Solid form of heterocyclic-substituted ketone derivative and application thereof

Abstract

The invention relates to a heterocyclic substituted ketone derivative and application thereof. Specifically, the invention provides a solid form of (S)-(3-amino-5-(1-amino-1, 3-dihydrospiro [indene-2, 4 '-piperidine]-1'-yl) pyrazine-2-yl) (2-amino-7, 7-difluoro-6, 7-thiazolo [5, 4-c] pyridine-5 (4H)-yl) ketone and application of the solid form. In addition, the invention also discloses a pharmaceutical composition containing the solid form and application thereof.

Description

Solid forms of heterocyclic substituted ketone derivatives and their applications

[0001] This application claims the priority of the Chinese patent application filed with the China Patent Office on September 22, 2022, with application number 202211159575.9 and invention name “Solid form of heterocyclic substituted ketone derivatives and their applications”, the entire contents of which are incorporated by reference into this application.

[0002] The present invention relates to the field of medical technology, and more specifically, to a solid form of a heterocyclic substituted ketone derivative and its application, wherein the derivative is (S)-(3-amino-5-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)pyrazine-2-yl)(2-amino-7,7-difluoro-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-yl)ketone.

[0003] Protein tyrosine phosphatases (PTPs) catalyze the dephosphorylation of phosphotyrosine and are key control elements in mammalian signal transduction. Deviations in their biological functions can cause regulatory disorders in the body and lead to the occurrence of a variety of diseases such as cancer, diabetes, and autoimmune diseases. Among them, Src homology 2 domain-containing protein tyrosine phosphatase (SHP2) is currently the only confirmed proto-oncoprotein in the PTP family. Src homology-2 phosphatase (SHP2) is a non-receptor protein tyrosine phosphatase encoded by the PTPN11 gene that contributes to a variety of cellular functions including proliferation, differentiation, cell cycle maintenance, and migration. SHP2 is involved in signal transduction through the Ras-mitogen-activated protein kinase (JAK-STAT or phosphoinositide 3-kinase-AKT pathway).

[0004] Mutations in the PTPN11 gene and subsequently SHP2 have been identified in a variety of human diseases, including, for example, Noonan syndrome, Leopard syndrome, juvenile myelomonocytic leukemia, neuroblastoma, melanoma, acute myeloid leukemia, and breast, lung and colon cancers. Therefore, SHP2 is a highly attractive target for the development of new therapies for the treatment of a variety of diseases. The compounds of formula (I) of the present invention meet the need for small molecules that inhibit SHP2 activity.

[0005] The stability and other properties of the active ingredients of drugs have an important impact on the drugability research, and the solid form is conducive to the quality control of the product. Therefore, it is necessary to explore the solid form of the product.

[0006]

[0007] Based on this, the object of the present invention is to provide a new solid form and application of heterocyclic substituted ketone derivatives.

[0008] The first aspect of the present invention provides a solid form of a compound of formula (I):

[0009] The solid forms are polymorphs.

[0010] In some embodiments, the polymorph is Form I, and its X-ray powder diffraction (XRPD) pattern 2θ (°) diffraction angle has characteristic diffraction peaks at values ​​of 15.982±0.2, 20.367±0.2 and 21.114±0.2.

[0011] In some embodiments, the XRPD pattern 2θ (°) diffraction angle of the Form I has characteristic diffraction peaks at values ​​of 15.982±0.2, 20.367±0.2 and 21.114±0.2, and at least one value selected from 8±0.2, 9.232±0.2 and 13.586±0.2.

[0012] In some embodiments, the XRPD pattern 2θ (°) diffraction angle of the crystalline Form I has characteristic diffraction peaks at values ​​of 15.982±0.2, 20.367±0.2 and 21.114±0.2, and at least one value selected from 8±0.2, 9.232±0.2, 11.088±0.2, 12.438±0.2, 13.586±0.2, 17.181±0.2 and 19.523±0.2.

[0013] In some embodiments, the XRPD pattern 2θ (°) diffraction angle of the crystalline Form I has characteristic diffraction peaks at values ​​of 15.982±0.2, 20.367±0.2 and 21.114±0.2, and at least one value selected from 8±0.2, 9.232±0.2, 11.088±0.2, 12.438±0.2, 13.315±0.2, 13.586±0.2, 14.598±0.2, 15.025±0.2, 17.181±0.2, 19.523±0.2 and 19.943±0.2.

[0014] In some embodiments, the XRPD pattern 2θ (°) diffraction angle of Form I is at 15.982±0.2, 20.367±0.2 and 21.114±0.2 values, and is selected from 8±0.2, 9.232±0.2, 11.088±0.2, 12.438±0.2, 13.315±0.2, 13.586±0.2, 14.598±0 .2, 15.025±0.2, 17.181±0.2, 19.523±0.2, 19.943±0.2, 22.19±0.2, 23.24±0.2, 24.776±0.2, 25.402±0.2, 26.064±0.2, 27.082±0.2 and 28.101±0.2 have a characteristic diffraction peak at at least one value.

[0015] In some embodiments, the XRPD pattern 2θ (°) diffraction angle of Form I is at 15.982±0.2, 20.367±0.2 and 21.114±0.2 values, and 1 or more (such as 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or 21) selected from 8±0.2, 9.232±0.2, 11.088±0.2, 12.438±0.2, 13.315±0.2, 13 The diffraction peaks are shown at 0.586±0.2, 14.598±0.2, 15.025±0.2, 17.181±0.2, 18.168±0.2, 19.523±0.2, 19.943±0.2, 22.19±0.2, 23.24±0.2, 24.776±0.2, 25.402±0.2, 26.064±0.2, 27.082±0.2, 28.101±0.2, 29.505±0.2 and 34.098±0.2.

[0016] In some embodiments, the XRPD pattern 2θ (°) diffraction angle of the crystalline form I has characteristic diffraction peaks at at least three values ​​selected from the following diffraction angles 2θ (°): 8±0.2, 9.232±0.2, 11.088±0.2, 12.438±0.2, 13.315±0.2, 13.586±0.2, 14.598±0.2, 15.025±0.2, 1 5.982±0.2, 17.181±0.2, 19.523±0.2, 19.943±0.2, 20.367±0.2, 21.114±0.2, 22.19±0.2, 23.24±0.2, 24.776±0.2, 25.402±0.2, 26.064±0.2, 27.082±0.2 and 28.101±0.2.

[0017] In some embodiments, the XRPD pattern 2θ (°) diffraction angle of the crystalline form I has characteristic diffraction peaks at 3 or more (such as 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24) or all selected from the following diffraction angle 2θ (°) values: 8 ± 0.2, 9.232 ± 0.2, 11.088 ± 0.2, 12.438 ± 0.2, 13.315 ± 0.2, 13.586 ± 0.2, 14.598 ± 0 .2, 15.025±0.2, 15.982±0.2, 17.181±0.2, 18.168±0.2, 19.523±0.2, 19.943±0.2, 20.367±0.2, 21.114±0.2, 22.19±0.2, 23.24±0.2, 24.776±0.2, 25.402±0.2, 26.064±0.2, 27.082±0.2, 28.101±0.2, 29.505±0.2 and 34.098±0.2.

