CN106432243B

CN106432243B - Crystal form of hedgehog signal pathway inhibitor and preparation method thereof

Info

Publication number: CN106432243B
Application number: CN201610595474.4A
Authority: CN
Inventors: 武乖利; 钟良; 张全良; 卢韵
Original assignee: Jiangsu Hengrui Medicine Co Ltd
Current assignee: Tianjin Hengrui Pharmaceutical Co ltd; Jiangsu Hengrui Medicine Co Ltd
Priority date: 2015-08-04
Filing date: 2016-07-26
Publication date: 2020-01-10
Anticipated expiration: 2036-07-26
Also published as: CN106432243A

Abstract

The invention relates to a crystal form of a hedgehog signal pathway inhibitor and a preparation method thereof. In particular, the invention relates to N- (2-ethyl-5- (6- (2- (trifluoromethyl) pyridin-3-yl) imidazo [1, 2-b)]Pyridazin-2-yl) phenyl) -1-methylcyclopropanecarboxamide (compound of formula (I)) form II crystals and a process for preparing the same. The II-type crystal of the compound of the formula (I) obtained by the invention has good crystal form stability and chemical stability, and the used crystallization solvent has low toxicity and low residue, and can be better used for clinical treatment.

Description

Crystal form of hedgehog signal pathway inhibitor and preparation method thereof

Technical Field

The present invention relates to crystalline forms of N- (2-ethyl-5- (6- (2- (trifluoromethyl) pyridin-3-yl) imidazo [1,2-b ] pyridazin-2-yl) phenyl) -1-methylcyclopropanecarboxamide and methods for its preparation.

Background

Malignant tumors have become one of the major diseases that are common and seriously threaten people's life and quality of life. With the continuous and intensive research on tumor genetics and molecular biology, a plurality of key signal pathways related to tumors in cells are discovered. In 1980, the Hedgehog (Hh) gene was first found in Drosophila. The Hedgehog signal path is found in the process of researching the mutation of the drosophila, the Hedgehog signal path is also called a Hedgehog signal path, the Hedgehog gene is a segmented polar gene, and the Hedgehog signal path is obtained because the mutated drosophila embryo is in a multi-hair-ball shape and exactly like a frightened Hedgehog; three hedgehog homologous genes exist in humans, including sonic hedgehog (SHH), indian hedgehog (IHH) and desert hedgehog (DHH). They play an important role in embryonic development, controlling the formation of many tissues and organs.

The Hedgehog signaling pathway is an important regulatory pathway for cellular physiological processes, is strictly regulated under normal conditions, and plays an important role in cell proliferation, cell differentiation, and embryo formation. In recent years, many diseases are gradually found to be related to the signal pathway, and when the Hedgehog signal pathway is abnormally activated, the Hedgehog signal pathway causes the occurrence and development of various tumors, including small cell lung cancer, esophagus cancer, stomach cancer, pancreatic cancer, colon cancer, prostate cancer and the like, which account for about 25% of human tumors. The research has found that some regulators and inhibitors of the Hedgehog signaling pathway, such as Hh interacting protein l (Hip1), growth-blocking specific gene 1(GAS-1), vesicle transporter RAB23, Bone Morphogenetic Protein (BMP), sterol compounds, etc., can inhibit proliferation, differentiation and induce apoptosis of cells in response to the Hedgehog signaling pathway, and the research results provide important clues for treating tumors. The Cyelopamine is a plant steroid alkaloid, is used as an antagonist of a Hedgehog signal path, and is a promising cancer treatment drug. Vismodegib is approved for marketing for the treatment of basal cell carcinoma, demonstrating the success of hedgehog signaling pathway targeting drug strategies. Therefore, the development of small-molecule hedgehog signaling pathway inhibitors has great development prospect for treating cancers in clinical work.

WO2012088411 discloses a novel hedgehog signaling pathway inhibitor having the structure shown in formula (I) as follows:

the compound shown in the formula (I) is used as a brand-new micromolecule hedgehog signal channel inhibitor, and the inhibition effect on a target in vitro and the drug effect in a mouse body are more than two times stronger than the Vismodegib; and animal pharmacokinetics results show that compared with a positive medicament Vismodegib, the in vivo accumulation condition of the compound shown in the formula (I) is greatly improved, and the compound has a great development prospect.

WO2012088411 does not extensively study the crystalline form of the compound. It is well known to those skilled in the art that the crystal structure of the pharmaceutically active ingredient often affects the chemical stability of the drug, and the difference of crystallization conditions and storage conditions may cause the crystal structure of the compound to change, sometimes accompanied by the generation of other forms of crystal forms. Therefore, the research on the drug crystal form, particularly the research on the drug stable crystal form, has very important significance on the subsequent development of the drug.

