CN114644642B - Crystal form A of thienopyridine compound, preparation method and pharmaceutical composition thereof - Google Patents

Abstract

The present invention discloses a compound 2- (4- (7- (2-fluoro-4- (1- ((4-fluorophenyl) carbamoyl) cyclopropane-1-carboxamide) phenoxy) thieno [3, 2-b)]Crystalline form a of pyridin-2-yl) -1H-pyrazol-1-yl) ethyl valine ester hydrochloride, methods of preparation, and pharmaceutical compositions thereof. The crystal form has high chemical stability, high temperature resistance, high humidity resistance, good solubility and high bioavailability, and the pharmaceutical composition has good dissolution rate, thus being applicable to the development of preparations. The structural formula of the compound is shown as a formula (I):

Description

Crystal form A of thienopyridine compound, preparation method and pharmaceutical composition thereof

Technical Field

The invention belongs to the technical field of medicines, and particularly relates to a novel crystal form of a thienopyridine compound, a preparation method thereof and a pharmaceutical composition containing the crystal form of the thienopyridine compound.

Background

The same substance has two or more spatial arrangements and unit cell parameters, and a phenomenon in which a plurality of crystal forms are formed is called polymorphism (polymorphism). Many crystalline drugs have polymorphism, and different crystal forms of the same drug may be significantly different in appearance, solubility, melting point, dissolution rate, bioavailability and the like, thereby affecting the stability, bioavailability and curative effect of the drug. The polymorphism of drugs is one of the important factors affecting the quality and clinical efficacy of drugs, so that the analysis of the crystal forms of drugs with polymorphism is of particular interest when developing drugs with polymorphism.

Chinese patent application CN113336768A discloses a series of potential thienopyridine-based small molecule anticancer compounds including the compound 2- (4- (7- (2-fluoro-4- (1- ((4-fluorophenyl) carbamoyl) cyclopropane-1-carboxamide) phenoxy) thieno [3,2-b ] pyridin-2-yl) -1H-pyrazolyl-1-yl) ethyl valine ester hydrochloride. The structural formula of the compound is shown as formula (I).

Although this patent application discloses a method for preparing the compound, no disclosure shows that any compound can be stably crystallized or purified. On the basis, the invention further researches the new crystal forms of the thienopyridine micromolecule anticancer compound and the influence of the new crystal forms on the stability, the solubility, the bioavailability, the dissolution rate and the like of the medicine.

Disclosure of Invention

An object of the present invention is to provide a novel crystalline form of a compound, namely, crystalline form a of 2- (4- (7- (2-fluoro-4- (1- ((4-fluorophenyl) carbamoyl) cyclopropane-1-carboxamide) phenoxy) thieno [3,2-b ] pyridin-2-yl) -1H-pyrazolyl-1-yl) ethyl valine ester hydrochloride, the structural formula of which is shown in formula (I).

The novel crystal form has high chemical stability, high temperature resistance, high humidity resistance, good solubility and high bioavailability, and the pharmaceutical composition has good dissolution rate, thus being applicable to the development of preparations.

The compound A crystal form is characterized by X-ray powder diffraction (XRPD), differential Scanning Calorimetry (DSC), thermogravimetric analysis (TG) and infrared spectrum (IR), and has the properties required for preparing a pharmaceutical preparation.

It is another object of the present invention to provide a process for the preparation of the novel crystalline form of the compound.

It is a further object of the present invention to provide pharmaceutical compositions containing the crystalline forms of the compounds.

According to one aspect of the invention, a crude product of the compound (shown as a formula (I)) is prepared, and then the crude product of the compound is crystallized and transcrystalline by a recrystallization and transcrystalline method to obtain a compound crystal form.

The obtained crystals were confirmed to be a novel crystal called form a by detection and analysis of the substance by X-ray powder diffraction (XRPD), differential Scanning Calorimetry (DSC), thermogravimetric analysis (TG), infrared spectroscopy (IR), and the like.

