TW201829398A - Protein tyrosine kinase modulators salt, crystallographic forms, and uses thereof - Google Patents

Abstract

The present invention relates to N- (5- (5-chloro-4- (2- (2- (dimethylamino) -2-oxoacetyl) phenylamino) pyrimidin-2-ylamino) -4-methoxy-2- (4-methylpiperazin-1-yl) phenyl) acrylamide succinate (the compound of Formula I) and new crystalline forms thereof, processes for their preparation, their uses in therapy and the pharmaceutical compositions containing them.

Description

 Tyramine protein kinase modulator, crystal form and use thereof  

The present invention relates to N- (5-(5-chloro-4-(2-(2-(dimethylamino))-2-oxoethyl)phenylamino)pyrimidin-2-ylamino)- 4-methoxy-2-(4-methylpiperazin-1-yl)phenyl)propenylamine succinate and novel crystal form thereof, compound, new crystal form preparation method and pharmaceutical composition thereof, Compounds, novel crystal forms, and pharmaceutical compositions that modulate kinases, particularly, modulate the activity of a selective mutant epidermal growth factor receptor (EGFR) kinase.

Epidermal growth factor receptor (EGFR, Erb-Bl) is a family of proteins involved in the proliferation of normal and malignant cells (Artega, C. L. J. Clin Oncol 19, 2001, 32-40). Excessive expression of the epidermal growth factor receptor (EGFR) in at least 70% of human cancers (Seymour, LK, Curr Drug Targets 2, 2001, 117-133), such as non-small cell lung cancer (NSCLC), breast cancer, glial Tumor, head and neck squamous cell carcinoma and prostate cancer, etc. (Raymond et al., Drugs 60 Suppl 1, 2000, discussion 41-2; Salomon et al., Crit Rev Oncol Hematol 19, 1995, 183-232; Voldborg et al , Ann Oncol 8, 1997, 1197-1206). The epidermal growth factor receptor tyrosine kinase (EGFR-TK) is widely recognized as a very attractive target because the designed compounds are specifically capable of binding to tyrosine kinase and inhibiting its activity, blocking its A signal transduction pathway in cancer cells that acts as a diagnostic or therapeutic agent. For example, Erlotinib and Gefitinib.

Although gefitinib/erlotinib has initial clinical benefit in the treatment of EGFR-mutant NSCLC patients, erlotinib and gefitinib have limited efficacy in the treatment of various lung cancer patients. When erlotinib or gefitinib is used to treat a variety of lung cancer patients (without the presence/absence of activated (mutant) EGFR), the likelihood of tumor shrinkage (response rate) is only 8%-10%. The median tumor progression time was approximately 2 months (Shepherd et al NEJM 2004, Thatcher et al. Lancet 2005).

In addition, most patients eventually develop advanced cancer when treated with gefitinib/erlotinib. Preliminary studies have shown that secondary EGFR mutations in patients with cancer recurrence, T790M, cause gefitinib and erlotinib to be ineffective in inhibiting EGFR kinase activity (Kobayashi et al NEJM 2005 and Pao et al PLOS Medicien 2005).

Rociletinib (CO-1686) is a novel orally administered kinase inhibitor that specifically targets EGFR mutants including T790M and exhibits little activity against wild-type receptors. In EGFR-mutated NSCLC tumor xenografts and transgenic models, oral administration of CO-1686 alone can cause tumor regression. However, high doses of CO-1686 alone may cause cardiac side effects in patients.

Therefore, there is still a need to develop more effective EGFR target drugs that inhibit EGFR T790M. This drug is clinically more effective and is more responsive to cancer patients as a therapeutic drug.

The present inventors have disclosed a group of heterocyclic pyrimidine compounds which modulate mutant selective epidermal growth factor receptor (EGFR) and ALK kinase activity in the patent application WO2015/003658 (protein tyrosine kinase modulator and its application method), In view of regulating cell activity, such as proliferation, differentiation, programmed cell death, migration, and chemical attack, they are among the best protein tyrosine kinase modulators. More specifically, such heterocyclic pyrimidine compounds are capable of selectively inhibiting, modulating and/or modulating kinase receptors, particularly various EGFR mutant activities associated with changes in cellular activity mentioned above.

Among the heterocyclic pyrimidine compounds mentioned, N- (5-(5-chloro-4-(2-(2-(dimethylamino))-2-oxoethyl)phenylamino)pyrimidine- The structure of 2-aminoamino)-4-methoxy-2-(4-methylpiperazin-1-yl)phenyl)propenylamine is disclosed in Example 74 of WO 2015/003658.

It is well known that the physicochemical properties of solid pharmaceutical active ingredients (APIs) are critical to the development of pharmaceutical products, as they may have an impact on the bioavailability, stability and processing properties of the active ingredients and the corresponding pharmaceutical forms.

It is known that in many cases the active ingredient may be present in crystalline or amorphous form in the solid state, and for crystalline forms, it may exist in various solvates and polymorphs.

Thus, a polymorphic material consists of a material that crystallizes into one or more crystalline forms, each crystal form being characterized by a different arrangement of molecules in the crystal lattice, and the ability to produce a solvate comprising a stoichiometric basis, at a precise location Water or solvent molecules are incorporated into the crystal lattice.

Different polymorphs and solvates may have different solubilities, different stability, different hygroscopicity and different mechanical properties such as filterability and flowability.

Although solubility is important for the bioavailability of a drug, other physicochemical and mechanical properties are important for determining the stability and processing properties of the active ingredient and the drug form, and thus may have a significant impact on the quality and cost of the drug. Depending on the therapeutic use, the route of administration, and the type of formulation, it may be desirable to impart different physicochemical properties to the same active ingredient to make it suitable for a variety of formulation requirements.