[0018] In some embodiments, the XRPD pattern 2θ (°) diffraction angle of Form I has characteristic diffraction peaks at 8±0.2, 9.232±0.2, 13.586±0.2, 15.982±0.2, 20.367±0.2 and 21.114±0.2.

[0019] In some embodiments, the XRPD pattern 2θ (°) diffraction angle of the Form I has characteristic diffraction peaks at 8±0.2, 9.232±0.2, 11.088±0.2, 12.438±0.2, 13.586±0.2, 15.982±0.2, 17.181±0.2, 19.523±0.2, 20.367±0.2 and 21.114±0.2.

[0020] In some embodiments, the XRPD pattern 2θ (°) diffraction angle of the crystalline Form I has characteristic diffraction peaks at 8±0.2, 9.232±0.2, 11.088±0.2, 12.438±0.2, 13.315±0.2, 13.586±0.2, 14.598±0.2, 15.025±0.2, 15.982±0.2, 17.181±0.2, 19.523±0.2, 19.943±0.2, 20.367±0.2 and 21.114±0.2 values.

[0021] In some embodiments, the XRPD pattern 2θ (°) diffraction angle of the crystalline Form I has characteristic diffraction peaks at 9.232±0.2, 12.438±0.2, 13.315±0.2, 13.586±0.2, 14.598±0.2, 15.025±0.2, 15.982±0.2, 17.181±0.2, 19.523±0.2, 19.943±0.2, 20.367±0.2, 21.114±0.2, 23.24±0.2, 24.776±0.2, 25.402±0.2, 26.064±0.2 and 28.101±0.2 values.

[0022] In some embodiments, the XRPD pattern 2θ (°) diffraction angle of Form I is 8±0.2, 9.232±0.2, 11.088±0.2, 12.438±0.2, 13.315±0.2, 13.586±0.2, 14.598±0.2, 15.025±0.2, 15.982±0.2, 17.181±0 .2, 19.523±0.2, 19.943±0.2, 20.367±0.2, 21.114±0.2, 22.19±0.2, 23.24±0.2, 24.776±0.2, 25.402±0.2, 26.064±0.2, 27.082±0.2 and 28.101±0.2, with characteristic diffraction peaks.

[0023] In some embodiments, the XRPD pattern 2θ (°) diffraction angle of the crystalline form I is 8±0.2, 9.232±0.2, 11.088±0.2, 12.438±0.2, 13.315±0.2, 13.586±0.2, 14.598±0.2, 15.025±0.2, 15.982±0.2, 17.181±0.2, 18.168±0.2, 19. There are characteristic diffraction peaks at 523±0.2, 19.943±0.2, 20.367±0.2, 21.114±0.2, 22.19±0.2, 23.24±0.2, 24.776±0.2, 25.402±0.2, 26.064±0.2, 27.082±0.2, 28.101±0.2, 29.505±0.2 and 34.098±0.2.

[0024] In some embodiments, the X-ray powder diffraction pattern of the crystalline form I expressed in terms of 2θ (°) value and interplanar spacing (d) value has characteristic diffraction peaks shown in Table 1, and the relative intensities of the peaks are shown in Table 1, wherein the diffraction peak with the highest peak height is taken as the base peak, and its relative intensity is defined as 100%, as I0 (the peak with a 2θ (°) value of 21.114 of the crystalline form I is the base peak), and the ratio of the peak height to the base peak height of the other peaks is taken as their relative intensity I / I0:

[0025] Table 1 2θ(°), d value and relative intensity I / I0 of Form I

[0026] In some embodiments, the XRPD pattern of Form I is substantially as shown in Figure 2.

[0027] In some embodiments, the thermogravimetric analysis spectrum (TGA spectrum) of the crystalline form I is substantially as shown in FIG. 3 .

[0028] In some embodiments, the TGA spectrum of the crystalline Form I shows a weight loss of no more than 0.6% between 0-120°C.

[0029] In some embodiments, the TGA spectrum of the crystalline Form I shows a weight loss of 0.559% at around 120°C.

[0030] In some embodiments, the crystalline Form I is an anhydrate.

[0031] The second aspect of the present invention provides a method for preparing a solid form of the compound of formula (I) described in the first aspect, comprising the following steps:

[0032] (a) dispersing the free base of the compound of formula (I) in a solvent to obtain a liquid;

[0033] (b) The liquid in step (a) is subjected to crystallization to obtain a solid form of the compound of formula (I).

[0034] In some embodiments, the solid form of the compound of formula (I) is Form I.

[0035] In some embodiments, the free base of the compound of formula (I) described in step (a) is in an amorphous form.

[0036] In some embodiments, the mass volume ratio (w / v, in mg / mL) of the free base of the compound of formula (I) to the solvent in step (a) is 1:1-20:1; preferably 2:1-18:1; more preferably 3:1-16:1.

[0037] In some embodiments, the mass volume ratio (w / v, in mg / mL) of the free base of the compound of formula (I) to the solvent in step (a) is 3:1 or 15:1.

[0038] In some embodiments, the solvent in step (a) is selected from one or more of methanol, ethanol, isopropanol, acetonitrile, ethyl acetate, dichloromethane, dichlorohexane, n-heptane, n-hexane, methyl tert-butyl ether, isopropyl ether, acetone, 2-butanone, purified water, toluene, and tetrahydrofuran; preferably, selected from one or more of ethanol, ethyl acetate, dichlorohexane, n-heptane, and methyl tert-butyl ether.

[0039] In some embodiments, the liquid in step (a) is a solution or a suspension.

[0040] In some embodiments, the crystallization treatment in step (b) is selected from suspension shaking, stirring, suspension centrifugation, slow volatilization, cooling crystallization, and / or adding anti-solvent. In some embodiments, the crystallization treatment in step (b) is stirring, and / or cooling crystallization.

[0041] In some embodiments, the temperature of the crystallization treatment in step (b) is 0-80°C, for example, it can be any range consisting of end values ​​selected from 0°C, 5°C, 10°C, 15°C, 20°C, 25°C, 30°C, 35°C, 40°C, 45°C, 50°C, 55°C, 60°C, 65°C, 70°C, 75°C and 80°C, without specific limitation.

[0042] In some embodiments, the crystallization treatment time in step (b) is 0.25-10 days, for example, it can be any range consisting of end values ​​selected from 0.25, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5 and 10, without specific limitation.

[0043] In some embodiments, step (b) further comprises separation, and / or drying steps.

[0044] In some embodiments, the separation is separation by filtration, centrifugation or a combination thereof.

[0045] In some embodiments, the drying is vacuum drying or evaporation of the solvent.

[0046] In some embodiments, the method for preparing the crystalline form I comprises the following steps:

[0047] a. dispersing the amorphous form of the compound of formula (I) in a solvent to obtain a liquid;

[0048] b. The liquid in step (a) is stirred at 10-35°C for 2-5 days, and the solid is collected to obtain Form I of the compound of formula (I).

[0049] In some embodiments, the solvent in step (a) is selected from one or more of methanol, ethanol, acetonitrile, acetone, 2-butanone, isopropyl ether, purified water, dichloromethane, methyl tert-butyl ether, toluene, n-heptane, ethyl acetate, tetrahydrofuran, and isopropanol.

[0050] In some embodiments, the solvent in step (a) is selected from one or more of methanol, acetonitrile, ethyl acetate, dichloromethane, n-heptane, ethanol, and methyl tert-butyl ether.