Specifically, when X-ray powder diffraction is performed with Cu-K alpha, the X-ray powder diffraction pattern of the compound A crystal form has characteristic diffraction peaks at the following 2 theta DEG positions: the relative intensities of these peaks (I/I) were 8.01±0.20°, 10.38±0.20°, 10.86±0.20°, 12.99±0.20°, 13.44±0.20°, 14.94±0.20°, 16.02±0.20°, 17.76±0.20°, 18.83±0.20°, 20.11±0.20 °, 20.36±0.20±22.20±0.20°, 23.13±0.20°, 24.70±0.20 °, 24.92±0.20±25.40±0.20°, 26.72 ±0.20 °, 27.89 ±0.20°, 28.94±0.20° ₀ ) All greater than or equal to 30%.

The X-ray powder diffraction pattern of the compound A crystal form also has characteristic diffraction peaks at the following 2 theta DEG positions: the relative intensities (I/I0) of the peaks are greater than or equal to 15% at 6.29+ -0.20 °, 9.55+ -0.20 °, 12.55+ -0.20 °, 14.10+ -0.20 °, 14.30+ -0.20 °, 17.19+ -0.20 °, 19.16+ -0.20 °, 22.43+ -0.20 °, 26.11+ -0.20 °, 29.51+ -0.20 °.

The X-ray powder diffraction pattern of the compound A crystal form also has characteristic diffraction peaks at the following 2 theta DEG positions: 15.42.+ -. 0.20 °, 23.79.+ -. 0.20 °, 24.15.+ -. 0.20 °, 30.65.+ -. 0.20 °, 31.09.+ -. 0.20 °, 32.56.+ -. 0.20 °, 36.34.+ -. 0.20 °, 38.68.+ -. 0.20 °, relative intensities of these peaks (I/I ₀ ) Are all greater than or equal to 5% (see figure 1).

The compound A crystal form is characterized by an X-ray powder diffraction pattern, and the relative intensity of characteristic diffraction peaks is close to the following values.

TABLE 1X-ray powder diffraction pattern data for Compound A crystalline form

The term "proximity" herein refers to the uncertainty of the relative intensity measurements. Those skilled in the art understand that the uncertainty in the relative intensities is very dependent on the measurement conditions. The relative intensity values may vary, for example, within a range of 25% or preferably within a range of 10%.

The compound A crystal form of the invention has an X-ray powder diffraction pattern shown in figure 1.

The invention adopts differential scanning calorimetric analysis (DSC) technology to characterize the compound A crystal form (see figure 2), and the differential scanning calorimetric diagram shows that two endothermic peaks exist at 111.1 ℃ and 197.4 ℃ of a sample.

The invention adopts the thermogravimetric analysis (TG) technology to characterize the crystal form A of the compound (see figure 3), and the thermogravimetric analysis map shows that the weight loss of the test sample is 2.55% in the range of room temperature to 136.0 ℃; a loss of weight in the range of 136.0 ℃ to 204.0 ℃ of 1.32%; a loss of weight in the range 204.0 ℃ to 481.0 ℃ of 53.58%; the loss weight is 42.31% in the range of 481.0 ℃ to 700.0 ℃. Indicating that the compound degraded with increasing temperature.

The invention adopts infrared spectrum (IR) technology to characterize the A crystal form (see figure 4), and the infrared spectrum shows that the sample is 3356cm ^-1 、3070cm ^-1 、3027cm ^-1 、2963cm ^-1 、2879cm ^-1 、2785cm ^-1 、2683cm ^-1 、2618cm ^-1 、1752cm ^-1 、1693cm ^-1 、1642cm ^-1 、1602cm ^-1 、1556cm ^-1 、1507cm ^-1 、1495cm ^-1 、1469cm ^-1 、1407cm ^-1 、1327cm ^-1 、1288cm ^-1 、1251cm ^-1 、1200cm ^-1 、868cm ^-1 、829cm ^-1 There is a strong absorption peak.

