WO2023093859A1 - Salt of axl kinase inhibitor, preparation method therefor and use thereof - Google Patents

Abstract

The present invention provides a salt of a compound of formula (I) and a crystal form thereof, a preparation method therefor and use thereof. The salt is selected from the group consisting of mesylate, benzene sulfonate, oxalate, fumarate, citrate, equine urate, hydrochloride, hydrobromate, sulfate, or phosphate.

Description

Salts of AXL kinase inhibitors, their preparation methods and uses technical field

The invention belongs to the technical field of medicine, and the compound is an AXL kinase inhibitor, and specifically relates to a salt of the AXL inhibitor, a preparation method and a medical application thereof.

Background technique

Receptor tyrosine kinases (RTKs) are multidomain transmembrane proteins that serve as sensors of extracellular ligands. Ligand-receptor binding induces receptor dimerization and activation of its intracellular kinase domain, which in turn leads to the recruitment, phosphorylation and activation of multiple downstream signaling cascades (Robinson, D.R. et al., Oncogene, 19:5548-5557, 2000). To date, 58 RTKs have been identified in the human genome that regulate a variety of cellular processes, including cell survival, growth, differentiation, proliferation, adhesion, and motility (Segaliny, A.I. et al., J. Bone Oncol, 4:1 -12, 2015).

AXL (also known as UFO, ARK, and Tyro7) belongs to the TAM family of receptor tyrosine kinases, which also includes Mer and Tyro3. Among them, AXL and Tyro3 have the most similar gene structure, while AXL and Mer have the most similar amino acid sequence of tyrosine kinase domain. Like other receptor tyrosine kinases (RTKs), the structure of the TAM family consists of an extracellular domain, a transmembrane domain, and a conserved intracellular kinase domain. The extracellular domain of AXL has a unique structure that juxtaposes immunoglobulin and type III fibronectin repeat units and is reminiscent of a neutrophil adhesion molecule. TAM family members have a common ligand—growth arrest specific protein 6 (Gas6), which can bind to all TAM receptor tyrosine kinases. After AXL binds to Gas6, it will lead to receptor dimerization and AXL autophosphorylation, thereby activating multiple downstream signal transduction pathways and participating in multiple processes of tumorigenesis (Linger, R.M et al., Ther.Targets, 14(10 ), 1073-1090, 2010; Rescigno, J. et al., Oncogene, 6(10), 1909-1913, 1991).

AXL is widely expressed in normal tissues of the human body, such as monocytes, macrophages, platelets, endothelial cells, cerebellum, heart, skeletal muscle, liver, and kidney, among which the expression is highest in cardiac muscle and skeletal muscle, and bone marrow CD34+ cells and stromal cells also have a higher expression High expression, very low expression in normal lymphoid tissue (Wu YM, Robinson DR, Kung HJ, Cancer Res, 64(20), 7311-7320, 2004; hung BI et al., DNA Cell Biol, 22(8), 533-540 , 2003). In the study of many cancer cells, it was found that AXL gene was overexpressed or ectopically expressed in hematopoietic cells, mesenchymal cells and endothelial cells. In various leukemias and most solid tumors, the overexpression of AXL kinase is particularly prominent. Inhibition of AXL receptor tyrosine kinase can reduce the pro-survival signals of tumor cells, block the invasion ability of tumors, and increase the sensitivity of targeted drug therapy and chemotherapy. Therefore, finding effective AXL inhibitors is an important direction for the development of tumor-targeted drugs.

Contents of the invention

In one aspect, the present invention provides a pharmaceutically acceptable salt of the compound of formula I, wherein the salt is selected from organic acid salts or inorganic acid salts, wherein the organic acid salts are selected from methanesulfonate, benzenesulfonate, oxalate One of salt, fumarate, citrate and hippurate, the inorganic acid salt is selected from one of hydrochloride, hydrobromide, sulfate or phosphate, the compound of formula I The structure is as follows:

In some embodiments, the organic acid salt is mesylate.

In some embodiments, the mesylate salt is a hydrate of the mesylate salt.

In some embodiments, the mesylate salt is the dihydrate of the mesylate salt.

In some embodiments, the molar ratio of the compound of formula I to the organic acid in the organic acid salt is 1:1.

In some embodiments, the molar ratio of the compound of formula I to the inorganic acid in the inorganic acid salt is 1:1 or 1:2.

In some embodiments, the salt of an inorganic acid is a hydrochloride.

In some embodiments, the molar ratio of the compound of formula I to hydrogen chloride in the hydrochloride salt is 1:1 or 1:2.

In some embodiments, the molar ratio of the compound of formula I to hydrogen chloride in the hydrochloride salt is 1:2.

In some embodiments, the molar ratio of the compound of formula I to sulfuric acid in the sulfate salt is 1:1.

In some embodiments, the molar ratio of the compound of formula I to hydrobromic acid in the hydrobromide salt is 1:1.

In some embodiments, the molar ratio of the compound of formula I to phosphoric acid in the phosphate salt is 1:1.

It can be understood that the salt in the present invention is obtained by a salt-forming reaction between the compound of formula I and the corresponding acid. In the reaction, the compound of formula I is converted into a cation and combined with the acid radical of the corresponding acid to form the salt. Therefore, the molar ratio of the compound of formula I to the acid in the present invention can be understood as the molar ratio of the cation of the compound of formula I to the acid group of the corresponding acid in the salt.

In some typical embodiments, the present invention provides the mesylate salt of the compound of formula I, wherein the molar ratio of the compound of formula I to methanesulfonic acid is 1:1, or the molar ratio of the cation of the compound of formula I to the acid group of methanesulfonic acid The ratio is 1:1.

In another aspect, the present invention provides a crystal form of a pharmaceutically acceptable salt of the compound of formula I, wherein the salt is selected from organic acid salts or inorganic acid salts, wherein the organic acid salts are selected from methanesulfonate, benzenesulfonate , oxalate, fumarate, citrate and hippurate, the inorganic acid salt is selected from hydrochloride, hydrobromide or phosphate.

In some embodiments, the present invention provides a crystalline form of the mesylate salt of the compound of Formula I.

In some embodiments, the 2θ of the X-ray powder diffraction pattern of the mesylate salt crystal is detailed in Table 1 below:

Table 1 The mesylate salt crystal form X-ray powder diffraction pattern data of formula I compound

2θ(°)	height (counts)	I/I 0 (%)
5.300	54	17.1
7.793	46	14.5
9.174	102	32.0
10.479	60	18.9
13.271	83	26.2
15.415	318	100.0
16.067	90	28.4
17.164	89	28.1
17.693	80	25.0
18.201	41	12.8
18.959	83	26.1
20.332	212	66.7
20.853	38	11.8
21.883	99	31.0
22.519	26	8.2
23.101	117	36.9
23.484	51	16.0
24.949	34	10.6
25.792	37	11.5
26.331	28	8.7
29.544	82	25.9
33.006	26	8.2
39.463	71	22.3

.

