CA3112496A1

CA3112496A1 - Salts of substituted pyrrolopyrimidine cdk inhibitor, crystal and use thereof

Info

Publication number: CA3112496A1
Application number: CA3112496A
Authority: CA
Inventors: Peng Gao; Yangping XU; Qingyi Meng; Fei Liu; Zhongying SUN; Keru ZHANG; Zhilin Chen; Jinfeng JIANG; Chenghui LU
Original assignee: Chia Tai Tianqing Pharmaceutical Group Co Ltd; Lianyungang Runzhong Pharmaceutical Co Ltd; Shouyao Holdings Beijing Co Ltd
Current assignee: Chia Tai Tianqing Pharmaceutical Group Co Ltd; Lianyungang Runzhong Pharmaceutical Co Ltd; Shouyao Holdings Beijing Co Ltd
Priority date: 2018-09-13
Filing date: 2019-09-12
Publication date: 2020-03-19
Also published as: AU2019338032A1; CN112888690B; CN112888690A; WO2020052627A1; JP2022500458A

Abstract

The present application belongs to the field of pharmaceutical chemistry and relates to salts of a substituted pyrrolopyrimidine CDK inhibitor, and particularly relates to a compound, 7-cyclopentyl-6-N,N-dimethylaminoformyl-N-(5-(1,2,3,6-4H-pyridine-4-yl)pyridine-2-yl)-7H-pyrrolo[2,3-d]pyrimidine-2-amine maleate (as represented by formula II), a crystal form of the compound represented by formula II, a preparation method therefor, and a pharmaceutical composition. The present application also relates to a use of the salts of the compound and the pharmaceutical composition thereof in preparing a drug for treating diseases related to the inhibitory effects of CDK4 and/or CDK6.

Description

SALTS OF SUBSTITUTED PYRROLOPYRIMIDINE CDK INHIBITOR, CRYSTAL AND
USE THEREOF
CROSS-REFERENCE TO RELATED APPLICATIONS
The present application claims benefit and priority to the Chinese Patent No.
201811068352.5 filed with the National Intellectual Property Administration, PRC on September 13, 2018, the disclosure of which is incorporated herein by reference in its entirety.
TECHNICAL FIELD
The present application belongs to the field of pharmaceutical chemistry, relates to salts and crystalline forms of a substituted pyrrolopyrimidine CDK inhibitor, a preparation method thereof and a pharmaceutical composition thereof, and relates to a use of the compounds and the pharmaceutical composition thereof in preparing a drug for treating and/or preventing a disease related to the inhibitory effects of CDK4 and/or CDK6.
BACKGROUND
W02017162215 discloses a compound, 7-cyclopentyl-6-N,N-dimethylcarbamoyl-N-(5-(1,2,3,6-4H-pyridin-4-yl)pyridin-2-y1)-7H-pyrrolo[2,3-Apyrimidin-2-amine (hereinafter referred to as a compound of formula I) as a CDK4/CDK6 inhibitor and a hydrochloride thereof.
HN \ \ 0 / N
------ N " 0 H
Formula I
It is generally desirable for a drug to have following excellent properties:
pharmaceutical activity, pharmacokinetics, bioavailability, hygroscopicity, melting point, stability, solubility, purity, easiness in preparation and the like, so that the requirements of the drug on production, storage, preparation and the like are satisfied. Studies have found that the compound of formula I and the hydrochloride thereof are highly hygroscopic, which leads to adverse effects on drug production, storage and preparation, such as obvious adverse effects on the fluidity, storage, even stability of the hygroscopic drug, which is difficult to satisfy the requirements of industrial production. The drug with higher melting point is more beneficial to preparing solid dosage forms, for example, stick punch can be easily avoided in the process of preparing tablets, thus having more advantages in drug processing, stability and the like. Therefore, there is a need to provide a drug substance with low hygroscopicity and high melting point.
BRIEF SUMMARY

Date Recue/Date Received 2021-03-11 In one aspect, the present application provides a maleate of a compound of formula I:

HN
/ N
N N
Formula I
=
The present application also provides a compound of formula II:
HN

