Abstract:
Disclosed are a complex of glucose derivative and proline, a crystal, a preparation method and a use. In an X-ray powder diffraction diagram of the eutectic crystal when the diffraction angle is 2θ, characteristic diffraction peaks comprise 4.339, 11.499, 12.835, 13.921, 15.294, 16.212, 16.804, 17.154, 18.335, 19.274, 19.982, 22.710, 23.218, 24.885, 27.940, 29.612 and 30.313, and the 2θ error range is ±0.1. The method comprises: mixing a compound A solution with an L-proline solution, and performing cooling and crystallization. The present invention further provides a of the crystal in medicine preparation. The eutectic crystal in the present invention features high water-solubility, low hygroscopicity and high stability, and is suitable for manufacturing a variety of preparations.

Description:
FIELD OF INVENTION 
       [0001]    The present invention relates to a complex of a glucose derivative and proline, eutectic crystal, preparation method and use; in particular to a complex of (2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-((3-fluorooxetan-3-yl)methoxy)benzyl)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol and bis(L-proline), eutectic crystal, preparation method and use. 
       PRIOR ARTS 
       [0002]    Sodium-dependent glucose co-transporters (SGLTs) play a key role in maintaining human plasma glucose stable. SGLTs have been found in intestine (SGLT1) and kidney (SGLT1 and SGLT2). Renal SGLT reabsorbs glucose from renal filtrate, thereby preventing the loss of glucose from urine. 98% of glucose is reabsorbed in the kidney by SGLT2, and only the remaining 2% is reabsorbed by SGLT1. Inhibition of SGLT2 can specifically inhibit the re-absorption of glucose by kidney and increase the excretion of glucose in the urine, which may normalize the plasma glucose for diabetics. Therefore, the inhibitors of SGLT, particularly SGLT2, are promising candidates for anti-diabetic drugs (Handlon, A. L., Expert Opin. Ther. Patents (2005) 15(11):1531-1540). 
         [0003]    So far, a lot of pharmaceutical companies have developed a series of SGLT2 inhibitors, such as those described in: Handlon, A. L., Expert Opin. Ther. Patents (2005) 15(11):1531-1540; William N. W., Journal of Medicinal Chemistry, 2009, Vol. 52, No. 7, 1785-1794; Chao, E. C. et al., Nature Reviews Drug Discovery, 2010, Vol. 9, No. 7, 551-559. Aryl glycosides as SGLT2 inhibitors are also known by the following patent applications: WO 01/27128, WO 02/083066, WO 03/099836, US 2003/0114390, WO 04/063209, WO 2005/012326, US 2005/0209166, US 2006/0122126, WO 2006/011502, US 2007/0293690, WO 2008/034859, WO 2008/122014 and WO 2009/026537. 
         [0004]    A SGLT2 inhibitor (2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-((3-fluorooxetan-3-yl)methoxy)benzyl)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol (hereinafter, referred to as “compound A”) is described in an international patent application WO2012/109996 and has a chemical structure of formula A: 
         [0000]    
       
                 
         
             
             
         
       
     
         [0005]    The compound A prepared by a method described in the patent is amorphous and hygroscopic, which gains 14.72% weight under 95% RH. After absorption of moisture, it is allochroic usually and easy to lose stability and decompose, which is not conducive to the operation of preparation. Therefore, it is necessary to develop an eutectic crystal of the compound A with excellent physical and chemical properties and is conducive to the operation of preparation. 
       CONTENT OF THE PRESENT INVENTION 
       [0006]    The technical problem to be solved by the present invention is to provide a complex of a glucose derivative and proline, crystal, preparation method and use. The eutectic crystal of the glucose derivative (compound A) and proline in the present invention has better physical and chemical properties, better water solubility, lower hygroscopicity, higher stability and is particularly suitable for the preparation of various preparations. The structure of compound A is as follows: 
         [0000]    
       
                 
         
             
             
         
       
     
         [0007]    During the study of preparing the eutectic crystal of compound A, the inventors of the present invention have found that it can only afford jelly, oil or amorphous substance, etc. other than the eutectic crystal of compound A with good physical and chemical properties by using most of organic acids and inorganic acids commonly used in pharmacy. After continuously experiment, summarizing and improving, the inventors of the present invention have surprisingly found that when using L-proline, in particular to some special preparation conditions, the eutectic crystal of complex of (2S,3R,4R,5S,6R)-2-(4-chloro-3-(4-((3-fluorooxetan-3-yl)methoxy)benzyl)phenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol and bis(L-proline) (hereinafter, referred to as “compound A•L-proline eutectic crystal”) with excellent physical and chemical properties can be obtained. The crystal type of the eutectic crystal can be identified by its characteristic X-ray powder diffraction (XRPD). 
         [0008]    The present invention provides a complex represented by formula I formed by a glucose derivative and L-proline, the complex is composed of compound A and L-proline, a molar ratio of compound A to L-proline is 1:2; and a structure of the complex is as follows: 
         [0000]    
       
                 
         
             
             
         
       
     
         [0009]    wherein, a structure of compound A is as follows: 
         [0000]    
       
                 
         
             
             
         
       
     
         [0010]    The complex refers to an aggregate formed by a series of molecules (e.g. complex organic compounds, inorganic compounds) and simple substance and with a certain (physiological and chemical) functionality or obvious (physical and chemical) characteristics. 
         [0011]    The present invention also provides an eutectic crystal of above complex formed by the glucose derivative and L-proline, 
         [0000]    
       
