TW201226411A - Pharmaceutical composition for the treatment of type 2 diabetes in mammals including human beings - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for the treatment of type 2 diabetes in mammals including human beings, wherein the pharmaceutical composition contains (R)-7-[3-amino-4-(2, 4, 5-trifluo-phenyl)-butyryl]-3-trifluomethyl-5, 6, 7, 8-tetrahydro-imidazo[1, 5-a]pyrazine-1-carboxylic acid methyl ester or its pharmaceutically acceptable salts and rosiglitazone or pioglitazone or their pharmaceutically acceptable salts, preparation method thereof and a method of treating type 2 diabetes in mammals including human beings with the composition.

Description

201226411 VI. Description of the Invention: [Technical Field] The present invention relates to a pharmaceutical composition for treating type 2 sugars in mammals including humans, _ containing rogray = a scientifically acceptable salt and (8)·Η3· Amino _4-(2,4,5_trit-t-l-yl)-3-trifluoromethyl-5,6,7,8-tetrahydro- σm-salt [...] indigenous) strontium butyrate Or the method of (4) attaching to (4) (4) fixing the combination of the county and = preparing the pharmaceutical composition and the use of the pharmaceutical composition for treating type 2 diabetes in a mammal. ·, [Classic] Human type 2 diabetes is a chronic progressive disease caused by complex pathophysiology of double f endocrine deficiency (IV) involving insulin resistance and weakening secretion. The treatment of human type 2 diabetes begins with diet and forging, followed by oral monotherapy for antidiabetic drugs. For many skewers, sweat, this regimen does not adequately control blood glucose during long-term treatment, which leads to the need for combination therapy within a few years after diagnosis. However, the 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 treatment regimen per patient. "Dipeptidyl peptidase_IV (DPP-IV) inhibitors are new agents developed to improve the control of glycemic production in patients with type 2 diabetes for the treatment of patients. In the current clinical trials for the treatment of humans 2 Type 2 diabetes drugs include MK-0431, vildatin (laF-237), saxagliptin (BMS-47718), 3 95093 201226411 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 shown to reduce fasting glucose and postprandial glucose associated with significantly reduced HbAIC levels. Deviations. 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) 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. Pa Tants, 15: 1387-1407 (2005). Currently some DPP-IV inhibitors have been disclosed (US5462928, US5543396, WO9515309, W02003004498, W02003082817, W02004032836, W02004085661), of which the DPP-IV inhibitor MK-0431 produced by Merck It shows good DPP-IV inhibitory activity and selectivity and was launched 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 dose per sputum is usually 951 95093 4 201226411 to 1000 mg, preferably o.l to 500 mg. This dose is administered one 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 the insulin sensitizer, the dose of rosiglitazone per day is usually from 1 to 12 mg, preferably from 2 to 12 mg, and gliacridone is preferably pioglitazone. Pioglitazone hydrochloride or rosiglitazone, pioglitazone maleate or rosiglitazone, particularly preferably pioglitazone hydrochloride. ° Pharmaceutical dosage forms of anti-diabetic agents and combinations of sighs and di-gly derivatives have been proposed in the art. For example, Ep 749751 teaches a pharmaceutical composition comprising an insulin sensitivity enhancing agent which is a bismuth stilbene dione compound in combination with other antidiabetic agents. More particularly, Ep 749 751 teaches that a preferred tensin sensitivity enhancer is, in combination with other antidiabetic agents such as rosiglitazone, phenformin or butyl double pulse, and these The drug is intended to bind (mix and/or coat) conventional excipients to provide taste masking or sustained release. A further example of a combination of an antidiabetic agent and a thiazolidinedione derivative, U.S. Patent No. 6, 'gu, _, which teaches a sustained release form, such as an osmotic pump or a skin patch containing pioglitazone or a koji column _ And a single pharmaceutical composition of rosiglitazone or sputum grid steel. 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, which is incorporated herein by reference. Has the following structural formula (R)|[3_amino group_4_(2,4,5_trisyl_phenyl)_butyl 95093 5 201226411 fluorenyl]-3-trifluoromethyl_5,6,7, 8_tetrahydro-imidazole „ , t θ , TNAX [Ha] 0 is a compound of formula (I) which is a compound of the formula (I), which is disclosed in 200710302335.9.