N N /N¨
H
HOOC / COOH
\=
Formula II
In another aspect, the present application provides a crystalline form of the compound of formula II, wherein an X-ray powder diffraction pattern of the crystalline form using Cu-Ka radiation has diffraction peaks expressed by 20 values at 8.63, 10.50, 14.99, 17.29, 18.71 and 19.84 degree ( ).
In another aspect, the present application provides a method for preparing a crystalline foim of a compound of formula II, comprising precipitating the compound of formula II
from a solvent, wherein the solvent is selected from the group consisting of methanol, ethanol, isopropanol, N-methylpyrrolidone and dimethyl sulfoxide.
In another aspect, the present application provides a crystalline composition, wherein the crystalline form of the compound of formula II accounts for 50% or more, by weight of the crystalline composition.
In another aspect, the present application provides a pharmaceutical composition comprising a therapeutically effective amount of a maleate of the compound of formula I, the compound of formula II, the crystalline form of the compound of formula II, or the crystalline composition.
In still another aspect, the present application provides a method for treating and/or preventing a CDK4- and/or CDK6-mediated disease, comprising administering to a subject in need thereof a therapeutically effective amount of the maleate of the compound of formula I, the compound of formula II, the crystalline form of the compound of formula II, the crystalline composition, or the pharmaceutical composition.
In still another aspect, the present application provides a use of the maleate of the compound of formula I, the compound of formula II, the crystalline form of the compound of formula II, the crystalline composition or the pharmaceutical composition in the preparation of a medicament for treating and/or preventing a CDK4- and/or CDK6-mediated disease.

2 Date Recue/Date Received 2021-03-11 In yet another aspect, the present application provides a use of the maleate of the compound of formula I, the compound of formula II, the crystalline form of the compound of formula II, the crystalline composition or the pharmaceutical composition for treating and/or preventing a CDK4-and/or CDK6-mediated disease.
In yet another aspect, the present application provides the maleate of the compound of formula I, the compound of formula II, the crystalline form of the compound of formula II, the crystalline composition or the pharmaceutical composition for use in treating and/or preventing a CDK4-and/or CDK6-mediated disease.
BRIEF DESCRIPTION OF THE DRAWINGS
FIG. 1 is an XRPD pattern of a crystalline form of a compound of formula II
prepared in Example 2; and FIG. 2 is a DSC pattern of a crystalline form of a compound of formula II
prepared in Example 2.
SUMMARY
The present application provides a maleate of 7-cyclopenty1-6-N,N-dimethylcarbamoyl-N-(5-(1,2,3,6-4H-pyridin-4-yl)pyridin-2-y1)-7H-pyrrolo[2,3-Apyrimidin-2-amine (hereinafter referred to as a maleate of a compound of formula I) and a crystalline form thereof, and the compound has low hygroscopicity and high melting point, and excellent properties in at least one of pharmaceutical activity, pharmacokinetics, bioavailability, stability, solubility, purity, easiness in preparation and the like.
In one aspect, the present application provides a maleate of a compound of formula I.
In some embodiments, the molar ratio of the compound of formula Ito maleic acid in the maleate of the compound of formula I above is 1:0.5-2.
In some embodiments, the molar ratio of the compound of formula Ito maleic acid in the maleate of the compound of formula I above is 1:0.5, 1:1, 1:1.5 or 1:2.
In some embodiments, the molar ratio of the compound of formula Ito maleic acid in the maleate of the compound of formula I above is 1:0.5-1 or 1:1-2.
In some embodiments, the molar ratio of the compound of formula Ito maleic acid in the maleate of the compound of formula I above is 1:1.
In some embodiments, the maleate of the compound of formula I is in a crystalline form.
In some embodiments, the maleate of the compound of formula I is a compound of formula II.

