Pharmaceutical salts of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1h-pyrazol-3-yl]amino]pyrazine-2-carbonitrile, pharmaceutical composition and combination comprising them, use thereof, and method for their preparation

Abstract

This invention provides a pharmaceutically acceptable salt of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyr-2-carboxynitrile, selected from maleic acid salts, toluenesulfonates, benzenesulfonates, and malonates. Specific crystalline forms of these salts, methods for preparing these salts, pharmaceutical compositions containing these salts, and their therapeutic uses are also provided.

Description

Technical Field

[0001] This invention relates to a pharmaceutical salt of the Chk-1 inhibitor compound 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxynitrile, a method for its preparation, pharmaceutical compositions containing the same, and its use in the treatment of diseases such as cancer. Prior Technology

[0002] Chk-1 is a serine / threonine kinase involved in inducing cell cycle checkpoints in response to DNA damage and replication stress [Tse ​​et al., Clin. Can. Res. 2007;13(7)]. Cell cycle checkpoints are regulatory pathways that control the sequence and timing of cell cycle transitions. G1 checkpoint activation is impaired in many cancer cells. For example, Hahn et al. and Hollstein et al. have reported that tumors are associated with mutations in the p53 gene, a tumor suppressor gene found in approximately 50% of all human cancers [N Engl J Med 2002, 347(20):1593; Science, 1991, 253(5015):49].

[0003] Chk-1 inhibition eliminates checkpoints within S and G2 / M and has been shown to selectively sensitize tumor cells to well-known DNA damaging agents. Examples of DNA-damaging agents that have demonstrated this sensitizing effect include gemcitabine, pemetrexed, cytarabine, irinotecan, camptothecin, cisplatin, carboplatin [Clin. Cancer Res. 2010, 16, 376], temozolomide [Journal of Neurosurgery 2004, 100, 1060], doxorubicin [Bioorg. Med. Chem. Lett. 2006;16:421-6], paclitaxel [WO2010149394], and hydroxyurea [Nat. Cell. Biol.]. [2005;7(2):195-20], nitroimidazole hypoxia-targeting drug TH-302 (Meng et al., AACR, 2013 Abstract No. 2389) and ionizing radiation [Clin. Cancer Res. 2010, 16, 2076]. See also the commentary by McNeely et al., [Pharmacology & Therapeutics (2014), 142(1):1-10].

[0004] Recent data also show that Chk-1 inhibitors can work synergistically with the following: PARP inhibitors [Cancer Res 2006.; 66:(16)], Mek inhibitors [Blood. 2008; 112(6): 2439-2449], Farnesyltransferase inhibitors [Blood. 2005;105(4):1706-16], Rapamycin [Mol. Cancer Ther. 2005;4(3):457-70], Src inhibitors [Blood. 2011;117(6):1947-57], and WEE1 inhibitors [Carrassa, 2021, 11(13):2507; Chaudhuri et al., Haematologica, 2014 99(4):688.].

[0005] Furthermore, Chk-1 inhibitors have shown advantages when combined with immunotherapies [Mouw et al., Br J Cancer, 2018. (7):933]. Chk1 inhibitors have been shown to activate cGAS, which induces innate immune responses via STING signaling, and induces PD-L1 expression in vivo and synergizes with anti-PD-L1 [Sen et al., Cancer Discov 2019 (5):646; Sen et al., J Thorac Oncol, 2019. (12):2152].

[0006] A clinical problem of resistance to chemotherapy and radiotherapy is associated with the activation of the DNA damage response involved by Chk-1 [Nature; 2006; 444(7):756-760;Biochem. Biophys. Res. Commun.2011;406(1):53-8].

[0007] It is also envisioned that Chk-1 inhibitors, either as single agents or in combination, could be used to treat tumor cells in which constitutive activation of DNA damage and checkpoint pathways drive genomic instability, particularly through replication pressure. This phenotype is associated with complex karyotypes, such as in samples from patients with acute myeloid leukemia (AML) [Cancer Research 2009, ].

[89] , 8652]. In vitro antagonism of Chk-1 kinase by small molecule inhibitors or by RNA interference significantly reduced the adult characteristics of AML samples with high DNA damage levels. In contrast, Chk-1 inhibition had no effect on normal hematopoietic precursor cells. In addition, recent studies have shown that the tumor microenvironment drives genetic instability [Nature; 2008;(8):180-192] and that loss of Chk-1 makes cells sensitive to hypoxia / reoxygenation [Cell Cycle; 2010; 9(13):2502]. In neuroblastoma, kinase somatic RNA interference screening showed that loss of Chk-1 inhibited the growth of eight neuroblastoma cell lines. Fanconi anemia tumor cells with defective DNA repair showed sensitivity to Chk-1 inhibition [Molecular Cancer 2009, 8652]. [8]:24]. It has been shown that the Chk-1 specific inhibitor PF-00477736 inhibits 30 ovarian cancer cell lines [Bukczynska et al., 23rd Lorne Cancer Conference] and triple-negative breast cancer cells [Cancer Science 2011,

[0102] The growth of , 882]. In addition, PF-00477736 showed selective single-agent activity in a mouse model of spontaneous cancer driven by the MYC oncogene [Ferrao et al., Oncogene (August 15, 2011)]. Chk-1 was inhibited by RNA interference or selective small molecule inhibitors to induce apoptosis in mice with overexpressing MYC in vitro and in vivo B-cell lymphoma models [Höglund et al., Clinical Cancer Research, 2011]. The following data indicate that Chk-1 inhibitors can be used to treat MYC-driven malignancies such as B-cell lymphoma / leukemia, neuroblastoma, and some breast and lung cancers. Chk-1 inhibitors have also shown efficacy in pediatric tumor models, including Ewing's sarcoma and rhabdomyosarcoma [Lowery, 2018. Clin Cancer Res 2019, 25(7):2278]. Chk1 inhibitors have been shown to be synthetically lethal with the DNA polymerase B family, leading to increased replication stress, DNA damage, and cell death [Rogers et al., 2020, 80(8);1735]. Other cell cycle regulatory genes, including CDK2 and POXM1, have also been reported to confer sensitivity to Chk-1 inhibitors [Ditano et al., 2020, 1.11(1);7077;Branigan et al., 2021, Cell Reports 34(9):1098808].

[0008] Mutations that reduce DNA repair pathway activity have also been reported to lead to synthetic lethal interactions with Chk1 inhibition. For example, mutations disrupting the RAD50 complex and ATM signaling increase responsiveness to Chk1 inhibition [Al-Ahmadie et al., Cancer Discov. 2014. (9):1014-21]. Similarly, defects in the homologous DNA repair pathway in Fanconi anemia lead to sensitivity to Chk1 inhibition [Chen et al., Mol. Cancer 2009 8:24, Duan et al., Frontiers in Oncology 2014 4:368]. Furthermore, human cells with loss of function in the Rad17 gene product are sensitive to Chk1 inhibition [Shen et al., Oncotarget, 2015. 6(34):35755].

[0009] Various attempts have been made to develop inhibitors of the Chk-1 kinase. For example, WO 03 / 10444 and WO 2005 / 072733 (both under the name Millennium) disclose aryl / heteroaryl urea compounds as Chk-1 kinase inhibitors. US2005 / 215556 (Abbott) discloses macrocyclic urea as a kinase inhibitor. WO 02 / 070494, WO2006014359, and WO2006021002 (all under the name Icos) disclose aryl and heteroaryl urea as Chk-1 inhibitors. WO / 2011 / 141716 and WO / 2013 / 072502 both disclose substituted pyridyl-phenylurea as Chk-1 kinase inhibitors. WO2005 / 009435 (Pfizer) and WO2010 / 077758 (Eli Lilly) reveal aminopyrazole as a Chk-1 kinase inhibitor.

[0010] WO2015 / 120390 discloses a class of substituted phenyl-pyrazolylamines as Chk-1 kinase inhibitors. One of the disclosed compounds is 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxynitrile, the synthesis of which is described in Example 64 and Synthetic Method L of WO2015 / 12039. This compound is disclosed in its hydrochloride form.

[0011] WO2018 / 183891 (Cascadian Therapeutics) discloses the compound 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazol-2-carboxylonitrile or a pharmaceutically acceptable salt thereof in combination with a WEE-1 inhibitor. However, specific salts of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazol-2-carboxylonitrile are not disclosed. Invention content

[0012] It has been discovered that 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxynitrile forms crystalline salts with various inorganic and organic acids.

[0013] Therefore, in the first embodiment (Example 1.1), the present invention provides a pharmaceutically acceptable salt of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxynitrile, selected from hydrobromide, methanesulfonate, L-tartaric acid Salts, ethanesulfonates, L-aspartate salts, benzenesulfonates, toluenesulfonates, sulfates, phosphates, citrates, acetates, L-glutamates, cis-butenedioic acid salts, cholates, glucuronates, malonates, naphthyl-2-sulfonates, ethane-1,2-disulfonates, naphthalene-1,5-disulfonates, and oxalates.

[0014] The terms "hydrobromate, methanesulfonate, L-tartrate, ethanesulfonate, L-aspartate, benzenesulfonate, toluenesulfonate, sulfate, phosphate, citrate, acetate, L-glutamate, maleate, gentianate, glucuronate, malonate, naphthyl-2-sulfonate, and oxalate" are used in this article for their common meanings. Used to indicate salts formed from hydrobromic acid, methanesulfonic acid, L-tartaric acid, ethanesulfonic acid, L-aspartic acid, benzenesulfonic acid, p-toluenesulfonic acid, sulfuric acid, phosphoric acid, citric acid, acetic acid, L-glutamic acid, maleic acid, gentian acid, glucuronic acid, malonic acid, naphthyl-2-sulfonic acid, ethane-1,2-disulfonic acid, naphthalene-1,5-disulfonic acid, and oxalic acid, respectively.

[0015] Several salts of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxynitrile have also been found to have improved properties compared to the hydrochlorides disclosed in WO2015 / 12039.

[0016] Therefore, in another embodiment (Example 1.2), the present invention provides a pharmaceutically acceptable salt of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxynitrile, which is selected from maleate, toluenesulfonate, benzenesulfonate and malonate.

[0017] The compound 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazol-2-carboxylonite has the following formula (1), and the maleic acid salt, toluene sulfonate, benzene sulfonate and malonate of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazol-2-carboxylonite may be referred herein as salts of the compound of formula (1) or salts of the present invention for convenience.

[0018] Compounds of formula (1) have a number of basic nitrogen atoms and, in principle, can form salts with different salt ratios (i.e., the molar ratio of free base to acid). For example, when the acid is a monocarboxylic acid, a single salt (i.e., where the molar ratio of acid to free base is 1:1) or a double salt (where the molar ratio of acid to free base is approximately 2:1) can be prepared according to the molar equivalent number of the acid used in the method for salt formation. When a dicarboxylic acid (e.g., a dicarboxylic acid) is used for salt formation, half salts (where the molar ratio of acid to base in the salt is 0.5:1), single salts, and double salts can be formed depending on the specific salt formation conditions used.

[0019] Therefore, in other embodiments (Examples 1.3 to 1.9), the present invention provides: 1.3 A pharmaceutically acceptable salt of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxynitrile, which is a maleate. 1.3A A pharmaceutically acceptable salt of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxynitrile, which is a crystalline maleate having crystal pattern B as defined herein. 1.4 A pharmaceutically acceptable salt of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxynitrile, which is a toluenesulfonate salt. 1.5 A pharmaceutically acceptable salt of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxynitrile, which is a benzenesulfonate salt. 1.6 A pharmaceutically acceptable salt of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxynitrile, which is a malonate. 1.7 A pharmaceutically acceptable salt, such as any of Examples 1.1 to 1.6, having a salt ratio of approximately 1:1 (molar ratio of acid to free base). 1.8 A pharmaceutically acceptable salt, such as that of Example 1.3 or Example 1.6, having a salt ratio of approximately 0.5:1 (molar ratio of acid to free base). 1.9 A pharmaceutically acceptable salt, such as that of Example 1.3 or Example 1.5, having a salt ratio of approximately 2:1 (molar ratio of acid to free base).

[0020] The salts of the compounds in formula (1) can be amorphous or substantially crystalline.

[0021] The term "substantially crystalline" refers to salt that is 50% to 100% crystalline. Within this range, the salt may be at least 55% crystalline, or at least 60% crystalline, or at least 70% crystalline, or at least 80% crystalline, or at least 90% crystalline, or at least 95% crystalline, or at least 98% crystalline, or at least 99% crystalline, or at least 99.5% crystalline, or at least 99.9% crystalline, such as 100% crystalline.

[0022] Some of the salts of this invention can exist in several different crystalline or polymorphic forms.

[0023] In UK Patent Application No. 2107924.9, filed June 3, 2021, which claims priority in this application, certain forms of maleic anhydride were designated as Figure A, Figure A', and Figure A''. These forms have been redesignated in this application as Figure A, Figure B, and Figure C, respectively.

[0024] The salt of the compound of formula (1) is preferably crystallized with a purity of at least 90%, more preferably at least 95%; that is, at least 90% (more preferably at least 95%) of the salt has a single crystallized form.

[0025] The salt of the present invention may be in the form of a solvated (e.g., hydrated) or a non-solvated (e.g., anhydrous) crystal.

[0026] As used herein, the term "anhydrous" does not preclude the possibility that some water may be present on or within the crystalline form of the salt. For example, some water may be present on the surface of the crystalline form of the salt, or a small amount of water may be present within the body of the crystalline form of the salt. Typically, the anhydrous form contains less than 0.4 water molecules per molecule of formula (1), and more preferably less than 0.1 water molecules per molecule of formula (1), for example, 0 water molecules.

[0027] When hydrated in its crystalline form, it may contain, for example, up to three water molecules of crystallization, and more typically up to two water molecules, such as one or two water molecules. It may also form non-stoichiometric hydrates, in which the number of water molecules present is less than one, or otherwise a non-integer number. For example, when less than one water molecule is present, each molecule of compound (1) may contain, for example, 0.4, 0.5, 0.6, 0.7, 0.8, or 0.9 water molecules.

[0028] Therefore, in other embodiments of the present invention (Examples 1.10 to 1.11), the following is provided: 1.10 A pharmaceutically acceptable salt as described in any of Examples 1.1 to 1.9, which is 50% to 100% crystalline. 1.11 The pharmaceutically acceptable salt as described in Example 1.10 is: (b) At least 55% crystallization; or 1.11A A pharmaceutically acceptable salt as described in Example 1.11, which is at least 60% crystallized. 1.11B A pharmaceutically acceptable salt as described in Example 1.11, which is at least 70% crystallized. 1.11C A pharmaceutically acceptable salt as described in Example 1.11, which is at least 80% crystallized. 1.11D A pharmaceutically acceptable salt as described in Example 1.11, which is at least 90% crystallized. 1.11E A pharmaceutically acceptable salt as described in Example 1.11, which is at least 95% crystallized. 1.11F A pharmaceutically acceptable salt as described in Example 1.11, which is at least 98% crystallized. 1.11G A pharmaceutically acceptable salt as described in Example 1.11, which is at least 99% crystallized. 1.11H A pharmaceutically acceptable salt as described in Example 1.11, which is at least 99.5% crystallized. 1.11I The pharmaceutically acceptable salt as described in Example 1.11, which is at least 99.9% crystallized. 1.11J A pharmaceutically acceptable salt as described in Example 1.11, which is 100% crystalline.