According to another aspect of the invention, the process for preparing form a comprises the steps of: adding crude 2- (4- (7- (2-fluoro-4- (1- ((4-fluorophenyl) carbamoyl) cyclopropane-1-carboxamide) phenoxy) thieno [3,2-b ] pyridin-2-yl) -1H-pyrazolyl-1-yl) ethyl valine ester hydrochloride into tetrahydrofuran, ethyl acetate, C1-C4 alkyl alcohol, C3-C4 alkyl ketone or a mixed solvent thereof and water, heating and refluxing until the crude product is dissolved; and (3) cooling the solution after clarification until solid is precipitated, filtering and collecting the solid, drying the solid, transferring the obtained solid into methanol for crystallization, and finally drying by blowing to obtain the crystal form A. The solvent is preferably tetrahydrofuran; the alcohol is selected from methanol, ethanol, propanol and isopropanol, preferably methanol; the ketone is selected from acetone, methyl ethyl ketone, etc., preferably acetone. Tetrahydrofuran, ethyl acetate, C1-C4 alkyl alcohol, C3-C4 alkyl ketone to water in a volume ratio (V/V) of 3:1 to 15:1, preferably 9:1; the ratio of the crude product to the solvent is calculated according to g/mL, and the weight volume ratio is 1: 4-20, tetrahydrofuran is preferably 1:10, and methanol is preferably 1:8. The solution is preferably heated to 50-90 ℃, more preferably tetrahydrofuran to 80 ℃, and methanol to 70 ℃; according to this embodiment, the solid is allowed to stand for 2 to 16 hours, more preferably 12 hours. The precipitation temperature is 0 to 30 ℃, preferably 15 to 25 ℃. Filtering after solid separation is completed, adding methanol for pulping, and performing crystal transformation operation; the weight-volume ratio of the solid obtained by recrystallization to the methanol is 1:1 to 10, preferably 1:2; the pulping temperature is 10-50 ℃, preferably 20-30 ℃; the beating time is 1 to 10 hours, more preferably 4 hours. After the crystal transformation is finished, the obtained suspension is filtered to obtain white powder, and the white powder is dried, wherein the drying temperature is 30-70 ℃, preferably 50 ℃.

According to a further aspect of the present invention there is provided a pharmaceutical composition comprising the crystalline form a above and optionally a pharmaceutically acceptable pharmaceutical excipient.

The pharmaceutical compositions may further be formulated in a pharmaceutically acceptable form, including orally or parenterally, according to conventional formulation methods. In a pharmaceutically acceptable form, a therapeutically effective amount of form a should be included. By "therapeutically effective amount" is meant that at such doses, the compounds of the invention are capable of ameliorating or alleviating symptoms of the disease, or of inhibiting and blocking the progression of the disease.

The term "pharmaceutical excipients" refers to the various carriers and/or excipients used in the manufacture and formulation of pharmaceutical products, and is all matter contained in the pharmaceutical formulation except for the active ingredient.

According to a further aspect of the invention, the crystalline forms of the invention may be used alone in the preparation of a medicament for the treatment of a cell proliferative disorder, or may be prepared in combination with other therapeutic agents to act synergistically.

Crystalline form a of 2- (4- (7- (2-fluoro-4- (1- ((4-fluorophenyl) carbamoyl) cyclopropane-1-carboxamide) phenoxy) thieno [3,2-B ] pyridin-2-yl) -1H-pyrazolyl-1-yl) ethyl valine ester hydrochloride of the present invention for use in the treatment of cell proliferative disorders, primarily cancer, including but not limited to non-small cell lung cancer, progressive large cell lymphoma, liver cancer, stomach cancer, colorectal cancer, breast cancer, pancreatic cancer, ovarian cancer, diffuse large B-cell lymphoma, glioma, esophageal cancer, brain cancer or neck cancer.

The crystal form A of 2- (4- (7- (2-fluoro-4- (1- ((4-fluorophenyl) carbamoyl) cyclopropane-1-carboxamide) phenoxy) thieno [3,2-b ] pyridine-2-yl) -1H-pyrazolyl-1-yl) ethyl valine ester hydrochloride prepared by the invention has the characteristics of high chemical stability, high temperature resistance and high humidity resistance, has the properties required by preparation, is simple and convenient to produce, convenient to store and easier to control the quality, and has the dissolution rate of more than 80% in 15 minutes as proved by experiments.

