Detailed description of the invention
The present invention is described pharmaceutical composition of the present invention by following embodiment.
Embodiment 1
I.5-(3-hydroxyl-7-propoxyl group naphthalene-2-base)-1,1-dioxo-[1,2,5] thiadiazolidine-3-ketone potassium salt
Step 1
3-hydroxyl-7-propoxyl group naphthalene-2-propyl formate
3.0g (14.7mmol) 3,7-dihydroxy naphthlene-2-formic acid is added in the suspension of 1.6g (29.6mmol) Feldalat NM in DMA (30mL).By mixture in stirring at room temperature 1 hour, then add 5.05g (29.7mmol) propyl iodide, continue stirring 48 hours.By in mixture impouring water, use 2NHCl acidify.Be extracted with ethyl acetate mixture, by (3 ×) water and saline (1 ×) washing organic facies.Use dried over sodium sulfate solution, under reduced pressure except desolventizing.Residual oil thing carries out purification by flash chromatography dichloromethane, elutes title compound, isolates as yellow oil.MS(M-1)：287。
Step 2
3-benzyloxy-7-propoxyl group naphthalene-2-propyl formate
0.98g (5.73mmol) benzyl bromide a-bromotoluene is added in 1.5g (5.21mmol) 3-hydroxyl-7-propoxyl group naphthalene-2-propyl formate and the mixture of 1.08g (7.81mmol) potassium carbonate in DMF (15mL).By mixture in stirring at room temperature 48 hours, then in impouring water.Be extracted with ethyl acetate mixture, wash organic facies with water (3x) and saline (1x).Use dried over sodium sulfate solution, under reduced pressure except desolventizing.Residual oil thing uses dichloromethane to carry out purification by flash chromatography, elutes title compound, is separated into yellow oil.
Step 3
3-benzyloxy-7-propoxyl group naphthalene-2-formic acid
In the solution of 680mg (1.8mmol) 3-benzyloxy-7-propoxyl group naphthalene-2-propyl formate in EtOH (15mL), add 2.0mL1.0NNaOH, then mixture is stirred 3 hours in 60 DEG C.Under reduced pressure except desolventizing, residual solid is soluble in water.Use MTBE wash solution, with 1NHCl acidify aqueous phase.Filtration gained precipitates, and washing with water, drying under reduced pressure, obtain title compound, is white solid, mp125-128 ° of .MS (M-1): 335.
5-(3-hydroxyl-7-propoxyl group naphthalene-2-base)-1,1-dioxo-[1,2,5] thiadiazolidine-3-ketone potassium salt
In accordance with known methods, title compound is obtained, mp250-255 ° by 3-benzyloxy-7-propoxyl group naphthalene-2-formic acid.
1h-NMR (DMSO-d6: δ 7.88 (s, 1H), 7.54 (d, J=9.04Hz, 1H), 7.13 (s, 1H), 7.12 (d, J=2.64Hz, 1H), 6.98 (dd, J=9.04 and 2.64Hz, 1H), 4.19 (s, 2H), 3.98 (t, 2H), 1.75 (m, 2H), 1.00 (t, 3H) .MS (M-1): 335.
II. (S or R)-3-((R or S)-2-(2 '-fluoro-5 '-methoxyl group-[1,1 '-biphenyl]-4-base) chromane-7-base) butanoic acid
Steps A: (E)-3-(2-(the fluoro-5'-methoxyl group of 2'--[1,1'-biphenyl]-4-base) chromane-7-base) but-2-ene acid methyl ester
At the bromo-2-of N stirred under argon 7-(the fluoro-5'-methoxyl group-[1 of 2'-at 120 DEG C, 1'-biphenyl]-4-base) chromane (1.00g, 2.42mmol, 1.0eq), PdOAc (54.3mg, 0.242mmol), TEA (1.01ml, 7.26mmol, 0.726g/mL, 3.0eq), (Z)-but-2-ene acid methyl ester (727mg, 7.26mmol) and the mixture of tri-o-tolyl phosphine (73.6mg, 0.242mmol) in DMF (15ml) 16 hours.This reactant mixture is cooled to room temperature, then under vacuo except desolventizing.With this reaction of water (15mL) cancellation, and extract twice, this mixture with DCM (15mL).Through the organic layer that anhydrous Na SO drying merges, and concentrated to provide crude product under vacuo.Adopt the eluting crude product of PE:EtOAc=50:1 to provide title compound by silica gel chromatography.
1HNMR(400MHz，CDCl3)δ：7.59(d，J＝7.6，2H)，7.51(d，J＝7.6，2H)，7.11-6.95(m，5H)，6.84-6.82(m，1H)，5.14(d，J＝8.4，1H)，3.82(s，3H)，3.74(s，3H)，3.02-2.82(m，2H)，2.55(s，3H)，2.29-2.04(m，2H).
Step B:3-(2-(the fluoro-5'-methoxyl group of 2'--[1,1'-biphenyl]-4-base) chromane-7-base) methyl butyrate
At room temperature (E)-3-(2-(fluoro-5'-methoxyl group-[1 of 2'-under H atmosphere, 1'-biphenyl]-4-base) chromane-7-base) but-2-ene acid methyl ester (2.40g, 5.55mmol) stir 16 hours with the mixture of Rh/AlO (57.0mg, 0.277mmol) in MeOH (20ml).Then, filter this mixture and concentrated filtrate to provide crude product.This crude product is dissolved in DCM (25mL), and washs with water (20mL).Be separated organic layer, dry through anhydrous Na SO, and concentrated to provide the title compound of the form of mixtures of four kinds of diastereomers under vacuo.