F

/ _3 Ο Formula (1) [Summary of the Invention] The present invention provides a pharmaceutical composition comprising a human diarrhea, a medicinal composition comprising glitazone or pioglitazone or their pharmacy; 〆, 里,,,, or The pharmaceutically acceptable ratio of 醆盥^ to 式4 /2,|in the rosiglitazone or the pharmaceutically acceptable compound of the formula (1) & or a pharmaceutically acceptable salt thereof From 1. to 1:75, the ratio of pioglitazone or its pharmaceutically (1) compound or its pharmaceutically acceptable compound to the weight of formula (1) and illusion is from 1·3 to 1:5〇. The two pharmaceutical compositions of the present invention are immediate release and the pharmaceutical composition may be in the form of a tablet, and may include a hard leg, a preparation, or other oral administration. Agents (including hard capsules and soft capsules), mouth-to-mouth sputum, sputum, smuggling, liquids, slow release, pills,

ThJ granules, sustained release pellets, and the like. Aspects of the invention relate to dosage forms for the medical administration of a compound of formula (1) or a salt of 95093 6 201226411 and rosiglitazone or a combination thereof. The dosage form may be in a powdery or solid form, and includes a tablet, a capsule, a sachet, etc., which comprises a compound of formula (1) or which is pharmaceutically acceptable; Combined tablets. The present invention is directed to a process for preparing a pharmaceutical composition of a combination of the compound of the formula (1) or a salt thereof and a secret or a salt thereof by a dry or wet treatment method. Dry processing methods include dry compression and dry filament, and wet processing methods include wet granulation. A further aspect of the invention provides a pharmaceutical composition of the invention for treating type 2 diabetes in a mammal, including a human, comprising administering a therapeutically effective amount of a pharmaceutical composition of the invention to a subject in need of said /alpha treatment, In a particular embodiment of the invention, the pharmaceutical composition comprises (1) a compound of formula (1) or a pharmaceutically acceptable salt thereof, which is the first active pharmaceutical ingredient; (2) rosigliflozin or η gegliflozin Or their salts, which are the second active pharmaceutical ingredient; and (3) Run (4) or help (4). 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 agents (combined with one or more diluents; - one or more surfaces) An active agent or wetting agent; one or more disintegrants; and one or more antioxidants. A pharmaceutically acceptable salt of a compound of formula (1), rosiglitazone or a salt of a gegliretide including, but not limited to, _Salt, (10) salt, sulphate, nitrate, 95093 7 201226411 Amino acid salt, methyrin, maleate, tartaric acid, acetate, trifluoroacetate, fumarate, transamate The daily dose per person of the compound of the formula (1) or a salt thereof, which is incorporated in the pharmaceutical composition of the present invention, is an acid salt, a benzoate salt, a benzene acid salt, a naphthyl acid salt, a lactate salt or a malate salt. 25 mg to 3 mg. Compounds of formula (1) = the daily dose per person is preferably 5 g mg to mg. Individual doses per person are 25, 50, 75, 1 〇〇, 15 每日2, 25, and 3 mg of the compound of the formula (I) or a salt thereof. The daily dose of rosiglitazone or its salt incorporated into the solid (four) amount of the present invention is 1 mg to 12 mg per person, and the individual daily doses are 2, 4, 8, 1 G, and 12 mg per person. π gram of glitazone or its salt per person per dose of 7.5 mg to 60 mg, individual daily dose of 7 5, 15 30 45 and 60 mg per person. Rosiglitazone or pi-gigridone Or these daily doses of 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 compound of formula (1) or its salt and Rogge The daily dose of ketone or pioglitazone or a salt thereof is as follows: The compound of the formula (I) or a salt thereof (ton) 25,5〇,75,1〇〇,,200,250,300 Rosiglitazone or a salt thereof (mg) 1,2,4,8,12 pioglitazone or a salt thereof (mg) 7.5, 15, 30, 45, 60 may be any therapeutically effective amount of a compound of the formula (1) or a salt thereof A combination with any therapeutically effective amount of rosiglitazone