CN111349076B

CN111349076B - Crystal form of L-lysine salt of benzopiperidine derivative and preparation method thereof

Info

Publication number: CN111349076B
Application number: CN201911316928.XA
Authority: CN
Inventors: 邵成; 尤凌峰; 冯君; 贺峰
Original assignee: Jiangsu Hengrui Medicine Co Ltd; Shanghai Hengrui Pharmaceutical Co Ltd
Current assignee: Jiangsu Hengrui Medicine Co Ltd; Shanghai Hengrui Pharmaceutical Co Ltd
Priority date: 2018-12-20
Filing date: 2019-12-19
Publication date: 2022-07-26
Anticipated expiration: 2039-12-19
Also published as: CN111349076A

Abstract

The disclosure provides a crystal form of a benzopiperidine derivative L-lysine salt and a preparation method thereof. Specifically, the disclosure provides a crystal form II of L-lysine of (E) -3- (4- ((1R,3R) -2- (4-cyclopropylphenyl) -6- (1-ethyl-1H-pyrazol-4-yl) -3-methyl-1, 2,3, 4-tetrahydroisoquinolin-1-yl) phenyl) acrylic acid and a preparation method thereof. The II crystal form of the compound disclosed in the formula (I) has good stability and can be better used for clinical treatment.

Description

Crystal form of L-lysine salt of benzopiperidine derivative and preparation method thereof

Technical Field

The present disclosure relates to a crystal form II of (E) -3- (4- ((1R,3R) -2- (4-cyclopropylphenyl) -6- (1-ethyl-1H-pyrazol-4-yl) -3-methyl-1, 2,3, 4-tetrahydroisoquinolin-1-yl) phenyl) acrylic acid L-lysine salt, a preparation method thereof, and applications of the crystal form II and a composition in preparing medicines for preventing and/or treating diseases or symptoms mediated or dependent on estrogen receptors.

Background

Estrogens are steroid hormones secreted by the endocrine system and play important roles in the reproductive system, bone tissue, cardiovascular system, immune system and central nervous system. The estrogen signaling system plays an important role in regulating cell growth, differentiation and apoptosis. The occurrence and development of estrogen-dependent tumors such as breast cancer, ovarian cancer, endometrial cancer, osteoporosis, schizophrenia, senile dementia and the like are closely related to estrogen.

An important protein of the estrogen signaling pathway is the Estrogen Receptor (ER), which is a steroid hormone receptor, a ligand-activated transcription factor belonging to the nuclear receptor superfamily, and comprises two subtypes: ER α (found in 1950) and ER β (found in 1996), each encoded by a different gene. ER α is distributed primarily in the uterus, ovary, testis, pituitary, kidney, epididymis, and adrenal gland, while ER β is distributed primarily in the prostate, ovary, lung, bladder, brain, and blood vessels. Since either full agonists or full antagonists have more serious side effects, the study of SERMs has been ongoing. By "selective" is meant that the SERM behaves as an agonist in certain tissues such as bone, liver, ER β focal region of the cardiovascular system, and as an antagonist in other tissues such as breast. It may be an agonist or antagonist in the uterus (more prominent region of ER α). Tamoxifen, which belongs to a class of compounds called Selective Estrogen Receptor Modulators (SERMs), has the effect of stabilizing era and slightly upregulating era receptor levels; in contrast, fulvestrant (fulvestrant) causes rapid degradation of ER α, exacerbating blockade of the ER receptor signalling pathway, and such compounds are known as selective estrogen receptor down-regulators (SERDs). These differences in the mechanism of action of SERMs and SERDs also appear to be the mechanism responsible for the resistance of these compounds. Many tumors that are tamoxifen resistant while the ER remains positive are sensitive to fulvestrant. It has been found clinically that SERDs such as fulvestrant are effective in treating some ER α positive, tamoxifen resistant breast cancers. Thus, compounds that cause ER-alpha degradation can be used to extend the duration of efficacy in breast cancer patients successfully treated with antiestrogen therapy (possibly using different SERMs, aromatase inhibitors and SERDs in sequence).

Published selective estrogen receptor-mediated modulator patent applications include WO2014165723, WO2014151899, WO2014141292, WO2014191726, WO2015092634, WO2014135834, WO2014106848, and EP 1113007A.

WO2017107754 discloses a high-efficiency and low-toxicity SERD compound aiming at an estrogen signaling pathway, which shows good activities in the aspects of inhibition of binding of E and ER, ER degradation, MCF7 cell proliferation and the like, and particularly has more prominent advantages in the aspect of Emax value of ER degradation, and the structure of the compound is shown as formula II:

the chemical stability of the medicine is often influenced by the crystal form structure of the medicinal active ingredient, and the change of the crystal form structure of the compound is possibly caused by the difference of crystallization conditions and storage conditions, and other forms of crystal forms are sometimes generated. Generally, amorphous drug products do not have regular crystal structures and often have other defects, such as poor product stability, fine crystallization, difficult filtration, easy agglomeration, poor flowability and the like. Therefore, there is a need to improve various properties of the above products.

Disclosure of Invention

The present disclosure provides a crystal form II of a compound shown in a formula (I), an X-ray powder diffraction pattern expressed by a diffraction angle 2 theta angle is provided, the diffraction angle 2 theta angle of the crystal form II has characteristic peaks at 7.52, 10.38, 12.64, 18.70, 20.22, 20.90, 21.96 and 23.08,

in some embodiments, the crystalline form II, having an X-ray powder diffraction pattern expressed in diffraction angle 2 Θ angles, has characteristic peaks at 7.52, 10.38, 12.64, 17.02, 18.70, 20.22, 20.90, 21.96, 23.08, 24.50, and 27.10.