                 
         
             
             
         
       
     
         [0012]    An X-ray powder diffraction diagram of the eutectic crystal shows characterized peaks at 4.339±0.1, 15.294±0.1, 16.804±0.1, 18.335±0.1, 19.274±0.1, 19.982±0.1, 23.218±0.1 and 24.885±0.1 when diffraction angle is 2θ and under Cu-Kα1 radiation. 
         [0013]    The present invention also provides an eutectic crystal of above complex formed by the glucose derivative and L-proline, 
         [0000]    
       
                 
         
             
             
         
       
     
         [0014]    An X-ray powder diffraction diagram of the eutectic crystal shows characterized peaks at 4.339±0.1, 11.499±0.1, 12.835±0.1, 13.921±0.1, 15.294±0.1, 16.212±0.1, 16.804±0.1, 17.154±0.1, 18.335±0.1, 19.274±0.1, 19.982±0.1, 22.710±0.1, 23.218±0.1, 24.885±0.1, 27.940±0.1, 29.612±0.1 and 30.313±0.1 when diffraction angle is 2θ and under Cu-Kα1 radiation. 
         [0015]    The present invention also provides a preparation method for preparing above eutectic crystal of the complex formed by the glucose derivative and L-proline, which comprises following steps: mixing a solution containing compound A with a solution containing L-proline, cooling and crystallization; 
         [0000]    
       
                 
         
             
             
         
       
     
         [0016]    wherein a solvent of the solution containing L-proline is 95% aqueous ethanol. 
         [0017]    The preparation method preferably further comprises filtering the solution containing compound A with microporous membrane, and then mixing the filtrate with the solution containing L-proline. The microporous membrane is preferably nylon membrane with a pore size of 0.45 μm. 
         [0018]    Usually, the solution containing compound A and the solution containing L-proline is mixed at a temperature of 30° C.-130° C., preferably 50° C.-110° C., more preferably 70° C.-90° C. 
         [0019]    In the preparation method, the solution containing compound A can be prepared by following steps: mixing compound A with a solvent and forming the solution containing compound A. Preferably, compound A and the solvent is mixed under heating. The heating generally refers to make the temperature higher than ambient temperature, which aims to make compound A dissolve in the solvent completely, and generally heating temperature is 30° C.-130° C., preferably 50° C.-110° C., more preferably 70° C.-90° C. 
         [0020]    In the preparation method, the solution containing L-proline can be prepared by following steps: mixing L-proline with 95% aqueous ethanol and forming the solution containing L-proline. Preferably, the amount of the 95% aqueous ethanol is 90 mg/mL-120 mg/mL relative to the mass of L-proline. Wherein, the percentage is a mass percentage. 
         [0021]    A solvent of the solution containing compound A may be with polarity, low toxicity and moderate volatility, which can be easily miscible with 95% aqueous ethanol and is good for dissolving compound A, preferably it is selected from the group consisting of acetone, ethyl acetate, ethanol, water and acetonitrile. In the solution containing compound A, an amount of the solvent which should ensure dissloving compound A is controlled by a concentration of the solution containing compound A generally ranging from 25 mg/mL-400 mg/mL, preferably 50 mg/mL-300 mg/mL. 
         [0022]    In the preparation method, a molar ratio of L-proline to compound A is 1-2. 
         [0023]    The mixing may adopt common methods in the art, such as vortex mixing, magnetic mixing or turbulence mixing etc. The mixing generally lasts 1-30 mins, preferably 1-10 mins. 
         [0024]    The cooling may refer to natural cooling or rapid cooling. The rapid cooling, typically, is to cool the reaction system rapidly by an ice-water bath or an ice-salt bath, etc. 
         [0025]    The cooling which should make the eutectic crystal of the complex formed by compound A and L-proline precipitate is generally cooling to room temperature. In the present invention, the room temperature refers to ambient temperature, which is generally 10° C.-30° C. as defined in pharmacopoeia. 
         [0026]    The rate of the cooling is preferably 1˜20° C./h, more preferably 5˜10° C./h, wherein a high cooling rate accelerates the crystallization process but forms the crystal with small particle size, while a slow cooling rate is conducive to the growth of the crystal and forming a perfect lattice. 
         [0027]    In the preparation method of the present invention, the steps are preferably carried out under stirring. When the cooling and crystallization are carried out under stirring, temperature of the upper, middle and lower portion of the solution can be uniform, which is conducive to the uniformity of the crystal. 
         [0028]    After precipitating, the eutectic crystal of the complex formed by compound A and L-proline can be separated by conventional methods in the art, usually comprising filtering, washing and drying. The filtering can refer to suction filtration. The conditions and procedures of the filtering can refer to conventional filtration operations in the art. The solvent used for the washing is preferably n-heptane. The drying may adopt conventional methods in the art, such as drying at atmospheric pressure or drying under reduced pressure. The temperature of the drying is preferably 40° C. 
         [0029]    The eutectic crystal of the present invention when used as a pharmaceutically active substance is preferably in a substantially pure form, that is substantially free of other forms of crystal fromed by compound A and L-proline. Unless otherwise specified, the present invention also covers a mixture of the eutectic crystal of the complex formed by compound A and L-proline as provided in the present application and one or more other polycrystals of the complex formed by compound A and L-proline. Once the pharmaceutically active substance is a mixed eutectic crystal, more preferably, the mixed eutectic crystal comprises at least 50% of the eutectic crystal of the complex formed by compound A and L-proline provided by the present application, wherein the percentage is a mass fraction. 
         [0030]    The present invention further provides a use of the eutectic crystal of the complex of formula I formed by the glucose derivative and L-proline in preparing a medicament used for treating and/or preventing diseases and/or symptoms induced by the sodium-dependent glucose transporter SGLT (specifically SGLT2). 
         [0031]    In addition, the present invention also provides a use of the eutectic crystal of the complex of formula I formed by the glucose derivative and L-proline in preparing a medicament used for treating and/or preventing metabolic disorders. 
         [0032]    The metabolic disorders generally refer to: type 1 diabetes, type 2 diabetes, diabetic complications, metabolic acidosis or ketoacidosis, reactive hypoglycemia, hyperinsulinemia, glucose metabolic disorders, insulin resistance, metabolic syndrome, dyslipidemia caused by different reasons, atherosclerosis and related diseases, obesity, hypertension, chronic heart failure, edema and hyperuricemia. 
         [0033]    The present invention also provides a use of the eutectic crystal of the complex of formula I formed by the glucose derivative and L-proline in preparing a medicament used for preventing the degeneration of pancreatic β-cells, or a medicament used for improving and/or restoring the functionality of pancreatic β-cells. 
         [0034]    Without departing from the basis of knowledge in the art, each of the above preferred conditions can be any combination to afford the preferred embodiment of the present invention. 
         [0035]    The related materials and reagents in the present invention are commercially available. 
         [0036]    The positive effect of the present invention is that: in the present invention, the complex formed by compound A and L-proline is prepared successfully for the first time. Its eutectic crystal has excellent physical and chemical properties, high water solubility, low hygroscopicity, high stability, which is particularly suitable for preparing various pharmaceutical formulations containing compound A. The preparation method of the present invention is simple and convenient, suitable for industrial production. 
     