or pioglitazone or a salt thereof, for example 95093 8 201226411 eg 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 composition of the present invention may have Four forms of release: Ingredients: Compounds of formula (I) Rosiglitazone or ° gliglitazone or a salt thereof or a salt thereof, immediate release, immediate release, slow release, slow release, immediate release, slow release, immediate release, slow release, pharmaceutical composition of the invention The preparation is carried out by a wet or dry treatment method. In one embodiment, the pharmaceutical composition is prepared by a wet treatment method. In one class of this embodiment, the pharmaceutical composition is prepared by a wet granulation process. In wet granulation, high shear/cut granulation or fluidized bed granulation can be applied. In one embodiment, the use of fluidized bed granulation has the advantage of providing the tablet with a higher radial strength. The pharmaceutical composition obtained by the dry or wet treatment method can be compressed into tablets, enclosed, or metered into a sachet. The ice medicine composition contains one or more lubricants or glidants. Examples of moist/moon sputum include magnesium stearate, calcium stearate, stearic acid, sodium oleate, & y test mulch, sputum castor oil or mixtures thereof. A preferred lubricant is magnesium stearate f, a sodium sulphate, or a mixture thereof. Examples of the glidant include colloidal cerium oxide, calcium phosphate, magnesium citrate, and talc. 95093 9 201226411 The pharmaceutical composition of the present invention optionally contains a plurality of agents. Embodiments of the binder include hydroxypropyl cellulose (HPC / 5 mercapto cellulose (HMPC), hydroxyethyl cellulose, propyl propyl hydrazine 1500, polyethyl pyrrolidin steel (polypyridone) mouth copolymerization Isopyrone. Preferred vinylpyrrolidone. The medicinal composition of the present invention may also optionally contain a diluent. The examples of the diluent include mannitol and sorbitol dihydrate. Crystalline cellulose and powdered cellulose. The preferred diluent is microcrystalline cellulose. Microcrystalline cellulose can be obtained from several suppliers, including Sc

Avicel PH 101, Avicel PH 102, Avicei PH 103, Avicel PH 105 and Avicel PH 200 manufactured by 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 hydroxymethylcellulose sodium, sodium starch glycolate, polakolin potassium and hydroxydecyl cellulose. Calcium (CMC Calcium). In one embodiment, the disintegrant is cross-linked sodium strontium cellulose. Crosslinked oxonium silicate type A is commercially available under the trade name "Ac_di_soi". 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 undecyl sulfate, sodium oleyl sulfate, and sodium laurate mixed with stearic acid and talc. Cationic surfactants include benzalkonium chloride and alkyldinonium bromide. Neutral surfactants include glycerol monooleate, polyoxyethylene sorbitan fatty acid esters, polyvinyl alcohol and sorbitan 95093 10 201226411 lipids. Embodiments of the wetting agent include poloxamers, polyoxyacetylene scales, polyethylidene 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-fertility, γ-fertility, δ_tocopherol, although the natural age of the extract, L-ascorbic acid and its sodium or (four), anti-sulphur, palm vinegar, tannic acid Propylene vinegar, citric acid decanoic acid, 2 acid 12 vinegar, butylated η (BHT) and butylated carboxylic acid = methyl ether (BHA). In an embodiment, the antioxidant is BHA. A preferred dosage form of the pharmaceutical composition of the present invention is an agent prepared by a method of thiquoting. The tablet may be coated with a mixture of, for example, propylcellulose and propylmethylcellulose, which contains titanium dioxide and/or other toners such as iron oxide, dyes and lakes; A mixture of a dilute alcohol (ρνΑ) and = ethylene glycol (10) G), containing titanium dioxide and/or other coloring = 'such as emulsified iron, dyes and color killings; or any other suitable instant φ ^ recording agent. The coating provides a taste masking and additional stabilization to the final sheet. The final product, if desired, provided for the preparation of the powder mixture may be added with a sweetener and/or a flavoring agent. The