Polymorphs may therefore be an advantageous opportunity to meet these requirements. For example, in the case of an oral rapid-dose pharmaceutical or parenteral formulation, the importance of the solubility of the active ingredient will be manifested in determining the efficacy or even the likelihood of using the route of administration.

The absorption of oral drugs is determined by two fundamental factors, permeability, the ability to spread across the gastrointestinal wall, and solubility, the ability to dissolve in gastrointestinal fluids.

Taking these two factors into consideration, N -(5-(5-chloro-4-(2-(2-(dimethylamino))-2-oxoethyl)phenylamino)pyrimidin-2-yl Amino)-4-methoxy-2-(4-methylpiperazin-1-yl)phenyl)propenylamine has high permeability and low solubility, so solubility determines the basis of its absorption. The preparation of pharmaceutically acceptable salts generally represents a means of increasing the solubility of the sparingly soluble product and, in the case of good permeability of the product, is a suitable means of increasing its bioavailability.

However, it is not always feasible to obtain a salt having suitable properties such as solubility, stability and processability. Salts of organic compounds can produce polymorphs and solvates, and occasionally may be crystalline forms of suitable salts or solvates having the foregoing properties, which will allow for the preparation of pharmaceutical formulations that are suitable for the preparation. For example, in the preparation of the active ingredient and the corresponding pharmaceutical formulation, the stability of the crystalline form under various storage conditions is the basic condition for ensuring the quality, uniformity and consistency of the drug.

In most cases, in the preparation process, the hygroscopicity or low stability of the crystalline form is utilized to avoid the use of specific known precautions to greatly reduce manufacturing costs. The stability of the crystal form to mechanical stress is important in the conventional processes typically used in the pharmaceutical preparation process, such as milling, which is necessary to achieve a suitable particle size to meet formulation (fluidity) and dissolution requirements; It is necessary to ensure that the uniformity of the active ingredients in the formulation product is necessary; as well as compression, which is necessary for the preparation of the tablet system.

The determination and characterization of the crystal form can often be an unusual process (Giron Danielle, Monitoring polymorphism of drugs, an on-going challenge-part 2. American Pharmaceutical Review (2008), 11(3), 86-90). The use of many complementary analytical techniques, such as X-ray diffraction, calorimetry, and vibrational spectroscopy, allows for the definitive determination and characterization of a given crystal form in most cases.

Thermogravimetric analysis (TGA), usually combined with differential scanning calorimetry (DSC), is very useful for demonstrating the presence of hydrates or solvates. DSC is also an important technology to characterize polymorphs and associated thermal properties.

The hygroscopicity of compounds is studied in the case of hydrates, and dynamic vapor adsorption (DVS) is an important technique. Whether it is a polymorph or a solvate, the method of characterizing crystallization can be selected from X-ray spectroscopy. X-ray powder diffraction (XRPD) This relatively simple technique can clearly determine crystal form and relative crystallinity in most cases (Harry G. Brittain, X-ray powder diffraction of pharmaceutical materials, American Pharmaceutical Review 2002, 5(1), 74-76).

After proper calibration, XRPD can be used to determine the purity of the polymorph and has very high sensitivity (Stephen R. Byrn, Regulatory aspects of X-ray powder diffraction, American Pharmaceutical Review 2005, 8(3), 55-59) In the process of identifying and characterizing polymorphs, the technique is capable of detecting the presence of other crystal forms with a sensitivity of typically about 5-10%.

The best method for identifying and characterizing the crystal form is the single crystal X-ray diffraction (SC-XRD) pattern. It can identify the type and size of the unit cell, characterize the type of crystal form, and thus is the most suitable way to determine the polymorph or solvent compound, and in the case of salt, to clearly clarify its stoichiometry and understand its Attributes.

Despite considerable technological advances, the main limitation of this technique remains the possibility of obtaining a crystal form to be analyzed of suitable size and with a limited number of defects, which is not always easy or even impossible.

The object of the present invention is to provide a novel crystalline form of the compound of formula I wherein the compound is N- (5-(5-chloro-4-(2-(2-(dimethylamino))-2-oxoethyl) Phenylamino)pyrimidin-2-ylamino)-4-methoxy-2-(4-methylpiperazin-1-yl)phenyl)propenylamine succinate.

The present invention provides a crystalline form of the compound of Formula I, referred to as Form A, which has a characteristic peak of diffraction angle 2θ of about 7.5°, 13.2°, and 21.0°±0.2°.

The invention further provides a preferred embodiment of Form A.

Ideally, the X-ray powder diffraction pattern has characteristic peaks expressed by the interplanar spacing at 11.8 Å, 6.7 Å, and 4.2 Å.

Desirably, the X-ray powder diffraction pattern has characteristic peaks of diffraction angles 2θ of about 4.8°, 7.5°, 13.2°, 19.0°, 19.6°, and 21.0°±0.2°.

Ideally, the X-ray powder diffraction pattern has characteristic peaks expressed by the interplanar spacing at 18.6 Å, 11.8 Å, 6.7 Å, 4.7 Å, 4.5 Å, and 4.2 Å.

Desirably, the X-ray powder diffraction pattern has characteristic peaks of diffraction angles 2θ of about 4.8°, 7.5°, 13.2°, 19.0°, 19.6°, 21.0°, 22.5°, and 26.1°±0.2°.

Ideally, the X-ray powder diffraction pattern has characteristic peaks expressed by the interplanar spacing at 18.6 Å, 11.8 Å, 6.7 Å, 4.7 Å, 4.5 Å, 4.2 Å, 3.9 Å, and 3.4 Å.

The X-ray powder diffraction pattern depicted in Figure 1 is summarized in Table 1.