Disclosure of Invention

The invention aims to provide a crystal form of a compound shown as a formula (I) and a preparation method thereof.

A series of crystal products are obtained by the compound shown in the formula (I) under different crystallization conditions, and X-ray diffraction and DSC detection are carried out on the obtained crystal products, so that the compound shown in the formula (I) can obtain a stable crystal form under the specific crystallization conditions of the invention, and is named as type II crystal.

In one aspect, the invention provides form II crystals of a compound of formula (I). Characterized in that, using Cu-Ka radiation, an X-ray powder diffraction pattern expressed in terms of 2 theta angles and interplanar spacings is obtained, said crystal having an X-ray powder diffraction pattern as shown in FIG. 1, wherein peaks are characteristic at about 4.65(19.00), 6.39(13.81), 7.97(11.08), 9.57(9.23), 11.11(7.96), 12.75(6.94), 13.64(6.49), 15.72(5.63), 17.36(5.10), 18.41(4.82), 19.18(4.62), 21.38(4.15), 22.75(3.91), 23.15(3.84), 24.77(3.59), 27.21(3.27), 28.50(3.13), 29.48(3.03), 30.47(2.93), 31.62(2.83), 32.96 (2.72); and a DSC pattern showing melting endothermic peaks at around 116 ℃ and 192 ℃.

The invention discloses a method for preparing II-type crystals of a compound shown in a formula (I), which specifically comprises the following steps:

(1) heating and dissolving N- (2-ethyl-5- (6- (2- (trifluoromethyl) pyridine-3-yl) imidazo [1,2-b ] pyridazine-2-yl) phenyl) -1-methylcyclopropanecarboxamide solid with any crystal form or amorphous form in a proper amount of organic solvent, cooling and crystallizing;

(2) the crystals were filtered, washed and dried.

In a preferred embodiment of the present invention, in step (1), the organic solvent is selected from alcohols having 3 or less carbon atoms; more preferably, the organic solvent is methanol or ethanol.

The method of recrystallization is not particularly limited, and the recrystallization can be carried out by a usual recrystallization operation method. For example, the compound represented by the formula (I) may be dissolved in an organic solvent under heating, cooled, crystallized, and then filtered and dried to obtain the desired crystals. In particular, the filtered crystals are usually dried under reduced pressure at about 30 to 100 ℃ and preferably 40 to 60 ℃ under vacuum to remove the recrystallization solvent.

The crystal form of the obtained compound crystal shown as the formula (I) is researched by differential scanning thermal analysis (DSC) and X-diffraction pattern measurement, and the solvent residue of the obtained crystal is detected.

The compound crystal of formula (I) prepared by the method of the invention contains no or only a low content of residual solvent, and meets the limit requirement of residual solvent of related medical products specified by the national pharmacopoeia, so that the crystal of the invention can be better used as a medical active ingredient.

Researches show that the II-type crystal of the compound shown in the formula (I) prepared by the invention has good stability under the conditions of illumination, high temperature and high humidity, has stable crystal form under the conditions of grinding, pressure, heating and the like, can meet the medicinal requirements of production, transportation and storage, has stable, repeatable and controllable production process, and can be suitable for industrial production.

The form I crystals of the present invention may be conveniently used for the preparation of a pharmaceutical composition comprising the crystals of the compound represented by formula (I) and at least one pharmaceutically acceptable carrier, excipient or diluent.

Drawings

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

FIG. 2 is a DSC of form II crystals of the compound represented by the formula (I).

FIG. 3 is an X-ray powder diffraction pattern of a form III crystal of the compound represented by the formula (I).

FIG. 4 is a DSC of form III crystals of the compound represented by the formula (I).

Detailed Description

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

Test instrument for experiments

1. DSC spectrum

The instrument model is as follows: mettler Toledo DSC 1Staree System

And (3) purging gas: nitrogen gas

The heating rate is as follows: 10.0 ℃/min

Temperature range: 40-250 deg.C

2. X-ray diffraction spectrum

The instrument model is as follows: bruker D8Focus X-ray powder diffractometer

Ray: monochromatic Cu-Kalpha ray

The scanning mode is as follows: θ/2 θ, scan range: 2-40 °

Voltage: 40KV current: 40mA

Example 1

Adding (1.0g, 2.15mmol) of the compound shown in the formula (I) (prepared by the method disclosed in WO 2012088411) into a 25ml single-neck bottle, adding 10ml of ethanol, heating, refluxing, dissolving, stirring, cooling, crystallizing to room temperature, performing suction filtration, and drying to obtain 668mg of solid, wherein the yield is 66.8%. The X-ray diffraction pattern of the crystalline sample is shown in FIG. 1. The crystals had characteristic peaks at about 4.65(19.00), 6.39(13.81), 7.97(11.08), 9.57(9.23), 11.11(7.96), 12.75(6.94), 13.64(6.49), 15.72(5.63), 17.36(5.10), 18.41(4.82), 19.18(4.62), 21.38(4.15), 22.75(3.91), 23.15(3.84), 24.77(3.59), 27.21(3.27), 28.50(3.13), 29.48(3.03), 30.47(2.93), 31.62(2.83) and 32.96 (2.72). The DSC spectrum is shown in figure 2, and has sharp melting endothermic peaks at 116 ℃ and 192 ℃, and the crystal form is defined as the II crystal form.