Drawings

FIG. 1 is an X-ray powder diffraction pattern of novel form A obtained in example 1 of the present invention;

FIG. 2 is a Differential Scanning Calorimetric (DSC) profile of the novel form A obtained in example 1 of the present invention;

FIG. 3 is a thermogravimetric analysis (TG) plot of the novel form A obtained in example 1 of the present invention;

FIG. 4 is an Infrared (IR) spectrum of novel form A obtained in example 1 of the present invention;

FIG. 5 is an HPLC chart of novel form A obtained in example 1 of the present invention.

FIG. 6 is a graph of blood concentration versus time (dose 200 mg/kg) of td32-4 obtained by in vivo conversion of the novel form A obtained in example 1 with crude product of the present invention.

Detailed Description

The raw materials and the reagents used in the invention are all commercial products.

Preparing a crude product:

preparation of crude 2- (4- (7- (2-fluoro-4- (1- ((4-fluorophenyl) carbamoyl) cyclopropane-1-carboxamide) phenoxy) thieno [3,2-b ] pyridin-2-yl) -1H-pyrazolyl-1-yl) ethyl valine ester hydrochloride starting from td32-4 (N-3-fluoro 4- ((2- (1- (2-hydroxymethyl) -1H-pyrazol-4-yl) thieno [3,2-b ] pyridin-7-yl) oxy) phenyl) -N- (4-fluorophenyl) cyclopropane-1, 1-dicarboxamide by the method of the patent application (patent application number CN 202010099747.2).

Example 1

100g of crude 2- (4- (7- (2-fluoro-4- (1- ((4-fluorophenyl) carbamoyl) cyclopropane-1-carboxamide) phenoxy) thieno [3,2-b ] pyridin-2-yl) -1H-pyrazolyl-1-yl) ethyl valine ester hydrochloride are weighed, added to a reaction flask, 1000mL of a mixed solution of tetrahydrofuran and water (Vtetrahydrofuran: vwater=9:1) are added, and the mixture is heated to reflux to 80℃with stirring. Stirring for 30min after dissolution, cooling to 15-25 ℃, stirring for crystallization for 12h, filtering, and drying a filter cake to obtain 80g of solid; adding the obtained solid into a reaction bottle, adding 160ml of methanol, pulping for 4 hours at 20-25 ℃, and carrying out suction filtration; the filter cake obtained was air-dried at 50℃to give 76g of white powder, 76% of the total yield in the two steps. The obtained compound is 2- (4- (7- (2-fluoro-4- (1- ((4-fluorophenyl) carbamoyl) cyclopropane-1-carboxamide) phenoxy) thieno [3,2-b ] pyridin-2-yl) -1H-pyrazolyl-1-yl) ethyl valine ester hydrochloride in the A crystal form, and the property identification is shown in figures 1-5.

Examples sample test conditions:

x-ray powder diffraction (XRPD):

detection instrument: sharp image (Empyrean) X-ray diffractometer

Detection conditions: cu target K alpha rays, voltage 40kV, current 40mA, emission slit 1/8 degrees, anti-scattering slit 1/4 degrees, anti-scattering slit 7.5mm,2 theta degree range: 3-60 deg., step size of 0.02 deg., dwell time of 40s per step.

The detection basis is as follows: detection results of four 0451X-ray diffraction methods of the pharmacopoeia of the people's republic of China (2020 edition): as in fig. 1.

(II) Differential Scanning Calorimeter (DSC):

detection instrument: DSC 214 differential scanning calorimeter of NETZSCH company of Germany

Detection conditions:

atmosphere: n2, 40mL/min;

scanning procedure: heating from room temperature to 205 ℃ at 10 ℃/min, and recording a heating curve;

detecting the mass of a sample: 2.14mg (using an aluminium sample tray).

The detection basis is as follows: JY/T0589.3-2020 thermal analysis method general section 3 differential scanning calorimeter detection results: as in fig. 2.

(III) thermogravimetric analysis (TG):

detection instrument: TG209F1 thermogravimetric analyzer from NETZSCH company, germany

Detection conditions:

atmosphere: air, 20mL/min;

scanning procedure: room temperature to 700 ℃;

rate of temperature rise: 10 ℃/min.

The detection basis is as follows: thermal analysis methods general section 4: thermogravimetric JY/T0589.4-2020

Detection result: as in fig. 3.

(IV) Infrared Spectroscopy (IR):

detection instrument: FT-IR NICOLET 6700 (U.S.A.)