Desirably, Form A has a melting point of about 140.0 ° C to 145.0 ° C.

More desirably, the melting point of Form A is about 141.0 ° C to 143.0 ° C.

Still more desirably, the melting point of Form A is about 142.0 °C.

Ideally, the purity of Form A 75%.

Ideally, the purity of Form A 80%.

Ideally, the purity of Form A 85%.

Ideally, the purity of Form A 90%.

Ideally, the purity of Form A 95%.

Ideally, the purity of Form A 99%.

Ideally, the purity of Form A 99.5%.

Ideally, the X-ray powder diffraction pattern of Form A is shown in Figure 1.

The present invention also provides a crystalline form of the compound of Formula I, referred to as Form B, having a characteristic peak having a diffraction angle 2θ of about 6.4°, 17.9°, and 19.1°±0.2°.

Ideally, the X-ray powder diffraction pattern has characteristic peaks expressed by the interplanar spacing at 13.6 Å, 5.0 Å, and 4.7 Å.

Desirably, the X-ray powder diffraction pattern has characteristic peaks of diffraction angles 2θ of about 6.4°, 17.9°, 19.1°, 20.0°, 21.5°, and 23.0°±0.2°.

Ideally, the X-ray powder diffraction pattern has characteristic peaks expressed by the interplanar spacing at 13.6 Å, 5.0 Å, 4.7 Å, 4.5 Å, 4.1 Å, and 3.9 Å.

Desirably, the X-ray powder diffraction spectrum has characteristic peaks of diffraction angles 2θ of about 6.4°, 11.8°, 14.5°, 15.9°, 17.9°, 19.1°, 20.0°, 21.5°, 23.0°, and 24.1°±0.2°. .

Ideally, the X-ray powder diffraction pattern has characteristic peaks expressed by the interplanar spacing at 13.6 Å, 7.5 Å, 6.1 Å, 5.6 Å, 5.0 Å, 4.7 Å, 4.5 Å, 4.1 Å, 3.9 Å, and 3.7 Å.

The X-ray powder diffraction pattern depicted in Figure 2 is summarized in Table 2.

Desirably, Form B has a melting point of about 133.0 ° C to 137.0 ° C.

More desirably, the melting point of Form B is about 134.0 ° C to 136.0 ° C.

Still more desirably, the melting point of Form B is about 135.1 °C.

Ideally, the purity of Form B 75%.

Ideally, the purity of Form B 80%.

Ideally, the purity of Form B 85%.

Ideally, the purity of Form B 90%.

Ideally, the purity of Form B 95%.

Ideally, the purity of Form B 99%.

Ideally, the purity of Form B 99.5%.

Ideally, the X-ray powder diffraction pattern of Form B is shown in Figure 2.

The above crystalline form A or B is a relatively stable crystalline form composed of uniformly distributed fine particles having an average particle size (D ₉₀ ) of about 1 to 10 μm, and can be easily produced into a pharmaceutical product for clinical use.

In another aspect, the invention provides a process for the preparation of a crystalline form of a compound of formula I, which comprises the steps of: N- (5-(5-chloro-4-(2-(2-(dimethylamino))-2-oxo) Ethyl)phenylamino)pyrimidin-2-ylamino)-4-methoxy-2-(4-methylpiperazin-1-yl)phenyl)propenylamine (hereinafter referred to as free base) And succinic acid is mixed in a solvent and stirred for 8 to 96 hours to form a reaction mixture, N- (5-(5-chloro-4-(2-(2-(dimethylamino))-2-oxoethyl hydrazine) Phenylamino)pyrimidin-2-ylamino)-4-methoxy-2-(4-methylpiperazin-1-yl)phenyl)propenylamine succinate precipitated from the reaction mixture Come out to get the crystal form.

In some embodiments, the solvent is ethyl acetate or acetone.

In some embodiments, the free base and succinic acid are mixed in a molar ratio of 1:1.

In some embodiments, the free base and succinic acid are mixed in acetone at room temperature (RT) or at 50 ° C and the crystalline form is Form A.

In other embodiments, the free base and succinic acid are combined in acetone at about 50 ° C in crystalline form as Form A. In some embodiments, the free base and succinic acid are combined in ethyl acetate at about 50 ° C in crystalline form as Form B. The growth of the single crystal provided by the present invention under specific conditions depends on the particular thermodynamic and equilibrium properties of the crystallization process. Therefore, those of ordinary skill in the art will be aware of the kinetic and thermodynamic properties of the crystallization process of the crystal system formed. Under certain conditions (eg, solvent, temperature, pressure, and concentration of the compound of the invention), a particular crystalline form may be more stable than other crystalline forms or indeed a particular crystalline form may have a more stable beneficial power than other crystalline forms. learn. Other factors than kinetics such as time, impurity distribution, agitation, and presence or absence of seed crystals may also affect the crystal form.

The invention further provides for the use of Forms A and B.

The pharmaceutical compositions comprise a therapeutically effective amount of one or both of Form A or Form B of a compound of Formula I and a pharmaceutically acceptable inactive component.

The invention further provides a preferred embodiment of a pharmaceutical composition.

Desirably, the pharmaceutical composition comprises a therapeutically effective amount of Form A and/or B of the present invention and at least one other active ingredient.

Desirably, the pharmaceutical composition is for oral administration.

Desirably, the pharmaceutical composition is used as a tablet or capsule.

Desirably, the pharmaceutical composition comprises from 0.1% by weight to 99.9% by weight of Form A and/or B of the present invention.

Desirably, the pharmaceutical composition comprises from 1% by weight to 90% by weight of Form A and/or B of the present invention.

Desirably, the pharmaceutical composition comprises from 2% by weight to 80% by weight of Form A and/or B of the present invention.