[0051] In some embodiments, the solvent in step (a) is selected from acetonitrile or ethanol.

[0052] In some embodiments, the temperature in step (b) is 15-30°C, for example, it can be any range consisting of end values ​​selected from 15°C, 16°C, 17°C, 18°C, 19°C, 20°C, 21°C, 22°C, 23°C, 24°C, 25°C, 26°C, 27°C, 28°C, 29°C and 30°C, without specific limitation.

[0053] In some embodiments, the temperature in step (b) is 10-25°C, for example, it can be any range consisting of end values ​​selected from 10°C, 11°C, 12°C, 13°C, 14°C, 15°C, 16°C, 17°C, 18°C, 19°C, 20°C, 21°C, 22°C, 23°C, 24°C and 25°C, without specific limitation.

[0054] In some embodiments, the time described in step (b) is 2.5-5 days, for example, it can be any range consisting of end values ​​selected from 2.5 days, 3 days, 3.5 days, 4 days, 4.5 days and 5 days, without specific limitation.

[0055] In some embodiments, step (b) collects the solids by solid-liquid separation.

[0056] In some embodiments, the separation is separation by filtration, centrifugation or a combination thereof.

[0057] In some embodiments, step (b) further comprises a drying step after collecting the solid.

[0058] In some embodiments, the drying is vacuum drying or evaporation of the solvent.

[0059] In some embodiments, the method for preparing the crystalline form I comprises the following steps:

[0060] a. dispersing the amorphous form of the compound of formula (I) in a solvent to obtain a liquid;

[0061] b. The liquid in step (a) is stirred at a first temperature for a first period of time, cooled to a second temperature, and then stirred at the second temperature for a second period of time to obtain Form I of the compound of formula (I).

[0062] In some embodiments, the solvent in step (a) is selected from one or more of ethanol, ethyl acetate, dichloromethane, n-heptane, and methyl tert-butyl ether.

[0063] In some embodiments, the first temperature described in step (b) is 45-70°C, for example, it can be any range consisting of end values ​​selected from 45°C, 46°C, 47°C, 48°C, 49°C, 50°C, 51°C, 52°C, 53°C, 54°C, 55°C, 56°C, 57°C, 58°C, 59°C, 60°C, 61°C, 62°C, 63°C, 64°C, 65°C, 66°C, 67°C, 68°C, 69°C and 70°C, without specific limitation.

[0064] In some embodiments, the first time period in step (b) is 2-6 hours, for example, it can be any range consisting of end values ​​selected from 2 hours, 3 hours, 4 hours, 5 hours and 6 hours, without specific limitation.

[0065] In some embodiments, the second temperature of step (b) is 10-40°C, for example, it can be any range consisting of end values ​​selected from 10°C, 11°C, 12°C, 13°C, 14°C, 15°C, 16°C, 17°C, 18°C, 19°C, 20°C, 21°C, 22°C, 23°C, 24°C, 25°C, 26°C, 27°C, 28°C, 29°C, 30°C, 31°C, 32°C, 33°C, 34°C, 35°C, 36°C, 37°C, 38°C, 39°C and 40°C, without specific limitation.

[0066] In some embodiments, the second period of time in step (b) is 0.25-3 days, for example, any range consisting of end values ​​selected from 0.25 days, 0.5 days, 1 day, 1.5 days, 2 days, 2.5 days and 3 days, without specific limitation.

[0067] The third aspect of the present invention provides a pharmaceutical composition, which comprises: a solid form of the compound of formula (I) according to the first aspect of the present invention, and a pharmaceutically acceptable carrier.

[0068] In some embodiments, the pharmaceutical composition comprises Form I of the compound of formula (I), and a pharmaceutically acceptable carrier.

[0069] The fourth aspect of the present invention provides the use of a solid form of a compound of formula (I) as described in the first aspect of the present invention or a pharmaceutical composition as described in the third aspect of the present invention in the preparation of a medicament for treating and / or preventing diseases or conditions mediated by SHP2 or associated with abnormal SHP2 activity.

[0070] The fifth aspect of the present invention provides a method for treating a disease or condition mediated by SHP2 or associated with abnormal SHP2 activity, the method comprising administering to a subject a therapeutically effective amount of a solid form of a compound of formula (I) as described in the first aspect of the present invention or a pharmaceutical composition as described in the third aspect of the present invention.

[0071] Src homology-2 phosphatase (SHP2) is a protein tyrosine phosphatase encoded by the PTPN11 gene that contributes to a variety of cellular functions including proliferation, differentiation, cell cycle maintenance and migration. SHP2 is involved in signal transduction through the Ras-mitogen activated protein kinase, i.e., the JAK-STAT or phosphoinositide 3-kinase-AKT pathway. SHP2 mediates the activation of extracellular regulated protein kinase (Erk) 1 and Erk2 (Erk1 / 2, Erk) mitogen activated protein kinases (MAP kinases) through receptor tyrosine kinases such as epidermal growth factor receptor (ErbB) 1, ErbB2 and hepatocyte growth factor receptor (c-Met).

[0072] SHP2 has two N-terminal Src homology 2 domains (N-SH2 and C-SH2), a protein tyrosine phosphatase (PTP) catalytic domain and a C-terminal tail. The two SH2 domains control the subcellular localization and functional regulation of SHP2. The compound molecule exists in an inactive conformation and inhibits its own activity through a binding network involving residues from both the N-SH2 and PTP domains. In response to growth factor stimulation, SHP2 binds to specific tyrosine-phosphorylated sites on docking proteins such as cell phosphorylation signal adapter molecules (Gab) 1 and Gab2 through its SH2 domain. This induces conformational changes that lead to SHP2 activation.

[0073] In some embodiments, diseases or conditions associated with abnormal SHP2 activity include: solid tumors and hematological tumors. In some embodiments, the SHP2-mediated disease or condition is cancer, including but not limited to: juvenile myelomonocytic leukemia (JMML), acute myeloid leukemia (AML), B-cell acute lymphoblastic leukemia (B-ALL), neuroblastoma, esophageal cancer, breast cancer, lung cancer, colon cancer, gastric cancer, head and neck cancer.

[0074] The crystalline form I of the compound of formula (I) provided by the present invention has good stability at various temperatures (room temperature and high temperature) and in various solvents, which is more conducive to the development of preparations.

[0075] The drawings described herein are used to provide further understanding of the present invention and constitute a part of the present invention. The exemplary embodiments of the present invention and their descriptions are used to explain the present invention and do not constitute improper limitations on the present invention.

[0076] FIG1 is a single crystal structure diagram of compound 5;

[0077] FIG2 is an XRPD pattern of Form I of the compound of formula (I);

[0078] FIG3 is a TGA spectrum of the crystalline form I of the compound of formula (I);

[0079] FIG4 is the XRPD spectrum of the compound of formula (I) obtained in Preparation Example 2.

[0080] In order to make the purpose, technical scheme and advantages of the present invention more clearly understood, the present invention is further described in detail with reference to the accompanying drawings and embodiments. Obviously, the described embodiments are only part of the embodiments of the present invention, rather than all the embodiments. All other embodiments obtained by those skilled in the art based on the present invention belong to the scope of protection of the present invention.