Detection conditions: potassium bromide tabletting method

The detection basis is as follows: general rules of GB/T6040-2019 infrared spectrum analysis method

Detection result: as in fig. 4.

(fifth) High Performance Liquid Chromatography (HPLC):

detection instrument: agilent 1260 Infinicity II high performance liquid chromatograph (U.S.)

Detection conditions:

chromatographic column: porosill 120EC-C18

Mobile phase: water/acetonitrile (80:20)

Column temperature: 30 ℃; detection wavelength: 254nm.

The detection basis is as follows: high performance liquid chromatography (VD) of second appendix of Chinese pharmacopoeia

Detection result: as in fig. 5.

Example 2

100g of crude 2- (4- (7- (2-fluoro-4- (1- ((4-fluorophenyl) carbamoyl) cyclopropane-1-carboxamide) phenoxy) thieno [3,2-b ] pyridin-2-yl) -1H-pyrazolyl-1-yl) ethyl valine ester hydrochloride are weighed, added to a reaction flask, 800mL of a mixed solution of methanol and water (V methanol: V water=9:1) are added, and the mixture is heated to reflux to 70℃with stirring. Stirring for 30min after dissolution, cooling to 15-25 ℃, stirring for crystallization for 12h, filtering, and drying a filter cake to obtain 75g of solid; adding the obtained solid into a reaction bottle, adding 150ml of methanol, pulping for 4 hours at 20-25 ℃, and carrying out suction filtration; the filter cake obtained was air-dried at 50℃to give 71g of white powder, with a total yield of 71% in both steps. The resulting compound is crystalline form a of 2- (4- (7- (2-fluoro-4- (1- ((4-fluorophenyl) carbamoyl) cyclopropane-1-carboxamide) phenoxy) thieno [3,2-b ] pyridin-2-yl) -1H-pyrazolyl-1-yl) ethyl valine ester hydrochloride.

Example 3

100g of crude 2- (4- (7- (2-fluoro-4- (1- ((4-fluorophenyl) carbamoyl) cyclopropane-1-carboxamide) phenoxy) thieno [3,2-b ] pyridin-2-yl) -1H-pyrazolyl-1-yl) ethyl valine ester hydrochloride are weighed, added to a reaction flask, 1500ml of tetrahydrofuran solution is added, and the mixture is heated to reflux under stirring to 80 ℃. Stirring for 30min after dissolution, cooling to 15-25 ℃, stirring for crystallization for 12h, filtering, and drying a filter cake to obtain 85g of solid; adding the obtained filter cake into a reaction bottle, adding 170ml of methanol, pulping for 4 hours at 20-25 ℃, and carrying out suction filtration; the filter cake obtained was air-dried at 50℃to give a white powder 81, with a total yield of 81% in both steps. The resulting compound is crystalline form a of 2- (4- (7- (2-fluoro-4- (1- ((4-fluorophenyl) carbamoyl) cyclopropane-1-carboxamide) phenoxy) thieno [3,2-b ] pyridin-2-yl) -1H-pyrazolyl-1-yl) ethyl valine ester hydrochloride.

Example 4

50g of crude 2- (4- (7- (2-fluoro-4- (1- ((4-fluorophenyl) carbamoyl) cyclopropane-1-carboxamide) phenoxy) thieno [3,2-b ] pyridin-2-yl) -1H-pyrazolyl-1-yl) ethyl valine ester hydrochloride was weighed separately and added to a reaction flask, and the compound A of the present invention was prepared according to the experimental procedure of example 1 using the experimental conditions of Table 2, and the total yield is shown in Table 2.