In some embodiments, the present invention provides a crystalline form of the mesylate salt of the compound of formula I, the X-ray powder diffraction pattern of which is shown in FIG. 1 .

In some embodiments, the present invention provides a crystalline form of the monohydrochloride salt of a compound of formula I.

In some embodiments, the 2θ of the X-ray powder diffraction pattern of the monohydrochloride crystal is detailed in Table 2 below:

Table 2 The monohydrochloride crystal form X-ray powder diffraction pattern data of the compound of formula I

In some embodiments, the present invention provides a crystalline form of the monohydrochloride salt of the compound of formula I, the X-ray powder diffraction pattern of which is shown in FIG. 4 . In some embodiments, the present invention provides a crystalline form of the dihydrochloride salt of the compound of formula I.

In some embodiments, the dihydrochloride salt is crystalline, and the 2θ of its X-ray powder diffraction pattern is detailed in Table 3 below:

Table 3 The dihydrochloride salt crystal form X-ray powder diffraction pattern data of the compound of formula I

In some embodiments, the present invention provides a crystal form of the dihydrochloride salt of the compound of formula I, the X-ray powder diffraction pattern of which is shown in FIG. 5 .

In some embodiments, the present invention provides a crystalline form of the phosphate salt of a compound of formula I.

In some embodiments, the 2θ of the X-ray powder diffraction pattern of the phosphate crystal is detailed in Table 4 below:

Phosphate crystal form X-ray powder diffraction pattern data of the compound of formula I in table 4

2θ(°)	height (counts)	I/I 0 (%)
4.921	34	22.3
6.595	7	4.8
9.767	153	100.0
10.994	twenty three	15.4
11.732	twenty four	15.6
13.388	45	29.8
14.569	49	32.4
15.322	20	12.9
16.030	13	8.7
18.439	55	36.0
19.043	66	43.3
20.484	17	11.3
21.343	twenty two	14.2
22.584	46	29.9
24.366	82	53.8
24.890	60	39.0
25.994	60	39.1
29.305	72	47.3
32.096	7	4.9
33.988	13	8.3
35.573	9	5.9
37.022	12	7.9
39.391	9	5.8

.

In some embodiments, the present invention provides a crystalline form of the phosphate salt of the compound of formula I, the X-ray powder diffraction pattern of which is shown in FIG. 6 .

In some embodiments, the present invention provides a crystalline form of a hippurate salt of a compound of Formula I.

In some embodiments, the 2θ of the X-ray powder diffraction pattern of the hippurate crystal is detailed in Table 5 below:

Table 5 The hippurate crystal form X-ray powder diffraction pattern data of the compound of formula I

In some embodiments, the present invention provides a crystalline form of hippurate of the compound of formula I, the X-ray powder diffraction pattern of which is shown in FIG. 7 .

In some embodiments, the present invention provides an amorphous form of the sulfate salt of the compound of formula I.

In some embodiments, the present invention provides a crystalline form of the hydrobromide salt of a compound of formula I.

In some embodiments, the 2θ of the hydrobromide crystal is shown in Table 6 below in detail in its X-ray powder diffraction pattern:

The hydrobromide salt crystal form X-ray powder diffraction pattern data of table 6 formula I compound

2θ(°)	height (counts)	I/I 0 (%)
5.671	15	6.6
8.457	56	24.2
11.318	145	62.5
12.029	69	29.9
12.920	205	88.2
13.219	110	47.4
13.753	181	78.2
14.791	42	17.9
16.779	143	61.5
18.057	112	48.3
21.010	141	61.0
21.484	121	52.1
22.056	62	26.6
22.614	98	42.2
23.270	232	100.0
24.129	181	77.9
24.907	94	40.4
25.636	148	63.7
27.155	100	43.0
28.296	57	24.7
29.464	32	13.7
33.019	32	13.6
34.699	67	28.8

.

In some embodiments, the present invention provides a crystal form of the hydrobromide salt of the compound of formula I, the X-ray powder diffraction pattern of which is shown in FIG. 9 .

In some embodiments, the present invention provides a crystalline form of the besylate salt of the compound of formula I.

In some embodiments, the benzenesulfonate salt is crystalline, and the 2θ of its X-ray powder diffraction pattern is detailed in Table 7 below:

Table 7 Besylate crystal form X-ray powder diffraction pattern data

2θ(°)	height (counts)	I/I 0 (%)
10.980	25	12.3
12.144	90	43.7
13.601	60	29.5
15.200	125	60.8
17.066	116	56.6
18.536	205	100.0
19.384	99	48.5
20.160	111	54.0
21.250	73	35.7
21.975	203	99.2
24.505	154	75.0
26.759	78	38.0
27.897	47	23.1
30.580	36	17.5
32.115	18	8.6

.

In some embodiments, the present invention provides a crystalline form of the besylate salt of the compound of formula I, the X-ray powder diffraction pattern of which is shown in FIG. 10 .

In some embodiments, the present invention provides a crystalline form of the oxalate salt of the compound of formula I.

In some embodiments, the oxalate salt crystals, the 2θ of its X-ray powder diffraction pattern are detailed in Table 8 below:

Table 8 X-ray powder diffraction pattern data of the oxalate salt crystal form of the compound of formula I

2θ(°)	height (counts)	I/I 0 (%)
5.912	94	14.6
8.834	174	26.9
10.270	114	17.7
11.760	78	12.1
14.710	646	100.0
17.656	485	75.0
18.229	72	11.2
19.318	54	8.4
19.720	60	9.4
21.564	194	30.0
25.441	103	16.0
26.590	121	18.7
27.016	100	15.5
32.375	7	1.0
34.957	29	4.5

.

In some embodiments, the present invention provides the crystalline form of the oxalate salt of the compound of formula I, the X-ray powder diffraction pattern of which is shown in FIG. 11 .

In some embodiments, the present invention provides a crystalline form of the fumarate salt of the compound of formula I.

In some embodiments, the 2θ of the X-ray powder diffraction pattern of the fumarate crystals is detailed in Table 9 below:

Table 9 Fumarate salt crystal form X-ray powder diffraction pattern data of the compound of formula I

In some embodiments, the present invention provides a crystalline form of the fumarate salt of the compound of formula I, the X-ray powder diffraction pattern of which is shown in FIG. 12 .

In some embodiments, the present invention provides a crystalline form of the citrate salt of the compound of formula I.

In some embodiments, the 2θ of the X-ray powder diffraction pattern of the citrate crystals is detailed in Table 10 below:

The citrate crystal form X-ray powder diffraction pattern data of the compound of formula I in table 10

2θ(°)	height (counts)	I/I 0 (%)
18.138	90	100.0
19.467	63	70.0
26.072	55	61.5
31.101	14	15.4

.