In some embodiments, the crystalline form II, having an X-ray powder diffraction pattern expressed in degrees 2 Θ at diffraction angle, has characteristic peaks at 6.92, 7.52, 8.16, 10.08, 10.38, 12.64, 15.28, 15.60, 16.01, 17.02, 18.08, 18.70, 20.22, 20.90, 21.96, 22.76, 23.08, 23.56, 24.50, 26.36, 27.10, 28.64, 30.54, 31.50, 34.24, 38.84, and 39.62.

In other embodiments, the crystalline form II has an X-ray powder diffraction pattern expressed in terms of diffraction angle, 2 Θ, as shown in fig. 1.

In an alternative embodiment, the obtained compound represented by formula (II) (i.e., the crystal form II of (E) -3- (4- ((1R,3R) -2- (4-cyclopropylphenyl) -6- (1-ethyl-1H-pyrazol-4-yl) -3-methyl-1, 2,3, 4-tetrahydroisoquinolin-1-yl) phenyl) acrylic acid) has significantly improved stability compared with the free form (the compound represented by formula (I)), and has significant advantages in terms of purity, long-term stability, hygroscopicity, and the like, and better drug developability.

The disclosure also provides a method for preparing a crystal form II of the compound shown in the formula (I).

A first process for preparing a crystalline form of compound II of formula (I), comprising:

adding a compound shown as a formula (II) into a solvent (I), stirring for dissolving or heating for dissolving, adding L-lysine or a solution/suspension of the L-lysine and the solvent (I), and stirring for crystallizing, wherein the solvent (I) is at least one selected from an alcohol solvent, an ester solvent or a mixed solvent of the alcohol solvent and the ester solvent and water, preferably at least one selected from methanol, ethanol, isopropanol, ethyl acetate or a mixed solvent of the alcohol solvent, the ester solvent and water,

in some embodiments, the percentage of water in solvent (I) in the present method is 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%.

In some embodiments, the volume (ml) of solvent (I) used in the present process is 1 to 100 times the weight (g) of the compound, and may be 1,2,3,4, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100 times.

A second process for preparing a crystalline form II of compound of formula (I), comprising:

adding the compound shown in the formula (I) into a solvent (II), stirring for dissolving or heating for dissolving, and then stirring for crystallization, wherein the crystallization mode is selected from room temperature crystallization, cooling crystallization, solvent volatilization crystallization or crystal seed addition induced crystallization, and preferably is selected from crystal seed addition induced crystallization; the solvent (II) is at least one selected from alcohol solvents or mixed solvents of the alcohol solvents and water, preferably at least one selected from methanol, ethanol or isopropanol or mixed solvents of the alcohol solvents and water;

in some embodiments, the volume (ml) of solvent (II) used in the present process is 1 to 100 times the weight (g) of the compound, and may be 1,2,3,4, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100 times.

In some embodiments, the percentage of water in solvent (II) in the present methods is 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%.

A third process for preparing a crystalline form of compound II of formula (I), comprising:

adding the compound shown in the formula (I) into a solvent (III), and pulping, wherein the solvent (III) is selected from a mixed solvent of a ketone solvent and a nitrile solvent with water, and preferably acetone/water and acetonitrile/water.

In some embodiments, the volume (ml) of solvent (III) used in the present process is 1 to 60 times the weight (g) of the compound, and may be 1,2,3,4, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60 times.

In some embodiments, the method for preparing the crystalline form II in the present disclosure further includes the steps of filtering, washing, drying, and the like, wherein the drying mode may be normal pressure drying or reduced pressure drying, preferably reduced pressure drying; the drying temperature is 25-60 deg.C, 25, 30, 35, 40, 45, 50, 55, 60 deg.C, preferably 30-50 deg.C.

In an alternative embodiment, drying in the present disclosure is carried out under reduced pressure (relative vacuum may be in the range of-0.080 to-0.098 MPa).

The cooling crystallization in the disclosure is a process of slowly or rapidly cooling a solution to be crystallized to a temperature of-10 ℃ to 25 ℃ and stirring for crystallization.

In the present disclosure, "solvent volatilization crystallization" refers to a process in which a solvent in a solution to be crystallized gradually volatilizes and crystallizes.

The term "crystallization at room temperature" in the present disclosure means a process of slowly cooling a solution to be crystallized to room temperature and stirring for crystallization.

The present disclosure provides a pharmaceutical composition, which is prepared from the compound II crystal form shown in the formula (I) and one or more pharmaceutically acceptable carriers, diluents or excipients. For example, the crystal form II of the compound represented by formula (I) or the pharmaceutical preparation of the present disclosure may be formulated as tablets, capsules, pills, granules, solutions, suspensions, syrups, injections (including injections, sterile powders for injection and concentrated solutions for injection), suppositories, inhalants or sprays.

In addition, the pharmaceutical compositions of the present disclosure may also be administered to a patient or subject in need of such treatment by any suitable mode of administration, e.g., oral, parenteral, rectal, pulmonary, or topical administration. When used for oral administration, the pharmaceutical composition can be prepared into oral preparations, such as oral solid preparations, such as tablets, capsules, pills, granules and the like; or oral liquid preparations such as oral solution, oral suspension, syrup, etc. When formulated into oral preparations, the pharmaceutical preparations may further comprise suitable fillers, binders, disintegrating agents, lubricants and the like. When used for parenteral administration, the pharmaceutical preparation can be prepared into injections, including injection solutions, sterile powders for injection, and concentrated solutions for injection. When prepared into injections, the pharmaceutical composition may be manufactured by a conventional method in the existing pharmaceutical field. When preparing injection, the pharmaceutical preparation can be added with no additive, or added with proper additive according to the nature of the medicine. When used for rectal administration, the pharmaceutical preparation may be formulated into suppositories and the like. For pulmonary administration, the pharmaceutical formulation may be formulated as an inhalant or spray. In certain preferred embodiments, the crystalline form II of the compound of formula (I) of the present disclosure is present in a therapeutically and/or prophylactically effective amount in a pharmaceutical composition or medicament. In certain preferred embodiments, the crystalline form II of the compound of formula (I) of the present disclosure is present in a pharmaceutical composition or medicament in the form of a unit dose.