[0029] Crystallographic morphology can be characterized using a variety of techniques, including X-ray powder diffraction (XRPD), single-crystal X-ray diffraction, differential scanning calorimetry (DSC), and pyrolysis gravimetric analysis (TGA). The behavior of crystals under different humidity conditions can be analyzed using gravimetric vapor-phase adsorption (GVS) studies, such as dynamic vapor-phase adsorption (DVS).

[0030] The crystalline structure of compounds can be analyzed using solid-state techniques such as X-ray powder diffraction (XRPD). XRPD can be performed using conventional methods, such as those described herein (see examples below) and those in "Introduction to X-ray Powder Diffraction," Ron Jenkins and Robert L. Snyder (John Wiley & Sons, New York, 1996). The presence of defined peaks (opposite to random background noise) in the XRPD diffraction pattern indicates a certain degree of crystallinity in the compound.

[0031] The X-ray powder pattern of the compound is characterized by the diffraction angle (2θ) of the X-ray diffraction spectrum (also referred to as °2Th or °2θ in this paper) and the interplanar spacing (d). These are related to Bragg's equation, nλ = 2d Sin θ, where n = 1; λ = wavelength of X-ray radiation; d = interplanar spacing; and θ = diffraction angle.

[0032] Therefore, in other embodiments (Examples 1.12 to 1.42), the present invention provides: 1.12 A pharmaceutically acceptable salt of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxynitrile as in Example 1.1, having an XRPD spectrum substantially as shown in any of Figures 5 to 25, 27, 29, 31, 33 and 35 (without regard to any XRPD spectrum in the form of a free base or amorphous salt). 1.13 A pharmaceutically acceptable maleate pattern B salt of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxynitrile, as described in any of Examples 1.2, 1.3 and 1.3A, having an XRPD spectrum substantially as shown in Figure 25. 1.14 A pharmaceutically acceptable maleate pattern B salt of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxylonite as described in Examples 1.2, 1.3, 1.3A, 1.12 and 1.13, having an XRPD spectrum (e.g., with 100% relative intensity) characterized by a dominant °2Th (°2θ) peak at 26.3 ± 0.2º. 1.14A A pharmaceutically acceptable maleate pattern B salt of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxylonite, as described in Examples 1.2, 1.3, 1.3A, 1.12 and 1.13, having an XRPD spectrum characterized by a dominant °2Th (°2θ) peak at 6.9 ± 0.2º and / or 26.4 ± 0.2º and / or 11.8 ± 0.2º and / or 17.9 ± 0.2º. 1.15 A pharmaceutically acceptable maleate patterned B salt as described in Example 1.14, having an XRPD spectrum characterized by a dominant °2Th peak at 6.9 ±0.2º, 26.4 ±0.2º, 11.8 ±0.2º and 17.9 ±0.2º. 1.16 A pharmaceutically acceptable maleate pattern B salt as described in Example 1.14 or Example 1.15, having an XRPD spectrum characterized by an intermediate °2Th peak at 15.6 ±0.2º and / or 9.4 ±0.2º and / or 15.8 ±0.2º and / or 17.7 ±0.2º and / or 26.8 ±0.2º. 1.17 The pharmaceutically acceptable maleate pattern B salt of Example 1.16, having an XRPD spectrum characterized by an intermediate °2Th peak at 15.6 ±0.2º, 9.4 ±0.2º, 15.8 ±0.2º, 17.7 ±0.2º and 26.8 ±0.2º. 1.18 The pharmaceutically acceptable maleate pattern A salt of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxynitrile, as in Example 1.2, has an XRPD spectrum substantially as shown in Figure 27. 1.19 A pharmaceutically acceptable maleate pattern A salt of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxylonite, as described in Example 1.2 or Example 1.18, having an XRPD spectrum characterized by a dominant 2Th peak at 6.6 ± 0.2º and / or 17.3 ± 0.2º and / or 11.1 ± 0.2º. 1.20 A pharmaceutically acceptable maleate pattern A salt of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxylonite, as described in Example 1.19, having a characteristic XRPD spectrum with main 2Th peaks at 6.6 ± 0.2, 17.3 ± 0.2º, and 11.1 ± 0.2º. 1.21 A pharmaceutically acceptable maleate pattern A salt as described in Example 1.19 or Example 1.20, having an XRPD spectrum characterized by an intermediate °2Th peak at 26.5 ±0.2º and / or 9.2 ±0.2º and / or 14.3 ±0.2º and / or 18.5 ±0.2º and / or 25.9 ±0.2º and / or 11.5 ±0.2º and / or 16.9 ±0.2º and / or 20.5 ±0.2º and / or 15.6 ±0.2º. 1.22 The pharmaceutically acceptable maleate pattern A salt as described in Example 1.21, having an XRPD spectrum characterized by an intermediate °2Th peak at 26.5 ±0.2º, 9.2 ±0.2º, 14.3 ±0.2º, 18.5 ±0.2º, 25.9 ±0.2º, 11.5 ±0.2º, 16.9 ±0.2º, 20.5 ±0.2º and 15.6 ±0.2º. 1.23 The pharmaceutically acceptable maleic acid salt patterned C salt of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxynitrile, as in Example 1.2, has an XRPD spectrum substantially as shown in Figure 29. 1.24 A pharmaceutically acceptable maleic anhydride patterned C salt of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxylonite, as described in Example 1.2 or Example 1.23, having an XRPD spectrum characterized by a dominant 2Th peak at 6.7 ± 0.2º and / or 9.2 ± 0.2º and / or 11.5 ± 0.2º and / or 15.6 ± 0.2º and / or 17.4 ± 0.2º and / or 17.7 ± 0.2º and / or 26.3 ± 0.2º. 1.25 A pharmaceutically acceptable maleic acid ester patterned C salt of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxylonite, as described in Example 1.24, having an XRPD spectrum characterized by a dominant 2Th peak at 6.7 ±0.2º, 9.2 ±0.2º, 11.5 ±0.2º, 15.6 ±0.2º, 17.4 ±0.2º, 17.7 ±0.2º and 26.3 ±0.2º. 1.26 A pharmaceutically acceptable maleate patterned C salt as described in Example 1.24 or Example 1.25, having an XRPD spectrum characterized by an intermediate °2Th peak at 18.5 ±0.2º and / or 14.3 ±0.2º and / or 21.7 ±0.2º and / or 11.1 ±0.2º and / or 27.6 ±0.2º and / or 17.0 ±0.2º and / or 25.6 ±0.2º and / or 16.0 ±0.2º and / or 22.2 ±0.2º. 1.27 A pharmaceutically acceptable maleate patterned C salt as described in Example 1.26, having an XRPD spectrum characterized by an intermediate °2Th peak at 18.5 ±0.2º, 14.3 ±0.2º, 21.7 ±0.2º, 11.1 ±0.2º, 27.6 ±0.2º, 17.0 ±0.2º, 25.6 ±0.2º, 16.0 ±0.2º, and 22.2 ±0.2º. 1.28 The pharmaceutically acceptable malonate pattern B salt of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxynitrile, as in Example 1.12, has an XRPD spectrum substantially as shown in Figure 31. 1.29 A pharmaceutically acceptable malonate pattern B salt of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxynitrile, as in Example 1.2 or Example 1.28, having an XRPD spectrum characterized by a dominant 2Th peak at 10.6 ± 0.2º and / or 6.5 ± 0.2º. 1.30 A pharmaceutically acceptable malonate pattern B salt of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxynitrile, as described in Example 1.29, having an XRPD spectrum characterized by dominant 2Th peaks at 10.6 ± 0.2º and 6.5 ± 0.2º. 1.31 A pharmaceutically acceptable malonate pattern B salt of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxynitrile, as described in Example 1.29 or Example 1.30, having an XRPD spectrum characterized by an intermediate °2Th peak at 16.6 ±0.2º and / or 18.4 ±0.2º and / or 14.3 ±0.2º and / or 25.9 ±0.2º. 1.32 The pharmaceutically acceptable malonate pattern B salt of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxynitrile, as described in Example 1.31, has an XRPD spectrum characterized by an intermediate °2Th peak at 16.6 ±0.2º, 18.4 ±0.2º, 14.3 ±0.2º and 25.9 ±0.2º. 1.33 The pharmaceutically acceptable toluenesulfonate pattern A salt of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxynitrile, as in Example 1.2, has an XRPD spectrum substantially as shown in Figure 33. 1.34 A pharmaceutically acceptable toluenesulfonate pattern A salt of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxynitrile, as described in Example 1.2 or Example 1.33, having an XRPD spectrum characterized by a dominant 2Th peak at 9.1 ± 0.2º and / or 22.2 ± 0.2º and / or 14.9 ± 0.2º and / or 13.8 ± 0.2º. 1.35 A pharmaceutically acceptable toluenesulfonate pattern A salt of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxynitrile, as described in Example 1.34, having an XRPD spectrum characterized by a dominant 2Th peak at 9.1 ± 0.2º, 22.2 ± 0.2º, 14.9 ± 0.2º, and 13.8 ± 0.2º. 1.36 A pharmaceutically acceptable toluenesulfonate pattern A salt of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxynitrile, as described in Example 1.34 or Example 1.35, having an XRPD spectrum characterized by an intermediate °2Th peak at 11.7 ±0.2º and / or 8.8 ±0.2º and / or 15.7 ±0.2º and / or 17.9 ±0.2º and / or 16.5 ±0.2º and / or 24.8 ±0.2º and / or 22.6 ±0.2º. 1.37 A pharmaceutically acceptable toluenesulfonate pattern A salt of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxynitrile, as described in Example 1.36, having an XRPD spectrum characterized by an intermediate °2Th peak at 11.7 ±0.2º, 8.8 ±0.2º, 15.7 ±0.2º, 17.9 ±0.2º, 16.5 ±0.2º, 24.8 ±0.2º, and 22.6 ±0.2º. 1.38 The pharmaceutically acceptable benzenesulfonate pattern C salt of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxynitrile, as in Example 1.2, having an XRPD spectrum substantially as shown in Figure 35. 1.39 A pharmaceutically acceptable benzenesulfonate patterned C salt of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxynitrile, as described in Example 1.2 or Example 1.38, having an XRPD spectrum characterized by a dominant 2Th peak at 15.5 ±0.2º and / or 14.7 ±0.2º and / or 25.4 ±0.2º and / or 20.9 ±0.2º and / or 18.1 ±0.2º and / or 11.2 ±0.2º and / or 13.3 ±0.2º and / or 16.1 ±0.2º. 1.40 A pharmaceutically acceptable benzenesulfonate patterned C salt of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxynitrile, as described in Example 1.39, having an XRPD spectrum characterized by a dominant 2Th peak at 15.5 ±0.2º, 14.7 ±0.2º, 25.4 ±0.2º, 20.9 ±0.2º, 18.1 ±0.2º, 11.2 ±0.2º, 13.3 ±0.2º, and 16.1 ±0.2º. 1.41 A pharmaceutically acceptable benzenesulfonate patterned C salt of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxynitrile, as described in Example 1.39 or Example 1.40, having an XRPD spectrum characterized by an intermediate °2Th peak at 24.1 ±0.2º and / or 9.4 ±0.2º and / or 26.4 ±0.2º and / or 16.3 ±0.2º and / or 19.2 ±0.2º and / or 27.0 ±0.2º. 1.42 A pharmaceutically acceptable benzenesulfonate patterned C salt of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxynitrile, as described in Example 1.41, having an XRPD spectrum characterized by an intermediate °2Th peak at 24.1 ±0.2º, 9.4 ±0.2º, 26.4 ±0.2º, 16.3 ±0.2º, 19.2 ±0.2º and 27.0 ±0.2º.

[0033] In the above embodiments, the term "main 2Th peak" refers to a peak with a relative intensity of at least 50% (relative to the maximum peak), while the term "intermediate peak" refers to a peak with a relative intensity between 20% and 50%. Peak positions are accurate to one decimal place ±0.2º, although they have been measured to at least four decimal places. Peak positions are typically listed in descending order of relative intensity.

[0034] The salts of this invention can also be characterized by their thermal behavior, and especially by their DSC and TGA analyses. Therefore, in other embodiments, the invention provides: 1.43 A pharmaceutically acceptable maleate pattern B salt of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxynitrile as defined in any of Examples 1.13 to 1.17, having substantially the DSC and TGA characteristics shown in FIG26. 1.44 A pharmaceutically acceptable maleate pattern A salt of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxynitrile as defined in any of Examples 1.18 to 1.22, having substantially the DSC and TGA characteristics shown in FIG28. 1.45 A pharmaceutically acceptable maleate pattern C salt of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxynitrile as defined in any of Examples 1.23 to 1.27, having substantially the DSC and TGA characteristics shown in FIG30. 1.46 A pharmaceutically acceptable malonate pattern B salt of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxynitrile as defined in any of Examples 1.28 to 1.32, having substantially the DSC and TGA characteristics shown in FIG32. 1.47 A pharmaceutically acceptable toluenesulfonate pattern A salt of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxynitrile as defined in any of Examples 1.33 to 1.37, having substantially the DSC and TGA characteristics shown in FIG34. 1.48 A pharmaceutically acceptable benzenesulfonate pattern C salt of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxynitrile as defined in any of Examples 1.38 to 1.42, having substantially the DSC and TGA characteristics shown in FIG36.

[0035] Of the various salts of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxynitrile defined and described above and elsewhere in this text, maleate is the preferred salt.

[0036] The advantage of maleate is that it is crystalline, has a thermodynamically favorable stable form (pattern B), and exhibits a low polymorphic tendency.

[0037] Stability studies conducted over two weeks at 25°C / 60% RH and 40°C / 75% RH showed that maleate remained a free-flowing solid, exhibiting no deliquescence or agglomeration and no change in polymorphism, demonstrating good chemical stability. Subsequent data collected over a six-month period supported these preliminary findings.

[0038] When assessing solubility in biologically relevant solvents, maleate showed improved solubility in water and gastric juice compared to free base. [isotope] [ ]

[0039] Salts as defined in any of Examples 1.1 to 1.48 may contain one or more isotopic substitutions, and references to a particular element include all isotopes of that element within their scope. For example, references to hydrogen include 1H, 2H (D), and 3H (T) within their scope. Similarly, references to carbon and oxygen include 12C, 13C, and 14C, and 16O and 18O, respectively.