In some embodiments, the present invention provides a crystalline form of the citrate salt of the compound of formula I, the X-ray powder diffraction pattern of which is shown in FIG. 13 .

In another aspect, the present invention provides a crystal form A of the compound of formula I, whose X-ray powder diffraction pattern is 7.6°±0.2°, 10.2°±0.2°, 17.6°±0.2°, 20.3°±0.2° at 2θ There are diffraction peaks at 20.9°±0.2°.

Further, the crystal form A has an X-ray powder diffraction pattern at 2θ of 4.1°±0.2°, 7.6°±0.2°, 10.2°±0.2°, 12.6°±0.2°, 13.0°±0.2°, 17.6° There are diffraction peaks at ±0.2°, 19.7°±0.2°, 20.3°±0.2°, 20.9°±0.2° and 22.2°±0.2°.

Further, the crystal form A has an X-ray powder diffraction pattern at 2θ of 4.1°±0.2°, 5.6°±0.2°, 7.6°±0.2°, 10.2°±0.2°, 10.9°±0.2°, 12.6° ±0.2°, 13.0°±0.2°, 15.2°±0.2°, 17.6°±0.2°, 19.7°±0.2°, 20.3°±0.2°, 20.9°±0.2°, 22.2°±0.2°, 23.2°±0.2 °, 24.6°±0.2°, 27.0°±0.2°, 28.8°±0.2°, 37.0°±0.2° and 37.7°±0.2° have diffraction peaks.

In some embodiments, the 2θ of the X-ray powder diffraction pattern of the crystal form A is detailed in the table below:

Table 11 X-ray powder diffraction pattern data of crystal form A

In some embodiments, the X-ray powder diffraction of the crystal form A in 2θ angle has a pattern as shown in FIG. 14 .

In another aspect, the present invention provides a method for preparing a pharmaceutically acceptable salt of the compound of formula I or a crystal form of the pharmaceutically acceptable salt thereof, which comprises the step of forming a salt of the compound of formula I with a corresponding acid.

In some embodiments, the reaction solvent of the preparation method is selected from mixed solvents of alcohol solvents and alkane solvents, mixed solvents of ketone solvents and alkane solvents, mixed solvents of ester solvents and alkane solvents, nitrile solvents - A mixed solvent of an aqueous solvent and an alkane solvent, a mixed solvent of an alkylbenzene solvent and an alkane solvent, or a mixed solvent of a halogenated hydrocarbon solvent and an alkane solvent.

In some embodiments, the alcohol solvent is selected from methanol, ethanol or isopropanol; the ketone solvent is selected from acetone or butanone; preferably acetone; the ester solvent is selected from ethyl acetate or butyl acetate ; Preferred ethyl acetate; The nitriles-water solvent is selected from the mixed solution of nitriles-water, and the alkane solvent is selected from n-heptane.

In some embodiments, the crystal form of the pharmaceutically acceptable salt is a mesylate salt crystal form, and the mesylate salt crystal form is prepared by a method comprising the following steps:

S1: stirring the compound of formula I, methanesulfonic acid and toluene to obtain a seed crystal of the mesylate salt crystal form of the compound of formula I;

S2: stirring and reacting the compound of formula (I), methanesulfonic acid and ethyl acetate, adding the crystal form obtained in step S1 and continuing to stir;

S3: adding ethyl acetate, acetone and the crystal form of the compound of formula I according to claim 8, stirring and crystallizing to obtain the mesylate salt crystal form (or called mesylate salt crystal form II).

In another aspect, the present invention also provides a pharmaceutical composition comprising a pharmaceutically acceptable salt of the compound of formula I.

In some embodiments, the pharmaceutical composition further comprises one or more pharmaceutically acceptable carriers.

In some embodiments, the pharmaceutical composition is a solid pharmaceutical preparation suitable for oral administration, preferably a tablet or a capsule.

In another aspect, the present invention also provides a pharmaceutical composition comprising the crystal form of the pharmaceutically acceptable salt of the compound of formula I.

In some embodiments, the pharmaceutical composition further comprises one or more pharmaceutically acceptable carriers.

In some embodiments, the pharmaceutical composition is a solid pharmaceutical preparation suitable for oral administration, preferably a tablet or a capsule.

In another aspect, the present invention provides a crystal form composition, wherein the pharmaceutically acceptable salt of the compound of formula I above, and the crystal form of the pharmaceutically acceptable salt of the compound of formula I account for the weight of the crystal form composition above 50.

Further accounting for more than 80% of the weight of the crystal composition;

Further accounting for more than 90% of the weight of the crystal composition;

It further accounts for more than 95% of the weight of the crystal composition.

On the other hand, the present invention also provides a pharmaceutical composition comprising the above crystal composition; the pharmaceutical composition further comprises one or more pharmaceutically acceptable carriers.

On the other hand, the present invention also provides a pharmaceutically acceptable salt of the compound of formula I or its crystal form composition or its pharmaceutical composition for use as a medicine.

On the other hand, the present invention also provides a pharmaceutically acceptable salt crystal form of the compound of formula I or its crystal form composition or pharmaceutical composition thereof for use as a medicine.

In another aspect, the present invention also provides a method for preventing and/or treating AXL kinase-mediated diseases or disease states, which comprises administering the salt of the compound of formula I of the present invention, or its crystal form, to individuals in need Composition or its pharmaceutical composition.

On the other hand, the present invention also provides a method for preventing and/or treating AXL kinase-mediated diseases or disease states, which comprises administering the crystalline form of the salt of the compound of formula I of the present invention, Its crystal form composition or its pharmaceutical composition.

On the other hand, the present invention also provides the salt of the compound of formula I of the present invention, its crystal form composition or its pharmaceutical composition for preventing and/or treating AXL kinase-mediated diseases or disease states.

In another aspect, the present invention also provides the crystalline form of the salt of the compound of formula I of the present invention, its crystalline form composition or its pharmaceutical composition for preventing and/or treating AXL kinase-mediated diseases or disease states .

In some embodiments, the AXL kinase-mediated disease or condition is cancer.

In some exemplary embodiments, the cancer is a disease associated with hematological and solid tumors.

related definition

Unless otherwise specified, the following terms used in the specification and claims have the following meanings:

The pharmaceutically acceptable salts of the present invention also include their hydrated forms.

The term "pharmaceutically acceptable carrier" refers to those carriers that have no obvious stimulating effect on the body and will not impair the biological activity and performance of the active compound. Including but not limited to any diluents, disintegrants, binders, glidants, and wetting agents approved by the State Food and Drug Administration for human or animal use.

The term "fumaric acid" refers to fumaric acid, which has the structure:

The term "alcohol solvent" refers to a substance derived from one or more hydroxyl groups (OH) replacing one or more hydrogen atoms on a C1-C6 alkane, and the C1-C6 alkane refers to a substance containing 1-6 carbon atoms Specific examples of linear or branched alkanes and alcohol solvents include, but are not limited to: methanol, ethanol, isopropanol or n-propanol.