Example 2

Adding (1.0g, 2.15mmol) of the compound shown in the formula (I) (prepared by the method disclosed in WO 2012088411) into a 25ml single-neck bottle, adding 8ml of methanol, heating, refluxing, dissolving, stirring, cooling, crystallizing to room temperature, performing suction filtration, and drying to obtain 606mg of solid, wherein the yield is 60.6%. The X-ray diffraction and DSC spectrogram of the crystallization sample are researched and compared to determine that the product is II crystal form.

Example 3

The entire procedure of example 633 of WO2012088411 was repeated, and 2-ethyl-5- (6- (2- (trifluoromethyl) pyridin-3-yl) imidazo [1,2-b ] pyridazin-2-yl) aniline (500mg, 1.30mol), acetonitrile (5ml) and pyridine (0.315ml) were added to the flask. The mixture was stirred and 1-methylcyclopropylcarboxylic acid chloride (169mg, 1.43mmol) was added, and the mixture was stirred at room temperature for 4 hours. Upon completion of the reaction, the mixture was concentrated on silica gel and purified by silica gel chromatography (0-80% ethyl acetate in dichloromethane) to give crystals of compound I (400mg, 66%). Through research and comparison of X-ray diffraction and DSC spectrogram of the crystallization sample, the crystallization sample is defined as a III crystal form, but the III crystal form can be a mixed crystal form, the DSC spectrogram shows that the III crystal form has two melting endothermic peaks at similar positions, and the mixed crystal form has the risk of possible crystal transformation, so that the absorption of the medicine is changed, and the clinical use is not recommended.

Example 4

The type II crystalline product obtained in example 1 was left open and laid flat, and the stability of the sample was examined under light (4500Lux), heat (40 ℃, 60 ℃), high humidity (RH 75%, RH 90%). The sampling times were considered to be 5 days and 10 days, and the purity by HPLC was found to be shown in Table 1.

Stability of type II Crystal samples of the Compound of formula (I) shown in Table 1

The stability examination result shows that the stability of the compound II type crystal sample shown in the formula (I) is not greatly influenced by illumination, high temperature and high humidity under the open placement condition.

Example 5

The compound II crystal of formula (I) prepared in example 1 was ground, heated and tabletted, and the results of the study showed that the crystal form was stable, and the detailed experimental data are shown in table 2 below.

Table 2 study on the specific stability of the II crystal form of the compound of formula (I)

Claims

1. Form II crystals of the compound represented by the formula (I) characterized by obtaining an X-ray powder diffraction pattern expressed in terms of 2-theta angles and interplanar spacings using Cu-Ka radiation, wherein there are characteristic peaks at about 4.65, 6.39, 7.97, 9.57, 11.11, 12.75, 13.64, 15.72, 17.36, 18.41, 19.18, 21.38, 22.75, 23.15, 24.77, 27.21, 28.50, 29.48, 30.47, 31.62, 32.96,

2. the form II crystal according to claim 1, characterized in that an X-ray powder diffraction pattern expressed in terms of 2 θ angle and interplanar spacing is obtained using Cu-Ka radiation as shown in FIG. 1.

3. A process for preparing form II crystals of a compound of formula (I) as defined in claim 1 or 2, comprising the steps of:

1) heating and dissolving N- (2-ethyl-5- (6- (2- (trifluoromethyl) pyridine-3-yl) imidazo [1,2-b ] pyridazine-2-yl) phenyl) -1-methylcyclopropanecarboxamide solid with any crystal form or amorphous form in a proper amount of organic solvent, cooling and crystallizing, wherein the organic solvent is methanol or ethanol;

2) the crystals were filtered, washed and dried.

4. A pharmaceutical composition comprising form II crystals of a compound of formula (I) according to claim 1 or 2 and at least one pharmaceutically acceptable carrier, excipient or diluent.

5. Use of the form II crystal of the compound of formula (I) according to claim 1 or 2 or the pharmaceutical composition of claim 4 in the preparation of a medicament for treating a disease associated with a small molecule hedgehog signaling pathway inhibitor.

6. Use according to claim 5, characterized in that the disease is selected from the group consisting of malignant tumors.

7. Use according to claim 5, characterized in that the disease is selected from basal cell carcinoma, gastric cancer, pancreatic cancer, lung cancer.