[0081] In the present invention, the compound of formula (I) is (S)-(3-amino-5-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl)pyrazin-2-yl)(2-amino-7,7-difluoro-6,7-dihydrothiazolo[5,4-c]pyridine-5(4H)-yl)methanone, which has the following structure:

[0082] The solid form of the compound of formula (I) of the present invention is a polymorph, including the crystalline form I of the compound of formula (I).

[0083] Polymorphs: Solids exist in either amorphous form or in crystalline form. In the case of crystalline form, the molecules are positioned in a three-dimensional lattice. When a compound crystallizes from a solution or slurry, it can crystallize in different spatial arrangements (a property known as "polymorphism"), forming crystals with different crystalline forms, which are known as "polymorphs". Different polymorphs of a given substance may differ from one another in one or more physical properties, such as solubility and dissolution rate, true specific gravity, crystal shape, packing pattern, flowability and / or solid state stability.

[0084] Crystallization: Crystallization on a production scale can be accomplished by manipulating the solution so that the solubility limit of the compound of interest is exceeded. This can be accomplished in a variety of ways, for example, by dissolving the compound at a relatively high temperature and then cooling the solution to below the saturation limit. Alternatively, the liquid volume can be reduced by boiling, atmospheric evaporation, vacuum drying, or by some other method. The solubility of the compound of interest can be reduced by adding an antisolvent or a solvent or a mixture of such solvents in which the compound has a low solubility. Another alternative is to adjust the pH to reduce the solubility.

[0085] The "suspension centrifugation" of the present invention refers to a method of mixing the compound of formula (I) in a suitable solvent to form a turbid solution and then centrifuging to obtain crystals. The suitable solvent may be water or an organic solvent.

[0086] Optimization of crystallization can include seeding the crystallization medium with crystals of the desired form. In addition, many crystallization methods use a combination of the above strategies. One embodiment dissolves the compound of interest in a solvent at high temperature, followed by the addition of an appropriate volume of antisolvent in a controlled manner to bring the system just below the saturation level. At this point, seeds of the desired form can be added (while maintaining the integrity of the seeds), and the system is cooled to complete crystallization. As used herein, the term "about" means ±5 on the basis of a given value.

[0087] In this article, when referring to the unit of a data range, if there is a unit only after the right endpoint, it means that the units of the left and right endpoints are the same. For example, 3~5h means that the units of the left endpoint "3" and the right endpoint "5" are both h (hours).

[0088] Herein, "preferred", "better", "more preferred" and "suitable" are only used to describe implementation methods or examples with better effects, and it should be understood that they do not constitute a limitation on the scope of protection of the present invention. If multiple "preferred" items appear in a technical solution, unless otherwise specified and there is no contradiction or mutual restriction, each "preferred" item is independent.

[0089] The terms "and / or", "or / and", and "and / or" used herein include any one of two or more related listed items, and also include any and all combinations of the related listed items, wherein the any and all combinations include any combination of two related listed items, any more related listed items, or all related listed items. It should be noted that when at least three items are connected by at least two conjunctions selected from "and / or", "or / and", and "and / or", it should be understood that in the present invention, the technical solution undoubtedly includes a technical solution connected by "logical and", and also undoubtedly includes a technical solution connected by "logical or".

[0090] As used herein, "plurality", "multiple", "multiple times", etc., unless otherwise specified, refer to a number greater than or equal to 2. For example, "multiple" means greater than or equal to two.

[0091] In this article, the technical features described in an open manner include closed technical solutions composed of the listed features, and also include open technical solutions containing the listed features.

[0092] As used herein, the term "solid form of the present invention" or "solid form of a compound of Formula (I) of the present invention" includes, but is not limited to, Form I of the compound of Formula (I).

[0093] “Compound of formula (I)”, “free base of compound of formula (I)” and “free base” can be used interchangeably, and “polymorph of compound of formula (I)” and “polymorph of free base of compound of formula (I)” can be used interchangeably.

[0094] "Form I" and "Form I of the compound of formula (I)" are used interchangeably.

[0095] Pharmaceutical composition and its application

[0096] Usually, the solid form of the compound of formula (I) of the present invention as an active ingredient can be administered in a suitable dosage form together with one or more pharmaceutically acceptable carriers.

[0097] "Pharmaceutically acceptable carrier" refers to a non-toxic, inert, solid, semi-solid substance or liquid filler, diluent, encapsulating material or auxiliary formulation or any type of excipient, which is compatible with the subject to be administered (preferably a mammal, more preferably a human), and is suitable for delivering the active substance of the present invention to the target site without terminating its activity.

[0098] The pharmaceutical composition of the present invention is formulated, dosed and administered in a manner consistent with medical practice. The "therapeutically effective amount" of the active ingredient administered is determined by factors such as the specific condition to be treated, the individual being treated, the cause of the condition, the target of the drug, and the mode of administration.

[0099] The present invention provides a solid form of the compound of formula (I) according to the first aspect of the present invention and a pharmaceutical composition according to the third aspect of the present invention which can be used to prepare a drug for treating and / or preventing diseases or conditions mediated by SHP2 or associated with abnormal SHP2 activity.

[0100] The present invention provides a method for treating a disease or condition mediated by SHP2 or associated with abnormal SHP2 activity, comprising administering to a subject a therapeutically effective amount of a solid form of a compound of formula (I) according to the first aspect of the present invention, or a pharmaceutical composition according to the third aspect of the present invention.

[0101] As used herein, "therapeutically effective amount" refers to the amount of a solid form of a compound of formula (I) of the present invention that will elicit a biological or medical response in a subject, such as reducing or inhibiting enzyme or protein activity or improving symptoms, alleviating symptoms, relieving or delaying disease progression, or preventing disease, etc.

[0102] As used herein, "subject" refers to an animal, preferably a mammal, more preferably a human. The term "mammal" refers to warm-blooded vertebrate mammals, including cats, dogs, rabbits, bears, foxes, wolves, monkeys, deer, mice, pigs and humans.

[0103] "Treatment" means to lessen, slow the progression, attenuate, prevent, or maintain an existing disease or condition (eg, cancer). Treatment also includes curing, preventing the development of, or alleviating to some extent, one or more symptoms of a disease or condition.

[0104] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental methods in the following examples that do not specify specific conditions are usually based on conventional conditions or the conditions recommended by the manufacturer. Unless otherwise specified, percentages and parts are calculated by weight. Unless otherwise defined, the terms used herein have the same meaning as those familiar to those skilled in the art. In addition, any method and material similar or equivalent to the described content can be applied to the present invention.

[0105] Reagents and instruments

[0106] In the present invention, the structure and purity of the compound are determined by nuclear magnetic resonance ( 1 H-NMR) and / or liquid chromatography-mass spectrometry (LC-MS).

[0107] 1 H-NMR: Bruker AVANCE-400 nuclear magnetometer, internal standard is tetramethylsilane (TMS).

[0108] LC-MS: Agilent 1290 HPLC System / 6130 / 6150 MS liquid chromatography-mass spectrometer (manufacturer: Agilent), column: Waters BEH / CHS, 50×2.1 mm, 1.7 μm.

[0109] High performance liquid chromatography (HPLC) analysis was performed using Agilent 1260 Infinity HPLC, OpenLAB CDS Chemstation workstation, XBridge C18 4.6*250mm, ID 5μm column, and detector DAD.