3 Date Recue/Date Received 2021-03-11 HN

N N N N¨

H
HOOC COOH
Formula II
In another aspect, the present application provides a method for preparing a maleate of a compound of formula I, comprising reacting the compound of formula I with maleic acid.
In some embodiments, a method for preparing a maleate of a compound of formula I comprises reacting the compound of formula I with maleic acid in a solvent to form a maleate of the compound of formula I.
In some embodiments, a method for preparing a maleate of a compound of formula I comprises reacting the compound of formula I with maleic acid in a solvent to form a salt, and precipitating the salt from the solvent.
In some embodiments, the reaction is performed in a solvent, wherein the solvent is selected from the group consisting of methanol, ethanol, isopropanol, N-methylpyrrolidone and dimethyl sulfoxide; in some embodiments, the solvent is ethanol.
In another aspect, the present application provides a crystalline form of the compound of formula II, wherein an X-ray powder diffraction (XRPD) pattern of the crystalline form using Cu-Ka radiation has diffraction peaks expressed by 20 values at 8.63, 10.50, 14.99, 17.29, 18.71 and 19.84 degree (); in some embodiments, an X-ray powder diffraction pattern of the crystalline form using Cu-Ka radiation has diffraction peaks expressed by 20 values at 8.63, 10.50, 14.99, 16.14, 16.45, 17.29, 18.71, 19.84 and 20.93 degree ( ); in some embodiments, an X-ray powder diffraction pattern of the crystalline form using Cu-Ka radiation has diffraction peaks expressed by 20 values at 8.63, 10.50, 11.35, 12.44, 14.99, 16.14, 16.45, 17.29, 18.71, 19.84, 20.93, 21.74, 24.45, 27.30, 27.55 and 29.26 degree ( ); in some embodiments, an X-ray powder diffraction pattern of the crystalline form using Cu-Ka radiation has diffraction peaks expressed by 20 values at 8.63, 10.50, 11.35, 12.44, 14.15, 14.99, 16.14, 16.45, 17.29, 17.97, 18.71, 19.84, 20.56, 20.93, 21.74, 21.97, 22.86, 23.65, 24.45, 25.51, 26.15, 27.30, 27.55, 29.26, 30.96 and 31.45 degree ( ).
In some embodiments, in an X-ray powder diffraction pattern of the crystalline form of the compound of formula II disclosed herein using Cu-Ka radiation, the peak positions and relative intensities of diffraction peaks are shown in Table 1 below:
Table 1. Peak positions and relative intensities of diffraction peaks in the X-ray powder diffraction pattern of the crystalline form of the compound of formula II

4 Date Recue/Date Received 2021-03-11 Relative 20 Relative Number Intensity (I/I0) Number Intensity (I/I0) (degree) (degree) 1 8.63 48.2 14 20.93 32.4 2 10.50 40.9 15 21.74 30.0 3 11.35 22.9 16 21.97 26.6 4 12.44 28.7 17 22.86 9.5

5 14.15 8.5 18 23.65 10.3

6 14.99 48.8 19 24.45 25.2

7 16.14 35.9 20 25.51 19.0

8 16.45 39.0 21 26.15 18.8

9 17.29 39.4 22 27.30 22.2

10 17.97 17.3 23 27.55 27.5

11 18.71 100.0 24 29.26 23.3

12 19.84 88.4 25 30.96 9.8

13 20.56 15.5 26 31.45 13.8 In some embodiments, the XRPD pattern of the crystalline form of the compound of formula II
provided herein is shown in FIG. 1.
Without limitation, the Differential Scanning Calorimetry (DSC) of the crystalline form of the compound of formula II provided herein shows an absorption peak at 237.67 C;
specifically, the Differential Scanning Calorimetry (DSC) pattern of the crystalline form of the compound of formula II
is shown in FIG. 2.
In the present application, the instrument for X-ray powder diffraction spectrometry is Bruker D8 X-ray diffractometer, and the conditions and methods are: copper target, 40 kv 40 mA; slit: 0.681 mm/5.5 mm; scan range: 5-40 , time [s]: 0.1; and step length: 0.02 .
In the present application, the instrument for DSC spectroscopy is METTLER
TOLEDO DSC1, and the condition and method is: heating at 10 C/min in 30-250 C.
In the present application, the melting point is measured with reference to the first method of melting point measurement of Chapter 0612 Chinese Pharmacopoeia (2015 Edition).
In the present application, the instrument for elemental analysis is Vario MICRO elemental analyzer, and the method is: JY/T 017-1996 General rules for elemental analyzer.
For any given crystalline form, the relative intensities of diffraction peaks may vary due to preferred orientations resulting from, e.g., crystal morphology, as is well known in the field of crystallography. The peak intensity varies at a place where there is preferred orientation effect, while it is impossible for the diffraction peak position of crystalline form to vary. In addition, there may be slight errors in the peak positions for any given crystalline form, as is also well known in the field of crystallography. For example, the peak positions may shift due to temperature changes, sample movement or calibration of the instrument when analyzing a sample, and the error in the measurement of 20 is sometimes about 0.2 degree, and therefore, it is well known to those skilled Date Recue/Date Received 2021-03-11 in the art that this error should be taken into account when determining each crystalline structure.
DSC measures the transition temperature when a crystalline form absorbs or releases heat due to a change in crystalline structure or melting of the crystalline form. For the same crystalline form of the same compound, the thermal transition temperature and melting point errors in continuous analyses are typically within about 5 C, usually within about 3 C, and a compound having a given DSC peak means that the DSC peak is 5 C. DSC provides an auxiliary method to identify different crystalline forms. Different crystal morphologies can be identified by their different transition temperature characteristics. It is noted that for a mixture, its DSC peak may vary over a larger range. Furthermore, melting temperature is related to heating rate due to the decomposition of a substance in the melting process.
In another aspect, the present application provides a method for preparing a crystalline form of a compound of formula II, comprising precipitating the compound of formula II
from a solvent.
In some embodiments, a method for preparing a crystalline form of a compound of formula II
comprises precipitating the compound of formula II from a solvent, wherein the solvent is selected from the group consisting of methanol, ethanol, isopropanol, N-methylpyrrolidone and dimethyl sulfoxide. In some embodiments, the solvent is ethanol.
In some embodiments, a method for preparing a crystalline form of a compound of formula II
comprises the steps of:
HN-------.., HN
N N-----) ______________________ e _.,..._ _......
N N N a N¨ N N N N¨