Step C:(S or R)-3-((R or S)-2-(the fluoro-5'-methoxyl group of 2'--[1,1'-biphenyl]-4-base) chromane-7-base) methyl butyrate
Use Chiralcel post OJH250x4.6mmI.D. by SFC, 5um mobile phase: isopropyl alcohol (0.05%DEA) in CO from 5% to 40%
Flow velocity: 2.5mL/min wavelength: 220nm) non-enantiomer mixture of separating step B to be to provide following eluting order: 3-(2-(the fluoro-5'-methoxyl group of 2'-[1,1'-biphenyl]-4-base) chromane-7-base) methyl butyrate (isomer A); 3-(2-(the fluoro-5'-methoxyl group of 2'-[1,1'-biphenyl]-4-base) chromane-7-base) methyl butyrate (isomer B); 3-(2-(the fluoro-5'-methoxyl group of 2'-[1,1'-biphenyl]-4-base) chromane-7-base) methyl butyrate (isomer C); With 3 (2-(the fluoro-5'-methoxyl group of 2'--[1,1'-biphenyl]-4-base) chromane-7-base) methyl butyrate (isomer D).
Step D:
At room temperature stir Lithium hydrate (21.3mg, 0.892mmo) with 3-(2-(the fluoro-5'-methoxyl group-[1 of 2'-, 1'-biphenyl]-4-base) chromane-7-base) mixture of methyl butyrate (isomer A) (40.0mg, 0.0890mmol) in water (1.0ml), THF (1.0ml) and MeOH (1.0ml) 2 hours.Then use DCM (10mL) to dilute this reactant mixture, and wash with water (20ml).Through the organic layer that anhydrous Na SO drying merges, and concentrated to provide crude product under vacuo.(on the GILSON281 instrument being equipped with PhenomenexSynergiC18100*21.2mm*4um, use water and acetonitrile as eluent by preparation HPLC.Mobile phase A: water (containing 0.1%TFA, v/v), Mobile phase B: acetonitrile.Gradient: 60-80%B, 0-10min; 100%B, 10.5-12.5min; 5%B, 13-15min) this crude product of purification to be to provide product.
1HNMR(400MHz，CDCl3)δ：7.56(d，J＝7.6，2H)，7.49(d，J＝7.6，2H)，7.06-6.92(m，3H)，6.78-6.74(m，3H)，5.08(d，J＝9.2，1H)，3.81(s，3H)，3.23-3.20(m，1H)，2.97-2.96(m，1H)，2.80-2.79(m，1H)，2.69-2.56(m，2H)，2.21-2.11(m，2H)，1.30(d，J＝7.2，3H).
By the component I for preparing and II, obtain described diabetes combination of oral medication according to following dosage is composite:
(1) 5-(3-hydroxyl-7-propoxyl group naphthalene-2-base)-1,1-dioxo-[1,2,5] thiadiazolidine-3-ketone potassium salt of 15mg
(2) (S or R)-3-((R or S)-2-(2 '-fluoro-5 '-methoxyl group-[1,1 '-biphenyl]-4-base) chromane-7-base) butanoic acid of 30mg
(3) 268mg microcrystalline Cellulose,
(4) 20mg cross-linking sodium carboxymethyl cellulose, and
(5) 5mg magnesium stearate.
The bull C57BLob/ob mice in 11 week age is housed in reverse smooth circular chamber (from 6:00p.m. to 6:00a.m. illumination) with 6/cage, arbitrarily edible Purina Rodent Meal and drinking-water.At the 1st day, get tail blood sample at 8:00am, measure plasma glucose levels.Animal is randomized into matched group and compound group.The meansigma methods of the plasma glucose levels of each group is mated.Then the Orally administered solvent of animal (there is 0.5% carboxymethyl cellulose of 0.2% tween 80) or the compound (30mg/kg) in solvent is given.Every day, to mice administration, amounts to 3 days.At the 4th day, get basal blood specimens.Adopt the concentration of glucose of YSI2700 dual pathways biochemistry analyzer (YellowSpringsInstrumentCo., YellowSprings, OH) analysed for plasma sample, adopt ELISA algoscopy to measure insulin concentration.Test result shows, its concentration of glucose is 80-120mg/dL, its insulin concentration: 4.4 ~ 8.0mmol/L.
Comparative example 1
Containing the component I of 100mg, not containing component I I, all the other conditions are constant, and its concentration of glucose and insulin concentration are all in the scope of embodiment 1.
Comparative example 2
Containing the component I of 50mg, not containing component I I, all the other conditions are constant, and its concentration of glucose is greater than 150mg/dL.
Comparative example 3
Containing the component I I of 100mg, not containing component I, all the other conditions are constant, and its concentration of glucose and insulin concentration are all in the scope of embodiment 1.
Comparative example 4
Containing the component I I of 50mg, not containing component I, all the other conditions are constant, and its concentration of glucose is greater than 150mg/dL.
Above-described embodiment and comparative example explanation, diabetes combination of oral medication of the present invention, it is composite by two kinds of dissimilar active matters, under the prerequisite reaching identical therapeutic effect, the use amount of the active matter of medicine is reduced at least 1/2, when two kinds of active matters that the present invention adopts are used alone, within the scope of equal volume, glucose in blood cannot be reduced in normal range of the present invention, prove to create cooperative effect between two kinds of active matters that the present invention adopts.
Applicant states, the present invention illustrates diabetes combination of oral medication of the present invention by above-described embodiment, but the present invention is not limited to above-mentioned specific components and formula, does not namely mean that the present invention must rely on above-mentioned detailed component and formula could be implemented.Person of ordinary skill in the field should understand, any improvement in the present invention, to equivalence replacement and the interpolation of auxiliary element, the concrete way choice etc. of each raw material of product of the present invention, all drops within protection scope of the present invention and open scope.