Desirably, the pharmaceutical composition comprises from 5 wt% to 70 wt% of Form A and/or B of the present invention.

Desirably, the pharmaceutical composition comprises from 7.5 wt% to 60 wt% of Form A and/or B of the present invention.

Desirably, the pharmaceutical composition comprises from 10% to 50% by weight of Form A and/or B of the present invention.

Desirably, the pharmaceutical composition comprises from 10% by weight to 30% by weight of Form A and/or B of the present invention.

Such crystalline forms of the invention or the above pharmaceutical compositions are useful in the manufacture of a medicament for treating a disease in a subject.

The invention also provides some desirable technical means for the above uses.

In some embodiments, the medicament so prepared can be used to treat or prevent cancer, cancer metastasis, cardiovascular disease, immunological or ocular condition, or to delay or prevent the onset or progression of such disease.

In some embodiments, the medicament so prepared is useful for inhibiting kinases.

In some embodiments, the kinase comprises EGFR, ALK, ALK fusion protein, Flt3, Jak3, Blk, Bmx, Btk, HER2 (ErbB2), HER4 (ErbB4), Itk, Tec or Txk.

In some embodiments, the EGFR is a mutant EGFR; the aforementioned cancer is an EGFR-driven cancer, and the aforementioned EGFR-driven cancer is characterized by the presence of one or more mutations selected from the group consisting of: (i) L858R, (ii) T790M, (iii) L858R and T790M, (iv) delE746_A750 or (v) delE746_A750 and T790M.

In some embodiments, the EGFR-driven cancer is non-small cell lung cancer (NSCLC), glioblastoma, pancreatic cancer, head and neck cancer (eg, squamous cell carcinoma), breast cancer, colorectal cancer, epithelial cancer, ovarian cancer, Prostate cancer or adenocarcinoma.

In some embodiments, the ALK fusion protein is MEL4-ALK or NPM-ALK kinase.

In some embodiments, the subject is human.

The invention further provides a method of inhibiting kinase in a subject comprising administering Form A or B of the compound of Formula I or a pharmaceutical composition as described above.

The invention further provides some desirable technical means in relation to the above methods.

In some embodiments, the kinase comprises EGFR, ALK, ALK fusion protein, Flt3, Jak3, Blk, Bmx, Btk, HER2 (ErbB2), HER4 (ErbB4), Itk, Tec or Txk.

In some embodiments, the EGFR is a mutant EGFR, and the ALK fusion protein is a MEL4-ALK or NPM-ALK kinase.

The invention further provides a method of treating a disease in a subject comprising administering to the subject a crystalline form A or B of the compound of formula I or a pharmaceutical composition as described above.

The invention further provides some desirable technical means in relation to the above methods.

In some embodiments, the disease is caused by a kinase regulatory disorder, and the kinase comprises EGFR, ALK, ALK fusion protein, Flt3, Jak3, Blk, Bmx, Btk, HER2 (ErbB2), HER4 (ErbB4), Itk, Tec or Txk.

In some embodiments, the disease described is an EGFR-driven cancer, and the aforementioned EGFR-driven cancer is characterized by the presence of one or more mutations selected from the group consisting of: (i) L858R, (ii) T790M, (iii) L858R And T790M, (iv) delE746_A750 or (v) delE746_A750 and T790M.

In some embodiments, the EGFR-driven cancer is non-small cell lung cancer (NSCLC), glioblastoma, pancreatic cancer, head and neck cancer (eg, squamous cell carcinoma), breast cancer, colorectal cancer, epithelial cancer, ovarian cancer, Prostate cancer or adenocarcinoma.

In some embodiments, the ALK fusion protein is MEL4-ALK or NPM-ALK kinase.

In some embodiments, the subject is human.

"EGFR-driven cancer" refers to a cancer characterized by mutations in the EGFR gene that alter the biological activity of an EGFR nucleic acid molecule or polypeptide, including the specific mutations mentioned herein. EGFR-driven cancer can occur in any tissue, including brain, blood, connective tissue, liver, mouth, muscle, spleen, stomach, testicles, or trachea. EGFR-driven cancer includes non-small cell lung cancer (NSCLC), including squamous cell carcinoma, adenocarcinoma, bronchioloalveolar carcinoma (BAC), BAC with local infiltration, adenocarcinoma with BAC characteristics, and large cell carcinoma Multiple, neurological tumors, such as glioblastoma, pancreatic cancer, head and neck cancer (eg squamous cell carcinoma), breast cancer, colorectal cancer, epithelial cancer, including squamous cell carcinoma, ovarian cancer, prostate cancer, including EGFR-mediated Cancer.

An "EGFR mutant" or "mutant" comprises one or more deletions, substitutions or additions in the amino acid or nucleotide sequence of the EGFR protein or EGFR coding sequence. An EGFR mutant may also comprise one or more deletions, substitutions or additions, or fragments thereof, as long as the mutant retains or increases tyrosine kinase activity compared to wild-type EGFR. In a particular EGFR mutation, the kinase or phosphorylation activity can be increased relative to wild-type EGFR (eg, at least 5%, 10%, 15%, 20%, 30%, 40%, 50%, 60%, 70%, 80). %, 90% or even 100%). Specific EGFR mutants are as described in the specification wherein mutations are provided relative to the position of the amino acid in human EGFR as described in the sequence provided in NCBI GenBank Reference Sequence: NP_005219.2.

As used herein, the term "inhibiting cell proliferation expressing an EGFR mutant" means moderately slowing, stopping or reversing the growth rate of cells expressing EGFR in vitro or in vivo. Ideally, when measured using a suitable method for measuring cell growth rate (e.g., the cell growth assay described herein), the growth rate is at least 10%, 20%, 30%, 50%, or even 70% slower. The EGFR mutant can be any of the EGFR mutants described in this specification.