    
     
       DESCRIPTION OF FIGURES 
         [0037]      FIG. 1  is the X-ray powder diffraction diagram of the eutectic crystal of the complex formed by compound A and L-proline in effect example 1. 
           [0038]      FIG. 2  is the X-ray powder diffraction diagram of compound A in effect example 1. 
           [0039]      FIG. 3  is the polarizing light microscope photos of the eutectic crystal of the complex formed by compound A and L-proline in effect example 2. 
           [0040]      FIG. 4  is the polarizing light microscope photos of compound A in effect example 2. 
           [0041]      FIG. 5  is the dynamic vapor sorption isotherm of the eutectic crystal of the complex formed by compound A and L-proline in effect example 3. 
           [0042]      FIG. 6  is the dynamic vapor sorption isotherm of compound A in effect example 3. 
       
    
    
     DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT 
       [0043]    The following embodiments further illustrate the present invention, but the present invention is not limited thereto. 
         [0044]    When an experimental condition is not specified in the following examples, conventional methods and conditions can be used, or can be selected from the product manual. 
         [0045]    Compound A in the following example could be prepared with a prior method, such as the method disclosed in WO2012/109996. 
       Example 1 
     Preparation of the Eutectic Crystal of the Complex Formed  by Compound A and L-Proline 
       [0046]    Compound A (25.0 mg) was dissolved in acetone (1 mL) and heated to 30° C. with magnetic stirring until compound A fully dissolved. The solution was filtered with a 0.45 μm microporous membrane (nylon membrane) and the filtrate was placed on a heater at 30° C. An equimolar ratio of 1 mol/L a solution containing L-proline in 95% aqueous ethanol (53.4 μL) was added slowly under magnetic stirring. The solution turned turbid and white solid began to precipitate after 5 minutes magnetic stirring, and then the mixture was cooled down to room temperature at a rate of 10° C./h under stirring, meanwhile a large amount of white precipitate appeared. The precipitated white solid was isolated from the suspension via vacuum filtration, and rinsed with n-heptane once. The product was then placed in a vacuum oven and dried under reduced pressure at 40° C. to obtain the final product. 
       Example 2 
     Preparation of the Eutectic Crystal of the Complex Formed by Compound A and L-Proline 
       [0047]    Compound A (25.0 mg) was dissolved in acetone (1 mL) and heated to 35° C. with magnetic stirring until compound A fully dissolved. The solution was filtered with a 0.45 μm microporous membrane (nylon membrane) and the filtrate was placed on a heater at 35° C. A 2-fold molar ratio of 1 mol/L a solution containing L-proline in 95% aqueous ethanol (106.8 μL) was added slowly under magnetic stirring. The solution turned turbid and white solid began to precipitate after 10 minutes magnetic stirring, and then the mixture was cooled down to room temperature at a rate of 15° C./h under stirring, meanwhile a large amount of white precipitate appeared. The precipitated white solid was isolated from the suspension via vacuum filtration, and rinsed with n-heptane twice. The product was then placed in a vacuum oven and dried under reduced pressure at 40° C. to obtain the final product. 
       Example 3 
     Preparation of the Eutectic Crystal of the Complex Formed by Compound A and L-Proline 
       [0048]    Compound A (25.0 mg) was dissolved in acetonitrile (1 mL) and heated to 30° C. with magnetic stirring until compound A fully dissolved. The solution was filtered with a 0.45 μm microporous membrane (nylon membrane) and the filtrate was placed on a heater at 30° C. An equimolar ratio of 1 mol/L a solution containing L-proline in 95% aqueous ethanol (53.4 μL) was added slowly under magnetic stirring. The solution turned turbid after 10 minutes magnetic stirring, and then the mixture was cooled down to room temperature at a rate of 10° C./h under stirring, meanwhile a large amount of white precipitate appeared. The precipitated white solid was isolated from the suspension via vacuum filtration, and rinsed with n-heptane once. The product was then placed in a vacuum oven and dried under reduced pressure at 40° C. to obtain the final product. 
       Example 4 
     Preparation of the Eutectic Crystal of the Complex Formed by Compound A and L-Proline 
       [0049]    Compound A (25.0 mg) was dissolved in acetonitrile (1 mL) and heated to 45° C. with magnetic stirring until compound A fully dissolved. The solution was filtered with a 0.45 μm microporous membrane (nylon membrane) and the filtrate was placed on a heater at 45° C. A 2-fold molar ratio of 1 mol/L a solution containing L-proline in 95% aqueous ethanol (106.8 μL) was added slowly under magnetic stirring. The solution turned turbid after 15 minutes magnetic stirring, and then the mixture was cooled down to room temperature at a rate of 15° C./h under stirring, meanwhile a large amount of white precipitate appeared. The precipitated white solid was isolated from the suspension via vacuum filtration, and rinsed with n-heptane twice. The product was then placed in a vacuum oven and dried under reduced pressure at 40° C. to obtain the final product. 
       Example 5 
     Preparation of the Eutectic Crystal of the Complex Formed  by Compound A and L-Proline 