clear rosiglitazone or residual ketone or a salt thereof may be either immediate or slow release, and the compound of formula (1) may be released immediately or slowly. The pharmaceutical tablet composition of the present invention may further contain one or more additional preparations selected from the group of agents known in the field of various pharmaceutical preparations as 95093 11 201226411. Depending on the desired properties of the drug combination, any of the two can be selected individually or in combination based on their known materials in the preparation. Such ingredients include materials such as diluents, compression aids, auxiliaries, slip agents, 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. Substances that can be used for coating include propylcellulose, propylmethylcellulose, titanium dioxide. , talc, sweetener, coloring agent, and flavoring agent. The present invention also provides a therapeutically effective amount of a subject in need of such treatment by oral administration - a fixed dose combination pharmaceutical composition of the invention for treating a mammal, including a human 2 Method of Type 2 Diabetes. 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 or a capsule. A pharmaceutical composition comprising a fixed dose combination The drug can be administered once a day (QD), twice awkward (ΒΠ), or three times a week (Ding Liao). The compound of formula (I) or its salt inhibits DPP_IV activity longer than MK-0431, and the inhibitory strength It is larger than MK_〇431. Through experimental research, the inventors have surprisingly found that the compound efficacy of the compound of the formula (1) or a salt thereof and rosiglitazone or pioglitazone or a salt thereof is remarkably higher than that of any The individual use of the ingredients, especially when the ratio of rosiglitazone or a salt thereof and the compound of the formula or its salt is 1:6.25 to 1:75, the s weekly of the pioglitazone or its salt and the compound of the formula (〗) or its salt A ratio of 1:3 to 1 _5 效果 is more effective. Therefore, a compound of the formula (I) or a salt thereof and a composition of rosiglitazone or a salt thereof, a compound of the formula (I) (hereinafter referred to as a compound A, code number) SP2〇86) or its 12 95093 201226411 :; composition with ngigretin, or a salt thereof, has a clinically significant meaning. ~ [Embodiment] The following examples further describe and illustrate within the scope of the present invention. The examples and the examples are given for the purpose of illustration only, and are not intended to be construed as limiting the invention, and the invention may be variously modified without departing from the spirit and scope of the invention.

Example 1, Compound A Hydrochloride and Rosiglitazone Maleate Compound Tablets Each tablet contains: Compound A Phosphate 50 mg Rosiglitazone Maleate 4 mg Microcrystalline Cellulose 12 mg 2% Starch Pulp suitable amount of magnesium stearate 0.5 mg Preparation method: Compound A phosphate, rosiglitazone maleate, microcrystalline cellulose were mixed and wet granulated with 2% temple powder. Dry, add stearic acid, mix and compress. Example 2: Compound a sulphate and pioglitazone hydrochloride tablet formulation Each tablet contains: Compound A phosphate 50 mg Hydrochloric acid β gegliflozin 15 mg Microcrystalline cellulose 12 mg 95093 13 201226411 Appropriate amount 0.5 mg 2 Preparation method of % starch slurry magnesium stearate: 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 Each tablet contains: Compound A Hydrochloride 50 mg Rosiglitazone Maleate 15 mg Microcrystalline Cellulose 12 mg 2°/ 〇 starch slurry amount of stearic acid town 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 tablet formulation Each tablet contains: Compound A hydrochloride 50 mg Hydrochloric acid π gliglitazone 15 mg Microcrystalline cellulose 12 mg 2% starch slurry Stearic acid Town 0.5 mg 14 95093 201226411 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 oxime sulfonate and rosiglitazone maleate compound tablet Formulation Each tablet contains: Compound A sulfonate 50 mg Rosiglitazone maleate 15 mg