The term "alkane solvent" refers to straight chain or branched chain or cyclic alkanes containing 5-7 carbon atoms, specific examples include but not limited to n-hexane, cyclohexane, n-heptane.

The term "ester solvent" refers to a chain compound containing an ester group -COOR and a carbon number of 3-10, wherein R is a C1-C6 alkyl group, and the C1-C6 alkyl group refers to a chain compound containing 1-6 Specific examples of straight-chain or branched-chain alkanes with carbon atoms and ester solvents include but are not limited to methyl acetate, ethyl acetate, and propyl acetate.

The term "halogenated hydrocarbon solvent" refers to a substance derived from one or more halogen atoms replacing one or more hydrogen atoms on a C1-C6 alkane, and the C1-C6 alkane refers to a substance containing 1-6 carbon atoms For linear or branched alkanes, the halogen atoms refer to fluorine, chlorine, bromine, and iodine. Specific examples of halogenated hydrocarbon solvents include but are not limited to dichloromethane or chloroform.

The term "ketone solvent" refers to a chain or cyclic compound containing carbonyl -CO- and having 3-10 carbon atoms, specific examples include but not limited to acetone, methyl ethyl ketone or cyclohexanone.

The term "benzene-based solvent" means a solvent containing a phenyl group, and specific examples include toluene, xylene, cumene, or chlorobenzene.

The term "equivalent" refers to the equivalent amount of other raw materials required in accordance with the equivalent relationship of chemical reactions, taking the basic raw materials used in each step as 1 equivalent.

The "X-ray powder diffraction pattern" in the present invention is obtained by using Cu-Kα radiation measurement.

It should be noted that in X-ray powder diffraction (XRPD), the diffraction pattern obtained from a crystalline compound is often characteristic for a particular crystal, where the relative intensity of the bands (especially at low angles) may vary due to The effect of dominant orientation due to differences in crystallization conditions, particle size and other measurement conditions varies. Therefore, the relative intensities of the diffraction peaks are not characteristic of the targeted crystals. When judging whether it is identical to a known crystal, more attention should be paid to the relative positions of the peaks rather than their relative intensities. Furthermore, for any given crystal there may be slight errors in the position of the peaks, as is well known in the art of crystallography. For example, due to temperature changes, sample movement, or instrument calibration when analyzing samples, the position of the peak can move, and the measurement error of the 2θ value is sometimes about ±0.2°. Therefore, this error should be taken into account when determining each crystal structure. In the XRPD spectrum, the 2θ angle or the crystal plane distance d is usually used to represent the peak position, and there is a simple conversion relationship between the two: d=λ/2sinθ, where d represents the crystal plane distance, λ represents the wavelength of the incident X-ray, and θ is Diffraction angle. For the same crystal of the same compound, the peak positions of their XRPD spectra are similar on the whole, and the relative intensity error may be large. It should also be pointed out that in the identification of mixtures, due to factors such as content decline, some diffraction lines will be missing. At this time, it is not necessary to rely on all the bands observed in the high-purity sample, and even one band may affect the given Certain crystallization is characteristic.

Differential Scanning Calorimetry (DSC) measures the transition temperature when a crystal absorbs or releases heat due to a change in its crystal structure or melting of the crystal. For the same crystal form of the same compound, thermal transition temperatures and melting points are typically within about 5°C, usually within about 3°C, in successive analyses. When it is described that a compound has a given DSC peak or melting point, it is meant that the DSC peak or melting point ± 5°C. DSC provides an auxiliary method to distinguish different crystal forms. Different crystal forms can be identified by their different transition temperature characteristics. It should be pointed out that for the mixture, its DSC peak or melting point may fluctuate in a larger range. In addition, since the melting process of the substance is accompanied by decomposition, the melting temperature is related to the heating rate.

Thermogravimetric analysis (TGA) refers to a thermal analysis technique that measures the relationship between the mass of the sample to be tested and the temperature change at a programmed temperature. When the measured substance sublimes or vaporizes during the heating process, it decomposes into gas or loses crystal water, causing the quantity of the measured substance to change. At this time, the thermogravimetric curve is not a straight line but a decline. By analyzing the thermogravimetric curve, you can know at what temperature the measured substance changes, and according to the lost weight, you can calculate how much the substance is lost.

When referring to, for example, an XRPD pattern, DSC pattern or TGA pattern, the term "as shown" includes patterns that are not necessarily identical to those depicted herein, but which fall within the limits of experimental error when considered by those skilled in the art .

Unless otherwise specified, the abbreviation of the present invention has the following meanings:

M: mol/L

mM: mmol/L

nM: nmol/L

Boc: tert-butoxycarbonyl

¹ H NMR: Proton Magnetic Resonance Spectroscopy

MS(ESI+): mass spectrum

DMSO-d ₆ : deuterated dimethyl sulfoxide

CDCl ₃ : deuterated chloroform

DTT: Dithiothreitol

SEB: Supplemented Enzymatic Buffer (Supplemented Enzymatic Buffer)

IMDM (Iscove's Modified Dulbecco's Medium): Iscove (person's name) modified Dulbecco (person's name) medium.

Room temperature: 25°C.

Description of drawings

In order to more clearly illustrate the embodiments of the present invention and the technical solutions of the prior art, the following briefly introduces the accompanying drawings that need to be used in the embodiments and the prior art. Obviously, the accompanying drawings in the following description are only examples For some embodiments of the invention, those skilled in the art can also obtain other drawings according to these drawings.

Fig. 1 shows the X-ray powder diffraction (XRPD) spectrogram of formula I compound mesylate in embodiment 5;

Fig. 2 shows the differential scanning calorimetry (DSC) figure of formula I compound mesylate in embodiment 5;

Fig. 3 shows the thermogravimetric (TGA) figure of formula I compound mesylate in embodiment 5;

Fig. 4 shows the X-ray powder diffraction (XRPD) spectrogram of formula I compound monohydrochloride in embodiment 3;

Fig. 5 shows the X-ray powder diffraction (XRPD) spectrogram of formula I compound dihydrochloride in embodiment 3;

Fig. 6 shows the X-ray powder diffraction (XRPD) spectrogram of formula I compound phosphate in embodiment 3;

Fig. 7 shows the X-ray powder diffraction (XRPD) spectrogram of formula I compound hippurate in embodiment 3;

Fig. 8 shows the X-ray powder diffraction (XRPD) spectrogram of formula I compound sulfate in embodiment 3;

Fig. 9 shows the X-ray powder diffraction (XRPD) spectrogram of formula I compound hydrobromide in embodiment 3;

Fig. 10 shows the X-ray powder diffraction (XRPD) spectrogram of formula I compound besylate in embodiment 3;

Fig. 11 shows the X-ray powder diffraction (XRPD) spectrogram of formula I compound oxalate in embodiment 3;

Fig. 12 shows the X-ray powder diffraction (XRPD) spectrogram of formula I compound fumarate in embodiment 3;

Fig. 13 shows the X-ray powder diffraction (XRPD) spectrogram of formula I compound citrate in embodiment 3;

Figure 14 shows the X-ray powder diffraction (XRPD) spectrum of the crystal form A of the compound of formula I in Example 6.