       [0050]    Compound A (25.0 mg) was dissolved in ethyl acetate (1 mL) and heated to 40° C. with magnetic stirring until compound A fully dissolved. The solution was filtered with a 0.45 μm microporous membrane (nylon membrane) and the filtrate was placed on a heater at 35° C. An equimolar ratio of 1 mol/L a solution containing L-proline in 95% aqueous ethanol (53.4 μL) was added slowly under magnetic stirring. The solution turned turbid after 10 minutes magnetic stirring, and then the mixture was cooled down to room temperature at a rate of 10° C./h under stirring, meanwhile a large amount of white precipitate appeared. The precipitated white solid was isolated from the suspension via vacuum filtration, and rinsed with n-heptane once. The product was then placed in a vacuum oven and dried under reduced pressure at 40° C. to obtain the final product. 
       Example 6 
     Preparation of the Eutectic Crystal of the Complex Formed by Compound A and L-Proline 
       [0051]    Compound A (25.0 mg) was dissolved in ethyl acetate (1 mL) and heated to 45° C. with magnetic stirring until compound A fully dissolved. The solution was filtered with a 0.45 μm microporous membrane (nylon membrane) and the filtrate was placed on a heater at 45° C. A 2-fold molar ratio of 1 mol/L a solution containing L-proline in 95% aqueous ethanol (106.8 μL) was added slowly under magnetic stirring. The solution turned turbid after 15 minutes magnetic stirring, and then the mixture was cooled down to room temperature at a rate of 15° C./h under stirring, meanwhile a large amount of white precipitate appeared. The precipitated white solid was isolated from the suspension via vacuum filtration, and rinsed with n-heptane twice. The product was then placed in a vacuum oven and dried under reduced pressure at 40° C. to obtain the final product. 
       Example 7 
     Preparation of the Eutectic Crystal of the Complex Formed by Compound A and L-Proline  
       [0052]    Compound A (100.4 mg) was dissolved in acetone (0.5 mL) and heated to 55° C. with magnetic stirring until compound A fully dissolved. The solution was filtered with a 0.45 μm microporous membrane (nylon membrane) and the filtrate was placed on a heater at 55° C. A 2-fold molar ratio of 1 mol/L a solution containing L-proline in 95% aqueous ethanol (429 μL) was added slowly under magnetic stirring. The solution turned turbid and white solid began to precipitate after 20 minutes magnetic stirring, and then the mixture was cooled down to room temperature at a rate of 5° C./h under stirring, meanwhile white precipitate appeared. The precipitated white solid was isolated from the suspension via vacuum filtration, and rinsed with n-heptane three times. The product was then placed in a vacuum oven and dried under reduced pressure at 40° C. to obtain the final product (46.9 mg, yield 31.3%). 
       Example 8 
     Preparation of the Eutectic Crystal of the Complex Formed by Compound A and L-Proline 
       [0053]    Compound A (250.4 mg) was dissolved in acetone (1 mL) and heated to 55° C. with magnetic stirring until compound A fully dissolved. The solution was filtered with a 0.45 μm microporous membrane (nylon membrane) and the filtrate was placed on a heater at 55° C. A 2-fold molar ratio of 1 mol/L a solution containing L-proline in 95% aqueous ethanol (1088 μL) was added slowly under magnetic stirring. The solution turned turbid and white solid began to precipitate after 15 minutes magnetic stirring, and then the mixture was cooled down to room temperature at a rate of 5° C./h under stirring, meanwhile white precipitate appeared. The precipitated white solid was isolated from the suspension via vacuum filtration, and rinsed with n-heptane three times. The product was then placed in a vacuum oven and dried under reduced pressure at 40° C. to obtain the final product (331.8 mg, yield 88.8%). 
       Example 9 
     Preparation of the Eutectic Crystal of the Complex Formed  by Compound A and L-Proline 
       [0054]    Compound A (250.9 mg) was dissolved in acetone (1 mL) and heated to 55° C. with magnetic stirring until compound A fully dissolved. The solution was filtered with a 0.45 μm microporous membrane (nylon membrane) and the filtrate was placed on a heater at 55° C. A 2-fold molar ratio of 1 mol/L a solution containing L-proline in 95% aqueous ethanol (1090 μL) was added slowly under magnetic stirring. The solution turned turbid and white solid began to precipitate after 25 minutes magnetic stirring, and then the mixture was cooled down to room temperature at a rate of 5° C./h under stirring, meanwhile white precipitate appeared. The precipitated white solid was isolated from the suspension via vacuum filtration, and rinsed with n-heptane three times. The product was then placed in a vacuum oven and dried under reduced pressure at 40° C. to obtain the final product (271.0 mg, yield 72.5%). 
       Example 10 
     Preparation of the Eutectic Crystal of the Complex Formed by Compound A and L-Proline 
       [0055]    Compound A (250.9 mg) was dissolved in acetone (1 mL) and heated to 55° C. with magnetic stirring until compound A fully dissolved. The solution was filtered with a 0.45 μm microporous membrane (nylon membrane) and the filtrate was placed on a heater at 55° C. A 2-fold molar ratio of 1 mol/L a solution containing L-proline in 95% aqueous ethanol (1090 μL) was added slowly under magnetic stirring. The solution turned turbid and white solid began to precipitate after 25 minutes magnetic stirring, and then the mixture was cooled down to room temperature at a rate of 5° C./h under stirring, meanwhile white precipitate appeared. The precipitated white solid was isolated from the suspension via vacuum filtration, and rinsed with n-heptane three times. The product was then placed in a vacuum oven and dried under reduced pressure at 40° C. to obtain the final product (271.0 mg, yield 72.5%). 
       Example 11 
     Preparation of the Eutectic Crystal of the Complex Formed by Compound A and L-Proline 