H , _ H
HOOC\_, COOH a /
Formula I Formula II
1) reacting the compound of formula I with maleic acid in a solvent; and 2) precipitating a solid.
In some embodiments, step 1) is: mixing maleic acid with a solvent, mixing a compound of formula I with a solvent, and mixing the two mixtures and performing reaction.
In some embodiments, the molar ratio of the maleic acid to the compound of formula I in step 1) is 1:1-1.2.
In some embodiments, the solvent is selected from the group consisting of methanol, ethanol, isopropanol, N-methylpyrrolidone and dimethyl sulfoxide; in some embodiments, the solvent is ethanol.
In some embodiments, the reaction in step 1) is performed under a heating condition; in some embodiments, the heating temperature is 65-75 C.
In some embodiments, step 1) is: mixing maleic acid with a solvent, mixing a compound of the formula I with a solvent, mixing the two mixtures and performing reaction under a heating Date Recue/Date Received 2021-03-11 condition, and then reducing the temperature.
In some embodiments, the temperature is reduced to 20-30 C; in some specific embodiments, the temperature is reduced to 20-30 C and then the mixtures are stirred, for example, stirred for 1 h.
In some embodiments, the step 2) is followed by separating the solid precipitated in step 2); in some specific embodiments, the method further comprises drying the separated solid, for example, drying at 40-65 C under reduced pressure.
In still another aspect, the present application provides a crystalline composition comprising a crystalline form of a compound of formula II, wherein the crystalline form of the compound of formula II accounts for 50% or more, preferably 80% or more, more preferably 90% or more, and most preferably 95% or more, by weight of the crystalline composition.
In still another aspect, the present application provides a pharmaceutical composition comprising a therapeutically effective amount of a maleate of a compound of formula I, a compound of formula II, a crystalline form of a compound of formula II, or a crystalline composition of the crystalline form of the compound of formula II described herein. The pharmaceutical composition disclosed herein may or may not contain a pharmaceutically acceptable excipient. In addition, the pharmaceutical composition disclosed herein may further comprise one or more other therapeutic agents.
In still another aspect, the present application provides a method for treating and/or preventing a CDK4- and/or CDK6-mediated disease, comprising administering to a subject in need thereof a therapeutically effective amount of a maleate of a compound of formula I, a compound of formula II, a crystalline form of a compound of formula II, a crystalline composition of the crystalline form of the compound of formula II, or a pharmaceutical composition thereof.
In still another aspect, the present application provides a use of a maleate of a compound of formula I, a compound of formula II, a crystalline form of the compound of formula II, a crystalline form composition of the crystalline form of the compound of formula II, or a pharmaceutical composition thereof in the preparation of a medicament for treating and/or preventing a CDK4- and/or CDK6-mediated disease.
In still another aspect, the present application provides a use of a maleate of a compound of formula I, a compound of formula II, a crystalline form of the compound of formula II, a crystalline composition of the crystalline form of the compound of formula II, or a pharmaceutical composition thereof for treating and/or preventing a CDK4- and/or CDK6-mediated disease.
In still another aspect, the present application provides a maleate of a compound of formula I, a compound of formula II, a crystalline form of the compound of formula II, a crystalline composition of the compound of formula II, or a pharmaceutical composition thereof for use in treating and/or preventing a CDK4- and/or CDK6-mediated disease.

