystalline cellulose

2% starch paddle

Magnesium stearate

Preparation method: Compound A mesylate, rosiglitazone maleate, microcrystalline cellulose were mixed and granulated by a 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 paddle

Magnesium stearate 0.5 mg Preparation method: Compound A methanesulfonate, pioglitazone hydrochloride, microcrystalline cellulose was mixed and granulated by a 2% starch paddle wet method. Dry, add magnesium stearate, and mix and compress. Example 7. Compound A Tartrate and Rosiglitazone Maleate Compound Tablets Each tablet contains:

Compound A Tartrate 50 mg Rosiglitazone maleate 15 mg

Microcrystalline cellulose 12 mg

2% starch paddle

Magnesium stearate 0.5 mg Preparation method: Compound A tartrate, rosiglitazone maleate, microcrystalline cellulose were mixed and granulated by a 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 paddle

Magnesium stearate 0.5 mg Preparation method: Compound A tartrate, pioglitazone hydrochloride, microcrystalline cellulose was mixed and granulated by a 2% starch slurry. Dry, add magnesium stearate, and mix and compress. Example 9. Compound A malate and rosiglitazone maleate compound tablet

Each piece contains:

Compound A malic acid

Rosiglitazone maleate

Microcrystalline cellulose

2% starch paddle

Magnesium stearate

Preparation method: Compound A malate, rosiglitazone maleate, microcrystalline cellulose were mixed and granulated by a 2% starch slurry. Dry, add magnesium stearate, and mix and compress. Example 10 Compound A Malate and Pioglitazone Hydrochloride Compound Tablets Each tablet contains:

Compound A Malate 50 mg Pioglitazone 15 mg Microcrystalline cellulose

2% starch paddle

Magnesium stearate

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

Similarly, a compound containing two active ingredients in different ratios, such as rosiglitazone or a salt thereof and Compound A or a salt thereof, is 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 Methods:

Thaw DPP-IV-Glo. Buffer before use and equilibrate to room temperature. Pre-buffer frozen fluorescein detection reagent, suspend DPP-IV-Glo. Add ultrapure water to the substrate and mix gently to make 1 mM. Substrate, put the fluorescein detection reagent into the brown bottle, add DPP-IV - Glo. The fluorescein detection reagent should be dissolved in 1 minute, dissolve the test compound in DMSO to 50 times the final operating concentration, each test tube Add 50 L of the tested compound 2 L, add 2 L DMSO to the reverse control and the blank control, add 46 L Tris buffer to each tube, and add 48 Tris buffer to the blank control. Add 2 LDPP-IV enzyme to each tube, mix by shaking and centrifuge the tube. Transfer all the material in the test tube to a 96-well plate and mix the substrate with DPP-IV-Glo. at a ratio of 1:49. Mix the vibrations to mix thoroughly. Allow to stand at room temperature for 30-60 minutes before use, add a mixture of 50 DPP-IV-Glo. and substrate in 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 at room temperature for 30 minutes to 3 hours, and measure the chemiluminescence count on a NOVOstar multi-function microplate reader.

[Table 1 ] DPP-IV DPP8 DPP9 Test selection ratio compound Κ ₅₀ (μΜ) Κ ₅₀ (μΜ) (DPP8/DPP-IV) Κ ₅₀ (μΜ) (DPP9/DPP-IV) Compound A 0.008 26.1 3263 75.5 9438

MK-0431 0.019 25.8 1358 92.7 74879 Result: The inhibitory activity of ruthenium on DPP-IV was superior to 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 Ratios of Rosiglitazone and Compound A Pharmaceutical Compositions in Normal ICR Mice Male ICR mice were orally administered with double distilled water and different ratios of rosiglitazone after fasting for 6 hours. Acid salt: 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 for 30 minutes. 2.5g/kg, for oral glucose tolerance test. Blood was taken at 0, 30, 60, and 120 minutes after the sugar was given, and serum glucose levels were measured.

