TWI745289B - A crystalline form of cyclin-dependent protein kinase inhibitor and preparation methods thereof - Google Patents

Abstract

The invention relates to a crystalline of a cyclin-dependent protein kinase inhibitor and preparation methods thereof. Specifically, the invention relates to form II crystal of cyclin-dependent protein kinase inhibitor (CDK4&6) and preparation methods thereof. More specifically, the present invention relates to form II crystal of 6-acetyl-8-cyclopentyl-5-methyl-2-((5-(piperidin-4-yl)pyridin-2-yl)amino)pyrido[2,3-d]pyrimidin-7(8H)-one (compound of formula (I)) and preparation methods thereof. The form II crystal in this invention has X-ray powder diffraction pattern as shown in Figure 1. The form II crystal in this invention has good polymorph stability and chemical stability, furthermore, the solvent used for crystallization is low toxicity and low residual, so it can be better used for clinical treatment.

Description

Crystal form of cyclin-dependent protein kinase inhibitor and preparation method thereof

The present invention relates to 6-acetyl-8-cyclopentyl-5-methyl-2-((5-(piperidin-4-yl)pyridin-2-yl)amino)pyrido[2,3- d] Pyrimidine-7(8H)-one and its II crystal form. The compound of formula (I) prepared according to the method of the present invention can be used for the treatment of breast cancer.

Breast cancer is one of the most common malignant tumors in women. It has a high incidence and is quite aggressive, but the course of the disease is slow. On February 1, 2010, the China Population Association released the "China Breast Disease Investigation Report" in Beijing. The report showed that, The mortality rate of breast cancer in urban areas in my country has increased by 38.91%. Breast cancer has become the disease that poses the greatest threat to women’s health. There are currently at least 156 breast cancer drugs being developed and marketed, of which 68% are targeted therapy drugs. It is found that tumors are related to abnormal cell cycle. A large number of mutations in mitotic signaling proteins and defects in anti-mitotic signaling proteins in tumor cells lead to proliferation disorders. At the same time, most tumors have genomic instability (GIN) and chromosome instability (CIN). These three basic cell cycle defects are Directly or indirectly caused by the loss of control of CDKs. Cyclin Dependent Kinase (CDK, Cyclin Dependent Kinase) inhibitors are increasingly becoming popular targets.

At present, there are many first- and second-generation CDK inhibitors developed. The most concerned second-generation drugs include the CDK4&6 inhibitor PD-0332991 jointly developed by Pfizer and Onyx. It inhibits the activity of CDK4&6, inhibits the phosphorylation of Rb, and makes The E2F-Rb complex stays in the cytoplasm and blocks the initiation of the cell cycle. Clinical trial results (NCT00721409) showed that the progression-free survival (PFS) of patients treated with letrozole monotherapy was 7.5 months, while patients treated with letrozole and PD-0332991 had no progression. The survival period was extended to 26.1 months. This significant advantage has received widespread attention. In early 2013, the FDA reviewed the interim results of this drug and believed that it may be a breakthrough anti-cancer drug.

WO2014183520 discloses a CDK4&6 inhibitor similar in structure to PD-0332991, which has significant CDK4&6 inhibitory activity and high selectivity, and includes the following compounds:

However, WO2014183520 did not thoroughly study the crystalline form of the compound. It is well known to those skilled in the art that the crystal structure of the active ingredient used in medicine often affects the chemical stability of the drug. The difference in crystallization conditions and storage conditions may lead to changes in the crystal structure of the compound, sometimes accompanied by Other crystal forms. Generally speaking, there are no rules for amorphous drug products Its crystal structure often has other defects, such as poor product stability, finer crystallization, difficult filtration, easy agglomeration, and poor fluidity. Therefore, it is necessary to improve the various properties of the above-mentioned compounds. We need in-depth research to find new crystal forms with higher crystal purity and good chemical stability.

The invention provides 6-acetyl-8-cyclopentyl-5-methyl-2-((5-(piperidin-4-yl)pyridin-2-yl)amino)pyrido[2,3 -d] A new crystalline form of pyrimidine-7(8H)-one (as shown in formula (I)).

A series of crystalline products of the compound represented by formula (I) obtained under different crystallization conditions. The obtained crystalline products were tested by X-ray diffraction and DSC. It was found that the compound represented by formula (I) can obtain a The crystal form with good stability, we call it type II crystal. The DSC spectrum of the type II crystal in this application shows that there is a melting endothermic peak near 294.42°C. The X-ray powder diffraction pattern is shown in Figure 1, using Cu-Ka radiation, with a 2 θ angle and interplanar spacing (d value). ) Represents the X-ray powder diffraction pattern, in which 5.84 (15.12), 6.16 (14.35), 10.75 (8.22), 12.56 (7.04), 12.96 (6.82), 16.25 (5.45), 17.24 (5.14), 18.97 (4.67) ), 20.22 (4.39), 21.74 (4.08), 22.99 (3.87), and 25.85 (3.44) have characteristic peaks.