Date Recue/Date Received 2021-03-11 In some embodiments of the present application, the CDK4- and/or CDK6-mediated disease comprises cancer.
In some embodiments of the present application, the cancer includes, but is not limited to: bladder cancer; breast cancer, such as metastatic breast cancer; colon cancer; kidney cancer; epidermoid carcinoma; liver cancer; lung cancer, such as small cell lung cancer and non-small cell lung cancer;
esophageal cancer; gallbladder cancer; ovarian cancer; pancreatic cancer, such as exocrine pancreatic cancer; stomach cancer; cervical cancer; thyroid cancer; nasal cancer; head and neck cancer; prostate cancer; skin cancer, such as squamous cell carcinoma;
lymphoid hematopoietic tumor, such as leukemia, acute lymphocytic leukemia, chronic lymphocytic leukemia, B-cell lymphoma, T-cell lymphoma, multiple myeloma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, hairy cell lymphoma and Burkitt's lymphoma; myeloid hematopoietic tumor, such as acute and chronic myeloid leukemia, myelodysplastic syndrome and promyelocytic leukemia;
thyroid follicular cancer; tumors from stromal cells, such as fibrosarcoma and rhabdomyosarcoma; tumors of the central or peripheral nervous system, such as astrocytoma, neuroblastoma, neuroglioma and schwannoma; melanoma; seminoma; teratoma; osteosarcoma; xeroderma pigmentosum;

keratoacanthoma; follicular thyroid carcinoma; liposarcoma; neuroendocrine tumors; and Kaposi's sarcoma.
The "pharmaceutically acceptable excipient" refers to an inert substance administered with active ingredient to facilitate administration of the active ingredient, including, but not limited to, any glidant, sweetener, diluent, preservative, dye/colorant, flavor enhancer, surfactant, wetting agent, dispersant, disintegrant, suspending agent, stabilizer, isotonizing agent, solvent or emulsifier acceptable for use in humans or animals (e.g., domesticated animals) as permitted by the National Medical Products Administration. Non-limiting examples of the excipients include calcium carbonate, calcium phosphate, various sugars and types of starch, cellulose derivatives, gelatin, vegetable oils, and polyethylene glycols.
The pharmaceutical composition disclosed herein may be formulated into solid, semi-solid, liquid or gaseous formulations such as tablets, pills, capsules, powders, granules, ointments, emulsions, suspensions, solutions, suppositories, injections, inhalants, gels, microspheres, and aerosols. The pharmaceutical composition disclosed herein may be prepared by methods known in the art, such as by conventional mixing, dissolving, granulating, dragee-making, levigating, emulsifying, and lyophilizing.
Typical routes of administration of the pharmaceutical composition disclosed herein include, but are not limited to, oral, rectal, transmucosal, enteral administration, or topical, percutaneous, inhalational, parenteral, sublingual, intravaginal, intranasal, intraocular, intraperitoneal, intramuscular, subcutaneous and intravenous administration. The preferred route of administration Date Recue/Date Received 2021-03-11 is oral administration.
The term "treating" means administering the compound or formulation described herein to ameliorate or eliminate a disease or one or more symptoms associated with the disease, and includes:
(i) inhibiting a disease or disease state, i.e., arresting its development;
(ii) alleviating a disease or disease state, i.e., causing its regression.
The term "preventing" means administering the compound or formulation described herein to prevent a disease or one or more symptoms associated with the disease, and includes: preventing the occurrence of the disease or disease state in a mammal, particularly when such a mammal is predisposed to the disease state but has not yet been diagnosed as having it.
For drugs and pharmacological active agents, the term "therapeutically effective amount" refers to an amount of a drug or a medicament that is sufficient to provide the desired effect but is non-toxic.
The determination of the effective amount varies from person to person, depending on the age and general condition of a subject and also depending on the particular active substance. The appropriate effective amount in a case may be determined by those skilled in the art in the light of routine tests.
A therapeutically effective amount of the maleate of the compound of formula I, the compound of formula II or a crystalline form thereof is from about 0.0001 to 20 mg/kg bw/day, for example from 0.001 to 10 mg/kg bw/day.
The dose frequency of the maleate of the compound of formula I, the compound of formula II or a crystalline form thereof depends on needs of an individual patient, e.g., once or twice daily or more times daily. Administration may be intermittent, for example, in a period of several days, a patient receives a daily dose of the compound of formula I, the compound of formula II
or a crystalline form thereof, and in the following period of the several days or more days, the patient does not receive the daily dose of the compound of formula I, the compound of formula II or a crystalline form thereof.
EXAMPLES
The following specific examples are presented to enable those skilled in the art to more clearly understand and practice the present application. These specific examples should not be considered as limiting the scope of the present application, but merely as being exemplary description and representative of the present application. It should be understood by those skilled in the art that there are other synthesis routes to the compounds of the present application, and the following non-limiting examples are provided. Unless otherwise stated, all starting materials used in the present application were commercially available and used without further purification.