[0040] The isotope may be radioactive or non-radioactive. In one embodiment of the invention, the salt does not contain a radioactive isotope. Such compounds are preferably used for therapeutic purposes. However, in another embodiment, the salt may contain one or more radioactive isotopes. Salts containing such radioactive isotopes are suitable for diagnostic applications. [Method for preparing the salt of the present invention]

[0041] The pharmaceutically acceptable salt of the present invention can be prepared from the free base of compound 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxynitrile (formula (1)) by the method illustrated in the following examples. Compound (1) can be prepared by the method described in Example 64, Method L of International Patent Application WO 2015 / 20390, as shown in the following reaction flow 1. [Reaction Flow] [1]

[0042] In other embodiments (Examples 2.1 to 2.10), the present invention provides a method for forming a pharmaceutically acceptable salt of a compound of formula (1): 2.1 A method for preparing a pharmaceutically acceptable salt as defined in Example 1.1 or Example 1.2, comprising dispersing 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxynitrile in tetrahydrofuran to form a mixture, heating the mixture to a high temperature in the range of 45°C to 65°C (e.g., 55°C to 65°C, and especially about 60°C), adding the desired amount of acid to the mixture; maintaining the mixture at or near the high temperature for a defined period of time, and cooling the mixture to separate the pharmaceutically acceptable salt. 2.2 The method as described in Example 2.1, wherein the acid is selected from maleic acid, p-toluenesulfonic acid, benzenesulfonic acid and malonic acid. 2.3 A method for preparing a pharmaceutically acceptable salt as defined in Example 1.1 or Example 1.2, comprising dispersing 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxynitrile in a mixture of tetrahydrofuran and acetonitrile (e.g., a 1:1 mixture) to form a mixture, heating the mixture to a high temperature in the range of 45°C to 55°C (e.g., about 50°C), adding the desired amount of acid to the mixture; maintaining the mixture at or near the high temperature for a defined period of time, and cooling the mixture to separate the pharmaceutically acceptable salt. 2.4 The method as described in Example 2.3, wherein the acid is selected from maleic acid, p-toluenesulfonic acid, benzenesulfonic acid and malonic acid. 2.5 A method for preparing a pharmaceutically acceptable salt as defined in Example 1.1 or Example 1.2, comprising dispersing 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxynitrile in a mixture of tetrahydrofuran and water (e.g., wherein the mixture contains 75% to 97% (v / v) tetrahydrofuran and 3% to 25% (v / v) water, and more preferably about 95% (v / v) tetrahydrofuran and about 5 (v / v) water) to form a mixture, heating the mixture to a high temperature in the range of 45°C to 65°C (e.g., about 50°C to 60°C), adding the desired amount of acid to the mixture; maintaining the mixture at or near the high temperature for a defined period of time, and cooling the mixture to separate the pharmaceutically acceptable salt. 2.6 The method of Example 2.5, wherein the acid is selected from maleic acid, p-toluenesulfonic acid, benzenesulfonic acid and malonic acid. 2.7 The method of Example 2.1, wherein the required amount of acid is an excess acid (e.g., up to 1 mole excess). 2.8 The method as described in Example 2.7, wherein the acid is p-toluenesulfonic acid. 2.9 The method of Example 2.5, wherein the required amount of acid is an excess acid (e.g., up to 1 mole excess). 2.10 The method of Example 2.9, wherein the acid is selected from p-toluenesulfonic acid and benzenesulfonic acid. 2.11 The method of any one of Examples 2.1, 2.3 and 2.5, wherein the acid is maleic acid and the resulting pharmaceutically acceptable salt is maleate. 2.12 The method of Example 2.11, wherein the maleate is the maleate pattern A salt. 2.13 The method of Example 2.13 further includes converting pattern A maleate into pattern B maleate by adjusting pattern A salt in an atmosphere with a relative humidity greater than 50% (e.g., 51% to 90% relative humidity, or 51% to 85% relative humidity). 2.14 The method of Example 2.13, wherein pattern A maleate is conditioned in an atmosphere with a relative humidity greater than 60% and a temperature range of 35-45°C. 2.15 The method of Example 2.13 or Example 2.14, wherein pattern A maleate is conditioned in an atmosphere of 70% to 80% relative humidity. 2.16 The method of Example 2.14, wherein pattern A maleate is conditioned in an atmosphere of approximately 75% relative humidity and approximately 40°C.

[0043] The specific set of conditions used to execute the above methods is illustrated in the following examples. [Biological characteristics and therapeutic uses] [ ]

[0044] The compound of formula (1) and its salts are potent inhibitors of Chk-1, and are therefore expected to be beneficial, either alone or in combination with various chemotherapeutic agents, immunotherapeutic agents or radiation, for the treatment of a wide range of proliferative diseases.

[0045] Therefore, in other embodiments (Examples 3.1 to 3.10), the present invention provides: 3.1 A pharmaceutically acceptable salt as defined in any of Examples 1.1 to 1.48, used in a medicine or therapy. 3.2 A pharmaceutically acceptable salt as defined in any of Examples 1.1 to 1.48, which is used as a Chk-1 kinase inhibitor. 3.3 A medically acceptable salt as defined in any of Examples 1.1 to 1.48, used to enhance the therapeutic effect of radiotherapy, chemotherapy, or immunotherapy in the treatment of proliferative diseases such as cancer. 3.4 A medically acceptable salt as defined in any of Examples 1.1 to 1.48, used to treat proliferative diseases such as cancer. 3.5 Use of a pharmaceutically acceptable salt as defined in any of Examples 1.1 to 1.48, for the manufacture of an agent that enhances the therapeutic effects of radiotherapy, chemotherapy, or immunotherapy in the treatment of proliferative diseases such as cancer. 3.6 Use of a pharmaceutically acceptable salt as defined in any of Examples 1.1 to 1.48, for the manufacture of a medicament for the treatment of proliferative diseases such as cancer. 3.7 A method for preventing or treating proliferative diseases such as cancer, the method comprising administering to a patient, in combination with radiotherapy, immunotherapy or chemotherapy, a medically acceptable salt as defined in any one of Examples 1.1 to 1.48. 3.8 A method for preventing or treating proliferative diseases such as cancer, the method comprising administering to a patient a medically acceptable salt as defined in any one of Examples 1.1 to 1.48. 3.9 A pharmaceutically acceptable salt, use, or method of use as defined in any of Examples 3.3 to 3.8, wherein the cancer is selected from cancerous tumors, such as bladder, brain, breast, colon, kidney, epidermis, liver, lung, esophagus, gallbladder, ovary, pancreas, stomach, cervix, thyroid, prostate, gastrointestinal system, or skin cancer; hematopoietic tumors, such as leukemia, B-cell lymphoma, T-cell lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, pilocellular lymphoma, mantle cell lymphoma, or Burkitt's lymphoma. Lymphoma); hematopoietic tumors of the bone marrow spectrum, such as acute and chronic myeloid leukemia, myelodysplastic syndrome, or promyelocytic leukemia; follicular thyroid carcinoma; mesenchymal cell-derived tumors, such as fibrosarcoma or rhabdomyosarcoma; central or peripheral nervous system tumors, such as astrocytoma, neuroblastoma, glioma, neurotubular cell tumor, or schwannoma; melanoma; seminoma; teratoma; osteosarcoma; xeroderma pigmentosum; keratoacanthoma; follicular thyroid carcinoma; Ewing's sarcoma or Kaposi's sarcoma. 3.10 The pharmaceutically acceptable salt, use or method of use as described in Example 3.9, wherein the cancer is selected from breast cancer, colorectal cancer, lung cancer, ovarian cancer, pancreatic cancer, prostate cancer, glioma, Ewing's sarcoma, lymphoma (e.g., mantle cell lymphoma), neuroblastoma and leukemia.

[0046] It is also conceivable that a medically acceptable salt of formula (1) described herein could be used for treatment: (a) Cancers driven by oncogenes, including Myc and CCNE1; (b) Cancers with dysregulation of cell cycle or DNA damage repair pathways, such as cancers with defects in RAD17 (e.g., RAD17-mutant tumors), RAD50, TP53, or ATM (e.g., tumors with defective DNA repair mechanisms or defective cell cycles, such as cancers with mutations (e.g., in p53) leading to loss of the G1 / S DNA damage checkpoint), or Fanconi anemia; and (c) Cancers such as those with high levels of replication pressure under Chk1 or ATR amplification.

[0047] Therefore, in other embodiments (Examples 3.11 to 3.23), the present invention provides: 3.11 A medically acceptable salt, use, or method of use as defined in any of Examples 3.3 to 3.10, wherein the cancer is characterized by a defective DNA repair mechanism or a defective cell cycle or high level of replication stress. 3.12 The pharmaceutically acceptable salt, use or method of use as described in Example 3.11, wherein the cancer is a p53-negative or mutant tumor. 3.13 A medically acceptable salt, use, or method of use as defined in any of Examples 3.3 to 3.10, wherein the cancer is a MYC oncogene-driven cancer. 3.14 A pharmaceutically acceptable salt, use or method of use as described in Example 3.13, wherein the cancer driven by the MYC oncogene is B-cell lymphoma, leukemia, neuroblastoma, neuroduct-derived tumor, breast cancer or lung cancer. [ ] 3.15 A medically acceptable salt as defined in any of Examples 1.1 to 1.48, used in combination with radiotherapy, immunotherapy or chemotherapy to treat patients with p53-negative or mutant tumors (e.g., cancers selected from breast cancer, colorectal cancer, lung cancer, ovarian cancer, pancreatic cancer, prostate cancer, glioma and leukemia). 3.16 The pharmaceutically acceptable salt for use as described in any of Examples 3.3 to 3.15, wherein in addition to administering the pharmaceutically acceptable salt as defined in any of Examples 1.1 to 1.48, the treatment also includes administering to the patient a chemotherapeutic agent selected from cytarabine, etoposide, gemcitabine, cyclophosphamide, Wee1 inhibitors, and SN-38. 3.17 Use of a pharmaceutically acceptable salt as defined in any of Examples 1.1 to 1.48 for the manufacture of an agent for the treatment of a patient suffering from cancer characterized by a defective DNA repair mechanism or a defective cell cycle or high-level replication stress. 3.18 As used in Example 3.17, wherein the cancer is a p53-negative or mutated tumor. 3.19 A method for treating a patient (e.g., a human patient) with cancer characterized by a defective DNA repair mechanism or a defective cell cycle or high-level replication stress, the method comprising administering to the patient a therapeutically effective amount of a medically acceptable salt as defined in any one of Examples 1.1 to 1.48. 3.20 The method as described in Example 3.19, wherein the cancer is a p53-negative or mutated tumor. 3.21 A pharmaceutically acceptable salt, use or method of use as defined in any of Examples 3.3 to 3.10, wherein the cancer is a RAD17-mutant tumor or an ATM-deficient RAD50-mutant tumor. 3.21 A medically acceptable salt as defined in any of Examples 1.1 to 1.48, used to treat Fanconi anemia. 3.22 Use of a pharmaceutically acceptable salt as defined in any of Examples 1.1 to 1.48, for the manufacture of a medicament for the treatment of Fanconi anemia. 3.23 A method for treating Fanconi anemia in an individual (e.g., a human individual), the method comprising administering to the individual a therapeutically effective amount of a medically acceptable salt as defined in any one of Examples 1.1 to 1.48.

[0048] The Chk-1 inhibitor salt of this invention can be used alone or in combination with DNA-damaging anticancer drugs and / or radiation therapy and / or immunotherapy to treat individuals with multidrug-resistant cancer. Cancer is considered drug-resistant when it returns to a normal tumor growth rate after initial drug response and subsequent drug treatment. A tumor is considered "drug-responsive" when it shows a decrease in size or growth rate.

[0049] Before administering a medically acceptable salt as defined in any of Examples 1.1 to 1.48, patients may be screened to determine whether the cancer they have or may have is one that is sensitive to treatment with a combination of a Chk-1 kinase inhibitor compound or a chemotherapy agent (such as a DNA damaging agent) and a Chk-1 kinase inhibitor compound.

[0050] More specifically, patients can be screened to determine whether the cancer they have or may have is a cancer characterized by defective DNA repair mechanisms or defective cell cycle or high levels of replication stress (such as defective cell cycle caused by p53 mutations) or a p53-negative cancer.

[0051] Cancers characterized by p53 mutations or the absence of p53 can be identified, for example, by the methods described in Allred et al., J. Nat. Cancer Institute, Vol. 85, No. 3, 200-206 (1993) and the methods described in the articles listed in the introduction of this application. For instance, the p53 protein can be detected by immunohistochemical methods such as immunostaining.

[0052] Diagnostic tests are typically performed on biological samples selected from the following: tumor biopsy samples, blood samples (isolated and enriched detached tumor cells), stool biopsy, sputum, chromosome analysis, pleural fluid, peritoneal fluid, or urine.

[0053] Besides p53, mutations in other DNA repair factors, such as RAD17, RAD50, and members of the Fanconi anemia complement group, may also predict response to a single Chk1 inhibitor or in combination with chemotherapy. Cancers containing mutations in these DNA repair pathways can be identified by DNA sequence analysis of tumor biopsy tissue or circulating tumor DNA (ctDNA), or, in the case of Fanconi anemia, by assessing DNA foci formation in tumor biopsy specimens using the FANCD2 antibody, as described by Duan et al., Frontiers in Oncology, Vol. 4, 1-8 (2014).

[0054] Therefore, a medically acceptable salt as defined in any of Examples 1.1 to 1.48 may be used to treat members of the following patient subgroups: cancers that have been screened (e.g., by testing biological samples obtained from one or more of these patients) and have been found to have cancers characterized by p53 mutations or p53-negative cancers, or cancers containing RAD17 or RAD50 mutations or mutations of members of the Fanconi anemia complement group.

[0055] Therefore, in other embodiments (Examples 3.24 to 3.30), the present invention provides: 3.24 A pharmaceutically acceptable salt as defined in any of Examples 1.1 to 1.48, used to treat cancer in an individual (e.g., a human individual) who has been screened and identified as having cancer that will be sensitive to treatment with a Chk-1 kinase inhibitor compound or a combination of a chemotherapeutic agent (such as a DNA damaging agent) and a Chk-1 kinase inhibitor compound. 3.25 A pharmaceutically acceptable salt as defined in any of Examples 1.1 to 1.48, used to treat cancer in an individual (e.g., a human individual) who has been screened and identified as having cancer characterized by a defective DNA repair mechanism or a defective cell cycle, such as a defective cell cycle caused by a p53 mutation, or a p53-negative cancer. 3.26 A pharmaceutically acceptable salt as defined in any one of Examples 1.1 to 1.48, for the treatment of cancer in an individual (e.g., a human individual) who has been screened and identified as having cancer characterized by a p53 mutation or a p53-negative cancer, or cancer containing a RAD17 or RAD50 mutation or a mutation of a member of the Fanconi anemia complement group. 3.27 Use of a pharmaceutically acceptable salt as defined in any of Examples 1.1 to 1.48 for the manufacture of a pharmaceutical preparation for the use as defined in any of Examples 3.24 to 3.26. 3.28 A method for treating cancer in an individual (e.g., a human individual) who has been screened and identified as having cancer that is sensitive to treatment with a combination of a Chk-1 kinase inhibitor compound or a chemotherapeutic agent (such as a DNA damaging agent) and a Chk-1 kinase inhibitor compound, the method comprising administering a therapeutically effective amount of a medically acceptable salt as defined in any one of Examples 1.1 to 1.48 and, as appropriate, a chemotherapeutic agent (such as a DNA damaging agent). 3.29 A method for treating cancer in an individual (e.g., a human individual) who has been screened and identified as having cancer, the cancer being characterized by a defective DNA repair mechanism or a defective cell cycle, such as a defective cell cycle caused by a p53 mutation, or a p53-negative cancer, the method comprising administering a therapeutically effective amount of a medically acceptable salt as defined in any one of Examples 1.1 to 1.48. 3.30 A method for treating cancer in an individual (e.g., a human individual) who has been screened and identified as having cancer characterized by a p53 mutation or p53-negative cancer, or cancer containing a RAD17 or RAD50 mutation or a mutation of a member of the Fanconi anemia complement group, the method comprising administering to the individual a therapeutically effective amount of a medically acceptable salt as defined in any one of Examples 1.1 to 1.48. [Combination therapy]

[0056] It is envisioned that medically acceptable salts, as defined in any of Examples 1.1 to 1.48, would be suitable, alone or in combination with chemotherapeutic agents (especially DNA-damaging agents), radiation therapy, or immunotherapy, for the prevention or treatment of a range of proliferative conditions or symptoms. Examples of such conditions and symptoms are described above.