Detailed ways

The present invention is described in more detail below by way of examples. However, these specific descriptions are only used to illustrate the technical solution of the present invention, and do not constitute any limitation to the present invention.

The test conditions of each instrument are as follows:

(1) X-ray powder diffractometer (X-ray Powder Diffraction, XRPD)

Instrument model: Bruker D2 Phaser 2 ^nd

(2) Thermogravimetric Analyzer (Thermogravimetric, TGA)

Instrument model: TA Instruments TGA25

Purge gas: Nitrogen

Heating rate: 10°C/min

Temperature rise range: room temperature - 300°C

Method: Put the sample in an aluminum pan, then place the aluminum pan in a platinum pan, expose it in a nitrogen atmosphere, and raise the temperature from room temperature to the set temperature at a rate of 10°C/min.

(3) Differential Scanning Calorimeter (DSC)

Instrument model: TA Instruments DSC25

Purge gas: Nitrogen

Heating rate: 10°C/min

Heating range: 20-300°C

Method: The sample was placed in an aluminum pan, and after capping, the temperature was raised from 20°C to the set temperature at a rate of 10°C/min in a nitrogen atmosphere.

(4) Dynamic moisture adsorption (DVS)

Instrument model: Surface Measurement System (SMS)-DVS Intrinsic

The specific instrument setting parameters are as follows:

Example 1 (S)-(2-((5-chloro-2-((7-(pyrrolidin-1-yl)-6,7,8,9-tetrahydro-5H-benzo[7] wheel Preparation of En-2-yl)amino)pyrimidin-4-yl)amino)-5-(methoxymethyl)phenyl)dimethylphosphine oxide

a) 2-iodo-4-(methoxymethyl)aniline

4-(Methoxymethyl)aniline (9g), iodine (16.65g) and sodium bicarbonate (16.53g) were added to a solution of dichloromethane (261mL)/water (135mL), and stirred at 22°C for 16h. The reaction was quenched with saturated sodium thiosulfate (10 ml) at room temperature. The resulting mixture was extracted with dichloromethane (3 x 100 mL), then the combined organic layers were washed with saturated aqueous sodium chloride (1 x 100 mL), and the organic layer was dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate=1/1 v/v) to obtain the title product (16 g). MS(ESI+):264.0(M+H).

b) (2-Amino-5-(methoxymethyl)phenyl)dimethylphosphine oxide

To N,N-dimethylformamide (224 mL) was added 2-iodo-4-(methoxymethyl)aniline (16 g, 60.82 mmol, 1.00 equiv), potassium phosphate (14.20 g) under nitrogen atmosphere , palladium acetate (0.68g) and 4,5-bisdiphenylphosphine-9,9-dimethylxanthene (1.76g) in a stirred solution was added dimethylphosphine oxide (5.22g), at 120 ° C The reaction was stirred for 2 hours. The mixture was cooled to room temperature. The resulting mixture was filtered and the filter cake was washed with N,N-dimethylformamide (3x5 mL). The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (dichloromethane/methanol=20/1 v/v) to obtain the title product (12.9 g). MS(ESI+):214.1(M+H).

c) (2-((2,5-dichloropyrimidin-4-yl)amino)-5-(methoxymethyl)phenyl)dimethylphosphine oxide

To N,N-dimethylformamide (22 mL) was added (2-amino-5-(methoxymethyl)phenyl)dimethylphosphine oxide (1.10 g), 2,4,5 - Trichloropyrimidine (1.23g) and N,N-diisopropylethylamine (2.00g) were stirred for 3h. The resulting mixture was diluted with dichloromethane (30 mL). The reaction was quenched by adding water (10ml) at 0°C. The resulting mixture was extracted with dichloromethane (3 x 50 mL). The combined organic layers were washed with saturated sodium chloride (1x50 mL) and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (dichloromethane/methanol=20/1 v/v) to obtain the title product (1.28 g).

MS(ESI+):360.0(M+H).

d) (S)-(2-((5-chloro-2-((7-(pyrrolidin-1-yl)-6,7,8,9-tetrahydro-5H-benzo[7]annulene -2-yl)amino)pyrimidin-4-yl)amino)-5-(methoxymethyl)phenyl)dimethylphosphine oxide

To isopropanol (2 mL) was added (2-((2,5-dichloropyrimidin-4-yl)amino)-5-(methoxymethyl)phenyl)dimethylphosphine oxide (50.00 mg) and (S)-7-(pyrrolidin-1-yl)-6,7,8,9-tetrahydro-5H-benzo[7]cycloalken-2-amine (31.98mg), then added 1 of hydrogen chloride , 4-dioxane solution (10 drops, 4M), irradiated with microwave at 130°C for 3.5 hours. The mixture was then cooled to room temperature and concentrated under reduced pressure. The crude product was purified by reverse-phase high-performance liquid chromatography (column is YMC Actus Triart C18, 30*150mm, particle diameter 5 μm, mobile phase A: water (10mmol/L ammonium bicarbonate), mobile phase B: acetonitrile, flow rate: 60mL/ min, gradient: 20%B to 50%B, 8min, wavelength: 220nm, retention time: 6.83min, column temperature: 25°C), the title product (20.2mg) was obtained.

¹ H NMR (400MHz, DMSO-d ₆ , ppm): δ11.07(s, 1H), 9.26(s, 1H), 8.52(d, J=4.6Hz, 1H), 8.17(s, 1H), 7.53 (dd, J=14.0,2.0Hz,1H),7.44(q,J=3.1Hz,2H),7.26(dd,J=8.1,2.3Hz,1H),6.97(d,J=8.1Hz,1H) ,4.42(s,2H),3.31(s,3H),3.01–2.75(m,2H),2.55(s,5H),2.50(s,2H),1.84(s,2H),1.81(s,3H ),1.77(s,3H),1.70(q,J=3.6,3.2Hz,4H),1.54(s,2H).MS(ESI+):554.2(M+H).

Example 2 Activity Determination

Related compounds prepared in Example 1 carry out related enzyme activities, cells, and related activities in vivo

The specific structure of the positive drug 1 (BGB324) used in the activity test is as follows:

The specific structure of positive drug 2 (TP0903) is as follows:

All the above compounds were purchased from Shanghai Shenghong Biotechnology Co., Ltd.

