it was found surprisingly by the inventors that pinocembrin had two different crystalline forms α and β , and the inventors developed the preparations thereof . the inventors also found that there was a significant difference between the two crystalline forms in the uptake by organism , wherein the uptake rate of β crystalline form was greater than that of α crystalline form , for example , the uptake rate of β crystalline form can be 2 times or more greater than that of α crystalline from . the biological activities thereof in medicament therapy are different due to differences in drug blood concentrations . one embodiment of the present invention provides α crystalline form , β crystalline form , or the mixture of α crystalline form and β crystalline form in different proportions , with respect to pinocembrin . preferably , these crystalline forms or the mixture thereof contains no crystal water or other organic solvents . one embodiment of the present invention provides the method for preparing α crystalline form , β crystalline form , or the mixture of α crystalline form and β crystalline form of pinocembrin in different proportions . one embodiment of the present invention provides a pharmaceutical composition comprising pure α crystalline form , pure β crystalline form , or the mixture of α crystalline form and β crystalline form of pinocembrin in different proportions . the pharmaceutical composition can also comprise one or more pharmaceutically acceptable carriers . there is no limitation on the pharmaceutically acceptable carriers , provided that they are suitable for formulation and would not substantially affect the effect of pinocembrin crystalline form of the present invention . one embodiment of the present invention relates to dosage forms comprising the pinocembrin crystalline forms solid . there are no limitations on specific dosage forms . for example , they can be tablets , capsules , pills , injections , sustained - release preparations , controlled - release preparations and the like . one embodiment of the present invention provides use of the crystalline forms solid of pinocembrin , including α crystalline form , β crystalline form or the mixture of α crystalline form and β crystalline from in different proporations , for bringing about difference of drug uptake in treatment . one embodiment of the present invention relates to the use of α crystalline form , β crystalline form , or the mixture of α crystalline form and β crystalline form of pinocembrin in different proportions in the manufacture of a medicament for treating diseases related to cerebral ischemia or a medicament for preventing diseases related to cerebral ischemia by protecting neurovascular unit function . one embodiment of the present invention provides the use of pinocembrin for protecting neurovascular unit function in the treatment of diseases related to cerebral ischemia , and for improving the blood concentration in an organism due to the crystal form effect . morphologic features of a sample of α crystalline form of pinocembrin according to an embodiment : for a sample of α crystalline form of pinocembrin obtained according to an embodiment of the present invention , when analyzed by x - ray single crystal diffraction , it showed monoclinic crystal system symmetry , the space group was p2 1 / c , and the crystal cell parameter values were as follows : a = 5 . 189 å , b = 24 . 149 å , c = 10 . 472 å , α = 90 °, β = 102 . 31 ° and γ = 90 °. fig1 is an illustration showing the relative configuration of the molecule , fig2 is an illustration showing the stereo - structure projection of the molecule , fig3 is an illustration showing the unit cell stacking of the molecule along the a axis . table 1 shows atomic coordinate parameters and equivalent temperature factors . table 2 shows bond length values of bonding atoms . table 3 shows bond angle values of bonding atoms . as some carbon atoms of b ring adopt a disordered orientation state , the four atoms of c 2 ′ , c 3 ′ , c 5 ′ and c 6 ′ occupy two positions , with an occupancy rate of 0 . 