The present disclosure provides a use of crystalline form II, pharmaceutical compositions containing crystalline form II in the manufacture of a medicament for treating an estrogen receptor mediated or dependent disease or disorder. Further, the present application provides a method of inhibiting a disease associated with an estrogen receptor comprising administering to a subject in need thereof a therapeutically and/or prophylactically effective amount of the crystalline form II of the compound of formula (I) of the present disclosure, or a pharmaceutical composition of the present disclosure.

The estrogen receptor-mediated or dependent diseases or conditions described in this disclosure are selected from cancers, central nervous system defects, cardiovascular system defects, blood system defects, immune and inflammatory diseases, susceptible infections, metabolic defects, neurological defects, psychiatric defects, or reproductive defects; the cancer is selected from breast cancer, endometrial cancer, cervical cancer, skin cancer, prostate cancer, ovarian cancer, fallopian tube tumors, ovarian tumors, hemophilia or leukemia, preferably breast cancer, ovarian cancer, endometrial cancer, prostate cancer or uterine cancer, more preferably breast cancer.

According to the guiding principle of moisture-attracting property of 9103 medicament in 2015 th edition of four parts of Chinese pharmacopoeia and the definition of moisture-attracting weight increment,

deliquescing: absorbing sufficient water to form a liquid;

has the characteristics of moisture absorption: the moisture-inducing weight is not less than 15%;

has the following moisture absorption: the moisture-inducing weight gain is less than 15% but not less than 2%;

slightly hygroscopic: the moisture-inducing weight is less than 2% but not less than 0.2%;

no or little hygroscopicity: the moisture-drawing weight gain is less than 0.2 percent.

The crystal form II of the compound shown in the formula (I) in the disclosure has moisture absorption weight increased by 1.819% and is slightly moisture-absorbing under the condition of 10.0% RH-80.0% RH.

And performing structure determination and crystal form research on the obtained II crystal form of the compound shown in the formula (I) through X-ray powder diffraction pattern (XRPD) and Differential Scanning Calorimetry (DSC).

The "X-ray powder diffraction pattern" described in this disclosure is measured using Cu-ka radiation.

The differential scanning calorimetry or DSC in the present disclosure refers to measuring the temperature difference and heat flow difference between a sample and a reference substance during the temperature rise or constant temperature process of the sample to characterize all the physical changes and chemical changes related to the thermal effect and obtain the phase change information of the sample.

The method for recrystallizing form II is not particularly limited, and can be carried out by a conventional recrystallization procedure. For example, the desired crystals can be obtained by dissolving the compound represented by the formula (I) as a raw material in an organic solvent, adding an anti-solvent to the solution, crystallizing the solution, and then filtering and drying the crystals.

The term "X-ray powder diffraction pattern or XRPD" as used in this disclosure refers to the pattern of X-rays according to bragg formula 2d sin θ ═ n λ (where λ is the wavelength of the X-rays,

the order n of diffraction is any positive integer, a first-order diffraction peak is generally taken, n is 1, when X-rays are incident on an atomic plane with a d-lattice plane spacing of a crystal or a part of a crystal sample at a grazing angle theta (complementary angle of incidence, also called Bragg angle), the Bragg equation can be satisfied, and the set of X-ray powder diffraction patterns can be measured.

The "2 theta or 2 theta angle" referred to in this disclosure refers to the diffraction angle, theta is the bragg angle, and is in degrees or degrees; the error range of each characteristic peak 2 theta is + -0.20, and may be-0.20, -0.19, -0.18, -0.17, -0.16, -0.15, -0.14, -0.13, -0.12, -0.11, -0.10, -0.09, -0.08, -0.07, -0.06, -0.05, -0.04, -0.03, -0.02, -0.01, 0.00, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.10, 0.11, 0.12, 0.13, 0.14, 0.15, 0.16, 0.17, 0.18, 0.19, 0.20.

The term "interplanar spacing or interplanar spacing (d value)" as used in this disclosure means that the spatial lattice selects 3 non-parallel unit vectors a, b, c connecting two adjacent lattice points, which divide the lattice into juxtaposed parallelepiped units, called interplanar spacing. The space lattice is divided according to the determined connecting lines of the parallelepiped units to obtain a set of linear grids called space grids or lattices. The lattice and the lattice are different crystal planes whose interplanar spacing (i.e., two adjacent planes) reflects the periodicity of the crystal structure by geometrical points and lines, respectivelyThe distances between the row facets) are different; has the unit of

Or angstroms.

The crystallization method disclosed by the disclosure comprises room-temperature crystallization, cooling crystallization, volatilization crystallization, seed crystal addition induced crystallization and the like.

The starting materials used in the preparation method of the crystal forms disclosed in the present disclosure may be any form of the compound represented by formula (I), and specific forms include, but are not limited to: amorphous, random crystalline, etc.