TABLE 2

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Conclusion of experiment: the type of mixed solvent used in the experiment is preferably tetrahydrofuran/water; the alcohol solvent is preferably methanol; the ketone solvent used is preferably acetone. The volume ratio (V/V) of the mixed solvent to the water is 3:1-15:1, preferably 9:1; the ratio of the crude product to the solvent is calculated according to g/mL, and the weight volume ratio is 1: 4-20, tetrahydrofuran is preferably 1:10, and methanol is preferably 1:8. The solution is preferably heated to 50-90 ℃, more preferably tetrahydrofuran to 80 ℃, and methanol to 70 ℃; according to this embodiment, the solid is allowed to stand for 2 to 16 hours, more preferably 12 hours. The precipitation temperature is 0 to 30 ℃, preferably 15 to 25 ℃. Filtering after solid separation is completed, adding methanol for pulping, and performing crystal transformation operation; the weight-volume ratio of the solid obtained by recrystallization to the methanol is 1:1 to 10, preferably 1:2; the pulping temperature is 10-50 ℃, preferably 20-30 ℃; the beating time is 1 to 10 hours, more preferably 4 hours. After the crystal transformation is finished, the obtained suspension is filtered to obtain white powder, and the white powder is dried, wherein the drying temperature is 30-70 ℃, preferably 50 ℃.

Example 5 stability test

Stability study of form a of 2- (4- (7- (2-fluoro-4- (1- ((4-fluorophenyl) carbamoyl) cyclopropane-1-carboxamide) phenoxy) thieno [3,2-b ] pyridin-2-yl) -1H-pyrazolyl-1-yl) ethyl valine ester hydrochloride.

The stability of the obtained A crystal form is investigated (10 days of acceleration test), and the purity, the maximum single impurity and the total impurity of the A crystal form are compared with data of 0 days under the conditions of high temperature 60 ℃ and high humidity 92.5 percent and illumination. The experimental result shows that the purity is slightly reduced under the illumination condition, and the obtained crystal form is stable under other conditions. The A crystal form has higher stability under the high-temperature and high-humidity conditions.

TABLE 3 results of Crystal form influence factor test

Crude 2- (4- (7- (2-fluoro-4- (1- ((4-fluorophenyl) carbamoyl) cyclopropane-1-carboxamide) phenoxy) thieno [3,2-b ] pyridin-2-yl) -1H-pyrazolyl-1-yl) ethyl valine ester hydrochloride was subjected to stability investigation (10 day acceleration test) and the data of the purity, maximum single impurity and total impurity of the crude product were compared at high temperature 40, 60 ℃ for 0 days. Experimental results show that the crude product is stored for 5 days and 10 days at the temperature of 60 ℃, the purity is reduced more, and the stability is lower than that of the A crystal form stored under the same condition.

TABLE 4 influence factor test results of crude product

EXAMPLE 6 solubility test

The prepared crystal form a and crude product of example 1 were subjected to solubility test in water, and the solubility test method was referred to the test method in the second part of the pharmacopoeia of 2020 edition.

The test method comprises the following steps: weighing a test substance ground into fine powder or measuring a liquid test substance, and strongly shaking for 30 seconds every 5 minutes in a solvent with a certain capacity at 25+/-2 ℃; dissolution within 30 minutes, such as when no visually observable solute particles or droplets are observed, is considered complete dissolution.

The approximate solubility of a pharmaceutical product is expressed in terms of the following terms:

is very soluble	Means that 1g of solute can be dissolved in less than 1ml of solvent
		Is easily dissolved	1g of solute can be dissolved in 1-10 ml of solvent
Dissolving	1g of solute can be dissolved in 10-30 ml of solvent
		Slightly soluble	1g of solute can be dissolved in 30-100 ml of solvent
Slightly soluble	1g of solute can be dissolved in 100-1000 ml of solvent
		Very slightly dissolving	1g of solute can be dissolved in 1000-10 000ml of solvent
Hardly or not soluble	Means that 1g of solute cannot be completely dissolved in 10000ml of solvent

The results of the solubility test of form a and crude in water are shown in table 5:

TABLE 5 solubility test results of form A and crude in water

Experimental results show that the solubility of the crystal form A of the compound obtained by the invention in water is obviously better than that of a crude product of the compound, and various index requirements of preparation development can be met.

Example 7 bioavailability test

Experimental materials: SD male mice, SPF grade. Shanghai Sipuler-BiKai laboratory animal Co., ltd. Other reagents are all commercially available products.

(1) Solution preparation

Weighing a proper amount of the sample to be tested into a glass bottle, adding PEG400, carrying out vortex oscillation, uniformly mixing, and carrying out ultrasonic treatment to obtain a clear solution. The test article was administered to the experimental animal (n=3) by gavage at a dose of 200 mg/kg.