[0057] As defined in any of Examples 1.1 to 1.48, a medically acceptable salt, whether administered alone or in combination with DNA-damaging agents and other anticancer agents and therapies, is generally intended for administration to individuals such as human or animal patients, preferably humans.

[0058] According to another embodiment of the present invention, Example 4.1, a combination of a medically acceptable salt as defined in any one of Examples 1.1 to 1.48 and another chemotherapeutic agent (e.g., an anticancer drug) is provided.

[0059] Examples of chemotherapeutic agents that can be co-administered with a medically acceptable salt as defined in any of Examples 1.1 to 1.48 include: ● Topoisomerase I inhibitors ● Antimetabolites ● Tubulin targeting agents ● DNA binding agents and topoisomerase II inhibitors [ ] ● Alkylating agents [ ] ● Monoclonal Antibodies ● Anti-hormones ● Signal transduction inhibitors [ ] ● Proteasome inhibitors [ ] ● DNA methyltransferase ● Cytokines and retinoids ● Hypoxia-triggered DNA damage agents (e.g., Tirapazamine, TH-302)

[0060] Specific examples of chemotherapeutic agents that can be administered in combination with a medically acceptable salt as defined in any of Examples 1.1 to 1.48 include: Nitrogen mustards, such as dichloromethyldiethylamine, cyclophosphamide, ifosfamide, melphalan, and chlorambucil; Nitrosoururenes, such as carmustine, lomustine, and semustine; Ethyleneimine / methyl melamine compounds, such as triethylene melamine, triethylene thiophosphoramide, and hexamethyl melamine; Alkyl sulfonates, such as busulfan; Three types, such as dacarbazine; Antimetabolites, such as folic acid, methotrexate, trimetrexate, 5-fluorouracil, fludeoxyuridine, gemcitabine, cytosine arabinoside, 5-azacytidine, 2,2'-difluorodeoxycytidine, 6-mercaptopurine, 6-thioguanine, azathioprine, 2'-deoxymyomycin, erythrohydroxynonyl-adenine, fludarabine phosphate, and 2-chlorodeoxyadenosine; Type I topoisomerase inhibitors, such as camptothecin, toponotecan, and irinotecan; Type II topoisomerase inhibitors, such as epipodophyllotoxin (e.g., etoposide and teniposide); Antimitotic drugs, such as paclitaxel, taxotere, vinblastine alkaloids (e.g., vincristine, vinorelbine, vinorelbine), and estramustine (e.g., estramustine phosphate); Antibiotics, such as actimomycin D, daunomycin (rubidomycin), adriamycin, mitoxantrone, idarubicin, bleomycin, mitomycin C, and dactinomycin. Enzymes, such as L-aspartame; Interleukins and biological response modulators, such as interferons (α, β, γ), interleukin-2G-CSF, and GM-CSF: Retinoids, such as retinoic acid derivatives (e.g., bexarotene); Radiation sensitizers, such as metronidazole, misonidazole, demethylmisonidazole, pimonidazole, etanidazole, nimorazole, nicotinamide, 5-bromodeoxyuridine, 5-iododeoxyuridine, and bromodeoxycytidine; Platinum compounds, such as cisplatin, carboplatin, spiroplatin, isopropylplatin, onnaplatin, tetraplatin, and oxaliplatin; Anthraquinones, such as mitoxantrone; Ureas, such as hydroxyurea; Hydrazine derivatives, such as N-methylhydrazine and procarbazine; Adrenocortical inhibitors, such as mitotane and aminoglutethimide; Adrenocortical steroids and antagonists, such as prednisone, dexamethasone, and ampicillin; Progestins, such as hydroxyprogesterone (e.g., hydroxyprogesterone acetate), medroxyprogesterone (e.g., medroxyprogesterone acetate), and megestrol acetate (e.g., megestrol acetate); Estrogens, such as diethylstilbestrol and ethinylestradiol; Anti-estrogens, such as tamoxifen; Androgens, such as testosterone (e.g., testosterone propionate) and fluoromethyltestosterone; Anti-androgens, such as flutamide and leuprolide; Nonsteroidal antiandrogens, such as flutamide; and

[0061] Signal transduction inhibitors, such as PARP inhibitors [e.g., revealed in Cancer Res.; 66: (16)], Mek inhibitors [e.g., revealed in Blood. 2008; 112(6): 2439-2449], farnesyltransferase inhibitors [e.g., revealed in Blood. 2005 Feb 15; 105(4): 1706-16], wee1 inhibitors [e.g., revealed in Haematologica 2014, 99(4): 68], rapamycin and Src inhibitors [e.g., revealed in...] [ Blood ] [.] February 10, 2011; 117(6):1947-57 (as revealed in the paper).

[0062] Immunotherapy agents, such as anti-PD-L1 [as revealed, for example, in Cancer Discov. 2019 (5):646]

[0063] Examples of chemotherapeutic agents that can be used in combination with a pharmaceutically acceptable salt as defined in any of Examples 1.1 to 1.48 include those described in Blasina et al., Mol. Cancer Ther., 2008, 7(8), 2394-2404; Ashwell et al., Clin. Cancer Res., 2008, 14(13), 4032-4037; Ashwell et al., Expert Opin. Investig. Drugs, 2008, 17(9), 1331-1340, Trends in Molecular Medicine, February 2011, Vol. 17, No. 2; and Clin Cancer Res; 16(2), January 15, 2010.

[0064] Specific examples of chemotherapeutic agents that may be used in combination with pharmaceutically acceptable salts as defined in any of Examples 1.1 to 1.48 include antimetabolites (such as capecitabine, cytarabine, fludarabine, gemcitabine, and pemetrezol), topoisomerase-I inhibitors (such as SN38, toponotecan, and irinotecan), platinum compounds (such as carboplatin, oxaliplatin, and cisplatin), topoisomerase-II inhibitors (such as donomycin, cranberry, and etoposide), thymidylate synthase inhibitors (such as 5-fluorouracil), mitotic inhibitors (such as docetaxel, paclitaxel, vincristine, and vinorelbine), and alkylating agents (such as mitomycin C).

[0065] Another group of chemotherapeutic agents that can be used in combination with a pharmaceutically acceptable salt as defined in any of Examples 1.1 to 1.48 includes agents that induce the arrest of replication forks (see Ashwell et al., Clin. Cancer Res., above), and examples of such compounds include gemcitabine, 5-fluorouracil, and hydroxyurea. [Dosimetry]

[0066] A medically acceptable salt as defined in any of Examples 1.1 to 1.48 or a treatment combination as defined in Example 4.1 shall be administered to the patient in need (e.g., human or animal patient) in an amount sufficient to achieve the desired therapeutic effect (e.g., the effect described in Examples 3.1 to 3.30 above).

[0067] A medically acceptable salt as defined in any of Examples 1.1 to 1.48 or a treatment combination as defined in Example 4.1 will generally be administered to an individual who requires such administration, such as a human or animal patient, preferably a human.

[0068] The pharmaceutically acceptable salts as defined in any of Examples 1.1 to 1.48 or the treatment combinations as defined in Example 4.1 will generally be administered in therapeutic or preventative amounts that are generally non-toxic. However, in some cases, the benefit of administering the pharmaceutically acceptable salts of the present invention or the treatment combinations as defined in Example 4.1 may outweigh the disadvantages of any toxicity or side effects, and in such cases, it may be deemed necessary to administer the pharmaceutically acceptable salts of the present invention or the treatment combinations as defined in Example 4.1 in amounts related to the degree of toxicity.

[0069] A medically acceptable salt as defined in any of Examples 1.1 to 1.48, combined with a chemotherapeutic agent or radiotherapy as described and defined above (e.g., as in Example 4.1), may be administered long-term to maintain a beneficial therapeutic effect or may be administered only for a short period. Alternatively, it may be administered in a pulsed or continuous manner.

[0070] A pharmaceutically acceptable salt as defined in any of Examples 1.1 to 1.48, or a treatment combination as defined in Example 4.1, will be administered in an effective amount, i.e., an amount that, alone (as a monotherapy) or in combination with one or more chemotherapeutic agents or radiotherapy, is effective in producing the desired therapeutic effect. For example, an "effective amount" may be the amount of a pharmaceutically acceptable salt that, when administered alone to an individual with cancer or in combination with a DNA-damaging agent or other anticancer agent, slows tumor growth, improves disease symptoms, and / or increases lifespan. More specifically, when used in combination with radiotherapy, a DNA-damaging agent, or other anticancer agent, the effective amount of the pharmaceutically acceptable salt of the present invention is the amount that achieves a greater response when the pharmaceutically acceptable salt is administered in combination with the DNA-damaging agent and / or radiotherapy compared to when the DNA-damaging agent and / or radiotherapy are administered alone. When used as a combination therapy, an "effective amount" of the DNA-damaging agent and / or an "effective" radiation dose is administered to the individual, which is the amount that typically achieves the anticancer effect. As defined in any of Examples 1.1 to 1.48, a pharmaceutically acceptable salt and DNA-damaging anticancer drug may be co-administered to an individual as part of the same pharmaceutical composition, or alternatively as a separate pharmaceutical composition.

[0071] When administered as a standalone pharmaceutical composition, a pharmaceutically acceptable salt as defined in any of Examples 1.1 to 1.48 and a DNA-damaging anticancer drug (and / or radiotherapy) may be administered simultaneously or at different times, subject to the limitation that the enhancing effect of the pharmaceutically acceptable salt as defined in any of Examples 1.1 to 1.48 is retained.

[0072] In one embodiment, a medically acceptable salt as defined in any one of Examples 1.1 to 1.48 is administered before (e.g., up to 8 hours, up to 12 hours, or up to one day prior to) administration of the DNA-damaging anticancer drug.

[0073] In another embodiment, a pharmaceutically acceptable salt as defined in any one of Examples 1.1 to 1.48 is administered after the administration of the DNA-damaging anticancer drug (e.g., at most 8 hours, 12 hours, 24 hours, 30 hours, or 48 hours thereafter). In another embodiment, a first dose of a pharmaceutically acceptable salt as defined in any one of Examples 1.1 to 1.48 is administered one day after the administration of the DNA-damaging anticancer drug, and a second dose of the compound is administered two days after the administration of the DNA-damaging anticancer drug.

[0074] In another embodiment, a first dose of a pharmaceutically acceptable salt as defined in any of Examples 1.1 to 1.48 is administered one day after administration of the DNA-damaging anticancer drug, a second dose of the salt is administered two days after administration of the DNA-damaging anticancer drug, and a third dose of the salt is administered three days after administration of the DNA-damaging anticancer drug.

[0075] Specific dosing regimens for administering pharmaceutically acceptable salts and DNA-damaging anticancer drugs as defined in any of Examples 1.1 to 1.48 can be described as set out in WO2010 / 118390 (Array Biopharma), the contents of which are incorporated herein by reference.

[0076] The dosage of the pharmaceutically acceptable salt of this invention administered to an individual, and (in the case of combination therapy) the DNA-damaging anticancer drug and the radiation dose, will depend on the nature and potency of the DNA-damaging anticancer drug, the type and severity of the disease or symptom, and individual characteristics such as general health, age, sex, weight, and drug tolerance. Those skilled in the art will be able to determine the appropriate dosage based on these and other factors. The effective dosages of commonly used anticancer drugs and radiation therapies are well known to those skilled in the art.

[0077] Whether administered as monotherapy or in combination with a DNA-damaging anticancer drug, the typical daily dose of a pharmaceutically acceptable salt as defined in any of Examples 1.1 to 1.48 may be in the range of 100 picograms to 100 mg / kg body weight, more typically 5 nanograms to 25 mg / kg body weight, and more typically 10 nanograms to 15 mg / kg (e.g., 10 nanograms to 10 mg, and more typically 1 microgram / kg to 20 mg / kg, e.g., 1 microgram to 10 mg / kg) / kg body weight, although higher or lower doses may be administered as needed. The compound may be administered daily or, for example, repeatedly every 2, 3, 4, 5, 6, 7, 10, 14, 21, or 28 days.

[0078] Ultimately, however, the amount of medically acceptable salt used and the type of composition employed will be appropriate to the nature of the disease or physiological condition being treated and will be determined by the physician at his discretion. [Pharmaceutical preparations] [ ]

[0079] As defined in any of Examples 1.1 to 1.48, a pharmaceutically acceptable salt and a treatment combination as defined in Example 4.1 are typically administered to a patient in the form of a pharmaceutical composition. Therefore, in another embodiment of the invention (Example 5.1), the invention provides a pharmaceutical composition comprising a pharmaceutically acceptable salt as defined in any of Examples 1.1 to 1.48, a pharmaceutically acceptable excipient, and, if appropriate, another chemotherapeutic agent.

[0080] In other embodiments, the following are provided: 5.2 The pharmaceutical composition of Example 5.1 comprises about 1% (w / w) to about 95% (w / w) of a pharmaceutically acceptable salt as defined in any one of Examples 1.1 to 1.48 and 99% (w / w) to 5% (w / w) of a pharmaceutically acceptable excipient or combination of excipients and one or more other therapeutically active ingredients as appropriate. 5.3 The pharmaceutical composition of Example 5.2 comprises about 5% (w / w) to about 90% (w / w) of a pharmaceutically acceptable salt as defined in any one of Examples 1.1 to 1.48 and 95% (w / w) to 10% of a pharmaceutical excipient or combination of excipients and, as appropriate, one or more other therapeutically active ingredients. 5.4 The pharmaceutical composition of Example 5.3 comprises about 10% (w / w) to about 90% (w / w) of a pharmaceutically acceptable salt as defined in any one of Examples 1.1 to 1.48 and 90% (w / w) to 10% of a pharmaceutical excipient or combination of excipients. 5.5 The pharmaceutical composition of Example 5.4 comprises about 20% (w / w) to about 90% (w / w) of a pharmaceutically acceptable salt as defined in any one of Examples 1.1 to 1.48 and 80% (w / w) to 10% of a pharmaceutical excipient or combination of excipients. 5.6 The pharmaceutical composition of Example 5.5 comprises about 25% (w / w) to about 80% (w / w) of a pharmaceutically acceptable salt as defined in any one of Examples 1.1 to 1.48 and 75% (w / w) to 20% of a pharmaceutical excipient or combination of excipients.