The disclosure states as "C _1-6 Alkyl "denotes a straight or branched chain alkyl group having 1 to 6 carbon atoms, and specific examples include, but are not limited to: methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, 2-methylbutyl, neopentyl, 1-ethylpropyl, n-hexyl, isohexyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 3-dimethylbutyl, 2-dimethylbutyl, 1-dimethylbutyl, 1, 2-dimethylbutyl, 1, 3-dimethylbutyl, 2-ethylbutyl, 1, 2-dimethylpropyl, and the like.

As used in this disclosure, "alcoholic solvent" means one or more "hydroxy" substituted "C _1-6 A radical derived from one or more hydrogen atoms of an alkyl radical, said radical "C _1-6 Alkyl "is as defined above, specific examples include, but are not limited to: methanol, ethanol, isopropanol, n-propanol, isoamyl alcohol or trifluoroethanol.

The "ester solvent" described in the present disclosure refers to a combination of a lower organic acid having 1 to 4 carbon atoms and a lower alcohol having 1 to 6 carbon atoms, and specific examples include, but are not limited to: ethyl acetate, isopropyl acetate or butyl acetate.

The "ketone solvent" as used in the present disclosure refers to a compound in which a carbonyl group (-c (o)) is bonded to two hydrocarbon groups, and the ketone can be classified into aliphatic ketone, alicyclic ketone, aromatic ketone, saturated ketone and unsaturated ketone according to the difference of the hydrocarbon groups in the molecule, and specific examples include, but are not limited to: acetone, acetophenone, methyl isobutyl ketone or methyl pyrrolidone.

As used in this disclosure, "nitrile solvent" means one or more "cyano" groups substituted for "C _1-6 A radical derived from one or more hydrogen atoms of alkyl, said "C _1-6 Alkyl "is as defined above, specific examples include, but are not limited to: acetonitrile or propionitrile.

The "mixed solvent" in the disclosure refers to a solvent formed by mixing one or more different organic solvents according to a certain proportion, or a solvent formed by mixing an organic solvent and water according to a certain proportion; the mixed solvent is preferably a mixed solvent of alcohols and water, or a mixed solvent of ketones and water; the alcohols and ketones are as defined above.

The instruments/conditions used in the experiments in this disclosure:

x-ray powder diffraction detection: the Rigaku UltimaIV model combined multifunctional X-ray diffractometer specifically acquires information: cu anode (40kV, 40mA), Cu-Ka 1 ray

Scanning rate 20 °/min, scanning range (2q range): 3-45 degrees, scanning step length 0.02 and slit width 0.01.

Differential scanning calorimetry: TA Q2000, temperature rising rate of 10 ℃/min, temperature of 30-300 ℃ and nitrogen purging speed of 50 mL/min.

Thermogravimetric analysis: TAQ500, the heating rate is 10 ℃/min, the specific temperature range refers to a corresponding graph, and the nitrogen purging speed is 60 mL/min.

Dynamic moisture adsorption: TAQ5000VSA, humidity is 10-90% at 25 deg.C, step by 10%, the quality change is less than 0.01% within 10000min, and the process is repeated for two circles.

Drawings

FIG. 1: an XRPD pattern of form II of the compound of formula (I).

FIG. 2: DSC of the II crystal form of the compound of the formula (I).

FIG. 3: TGA profile of the crystalline form II of the compound of formula (I).

FIG. 4 is a schematic view of: a DVS profile of form II of the compound of formula (I).

FIG. 5: XRPD comparison spectrums before and after a crystal form II hygroscopicity experiment of the compound of the formula (I) (a is an XRPD pattern after the hygroscopicity experiment, and b is an XRPD pattern before the hygroscopicity experiment).

FIG. 6: XRPD pattern of form II of compound of formula (I) after standing for 3 months at 25 ℃, 60% RH.

FIG. 7: XRPD pattern of form II of compound of formula (I) after standing for 3 months at 40 ℃, 75% RH.

FIG. 8: XRPD pattern of I crystal form of compound of formula (I)

FIG. 9: XRPD pattern of compound of formula (I) (product of comparative example 2).

FIG. 10: XRPD pattern of compound of formula (II) (product of comparative example 1)

Detailed Description

The present disclosure will be explained in more detail with reference to examples, which are only used for illustrating the technical solutions of the present disclosure and do not limit the spirit and scope of the present disclosure.

Comparative example 1 preparation of Compound of formula (II) (method in WO 2017107754)

First step of

3- (2-Nitropropyl-1-en-1-yl) phenol 4b

M-hydroxybenzaldehyde 4a (10g, 81.9mmol), nitroethane (60g, 819mmol) and ammonium acetate (1.54g, 20mmol) were added to a reaction flask, heated to 80 ℃, methylamine (1g, 32.2mmol) was added, and after the addition, the reaction was stirred for 2 hours. Water (50mL) was added to the reaction solution, extracted with ethyl acetate (30mL × 3), the organic phases were combined, dried over anhydrous sodium sulfate, the drying agent was removed by filtration, the filtrate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography with n-hexane and ethyl acetate as eluents to give the title product 4b (9.5g, yellow solid), yield: and (4) 64.6%.

Second step of

1- (3-hydroxyphenyl) propan-2-one 4c

4b (9.5g, 53mmol) was added to a mixed solution of methanol and water (V: V ═ 10:1, 110mL), raney nickel (10%, 9.5g) and acetic acid (3.2g, 53mmol) were added, and after the addition, hydrogen gas substitution was carried out three times, and the reaction was stirred for 16 hours. Filtration, evaporation of most of the solvent from the filtrate, extraction with ethyl acetate (50mL × 3), combination of the organic phases, drying over anhydrous sodium sulfate, removal of the drying agent by filtration, concentration of the filtrate under reduced pressure, and purification of the resulting residue by silica gel column chromatography with n-hexane and ethyl acetate as eluents gave the title product 4c (3.7g, yellow oil), yield: 46.8 percent.