(1) AXL kinase inhibitory activity

1. Experimental process

a) AXL enzyme (Carna, 08-107) configuration and addition: use 1× enzyme buffer (200 μL of Enzymatic buffer kinase 5X, 10 μL of 500 mM MgCl ₂ , 10 μL of 100 mM DTT, 6.26 μL of 2500 nM SEB, Add 773.75 μL of H ₂ O to make 1ml of 1×enzyme buffer) Dilute 33.33ng/uL of AXL enzyme to 0.027ng/μL (1.67×, final conc.=0.016ng/μL), use BioTek (MultiFlo FX) Automatic dispenser, add 6 μL of 1.67-fold final concentration of enzyme solution to compound wells and positive control wells; add 6 μL of 1×Enzymatic buffer to negative control wells.

b) Compound preparation and addition: use DMSO to dilute the compounds and positive drugs prepared in the examples from 10 mM to 100 μM, and titrate with a compound titrator (Tecan, D300e), and the titrator will automatically spray the required concentration into each well, the first step A concentration of 1 μM, 1/2 log gradient dilution, a total of 8 concentrations. Centrifuge at 2500rpm for 30s and incubate at room temperature for 15min.

c) ATP, substrate preparation and addition: ATP (Sigma, A7699) was diluted with 1× enzyme buffer, from 10 mM to 75 μM (5×), and the final concentration was 15 μM; substrate TK Substrate 3-biotin (Cisbio, 61TK0BLC) was diluted with 1× enzyme buffer solution from 500 μM to 5 μM (5×), and the final concentration was 1 μM; ATP was mixed with the substrate in equal volume, and 4 μL was added to each well using a BioTek automatic liquid dispenser; centrifuged at 2500 rpm for 30 s, at 25 ° C React for 45 minutes.

d) Detection reagent preparation and addition: Streptavidin-XL665 (Cisbio, 610SAXLG) was diluted from 16.67μM to 250nM (4×) with HTRF KinEASE detection buffer (cisbio), and the final concentration was 62.5nM; TK Antibody-Cryptate (Cisbio) was diluted with HTRF KinEASE detection buffer (cisbio) was diluted from 100× to 5×, and the final concentration was 1×; XL665 was mixed with Antibody in equal volume, 10 μL was added to each well using a BioTek automatic dispenser, centrifuged at 2500 rpm for 30 seconds, and reacted at 25°C for 1 hour. After the reaction, the multifunctional plate reader HTRF was used for detection.

2. Data analysis

Use GraphPad Prism 5 software log(inhibitor) vs.response-Variable slope to fit the dose-effect curve, and obtain the IC ₅₀ value of the compound on AXL kinase inhibition.

The formula for calculating the inhibition rate is as follows:

3. The experimental results are detailed in the table below

Table 12 Compound AXL inhibitory activity IC ₅₀ data

(2) Detection of compound's inhibition of cell proliferation

1. Experimental process

MV-4-11 (human myelomonocytic leukemia cell line, medium: IMDM+10% fetal bovine serum) was purchased from Nanjing Kebai Biotechnology Co., Ltd., and placed in an incubator at 37°C and 5% CO ₂ nourish. Cells in the logarithmic growth phase were plated in 96-well plates at cell densities of 8000/well, 6000/well, 5000/well, 4000/well and 3000/well, and a blank control group was set at the same time.

Dissolve the test compound and the positive drug in dimethyl sulfoxide to prepare a 10 mM stock solution, and store it in a -80°C refrigerator for long-term storage. 24 hours after cell plating, dilute the 10 mM compound stock solution with dimethyl sulfoxide to obtain a 200-fold working solution (maximum concentration 200 or 2000 μM, 3-fold gradient, 10 concentrations in total), and add 3 μL of each concentration to 197 μL In the complete culture medium, dilute to obtain a 3-fold working solution, then take 50 μL and add it to 100 μL cell culture medium (the final concentration of dimethyl sulfoxide is 0.5%, v/v), and set two replicates for each concentration. hole. After 72 hours of drug treatment, add 50 μl of

(purchased from Promega), the fluorescent signal was measured on Envision (PerkinElmer) according to the operating procedures of the instructions, and the dose-effect curve was fitted using GraphPad Prism 5 software log (inhibitor) vs. response-Variable slope to obtain the IC of the compound on cell proliferation inhibition ₅₀ value. Inhibition rate calculation formula:

in:

Signal value of the test substance: the mean value of the fluorescent signal of the cell + medium + compound group;

Signal value of the blank group: the average value of the fluorescence signal of the culture medium group (containing 0.5% DMSO);

Signal value of negative control group: mean value of fluorescence signal of cell+medium group (containing 0.5% DMSO).

2. Experimental results

The IC ₅₀ (MV4-11, nM) of the antiproliferative activity of the compound of Example 1 on MV4-11 cells was 6.97.

(3) MV4-11 in vivo efficacy of the compound

The inhibitory effect of the test compound and the positive drug on the growth of human acute monocytic leukemia cell MV-4-11 xenografted tumor model in nude mice in vivo.

1. Construction of mouse model

MV-4-11 cells in the logarithmic growth phase were collected, counted and resuspended, and the cell concentration was adjusted to 7.0×10 ⁷ cells/mL; injected subcutaneously into the right axilla of nude mice, each animal was inoculated with 200 L (14 ×10 ⁶ cells/monkey), the MV-4-11 xenograft tumor model was established. When the tumor volume reaches 100-300 mm ³ , tumor-bearing mice with good health and similar tumor volume are selected.

2. Compound Configuration

Vortex the compound and the positive drug with an appropriate solvent, then ultrasonically dissolve the compound completely, then slowly add an appropriate volume of citrate buffer, vortex, and mix the liquid evenly to obtain the concentration of 0.1, 0.5, 1 mg mL ^-1 Dosing preparations.

Solvent control group: PEG400&citric acid buffer (20:80, v:v).

3. Grouping and administration of animals

The modeled mice were randomly grouped (n=6), and related compounds and positive drugs were administered on the day of grouping, and the experiment was ended after 21 days or when the tumor volume of the solvent control group reached 2000 ^mm (whichever was reached first), and the administration volume Both are 10 mL·kg ^-1 . Both the compound and the positive drug were administered by intragastric administration, once a day. After the experiment started, the tumor diameter and animal body weight were measured twice a week, and the tumor volume was calculated.

4. Data Analysis

The formula for calculating tumor volume (TV) is: tumor volume (mm ³ )=l×w ² /2,

Among them, l represents the long diameter of the tumor (mm); w represents the short diameter of the tumor (mm).

The formula for calculating the relative tumor volume (RTV) is: RTV=TV _t /TV _initial

Among them, TV _initial is the tumor volume measured during group administration; TV _t is the tumor volume at each measurement during administration.

The calculation formula of tumor growth inhibition rate TGI(%) is: TGI＝100%×[1－(TV _t(T) －TV _initial(T) )/(TV _t(C) －TV _initial(C) )]

Among them, TV _{t (T)} represents the tumor volume measured each time in the treatment group; TV _{initial (T)} represents the tumor volume of the treatment group when administered in groups; TV _{t (C)} represents the tumor volume measured each time in the solvent control group; TV _{initial (C)} represents the tumor volume of the solvent control group at the time of group administration.