[0081] The pharmaceutical compositions of the present invention can be in any form suitable for oral, non-enteral, topical, intranasal, intrabronchial, ocular, otolaryngal, rectal, vaginal, or percutaneous administration. When the composition is intended for non-enteral administration, it can be formulated for intravenous, intramuscular, intraperitoneal, subcutaneous administration, or for direct delivery to the target organ or tissue by injection, infusion, or other means of delivery.

[0082] Suitable oral dosage forms include tablets, capsules, granules, pills, lozenges, syrups, solutions, sprays, powders, granules, elixirs and suspensions, sublingual tablets, sprays, powders or patches and buccal patches.

[0083] Therefore, in other embodiments, the present invention provides: 5.7 The pharmaceutical composition of any one of Examples 5.1 to 5.6 is suitable for oral administration. 5.8 The pharmaceutical composition of Example 5.7 is selected from tablets, capsules, granules, pills, lozenges, syrups, solutions, sprays, powders, granules, elixirs and suspensions, sublingual tablets, sprays, powders or patches and buccal patches. 5.9 The pharmaceutical composition of Example 5.8 is selected from tablets and capsules. 5.10 The pharmaceutical composition of any one of Examples 5.1 to 5.6 is suitable for non-enteral administration. 5.11 The pharmaceutical composition of Example 5.10 is formulated for intravenous, intramuscular, intraperitoneal, subcutaneous administration or for direct delivery to a target organ or tissue by injection, infusion or other means of delivery. 5.12 The pharmaceutical composition of Example 5.11 is a solution or suspension for injection or infusion.

[0084] Pharmaceutical compositions containing a pharmaceutically acceptable salt as defined in any of Examples 1.1 to 1.48 (e.g., as defined in any of Examples 5.1 to 5.12) may be formulated according to known techniques, see, for example, Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, PA, USA.

[0085] Therefore, the tablet composition (as in Example 5.9) may contain a unit dose of a pharmaceutically acceptable salt as defined in any of Examples 1.1 to 1.48, and an inert diluent or carrier, such as sugars or sugar alcohols, such as lactose, sucrose, sorbitol, or mannitol; and / or non-sugar-derived diluents, such as sodium carbonate, calcium phosphate, talc, calcium carbonate, or cellulose or its derivatives, such as methylcellulose, ethylcellulose, hydroxypropyl methylcellulose, and starch, such as corn starch. The tablets may also contain standard ingredients, such as binders and granulators (such as polyvinylpyrrolidone), disintegrants (e.g., expandable crosslinking polymers, such as crosslinked carboxymethyl cellulose), lubricants (e.g., stearates), preservatives (e.g., parabens), antioxidants (e.g., BHT), buffers (e.g., phosphate or citrate buffers), and foaming agents (e.g., citrate / bicarbonate mixtures). Such excipients are well known and need not be discussed in detail here.

[0086] The capsule formulation (as in Example 5.9) may be of the form of hard gelatin or soft gelatin and may contain active ingredients in solid, semi-solid, or liquid form. Gelatin capsules may be formed from animal gelatin or its synthetic or plant-derived equivalents.

[0087] Solid dosage forms (e.g., tablets, capsules, etc.) may be coated or uncoated, but are typically coated, such as with a protective film (e.g., wax or varnish) or a release-controlled coating. The coating (e.g., an Eudragit™ type polymer) can be designed to release pharmaceutically acceptable salts at desired locations within the gastrointestinal tract. Therefore, the coating can be selectively degraded under specific pH conditions within the gastrointestinal tract, thereby selectively releasing pharmaceutically acceptable salts into the stomach, ileum, or duodenum.

[0088] As an alternative to or supplement to coating, the drug may be contained in a solid matrix containing a release controller, such as a release delay agent, which is adapted to selectively release a pharmaceutically acceptable salt under varying acidity or alkalinity conditions in the gastrointestinal tract. Alternatively, the matrix material or release-delay coating may be in the form of an erosive polymer (e.g., a maleic anhydride polymer) that is substantially continuously eroded as the dosage form passes through the gastrointestinal tract.

[0089] Topical compositions include ointments, creams, sprays, patches, gels, liquid drops, and inserts (e.g., intraocular inserts). Such compositions can be formulated according to known methods.

[0090] Compositions intended for non-enteral administration (as in Examples 5.10 to 5.12) are typically provided as sterile aqueous or oily solutions or fine suspensions, or as finely pulverized sterile powders for temporary reconstitution with sterile water for injection.

[0091] Examples of preparations for administration rectally or vaginally include pessaries and suppositories, which may be formed, for example, from moldable or waxy materials containing active compounds.

[0092] The composition can be administered by inhalation in the form of an inhalable powder composition, liquid, or powder spray, and can be administered using a standard powder inhaler or aerosol dispensing device. Such devices are well known. For inhalation administration, powdered formulations typically contain pharmaceutically acceptable salts and inert solid powdered diluents, such as lactose.

[0093] Pharmaceutical compositions will generally be presented in unit dosage forms and will therefore typically contain a pharmaceutically acceptable salt sufficient to provide the desired level of biological activity. For example, a pharmaceutical composition intended for oral administration, such as any of Examples 5.1 to 5.9, may contain 2 mg to 200 mg, more typically 10 mg to 100 mg, such as 12.5 mg, 25 mg, and 50 mg of pharmaceutically acceptable salt.

[0094] The pharmaceutical composition may include another chemotherapeutic agent as defined in Example 4.1, depending on the circumstances.

[0095] Therefore, in another embodiment (Example 5.13), the present invention provides a pharmaceutical composition as defined in any one of Examples 5.2 to 5.12, which further comprises another chemotherapeutic agent as defined in Example 4.1. Simple Explanation of the Diagram

[0096] Figure 1 shows the XRPD spectrum of the free base of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxylonitrile (“Compound of Formula (1)”). Figure 2 shows the DSC and TGA traces of the free base of compound (1). Figure 3 shows the GVS curve of the free base of compound (1). Figure 4 shows the XRPD spectra of the free base (top trace) and several crystalline forms of the hydrochloride salt of compound (1). From the second trace at the top to the bottom trace, the crystalline forms of the salt are patterns A, B, C, D and E, respectively. Figure 5 shows the XRPD spectra of the free base (top trace) and several crystalline forms of hydrobromide of compound (1). From the second trace at the top to the bottom trace, the crystalline forms of the salt are patterns A, B, C and D, respectively. Figure 6 shows the XRPD spectra of the free base (top trace) and several crystalline forms of the methanesulfonate of compound (1). From the second trace at the top to the bottom trace, the crystalline forms of the salt are patterns A, B, and C, respectively. Figure 7 shows the XRPD spectra of the free base (top trace) and several forms of L-tartrate of compound (1). From the second trace to the bottom trace, the salt forms are amorphous (second trace downwards), pattern A (third trace downwards), and pattern B (bottom trace) in sequence. Figure 8 shows the XRPD spectra of the free base (top trace) of compound (1) and several crystalline forms of ethanesulfonate. From the second trace to the bottom trace, the crystalline forms of the salt are pattern A and B respectively (the third and fourth traces downwards). Figure 9 shows the XRPD spectra of the free base (top trace) and L-aspartate crystal form (bottom trace) of compound (1). Figure 10 shows the XRPD spectra of several crystalline forms of the benzenesulfonate of compound (1). From top to bottom, the crystalline forms of the salt are patterns A, B, and C. Figure 11 shows the XRPD spectra of several crystalline forms of the toluenesulfonate of compound (1). From top to bottom, the crystalline forms of the salt are patterns A, B, C and D. Figure 12 shows the XRPD spectra of several crystalline forms of the free base and sulfate of compound (1). From top to bottom, the crystalline forms are pattern A (second and third traces downwards) and B (bottom trace) of the free base (top trace) and salt. Figure 13 shows the XRPD spectra of several crystalline forms of the free base and phosphate of compound (1). From top to bottom, the crystalline forms are pattern A (second and third traces downwards) and B (bottom trace) of the free base (top trace) and salt. Figure 14 shows the XRPD spectra of the free base and citrate of compound (1) in several amorphous and crystalline forms. From top to bottom, the traces are the free base (top trace), the amorphous salt (second trace to the bottom), and pattern A and pattern B salts. Figure 15 shows the XRPD spectra of the free base and several crystalline forms of the acetate of compound (1). From top to bottom, the traces are the free base (top trace), salt pattern A, and salt pattern B. Figure 16 shows the XRPD spectra of the free base (top trace) and the pattern A crystalline form (bottom trace) of compound (1). Figure 17 shows the XRPD spectra of several maleate salt crystal forms of compound (1). From top to bottom, these traces are pattern A, pattern B, and pattern C. Figure 18 shows the XRPD spectra of the free base (top trace) of compound (1) and the crystal form of pattern A (middle and bottom traces) of gentianate. Figure 19 shows the XRPD spectra of the free base (top trace) of compound (1) and several crystalline forms of glucuronide (pattern A - middle trace and pattern B - bottom trace). Figure 20 shows the XRPD spectra of the free base (top trace) and several crystalline forms (pattern A - middle trace and pattern B - bottom trace) of compound (1). Figure 21 shows the XRPD spectrum of the naphthalene-2-sulfonate crystal form of compound (1) separated from THF (top trace) and THF:H2O (bottom trace). Figure 22 shows the XRPD spectra of the free base (top trace) of compound (1) and several crystalline forms of oxalate (pattern A - middle trace) and pattern B (bottom trace). Figure 23 shows the XRPD spectra of the free base (top trace) and the crystalline forms A, B, C and D (in descending order starting from the second from the top) of compound (1). Figure 24 shows the XRPD spectra of the free base (top trace) and the crystalline forms D and E (middle and bottom traces) of the sulfate of compound (1). Figure 25 shows the XRPD spectrum of patterned B salt of maleic anhydride. Figure 26 shows the DSC and TGA traces of the patterned B salt of maleic acid ester. Figure 27 shows the XRPD spectrum of patterned A salt of maleic anhydride. Figure 28 shows the DSC and TGA traces of the pattern A salt of maleic acid ester. Figure 29 shows the XRPD spectrum of the patterned C salt of maleic anhydride. Figure 30 shows the DSC and TGA traces of the C salt of maleic acid ester pattern. Figure 31 shows the XRPD spectrum of patterned B salt of malonate. Figure 32 shows the DSC and TGA traces of the malonate pattern B salt. Figure 33 shows the XRPD spectrum of patterned A salt of toluenesulfonate. Figure 34 shows the DSC and TGA traces of toluenesulfonate pattern A salt. Figure 35 shows the XRPD spectrum of the patterned C salt of benzenesulfonate. Figure 36 shows the DSC and TGA traces of the benzenesulfonate pattern of compound (1) C. Figure 37 shows the XRPD spectra of the free base (top trace) and crystalline form of the dimethyl sulfonate of compound (1) as pattern A (middle trace) and pattern B (bottom trace). Figure 38 shows the XRPD spectra of the free base (top trace) and crystalline form of dicis-butenedioate of compound (1) as pattern A (middle trace) and pattern B (bottom trace). Figure 39 shows the XRPD spectra of the free base (top trace) and crystalline form of the dibenzenesulfonate of compound (1) as pattern A (middle trace) and pattern B (bottom trace). Figure 40 shows the XRPD spectra of the free base and various crystalline forms of maleic acid salts. The traces from top to bottom are the free base of compound (1), pattern A mono-maleic acid salt, pattern A di-maleic acid salt, pattern B di-maleic acid salt, and pattern A hemi-maleic acid salt. Figure 41 shows the XRPD spectra of the free base (top trace) and the crystalline form of hemiethane-1,2-disulfonate, pattern A (bottom trace). Figure 42 shows the XRPD spectra of the free base (top trace) and the crystalline form of heminaphthalene-1,5-disulfonate, pattern A (bottom trace). Figure 43 shows the XRPD spectra of the free base and various crystalline forms of hemi-trans-butenedioic acid salts. The traces from top to bottom are the free base of the compound of formula (1), pattern A hemi-trans-butenedioic acid salt, pattern B hemi-trans-butenedioic acid salt, and pattern C hemi-trans-butenedioic acid salt. Figure 44 shows the pattern A of the crystalline form of maleate of compound (1) and its weight vapor phase adsorption (GVS) diagram. Figure 45 shows the GVS diagram of the crystal form of maleate of compound (1). Figure 46 shows the GVS diagram of the crystal form of toluenesulfonate of compound (1). Figure 47 shows the GVS diagram of the crystal form of benzenesulfonate of compound (1). Figure 48 shows the GVS diagram of the crystal form of benzenesulfonate of compound (1). Figure 49 shows the GVS diagram of the crystal form of benzenesulfonate of compound (1). Figure 50 shows the GVS diagram of the crystalline form of naphthalene-2-sulfonate of compound (1). Figure 51 shows the GVS diagram of the malonate of compound (1) in crystal form B. Figure 52 shows the XRPD patterns of various crystalline forms of maleate. From top to bottom, the crystalline forms are pattern A, pattern B, a mixture of A / B, pattern C, pattern D, and pattern E. Figure 53 shows the DVS diagram of the pattern B crystal form of maleic anhydride. Figure 54 shows the DSC and TGA traces of the patterned D salt of maleic acid ester. Figure 55 shows the DSC and TGA traces of the patterned E salt of maleic acid ester. Implementation

[0097] [Example] [ ] [Analysis Methods] [ ] [Proton] [-NMR] [ ] Salt formation (by observing the proton shift relative to the free base) and identification of the salt as a 1:1 (free base: acid molar ratio) stoichiometric salt were confirmed by their 1H NMR spectra, which were collected using a JEOL ECX 400MHz spectrometer equipped with an autosampler. Samples were dissolved in suitable deuterated solvents for analysis. Data were acquired using Delta NMR processing and control software version 4.3. [X] [Powder diffraction] [(XRPD)] [ ]

[0098] X-ray powder diffraction patterns were collected using a PANalytical diffractometer with Cu Kα radiation (45 kV, 40 mA), an θ-θ goniometer, a focusing lens, a diverging slit (1 / 2''), a Soler slit under both incident and diverging beams (4 mm), and a PIXcel detector. X'Pert Data Collector, version 2.2f, was used for data collection, and X'Pert Data Viewer, version 1.2d, was used to display the data. XRPD patterns were obtained under ambient conditions using PANalytical X'Pert PRO via a transmission foil stage (polyimide-Kapton, 12.7 µm thick film). The data collection range was 2.994 – 35º2θ, with a continuous scan rate of 0.202004ºs⁻¹. Differential scanning calorimetry [(DSC)] [ ]

[0099] DSC data were collected on a PerkinElmer Pyris 6000 DSC equipped with a 45-position sample holder. Energy and temperature calibration were performed using a calibrated indium assay instrument. A predetermined sample volume (0.5–3.0 mg) was placed in a pinhole aluminum dish and heated at 20 °C / min from 30 °C to 350 °C, or as required by the experiment. The sample was purged with 20 mL / min of dry nitrogen. Instrument control, data collection, and analysis were performed using Pyris Software v11.1.1 (Revision H). [Pyrolysis Gravimetric Analysis] [(TGA)] [ ]