Furthermore, the present invention provides crystalline form A or B or a pharmaceutical composition of the above formula, for use in the treatment of cancer, prevention of cancer metastasis or treatment of cardiovascular, immunological or ocular diseases.

The invention further provides a method of treating a patient having a condition mediated by protein kinase activity, the method comprising administering to the patient a therapeutically effective amount of a crystalline form as described herein. Examples of protein kinases include the mutants EGFR, KDR, Tie-2, Flt3, FGFR3, AbI, Aurora A, c-Src, IGF-IR, ALK, c-MET, RON, PAK1, PAK2, and TAK1.

In some embodiments, the condition mediated by protein kinase activity is cancer.

Further, a method of treating cancer in a mammal comprising administering to a mammal in need of such treatment an effective amount of a crystalline form of a compound of formula I or a pharmaceutical composition as described above.

Examples of cancer include, but are not limited to, lung cancer, breast cancer, colorectal cancer, kidney cancer, pancreatic cancer, head and neck cancer, hereditary papillary renal cell carcinoma, childhood hepatocellular carcinoma, gastric cancer, sarcoma, fibrosarcoma, osteoma, melanin Tumor, retinoblastoma, rhabdomyosarcoma, glioblastoma, neuroblastoma, teratocarcinoma, hematopoietic malignancies and malignant ascites.

The amount of the active ingredient or compound to be administered is determined according to the individual needs of the subject, the route of administration, the severity of the disease, the administration schedule, and the evaluation and judgment of the designated physician. Preferably, the effective dosage is, based on the active compound, about 0.01 to 120 mg per kilogram of body weight; or more desirably, a single dose or a single dose is 1 to 50 mg per kilogram of body weight per day. In some cases, it is more appropriate to apply the lower limit of the above dosage range, while in other cases, higher doses may be used without causing harmful side effects.

Another aspect of the invention provides Form A or Form B of the compound of Formula I, which may be in the range of 25-2100 mg/day, administered at a frequency of 1-3 times per day; ideally a dose of 75- 1200 mg / day, the frequency of administration is 1-3 times a day; more desirably, the dose is 75-1200 mg / day, the frequency of administration is 2-3; further more desirably, the dose is 100-200 mg / day, the frequency of administration is daily 2-3 times.

The crystal form of the invention is approximately pure.

The term "approximately pure" as used herein means that the compound of formula I present in the crystalline form of the invention is at least 85% by weight, desirably at least 95% by weight, more preferably at least It is 99% by weight.

In the present invention, "X-ray powder diffraction pattern as shown in Fig. 1" means an X-ray powder diffraction pattern of a main peak as shown in Fig. 1, wherein the main peak means that the relative intensity is larger than the highest peak in Fig. 1 ( Its relative intensity is specified as 10% of 100%), ideally greater than 30%. Similarly, in the present invention, "X-ray powder diffraction pattern as shown in FIG. 2" means an X-ray diffraction pattern of a main peak as shown in FIG. 2, wherein the main peak means that the relative intensity is larger than that in FIG. 10% of the peak (the relative intensity is specified as 100%), ideally greater than 30%.

As used herein, the term "therapeutically effective amount" refers to an amount of a compound that is sufficient to affect such treatment when administered to a subject to treat at least one clinical condition of the disease or condition. The "therapeutically effective amount" may vary depending on the compound, the disease and its severity, the age, weight, etc. of the subject to be treated. Suitable amounts for any given situation will be apparent to those of ordinary skill in the art, or may be determined by routine experimentation. In the context of combination therapy, "therapeutically effective amount" means the total amount effective to treat a disease, disorder or combination of subjects.

A pharmaceutical composition comprising a compound of the invention can be administered to a patient in need of treatment by oral, inhalation, rectal, parenteral or topical administration. For oral administration, the pharmaceutical composition may be a conventional solid preparation such as a tablet, a powder, a granule, a capsule or the like; a liquid preparation such as an aqueous or oily suspension or other liquid preparation such as a syrup, a solution, a suspension, etc.; For external administration, the pharmaceutical composition may be a solution, an aqueous solution, an oil suspension, a lyophilized powder or the like. Desirably, the preparation of the pharmaceutical composition is selected from the group consisting of a tablet, a coated tablet, a capsule, a suppository, a nasal spray or an injection, more preferably a tablet or a capsule. The pharmaceutical composition can be administered in a single unit of precise dosage. Furthermore, the pharmaceutical composition may further comprise other active ingredients.

All formulations of the pharmaceutical compositions of the invention may be prepared by conventional methods in the pharmaceutical arts. For example, the active ingredient may be combined with one or more excipients, followed by the preparation.

"Pharmaceutically acceptable inactive ingredient" means a conventional pharmaceutical carrier suitable for the desired pharmaceutical preparation, such as a diluent; a carrier such as water, various organic solvents, etc.; a filler such as starch, sucrose, etc.; a binder such as a cellulose derivative , alginate, gelatin and polyvinylpyrrolidone (PVP); wetting agents such as glycerin; disintegrating agents such as agar, calcium carbonate and sodium hydrogencarbonate; absorption enhancers such as quaternary ammonium compounds; surfactants such as sixteen Alkanol; absorption carrier such as kaolin and soap clay; lubricants such as talc, calcium stearate, magnesium stearate, polyethylene glycol, and the like. Further, the pharmaceutical composition also contains other pharmaceutically acceptable excipients such as dispersing agents, stabilizers, thickeners, complexing agents, buffers, penetration enhancers, polymers, fragrances, sweeteners and dyes. Desirably, the excipient is suitable for the desired formulation and type of administration.