The formula for calculating the relative tumor proliferation rate (%T/C) is: %T/C=100%×(RTV _T /RTV _C )

Among them, RTV _T represents the RTV of the treatment group; RTV _C represents the RTV of the solvent control group.

The experimental data were calculated and related statistical processing with Microsoft Office Excel 2007 software.

5. The experimental results are shown in the table below:

In vivo efficacy of the compound of table 13

Remarks: The experimental data in the table is the relevant data obtained when the experiment ends (the end of the experiment is defined as: after 21 days or when the tumor volume of the solvent control group reaches 2000 mm ³ and the experiment ends (whichever is reached earlier)).

(4) Pharmacokinetic study of compounds in ICR mice

1. Prescription configuration for gavage of compounds

Each compound was prepared as a 10 mg/mL stock solution in DMSO.

Mixed solvent preparation: Tween 80:PEG400:Water＝1:9:90(v/v/v)

Accurately draw 450 μl of the compound DMSO stock solution with a concentration of 10 mg/mL into a glass bottle, add an appropriate volume of DMSO and a mixed solvent, the ratio of the solvent in the final preparation is DMSO:mixed solvent (v/v)=10:90, vortex (or ultrasound), and disperse evenly to obtain 4.5mL administration test solutions with a concentration of 1mg/mL respectively.

2. Test plan

Male ICR mice aged 6-10 weeks (source of mice: Weitong Lihua Experimental Animal Technology Co., Ltd.), 6 in each group, were fasted overnight and fed 4 hours after administration. On the day of the experiment, the mice were given 10 mg kg- ¹ compound test solution by intragastric administration. At 0, 5min, 15min, 30min, 1h, 2h, 4h, 8h, and 24h after administration, about 100 μL of blood was collected from the orbit of the mice and placed in an EDTA-K ₂ anticoagulant tube. Whole blood samples were centrifuged at 1500-1600 g for 10 min, and the separated plasma was stored in a -40-20°C refrigerator for biological sample analysis. LC-MS/MS method was used to determine the plasma concentration.

3. Data analysis and results

The pharmacokinetic parameters were calculated using the non-compartmental model in Pharsight Phoenix 7.0, and the specific results are shown in the table below.

The mouse pharmacokinetic result of table 14 compound

compound	Cmax(ng/mL)	Tmax(h)	AUC0-24(ng.h/mL)	T 1/2 (h)
Example 1	324	2.17	1300	1.35
Positive drug 2 (TP-0903)	26.8	0.25	52.2	1.20

Example 3 (S)-(2-((5-chloro-2-((7-(pyrrolidin-1-yl)-6,7,8,9-tetrahydro-5H-benzo[7] wheel En-2-yl)amino)pyrimidin-4-yl)amino)-5-(methoxymethyl)phenyl)dimethylphosphine oxide (compound of formula I) salt and crystal form

Take about 50mg of the compound of formula I, that is, the compound of formula I, and 1.05 equivalents of acid (hydrochloric acid while setting the molar ratio of acid to compound of formula I to 2.10), add 1mL of solvent and stir at room temperature for 2 days. The resulting clear liquid was crystallized by stirring at 5°C and slowly volatilized, and the solid was separated by centrifugation, and then dried by air or under reduced pressure at 40°C for 2-5 hours and then used for XRPD characterization.

Table 15 formula I compound salt formation result

Fig. 4-13 is respectively formula I compound hydrochloride, dihydrochloride, phosphate, hippurate, sulfate, hydrobromide, benzenesulfonate, oxalate, fumarate, citric acid X-ray powder diffraction (XRPD) pattern of the salt.

Example 4 (S)-(2-((5-chloro-2-((7-(pyrrolidin-1-yl)-6,7,8,9-tetrahydro-5H-benzo[7] wheel En-2-yl) amino) pyrimidin-4-yl) amino) -5- (methoxymethyl) phenyl) dimethyl phosphine oxide (formula I compound) methanesulfonate

Add (S)-(2-((5-chloro-2-((7-(pyrrolidin-1-yl)-6,7,8,9-tetrahydro-5H- Benzo[7]annulen-2-yl)amino))pyrimidin-4-yl)amino)-5-(methoxymethyl)phenyl)dimethylphosphine oxide (50 mg), toluene (1 mL) and Methanesulfonic acid (10 mg), stirred and reacted at room temperature for 2 hours to obtain (S)-(2-((5-chloro-2-((7-(pyrrolidin-1-yl)-6 ,7,8,9-tetrahydro-5H-benzo[7]annulen-2-yl)amino))pyrimidin-4-yl)amino)-5-(methoxymethyl)phenyl)dimethyl Phosphine oxide mesylate crystal form.

Example 5 (S)-(2-((5-chloro-2-((7-(pyrrolidin-1-yl)-6,7,8,9-tetrahydro-5H-benzo[7] wheel En-2-yl) amino) pyrimidin-4-yl) amino) -5-(methoxymethyl) phenyl) dimethyl phosphorus oxide (formula I compound) methanesulfonate

Add (S)-(2-((5-chloro-2-((7-(pyrrolidin-1-yl)-6,7,8,9-tetrahydro-5H- Benzo[7]annulen-2-yl)amino))pyrimidin-4-yl)amino)-5-(methoxymethyl)phenyl)dimethylphosphine oxide (1g), ethyl acetate (20mL ) and methanesulfonic acid (172.8 mg), stirred and reacted at room temperature for 10 minutes, and then added the mesylate salt crystal form prepared in Example 4 as a seed crystal (5 mg) and stirred for 30 minutes. Ethyl acetate (20 mL), acetone (10 mL) and the mesylate crystal seed crystal (100. mg) prepared in Example 4 were successively added to the glass vial and stirred for 20 minutes for crystallization. Suction filtration, the wet cake was dried under vacuum at 40°C for 20 hours to obtain (S)-(2-((5-chloro-2-((7-(pyrrolidin-1-yl) )-6,7,8,9-tetrahydro-5H-benzo[7]annulen-2-yl)amino))pyrimidin-4-yl)amino)-5-(methoxymethyl)phenyl ) Dimethylphosphine oxide mesylate crystal form.

¹ H NMR (400MHz, DMSO-d6): 11.11(s,1H); 9.46(br,1H); 9.36(s,1H); 8.58-8.50(m,1H); 8.19(s,1H); 7.43, (m, 3H); 7.37 (dd, J = 8.2, 2.2Hz, 1H); 7.04 (d, J = 8.2Hz, 1H); 4.44 (s, 2H); 3.53-3.46 (m, 3H); 3.33(s,3H); 3.15(s,2H); 2.82-2.62(m,4H); 2.32-2.29(m,5H); 1.99(s,2H); 1.89-1.75(m,8H); 1.40( q,J=12.3Hz,2H).