Date Recue/Date Received 2021-03-11 Example 1: Preparation of a Compound of Formula I
H N
H N
N N
N N
N N N N¨

H N¨

H /
Formula 1-1 Formula 1 _ The product obtained by the preparation method with reference to Example 6 of was dissolved in heated ethanol, the mixture was cooled to precipitate a solid and filtered, and the filter cake was dried under reduced pressure to give a crystalline form of a compound of formula I-1 (melting point < 210 C).
The above compound of formula I-1 (2.26 kg) was added into anhydrous methanol (22.6 L); the mixture was stirred at 10-30 C to dissolve, added dropwise with 2 mol/L
sodium hydroxide aqueous solution to adjust the pH to 9-10, then stirred for more than 1 h after dropwise adding, and filtered; the filter cake was washed by purified water until the pH of a washing liquid was about 7;
and the wet product was dried at 45-55 C under reduced pressure for more than 8 h until the weight was constant to give a crystalline form (1.71 kg) of the compound of formula I. HRMS (ESI+, [M+Hr) m/z 432.2511, melting point: 215-217 C.
Example 2: Preparation of a Compound of Formula II
H N
H N
N N N N
N N N N¨ _______________________________________________________ H
HOOC\_, COOH
Formula 1 Formula II
Absolute ethanol (25.5 L) and a compound (1.7 Kg) of formula I were added into a reaction flask, stirred for 10 min, and heated under nitrogen atmosphere; the reaction mixture was controlled at 65-75 C, stirred for 10 min, added with maleic acid (503 g) and absolute ethanol solution (1.7 L), continuously kept at 65-75 C, stirred for 5 h, cooled, controlled at 20-30 C, stirred for 1 h and filtered; the filter cake was rinsed with absolute ethanol (3.4 L); the wet product was dried at 60 5 C under reduced pressure for 12 h to give a crystalline form of the compound of formula II, wherein the melting point is above 235 C, the X-ray powder diffraction pattern of the crystalline form using Cu-Ka ray is shown in FIG. 1, the differential scanning calorimetry pattern of the crystalline form is shown in FIG. 2, and the elemental analysis results thereof are as follows:
calculated value (%): 61.41% C atoms, 6.07% H atoms and 17.90% N atoms, and measurement value: 61.13% C atoms, 6.08% H atoms and 17.99% N atoms.
Experimental Example 1: Hygroscopicity Date Recue/Date Received 2021-03-11 The experiment comprises the following steps: about 30 mg of samples (the crystalline form of the compound of formula I in Example 1, the crystalline form of the compound of formula II in Example 2, and the crystalline form of the compound of formula I-1 in Example 1) were measured in a Dynamic Vapor Sorption (DVS) sample tray.
Instrument model: SMS DVS Advantage Experimental condition: 25 C
Balance dm/dt: 0.002 %/min: (time: 10 min-max 360 min) RH (%) measurement gradient: 10 %
RH (%) measurement gradient range: 0%-90%-0%
The results are shown in Table 2 below.
Table 2 Experimental Compound of Compound of Compound of condition formula I formula II formula I-1 Relative humidity RH (%) Moisture gain (%) 30% 7.78 0.11 6.61 60% 8.15 0.18 11.63 80% 8.30 0.23 16.35 Experimental Example 2: Stability A sample of the crystalline form of the compound of formula II in Example 2 was placed under experimental conditions of influencing factors (40 C incubator, 60 C
incubator, and high humidity environment of 75% RH and 92.5% RH). The sample was taken and detected at 5, 10 and 30 days.
Instrument: Waters e2695 high performance liquid chromatography;
Detector: Waters 2489 UV absorption detector/2998 diode array detector;
Chromatographic column: Phenomenex Titank C18 (4.6x150 mm, 3 pm);
Injection volume: 10 jiL; Detection wavelength: 279 nm; Flow rate: 1.1 mL/min;
Column temperature: 35 C;
Mobile phases: phase A: 30 mmol/L ammonium formate solution (pH was adjusted to 7.5 with ammonium hydroxide), phase B: acetonitrile.
Preparation of sample solution: appropriate amounts of each sample in the above experimental conditions were weighed out precisely, and dissolved in a solvent (30%
acetonitrile aqueous solution) to give a solution containing about 1.0 mg of the compound of formula II per 1 mL, as the concentration of relevant substance for testing. The results are shown in Table 3.
Table 3 Content of compound of Experimental condition formula II (%) Day 0 99.5%