5 respectively . note : in table 2 and 3 , for the atoms c 2 ′ , c 3 ′ , c 5 ′ and c 6 ′ of b ring , only the bond length and bond angle values of one position are given in table 2 and 3 . powder ( polycrystal ) x - ray diffraction ( cuk α radiation ) was performed on the α crystalline form solid of pinocembrin , and the diffraction peak locations : 2 - theta value (°) or d value ( å ), and the relative strength of diffraction peak : peak height value ( height %) or peak area value ( area %) show the following characteristics ( see table 4 , fig4 ). ir analysis with kbr pellet was performed on α crystalline form solid of pinocembrin ( see fig6 ). the characteristic peaks were as follows : 3090 . 6 , 3011 . 6 , 2889 . 1 , 2747 . 4 , 2636 . 2 , 1631 . 5 , 1602 . 5 , 1584 . 3 , 1487 . 7 , 1466 . 2 , 1454 . 5 , 1435 . 6 , 1354 . 9 , 1302 . 4 , 1257 . 0 , 1217 . 0 , 1168 . 2 , 1088 . 6 , 1064 . 9 , 1028 . 0 , 1014 . 6 , 1001 . 3 , 975 . 8 , 918 . 0 , 887 . 7 , 861 . 8 , 825 . 9 , 789 . 9 , 766 . 4 , 715 . 2 , 698 . 1 , 663 . 7 , 646 . 7 , 620 . 3 , 587 . 3 , 574 . 9 , 560 . 5 , 526 . 9 and 487 . 9 cm − 1 , wherein the peaks of 2891 . 1 , 2747 . 4 , 2636 . 2 , 1631 . 5 and 1354 . 9 cm − 1 were the main characteristic peaks of α crystalline form solid of pinocembrin . morphologic features of a sample of β crystalline form of pinocembrin according to an embodiment : for β crystalline form of pinocembrin obtained according to an embodiment of the present invention , when analyzed by powder ( polycrystal ) x - ray diffraction ( cuk α radiation ), it showed the diffraction peak location : 2 - theta value (°) or d value ( å ) and the relative strength of diffraction peak : peak height value ( height %) or peak area value ( area %), shown as the following characteristic peak values ( see table 5 , fig7 ) ir analysis with kbr pellet was performed on β crystalline form solid of pinocembrin according to an embodiment of the present invention ( see fig9 ). the characteristic peaks were as follows : 3090 . 8 , 2890 . 0 , 2748 . 9 , 2638 . 3 , 1633 . 5 , 1602 . 9 , 1585 . 0 , 1487 . 9 , 1466 . 1 , 1454 . 3 , 1344 . 4 , 1302 . 7 , 1216 . 7 , 1168 . 2 , 1088 . 4 , 1065 . 5 , 1028 . 8 , 1014 . 3 , 1001 . 5 , 975 . 8 , 917 . 8 , 888 . 2 , 861 . 8 , 826 . 6 , 789 . 1 , 766 . 6 , 741 . 1 , 715 . 4 , 698 . 0 , 663 . 7 , 646 . 0 , 620 . 5 , 587 . 9 , 574 . 8 , 560 . 9 , 527 . 2 and 488 . 4 cm − 1 , wherein the peaks of 2890 . 0 . 2748 . 9 . 2638 . 3 , 1633 . 5 and 1344 . 4 cm − 1 were the main characteristic peaks of β crystalline form solid of pinocembrin . method for preparing a sample of α crystalline form of pinocembrin according to an embodiment of the present invention ( 1 ) dissolving a sample completely in a solvent selected from the group consisting of methanol , ethanol , chloroform , acetone , ethyl acetate , n - butanol , isopropanol , acetonitrile , thf , dioxane , 95 % ethanol , glacial acetic acid , formic acid , ether , dichloromethane , toluene , benzene , n - hexane , cyclohexane , dioxane , dmf , petroleum ether , ammonia , n - propanol , or a mixture thereof , then , ( a ) placing the mixture in a condition of temperature 4 - 50 ° c . and relative humidity 10 %- 75 % to allow recrystallization for 1 to 60 days , or ( b ) adding water to allow precipitate , then obtaining α crystalline form of pinocembrin through filtration under reduced pressure , freeze - drying or cold spray . method for preparing a sample of β crystalline form of pinocembrin according to an embodiment of the present invention using a sample of α crystal form solid as the material , then ( b ) dissolving the material completely in a solvent of pyridine or dmso , adding water to allow precipitate , and performing filtration under reduced pressure , freeze - drying or cold spray . pure α crystalline form , pure β crystalline form or the mixture of α and β crystalline forms of pinocembrin in any proportion according to the present invention have an effect on treating diseases related to cerebral ischemia or preventing diseases related to cerebral ischemia by protecting neurovascular unit function . there is a difference in bioavailability between pure α crystalline form and pure β crystalline form of present invention . for oral administration , the bioavailability of β crystalline form is more than 2 times higher than that of α crystalline form . for