[0110] Elemental analysis was performed using an inductively coupled plasma emission spectrometer, model icp 500, power 1300 w, flow rate 1 mL / min.

[0111] Known starting materials can be used or synthesized according to methods known in the art, or can be purchased from companies such as ABCR GmbH & Co. KG, Acros Organics, Aldrich Chemical Company, Accela ChemBio Inc, and Darui Chemicals.

[0112] As used herein, room temperature in the following examples refers to about 20-30°C, and overnight refers to about 10 h to 16 h.

[0113] Yield = actual product mass / theoretical product mass × 100%.

[0114] General Methods

[0115] X-ray powder diffraction (XRPD): In the present invention, the X-ray diffraction pattern of the above-mentioned crystalline or amorphous powder is obtained by a known method in the art using an ARL Equinox3000 X-ray powder diffraction analyzer. The XRPD test parameters are shown in Table 2 below:

[0116] Table 2

[0117] In the powder X-ray diffraction pattern, the position of each peak is determined by the 2θ (°) value. It is understood that different instruments and / or conditions may result in slightly different data, and the position and relative intensity of each peak may vary.

[0118] Single crystal X-ray diffraction (SXRD): In the present invention, the single crystal X-ray diffraction pattern of compound 5 is obtained by a known method in the art using a D8 Venture X-ray diffractometer, and the SXRD test parameters are shown in Table 3. After collecting relevant data, the crystal structure is further analyzed using a direct method (SHELXT2014) to confirm the absolute configuration.

[0119] Table 3

[0120] High performance liquid chromatography: In the present invention, high performance liquid chromatography (HPLC) was collected on an Agilent 1260 HPLC.

[0121] Thermogravimetric analysis (TGA): In the present invention, the thermogravimetric analysis spectrum of the above-mentioned crystal form is obtained by a known method in the art using a TGA550 thermogravimetric analyzer. The TGA test parameters are shown in Table 4 below:

[0122] Table 4

[0123] It is understood that other types of instruments having the same function as the above-mentioned instruments or using different test conditions than those used in the present invention may result in other values, and therefore, the quoted values ​​should not be regarded as absolute values.

[0124] Due to instrument errors or differences in operators, those skilled in the art will understand that the above parameters used to characterize the physical properties of the crystals may have slight differences, so the above parameters are only used to assist in characterizing the polymorphs provided by the present invention and cannot be regarded as limitations on the polymorphs of the present invention.

[0125] The MTBE used in the present invention refers to methyl tert-butyl ether; P(tBu)3Pd G2: chloro[(tri-tert-butylphosphine)-2-(2-aminobiphenyl)]palladium(II); DMF: N,N-dimethylformamide; THF refers to tetrahydrofuran; LDA: lithium diisopropylamide; PE: petroleum ether; EA: ethyl acetate; Ti(OEt)4: tetrabutyl titanate; DIBAL-H: diisobutylaluminum hydride; DCM: dichloromethane; MeOH: methanol; HATU: 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate; TEA: triethylamine; DMSO: dimethyl sulfoxide; N-Boc-4-cyanopiperidine: N-tert-butyloxycarbonyl-4-cyanopiperidine; DIPEA: diisopropylethylamine.

[0126] Preparation Example 1: Preparation of Compound 6

[0127] Step 1: Dissolve N-Boc-4-cyanopiperidine (648 mg, 3.08 mmol) in THF (10 mL), slowly add LDA (373.03 mg, 3.48 mmol, 1.74 mL) under N2 protection at -78 ° C, stir and react for 45 minutes to obtain a reaction solution. Dissolve compound 1 (3.70 mmol) in THF (3 mL) and slowly dropwise add it to the reaction solution. After the addition is completed, continue to stir and react at 25 ° C for 45 minutes. Monitor the reaction by LC-MS and thin layer chromatography (TLC, PE / EA=10 / 1) until the reaction is completed. The reaction mixture is purified by silica gel column chromatography (12 g silica gel, 0-14% EA / 100-86% PE) to obtain compound 2.

[0128] Step 2: Compound 2 (1.34 mmol) was dissolved in DMF (5 mL) and water (500 μL), and P(tBu)3Pd G2 (138 mg, 269.32 μmol) and triethylamine (326.48 mg, 3.23 mmol, 450 μL) were added under N2 protection, and the mixture was heated and stirred in an oil bath at 130°C for 12 hours. After the reaction was cooled to room temperature, 150 mL of water was added to quench the reaction, and the mixture was poured into a separatory funnel, and ethyl acetate (30 mL×2) was added for extraction. The organic phases were combined, washed with saturated brine (40 mL), dried over anhydrous sodium sulfate, filtered, concentrated and dried. Purification was performed on a silica gel column (4 g silica gel, 0-16% EA / 100-84% PE) to obtain compound 3.

[0129] Step 3: Compound 3 (1.20 mmol), (R)-(+)-tert-butylsulfenamide (219.00 mg, 1.81 mmol) and Ti(OEt)4 (882 mg, 4.82 mmol, 1.6 mL) were placed in a reaction flask, and reacted at 80°C under N2 protection. The reaction was monitored by LC-MS and TLC (PE / EA=5 / 1). The raw material disappeared and the reaction was completed. After the reaction was cooled to room temperature, water was added to quench the reaction, ethyl acetate (30 mL) was added to dissolve, filtered through a diatomaceous earth layer, and the filtrate was poured into a separatory funnel, extracted with ethyl acetate (30 mL×2), and the organic phases were combined, washed with saturated brine (40 mL), dried over anhydrous sodium sulfate, filtered, concentrated and dried. Purification by silica gel column (12 g silica gel, 0-21% EA / 100-79% PE) gave compound 4.

[0130] Step 4: Compound 4 (929.39 μmol) was dissolved in THF (5.0 mL) and DIBAL-H (199.50 mg, 1.41 mmol, 250 μL) was added dropwise, and the reaction was stirred for 2 h. The reaction was monitored by LC-MS and TLC (PE / EA=5 / 1) until the reaction was completed. An appropriate amount of water was added to quench the reaction, filtered through a diatomaceous earth layer, the filter cake was washed with ethyl acetate (40 mL), the filtrate was washed with saturated brine (40 mL), the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated and dried. Purification by silica gel column (4 g silica gel, 0-32% EA / 100-68% PE) gave compound 5.

[0131] Step 5: Dissolve compound 5 (533.72 μmol) in DCM (3 mL), add trifluoroacetic acid (471 mg, 4.13 mmol) dropwise, and stir to react for 2 h. Monitor the reaction by LC-MS and TLC (DCM / MeOH=10 / 1) until the reaction is complete. The reaction solution is dried under reduced pressure, DCM (10 mL) is added, and then concentrated and dried. Purify on a silica gel column (4 g silica gel, 0-20% MeOH / 100-80% DCM) to obtain compound 6.