Date Recue/Date Received 2021-03-11 Content of compound of Experimental condition formula II (%) 40 C_5 days 99.4%
40 C_10 days 99.4%
40 C_30 days 99.3%
60 C_5 days 99.5%
60 C_10 days 99.4%
60 C_30 days 99.2%
75% RHat room _ 99.5%
temperature for 5 days 75% RH_at room 99.5%
temperature for 10 days 75% RH_at room 99.4%
temperature for 30 days 92.5% RH_at room 99.5%
temperature for 5 days 92.5% RH_at room 99.5%
temperature for 10 days 92.5% RH_at room 99.4%
temperature for 30 days Experimental Example 3: Measurement of Proliferation Inhibitory Effect of the Compound on Human Colon Cancer Cell Line Colo-205 A number of Colo-205 cells in logarithmic growth phase were seeded in a 96-well plate. After 24 h of growth, DMSO solutions with different concentrations (1, 3, 10, 30, 100, 300, 1000, 3000, 10000 nM) of the compound of formula II in Example 2 were added and incubated for 6 days. After the reaction of the compound of formula II was completed, MTT working solution was added into each well; after 4 h, the triple solution (10% Sodium dodecyl sulfate, 5%
isobutanol, 0.012 mol/L HC1) was dissolved, OD value was measured at 570 nm wavelength by an microplate reader (Synergy H4); and the cell growth inhibition rate was calculated by the following formula: inhibition = (OD
value control well - OD value dosing well) / OD value control well x100%.
Median Inhibitory Concentration IC50 was calculated from concentration inhibition rate using GraphPad Prism 7 software by a non-linear regression method. The results are shown in Table 4.
Table 4 Compound IC50(Colo-205) / nM
Compound of 301.0 formula II

Date Recue/Date Received 2021-03-11

Claims

CA 03112496 2021-03-11

1. A maleate of a compound of formula I

HN õ, \
/ N
I N
Formula I
optionally, the molar ratio of the compound of formula I to maleic acid is 1:0.5-2.

2. The maleate of the compound of formula I according to claim 1, wherein the maleate of the compound of formula I is the compound of formula II
HN
NNN N¨

H
HOOC / COOH
\= /
Formula II

3. A crystalline form of a compound of formula II
HN
N /P
N N¨

H
HOOC COOH
Formula II
wherein an X-ray powder diffraction pattern of the crystalline form using Cu-Ka radiation has diffraction peaks expressed by 20 values at 8.63, 10.50, 14.99, 17.29, 18.71 and 19.84 degree ( );
typically, has diffraction peaks expressed by 20 values at 8.63, 10.50, 14.99, 16.14, 16.45, 17.29, 18.71, 19.84 and 20.93 degree ( ); typically, has diffraction peaks expressed by 20 values at 8.63, 10.50, 11.35, 12.44, 14.99, 16.14, 16.45, 17.29, 18.71, 19.84, 20.93, 21.74, 24.45, 27.30, 27.55 and 29.26 degree ( ); and typically, has diffraction peaks expressed by 20 values at 8.63, 10.50, 11.35, 12.44, 14.15, 14.99, 16.14, 16.45, 17.29, 17.97, 18.71, 19.84, 20.56, 20.93, 21.74, 21.97, 22.86, 23.65, 24.45, 25.51, 26.15, 27.30, 27.55, 29.26, 30.96 and 31.45 degree ( ).