a mixture of the two crystalline forms in any proportion , the bioavailability thereof can vary , dependent on different contents of β crystalline form . for a pharmaceutical composition or a preparation comprising pure α crystalline form , pure β crystalline form or a mixture thereof in any proportion of pinocembrin according to an embodiment of present invention , daily dosage is 5 ˜ 250 mg , based on pinocembrin crystalline forms solid . preparations include tablets , capsules , pills , injections , sustained - release preparations , controlled - release preparations and the like . fig1 is an illustration showing the relative configuration of the molecule . fig2 is an illustration showing the stereo - structure projection of the molecule . fig3 is an illustration showing the unit cell stacking of the molecule ( along the a axis ). fig4 is a powder x - ray diffraction pattern for a sample of α crystalline form of pinocembrin . fig5 is a dsc trace for a sample of α crystalline form of pinocembrin . fig6 is an infrared absorption spectra for a sample of α crystalline form of pinocembrin . fig7 is a powder x - ray diffraction pattern for a sample of β crystalline form of pinocembrin . fig8 is a dsc trace for a sample of β crystalline form of pinocembrin . fig9 is an infrared absorption spectra for a sample of β crystalline form of pinocembrin . the following exemplary examples are provided for the purpose of better description of the present invention , however , it should be appreciated that the present invention is not limited to the given examples . instrument : mac dip - 2030k area detector test condition : tube voltage : 50 kv , tube flow : 80 ma , ω scanning , mok α , 2θ ≦ 50 . 0 °, scan range : 0 - 180 °, pivot angle : 6 °, step : 6 °, scan rate : 1 . 8 °/ min . instrument : rigaku d / max 2550 powder x - ray diffractometer test conditions : voltage : 40 kv , current : 150 ma , scan rate : 8 °/ min . instrument type : seiko instruments inc . differential scanning calorimeter test conditions : purge gas : n 2 , heating rate : 10 ° c . / min . temperature range : 25 - 250 ° c . instrument nicolet ft - ir spectrometer : impact 400 test conditions : kbr pellet instrument : shimadzu lc - 10avp high performance liquid chromatography , spd - m10avp diode array detector , class - vp chromatography data system ; column : alltch c 18 ( 5μ , 150 × 4 . 6 mm ); test conditions : column temperature : room temperature ; wavelength : 290 nm ; mobile phase : methanol / phosphate saline ph 3 . 0 ( 64 / 36 ); flow rate : 1 . 0 ml / min ; injection volume : 20 μl ; injection concentration : 500 . 0 μg / ml to 1000 ml hydrogenation reaction kettle , was added 5 g ( 19 . 7 mmol ) of 5 , 7 - dihydroxyflavone , 650 ml of anhydrous ethanol , and 1 . 5 g of 10 % palladium on carbon . under a hydrogen pressure of 0 . 13 mpa , the reaction was performed for 4 h at 40 ° c . when the reaction was finished , the palladium on carbon was filtered . the filtrate was concentrated , then separated and purified by column chromatography ( eluted by methanol : acetic ether : petroleum ether = 2 : 10 : 100 ( v : v : v )) under vacuum . the solvent was evaporated to dryness , and 3 . 9 g of white amorphous solid powder was obtained ( purity : 98 . 6 %, detected by hplc ) in a yield of 52 % [ 2 ] . method 1 for preparing a sample of α crystalline form of pinocembrin 5 g pinocembrin sample was added to 20 ml of 95 % ethanol , and was heated to be dissolved completely , then cooled to room temperature and allowed to stand for 24 h . white solid was precipitated , and was filtered and dried . 4 . 5 g of white crystalline ( purity : 98 . 8 %, detected by hplc ) was obtained in a recovery of 90 %. the obtained crystalline was analyzed by x - ray single crystal diffraction . it showed monoclinic symmetry , space group was p2 1 / c , and the crystal cell parameter values were a = 5 . 189 å , b = 24 . 149 å , c = 10 . 472 å , α = 90 °, β = 102 . 