[0100] TGA data were collected on a Perkin Elmer Pyris 1 TGA equipped with a 20-position autosampler. The instrument was calibrated using certified weights and temperature-calibrated using certified Alumel and Perkalloy instruments. Pre-weighed sample volumes (1–5 mg) were loaded onto pre-weighed aluminum crucibles and heated from ambient temperature to 400 °C at 20 °C / min. The sample was purged with nitrogen at 20 mL / min. Instrument control, data collection, and analysis were performed using Pyris Software v11.1.1 (Revision H). [Gravity gas phase adsorption] (GVS) [ ]

[0101] The salt of the present invention was subjected to GVS studies using the method described below: Adsorption isotherms were obtained using a Hiden Isochema moisture adsorption analyzer (model IGASorp) controlled by IGASorp system software V6.50.48. The sample temperature was maintained at a constant temperature (25°C) using the instrument controls. Humidity was controlled by a mixed flow of drying and humidifying nitrogen gas at a total flow rate of 250 ml / min. The relative humidity (RH) content of the instrument was verified by measuring three calibration Rotronic salt solutions (10-50-88%). The weight change of the sample with humidity was monitored using a microbalance (accuracy + / - 0.005 mg). Defined amounts of sample were placed in a weighing mesh stainless steel basket under ambient conditions. A complete experimental cycle typically consisted of three scans (adsorption, desorption, and adsorption) at constant temperature (25°C) and 10% RH intervals (60 minutes at each humidity level) within the range of 0-90%. This type of experiment should demonstrate the ability of the sample under study to absorb (or not absorb) moisture within a very defined range of humidity. [HPLC] [method] [1]

[0102] HPLC analysis was performed on an Agilent 1110 series HPLC system. The column used was Aquity BEH Phenyl; 30 × 4.6 mm, 1.7 μm particle size (Ex Waters, PN: 186004644). The flow rate was 2.0 mL / min. Mobile phase A was water:trifluoroacetic acid (100:0.03%) and mobile phase B was acetonitrile:trifluoroacetic acid (100:0.03%). Detection was performed using 210 nm UV. The injection volume was 5 μL and the following gradient was used: time %A %B 0 95 5 5.2 5 95 5.7 5 95 5.8 95 5 6.2 95 5 [HPLC] [method] [2]

[0103] HPLC analysis was performed on an Agilent 1110 / 1200 series HPLC system. The column used was a Triart C18; 150 × 4.6 mm, 3.0 μm particle size (Ex Waters, PN: 186004644). The flow rate was 1.0 mL / min. Mobile phase A was water:trifluoroacetic acid (100:0.1%) and mobile phase B was acetonitrile:trifluoroacetic acid (100:0.1%). Detection was performed using 302 nm UV. The injection volume was 5 μL, the column temperature was 40 °C, and the following gradient was used: Time (min) %A %B 0 95 5 5 65 35 10 65 35 18 5 95 22.5 5 95 twenty three 95 5 [Example] [1] [5-[[5-[4-(4-)] [fluorine] [-1-] [methyl] [-4-] [piperidinyl] [)-2-] [Methoxy] [-] [Phenyl] []-1H-] [Pyrazole] [-3-] [base] []] [Amine] []] [Pyr] [𠯤] [-2-] [Preparation and Characterization of Formonitrile Free Base]

[0104] The title compound was prepared by the method of Example 64, Method L of WO 2015 / 20390 (the contents of which are incorporated herein by reference), but the compound was isolated as a free base rather than a hydrochloride. The free base was characterized by X-ray powder diffraction (XRPD), differential scanning calorimetry (DSC), and pyrolysis gravimetric analysis (TGA). The XRPD spectra and DSC and TGA traces are shown in Figures 1 and 2.

[0105] The free base is crystalline according to XRPD. DSC thermal imaging shows the primary melting endothermic process with an initial temperature of 205.6°C and a peak temperature of 214°C. TGA thermal imaging shows a weight loss of up to 2.8% at 150°C. The 1H NMR spectrum of the solid conforms to the molecular structure. Since there is no significant solvent in the NMR spectrum, the weight loss shown in the TGA thermal image is related to water loss during heating of the material.

[0106] The GVS curve for the free alkali is shown in Figure 3. During the initial desorption cycle, the solid lost 2 wt% of water from 50% relative humidity (RH) to 0% RH. In subsequent adsorption cycles, the solid gained 8% water up to 90% RH. The water absorption is reversible, and lag is noted. The theoretical water content of the free alkali monohydrate is 4.2%, therefore, water is absorbed to the dihydrate level at extreme humidity. [Example] [2] [ ] [Preparation of Salt] [Small-scale approach] [ ]

[0107] The acid addition salt of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxynitrile is prepared from a free base by the following small-scale methods 1 to 7. [method] [1]: THF-mediated

[0108] The free base (50 mg) of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxynitrile was packed into 16 crystallizer tubes. THF (2 mL, 40 volumes) was added, and the resulting mixture was heated to 60°C. Acid (1 M, 1 equivalent) was packed into individual aliquots. The solutions were held at a constant temperature and equilibrated for 1 hour. The solutions were then cooled to room temperature and equilibrated for 18 hours, followed by filtration and drying under vacuum for 18 hours. Where crystallization did not occur, the samples were further processed by removing the solvent using nitrogen and grinding the solid with MeOH. The following salts required solvent reduction and grinding: ethanesulfonate, benzenesulfonate, acetate, and malonate. [method] [2]:THF:MeCN mediated

[0109] Method 2 is the same as Method 1, except that a mixture of THF:MeCN (1:1) (1 mL, 20 volumes) is used as the solvent and the mixture is heated to 50°C. Benzenesulfonates require solvent reduction and grinding. [method] [3]:THF:Water-mediated

[0110] Method 3 is the same as Method 2, except that THF:water (95:5) (1 mL, 20 volumes) is used as the solvent and the mixture is heated to 50°C. Benzenesulfonic acid, acetic acid, L-glutamic acid, and L-aspartate require solvent reduction and grinding. [method] [4]: Using excess acid via THF-mediated

[0111] Method 4 is the same as Method 1, except that the acid (1M, 1.84 equivalents) is loaded into a single aliquot. This method is used to separate the following salts: hydrochloride pattern A, hemi-trans-butenedioate pattern A, hydrobromide pattern C, dimethyl sulfonate pattern A, dicis-butenedioate pattern A, dibenzene sulfonate pattern A, toluene sulfonate pattern C, and acetate pattern B. [method] [5]: Use of excessive acid via THF:water mediated

[0112] Method 5 is the same as Method 2, except that the acid (1M, 1.84 equivalents) is loaded into a single aliquot. This method is used to separate the following salts: L-tartrate pattern B, toluenesulfonate pattern A, phosphate pattern B, citrate pattern B, acetate pattern B, L-glutamate pattern A, hydrochloride pattern D, hydrobromide pattern D, dimethylsulfonate pattern B, dicis-butenedioate pattern B, benzenesulfonate pattern B, and sulfate pattern C. [method] [6]: Using excess acid via THF-mediated

[0113] Method 6 is the same as Method 1, except that the free base (30 mg) and acid (1 M, 2 eq) are loaded into individual aliquots. This method is used to prepare patterned B salts of hydrochloride. [method] [7]: Using 0.5 equivalent acid via THF-mediated...

[0114] Method 7 is the same as Method 1, except that 0.5 equivalents of acid are added in each case. Method 7 is used to separate the following salts: hemibutenedioate pattern A and hemisulfate pattern A, hemiethane-1,2-disulfonate pattern A and heminaphthyl-1,5-disulfonate pattern A. [5-[[5-[4-(4-)] [fluorine] [-1-] [methyl] [-4-] [piperidinyl] [)-2-] [Methoxy] [-] [Phenyl] []-1H-] [Pyrazole] [-3-] [base] []] [Amine] []] [Pyr] [𠯤] [-2-] [Medium-scale preparation of formonitrile salts] [ ] [Medium-scale approach] [1]

[0115] Using conditions similar to those used in small-scale Method 1, but with 300 mg of free base for larger-scale salt preparation. The following salts were prepared in this manner. • Maleate pattern C, thermal cycling at >200℃ yields toluenesulfonate pattern D. • Maleate pattern A, adjusted at 40℃ / 75%RH to obtain maleate pattern B • Benzenesulfonate pattern B • Pattern A of naphthyl-2-sulfonate

[0116] This method is modified by using 100 mg of free base to form: • Oxalate pattern A [Medium-scale approach] [2]

[0117] The preparation of malonate pattern B was carried out by scaling up the scale using the following method: 300 mg of free base of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxynitrile was weighed into a 25 mL round-bottom flask. THF:water (95:5, 20 volumes) was added, and the mixture was equilibrated at 60 °C for 15 minutes. Malonic acid (1 equivalent) was added, and the mixture was equilibrated at 60 °C for 15 minutes. The mixture was cooled to room temperature and equilibrated for 30 minutes. The mixture was then rapidly evaporated using a rotary evaporator at 50 °C and 210 rpm. This produced a beige powder, which was then matured in MeOH (20 volumes) at room temperature for 18 hours. The resulting suspension was separated by vacuum filtration, and the solids were vacuum dried at 45 °C over the weekend. [Medium-scale approach] [3]

[0118] The method uses the conditions described in small-scale method 5, except that it uses 300 mg of free base and 2 equivalents of acid. This method is used to prepare pattern A salt of bis(cis-butenedioate). [Medium-scale approach] [4]

[0119] The method uses the conditions described in small-scale method 4, except that it uses 300 mg of free base and 2 equivalents of acid. This method is used to prepare bis(benzenesulfonate) pattern B. [Mature Methodology] [ ]

[0120] The benzenesulfonate pattern C is formed after the benzenesulfonate pattern B has matured in water (24 hours).

[0121] The maleate pattern C is formed after the maleate B has matured in water (24 hours).

[0122] The table below provides an overview of the methods used to prepare salt and the physical appearance of the salts prepared therefrom. [surface] [-] [Salt Preparation] [ ] [Salt] [method] [Appearance] Hydrochloride (XRPD pattern A) twenty four Light yellow solid Hydrochloride (XRPD pattern B) 6 grayish-white solid Hydrochloride (XRPD pattern C) 1 grayish-white solid Hydrochloride (XRPD pattern D) 5 grayish-white solid Hydrochloride (XRPD pattern E) 3 pale yellow solid Hydrobromide (XRPD pattern A) 1, 3 pale yellow solid Hydrobromide (XRPD pattern B) 2 grayish-white solid Hydrobromide (XRPD pattern C) 4 grayish-white solid Hydrobromide (XRPD pattern D) 5 grayish-white solid Methanesulfonate (XRPD pattern A) 1 grayish-white solid Methanesulfonate (XRPD pattern B) 2 grayish-white solid Methanesulfonate (XRPD pattern C) 3 grayish-white solid L-Tartrate (XRPD Pattern A) 2 grayish-white solid L-Tartrate (XRPD Pattern B) 3, 5 grayish-white solid Ethyl sulfonate (XRPD pattern A) 1 grayish-white solid Ethyl sulfonate (XRPD pattern B) twenty three grayish-white solid L-Aspartate (XRPD pattern A) 3 grayish-white solid Benzenesulfonate (XRPD pattern A) 1, 2, 3 grayish-white solid Benzenesulfonate (XRPD pattern B) 5, 1 (proportionally increased) grayish-white solid Benzenesulfonate (XRPD pattern C) Water maturation of benzenesulfonate B Toluenesulfonate (XRPD pattern A) 1, 3, 5 grayish-white solid Toluenesulfonate (XRPD pattern B) 2 grayish-white solid Toluenesulfonate (XRPD pattern C) 1 (proportionally increased), 4 grayish-white solid Toluenesulfonate (XRPD pattern D) Heat cycle of pattern C Sulfate (XRPD pattern A) 1, 2 Light yellow solid Sulfate (XRPD pattern B) 3 Light yellow solid Sulfate (XRPD pattern C) 5 grayish-white solid Sulfate (XRPD pattern D) 7 grayish-white solid Sulfate (XRPD pattern E) 7 grayish-white solid Phosphate (XRPD pattern A) 1, 2 grayish-white solid Phosphate (XRPD pattern B) 3, 5 grayish-white solid Citrate (XRPD pattern A) 2 grayish-white solid Citrate (XRPD pattern B) 3, 5 grayish-white solid Acetate (XRPD pattern A) 1, 3 grayish-white solid Acetate (XRPD pattern B) 5 grayish-white solid L-Glutamate (XRPD Pattern A) 3, 5 grayish-white solid maleic acid salt (XRPD pattern A) 1, 2, 3 grayish-white solid maleic acid salt (XRPD pattern B) Adjusting maleate A at 40℃ / 75%RH Light brown to yellow solid maleic acid salt (XRPD pattern C) Water maturation of maleate B Light brown to yellow solid Gentianate (XRPD pattern A) 1, 3 grayish-white solid Glucuronite (XRPD pattern A) 1 Light yellow solid Glucuronite (XRPD pattern B) 3 Light yellow solid Malonate (XRPD pattern A) 1 grayish-white solid Malonate (XRPD pattern B) 3 grayish-white solid Naphthyl-2-sulfonate (XRPD pattern A) 1, 3 grayish-white solid Oxalate (XRPD pattern A) 1 grayish-white solid Oxalate (XRPD pattern B) 3 grayish-white solid