(2) Test method

SD rats were weighed prior to dosing and dosing was calculated based on body weight. The medicine is orally taken through stomach irrigation. Blood was collected via submaxillary vein or other means at 0,0.25,0.5,1,2,4,8,12,24 hour intervals, each sample was collected at approximately 0.20ml, edta-K2 was anticoagulated, and placed on ice after collection. And the plasma was centrifuged within 1 hour (centrifugation conditions: 6800g,6min,2-8 ℃). The plasma samples were stored at-40 to-20 ℃ for analysis. The compounds of the present invention were all converted to td32-4 after 4 hours of intravenous administration and the data were taken as 100% bioavailability and used as calculation of oral bioavailability.

(3) Experimental results

Form a prepared in example 1: can be rapidly converted into td32-4 in vivo when 200mg/kg is administered, and the oral bioavailability f=54.3% is calculated according to the exposure of the converted td32-4

Crude product of example 1: can be rapidly converted into td32-4 in vivo when 200mg/kg is administered, and the oral bioavailability f=32.6% is calculated according to the exposure of the converted td32-4

From the pharmacokinetic data of crystalline form a of 2- (4- (7- (2-fluoro-4- (1- ((4-fluorophenyl) carbamoyl) cyclopropane-1-carboxamide) phenoxy) thieno [3,2-b ] pyridin-2-yl) -1H-pyrazolyl-1-yl) ethyl valine ester hydrochloride with the crude product, it can be seen that crystalline form a of the compound of the present invention exhibits significantly better pharmacokinetic properties in vivo than the crude product, and that the bioavailability of crystalline form a is significantly higher at the 200mg/kg dose than the crude product at the same dose.

Example 8 influence of pressure/grinding on crystalline form

100mg of form A of example 1 was weighed and compressed directly into tablets (7.1 kN,8.5 kN) using a tablet press at a pressure well above the main pressure at the time of tabletting. The XPRD is measured after the obtained medicine is ground, and the result shows that the characteristic diffraction peak at the 2 theta degree position is not changed significantly, which indicates that the crystal form is stable in the tabletting and grinding processes and meets the preparation requirement.

EXAMPLE 9 preparation of solid pharmaceutical formulations

Prescription 1: tablet formulation containing 5% of form A prepared in example 1

The preparation method comprises the following steps: the above components are mixed according to a conventional preparation method and directly compressed into tablets.

Prescription 2: tablet formulation containing 5% of crude product of example 1

The preparation method comprises the following steps: the above components are mixed according to a conventional preparation method and directly compressed into tablets.

EXAMPLE 10 solid pharmaceutical formulation Performance test

The solubility, compressibility and disintegration of formulations 1 and 2 were tested according to the 2010 pharmacopoeia dissolution test. The result shows that under the condition of consistent auxiliary materials, the dissolution rate of the prescription 1 reaches more than 80% in 15 minutes, and the dissolution rate of the prescription 2 is lower than 80%; and the compressibility and the disintegration degree of the prescription 1 are both superior to those of the prescription 2, so that the crystal form A can meet various index requirements of the preparation better than the crude product.

The above description of the preferred embodiment of the present invention is not intended to limit the present invention, and those skilled in the art can make various changes or modifications according to the present invention without departing from the spirit of the present invention, and shall fall within the scope of the appended claims.

Claims (10)

1. A crystalline form A of the compound 2- (4- (7- (2-fluoro-4- (1- ((4-fluorophenyl) carbamoyl) cyclopropane-1-carboxamide) phenoxy) thieno [3,2-b ] pyridin-2-yl) -1H-pyrazolyl-1-yl) ethyl valine ester hydrochloride, the structural formula of the compound is shown as formula (I),

wherein when Cu-K alpha radiation is used, the X-ray powder diffraction pattern data of the compound crystal form A are as follows:

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2. form a of compound according to claim 1, wherein the thermogravimetric analysis profile of form a of compound shows a loss of weight of 2.55% in the range of from room temperature to 136.0 ℃; a loss of weight in the range of 136.0 ℃ to 204.0 ℃ of 1.32%; a loss of weight in the range 204.0 ℃ to 481.0 ℃ of 53.58%; the loss weight is 42.31% in the range of 481.0 ℃ to 700.0 ℃.