4. The crystalline fonn of the compound of formula II according to claim 3, wherein in the X-ray powder diffraction pattern of the crystalline form using Cu-Ka radiation, the peak positions and relative intensities of diffraction peaks are shown in the table below:

Date Recue/Date Received 2021-03-11 Relative 20 Relative Number Intensity (I/Io) Number Intensity (I/Io) (degree) (degree) (%) (%) 1 8.63 48.2 14 20.93 32.4 2 10.50 40.9 15 21.74 30.0 3 11.35 22.9 16 21.97 26.6 4 12.44 28.7 17 22.86 9.5 5 14.15 8.5 18 23.65 10.3 6 14.99 48.8 19 24.45 25.2 7 16.14 35.9 20 25.51 19.0 8 16.45 39.0 21 26.15 18.8 9 17.29 39.4 22 27.30 22.2 10 17.97 17.3 23 27.55 27.5 11 18.71 100.0 24 29.26 23.3 12 19.84 88.4 25 30.96 9.8 13 20.56 15.5 26 31.45 13.8

5. The crystalline form of the compound of formula II according to claim 3, wherein the X-ray powder diffraction pattern of the crystalline form is shown in FIG. 1.

6. A crystalline form of a compound of formula II
HNI
N N----) ______________________________________ /P
NNN N-H
HOOC
\=,COOH a, Formula II , wherein a differential scanning calorimetry thereof shows an absorption peak at 237.67 C.

7. The crystalline form of the compound of formula II according to claim 6, wherein a differential scanning calorimetry pattern of the crystalline form is shown in FIG. 2.

8. A method for preparing the crystalline form of the compound of formula II
according to any one of claims 3 to 7, comprising precipitating the compound of formula II from a solvent, wherein the solvent is selected from the group consisting of methanol, ethanol, isopropanol, N-methylpyrrolidone and dimethyl sulfoxide; typically, wherein the solvent is ethanol.

9. A crystalline composition, wherein the crystalline foim of the compound of formula II
according to any one of claims 3 to 7 accounts for 50% or more, 80% or more, 90% or more, or 95% or more, by weight of the crystalline composition.

10. A pharmaceutical composition, comprising a therapeutically effective amount of the maleate of the compound of formula I according to claim 1 or 2, the crystalline form of the compound of formula II according to any one of claims 3 to 7, or the crystalline composition according to claim 9.

11. A method for treating a CDK4- and/or CDK6-mediated disease, comprising administering to a Date Recue/Date Received 2021-03-11 subject in need thereof a therapeutically effective amount of the maleate of the compound of formula I according to claim 1 or 2, the crystalline form of the compound of formula II
according to any one of claims 3 to 7, the crystalline composition according to claim 9, or the pharmaceutical composition according to claim 10.

12. A use of the maleate of the compound of formula I according to claim 1 or 2, the crystalline form of the compound of formula II according to any one of claims 3 to 7, the crystalline composition according to claim 9, or the pharmaceutical composition according to claim 10 in the preparation of a medicament for treating and/or preventing a CDK4- and/or CDK6-mediated disease.

13. A use of the maleate of the compound of formula I according to claim 1 or 2, the crystalline form of the compound of formula II according to any one of claims 3 to 7, the crystalline composition according to claim 9, or the pharmaceutical composition according to claim 10 in treating and/or preventing a CDK4- and/or CDK6-mediated disease.

14. The maleate of the compound of formula I according to claim 1 or 2, the crystalline form of the compound of formula II according to any one of claims 3 to 7, the crystalline composition according to claim 9, or the pharmaceutical composition according to claim 10 for use in treating and/or preventing a CDK4- and/or CDK6-mediated disease.

15. The method according to claim 11, the use according to claim 12 or 13, or the maleate of the compound of formula I, the crystalline form of the compound of formula II, the crystalline composition or the pharmaceutical composition according to claim 14, wherein the CDK4-and/or CDK6-mediated disease is cancer; typically, the cancer is selected from the group consisting of bladder cancer, breast cancer, colon cancer, renal cancer, epidermoid carcinoma, liver cancer, lung cancer, esophageal cancer, gallbladder cancer, ovarian cancer, pancreatic cancer, stomach cancer, cervical cancer, thyroid cancer, nasal cancer, head and neck cancer, prostate cancer, skin cancer, lymphoid hematopoietic tumor, myeloid hematopoietic tumor, follicular thyroid cancer, tumors from stromal cells, tumors of the central or peripheral nervous system, melanoma, seminoma, teratoma, osteosarcoma, xeroderma pigmentosum, keratoacanthoma, follicular thyroid carcinoma, liposarcoma, neuroendocrine tumors, and Kaposi's sarcoma.
Date Recue/Date Received 2021-03-11