The term "disease" or "condition" or "symptom" means any disease, discomfort, symptom or indication.

The crystal form provided by the present invention is a relatively stable crystal form composed of uniformly distributed fine particles having an average particle size (D ₉₀ ) of about 1 to 10 μm, and can be easily produced into a pharmaceutical product for clinical use. Furthermore, the medicaments prepared by the technical means of the present invention can be used for the treatment or prevention of cancer, cancer metastasis, cardiovascular diseases, immunological disorders or ocular disorders, or for delaying or preventing the onset or progression of such diseases.

Fig. 1 shows an XRPD pattern of Form A.

Fig. 2 shows an XRPD pattern of Form B.

Fig. 3 is a graph showing a dynamic vapor sorption (DVS) pattern of Form A.

Fig. 4 shows an XRPD overlap pattern of Form A before and after DVS.

Fig. 5 shows an XRPD overlap pattern of Form A before and after storage.

The invention is further illustrated by, but not limited to, the following examples. In the examples of the present invention, the described techniques or methods are conventional techniques or methods in the art unless otherwise specifically stated.

Temperature is measured in degrees Celsius (°C). The reaction or experiment is carried out at room temperature unless otherwise stated. The room temperature is approximately 25 ± 10 °C.

Abbreviations: ACN: acetonitrile; DMF: dimethylformamide; DSC: differential scanning calorimetry; DVS: dynamic vapor adsorption; EGFR: epidermal growth factor receptor; EtOAc: ethyl acetate; ¹ H-NMR: ¹ H NMR; HPLC: high performance liquid chromatography; RT: room temperature; RH: relative humidity; TGA: thermogravimetric analysis; XRPD: X-ray powder diffraction.

Example 1 Preparation of Form A

The free base was prepared by the method described in Example 74 of WO 2015/003658.

350 mg of free base and 70.1 mg of succinic acid were added to a 20 ml glass vial, followed by 10 ml of EtOAc in a glass vial. The mixture was stirred at RT for about 24 hours, then the wet cake was filtered and dried under reduced pressure at 50 ° C to give 314 mg of crystals. MS: m/z 710.3, mp: ca. 142.0.

¹ H NMR (400 MHz, DMSO) δ = 11.36 (s, 1H), 9.03 (s, 1H), 8.83 (s, 1H), 8.63 (s, 1H), 8.26 (s, 1H), 8.08 (s, 1H) ), 7.63 (d, J = 6.7, 1H), 7.52 (d, J = 8.2, 1H), 7.15 (t, J = 7.5, 1H), 6.88 (s, 1H), 6.61 (dd, J = 17.0, 10.2,1H), 6.17(d, J =17.0,1H), 5.71 (d, J = 10.9,1H), 3.78 (s, 4H), 3.01 (s, 4H), 2.90 (s, 8H), 2.61 ( s, 4H), 2.41 (s, 5H), 2.30 (s, 3H), 0.85 (d, J = 6.6, 0H).

Example 2 Preparation of Form A

The free base was prepared by the method described in Example 74 of WO 2015/003658.

350 mg of free base and 70.1 mg of succinic acid were added to a 100 ml glass vial, followed by 20 ml of EtOAc in a glass vial. The mixture was stirred at RT for about 96 hours, then the wet cake was filtered and dried under reduced pressure at 50 ° C to afford 317 g of crystals. MS: m/z 710.3, mp: about 141.0 ° C - 143.0 ° C.

Example 3 Preparation of Form A

The free base was prepared by the method described in Example 74 of WO 2015/003658.

About 340.1 g of the free base and 71.1 g of succinic acid were added to the reactor, followed by the addition of 27.2 L of acetone to the reactor. The mixture was stirred at 50 ° C for about 8 hours, cooled to room temperature, and then the wet cake was filtered and dried under reduced pressure at 50 ° C to give 285.6 g of crystal form A. MS: m/z 710.3, mp: about 141.0 ° C - 143.0 ° C.

Example 4 Preparation of Form B

The free base was prepared by the method described in Example 74 of WO 2015/003658.

0.1 mol of the free base and 0.1 mol of succinic acid were stirred in a suspension of 500 ml of EtOAc at 50 ° C for 33 hours. After filtration, the solid was dissolved in EtOAc, washed with water and dried The molar ratio of free base to succinic acid as determined by ¹ H NMR in Form B was 1:1.5. MS: m/z 769.3, mp: app. 135.1.

^{1 H NMR (400MHz, DMSO)} δ = 11.35 (s, 1H), 9.02 (s, 1H), 8.83 (s, 1H), 8.62 (s, 1H), 8.25 (s, 1H), 8.07 (s, 1H ), 7.62 (d, J = 6.9, 1H), 7.53 (s, 1H), 7.15 (t, J = 7.6, 1H), 6.88 (s, 1H), 6.61 (dd, J = 17.0, 10.2, 1H) , 6.16 (d, J = =18.6, 1H), 5.70 (d, J = 11.6, 1H), 4.03 (q, J = 7.2, 1H), 3.78 (s, 4H), 3.01 (s, 4H), 2.90 ( s, 8H), 2.64 (d, J = 24.6, 4H), 2.41 (s, 6H), 2.31 (s, 3H), 1.99 (s, 1H), 1.17 (t, J = 7.1, 1H).

Example 5 XRPD Analysis Results

The compound of formula I is characterized by XRPD. For XRPD analysis, a PANalytical Empyrean X-ray powder diffractometer was used. Table 3 lists the parameters used.

All other diffraction patterns described in the present invention are obtained in the same manner.

For the above crystal forms, only the main peaks (such as the most characteristic, significant, unique and/or reproducible peaks) are summarized; other peaks can be obtained from the diffraction spectrum by conventional methods. The main peaks described in the above crystal forms are reproducible and within the error range (specified value ± 0.2°).