[0132] Confirmation of the absolute configuration of compound 6

[0133] Add 100 mg of compound 5, 1 mL of ethyl acetate, and 5 mL of heptane to a white transparent glass bottle, shake until dissolved, filter, evaporate slowly, and precipitate crystals. The crystals were subjected to single crystal X-ray (X-ray) analysis using a Bruker D8 venture X-ray single crystal diffractometer, confirming that its absolute configuration is (S) configuration, and its structure is shown in Figure 1. Therefore, the absolute configuration of compound 5 is (S) configuration, and the absolute configuration of compound 6 is also (S) configuration. That is, the structure of compound 6 is as follows:

[0134] Preparation Example 2: Preparation of the compound of formula (I)

[0135] Step 1: Dissolve compound I-1 (2.35 g, 10 mmol), cyanamide (840 mg, 20 mmol), and sulfur powder (640 mg, 20 mmol) in pyridine (15 mL). The reaction was stirred at 130 ° C for 2 hours. After the reaction was completed, it was cooled to room temperature, 60 mL of ethyl acetate was added, and diatomaceous earth was filtered. The filter cake was washed with 50 mL of ethyl acetate 3 times, and the organic phase was decompressed and dried to remove the solvent. Compound I-2 (3 g, 100% yield) was obtained. MS m / z (ESI): 292.1 [M+1] + .

[0136] Step 2: Dissolve compound I-2 (3 g, 10.3 mmol) in MeOH (40 mL), and then add HCl in dioxane (4M (mol / L), 25 mL). The reaction was stirred at 25 ° C for 2 hours. After the reaction, the solvent was dried under reduced pressure. Alkalize with DIPEA, and purify by column separation (20 g silica gel, 1-12% MeOH / 99-88% DCM) to obtain compound I-3 (2.5 g, 100% yield). MS m / z (ESI): 192.0 [M+1] + .

[0137] Step 3: Dissolve 3-amino-5-chloropyrazine-2-carboxylic acid (173 mg, 1 mmol) and compound I-3 (573 mg, 3 mmol) in ultra-dry DMF (10 mL), add HATU (570 mg, 1.5 mmol), TEA (131 mg, 1.3 mmol), and stir at room temperature for 1 hour. Add 50 mL of water, extract with ethyl acetate (30 mL × 3), wash the organic phase with water (50 mL × 2) and saturated brine (50 mL), add anhydrous sodium sulfate (5 g) to the organic phase, stir for 5 minutes, filter to obtain the filtrate, and concentrate under reduced pressure to obtain compound I-4 (303 mg, 87.57% yield). MS m / z (ESI): 347.0 [M+1] + .

[0138] Step 4: Dissolve compound I-4 (70 mg, 0.2 mmol) and compound 6 (73 mg, 0.24 mmol) in ultra-dry DMF (5 mL), add potassium carbonate (83 mg, 0.6 mmol), and stir at 70 ° C for 3 hours. After concentration under reduced pressure, compound I-5 (59 mg 47.9% yield) was obtained after column separation (4 g silica gel, 1-10% MeOH / 99-90% DCM) purification. MS m / z (ESI): 617.2 [M+1] + .

[0139] Step 5: Dissolve compound I-5 (59 mg, 0.096 mmol) in MeOH (3 mL), add dioxane hydrochloride solution (4M (mol / L), 0.3 mL) to the reaction solution, and stir the reaction at room temperature (24°C) for 20 minutes. Separate and purify by preparative liquid phase to obtain compound of formula (I) (21 mg, 42.85% yield). MS m / z (ESI): 513.2 [M+1] + . 1 H-NMR (400MHz, DMSO-d6): δ7.58 (s, 1H), 7.28 (d, J = 6.0 Hz, 1H), 7.22-7.09 (m, 5H), 6.77 (s, 2H), 4.81 (s, 2H), 4.45 (s, 2H), 4.28-4.22 (m, 2H), 3.82 (s, 1H), 3.15-3.03 (m, 3H), 2.61 (d, J = 15.6 Hz, 1H), 1.79-1.70 (m, 3H), 1.66-1.57 (m, 1H), 1.49 (d, J = 12.5 Hz, 1H), 1.07 (d, J = 13.3 Hz, 1H). Its XRPD pattern is basically as shown in Figure 4, indicating that the obtained compound of formula (I) is amorphous.

[0140] Example 1. Preparation of Form I of the Compound of Formula (I)

[0141] About 15 mg of the free base (prepared according to Preparation Example 1-2) was weighed into a 2 mL glass vial, and then about 1 mL of methyl tert-butyl ether solvent was added. After stirring at room temperature for 3 days, the mixture was centrifuged and dried under vacuum.

[0142] The obtained solid was subjected to XRPD detection, and its XRPD pattern is shown in FIG2 , which is defined as the crystalline form I of the compound of formula (I) in the present application. The XRPD pattern of the obtained solid has peaks at the 2θ (°) values ​​shown in the aforementioned Table 1, and the relative intensities of each peak are shown in the aforementioned Table 1; its TGA pattern is shown in FIG3 , and in the TGA curve, the sample loses 0.559% of its weight when heated to about 120° C., so the crystalline form I is an anhydrous substance.

[0143] Example 2. Preparation of Form I of the Compound of Formula (I)

[0144] Referring to the preparation method of Example 1, the methyl tert-butyl ether solvent was replaced with the solvents shown in the following table, and the obtained solid was subjected to XRPD detection. The results are shown in Table 5:

[0145] Table 5

[0146] Example 3. Preparation of Form I of the Compound of Formula (I)

[0147] Weigh about 15 mg of free base (prepared with reference to Preparation Example 1-2) into a 5 mL glass vial, then add 1 mL of solvent. Stir at 60°C for 3 hours, then cool to room temperature and stir overnight, collect wet solid by centrifugation, dry under reduced pressure, and then measure the XRPD of the obtained solid. The solvent and measurement results are shown in Table 6:

[0148] Table 6

[0149] Example 4. Preparation of Form I of the Compound of Formula (I)

[0150] 11.80 g of free base (prepared as described in Preparation Example 1-2) and 120 mL of acetonitrile were added to a 250 mL glass bottle, stirred at room temperature for 8 hours, filtered and dried, and the filter cake was dried under reduced pressure to obtain a solid. The XRPD of the obtained solid was measured, which showed Form I.

[0151] Test Example 1. Thermal stability of Form I

[0152] (1) About 15 mg of Form I (prepared as described in Example 1) was weighed into a 2 mL glass vial, and then about 1 mL of solvent was added. The mixture was stirred overnight at room temperature, centrifuged, and vacuum dried. The XRPD of the solid was then measured. The measurement results of the solvent and the obtained solid are shown in Table 7:

[0153] Table 7

[0154] The results in Table 7 show that Form I can stably exist in a variety of solvents at room temperature.

[0155] (2) Weigh about 15 mg of Form I (prepared with reference to Example 1) into a 5 mL glass vial, then add 1 mL of solvent, stir overnight at 50° C., collect the wet solid by centrifugation, dry under reduced pressure, and then measure the XRPD of the dried solid. The measurement results of the solvent and the obtained solid are shown in Table 8:

[0156] Table 8

[0157] The results in Table 8 show that Form I can stably exist in a variety of solvents under high temperature conditions.

[0158] Test Example 2. Chemical stability test of Form I

[0159] Weigh an appropriate amount of Form I, place it in a weighing bottle, and place it in an open container under high temperature test (60°C) and high humidity test (RH92.5%±5%) conditions for 30 days. Take samples for testing on the 5th, 10th, and 30th days. 6 lux﹒hr, near-ultraviolet lamp energy not less than: 200W﹒hr / m 2 ) for 30 days, and samples were taken for testing on the 5th, 10th and 30th days. The total impurity content was tested by HPLC, and the test results are shown in Table 8-1.