The third step

3- (2- ((4-cyclopropylphenyl) amino) propyl) phenol 4e

4-Cyclopropylaniline hydrochloride 4d (390mg, 2.30mmol, Do you Zi) was dissolved in dichloroethane (10mL), triethylamine (233mg, 2.30mmol) was added thereto, and the mixture was stirred for 5 minutes, 4c (345mg, 2.30mmol) and sodium triacetoxyborohydride (730mg, 3.45mmol) were added thereto, and the reaction was stirred for 12 hours. The reaction was added with water (10mL), extracted with dichloromethane (10mL × 3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered to remove the drying agent, the filtrate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography with n-hexane and ethyl acetate as eluents to give the title product 4e (540mg, brown dope), yield: 87.8 percent.

The fourth step

(E) -methyl 3- (4- (2- (4-cyclopropylphenyl) -6-hydroxy-3-methyl-1, 2,3, 4-tetrahydroisoquinolin-1-yl) phenyl) acrylate 10a

4e (540mg, 2.02mmol), 1e (576mg, 3.03mmol) and triisopropylchlorosilane (1.95g, 10.10mmol) were added to N, N-dimethylformamide (10mL), and after the addition, the mixture was heated to 120 ℃ and stirred for reaction for 3 hours. The reaction solution was cooled to room temperature, concentrated under reduced pressure, and water (20mL) was added to the resulting residue, stirred uniformly, extracted with ethyl acetate (10mL × 3), the organic phases were combined, concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography with n-hexane and ethyl acetate as eluents to give the title product 10a (490mg, brown solid) in yield: 55.2 percent.

The fifth step

(E) -methyl 3- (4- ((1R,3R/1S,3S) -2- (4-cyclopropylphenyl) -3-methyl-6- (((trifluoromethyl) sulfonyl) oxy) -1,2,3, 4-tetrahydroisoquinolin-1-yl) phenyl) acrylate 10b

10a (490mg, 1.11mmol) was dissolved in dichloromethane (10mL), 2, 6-lutidine (180mg, 1.67mmol), trifluoromethanesulfonic anhydride (409mg, 1.45mmol) were added in this order under ice-bath, and the ice-bath was removed after the addition, and the reaction was stirred at room temperature for 16 hours. The reaction was quenched by the addition of water (10mL) to the reaction solution, extracted with dichloromethane (10mL × 2), the organic phases were combined, concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography with n-hexane and ethyl acetate as eluents to give the title product 10b (230mg, yellow solid), yield: 36.3 percent.

The sixth step

(E) -methyl 3- (4- ((1R,3R/1S,3S) -2- (4-cyclopropylphenyl) -6- (1-ethyl-1H-pyrazol-4-yl) -3-methyl-1, 2,3, 4-tetrahydroisoquinolin-1-yl) phenyl) acrylate 13a

10b (485mg, 0.85mmol), 1-ethyl-4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole (283mg, 1.275mmol), 1,1' -bis (diphenylphosphino) ferrocene ] dichloropalladium (63mg, 0.085mmol) was dissolved in a mixed solution of 1, 4-dioxane and water (V: V ═ 7:1, 8mL), and 2M sodium carbonate solution (0.85mL, 1.7mmol) was added, and after completion of addition, microwave reaction was carried out at 120 ℃ for 1 hour. Cooling to room temperature, addition of water (20mL), extraction with ethyl acetate (50mL × 3), combination of the organic phases, drying over anhydrous sodium sulfate, filtration, concentration of the filtrate under reduced pressure, and purification of the resulting residue by silica gel column chromatography with n-hexane and ethyl acetate as eluents gave the title product 13a (352mg, yellow solid), yield: 80 percent.

MS m/z(ESI):518.5[M+1]

Step seven

(E) -3- (4- ((1R,3R/1S,3S) -2- (4-cyclopropylphenyl) -6- (1-ethyl-1H-pyrazol-4-yl) -3-methyl-1, 2,3, 4-tetrahydroisoquinolin-1-yl) phenyl) acrylic acid 13

13a (350mg, 0.676mmol) was dissolved in a mixed solvent of methanol and tetrahydrofuran (V: V ═ 1:1, 28mL), and 2M sodium hydroxide solution (1.7mL, 3.38mmol) was added, and after the addition, the reaction was stirred for 16 hours. The reaction solution was concentrated under reduced pressure, water (10mL) was added to the resulting residue, stirred uniformly, 2N hydrochloric acid was added dropwise to the reaction solution pH to 2-3, extracted with ethyl acetate (50mL × 3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography with dichloromethane and methanol as eluents to give the title product 13(260mg, yellow solid), yield: 76 percent.

MS m/z(ESI):504.5[M+1]

The eighth step

(E) -3- (4- ((1R,3R) -2- (4-cyclopropylphenyl) -6- (1-ethyl-1H-pyrazol-4-yl) -3-methyl-1, 2,3, 4-tetrahydroisoquinolin-1-yl) phenyl) propenoic acid form (II)

Chiral preparation of 13(250mg,0.497mmol) was performed (separation conditions: chiral column Superchiral S-AD (Chiralway),2cm I.D. 25cm,5 μm, mobile phase: carbon dioxide: ethanol 60:40, flow rate: 50g/min), the corresponding fractions were collected and concentrated under reduced pressure to give the title product, formula (II) (105mg, yellow solid)

Chiral HPLC analysis: retention time 9.317 minutes, chiral purity: 100 percent. (column: Superchiral S-AD (Chiralway),0.46cm I.D.. times.15 cm,5 μm; mobile phase: carbon dioxide: ethanol ═ 60: 40).