       [0056]    Compound A (100.7 mg) was dissolved in acetone (0.5 mL) and heated to 55° C. with magnetic stirring until compound A fully dissolved. The solution was filtered with a 0.45 μm microporous membrane (nylon membrane) and the filtrate was placed on a heater at 55° C. A 2-fold molar ratio of 1 mol/L a solution containing L-proline in 95% aqueous ethanol (433 μL) was added slowly under magnetic stirring. The solution turned turbid and white solid began to precipitate after 25 minutes magnetic stirring, and then the mixture was cooled down to room temperature at a rate of 5° C./h undre stirring, meanwhile white precipitate appeared. The precipitated white solid was isolated from the suspension via vacuum filtration, and rinsed with n-heptane three times. The product was then placed in a vacuum oven and dried under reduced pressure at 40° C. to obtain the final product (93.6 mg, yield 64.4%). 
       Example 12 
     Preparation of the Eutectic Crystal of the Complex Formed by Compound A and L-Proline 
       [0057]    Compound A (100.8 mg) was dissolved in acetone (0.5 mL) and heated to 55° C. with magnetic stirring until compound A fully dissolved. The solution was filtered with a 0.45 μm microporous membrane (nylon membrane) and the filtrate was placed on a heater at 55° C. A 2-fold molar ratio of 1 mol/L a solution containing L-proline in 95% aqueous ethanol (433 μL) was added slowly under magnetic stirring. The solution turned turbid and white solid began to precipitate after 25 minutes magnetic stirring, and then the mixture was cooled down to room temperature at a rate of 5° C./h under stirring, meanwhile white precipitate appeared. The precipitated white solid was isolated from the suspension via vacuum filtration, and rinsed with n-heptane three times. The product was then placed in a vacuum oven and dried under reduced pressure at 40° C. to obtain the final product (107.8 mg, yield 71.4%). 
       Example 13 
     Preparation of the Eutectic Crystal of the Complex Formed by Compound A and L-Proline 
       [0058]    Compound A (200.4 mg) was dissolved in acetone (1.2 mL) and heated to 55° C. with magnetic stirring until compound A fully dissolved. The solution was filtered with a 0.45 μm microporous membrane (nylon membrane) and the filtrate was placed on a heater at 55° C. A 2-fold molar ratio of 1 mol/L a solution containing L-proline in 95% aqueous ethanol (856 μL) was added slowly under magnetic stirring. The solution turned turbid and white solid began to precipitate after 25 minutes magnetic stirring, and then the mixture was cooled quickly with an ice-bath for 10 mins and white precipitate appeared. The precipitated white solid was isolated from the suspension via vacuum filtration, and rinsed with n-heptane three times. The product was then placed in a vacuum oven and dried under reduced pressure at 40° C. to obtain the final product. 
       Example 14 
     Preparation of the Eutectic Crystal of the Complex Formed by Compound A and L-Proline 
       [0059]    Compound A (272.3 mg) was dissolved in acetone (3.6 mL) and heated to 55° C. with magnetic stirring until compound A fully dissolved. The solution was filtered with a 0.45 μm microporous membrane (nylon membrane) and the filtrate was placed on a heater at 55° C. A 2-fold molar ratio of 1 mol/L a solution containing L-proline in 95% aqueous ethanol (1164 μL) was added slowly under magnetic stirring. The solution turned turbid and white solid began to precipitate after 10 minutes magnetic stirring, and then the mixture was cooled down to room temperature at a rate of 5° C./h under stirring, meanwhile white precipitate appeared. The precipitated white solid was isolated from the suspension via vacuum filtration, and rinsed with n-heptane three times. The product was then placed in a vacuum oven and dried under reduced pressure at 40° C. to obtain the final product (327.8 mg, yield 80.6%). 
       Example 15 
     Preparation of the Eutectic Crystal of the Complex Formed by Compound A and L-Proline 
       [0060]    Compound A (400.6 mg) was dissolved in acetone (4.8 mL) and heated to 55° C. with magnetic stirring until compound A fully dissolved. The solution was filtered with a 0.45 μm microporous membrane (nylon membrane) and the filtrate was placed on a heater at 55° C. A 2-fold molar ratio of 1 mol/L a solution containing L-proline in 95% aqueous ethanol (1712 μL) was added slowly under magnetic stirring. The solution turned turbid after 10 minutes magnetic stirring and a lot of white solid precipitated after another 0.5 min, and then the mixture was cooled down to room temperature at a rate of 20° C./h under magnetic stirring. The precipitated white solid was isolated from the suspension via vacuum filtration, and rinsed with n-heptane three times. The product was then placed in a vacuum oven and dried under reduced pressure at 40° C. to obtain the final product (461 mg, yield 77.1%). 
       Example 16 
     Preparation of the Eutectic Crystal of the Complex Formed by Compound A and L-Proline 
       [0061]    Compound A (589 mg) was dissolved in acetone (7.068 mL) and heated to 55° C. with magnetic stirring until compound A fully dissolved. The solution was filtered with a 0.45 μm microporous membrane (nylon membrane) and the filtrate was placed on a heater at 55° C. A 2-fold molar ratio of 1 mol/L a solution containing L-proline in 95% aqueous ethanol (2977 μL) was added slowly under magnetic stirring. The solution turned turbid after 1 minute magnetic stirring and a lot of white solid began to precipitate, and then the mixture was cooled down to room temperature at a rate of 20° C./h under magnetic stirring. The precipitated white solid was isolated from the suspension via vacuum filtration, and rinsed with n-heptane three times. The product was then placed in a vacuum oven and dried under reduced pressure at 40° C. to obtain the final product. 