31 ° and γ = 90 °. powder ( polycrystal ) x - ray diffraction ( cuk α radiation ) was performed on the obtained crystalline . the characteristic peak values of the diffraction peak location : 2 - theta value (°) or d value ( å ) and the relative strength of diffraction peak : peak height value ( height %) or peak area value ( area %) were shown in table 4 , and the obtained trace was shown in fig4 . dsc analysis was performed on the obtained crystalline , and decalescence transition temperature was 206 ° c . ir analysis with kbr pellet was performed on the crystalline obtained , and the characteristic peaks were as follows : 3090 . 6 , 3011 . 6 , 2889 . 1 , 2747 . 4 , 2636 . 2 , 1631 . 5 , 1602 . 5 , 1584 . 3 , 1487 . 7 , 1466 . 2 , 1454 . 5 , 1435 . 6 , 1354 . 9 , 1302 . 4 , 1257 . 0 , 1217 . 0 , 1168 . 2 , 1088 . 6 , 1064 . 9 , 1028 . 0 , 1014 . 6 , 1001 . 3 , 975 . 8 , 918 . 0 , 887 . 7 , 861 . 8 , 825 . 9 , 789 . 9 , 766 . 4 , 715 . 2 , 698 . 1 , 663 . 7 , 646 . 7 , 620 . 3 , 587 . 3 , 574 . 9 , 560 . 5 , 526 . 9 and 487 . 9 cm − 1 . the above spectra data showed that the crystalline form obtained in the present example was α crystalline form . methods 2 to 10 for preparing a sample of α crystalline form of pinocembrin referring to the preparation method of example 1 , using ethyl acetate , chloroform , acetone , acetonitrile , thf , ether , benzene , cyclohexane or dmf as the solvent , white crystalline of pinocembrin was obtained . the results of the experiments were shown in table 6 . powder x - ray diffraction , dsc and ir analysis were performed on the obtained crystalline , and the results showed that the crystalline form was α crystalline form of pinocembrin . 5 g pinocembrin sample was dissovled in 100 ml mixture of 95 % ethanol and acetone ( 95 % ethanol : acetone = 1 : 1 ) completely at room temperature , then 100 ml water was added under stirring , and white precipitate appeared . the precipitate was filtered under reduced pressure and dried to obtain 4 . 00 g white crystalline ( purity : 98 . 7 %) in a recovery of 80 . 0 %. powder x - ray diffraction , dsc and ir analysis were performed on the obtained crystalline form , and the results showed that the crystalline form obtained was α crystalline form of pinocembrin . methods 12 to 16 for preparing a sample of α crystalline form of pinocembrin referring to the preparation method of example 1 , using the mixture of isopropanol and thf ( isopropanol : thf = 2 : 1 ), the mixture of acetonitrile and dmf ( acetonitrile : dmf = 4 : 1 ), the mixture of methanol and acetone ( methanol : acetone = 3 : 2 ), the mixture of ethanol and acetonitrile ( ethanol : acetonitrile = 1 : 1 ), and the mixture of ethanol , acetone and glacial acetic acid ( ethanol : acetone : glacial acetic acid = 2 : 1 : 0 . 1 ) as the solvents , white crystalline of pinocembrin was obtained . the results of the experiments were shown in table 7 . powder x - ray diffraction , dsc and ir analysis were performed on the obtained crystalline , and the results showed that the crystalline form obtained was α crystalline form of pinocembrin . 10 g sample of α crystalline form of pinocembrin was placed in a mortar , grinded evenly in the same direction for 1 hour at room temperature , and white crystalline was obtained , which is different from α crystalline form . powder ( polycrystal ) x - ray diffraction ( cuk α radiation ) was performed on the obtained crystalline . the characteristic peak values of the diffraction peak location : 2 - theta value (°) or d value ( å ) and the relative strength of diffraction peak : peak height value ( height %) or peak area value ( area %), were shown in table 5 , and the obtained trace was shown in fig7 . dsc analysis was performed on the obtained crystalline , and decalescence transition temperature was 204 ° c ., as showed in the dsc trace . ir analysis with kbr pellet was performed on the crystalline obtained , and the characteristic peaks were as follows : 3090 . 8 , 2890 . 0 , 2748 . 9 , 2638 . 3 , 1633 . 5 , 1602 . 9 , 1585 . 0 , 1487 . 9 , 1466 . 1 , 1454 . 3 , 1344 . 4 , 1302 . 7 , 1216 . 7 , 1168 . 2 , 1088 . 4 , 1065 . 5 , 1028 . 8 , 1014 . 3 , 1001 . 5 , 975 . 8 , 917 . 8 , 888 . 2 , 861 . 8 , 826 . 6 , 789 . 1 , 766 . 6 , 741 . 1 , 715 . 4 , 698 . 0 , 663 . 7 , 646 . 0 , 620 . 5 , 587 . 9 , 574 . 8 , 560 . 9 , 527 . 2 and 488 . 