¹ H-NMR(400MHz,DMSO-d ₆ )δ7.95(s,1H),7.79(s,1H),7.54-7.58(d,1H),7.32-7.42(m,7H),6.86-6.88(d,2H),6.75-6.77(d,2H),6.34-6.38(d,1H),5.72(s,1H),4.72(m,1H),4.16-4.22(m,2H),3.36-3.41(m,1H),2.75-2.79(d,1H),1.73-1.77(m,1H),1.45-1.49(m,3H),1.00-1.02(d,3H),0.78-0.80(m,2H),0.50-0.51(m,2H).

The XRPD pattern of the yellow solid is shown in fig. 10, with no distinct characteristic peaks.

Comparative example 2 preparation of Compound of formula (I)

The method comprises the following steps: a compound represented by the formula (II) (684.1mg, 1.4mmol) and methanol (10mL) were added to a reaction flask, L-lysine (0.435g, 2.97mmol) was added, the temperature was raised to 50 ℃ and stirring was carried out for 30 minutes, the reaction liquid was clarified, the temperature was lowered to room temperature, and the reaction liquid was concentrated under reduced pressure to give the title product (880mg, yellow solid). The X-ray powder diffraction pattern is shown in figure 9, and the crystal has no characteristic peak and is amorphous.

The method 2 comprises the following steps: adding a compound (101mg, 0.2mmol) shown in the formula (II) and 1, 4-dioxane (0.5mL) into a reaction bottle, stirring to dissolve, dispersing L-lysine (28mg, 0.19mmol) in 1, 4-dioxane (2.5mL), adding into a reaction solution, heating to 95 ℃, precipitating a solid after the reaction solution is clarified, slowly cooling to 50 ℃, stirring for 12 hours, cooling to room temperature, filtering the reaction solution, collecting a filter cake, and drying in vacuum to obtain the title product (65mg, light gray solid) with yield: 50 percent. And no characteristic peak is detected by X-ray powder diffraction, and the product is amorphous.

The method 3 comprises the following steps: adding the compound represented by the formula (II) (101mg, 0.2mmol) and toluene (0.5mL) into a reaction flask, heating to 60 ℃, stirring to dissolve, dispersing L-lysine (28mg, 0.19mmol) in toluene (1.5mL), adding into the reaction solution, stirring at 60 ℃ for 30 minutes, slowly cooling to room temperature, stirring for 16 hours, filtering the reaction solution, collecting a filter cake, and drying in vacuum to obtain the title product (70mg, brown solid) with yield: 53.8 percent. The powder is detected by X-ray powder diffraction, has no characteristic peak and is amorphous.

Of the resulting product ¹ H-NMR is shown below, and nuclear magnetic data indicate that the molar ratio of the main component to L-lysine in the salt is 1: 1.

¹ H-NMR(400MHz，CD3OD)δ7.96(s,1H),7.80(s,1H),7.32-7.42(m,8H),6.87(d,2H),6.76(d,2H),6.39(d,1H),5.71(s,1H),4.42-4.51(m,1H),4.19(q,2H),3.55(t,1H),3.39(dd,1H),2.89-2.96(m,2H),2.77(dd,1H),1.81-1.90(m,2H),1.72-1.79(m,1H),1.64-1.72(m,2H),1.43-1.60(m,5H),1.01(d,3H),0.77-0.83(m,2H),0.48-0.56(m,2H).

Example 1 preparation of form II

The compound represented by the formula (II) (200mg, 0.4mmol, purity: 98.2%) and methanol (10mL) were added to a reaction flask, heated and stirred to dissolve, and then a suspension of L-lysine (58.1mg, 0.4mmol) and methanol (5mL) was added, and after stirring for 2 hours, the mixture was slowly cooled to room temperature and stirred, filtered, rinsed with methanol, and dried to obtain the product (167mg, pale yellow solid, purity: 99.6%), yield: and (4) 64.7%. The XRPD spectrum is shown in figure 1, the DSC spectrum is shown in figure 2, and the TGA spectrum is shown in figure 3 by X-powder diffraction detection; the characteristic peak positions are shown in the following table:

table 1, II crystal form characteristic peaks

Example 2 preparation of form II

The compound of formula (II) (70mg, 0.14mmol) and ethyl acetate (1mL) were added to a reaction flask, heated with stirring to dissolve, L-lysine (20mg, 0.14mmol) was added, after stirring for 1 hour, slowly cooled to room temperature with stirring, filtered, rinsed with a small amount of ethyl acetate, and dried to give the product (54mg, pale yellow solid), yield: 60 percent, and the product is determined to be II crystal form by X-ray powder diffraction detection.

Example 3 preparation of form II

Compound of formula (I) (20mg, 0.031mmol, from comparative example 2) and ethanol (2mL) were added to a reaction flask, heated with stirring to dissolve, seeded (example 1), stirred to crystallize and cooled to room temperature, filtered, rinsed with a small amount of ethanol, and dried to give compound (10.1mg, yellow solid), yield: 50.5 percent, and the product is determined to be II crystal form by X-ray powder diffraction detection.

Example 4 preparation of form II

The compound of formula (I) (100mg, 0.154mmol from comparative example 2) and methanol (2mL) were added to a reaction flask, heated to 60 ℃ and stirred, water (0.3mL) was added dropwise until the solution was clear, then slowly cooled to room temperature, seeded (example 1), stirred, filtered, rinsed with a small amount of methanol, and dried to give the compound (50mg, yellow solid) in yield: 50 percent, and the product is determined to be II crystal form by X-ray powder diffraction detection.