[0123] The characteristic data of the salts prepared according to the method described above are presented in the table below. [ ] [ ] [surface] [-] [Characteristics of Salt] [ ] [Salt] [XRPD] [picture] [Thermal Profile] [DSC / TGA] [NMR] hydrochloride (XRPD Pattern A) Figure 4 Second trace from the top DSC events: endothermic at 169℃ and 285℃, exothermic at 219℃ 1H NMR confirmed salt formation and confirmed the IC50 required for stoichiometry. hydrochloride (XRPD pattern B) Figure 4. Third trace from the top DSC event: widespread heat absorption at 220℃ 1H NMR confirmed salt formation and confirmed the IC50 required for stoichiometry. hydrochloride (XRPD pattern C) Figure 4. Third trace from the bottom DSC events: heat absorption at 172℃ and 209℃ 1H NMR confirmed salt formation and confirmed the IC50 required for stoichiometry. hydrochloride (XRPD pattern D) (Prepared using excess acid) Figure 4. Second trace from the bottom DSC events: minor heat absorption at 140 and 214°C TGA event: At most 3.8% loss at 80℃, followed by a 3.9% loss, consistent with the first endothermic event. 1H NMR confirmed salt formation and confirmed the IC50 required for stoichiometry. hydrochloride (XRPD pattern E) Figure 4 Bottom trace DSC events: 143, 174, 196, and 217°C (primary melting point) TGA incident: Loss of up to 3.1% at 150℃ 1H NMR confirmed salt formation and confirmed the IC50 required for stoichiometry. Hydrobromide (XRPD pattern A) Figure 5 Second trace from the top DSC event: Initial temperature 205℃, peak temperature 209℃ TGA event: Up to 2.3% loss at 100°C, and 3.6% loss at the main melting point. 1H NMR confirmed salt formation and confirmed the IC50 required for stoichiometry. Hydrobromide (XRPD pattern B) Figure 5 shows the third trace from the top. DSC event: slight heat absorption at 180°C followed by 209°C and widespread heat absorption at 270°C. TGA event: Up to 4.7% loss at 100°C and 3.7% loss during the second endothermic period. 1H NMR confirmed salt formation and confirmed the IC50 required for stoichiometry. Hydrobromide (XRPD pattern C) (Prepared using excess acid) Figure 5. Second trace from the bottom DSC Event: Shoulder Edge at 247℃ TGA incident: Loss of up to 4.4% at 150℃ 1H NMR confirmed salt formation and confirmed the IC50 required for stoichiometry. Hydrobromide (XRPD pattern D) (Prepared using excess acid) Figure 5 Bottom trace DSC event: widespread heat absorption at 252℃ TGA incident: Loss of up to 7.3% at 150℃ 1H NMR confirmed salt formation and confirmed the IC50 required for stoichiometry. Methanesulfonate (XRPD pattern A) Figure 6 Second trace from the top DSC event: Peak temperatures at 195 and 209℃ TGA event: Up to 1.6% loss at 100℃ and 4.6% loss within the main endothermic temperature range of 150-250℃. 1H NMR confirms single stoichiometry Methanesulfonate (XRPD pattern B) Figure 6. Second trace from the bottom DSC event: Peaks at 164 and 210℃ TGA events: Losses of up to 4.2% at 100°C, followed by 2.25% and 2.9%, consistent with endothermic reactions. 1H NMR confirms single stoichiometry Methanesulfonate (XRPD pattern C) Figure 6 Bottom trace DSC Events: Peak Temperatures at 171℃ and 200℃ TGA event: 4.6% loss at up to 150°C, followed by a 2.8% loss between 200 and 250°C. 1H NMR confirms single stoichiometry L-Tartrate (XRPD Pattern A) The two traces in the middle of Figure 7 DSC event: trace amounts at 145℃, widespread endothermic events at 200℃. TGA event: Loss of up to 1.6% at 100°C and 0.8% between 175 and 225°C. 1H NMR confirms single stoichiometry L-Tartrate (XRPD Pattern B) Figure 7 Bottom trace DSC event: trace amounts at 155℃, widespread endothermic events at 200℃. TGA incident: Loss of up to 4.3% at 150℃ 1H NMR confirms single stoichiometry Ethyl sulfonate (XRPD pattern A) Figure 8. Second trace from the top DCS events: 145℃ and 215℃ TGA event: At most 1.8% loss at 100°C, followed by a 2.4% loss in the first endothermic phase, and a 2.8% loss in the second endothermic phase. 1H NMR confirms single stoichiometry Ethyl sulfonate (XRPD pattern B) Figure 8 shows the two traces at the bottom. DSC event: Widespread heat absorption at 153 and 192℃ TGA incident: Loss of up to 4.8% at 100℃, and loss of up to 1.4% at 175℃. 1H NMR confirms single stoichiometry L-Aspartate (XRPD pattern A) Figure 9 Bottom trace DSC events: endothermic at 106 and 163°C, bimodal at 207 and 220°C. TGA incident: Loss of up to 6.9% at 100℃, followed by a 2.4% loss up to 175℃. 1H NMR confirms single stoichiometry Benzenesulfonate (XRPD pattern A) Figure 10 Top trace DSC events: heat absorption at 166, 189, and 217°C TGA event: Up to 0.8% loss at 100°C and 3% loss during the second endothermic reaction. 1H NMR confirms single stoichiometry Benzenesulfonate (XRPD pattern B) Figure 10 Middle trace DSC event: Bimodal endothermic reaction, with peaks at 211 and 223℃. TGA event: 0.4% starting from 130°C before the main melting point. 1H NMR confirms single stoichiometry Benzenesulfonate (XRPD pattern C) Figure 10 Bottom trace DSC event: Single heat absorption at 230℃ TGA incident: Loss of up to 2.1% at 100℃ 1H NMR confirms single stoichiometry Toluenesulfonate (XRPD pattern A) Figure 11 Top trace DSC event: 106℃, followed by the main melting point of 234℃. TGA incident: Loss of up to 2.7% at 100℃ 1H NMR confirms single stoichiometry Toluenesulfonate (XRPD pattern B) Figure 11 Second trace from the top DSC event: Widespread heat absorption at 157℃ and 217℃ TGA event: At most 2.6% loss at 100℃, followed by losses of 1.6% and 2.8%, consistent with endothermic reactions. 1H NMR confirms single stoichiometry Toluenesulfonate (XRPD pattern C) Figure 11 Second trace from the bottom DSC events: shoulder endothermic events at 125°C (endothermic), 185°C (exothermic), and 225°C (exothermic). TGA event: Up to 1.5% loss at 150℃ and 1.6% loss at 100-175℃. 1H NMR confirms single stoichiometry Toluenesulfonate (XRPD pattern D) Figure 11 Bottom trace DSC Event: Single Primary Melting Point at 222℃ TGA incident: No mass loss before the main melting point 1H NMR confirms single stoichiometry Sulfate (XRPD pattern A) The two traces in the middle of Figure 12 DSC event: heat absorption at 187℃ and 267℃, heat release at 250℃ TGA event: Up to 3.4% loss at 150°C, and 3.3% loss during the first endothermic reaction. 1H NMR confirmed salt formation and confirmed the IC50 required for stoichiometry. Sulfate (XRPD pattern B) Figure 12 Bottom trace DSC events: Widespread endothermic at 119°C, 174°C, and 265°C, exothermic at 250°C. TGA event: up to 3.9% loss at 110°C, followed by a 2.7% loss before exothermic reaction. 1H NMR confirmed salt formation and confirmed the IC50 required for stoichiometry. Sulfate (XRPD pattern C) (Prepared from excess acid) Figure 23 Middle trace DSC events: broad peaks at 131℃ and 179℃, and rapid exothermic reaction at 272℃. TGA incident: Loss of up to 9.2% at 125℃ 1H NMR confirmed salt formation and confirmed the IC50 required for stoichiometry. Sulfate (XRPD pattern D) (Prepared from 0.5 equivalents of acid, stoichiometry not confirmed, marked as hemisulfate pattern A in the report) Figure 23 Bottom trace DSC event: Single heat absorption at 187℃ TGA incident: Up to 0.7% mass loss at 100°C 1H NMR confirmed salt formation and confirmed the IC50 required for stoichiometry. Sulfate (XRPD pattern E) (Prepared from 0.5 equimolar acid but IC indicates a single salt, labeled as hemisulfate pattern A in the report) Figure 24 Bottom trace DSC event: slight endothermic reaction at 201℃ TGA event: 0.34% loss before melting and endothermic reaction. 1H NMR confirmed salt formation, and IC confirmed monostoichiometry. Phosphate (XRPD pattern A) The two traces in the middle of Figure 13 DSC event: Heat absorption 164℃ TGA: 2.1% loss before the main melting point, followed by a 10% step loss. NMR is uncertain; IC is needed to determine the stoichiometry. Phosphate (XRPD pattern B) Figure 13 Bottom trace DSC event: heat absorption at 153 and 203°C TGA incident: Loss of up to 3.6% at 150℃ 1H NMR confirms salt formation; IC is needed to confirm stoichiometry. Citrate (XRPD pattern A) Figure 14 Second trace from the bottom DSC Events: Heat Endothermation of 163 TGA Events: Up to 1.5% Loss at 100°C 1H NMR confirms single stoichiometry Citrate (XRPD pattern B) Figure 14 Bottom trace DSC event: Bimodal endothermic peaks at 117 and 139 °C and widespread endothermic effect at 192 °C. TGA event: at most 3.1% loss at 100℃, followed by a 1.6% loss, consistent with the first endothermic event. 1H NMR confirms single stoichiometry Acetate (XRPD pattern A) Figure 15 Middle trace DSC event: endothermic shoulder at 131°C, followed by an event at 213°C. 1H NMR confirms single stoichiometry Acetate (XRPD pattern B) Figure 15 Bottom trace DSC events: 100℃, 156℃ TGA event: Loss of up to 6.2% at 100℃, and 11.5% at 100-165℃. 1H NMR confirms single stoichiometry L-Glutamate (XRPD Pattern A) Figure 16 Bottom trace DSC events: 96, 151, 169, and 201℃ TGA incident: Mass loss at up to 110°C and 3.3% loss at 150-200°C. 1H NMR confirms single stoichiometry maleic acid salt (XRPD pattern A) Figure 17 Top trace and Figure 27 DSC and TGA - Figure 28 DSC: The main melting point of the peak at 201℃ TGA event: No loss before the main melting point. 1H NMR confirms single stoichiometry maleic acid salt (XRPD pattern B) Figure 17 shows the intermediate trace and Figure 25 shows the intermediate trace. DSC and TGA - Figure 26 DSC: The main melting point at the peak of 201℃ 1H NMR confirms single stoichiometry maleic acid salt (XRPD pattern C) Figure 17 shows the bottom trace and Figure 29 shows the bottom trace. DSC and TGA - Figure 30 DSC event: Peak at the main melting point of 202℃ TGA incident: Loss of up to 1.2% at 120℃ 1H NMR confirms single stoichiometry Gentianate (XRPD pattern A) Figure 18 Middle and bottom traces DSC event: Endothermic reaction at the acromion at 181°C TGA event: Up to 0.4% loss at 100℃ and 0.25% loss between 100-165℃. 1H NMR confirms single stoichiometry Glucuronite (XRPD pattern A) Figure 19 Intermediate trace DSC event: Single endothermic peak at 166℃ TGA incident: Loss of up to 0.3% at 100℃ 1H NMR confirms single stoichiometry Glucuronite (XRPD pattern B) Figure 19 Bottom trace DSC event: Single endothermic peak at 159℃ TGA event: Up to 2.6% loss at 100℃ and 1% loss at 100-130℃. 1H NMR confirms single stoichiometry Malonate (XRPD pattern A) Figure 20 Middle trace DSC event: Single endothermic peak at 140℃ TGA incident: Loss of up to 0.5% at 100℃ 1H NMR confirms single stoichiometry Malonate (XRPD pattern B) Figure 20 Bottom trace DSC event: Single endothermic peak at 165℃ TGA incident: Loss of up to 0.5% at 100℃ 1H NMR confirms single stoichiometry Naphthyl-2-sulfonate (XRPD pattern A) Figure 21 Two traces DSC event: Peak at 243℃, high melting endothermic temperature. TGA incident: Loss of up to 0.8% at 100℃ 1H NMR confirms single stoichiometry Oxalate (XRPD pattern A) Figure 22 Middle trace DSC event: The peak is an endothermic event at 200℃. TGA incident: Loss of up to 0.1% at 100℃ 1H NMR confirmed salt formation and confirmed the IC50 required for stoichiometry. Oxalate (XRPD pattern B) Figure 22 Bottom trace DSC event: widespread heat absorption at 190℃ TGA incident: Up to 1.6% loss at 100℃ and 1.6% loss at 120-170℃. 1H NMR confirmed salt formation and confirmed the IC50 required for stoichiometry.

[0124] The characteristics of the disalts and hemisalts prepared using the methods described above are also described below. [Double Salt] [ ] [Salt] [XRPD] [picture] [Thermal Profile] [DSC / TGA] [NMR] [Preparation Method] [Appearance] dimethylsulfonate (XRPD Pattern A) Figure 37 Middle Trace Line DSC event: Single endothermic peak at 173℃ TGA incident: Loss of up to 4.3% at 125℃ 1H NMR confirms salt formation and dual stoichiometry 4 grayish-white powder dimethylsulfonate (XRPD pattern B) Figure 37 Bottom trace DSC event: The peaks at 132 and 146 °C show double endothermic activity, followed by endothermic activity at 169 °C. TGA event: up to 3.4% loss at 80°C, followed by a 2.9% loss during the bimodal endothermic phase. 1H NMR confirms salt formation and dual stoichiometry 4 grayish-white powder Dicis-butenedioate (XRPD pattern A dicis-butenedioate) Figure 38 Middle Trace Line DSC event: endothermic melting at 185°C TGA incident: Loss of up to 1.5% at 100℃ 1H NMR indication 1:1.75 stoichiometry (Hybrid mono / dual phase) 4 grayish-white powder Dicis-butenedioate (XRPD pattern B dicis-butenedioate) Figure 38 Bottom trace DSC event: endothermic melting at 191℃ TGA incident: Loss of up to 1.3% at 100℃ 1H NMR confirms salt formation and dual stoichiometry 4 grayish-white powder Bisbenzenesulfonate (XRPD pattern A) Figure 39 Intermediate trace DSC event: Single heat absorption at 212℃ TGA incident: Loss of up to 0.8% at 100℃ 1H NMR confirms salt formation and dual stoichiometry 4 grayish-white powder Dibenzenesulfonate (XRPD pattern B) Figure 39 Bottom trace DSC event: Single rapid heat absorption at 217°C TGA event: 0.25% loss from 130℃ to 200℃. 1H NMR confirms salt formation and dual stoichiometry 4 (proportionally enlarged) grayish-white powder [Half-salt] [ ] [Salt] [XRPD] [picture] [Thermal Profile] [DSC / TGA] [NMR] [Preparation Method] [Appearance] hemibutenedioic acid salt (XRPD Pattern A) Figure 40 Bottom trace DSC Event: Primary Melting Point at 192℃ TGA incident: Loss of up to 0.2% at 100℃ 1H NMR confirmed salt formation and semi-stoichiometry. THF solvates 7 grayish-white solid Semi-ethane-1,2-disulfonate (XRPD pattern A) Figure 41 Bottom trace DSC event: Endothermic reaction at the acromion at 196°C TGA event: Up to 1.65% loss at 100°C and 2.1% loss before the main melting point. 1H NMR confirmed salt formation and semi-stoichiometry 7 grayish-white solid Heminaphthalene-1,5-disulfonate (XRPD pattern A) Figure 42 Bottom trace DSC event: a small amount of heat release at 208℃, and a large amount of heat absorption at 262℃. TGA event: Up to 1.4% loss at 100°C and 5.3% loss before the main melting point. 1H NMR confirmed salt formation and semi-stoichiometry 7 grayish-white solid Hemibutenedioic acid salt (XRPD pattern A) Figure 43 The second trace from the top DSC events: heat absorption at 172 and 217°C TGA event: 7.2% loss before heat absorption and then another 4.9% loss during the second heat absorption. 1H NMR confirmed salt formation and semi-stoichiometry 1 grayish-white solid Hemibutenedioic acid salt (XRPD pattern B) Figure 43 The second trace from the bottom DSC event: minor heat absorption at 167℃, widespread heat absorption at 223℃. TGA event: Loss of up to 4.5% at 100℃, followed by a 2.7% loss at 167℃. 1H NMR confirmed salt formation and semi-stoichiometry 2 grayish-white solid Hemibutenedioic acid salt (XRPD pattern C) Figure 43 Bottom trace DSC event: Single heat absorption at 182℃ TGA incident: Loss of up to 2.7% at 100℃ 1H NMR confirmed salt formation and semi-stoichiometry 3 grayish-white solid [Example] [3A] [Determination of the solubility of salt in water] [ ]

[0125] The free base of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxynitrile and the selected salt (30 mg) were weighed into a crystallizer, filled with water for injection (WFI) (1 mL), and the sample was allowed to equilibrate for 24 hours (25°C). The solid was separated by vacuum filtration, and the filtrate was used to evaluate the solubility using HPLC (HPLC Method 1). Salt [Water solubility mg / mL 24 h] Toluenesulfonate pattern C 1.55 maleic acid ester pattern B 1.4 Dicis-butenedioate pattern B 0.15 Sulfate pattern D 1.6 Benzenesulfonate pattern B 1.18 Bisbenzenesulfonate pattern B 0.06 Free base 0.39 Malonate pattern B 7.41 Oxalate Pattern A 3.16 Hydrochloride pattern C 10.37 [in conclusion] [ ]

[0126] A large number of crystalline salts were identified, but most of them showed a tendency to hydrate / solvate and had complex thermal profiles. Hydrochlorides are known to exhibit polymorphism and poor thermal profiles indicating hydration / solvation in International Patent Application WO 2015 / 20390, and are therefore not considered as preferred candidates for the preparation of solid formulations.