Serum glucose determination method;

Glucose kit was used to determine the glucose content in the serum. 250μ1 enzyme working solution was added, 5μ1 serum was added, and a blank tube (add 5μ1 double distilled water) and standard tube (add 5μ1 glucose standard solution), mix and mix, 37°C water bath After 20 minutes, the blank tube was adjusted to zero, and the colorimetric measurement was performed at OD505nm.

Serum glucose content BG (nmol/1) = OD sample tube / OD standard tube χ5·55

Data processing and statistical analysis;

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

2. Calculate the percentage of blood glucose drop and the area under the curve 30 minutes after the sugar is given.

[Table 2]: Effect of oral test drug composition (10 mg/kg) on oral glucose tolerance in normal ICR mice (mean ± SD, n = 6) Group weight before administration of sugar (nmol/1)

g 0 30 60 120 blank

Control 20.8±1.6 4.61±1.03 12.95±1.89 8.24±1.11 7.11±1.07

1:6.25 20.7±2.0 4.67±1.33 7.06±2.67** 6.10±1.61* 5.13±1.78*

1:12.5 21.0±0.9 4.85±1.02 11.05±2.22 8.41±1.42 7.89±1.07

1:25 22.0±1.1 4.82±1.13 10.22±1.16* 7.93±0.38 7.16±1.17

1:50 21.3±2.2 4.97±0.70 7.33±2.22** 6.02±0.76*** 5.52±0.66*

1:75 21.3±2.3 4.72±1.83 11.81±1.87 8.67±0.97 7.26±0.76

*, PO.05; compared with the blank control group;

**, P<0.01; compared with the blank control group;

***, PO.001; compared with the blank control group; Test Example 3: Oral glucose tolerance study of different ratios of pioglitazone and Compound A pharmaceutical composition to normal ICR mice Male ICR mice were orally administered after 6 hours of fasting Steamed water, different ratios (pioglitazone hydrochloride: compound A phosphate = 7.5:25, 7.5:50, 7.5:75, 7.5:100, 7.5:150, 7.5:375) mixture of the same dose (30mg/kg) Each group was given oral glucose 2.5 g/kg at 30 minutes of administration for oral glucose tolerance test. Blood was taken at 0, 30, 60, and 120 minutes after the sugar was given, and serum glucose levels were measured.

Serum glucose determination method;

Glucose kit was used to determine the glucose content in the serum. 250W enzyme working solution was added, 5μΐ serum was added, and a blank tube (5W double distilled water) and a standard tube (5μΐ glucose standard solution) were added, and mixed, 37°C water bath. After 20 minutes, the blank tube was adjusted to zero, and the colorimetric measurement was performed at OD505nm.

Serum glucose content BG (nmol/1) = OD sample tube / OD standard tube X 5.55 Data processing and statistical analysis;

1. Statistical analysis of data using mean SD and Student-ttest

[Table 3]: Effect of oral test compound (30 mg/kg) on oral glucose tolerance in normal ICR mice (mean SD, n=6) Group weight before administration of sugar (nmol/1)

g 0 30 60 120 blank

Control 26.5±0.8 5.79±0.68 11.99±0.18 9.72±0.78 6.84±0.68

7.5:25 27.0±1.1 6.05±1.04 8.60±1.32*** 8.37±1.24* 6.96±1.90

7.5:50 27.0±0.9 5.82±0.84 7.53±1.21*** 8.41±1.14* 6.01±1.04

7.5:75 26.3±1.5 6.33±0.78 10.34±1.43* 7.08±0.79

7.5:100 27.0±1.5 5.49±0.60 6.94±0.90*** 7.91±0.82** 5.83±1.33

7.5:150 27.2±1.2 5.76±0.69 9.90±1.82* 8.37±0.40** 7.03±1.25

7.5:375 26.5±0.8 6.20±0.9 10.60±2.42 10.40±0.88 6.46±0.65

*, ΡΟ.05; compared with the blank control group;