       Example 17 
     Preparation of the Eutectic Crystal of the Complex Formed by Compound A and L-Proline 
       [0062]    Compound A (407.52 mg) was dissolved in acetone (4.8 mL) and heated to 55° C. with magnetic stirring until compound A fully dissolved. The solution was filtered with a 0.45 μm microporous membrane (nylon membrane) and the filtrate was placed on a heater at 55° C. A 2-fold molar ratio of 1 mol/L a solution containing L-proline in 95% aqueous ethanol (1742 μL) was added slowly under magnetic stirring. The solution turned turbid after 1 minute magnetic stirring and a lot of white solid began to precipitate after another 1 minute, and then the mixture was cooled down to room temperature at a rate of 10° C./h under magnetic stirring. The precipitated white solid was isolated from the suspension via vacuum filtration, and rinsed with n-heptane three times. The product was then placed in a vacuum oven and dried under reduced pressure at 40° C. to obtain the final product (476.75 mg, yield 78.4%). 
       Example 18 
     Preparation of the Eutectic Crystal of the Complex Formed by Compound A and L-Proline 
       [0063]    Compound A (410.5 mg) was dissolved in acetone (4.8 mL) and heated to 55° C. with magnetic stirring until compound A fully dissolved. The solution was filtered with a 0.45 μm microporous membrane (nylon membrane) and the filtrate was placed on a heater at 55° C. A 2-fold molar ratio of 1 mol/L a solution containing L-proline in 95% aqueous ethanol (1754 μL) was added slowly under magnetic stirring. The solution turned turbid after 1 minute magnetic stirring and a lot of white solid began to precipitate after another 1 min, and then the mixture was cooled down to room temperature at a rate of 10° C./h under magnetic stirring. The precipitated white solid was isolated from the suspension via vacuum filtration, and rinsed with n-heptane three times. The product was then placed in a vacuum oven and dried under reduced pressure at 40° C. to obtain the final product (445.98 mg, yield 72.8%). 
       Example 19 
     Preparation of the Eutectic Crystal of the Complex Formed by Compound A and L-Proline 
       [0064]    Compound A (404.52 mg) was dissolved in acetone (4.8 mL) and heated to 55° C. with magnetic stirring until compound A fully dissolved. The solution was filtered with a 0.45 μm microporous membrane (nylon membrane) and the filtrate was placed on a heater at 55° C. A 2-fold molar ratio of 1 mol/L a solution containing L-proline in 95% aqueous ethanol (1729 μL) was added slowly under magnetic stirring. The solution turned turbid after 1 minute magnetic stirring and a lot of white solid began to precipitate, and then the mixture was cooled down to room temperature at a rate of 20° C./h under magnetic stirring. The precipitated white solid was isolated from the suspension via vacuum filtration, and rinsed with n-heptane three times. The product was then placed in a vacuum oven and dried under reduced pressure at 40° C. to obtain the final product (437.95 mg, yield 72.6%). 
       Comparative Example 1 
     Preparation of the Eutectic Crystal of the Complex Formed by Compound A and L-Proline 
       [0065]    Compound A (102.0 mg) was dissolved in isopropyl alcohol (0.5 mL) and heated to 80° C. with magnetic stirring until compound A fully dissolved. The solution was filtered with a 0.45 μm microporous membrane (nylon membrane) and the filtrate was placed on a heater at 80° C. A 2-fold molar ratio of 1 mol/L a solution containing L-proline in 95% aqueous ethanol (436 μL) was added slowly under magnetic stirring. The solution was clear after 1 h magnetic stirring, and then the mixture was cooled down to room temperature at a rate of 10° C./h under magnetic stirring. The solution turned turbid and little amount of white solid began to precipitate after stirring for 12 hours at room temperature. However, the mixture became clear when stirring stopped and small amount of oil was observed floating on the surface of the solution. There was no solid observed after the mixture was stored at 4° C. for 12 hours. 
         [0066]    From the above comparative example 1, the eutectic crystal of the complex formed by compound A and L-proline could not be obtained if isopropyl alcohol was used as a solvent for dissolving compound A, even other conditions strictly followed the previously described preparation method in the present invention. It proved that the solvent for dissolving compound A was a key factor for the preparation method. The solvent should be not only a good solvent for dissolving compound A, but also a solvent miscible with 95% aqueous ethanol. 
       Effect Example 1 
     Identification of X-Ray Powder Diffraction 
       [0067]    1. Samples: Eutectic crystal of the complex formed by compound A and L-proline prepared in example 1-19 and compound A. 
         [0068]    2. Parameters of X-ray powder diffraction: A Cu-Kα 1  source (=1.54056 angstrom); Operating Voltage: 40 kV; Operating Current: 40 mA; Detector: PSD Detector; Scans Angle: 4-40 degrees (2-theta); Step value: 0.05°; Scanning Speed: 0.5 sec/step. 
         [0069]    3. Experimental Result 
         [0070]    The X-ray powder diffraction diagram of the eutectic crystal of the complex formed by compound A and L-proline prepared in example 1-19 is shown in  FIG. 1 . It can be seen from  FIG. 1  that the diffraction peaks of the eutectic crystal of the complex formed by compound A and L-proline were selected from the following peaks: diffraction angle 2θ=4.339, 11.499, 12.835, 13.921, 15.294, 16.212, 16.804, 17.154, 18.335, 19.274, 19.982, 22.710, 23.218, 24.885, 27.940, 29.612 and 30.313 degrees. Peaks corresponding to marked numerals in  FIG. 1  are shown in Table 1: 
         [0000]    
       