Microcrystalline cellulose 12 mg 2% starch slurry amount of magnesium stearate 0.5 mg Preparation method: Mix the compound A sulfonate 'Rosiglitazone maleate' microcrystalline cellulose, using 2% starch slurry wet method Granulation. Dry 'Add magnesium stearate and mix to compress. Example 6. Formulation of Compound A oxime sulfonate and pioglitazone hydrochloride tablets Each tablet contains: Compound A sulfonate 50 mg Pioglitazone 15 mg Microcrystalline cellulose 12 mg 2% starch slurry Magnesium stearate 0.5 mg Preparation method: Compound A methanesulfonate 'hydrochloric acid ° glitazone' microcrystalline cellulose was mixed and wet granulated with 2% starch slurry. Dry 'Add hard fat 15 95093 201226411 Acid town, mix and compress. Example 7, Compound A Tartrate and Rosiglitazone Maleate Compound Tablets Each tablet contains: Compound A Tartrate 50 mg Maglinic acid rosiglitazone 15 mg Microcrystalline cellulose 12 mg 2% starch slurry Appropriate amount of stearic acid 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 Each tablet contains: Compound A Tartrate 50 mg Hydrochloride ° gliglitazone 15 mg Microcrystalline cellulose 12 mg 2% starch slurry amount of 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. ~ 16 95093 201226411 • Example 9, Compound A Malate and Rosiglitazone Maleate Compound Tablets Each tablet contains: Compound A Malate 50 mg Rosiglitazone Maleate 15 mg Microcrystalline Cellulose 12 mg 2% starch slurry amount of magnesium stearate 0.5 mg Preparation method: Compound A malate, rosiglitazone maleate, • microcrystalline cellulose were 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 Each tablet contains: Compound A malate 50 mg Hydrochloric acid σ-grain S with 15 mg microcrystalline cellulose 12 mg 2% starch slurry Stearic acid Magnesium 0J mg Preparation method: Compound A malate, pioglitazone hydrochloride, and micro-cellulose were uniformly granulated by 2% powder. Dry, add magnesium stearate, and mix and compress. Or Beans, by adjusting the compound A or its salt in the compound, rosiglitazone bis-2, to prepare a complex containing two active ingredients in different ratios: linalone or its salt and compound A Or the ratio of its salt is 95093 17 201226411 1:6.25, 1:12.5, 1:25, 1:50, 1:75, etc. Alternatively, by adjusting the amount of Compound A or its salt, pioglitazone or its salt, To prepare a compound containing different ratios of two active ingredients, for example, the ratio of pioglitazone or its salt to compound A or its salt is 7.5:25, 7.5:50, 7.5:75, 7.5:100, 7.5:150, 7.5:375 Etc. Test Example Γ: In vitro activity and selectivity of Compound A, MK-0431 Study method: Thaw DPP-IV-Glo., buffer before use and equilibrate to room temperature, use buffered frozen luciferin detection reagent before use , suspension DPP-IV-Glo. Add ultrapure water to the matrix and mix gently, make 1 mM matrix, put luciferin detection reagent into the brown bottle, add DPP-IV — Glo. The test reagent should be dissolved within 1 minute, and the test compound dissolved in DMSO to 50 times the final concentration. 50 μg of test compound 2 pL was added to each tube, 2 pL DMSO was added to the negative control and blank control, 46 μ!> Tris buffer was added to each tube, and 48 pL Tris buffer was added to the blank. 2 pLDPP-IV enzyme was added to each tube of the negative control and the test sample, and the tube was shaken and centrifuged. The whole tube was transferred to a 96-well plate, and the mixed matrix and DPP-IV-Glo. : 49. Vibrate and mix until fully mixed. Allow to stand at room temperature for 30 to 60 minutes before use, add 50 pL of DPP-IV — Glo. and substrate mixture to each 96-well plate well, seal with sealant While holding the plate, slowly mix the 96-well material with a plate shaker at 300 to 500 rpm/30 s. Incubate for 30 minutes to 3 hours at room temperature, and measure the chemiluminescence 18 95093 201226411 count value on a NOVOstar multi-function microplate reader. [Table 1] DPP-IV DPP8 DPP9 Test selection ratio ratio compound Ι〇5〇(μΜ) Ι〇50(μΜ) (DPP8/DPP-IV) Ι〇5〇(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: A composition for inhibiting the activity of DPP-I.V than the control drugs _ MK-0431, also stronger than selective MK-0431. Test Example 2: Oral Glucose Tolerance Study of Different Ratios of Rosiglitazone and Compound A Pharmaceutical Compositions to Normal ICR Mice Male ICR mice were orally administered with double distilled water and different ratios of rosiglitazone Ma after 6 hours of fasting. 