See accompanying drawing 1 for its XRPD pattern, refer to accompanying drawing 2 for its DSC pattern, and refer to Fig. 3 for its TGA pattern.

Embodiment 6 (S)-(2-((5-chloro-2-((7-(pyrrolidin-1-yl)-6,7,8,9-tetrahydro-5H-benzo[7] wheel Preparation of Form A of En-2-yl)amino)pyrimidin-4-yl)amino)-5-(methoxymethyl)phenyl)dimethylphosphine oxide (compound of formula I)

Add 200 mg of S)-(2-((5-chloro-2-((7-(pyrrolidin-1-yl)-6,7,8,9- Tetrahydro-5H-benzo[7]annulen-2-yl)amino)pyrimidin-4-yl)amino)-5-(methoxymethyl)phenyl)dimethylphosphine oxide and 2 mL of purified water, After 6 hours of magnetic stirring at room temperature, the sample was centrifuged, and the wet sample was dried under reduced pressure at 40°C for 21 hours to obtain 176 mg of Form A, with a yield of 88.0%. The XRPD pattern is shown in Figure 14, X-ray powder diffraction pattern data See Table 16 for details.

Table 16 X-ray powder diffraction pattern data of free base crystal form A

2θ(°)	height (counts)	I/I 0 (%)
4.1	45	15.4
5.6	42	14.3
7.6	109	36.9
10.2	107	36.4
10.9	13	4.4
12.6	59	20.1
13.0	44	14.8
15.2	18	6.2
17.6	246	83.4
19.7	85	28.9
20.3	184	62.5
20.9	294	100.0
22.2	60	20.5
23.2	twenty two	7.5
24.6	19	6.6
27.0	40	13.6
28.8	14	4.6
37.0	6	2.1
37.7	12	4.1

Claims (12)

1. A pharmaceutically acceptable salt of a compound of formula I, the salt is selected from organic acid salts or inorganic acid salts, wherein the organic acid salts are selected from methanesulfonate, benzenesulfonate, oxalate, fumarate A kind of in acid salt, citrate and hippurate, described inorganic acid salt is selected from a kind of in hydrochloride, hydrobromide, sulfate or phosphate, formula I compound structure is as follows:

   
2. According to the pharmaceutically acceptable salt of the compound of formula I according to claim 1, the organic acid salt is mesylate.
3. The pharmaceutically acceptable salt of the compound of formula I according to claim 2, the mesylate salt is in the form of hydrate, further in the form of dihydrate.
4. The pharmaceutically acceptable salt of the compound of formula I according to any one of claims 1-3, wherein the molar ratio of the compound of formula I to the organic acid in the organic acid salt is 1:1.
5. The pharmaceutically acceptable salt of the compound of formula I according to claim 1, wherein the molar ratio of the compound of formula I to the inorganic acid in the inorganic acid salt is 1:1 or 1:2; further, the inorganic acid salt is Hydrochloride, in which the molar ratio of the compound of formula I to hydrogen chloride is 1:1 or 1:2.
6. A crystal form of a pharmaceutically acceptable salt of a compound of formula I, the salt is selected from organic acid salts or inorganic acid salts, wherein the organic acid salts are selected from methanesulfonate, benzenesulfonate, oxalate , fumarate, citrate and hippurate, the inorganic acid salt being selected from hydrochloride, hydrobromide or phosphate.

   
7. The crystal form of the pharmaceutically acceptable salt of the compound of formula I according to claim 6, the salt is a mesylate salt, further, the X-ray powder diffraction of the mesylate salt crystal form of the compound of formula I The picture is shown in Figure 1.
8. A crystal form of a compound of formula I, the X-ray powder diffraction pattern of which has diffraction at 2θ of 7.6°±0.2°, 10.2°±0.2°, 17.6°±0.2°, 20.3°±0.2° and 20.9°±0.2° peak;

   Further, the X-ray powder diffraction pattern is 4.1°±0.2°, 7.6°±0.2°, 10.2°±0.2°, 12.6°±0.2°, 13.0°±0.2°, 17.6°±0.2°, 19.7°±0.2° at 2θ There are diffraction peaks at 0.2°, 20.3°±0.2°, 20.9°±0.2° and 22.2°±0.2°;

   Further, its X-ray powder diffraction pattern at 2θ is 4.1°±0.2°, 5.6°±0.2°, 7.6°±0.2°, 10.2°±0.2°, 10.9°±0.2°, 12.6°±0.2°, 13.0° ±0.2°, 15.2°±0.2°, 17.6°±0.2°, 19.7°±0.2°, 20.3°±0.2°, 20.9°±0.2°, 22.2°±0.2°, 23.2°±0.2°, 24.6°±0.2 °, 27.0°±0.2°, 28.8°±0.2°, 37.0°±0.2° and 37.7°±0.2° have diffraction peaks;

   Further, X-ray powder diffraction represented by 2θ angle has a spectrum as shown in FIG. 14 .
9. The preparation method of the pharmaceutically acceptable salt of the compound of formula I described in any one of claims 1-5 or the crystal form of the pharmaceutically acceptable salt of the compound of formula I described in any one of claims 6-7, It comprises the step of salt-forming a compound of formula I with a corresponding acid.
10. According to the preparation method according to claim 9, the crystal form of the pharmaceutically acceptable salt is a mesylate salt crystal form, and the mesylate salt crystal form is prepared by a method comprising the following steps:

    S1: stirring the compound of formula I, methanesulfonic acid and toluene to obtain a seed crystal of the mesylate salt crystal form of the compound of formula I;

    S2: stirring and reacting the compound of formula (I), methanesulfonic acid and ethyl acetate, adding the crystal form obtained in step S1 and continuing to stir;

    S3: adding ethyl acetate, acetone and the crystal form of the compound of formula I according to claim 8, stirring and crystallizing to obtain the mesylate salt crystal form.
11. A pharmaceutical composition comprising the pharmaceutically acceptable salt of the compound of formula I described in any one of claims 1-5, the pharmaceutically acceptable salt of the compound of formula I described in any one of claims 6-7 Crystal form, the crystal form of the compound of formula I described in claim 8 or the pharmaceutically acceptable salt prepared by the method described in any one of claims 9-10 or the crystal form of the pharmaceutically acceptable salt and the pharmaceutically acceptable salt acceptable carrier.
12. The pharmaceutically acceptable salt of the compound of formula I described in any one of claims 1-5, the crystal form of the pharmaceutically acceptable salt of the compound of formula I described in any one of claims 6-7, claim 8 The crystalline form of the compound of formula I or the pharmaceutically acceptable salt or the crystalline form of the pharmaceutically acceptable salt prepared by the method described in any one of claims 9-10 is used for prevention and/or treatment Use in medicine for an AXL kinase mediated disease or condition.

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