Example 5 preparation of form II

Adding a mixed solvent of a compound (100mg, 0.198mmol, purity: 98.47%), ethanol and water (V: 8:1, 2mL) shown in the formula (II) into a reaction bottle, heating to dissolve, adding L-lysine (31.93mg, 0.217mmol), stirring to react for 2-3 hours, crystallizing by adopting a heat-cold circulation mode (namely, stirring for 0.5-1 hour at 60 ℃, slowly cooling to 40 ℃, stirring for 0.5-1 hour, heating to 50 ℃, stirring for 0.5-1 hour, slowly cooling to 30 ℃, stirring for 0.5-1 hour, heating to 40 ℃, stirring for 0.5-1 hour, then slowly cooling to room temperature, stirring, filtering, drying to obtain a product (78mg, light yellow solid, purity: 98.61%), and yield: 61 percent, and the product is determined to be II crystal form by X-ray powder diffraction detection.

Example 6 preparation of form II

The compound of formula (I) (180mg, 0.28mmol, from comparative example 2) was dispersed in acetone (2mL) and water (0.04mL), slurried and stirred at room temperature for 7 days, filtered, and dried to give the product (107mg, yellow solid) in yield: 59.4 percent, and the product is determined to be II crystal form by X-ray powder diffraction detection.

Example 7 preparation of form II

A compound represented by the formula (II) (150mg, 0.30mmol, purity: 98.2%) and isopropanol (2.2mL) were added to a reaction flask, heated with stirring to dissolve, a solution of L-lysine (42.7mg, 0.29mmol) in methanol (1.2mL) was added, slowly cooled to room temperature with stirring to crystallize, filtered, and dried to give the product (119mg, yellow solid, purity: 98.54%), yield: 62 percent, and the product is determined to be II crystal form by X-ray powder diffraction detection.

Example 8 preparation of form II

Dissolving a compound (101mg, 0.2mmol, purity: 96.23%) shown in the formula (II) in acetonitrile (0.5mL), adding L-lysine (66.3mg, 0.453 mmol)/acetonitrile (3.5mL), heating and stirring for reaction for 20-40 minutes, then slowly cooling to room temperature and stirring for crystallization, filtering, and drying to obtain a product (70mg, light gray solid, purity: 99.08%), yield: 53.8 percent. Detecting by X-powder diffraction, wherein XRPD pattern is shown in figure 8, and is defined as I crystal form;

form I (50mg, 0.077mmol) was added to a mixed solvent of acetonitrile and water (V: V ═ 50:1, 1.02mL), slurried and stirred at room temperature for 7 days, filtered, and dried to give the title product (48mg, yellow solid) in yield: 96 percent. The crystal form II is determined by X-ray powder diffraction detection.

Example 9 solubility assay of form II of the present disclosure in water

Weighing a proper amount of sample, adding a proper volume of solvent, adding a stirrer, stirring to prepare a saturated solution, stirring for 18-24 h, measuring the pH value, filtering or centrifuging, and taking the supernatant to test the solubility.

Test results

TABLE 1 solubility comparison of crystalline form II of the present disclosure with compounds of formula (II)

And (4) test conclusion:

the solubility data in table 1 show that the solubility of the crystalline form of compound II of formula (I) is better than the product of comparative example 1.

Example 10 hygroscopicity study of the form II of the present disclosure

TAQ5000VSA is adopted, the humidity is 10-90% at 25 ℃, the step is 10%, the mass change is less than 0.01% within 10000min, and the process is circulated for two circles.

Results of the experiment

TABLE 2 investigation results of crystal form II hygroscopicity of the present disclosure

And (4) experimental conclusion:

as can be seen from table 2, the water absorption increases with the increase of humidity between 10.0% RH and 90.0% RH at 25 ℃, the weight change is 1.819%, less than 2% but not less than 0.2%, and the sample of the form II of the compound of formula (I) is slightly hygroscopic; the desorption process and the adsorption process of the sample are basically overlapped in the process of changing the humidity of 10-85 percent; the spectrum of DVS is shown in figure 4, and the contrast of X-ray powder diffraction before and after DVS shows that the crystal form before and after DVS is not transformed (see figure 5).

Example 11, crystal form II influencing factor experiment of the present disclosure

The product of comparative example 1 and the crystal form II of the compound of formula (I) were each left open and laid flat, and the stability of the samples under heating (40 ℃ C., 60 ℃ C.), illumination (45000Lux), high humidity (RH 75%, RH 90%) was examined for a sampling period of 5 days.

Results of the experiment

Table 3, product of comparative example 1 and Experimental results on Crystal form influencing factors of Compound II of formula (I)

The experimental conclusion is that:

the influence factor experiment results of table 3 show that: the crystal form II of the compound shown in the formula (I) has stability which is obviously superior to that of the product in the comparative example 1 after being placed for 5 days under the conditions of high temperature, high humidity, illumination and other influencing factors.

Example 12 long-term accelerated stability test of crystalline form II of the present disclosure

Form II of the compound of formula (I) (example 1) was subjected to a long term accelerated stability study for 3 months.