[0160] Table 8-1 Test results of properties and total impurity content

[0161] The results in Table 8-1 show that Form I has high chemical stability.

[0162] Test Example 3. Hygroscopicity test of Form I

[0163] According to the guidelines for the hygroscopicity test of drugs in Part IV 9103 of the 2020 edition of the Chinese Pharmacopoeia, the specific experimental steps are as follows:

[0164] 1. Take two clean, dry, empty weighing bottles and place them in a desiccator (with saturated ammonium chloride solution at the bottom) for 24 hours.

[0165] 2. Take out and weigh accurately after 24 hours (m1), spread an appropriate amount of the test sample in the weighing bottle with a thickness of about 1 mm, weigh accurately (m2), open the bottle mouth, and place it in a desiccator together with the bottle cap for 24 hours.

[0166] 3. Take out the bottle after 24 hours, cover the bottle cap, and weigh it accurately (m3). The experimental results are shown in Table 8-2.

[0167] Table 8-2 Moisture absorption test results

[0168] After being left open for 24 hours, the weight gain percentage of Form I is less than 2% but not less than 0.2%. According to the description of hygroscopic characteristics and the definition of weight gain due to moisture in the Chinese Pharmacopoeia, Form I is slightly hygroscopic.

[0169] Test Example 4. Solubility test of Form I

[0170] The experiment was carried out according to the general rules of Part IV of the 2020 edition of the Chinese Pharmacopoeia, and the results are shown in Table 8-3.

[0171] Table 8-3 Solubility test results

[0172] The results showed that Form I was easily soluble in dimethyl sulfoxide, dissolved in 0.1 mol / L hydrochloric acid solution, and almost insoluble or insoluble in ethanol, ethyl acetate and water.

[0173] Biological Testing

[0174] Test Example 1: Phosphatase activity detection

[0175] IC 50The value was detected by using the Homogeneous Full Length SHP-2 Assay Kit (BPS Bioscience #79330), which uses 6,8-difluoro-4-methylumbelliferyl phosphate (DiFMUP) as the reaction substrate. SHP2 enzyme solution (diluted to 0.04 ng / μl with 1× buffer) was mixed with 0.5 μM SHP-2 activating peptide in 1× buffer solution containing 2.5 mM dithiothreitol (DTT) to activate PTP, and DMSO (0.1% (V / V)), free base of the compound of formula (I) or compound D1 (concentration: 0.5 nM-1 μM) were added and mixed and incubated for 1 hour. DiFMUP (50 μM, total reaction volume of 20 μL) was added to start the reaction. After incubation for 30 min, the fluorescence intensity of the reaction solution was detected using a Tecan Infinite M1000 multifunctional microplate reader (i.e., signal value, excitation light: 350 nm, emission light 460 nm). IC was calculated by XLfit 5.0 software 50 , the test results are shown in Table 9.

[0176] Table 9: IC of compounds of formula (I) against SHP-2 enzyme activity 50 value

[0177] The structure of compound D1 is shown below, and its CAS number is 2377352-41-3.

[0178] As shown in Table 9, the compound of formula (I) of the present invention has a high inhibitory activity against SHP-2 enzyme.

[0179] Test Example 2: Cell Proliferation Detection

[0180] MV-4-11 cells (ATCC#CRL-9591, 20,000 cells / well) were inoculated in a 96-well plate (Corning#3916) and cultured in 80 μL / well of culture medium (culture medium (IMDM) containing 10% fetal bovine serum (FBS), Gibco#10099-141C). After culturing for 24 hours, 20 μL of the free base of the compound of formula (I) or compound D1 configured in different concentrations according to the present invention (DMSO final concentration of 0.1% (V / V)) was added. On the 4th day, 60 μL of Cell Titer-Glo (Promega#G7558) was added to each well at 80 rpm, 25°C, 15 min, and the fluorescence value (i.e., signal value) was detected using a Victor X5 (PE) microplate reader. IC was calculated by XLfit 5.0 software 50 , the test results are shown in Table 10.

[0181] Table 10: IC values ​​of the compounds of formula (I) for the inhibition of MV-4-11 cell proliferation 50 value

[0182] As shown in Table 10, the compound of formula (I) of the present invention has a high inhibitory activity on the proliferation of MV-4-11 cells.

[0183] Test Example 3: Inhibition of hERG potassium channel

[0184] The effect of the free base of the compound of formula (I) on the hERG potassium channel (human Ether-a-go-go Related Gene potassium channel) current was tested using the electrophysiological manual patch clamp method.

[0185] 1 Cell culture and treatment

[0186] CHO cells stably expressing hERG were cultured in a cell culture dish with a diameter of 35 mm, placed in an incubator at 37°C and 5% CO2, and subcultured every 48 hours at a ratio of 1:5. The culture medium formula was: 90% F12 culture medium (Invitrogen), 10% fetal bovine serum (Gibco), 100 μg / mL G418 (Invitrogen) and 100 μg / mL Hygromycin B antibiotic (Invitrogen). On the day of the experiment, the cell culture medium was aspirated, and after rinsing with extracellular fluid, 0.25% trypsin-EDTA (Trypsin-EDTA, Invitrogen) solution was added and digested at room temperature for 3-5 minutes. The digestion solution was aspirated, and the cells were resuspended with extracellular fluid and transferred to a laboratory dish for electrophysiological recording for use.

[0187] 2. Compound Preparation

[0188] On the day of the test, the compound was prepared with DMSO to a 20 mM stock concentration, and then serially diluted 3-fold with DMSO, i.e., 10 μL was added to 20 μL of DMSO, and then 10 μL of the serially diluted compound DMSO solution was added to 4990 μL of extracellular fluid, and the final concentration to be tested was obtained by 500-fold dilution.

[0189] 3 Electrophysiological recording process

[0190] CHO (Chinese Hamster Ovary) cells stably expressing hERG potassium channel were used to record hERG potassium channel current using whole-cell patch clamp technique at room temperature. Glass microelectrodes were pulled from glass electrode blanks (BF150-86-10, Sutter) by a puller. The tip resistance after perfusion of electrode liquid was about 2-5MΩ. The glass microelectrode was inserted into the amplifier probe to connect to the patch clamp amplifier. The clamping voltage and data recording were controlled and recorded by pClamp 10 software through a computer, with a sampling frequency of 10kHz and a filter frequency of 2kHz. After obtaining whole-cell recording, the cell was clamped at -80mV, and the step voltage to induce hERG potassium current (I hERG) was given a 2s depolarization voltage from -80mV to +20mV, then repolarized to -50mV, and returned to -80mV after 1s. This voltage stimulation was given every 10s, and the drug administration process was started after the hERG potassium current was confirmed to be stable (1 minute). Compounds were administered for at least 1 min at each tested concentration, and at least 2 cells (n≥2) were tested at each concentration.

[0191] 4 Data processing

[0192] Data analysis was performed using pClamp 10, GraphPad Prism 5 and Excel software. The degree of inhibition of hERG potassium current (peak value of hERG tail current induced at -50 mV) by different compound concentrations was calculated using the following formula: Inhibition% = [1–(I / Io)] × 100%. Inhibition% represents the percentage of inhibition of hERG potassium current by the compound, and I and Io represent the amplitude of hERG potassium current after and before drug addition, respectively.