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 given oral glucose 2.5 minutes after 30 minutes of administration. g/kg, 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, taking 250 μl enzyme working solution, adding 5 μΐ serum, and setting blank tube (adding 5 μΐ double distilled water) and standard tube (adding 5 μ1 glucose standard solution) , mix, 37 ° C water bath for 20 minutes, zero with a blank tube, colorimetric determination at OD505nm. Serum glucose content BG (nmol/1) = OD sample tube / OD standard tube χ 5 · 55 19 95093 201226411 Data processing and statistical analysis; 1, using the standard deviation (Mean Shi SD) and Student-t test Statistical analysis 2. Calculate the percentage of blood glucose drop at 30 minutes after feeding sugar and under the curve

Area AUC [Table 2]: Effect of oral administration of a test drug composition (10 mg/kg) on oral glucose tolerance in normal ICR mice (mean ± standard deviation (Mean ± SD), n = 6) After weight administration (nmol/1) g 0 30 60 120 blank 20.8±1.6 4.61±1.03 12.95±1.89 8.24 ±1.11 7.11 ±1.07 Control 1:6.25 20.7 Soil 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 Earth 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 * J P&lt;0.05 ; compared with the blank control group;氺, P &lt;0.01; compared with the blank control group '***, P < 0.001; compared with the blank control group; Test Example 3: different ratios of pioglitazone and Compound A pharmaceutical composition against 20 95093 201226411 often ICR mice Oral Glucose Tolerance Study Male ICR mice were given double distilled water after 6 hours of fasting, with different ratios (Pygge Levotonide 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 (30 mg/kg), each group administered Oral glucose 2.5g/kg at 30 minutes, and the oral glucose tolerance test was performed. 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, 250μ1 enzyme working solution was added, 5μ1 serum was added, blank tube (add 5μ1 double distilled water) and standard tube (add 5μ1 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 X5.55 data processing and statistical analysis; 1, using the mean ± standard deviation and Student-t test for statistical analysis of the data

2. Calculate the percentage of blood glucose drop and the area under the curve 30 minutes after the administration of sugar. AUC 21 95093 201226411 [Table 3]: Effect of oral administration of test compound (30 mg/kg) on oral glucose tolerance in normal ICR mice (mean ± standard deviation, n=6) Group weight before administration of sugar (nmol/1) g 0 30 60 120 blank control 26.5±0.8 5.79±〇.68 11·99±0'18 9.72±0·78 6.84 ±0.68 7.5:25 27.0 soil 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 10.3 Mud 1.43* 8.65 Soil 0.85* 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 Soil 0.8 6.20±0.9 10·60±2.42 10.40±0.88 6.46 Soil 0.65 * 'P&lt;0.05; compared with the blank control group; **, P&lt;0.01; compared with the blank control group; *** 'P&lt;0.001; compared with the blank control group; Test Example 4: Compound A phosphate, MK-0431 and rosiglitazone maleate or pyridine, respectively Grid Effect of combined administration of ketohydrochloride in Wistar rats with hereditary obesity and diabetes. Male Wistar mice aged 14 to 19 weeks were divided into 9 groups of 5 to 6 rats each taking distilled water and Compound A. Acid salt, rosiglitazone maleate, pioglitazone hydrochloride, MK-0431, MK-0431 + rosiglitazone 22 95093 201226411 Maleate, ΜΚ_0431 + pioglitazone hydrochloride, compound A phosphate + Rogge The ketone maleate salt and the compound A sulphate + pi-siglitazone hydrochloride for 14 days. Blood was collected from the tail vein, and plasma glucose and hemoglobin A1 were separately determined by enzymatic method using a commercial kit (NC-ropet, Nippon Chemiphar CO.). The results were expressed as the average value of each group (n=5 to 6.). The quasi-deviation and the analysis by Dunnett's test are given in Table 4. Use 1〇 / significance level. 〇, • [Table 4] Dose of plasma glucose red blood control 397 ± 35 ' ^ 6.0 ± 〇 4 Compound A phosphate 10 mg / kg 227 ± 58 * ----- 5.9 ± 〇 2 rosiglitazone maleate 2.5mg/kg 339±46 .....