Results of the experiment

Table 4, results of long term accelerated stability experiments for the crystalline form of compound II of formula (I):

and (4) experimental conclusion:

the long term accelerated stability test results from table 4 show that: the XRPD peak pattern of the crystalline form of compound II of formula (I) placed for 3 months under long term accelerated stability conditions was essentially unchanged, the crystalline form was stable (long term XRPD see fig. 6, accelerated XRPD see fig. 7), and the physicochemical properties of the crystalline form of compound II of formula (I) placed under long term stability conditions (25 ℃, 60% RH) and accelerated stability conditions (40 ℃, 75% RH) were stable for 3 months, combining sample purity data.

Example 13 in vivo pharmacokinetic experiments of the compound shown in the crystal form II (example 1) and the formula (II) of the disclosure in dogs

The method is characterized in that a Beagle dog is taken as a test animal, the LC/MS/MS method is applied to determine the drug concentration in plasma of the Beagle dog at different moments after the Beagle dog is intragastrically administered with the compound shown in the formula (II) and the compound shown in the formula (I), the pharmacokinetic behaviors of the crystal form II and the free state in the Beagle dog body are researched, and the pharmacokinetic characteristics of the crystal form II and the free state in the Beagle dog body are evaluated.

And (3) testing the sample: the compound shown in the formula (II) and the crystal form II of the compound shown in the formula (I) can be prepared by a method shown in a comparative example 1 or an example 1.

Test animals: healthy adult Beagle dogs, 6 females, were divided into 2 groups and purchased from beijing mastic biotechnology limited.

Preparing the medicine: 0.5 percent of CMC-Na is ultrasonically processed into uniform suspension, the preparation concentration is 0.4mg/mL, and the uniform suspension is used for oral administration.

Administration: healthy adult Beagle dogs, 6 females, fasted overnight and gavaged at a 5mL/kg dose volume.

The method comprises the following steps:

6 healthy adult Beagle dogs and females are fasted overnight and then subjected to intragastric administration, blood is collected for 1mL before and after administration at 0.5, 1,2, 4, 6, 8, 12 and 24h through jugular venipuncture, heparin sodium is subjected to anticoagulation, blood plasma is separated by centrifugation at 3500rpm for 10min, and the blood plasma is stored at the temperature of-80 ℃. And (3) determining the content of the compound to be tested in the blood plasma of Beagle dogs after the intragastric administration of different compounds by an LC/MS/MS method.

Results of the experiment

TABLE 5 Beagle dog drug replacement evaluation results (po:2.0mg/kg)

Wherein, T _1/2 Is the half-life; AUC _last Area under curve 0 → t when the medicine is taken; cl is the clearance rate; vz _ F is the apparent distribution volume.

The experimental conclusion is that:

from the experimental results of table 5, it can be seen that, compared with the compound represented by formula (II), the half-life of the compound represented by formula (I) in the crystal form II is longer, the clearance rate is lower, and the exposure amount is higher than that of the compound represented by formula (II), which indicates that the pharmacokinetic properties of the compound represented by formula (I) in the crystal form II are good.

Claims

1. A crystalline form II of a compound of formula (I) characterized by: an X-ray powder diffraction pattern expressed by the angle of diffraction 2 theta is shown in figure 1,

2. the crystalline form II according to claim 1, characterized wherein the 2 Θ angle error range is ± 0.20.

3. A process for preparing the crystalline form II of any one of claims 1-2, wherein the process is selected from:

the method comprises the following steps: adding a compound shown in a formula (II) into a solvent (I), stirring for dissolving or heating for dissolving, adding L-lysine or a solution/suspension of the L-lysine and the solvent (I), and stirring for crystallizing, wherein the solvent (I) is at least one selected from an alcohol solvent, an ester solvent or a mixed solvent of the alcohol solvent, the ester solvent and water,

the second method comprises the following steps: adding a compound shown in a formula (I) into a solvent (II), stirring for dissolving or heating for dissolving, and then stirring for crystallization, wherein the crystallization mode is selected from room temperature crystallization, cooling crystallization, solvent volatilization crystallization or crystal seed addition for crystallization induction, the solvent (II) is selected from at least one of alcohol solvents or mixed solvents of alcohol solvents and water,

or, the third method: adding a compound shown in a formula (I) into a solvent (III), and pulping, wherein the solvent (III) is selected from a mixed solvent of a ketone solvent, a nitrile solvent and water.

4. The production method according to claim 3, wherein the solvent (I) is at least one selected from methanol, ethanol, isopropanol, ethyl acetate or a mixed solvent thereof with water.

5. The method according to claim 3, wherein the crystallization is induced by adding a seed crystal.

6. The production method according to claim 3, wherein the solvent (II) is at least one selected from methanol, ethanol or isopropanol or a mixed solvent thereof with water.

7. The preparation process according to claim 3, wherein the solvent (III) is selected from acetone/water, acetonitrile/water.

8. A pharmaceutical composition prepared from the crystalline form II of any one of claims 1-2 and one or more pharmaceutically acceptable carriers, diluents, or excipients.

9. Use of the crystalline form il according to any one of claims 1-2, or a pharmaceutical composition of the crystalline form il according to claim 8, for the manufacture of a medicament for the treatment of a disease or disorder mediated or dependent on an estrogen receptor.

10. The use according to claim 9, wherein the estrogen receptor mediated or dependent disease or condition is selected from the group consisting of cancer, central nervous system defects, cardiovascular system defects, immune and inflammatory diseases, susceptible infections, metabolic defects, psychiatric defects, or reproductive defects; the cancer is selected from breast cancer, endometrial cancer, cervical cancer, skin cancer, prostate cancer, ovarian cancer, fallopian tube tumors, hemophilia or leukemia.

11. The use according to claim 10, wherein the cancer is selected from breast, ovarian, endometrial, prostate or uterine cancer.

12. The use of claim 10, wherein the cancer is breast cancer.