[0193] Compound IC 50 The calculation was performed using GraphPad Prism 5 software by fitting the following equation: Y = Bottom + (Top-Bottom) / (1 + 10^((LogIC 50 -X)×HillSlope)). Wherein, X is the Log value of the compound detection concentration, Y is the inhibition percentage at the corresponding concentration, Bottom and Top are the minimum and maximum inhibition percentages measured at each compound concentration during the test, respectively; HillSlope is the slope factor. The results of the inhibition of hERG potassium channel current by the compound of formula (I) are shown in Table 11.

[0194] Table 11: IC of hERG potassium channel current inhibition by compounds of formula (I) 50 value

[0195] It can be seen from the data in Table 11 that the compound of formula (I) of the present invention has a weaker inhibitory effect on hERG current and has good safety.

[0196] Test Example 4: In vivo efficacy experiment

[0197] 1.KYSE-520 subcutaneous transplant tumor model

[0198] The culture conditions of human esophageal squamous cell carcinoma cell line KYSE-520 are RPMI 1640 medium containing 10% fetal bovine serum, placed in a 37℃ 5% CO2 sterile incubator. Subculture 2-3 times a week, digest with 0.25% trypsin-EDTA, terminate digestion with the above complete medium, centrifuge at 1000rpm for 5 minutes, discard the supernatant, resuspend the cells and subculture at a ratio of 1:3-1:5. Collect cells in the logarithmic growth phase, resuspend the cells in RPMI 1640 medium without serum, count and adjust to the appropriate density.

[0199] KYSE-520 cells (3×10 6 The tumors were inoculated into the right ventral back of female Balb / C nude mice of about 6-8 weeks old. When the tumors grew to an average volume of about 200 mm 3 Around 14:00, the patients were randomly divided into groups according to tumor volume and body weight (n=8-10). The drug was administered once a day starting on the day of grouping, and the day of grouping was defined as day 0. The tumor diameter was measured with a vernier caliper twice a week, and the tumor volume was calculated as TV (mm 3 )=0.5×a×b 2 , a and b represent the long and short diameters of the tumor, respectively (in mm). When the average tumor volume of the vehicle control group was about 800 mm 3 The experiment was terminated at 1:00 pm, the tumor tissue was separated and weighed, and the tumor weight proliferation rate T / C was calculated. 重量 % = TW T / TW C %,TW T and TW C The daily dosage and the obtained tumor weight proliferation rate are shown in Table 12.

[0200] Table 12: Tumor proliferation rate of the compound of formula (I) of the present invention

[0201] 2. MC38 subcutaneous tumor model

[0202] The culture conditions of mouse colon cancer cell line MC38 are RPMI 1640 medium containing 10% fetal bovine serum, placed in a 37℃ 5% CO2 sterile incubator. Subculture 2-3 times a week, digest with 0.25% trypsin-EDTA, terminate digestion with the above complete medium, centrifuge at 1000rpm for 5 minutes, discard the supernatant, resuspend the cells and subculture at a ratio of 1:4-1:8. Collect cells in the logarithmic growth phase, resuspend the cells in RPMI 1640 medium without serum, count and adjust to the appropriate density.

[0203] MC38 cells (5×10 6 The tumors were inoculated into the right ventral back of female C57BL / 6 mice of about 6-8 weeks old when they grew to an average volume of about 50-100 mm. 3 Around 14:00, the patients were randomly divided into groups according to tumor volume and body weight (n=8-10). The patients were orally administered 10 mg / kg once a day starting from the day of grouping, and the day of grouping was defined as day 0. The tumor diameter was measured with a vernier caliper twice a week, and the tumor volume was calculated as TV (mm 3 )=0.5×a×b 2 , a and b represent the long and short diameters of the tumor, respectively (in mm). When the average tumor volume of the vehicle control group was about 1800 mm 3 The experiment was terminated at 1:00 pm, the tumor tissue was separated and weighed, and the tumor weight proliferation rate T / C was calculated. 重量 % = TW T / TW C %,TW T and TW C The test results are shown in Table 13.

[0204] Table 13: Tumor proliferation rate of the compound of formula (I) of the present invention

[0205] The results in Table 12 and Table 13 show the tumor growth rate T / C of the compound of formula (I) in the KYSE-520 subcutaneous transplant tumor model and the MC38 subcutaneous tumor model. 重量 % were all less than 30%, showing a significant effect in inhibiting tumor growth and having good anti-tumor efficacy.

[0206] All documents mentioned in the present invention are cited as references in this application, just as each document is cited as reference separately. In addition, it should be understood that after reading the above content of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the claims attached to this application.

[0207] The technical features of the above-described embodiments may be arbitrarily combined. To make the description concise, not all possible combinations of the technical features in the above-described embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0208] The above-mentioned embodiments only express several implementation methods of the present invention, and the description thereof is relatively specific and detailed, but it cannot be understood as limiting the scope of the invention patent. It should be pointed out that for ordinary technicians in this field, several variations and improvements can be made without departing from the concept of the present invention, which all belong to the protection scope of the present invention. Therefore, the protection scope of the patent of the present invention shall be based on the attached claims, and the description and drawings can be used to interpret the content of the claims.

[0209] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

A solid form of a compound of formula (I), The solid forms are polymorphs. The solid form according to claim 1, characterized in that The polymorph is Form I, and its XRPD pattern 2θ (°) diffraction angle has characteristic diffraction peaks at 15.982±0.2, 20.367±0.2 and 21.114±0.

2. The solid form according to claim 1, characterized in that The polymorph is Form I, and its XRPD pattern 2θ (°) diffraction angle has characteristic diffraction peaks at 8±0.2, 9.232±0.2, 11.088±0.2, 12.438±0.2, 13.315±0.2, 13.586±0.2, 14.598±0.2, 15.025±0.2, 15.982±0.2, 17.181±0.2, 19.523±0.2, 19.943±0.2, 20.367±0.2 and 21.114±0.

2. The solid form according to claim 1, characterized in that The polymorph is Form I, and its XRPD pattern is substantially as shown in FIG. 2 . The solid form according to any one of claims 1 to 4, characterized in that The polymorph is Form I and has a TGA pattern substantially as shown in FIG. 3 . A method for preparing the solid form according to claim 1, characterized in that: The following steps are involved: a. dispersing the free base of the compound of formula (I) in a solvent to obtain a liquid; b. The liquid in step (a) is subjected to crystallization to obtain a solid form of the compound of formula (I). The preparation method according to claim 6, characterized in that The free base of the compound of formula (I) is amorphous. A pharmaceutical composition, comprising: A solid form of a compound of formula (I) according to any one of claims 1 to 5, and a pharmaceutically acceptable carrier. Use of a solid form of a compound of formula (I) according to any one of claims 1 to 5, or a pharmaceutical composition according to claim 8, in the preparation of a medicament for treating and / or preventing a disease or condition mediated by SHP2 or associated with abnormal SHP2 activity. A method for treating a disease or condition mediated by SHP2 or associated with abnormal SHP2 activity, the method comprising administering to a subject a therapeutically effective amount of a solid form of a compound of formula (I) according to any one of claims 1 to 5, or a pharmaceutical composition according to claim 8.

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