The invention also provides the preparation of 6-acetyl-8-cyclopentyl-5-methyl-2-((5-(piperidin-4-yl)pyridin-2-yl)amino)pyrido[2 ,3-d)pyrimidine-7(8H)- A method for the type II crystallization of ketones, the method includes the following steps: 1) Add any crystalline or amorphous compound of formula (I) to an appropriate amount of solvent, add an acid, and dissolve it until it is clear, then add a base, and precipitate Crystal, the solvent is selected from any one of alcohols, ketones, and nitriles with carbon atoms less than or equal to 3 or a mixed solvent of them and water; 2) filtering the crystals, washing, and drying.

In a preferred embodiment, the acid is an inorganic acid, preferably hydrochloric acid; the base is an inorganic base, preferably sodium hydroxide or potassium hydroxide.

In a preferred embodiment, the solvent described in step 1) is methanol, ethanol, isopropanol, acetone, acetonitrile or methanol/water, ethanol/water, acetone/water, acetonitrile/water, isopropanol/water; Preferably it is ethanol.

The method of recrystallization is not particularly limited, and it can be performed by a usual recrystallization operation method. For example, the compound represented by formula (I) can be used as the raw material to dissolve in an organic solvent by heating and slowly cooling to crystallize. After the crystallization is completed, it can be filtered and dried to obtain the desired crystal. It should be noted that the filtered crystals are usually vacuum-dried under reduced pressure at about 30 to 100°C, preferably 40 to 60°C, to achieve the effect of removing the recrystallization solvent.

By means of differential scanning calorimetry (DSC) and X-diffraction pattern measurement, the obtained crystals of the compound represented by formula (I) were studied for the crystal structure, and the solvent residues of the obtained crystals were detected at the same time.

The type II crystals of compound represented by formula (I) prepared according to the method of the present invention do not contain or only contain a relatively low content of residual solvents, and meet the limit requirements of the relevant pharmaceutical product residual solvents stipulated by the National Pharmacopoeia. Therefore, the crystals of the present invention It can be used as an active ingredient in medicine.

Studies have shown that the type II crystals of the compound represented by formula (I) prepared by the present invention have good stability under conditions of light, high temperature and high humidity, and have good crystal stability under conditions such as grinding, pressure and heat. It can meet the medicinal requirements of production, transportation and storage, the production process is stable, repeatable and controllable, and can be adapted to industrial production.

Figure 1 is an X-ray powder diffraction pattern of the compound II type crystal represented by formula (I).

Figure 2 is the DSC spectrum of the compound II crystal of formula (I).

Hereinafter, the present invention will be explained in more detail in conjunction with the embodiments. The embodiments of the present invention are only used to illustrate the technical solutions of the present invention, and do not limit the essence and scope of the present invention.

Test equipment used in the experiment

1. DSC spectrum

Instrument model: MettlerToledo DSC 1 Staree System

Purge gas: nitrogen

Heating rate: 10.0℃/min

Temperature range: 40-350℃

2. X-ray diffraction spectrum

Instrument model: Bruker D8 Focus X-ray powder diffractometer

Ray: Monochromatic Cu-K α rays (λ=1.5406)

Scanning method: θ/2 θ, scanning range: 2-40°

Voltage: 40KV, current: 40mA

Example 1

Take (1.0g, 2.24mmol) of the compound represented by formula (I) (prepared according to the method provided by WO2014183520) into a 250ml Erlenmeyer flask, add 40ml of ethanol, stir at room temperature, and then drop in dilute hydrochloric acid (219mg, 6.01mmol ) (Dissolved in 4ml water), heated to 60°C, dissolved until clear, dropped into sodium hydroxide solution (576mg, 14.40mmol) (dissolved in 40ml water), cooled to room temperature and stirred overnight. After drying, 0.88 g of solid was obtained, and the yield was 88.0%. The X-ray diffraction spectrum of the crystalline sample is shown in Figure 1. The crystals are about 5.84 (15.12), 6.16 (14.35), 10.75 (8.22), 12.56 (7.04), 12.96 (6.82), 16.25 (5.45), 17.24 (5.14), 18.97 (4.67), 20.22 (4.39), 21.74 (4.08), 22.99 (3.87), and 25.85 (3.44) have characteristic peaks. The DSC spectrum is shown in Figure 2, with a sharp melting endothermic peak at 294.42°C. This crystal form is defined as the II crystal form.

Example 2

Take (1.0g, 2.24mmol) of the compound represented by formula (I) (prepared according to Example 1) into a 250ml Erlenmeyer flask, add 40ml of methanol, stir at room temperature, and then drop in dilute hydrochloric acid (219mg, 6.01mmol) (Dissolve in 4ml water), heat to 60℃, dissolve until clear, add dropwise to sodium hydroxide solution (576g, 14.40mmol) (dissolved in 40ml water), cool to room temperature and stir overnight. After drying, 0.86 g of solid was obtained, and the yield was 86.0%. The X-diffraction and DSC spectra were studied and compared, and the product was determined to be of crystalline form II.