**, Ρ<0.01; compared with the blank control group;

***, PO.001; compared with the blank control group; Test Example 4: The compound bismuth phosphate, MK-0431 was administered in combination with rosiglitazone maleate or pioglitazone hydrochloride in hereditary obesity and Effect in Diabetic Wistar Rats 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-043K MK-0431 + rosiglitazone maleate, MK-0431 + pioglitazone hydrochloride, compound A phosphate + rosiglitazone maleate and compound A phosphate + pioglitazone salt Acid salt for 14 days. Blood was taken from the tail vein, and blood glucose and hemoglobin A1 were measured by enzymatic method using a commercial kit (NC-ROPET, Nippon Chemiphar CO.). The results were expressed as the mean of each group (n=5-6). The standard deviation is analyzed by Dunnett's test and is given in Table 4. Use a 1% level of significance.

[Table 4]

MK-0431 10mg/kg 291±34 6.0±0.4

MK-0431+Rosiglitazone Maleate 1 Omg/kg+2.5mg/kg 186±14* 4.6±0.5*

MK-0431+Pioglitazone Hydrochloride 10mg/kg+5 mg/kg 162±8* 4.7±0.4* Compound Aphosphate + Rosiglitazone Malay

Acid salt 1 Omg/kg+2.5mg/kg 135±22* 4.3±0.5* Compound A phosphate + pioglitazone hydrochloride

Salt 10mg/kg+5 mg/kg 118±18* 4.1±0.2*

*: P < 0.01 compared with the control group

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

Claims:

A pharmaceutical composition for treating a mammal, including human type 2 diabetes, 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)-butanoyl]-3-trifluoromethyl-5,6,7,8-tetrahydro-imidazo[1,5-a] Methyl pyrazine-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)-butyryl]-3-trifluoromethyl-5,6,7,8-tetrahydro-imidazo[1,5-a]pyrazine-1-carboxylic acid methyl ester or The weight ratio of the pharmaceutically acceptable salt thereof is 1:6.25-1:75, pioglitazone or a pharmaceutically acceptable salt thereof and (R)-7-[3-amino-4-(2,4,5-three Fluoro-phenyl)-butyryl]-3-trifluoromethyl-5,6,7,8-tetrahydro-imidazo[1,5-a]pyrazine-1-carboxylic acid methyl ester or its pharmaceutically The acceptable salt weight ratio is 1:3-1:50.

2. The pharmaceutical composition according to claim 1, wherein the (R)-7-[3-amino-4-(2,4,5-trifluoro-phenyl)-butanoyl]-3-III The daily dose of fluoromethyl-5,6,7,8-tetrahydro-imidazo[1,5-a]pyrazine-1-carboxylic acid methyl ester or a pharmaceutically acceptable salt thereof is 25-300 mg .

The pharmaceutical composition according to claim 1, wherein the rosiglitazone or a pharmaceutically acceptable salt thereof is used in a daily dose of 1 to 12 mg, and the pioglitazone or a pharmaceutically acceptable salt thereof The daily dose is 7.5-60 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, a granule Agent, sustained release pellets or other oral dosage forms.

The pharmaceutical composition according to any one of claims 1 to 4, wherein the (R)-7-[3-amino-4-(2,4,5-trifluoro-phenyl)-butyl group Methyl]-3-trifluoromethyl-5,6,7,8-tetrahydro-imidazo[1,5-a]pyrazine-1-carboxylic acid methyl ester or a pharmaceutically acceptable salt thereof is immediate release Or release slowly.

The pharmaceutical composition according to any one of claims 1 to 5, wherein the roger column Ketones or pioglitazone or their pharmaceutically acceptable salts are either immediate or slow release.

The pharmaceutical composition according to any one of claims 1 to 6, 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 7, wherein the pharmaceutically acceptable salt is selected from the group consisting of phosphates, hydrochlorides, sulfates, nitrates, hydrobromides, methanesulfonic acids Salt, maleate, tartrate, succinate, acetate, trifluoroacetate, fumarate, citrate, citrate, besylate, benzoate, naphthalene Acid salt, lactate or malate.

Use of a pharmaceutical composition according to any one of claims 1-8 for the manufacture of a medicament for the treatment of breast cancer in humans.