         
               
               
               
             
               
               
               
             
           
               
                   
                 TABLE 1 
               
               
                   
                   
               
               
                   
                 Peak number 
                 2θ value 
               
               
                   
                   
               
             
             
               
                   
               
             
          
           
               
                   
                 1 
                 4.339 
               
               
                   
                 2 
                 11.499 
               
               
                   
                 3 
                 12.835 
               
               
                   
                 4 
                 13.921 
               
               
                   
                 5 
                 15.294 
               
               
                   
                 6 
                 16.212 
               
               
                   
                 7 
                 16.804 
               
               
                   
                 8 
                 17.154 
               
               
                   
                 9 
                 18.335 
               
               
                   
                 10 
                 19.274 
               
               
                   
                 11 
                 19.982 
               
               
                   
                 12 
                 22.710 
               
               
                   
                 13 
                 23.218 
               
               
                   
                 14 
                 24.885 
               
               
                   
                 15 
                 27.940 
               
               
                   
                 16 
                 29.612 
               
               
                   
                 17 
                 30.313 
               
               
                   
                   
               
             
          
         
       
     
         [0071]    The X-ray powder diffraction diagram of starting material compound A is shown in  FIG. 2 . It can be seen from  FIG. 2  that there is no obvious diffraction peaks in the X-ray powder diffraction diagram of starting material compound A, which means that starting material compound A is amorphous. 
       Effect Example 2 
     Polarized Light Microscopy 
       [0072]    1. Samples: Eutectic crystal of the complex formed by compound A and L-proline prepared in example 1-19 and compound A. 
         [0073]    2. Parameters of Polarized light microscopy: Eyepiece 10×, Objective 10×. 
         [0074]    3. Experimental Result 
         [0075]    The polarizing light microscope photo of the eutectic crystal of the complex formed by compound A and L-proline prepared in example 1-19 is shown in  FIG. 3 . It can be seen from  FIG. 3  that the eutectic crystal has obvious birefringence and shows a crystal habit of irregular particle. The particle size distributes in the range from 10 μm to 50 μm. 
         [0076]    The polarizing light microscope photo of starting material compound A is shown in  FIG. 4 . Compound A has no birefringence phenomenon and is in irregular glass state. 
         [0077]    It can be seen from above comparison that comparing with the amorphous compound A, the eutectic crystal of the complex formed by compound A and L-proline of the present invention with a particle form has better stability and is conducive to the process for pharmaceutical preparation. 
       Effect Example 3 
     Dynamic Vapor Sorption Test 
       [0078]    1. Samples: Eutectic crystal of the complex formed by compound A and L-proline prepared in example 1-19 and compound A. 
         [0079]    2. Parameters of dynamic vapor sorption test: The instrument: a dynamic moisture absorption analyzer (DVS Advantage, Surface Measurement System Ltd); Experimental temperature: 25° C.; Adsorption range: 0-95% relative humidity; Step interval: 5% relative humidity; Balance standard of weight gain: less than 0.01% weight change in 5 minutes; Longest time of balance: 120 minutes. 
         [0080]    3. Experimental Result 
         [0081]    The dynamic vapor sorption isotherm of the eutectic crystal of the complex formed by compound A and L-proline prepared in example 1-19 is shown in  FIG. 5 . It can be seen in  FIG. 5  that when an initial mass is 10.2006 mg and relative humidity is increased from 0% to 85% RH, L-proline eutectic crystal absorbs moisture slowly and the moisture absorbed is only 2.174%, while RH is 95%, the weight increased finally is about 9.967%, which indicates that the eutectic crystal of the complex formed by compound A and L-proline is not very sensitive to moisture under normal conditions. In  FIG. 5 , Y axis is the weight gain percentage of the sample against its dry weight. In  FIG. 5 , relative humidity is defined as the ratio of the vapor pressure of water in air to the saturated vapor pressure of water. 
         [0082]    The dynamic vapor sorption isotherm of compound A is shown in  FIG. 6 . It can be seen in  FIG. 6  that when an initial mass is 10.3109 mg and relative humidity is 50%, the moisture absorption rate of compound A significantly accelerates, the moisture absorbed is about 4.8%, and it keeps absorbing moisture rapidly, the weight increased finally is 14.72% when the relative humidity reaches 95% RH. Thus, compound A is sensitive to moisture. In  FIG. 6 , Y axis is the weight gain percentage of the sample against its dry weight. In  FIG. 6 , relative humidity is defined as the ratio of the vapor pressure of water in air to the saturated vapor pressure of water. 
         [0083]    It can be seen from above comparison that the eutectic crystal of the complex formed by compound A and L-proline of the present invention has low hygroscopicity, which shows obvious advantages. 
       Effect Example 4 
     Solubility Test 
       [0084]    1. Samples: Eutectic crystal of the complex formed by compound A and L-proline prepared in example 1-19 and compound A. 
         [0085]    2. Solubility test method: 2-3 mg sample was accurately weighed and put into a small vial, the right amount of ultra-pure water was added to make a target concentration of 2.0 mg/mL, the vial was rotated for 18 hrs at 25° C. till no changes of solubility, HPLC was used to determine the drug concentration, the solubility of the drug was calculated by preparing standard curve. 
         [0086]    HPLC measurement condition: Instrument: Agilent 1200 HPLC; Chromatographic column: Zorbax SB-C8 (3.5 μm, 4.6×75 mm), SN: USEB009791; Mobile phase A: 10 mmol/L aqueous ammonium acetate solution (0.77 g of ammonium acetate was mixed evenly with 1 L of Milli-Q water), Mobile phase B: Acetonitrile, Mobile phase A: Mobile phase B=65:35 (v:v); Column temperature: 25° C.; Wave length: 220 nm; Sampling volume: 10 μL; Flow velocity: 1 mL/min; Detection time: 5 min, t 0 =0.65 min, t R =2.7 min, K′=3.15 (capacity factor, this value should be greater than 2), tailing factor=1.1. 
         [0087]    3. Experimental Result 
         [0000]    
       