Example 3

Take (1.0g, 2.24mmol) of the compound represented by formula (I) (prepared according to Example 1) into a 250ml Erlenmeyer flask, add 40ml of isopropanol, and stir at room temperature. Then drop in dilute hydrochloric acid (219mg, 6.01mmol) (dissolved in 4ml of water), heat to 60℃, dissolve until clear, drop into sodium hydroxide solution (576mg, 14.40mmol) (dissolved in 40ml of water), and cool to room temperature Warm stirring overnight. After drying, 0.90 g of solid was obtained, and the yield was 90.0%. The X-diffraction and DSC spectra were studied and compared, and the product was determined to be of crystalline form II.

Example 4

Take (1.0g, 2.24mmol) of the compound represented by formula (I) (prepared according to Example 1) into a 250ml Erlenmeyer flask, add 40ml of acetone, stir at room temperature, and then drop in dilute hydrochloric acid (219mg, 6.01mmol) (Dissolve in 4ml water), heat to 60°C, dissolve until clear, drop into sodium hydroxide solution (576mg, 14.40mmol) (dissolved in 40ml water), cool to room temperature and stir overnight. After drying, 0.86 g of solid was obtained, and the yield was 86.0%. The X-diffraction and DSC spectra were studied and compared, and the product was determined to be of crystalline form II.

Example 5

Take (1.0g, 2.24mmol) of the compound represented by formula (I) (prepared according to Example 1) into a 250ml Erlenmeyer flask, add 40ml of acetonitrile, stir at room temperature, and then drop in dilute hydrochloric acid (219mg, 6.01mmol) (Dissolve in 4ml water), heat to 60°C, dissolve until clear, drop into sodium hydroxide solution (576mg, 14.40mmol) (dissolved in 40ml water), cool to room temperature and stir overnight. 0.60 g of solid was obtained after drying, and the yield was 60.0%. The X-diffraction and DSC spectra were studied and compared, and the product was determined to be of crystalline form II.

Example 6

Take (1.0g, 2.24mmol) of the compound represented by formula (I) (prepared according to Example 1) into a 250ml Erlenmeyer flask, add 40ml of tetrahydrofuran, and stir at room temperature. Then drop in dilute hydrochloric acid (219mg, 6.01mmol) (dissolved in 4ml of water), heat to 60℃, dissolve until clear, drop into sodium hydroxide solution (576mg, 14.40mmol) (dissolved in 40ml of water), and cool to room temperature Warm stirring overnight. After drying, 0.76 g of solid was obtained, and the yield was 76.0%. The X-diffraction and DSC spectra were studied and compared, and the product was determined to be of crystalline form II.

Example 7

The type II crystalline product samples obtained in Example 1 were placed open and flat, and the stability of the samples under the conditions of light (4500 Lux), heating (40°C, 60°C), and high humidity (RH 75%, RH90%) were investigated. . The investigation sampling time is 5 days and 10 days, and the purity detected by HPLC is shown in Table 1.

The stability inspection results show that the type II crystalline sample of compound represented by formula (I) has good stability under light, high temperature and high humidity conditions under open conditions.

Example 8

The type II crystals of the compound represented by formula (I) prepared according to the method of Example 1 were ground, heated and tableted. The research results showed that the crystal form was stable. For detailed experimental data, see Table 2 below.

Since the graphs in this case are experimental data, they are not representative graphs of this case.

Therefore, there is no designated representative diagram in this case.

Claims (9)

A type II crystal of the compound represented by formula (I), characterized in that: Cu-Ka radiation is used to obtain an X-ray powder diffraction pattern represented by 2θ angles and interplanar spacing, and the crystal has the shape shown in Figure 1. X-ray powder diffraction pattern at about 5.84 (15.12), 6.16 (14.35), 10.75 (8.22), 12.56 (7.04), 12.96 (6.82), 16.25 (5.45), 17.24 (5.14), 18.97 (4.67), 20.22 (4.39), 21.74 (4.08), 22.99 (3.87), and 25.85 (3.44) have characteristic peaks,

A method for preparing type II crystals of the compound represented by formula (I) described in item 1 of the scope of patent application, the method includes the following steps: 1) adding any crystal form or amorphous form of the compound represented by formula (I) Add an acid to an appropriate amount of solvent, dissolve it until it is clear, add an alkali, and then crystallize. The solvent is selected from methanol, ethanol, isopropanol, acetone or methanol/water, ethanol/water, acetone/water, and isopropanol/water; 2) The crystals are filtered, washed, and dried. The method described in item 2 of the scope of patent application, wherein the acid is an inorganic acid. The method described in item 3 of the scope of patent application, wherein the acid is hydrochloric acid. According to the preparation method described in item 2 of the scope of patent application, the base is an inorganic base. The method according to item 5 of the scope of patent application, wherein the alkali is sodium hydroxide or potassium hydroxide. The method described in item 2 of the scope of patent application, wherein the solvent in step 1) is ethanol. A pharmaceutical composition containing the type II crystal of the compound represented by formula (I) as described in item 1 of the scope of patent application and a pharmaceutically acceptable carrier. A use of the type II crystal of the compound represented by formula (I) described in item 1 of the scope of patent application or the pharmaceutical composition described in item 8 of the scope of patent application, which is used in the preparation of drugs for the treatment of breast cancer.

Images