3. Form a compound according to claim 1, characterized by a differential scanning calorimetry spectrum of form a in which there are two endotherms at 111.1 ℃ and 197.4 ℃.

4. Form a of compound according to claim 1 or 2, characterized by an infrared absorption spectrum of form a of said compound measured by compression of potassium bromide at 3356cm ^-1 、3070cm ^-1 、3027cm ^-1 、2963cm ^-1 、2879cm ^-1 、2785cm ^-1 、2683cm ^-1 、2618cm ^-1 、1752cm ^-1 、1693cm ^-1 、1642cm ^-1 、1602cm ^-1 、1556cm ^-1 、1507cm ^-1 、1495cm ^-1 、1469cm ^-1 、1407cm ^-1 、1327cm ^-1 、1288cm ^-1 、1251cm ^-1 、1200cm ^-1 、868cm ^-1 、829cm ^-1 Has an absorption peak.

5. A process for the preparation of form a of the compound of claim 1, comprising the steps of:

1) Adding a crude product of 2- (4- (7- (2-fluoro-4- (1- ((4-fluorophenyl) carbamoyl) cyclopropane-1-carboxamide) phenoxy) thieno [3,2-b ] pyridin-2-yl) -1H-pyrazolyl-1-yl) ethyl valine ester hydrochloride into a solvent, and heating and refluxing until the crude product is dissolved to obtain a compound solution;

the solvent is one selected from tetrahydrofuran, ethyl acetate, C1-C4 alkyl alcohol and C3-C4 alkyl ketone, or a mixed solvent of the organic solvent and water;

in the mixed solvent, the volume ratio of the organic solvent to the water is 3:1-15:1;

the weight-volume ratio of the crude compound to the solvent is 1g (4-20) mL;

the heating temperature is 50-90 ℃;

2) After the compound solution is clarified, cooling the compound solution until solid is separated out;

the solid separation time is 2-16 hours, and the solid separation temperature is 0-30 ℃;

3) Filtering and collecting the solid, and subsequently drying the solid;

4) Adding methanol into the solid, pulping to enable the solid to be crystallized in the methanol, and obtaining suspension;

the weight volume ratio of the solid to the beating methanol is 1g (1-10) mL; the crystal transformation temperature is 10-50 ℃; the crystal transformation time is 1-10 hours;

5) Filtering the suspension obtained in the step 4) to obtain white powder, and drying by blowing to obtain the compound crystal form A; the drying temperature is 30-70 ℃.

6. The process according to claim 5, wherein the C1-C4 alkyl alcohol is one selected from the group consisting of methanol, ethanol, propanol and isopropanol; the C3-C4 alkyl ketone is selected from one of acetone and methyl ethyl ketone.

7. The process according to claim 5, wherein the C1-C4 alkyl alcohol in step 1) is methanol; the C3-C4 alkyl ketone is acetone.

8. The preparation method according to claim 5, wherein the solvent in the step 1) is a mixed solvent of tetrahydrofuran and water in a volume ratio of 9:1; the weight volume ratio of the crude compound to the mixed solvent is 1g:10mL, heating temperature is 80 ℃;

the solid separating time in the step 2) is 12 hours; the solid separation temperature is 15-25 ℃;

the weight volume ratio of the crystal-transferred solid to the beating methanol is 1g:2mL; the crystal transformation temperature is 20-30 ℃; the crystal transformation time is 4 hours;

the drying temperature was 50 ℃.

9. The preparation method according to claim 5, wherein the solvent in step 1) is a mixed solvent of methanol and water in a volume ratio of 9:1; the weight volume ratio of the crude compound to the mixed solvent is 1g:8mL, heating temperature is 70 ℃;

the solid separating time in the step 2) is 12 hours; the solid separation temperature is 15-25 ℃;

the weight volume ratio of the crystal-transferred solid to the beating methanol is 1g:2mL; the crystal transformation temperature is 20-30 ℃; the crystal transformation time is 4 hours;

the drying temperature was 50 ℃.

10. A pharmaceutical composition comprising form a of the compound of claim 1.

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