^ 6.1±〇4 Pioglitazone hydrochloride 5mg/kg 303±28 5·8±〇.ι MK-0431 10mg/kg 291±34 6·0±〇.4 MK-0431+Rosiglitazone Malay~~~^^ Acidate 10mg/kg+2.5mg/kg 186±14* 4.6±〇5% MK-0431+Pioglitazone Hydrochloride 10mg/kg+5 mg/kg 162±8* 4.7i〇4* Compound A Hydrate + Rogglier 10mg/kg+2.5mg/kg 135±22* ---^ 4.3i〇5% Ketone Maleate Compound A Phosphate + Pyridine Glycine 10 mg / kg + 5 mg / kg 118 ± 18 *, 4.1 ± 〇 2 * keto hydrochloride *: P &lt; 0.01 compared with the control group 95 〇 93 23 201226411 Table 4 in the compound A phosphate and Rogge column The combination of ketone maleate or pioglitazone hydrochloride significantly reduces the concentration of blood glucose and hemoglobin, which is greater than the compound A phosphate, rosiglitazone maleate, pioglitazone hydrochloride, MK-0431. Administered alone, and its intensity is greater than MK-0431 and rosiglitazone maleate or „ The glitazone hydrochloride is used in combination with the drug. [Simple description of the diagram] None. ' [Main component symbol description]. fe 〇 95093 24

Claims (1)

201226411 _ VII. Scope of application: Treatment of type 2 diabetes urinary composition in mammals of humans 'The group (4) contains a therapeutically effective amount of 歹 glitazone or their pharmaceutically acceptable _ _ or . Salt and (r&gt;7 -4·(2,4,5-trifluoro-phenyl)-butenyltrifluoromethane and [1,5 - a]pyrazine-1-carboxylate Methyl ester or a pharmaceutically acceptable salt thereof, wherein rosiglitazone or a pharmaceutically acceptable salt thereof and (r = ke-4-(2,4,5-tris-phenyl)-butanyl &gt;3_: 敦^, 丨,, J 贶 ^ base-5,6,7,8·tetraki-d[l,5-a]pyrazine small carboxylic acid methyl ester or its pharmaceutically acceptable salt weight The ratio is 1:6.25 to 1:75, π 比格刺 or its pharmacy =: fluoromethyl h 6 Μ ^ phenyl) · Τ 基 】 士 广 _ _ 5,6,7,8-tetra- The weight ratio of 5♦ to oxazonic acid or its pharmaceutically acceptable salt is 1:3 to 1:5 〇 2. The pharmaceutical composition as described in claim 凊 patent scope , in the pot, the amine-neutral trifluoro-phenyl)-fusino]_3_trisole =, '7,8 tetrahydro-imidazo[i^a]pyrazine_ι_carboxylic acid oxime ester or The pharmaceutically acceptable salt is used in an amount of from 25 to 300 mg per person per day. The pharmaceutical composition according to claim 1, wherein the Rogge or a pharmaceutically acceptable salt thereof is used in an amount of from 1 to 12 per gram of ketone or a pharmaceutically acceptable amount thereof. Each of the accepted salts is 7.5 to 6 〇mg per day. : : : : : : : : : : : : : : : : : : : : : : : : : : : 医药 医药 医药 医药 医药 医药 医药 医药 医药 医药, soft capsules, oral solutions, sustained release agents, pills, granules, granules, sustained release micro 95093 25 201226411 pills or other oral dosage forms. 5. If the application for the scope of the patent is the third item, and the composition of the medicine, the item is fascinated by the medicine~ 巾3_amine-based ice (2,4,5-three a _ 6 such as Shen Yi 1; acceptable The salt is released immediately or slowly. The drug composition described in any of items 1 to 5 of the ^中Γ^4 rosiglitazone or pioglitazone or their pharmaceutically acceptable salt is immediate release or slow The pharmaceutical composition described in any one of items 1 to 6 of the present invention, and the medical composition of the pharmaceutical composition is administered once a day or once or three times. ' 8. I The pharmaceutical composition according to any one of items 1 to 7, wherein the pharmaceutically acceptable salt is selected from the group consisting of phosphate, hydrochloride, sulfur, salt, salt, hydrogen oxalate, Sucrose, maleate, tartar = salt, alkaloid, acid salt, trifluoroacetate, fumarate, citrate, strontium salt, benzene An acid salt, a benzoate salt, a naphthoate salt, a lactate salt or a malate salt. 9. The use of the pharmaceutical composition according to any one of the claims of the present invention is for the preparation of a treatment including human Drugs for diabetes in mammals. 95093 26 201226411 IV. Designation of representative maps. There is no map in this case. (1) The representative representative of the case is: No. (2) Simple description of the symbol of the representative figure: None. If there is a chemical formula, please reveal the chemical formula that best shows the characteristics of the invention: 2 95093