4 cm − 1 . the above spectra date showed that the crystalline form obtained in the present example was β crystalline form . at room temperature , 5 g pinocembrin sample was dissolved in 75 ml dmso , then to which 150 ml of water was added under stirring , and white precipitate appeared . the precipitate was filtered and dried to obtain 4 . 2 g white crystalline ( purity : 98 . 8 %, detected by hplc ) in a recovery of 84 . 0 %. powder x - ray diffraction , dsc and ir analysis were performed on the obtained crystalline , and the results showed that the crystalline form obtained was β crystalline form of pinocembrin . method 3 for preparing a sample of β crystalline form of pinocembrin except that pyridine was used as the solvent , the same preparation method as that of example 18 was used , and 8 . 8 g white crystalline was obtained ( purity : 98 . 6 %, detected by hplc ) in a recovery of 88 . 0 %. powder x - ray diffraction , dsc and ir analysis were performed on the obtained crystalline , and the results showed that the crystalline form obtained was β crystalline form of pinocembrin . method for preparing a sample of pinocembrin mixture of α crystalline form and β crystalline form in a ratio 1 : 1 10 g of pinocembrin sample α and β crystalline form were respectively weighed and placed into a sealable vessel . the vessel was sealed and shook to mix the solid evenly . the mixture sample of α and β crystalline form in a ratio 1 : 1 was obtained . the samples of pure α crystalline form , pure β crystalline form , or solid mixture of ( α + β ) crystalline form ( α : β = 1 : 1 ) of pinocembrin were mixed with excipients in different proportions to obtain a solid of the combined pharmaceutical active ingredients . tablets containing 5 ˜ 60 mg of active ingredients were prepared . the tablet formula were given in table 8 . the detailed preparation method was as follows : mixing the excipients with pinocembrin evenly , then adding appropriate amount of 1 % sodium hydroxymethyl cellulose solution to make a dough . the dough was screened to obtain granules . the wet granules were dried and sieved . then magnesium stearate and talc powder were added and mixed evenly , and the product was obtained by tabletting . the samples of pure α crystalline form , pure β crystalline form , or solid mixture of ( α + β ) crystalline form ( α : β = 1 : 1 or 1 : 3 ) of pinocembrin were mixed with excipients in different proportions to obtain a solid of the combined pharmaceutical active ingredients . the capsules containing 5 ˜ 60 mg of active ingredients were prepared . the capsule formulas were given in table 9 . the detailed preparation method was as follows : mixing the excipients with pinocembrin evenly , then appropriate amount of 1 % sodium hydroxymethyl cellulose solution was added to make wet granules , the wet granules were dried and sieved . then magnesium stearate was added and mixed evenly , and the product was obtained by filling the mixture above into empty capsules . alternatively , without granulation , the product was obtained by filling the excipients and pinocembrin into empty capsules directly , after they were mixed evenly and sieved . method 3 for preparing combined pharmaceutical formulation ( injection solution and freeze - dried powder for injection ) pure α crystalline form , pure β crystalline form , or solid mixture of ( α + β ) crystalline form ( α : β = 1 : 1 ) of pinocembrin were mixed with excipients in different proportions to obtain a solid of the combined pharmaceutical active ingredients . then the injections containing 5 ˜ 60 mg of active ingredients per ampoule were obtained . the injection formulas were given in table 10 . ( 3 ) the mixed solution was stirred magnetically for 20 min at 40 ˜ 50 ° c . when the solution became clear and transparent , 0 . 5 g of actived carbon for injection was added . then the mixture was heated to 80 ° c . with stirring and kept at this temperature for 15 min , then the carbon was filtered . the filtrate was subpackaged into ampoule at 4 ml each . pinocembrin injection solution was obtained after sterilizing at 121 ° c . for 15 min . ( 1 ) in the sterile room , 40 g of hydroxypropyl - β - cyclodextrin was weighed and dissolved in water to make 150 ml solution . 0 . 