         
               
             
               
               
               
               
               
               
               
             
           
               
                 TABLE 2 
               
             
             
               
                   
               
               
                 solubility of Eutectic crystal of the compound A and L-proline in 
               
               
                 different aqueous medium 
               
             
          
           
               
                   
                   
                   
                 Target 
                   
                   
                 Solubility 
               
               
                   
                   
                   
                 concen- 
                   
                 pH 
                 measured 
               
               
                   
                 Mass 
                 Volume 
                 tration 
                 Visual 
                 (after 
                 by HPLC 
               
               
                 Medium 
                 (mg) 
                 (mL) 
                 (mg/mL) 
                 solubility 
                 filtration) 
                 (mg/mL) 
               
               
                   
               
               
                 Water 
                 2.113 
                 1.056 
                 2.000 
                 clear 
                 7.941 
                 1.94 
               
               
                 0.1N 
                 1.882 
                 0.941 
                 2.000 
                 clear 
                 0.975 
                 1.91 
               
               
                 HCl 
               
               
                 pH 2 
                 1.724 
                 0.862 
                 2.000 
                 clear 
                 1.963 
                 1.93 
               
               
                 pH 4 
                 2.162 
                 1.080 
                 2.000 
                 clear 
                 4.610 
                 1.93 
               
               
                 pH 6 
                 1.977 
                 0.988 
                 2.000 
                 clear 
                 6.481 
                 1.93 
               
               
                 pH 8 
                 1.714 
                 0.857 
                 2.000 
                 clear 
                 8.060 
                 1.93 
               
               
                 SGF 
                 1.638 
                 0.819 
                 2.000 
                 clear 
                 2.871 
                 1.85 
               
               
                 SIF- 
                 1.758 
                 0.879 
                 2.000 
                 clear 
                 6.533 
                 1.98 
               
               
                 Fasted 
               
               
                 SIF-Fed 
                 2.057 
                 1.028 
                 2.000 
                 clear 
                 4.956 
                 2.08 
               
               
                   
               
             
          
         
       
     
         [0000]                                                  TABLE 3                   solubility of compound A in different aqueous medium                        Target           Solubility                   concen-       pH   measured           Mass   Volume   tration   Visual   (after   by HPLC       Medium   (mg)   (mL)   (mg/mL)   solubility   filtration)   (mg/mL)               Water   3.102   1.552   2.000   clear   8.540   1.68       0.1N   3.190   1.596   2.000   clear   2.182   1.56       HCl       pH 2   2.952   1.476   2.000   clear   2.151   1.61       pH 4   3.298   1.648   2.000   clear   5.094   1.64       pH 6   2.802   1.400   2.000   clear   6.180   1.64       pH 8   3.194   1.596   2.000   clear   8.020   1.57       SGF   2.634   1.316   2.000   clear   2.178   1.91       SIF-   2.776   1.388   2.000   clear   6.146   2.04       Fasted       SIF-Fed   3.433   1.716   2.000   clear   5.099   1.92                    
Wherein, SGF refers to manual mode gastric juice, SIF-Fasted refers to simulation intestinal juice (before meal), SIF-Fed refers to simulation intestinal juice (after meal).