It should be understood that the crystal form A or B provided by the present invention is not limited to a crystal form which provides the same X-ray powder diffraction pattern as the X-ray powder diffraction pattern shown in FIG. 1 or FIG. 2, and is provided as shown in FIG. 1 or FIG. Any crystal of substantially the same X-ray powder diffraction pattern falls within the scope of the invention.

Example 6 Hygroscopicity test

Form A is also characterized by the dynamic vapor sorption (DVS) given in Figure 3. As the DVS results show, when the relative humidity is changed from 0% to 80%, the water absorption of the sample of Form A is 0.782%, indicating that Form A has a slight hygroscopicity. There was no change in XRPD before and after the DVS test, indicating no change in crystal form after DVS test (Figure 4).

DVS was measured by an SMS (Surface Measurement System) DVS Intrinsic Dynamic Water Vapor Adsorber. The relative humidity was corrected by the tidal solution points of LiCl, Mg(NO ₃ ) ₂ and KCl at 25 °C. Table 4 lists the typical parameters of the DVS test.

All other DVS tests described in the present invention were obtained in the same manner.

Example 7 Solid State Stability Test

A sample of Form A was placed in an open pan for one week at 25 ° C / 60% RH and 40 ° C / 75% RH. XRPD and HPLC data of stable samples were collected and compared to the initial samples.

The results are summarized in Table 5 and the XRPD pattern is shown in Figure 5. XRPD or HPLC showed no change in solid form or purity in Form A at 25 ° C / 60% RH and 40 ° C / 75% RH, indicating that Form A has good physical and chemical stability.

The above results indicate that Form A has good stability, low hygroscopicity and better solubility.

Example 8 Pharmacokinetic Study of Form A and Free Base

Drugs and reagents: Crystalline A and free base are ground into fine particles. The sample content (purity) is not less than 99.0%. The hydroxymethylcellulose nano is medical grade.

Animal experiments: Wistar rats, male and female, 150-220 g each.

Pharmaceutical preparation: Weigh an appropriate amount of each substance and add 0.5% sodium carboxymethylcellulose. A suspension with a final concentration of 3.5 mg/ml was prepared.

Drug and sample collection: Each suspension was orally administered to fasted Wistar rats at a dose equivalent to 35 mg/kg of free base in a dose volume of 10 ml/kg. About 0.5-1.0 ml of plasma was collected in heparinized tubes at 1, 2, 3, 6, 10, and 24 hour intervals after administration, and stored at -20 ° C after centrifugation.

After purification, the samples were analyzed by HPLC. The chromatographic conditions were as follows: C18 decane bonded silica as the stationary phase, acetonitrile and water (0.1% formic acid) as the mobile phase, and the detection wavelength was 254 nm. The table below shows the area of each compound under the concentration-time curve. Table 6 summarizes the PK data for the crystalline form A and free base of the compound of formula I.

Example 9 Tumor Suppressive Effect on Human Tumor Xenografts in Nude Mice

This is a preliminary study comparing the crystalline form A of the present invention with free base to inhibit human H1975 (non-small cell lung cancer) xenograft tumors.

METHODS: Human lung adenocarcinoma cell line H1975 was purchased from the Basic Medical Research Cell Culture Center of the Chinese Academy of Medical Sciences. The cells were cultured in a humidified atmosphere containing 5% CO ₂ at 37 ° C in RPMI-1640 medium (containing 10% fetal bovine serum, 100 U/ml penicillin and 100 U/ml streptomycin).

H1975 non-small cell lung cancer cells were subcutaneously inoculated into the right axilla of NALB/C nude mice to produce tumor nodules. These nodules were collected, cut into 6 mm ³ pieces for routine inoculation, and implanted into each mouse for study. When the tumor grew to approximately 20 mm ³ (6-7 days), the animals were randomly divided into 3 groups of 6-9 each and their body weights were monitored. Each group consisted of a control group (no drug treatment), a crystal form A treatment group (30 mg/kg), and a free base treatment group (30 mg/kg). Oral administration once a day. The frequency of administration depends on the individual condition of tumor growth. Tumor volume [tumor volume (V) = tumor size (L) x tumor diameter (S) ² /2] was measured every 3 days using a caliper. Animals were sacrificed 24 hours after the last dose and weighed. Tumors are recovered and weighed to accurately calculate the size of the tumor. The data of each group were analyzed by t test. If the p value is less than 0.05, it is considered to be statistically significant.

Evaluation of effect: tumor inhibition rate = [1 - the average tumor weight of the treatment group / the weight of the average tumor of the control group] × 100%

Table 7 summarizes the efficacy of Form A of the compound of Formula I and the effect of free base on cancer inhibition.

The above experiments showed that the inhibition rate of Form A on human H1975 tumor xenograft inhibition was higher than that of free base.

Claims (38)