1 g actived carbon was added , and the mixture was heated to mild boiling for 15 min , and then the carbon was filtered out ; ( 2 ) 1 g of pinocembrin was dissolved in 20 ml anhydrous ethanol , then the obtained solution was added into the hydroxypropyl - β - cyclodextrin solution mentioned above ; ( 3 ) the mixture was stirred magnetically for 20 min at 40 ˜ 50 ° c .- when the solution became clear and transparent , the inclusion complex solution of pinocembrin in hydroxypropyl - β - cyclodextrin was obtained ; ( 4 ) water was added to the inclusion complex solution to 200 ml . the mixture was filtered through 0 . 22 μm filter membrane . the filtrate was subpackaged into vial of 10 ml ( 2 ml / vial ), and placed into freeze dryer to freeze - dry . the sterile powder for injection was obtained after the vials &# 39 ; stoppers were sealed . ( 1 ) to 200 ml of distilled water , 20 g of hydroxypropyl - β - cyclodextrin was added and dissolved with stirring . 0 . 5 g actived carbon for infusion was added . the mixture was heated to 80 ° c . with stirring and kept at this temperature for 15 min , then the carbon was filtered ; ( 2 ) 1 g of pinocembrin was weighed and dissolved in 20 ml anhydrous ethanol , then the obtained solution was poured into the hydroxypropyl - β - cyclodextrin solution mentioned above ; ( 3 ) the mixture was stirred magnetically for 20 min at 40 ˜ 50 ° c . when the solution became clear and transparent , the inclusion complex solution of pinocembrin in hydroxypropyl - β - cyclodextrin was obtained ; ( 4 ) water was added to the inclusion complex solution to 800 ml . after adding 90 g sodium chloride for injection , the solution was adjusted to a ph of 8 ˜ 9 and diluted to 10000 ml with water . then 10 g actived carbon for injection was added , and stirred for 20 min ; ( 5 ) after the carbon was removed , the solution was subpackaged in 100 ml per bottle . the product was obtained after sterilizing at 121 ° c . for 30 min . ( 1 ) 20 g of hydroxypropyl - β - cyclodextrin was added to 200 ml of distilled water and dissolved with stirring . 0 . 5 g actived carbon for infusion was added . the mixture was heated to 80 ° c . by stirring and kept at this temperature for 15 min , then the carbon was filtered ; ( 2 ) 1 g of pinocembrin was dissolved in 20 ml anhydrous ethanol , then the obtained solution was poured into the hydroxypropyl - β - cyclodextrin solution mentioned above ; ( 3 ) the mixture was stirred magnetically for 20 min at 40 ˜ 50 ° c . when the solution became clear and transparent , the inclusion complex solution of pinocembrin in hydroxypropyl - β - cyclodextrin was obtained ; ( 4 ) water was added to the inclusion complex solution to 800 ml . after adding 500 g glucose for injection , the solution was adjusted to a ph of 8 ˜ 9 and diluted to 10000 ml with water . then 10 g actived carbon for injection was added , and stirred for 20 min ; ( 5 ) after the carbon was removed , the solution was subpackaged in 100 ml per bottle . the product was obtained after sterilizing at 121 ° c . for 30 min . in vivo absorption and blood concentration characteristics for pinocembrin solid active ingredient of α and β crystalline forms : 18 of sd rats , female and male each half , with body weight of 230 ˜ 250 g , were randomized into 3 groups , with 6 rats each group and female half . after 10 hours of fasting but free water intake , the rats were administrated with pinocembrin solid active ingredient powder of α , β or 1 : 1 ( α + β ) mixed crystalline forms , at a dose of 50 mg / kg to stomach . then the arterial blood samples at different times were taken and the contents of pinocembrin were determined . the result showed that for pinocembrin of different crystalline forms , at the same dosage by oral administration , the blood concentrations and the time to reach the peak concentration were different , wherein the blood concentration of α crystalline form was obviously lower than that of β crystalline form .
two crystalline forms of pinocembrin of formula : α and β , their preparation and their use for manufacture of pharmaceutical compositions . there exists difference between them in bioavailability . they are used for treating and preventing cerebral ischemic diseases by protective action of neurovascular unit , and enhancing blood drug level in vivo .