a crystalline form of a compound characterized by the aforementioned compound as shown in Formula I  The crystal form according to claim 1, wherein the X-ray powder diffraction spectrum of the crystal form has diffraction angles 2θ of about 4.8°, 7.5°, 13.2°, 19.0°, 19.6°, and 21.0°±0.2. Characteristic peak of °.    The crystal form according to claim 1 or 2, wherein the X-ray powder diffraction pattern of the crystal form has a diffraction angle 2θ of about 4.8°, 7.5°, 13.2°, 19.0°, 19.6°, and 21.0°. Characteristic peaks at 22.5° and 26.1° ± 0.2°.    The crystal form according to any one of claims 1 to 3, wherein the crystal form has a melting point of about 140.0 ° C to 145.0 ° C.    The crystal form according to any one of claims 1 to 4, wherein the crystal form has a melting point of about 142.0 °C.   The crystal form as described in any one of claims 1 to 5, wherein the crystal form has a purity 85%. The crystal form as described in any one of claims 1 to 6, wherein the crystal form has a purity 95%. The crystal form according to any one of claims 1 to 7, wherein the crystal form has a purity 99%. The crystal form as described in any one of claims 1 to 8, wherein the crystal form has a purity 99.5%.  The crystal form according to any one of claims 1 to 9, wherein the X-ray powder diffraction spectrum of the crystal form is as shown in FIG.    The crystal form according to the first aspect of the invention, wherein the X-ray powder diffraction spectrum of the crystal form has diffraction angles 2θ of about 6.4°, 17.9°, 19.1°, 20.0°, 21.5°, and 23.0°±0.2. Characteristic peak of °.    The crystal form according to claim 11, wherein the X-ray powder diffraction spectrum of the crystal form has diffraction angles 2θ of about 6.4°, 14.5°, 17.9°, 19.1°, 20.0°, 21.5°, 23.0. ° and characteristic peaks of 24.1 ° ± 0.2 °.    The crystal form as described in claim 11 or 12, wherein the X-ray powder diffraction pattern of the crystal form has a diffraction angle 2θ of about 6.4°, 11.8°, 14.5°, 15.9°, 17.9°, and 19.1°. Characteristic peaks of 20.0°, 21.5°, 23.0° and 24.1°±0.2°.    The crystal form according to any one of claims 11 to 13, wherein the crystal form has a melting point of about 133.0 ° C to 137.0 ° C.    The crystalline form according to any one of claims 11 to 14, wherein the crystalline form has a melting point of about 135.1 °C.   The crystal form according to any one of the items 11 to 15, wherein the crystal form has a purity 85%. The crystal form according to any one of the items 11 to 16, wherein the crystal form has a purity 95%. The crystal form according to any one of the items 11 to 17, wherein the crystal form has a purity 99%. The crystal form according to any one of the items 11 to 18, wherein the crystal form has a purity 99.9%.  The crystal form according to any one of the items 11 to 19, wherein the X-ray powder diffraction spectrum of the crystal form is as shown in Fig. 2 .   A process for the preparation of a crystalline form of a compound of formula I, characterized in that it comprises: a) N- (5-(5-chloro-4-(2-(2-(dimethylamino))-2-oxy) Ethylamino)phenylamino)pyrimidin-2-ylamino)-4-methoxy-2-(4-methylpiperazin-1-yl)phenyl)propenylamine and succinic acid are dissolved in In a solvent, the mixture is stirred at room temperature (RT) - 50 ° C for 8 to 96 hours, and the crystal form as described in any one of claims 2 to 10 is recovered; or b) N - (5- (5-Chloro-4-(2-(2-(dimethylamino)-2-oxoethyl)phenylamino)pyrimidin-2-ylamino)-4-methoxy-2- (4-Methylpiperazin-1-yl)phenyl)propenylamine and succinic acid are dissolved in a solvent, and the mixture is stirred at 50 ° C for 8 to 96 hours, and recovered as in claim 11 to 20 A crystal form as described.  The method of claim 21, wherein the solvent in the step a) is ethyl acetate or acetone.    The method of claim 21, wherein the solvent in the step b) is ethyl acetate.    A pharmaceutical composition characterized by comprising a therapeutically effective amount of a crystalline form and a pharmaceutically acceptable inactive component as set forth in any one of claims 1 to 20.    A pharmaceutical composition characterized by comprising a therapeutically effective amount of the crystalline form described in any one of claims 1 to 20 in combination with at least one other active ingredient.    The pharmaceutical composition according to claim 24, wherein the composition is administered orally.    The pharmaceutical composition according to claim 26, wherein the composition is a tablet or a capsule.    The pharmaceutical composition according to any one of claims 1 to 20, wherein the composition comprises 0.01% by weight to 99% by weight of the crystalline form described in any one of claims 1 to 20. .    The pharmaceutical composition according to claim 28, wherein the composition comprises 10% by weight to 50% by weight of the crystalline form described in any one of claims 1 to 20.    The pharmaceutical composition according to claim 29, wherein the composition comprises 10% by weight to 30% by weight of the crystalline form described in any one of claims 1 to 20.    A pharmaceutical composition according to any one of claims 1 to 20, or a pharmaceutical composition according to any one of claims 24 to 30, which is for use in the treatment of a disease for treating a subject The use of the drug.    The use according to claim 31, wherein the medicament is for treating, preventing, delaying or preventing the occurrence or progression of cancer, cancer metastasis, cardiovascular disease, immunological disease or ocular condition.    The use according to claim 32, wherein the drug is used as a kinase inhibitor.    The use according to claim 33, wherein the kinase comprises EGFR, ALK, ALK fusion protein, Flt3, Jak3, Blk, Bmx, Btk, HER2 (ErbB2), HER4 (ErbB4), Itk, Tec or Txk .    The use according to the invention of claim 34, wherein the EGFR is a mutated EGFR; the cancer is an EGFR-driven cancer, and the EGFR-driven cancer is characterized by one or more of the following mutations: (i) L858R, (ii) T790M, (iii) L858R and T790M, (iv) delE746_A750 or (v) delE746_A750 and T790M.    The use of the EGFR-driven cancer refers to non-small cell lung cancer, glioblastoma, pancreatic cancer, head and neck cancer, breast cancer, colon cancer, epithelial cancer, ovarian cancer, prostate, as described in claim 35. Cancer or adenocarcinoma.    The use according to claim 34, wherein the aforementioned ALK fusion protein is EML4-ALK or NPM-ALK kinase.    The use according to any one of claims 31 to 37, wherein the subject to be treated is a human.