[0127] Toluenesulfonate, maleate, benzenesulfonate, malonate, and oxalate all showed superior solubility compared to the free base, but oxalate has lower crystallinity and is therefore not considered for further development. The disalts disproportionate in water and are therefore not considered candidates for further development. [Example] [3B] [Determination of the solubility of salts in biologically relevant media] [ ] experiment:

[0128] Weigh the free base of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxynitrile and the selected salt (30 mg) into a crystallizer tube. Add biological media (DI H2O, FeSSIF, FaSSIF, and FaSSGF) (2 mL). Allow the sample to equilibrate over 24 hours (25°C). Perform solubility determination using HPLC (see Method 1 above). [Salt form] [FeSSIF (pH 5.14)] [FaSSIF (pH 6.71)] [FaSSGF (pH 1.37)] Free base 0.6 0.04 2.93 maleic acid ester pattern B 0.57 0.07 3.97 Benzenesulfonate pattern B 0.57 0.04 0.55 maleic acid ester pattern C 0.61 0.04 0.32 Hydrochloride pattern C 0.61 0.15 8.39 Malonate pattern B 0.27 0 3.24 FaSSIF: Simulated intestinal fluid during fasting FeSSIF: Simulated intestinal fluid during feeding FaSSGF: Simulated gastric juices during fasting

[0129] Biorelevant solubility assessments of the free base and selected salts showed overall poor solubility of less than 1 mg / mL. As a general trend, increasing solubility was observed from FaSSIF to FeSSIF and then to FaSSGF. In FaSSGF, maleate and malonate showed better solubility than the free base. Most salts showed similar solubility in FeSSIF and FaSSIF, but differences were observed in maleate in gastric juice.

[0130] Under these testing conditions, salts and free bases within the biorelevant range are similar in efficacy. However, maleate offers promise when considering intestinal transitions and overall solid form efficacy. Although hydrochlorides are soluble, they are polymorphic and have complex thermal profiles (indicating hydration and solvation). [Stability over two weeks] [ ] [plan] [ ]

[0131] Experiment: Maleate pattern B (30 mg) was placed in individual 15 mL Type I glass vials. HDPE plastic caps were loosely attached to these vials to allow moisture ingress. The vials were then placed in ICH-rated stability chambers at 25°C / 60% RH and 40°C / 75% RH, and in a cold storage at 2–8°C. After two weeks of storage, the samples were removed from the stability chambers and cold storage, and their chemical purity was assessed by HPLC (Method 2). Data were collected at a wavelength of 302 nm. The samples were prepared in MeCN:water (1:1).

[0132] maleate pattern B is stable for two weeks under the following conditions: 25℃ / 60%RH, 40℃ / 75%RH and 2-8℃ Point in time / storage T=0 T = 2 weeks 2-8℃ T = 2 weeks 25℃ / 60%RH T = 2 weeks 40℃ / 75%RH HPLC purity (HPLC method 2) 96.85 96.85 96.99 96.98

[0133] The four optimal salts are maleate, toluenesulfonate, benzenesulfonate, and malonate. Among them, maleate exhibits the best properties. The selected crystalline forms of these salts are described in more detail below. [X] [Powder diffraction] [Research] [ ] [Cibutenedioate pattern] [B]

[0134] The XRPD spectrum of maleate pattern B is shown in Figure 25, and the thermal data is shown in Figure 26. The XRPD peaks of pattern B are described in the table below. Location [°2Th.] Altitude [cts] FWHM [°2Th.] d-spacing[A) Relative strength [%) 5.6284 187.86 0.6140 15.70212 3.50 6.9241 4611.31 0.0768 12.76650 86.00 9.3940 2052.07 0.0768 9.41471 38.27 11.7686 4879.50 0.1023 7.51989 91.00 12.4667 224.33 0.1023 7.10028 4.18 13.1479 452.73 0.1023 6.73392 8.44 13.4712 887.44 0.1023 6.57305 16.55 14.0901 1108.81 0.1023 6.28568 20.68 14.4106 451.78 0.1279 6.14658 8.43 15.6116 2210.87 0.1279 5.67633 41.23 15.8143 2118.49 0.1023 5.60405 39.51 16.1313 475.62 0.1279 5.49462 8.87 16.5029 216.63 0.1023 5.37172 4.04 17.2705 682.34 0.1535 5.13467 12.73 17.6733 2969.71 0.1023 5.01853 55.38 17.9199 4342.69 0.1279 4.95003 80.99 18.2543 1106.15 0.1279 4.86010 20.63 18.6741 1986.10 0.1279 4.75178 37.04 19.1654 995.76 0.1279 4.63105 18.57 19.9459 98.11 0.1535 4.45156 1.83 20.7957 450.57 0.1535 4.27154 8.40 21.8726 1930.42 0.1535 4.06361 36.00 22.3012 1015.57 0.1279 3.98647 18.94 22.7998 134.49 0.1535 3.90041 2.51 23.6624 344.56 0.1279 3.76014 6.43 23.9351 633.64 0.1279 3.71791 11.82 24.8019 214.73 0.1279 3.58991 4.00 25.0411 212.35 0.1023 3.55615 3.96 25.7311 769.93 0.1279 3.46234 14.36 26.1114 649.57 0.0768 3.41276 12.11 26.4273 5361.98 0.1791 3.37269 100.00 26.8310 2088.44 0.1535 3.32284 38.95 27.3094 785.65 0.1535 3.26572 14.65 27.7550 1972.89 0.1791 3.21429 36.79 29.0081 581.05 0.2047 3.07822 10.84 29.6415 151.62 0.1791 3.01387 2.83 30.5909 242.30 0.1791 2.92247 4.52 31.7416 419.84 0.1535 2.81910 7.83 32.3802 138.18 0.2558 2.76495 2.58 33.4454 85.94 0.1535 2.67928 1.60 34.6245 22.98 0.1535 2.59069 0.43 [Citric acid salt pattern] [A]

[0135] The XRPD spectrum of maleate pattern A is shown in Figure 27, and the thermal data is shown in Figure 28. The XRPD peaks of pattern A are described in the table below. Location [°2Th.] Altitude [cts] FWHM [°2Th.] d-Spacing [Å] Relative strength [%) 5.2880 79.08 0.5117 16.71232 8.82 6.5960 896.14 0.1791 13.40077 100.00 9.2386 337.42 0.1535 9.57269 37.65 11.1317 489.61 0.1279 7.94868 54.64 11.5356 253.70 0.1023 7.67125 28.31 14.2833 284.59 0.1535 6.20110 31.76 15.6422 179.45 0.1535 5.66529 20.02 16.0500 171.59 0.1023 5.52227 19.15 16.9462 219.43 0.1535 5.23218 24.49 17.3430 619.49 0.1279 5.11335 69.13 18.5483 272.31 0.2047 4.78371 30.39 18.9422 154.41 0.0553 4.68513 17.23 19.7251 68.05 0.0900 4.50089 7.59 20.5367 212.24 0.1279 4.32482 23.68 21.6425 113.90 0.3070 4.10628 12.71 22.0699 126.37 0.1535 4.02772 14.10 22.8840 84.48 0.2047 3.88625 9.43 25.5206 110.36 0.2047 3.49041 12.31 25.8745 263.80 0.1092 3.44347 29.44 26.5069 385.90 0.1535 3.36274 43.06 27.6811 75.78 0.0900 3.22270 8.46 28.6708 111.36 0.2047 3.11367 12.43 [Cibutenedioate pattern] [C]

[0136] The XRPD spectrum of maleate pattern C is shown in Figure 29, and the thermal data is shown in Figure 30. The XRPD peaks of pattern C are described in the table below. Location [°2Th.] Altitude [cts] FWHM [°2Th.] d-Spacing [Å] Relative strength [%) 5.3327 83.03 0.4093 16.57231 5.52 6.6739 1504.73 0.1791 13.24447 100.00 9.1925 817.35 0.1279 9.62061 54.32 11.1462 422.05 0.1279 7.93834 28.05 11.5411 935.39 0.1535 7.66761 62.16 13.2677 195.85 0.1535 6.67338 13.02 13.8736 293.30 0.1279 6.38327 19.49 14.2687 541.48 0.1791 6.20742 35.99 15.6379 768.88 0.1279 5.66687 51.10 15.9603 359.30 0.1279 5.55308 23.88 16.9613 395.42 0.1791 5.22755 26.28 17.3625 1072.27 0.1279 5.10766 71.26 17.7298 1113.95 0.1279 5.00266 74.03 18.5399 668.08 0.2047 4.78585 44.40 20.5008 285.44 0.2558 4.33231 18.97 21.6905 539.92 0.1535 4.09730 35.88 22.1596 358.58 0.2047 4.01163 23.83 23.7465 154.55 0.1535 3.74701 10.27 25.5692 386.89 0.2047 3.48389 25.71 26.2698 1327.41 0.1535 3.39254 88.22 27.5980 420.43 0.1791 3.23222 27.94 28.8551 190.05 0.6140 3.09420 12.63 30.4932 57.31 0.3070 2.93161 3.81 31.5105 77.47 0.6140 2.83925 5.15 [Maldives Pattern] [B]

[0137] The XRPD spectrum of malonic acid ester pattern B is shown in Figure 31, and the DSC and TGA traces are shown in Figure 32. The XRPD peaks are listed in the table below. Location [°2Th.] Altitude [cts] FWHM [°2Th.] d-Spacing [Å] Relative strength [%) 6.4684 2427.30 0.1023 13.66496 100.00 7.7000 204.63 0.0768 11.48178 8.43 9.3757 142.91 0.1023 9.43303 5.89 10.5956 1899.35 0.1023 8.34958 78.25 11.2972 114.47 0.1023 7.83254 4.72 13.2150 287.40 0.1023 6.69989 11.84 14.2527 666.30 0.1535 6.21434 27.45 14.8453 202.81 0.1023 5.96757 8.36 15.7156 313.92 0.1791 5.63902 12.93 16.5805 729.51 0.1535 5.34677 30.05 17.0621 320.41 0.3048 5.19691 13.20 17.3089 335.35 0.1279 5.12335 13.82 17.9127 194.99 0.1023 4.95199 8.03 18.3739 675.01 0.1279 4.82872 27.81 19.9851 74.62 0.0900 4.44292 3.07 20.4457 290.41 0.1279 4.34387 11.96 20.9222 146.01 0.1535 4.24600 6.02 21.5253 193.94 0.1535 4.12838 7.99 22.5851 43.74 0.0900 3.93699 1.80 22.9598 73.53 0.1535 3.87358 3.03 23.5282 244.44 0.1279 3.78128 10.07 24.6490 93.06 0.1279 3.61183 3.83 25.4768 373.34 0.1279 3.49631 15.38 25.8559 537.48 0.2555 3.44591 22.14 26.4188 372.48 0.1791 3.37375 15.35 26.7811 276.03 0.1023 3.32892 11.37 27.3171 95.19 0.0900 3.26481 3.92 27.9810 90.05 0.3070 3.18884 3.71 28.7910 91.75 0.2558 3.10094 3.78 29.4577 68.42 0.1535 3.03227 2.82 [Toluenesulfonate pattern] [A]

[0138] The XRPD spectrum of toluenesulfonate pattern A is shown in Figure 33, and the TGA and DSC traces are shown in Figure 34. The XRPD peaks are listed in the table below. Location [°2Th.] Altitude [cts] FWHM [°2Th.] d-Spacing [Å] Relative strength [%) 5.3623 94.92 0.5117 16.48094 6.90 7.4588 129.28 0.0768 11.85248 9.39 8.3404 226.46 0.1023 10.60158 16.45 8.8036 572.85 0.0768 10.04469 41.61 9.0769 1376.59 0.1023 9.74292 100.00 9.5099 177.22 0.1023 9.30022 12.87 10.8948 62.90 0.3070 8.12092 4.57 11.6679 660.29 0.1023 7.58452 47.97 13.7713 704.95 0.1023 6.43045 51.21 14.3266 249.95 0.1023 6.18246 18.16 14.8992 743.71 0.1535 5.94609 54.03 15.7056 500.19 0.1279 5.64257 36.34 16.4685 421.65 0.1023 5.38288 30.63 17.8776 425.16 0.1023 4.96165 30.89 18.6033 156.43 0.2047 4.76969 11.36 19.1160 117.00 0.1023 4.64292 8.50 19.6343 107.84 0.1023 4.52152 7.83 20.1841 168.51 0.1279 4.39956 12.24 21.7448 180.09 0.1279 4.08720 13.08 22.2203 854.48 0.1279 4.00080 62.07 22.5619 302.16 0.1279 3.94098 21.95 23.3509 126.53 0.1791 3.80959 9.19 24.1135 221.69 0.1535 3.69081 16.10 24.8389 320.64 0.1279 3.58463 23.29 26.3188 35.80 0.2047 3.38634 2.60 27.3858 77.29 0.2047 3.25678 5.61 27.8900 66.59 0.1535 3.19904 4.84 29.2068 88.90 0.1535 3.05773 6.46 30.2006 39.65 0.2047 2.95934 2.88 32.4628 17.05 0.8187 2.75810 1.24 [Benzenesulfonate pattern] [C]

[0139] The XRPD spectrum of benzenesulfonate pattern C is shown in Figure 35, and the TGA and DSC traces are shown in Figure 36. The XRPD peaks are listed in the table below. Location [°2Th.] Altitude [cts] FWHM [°2Th.] d-interval [Å] Relative strength [%] 5.3099 88.50 0.5117 16.64342 5.28 6.4346 49.98 0.3070 13.73667 2.98 9.4241 614.80 0.1023 9.38471 36.66 11.2303 980.94 0.1023 7.87909 58.49 12.8313 123.05 0.1279 6.89936 7.34 13.3125 865.00 0.1279 6.65103 51.57 14.0143 126.51 0.1023 6.31949 7.54 14.6542 1108.95 0.1279 6.04495 66.12 15.4744 1677.21 0.1279 5.72635 100.00 16.0933 847.34 0.1023 5.50749 50.52 16.2732 561.39 0.0768 5.44701 33.47 18.1250 984.95 0.1023 4.89448 58.73 19.1636 351.15 0.1279 4.63148 20.94 20.3168 109.03 0.1535 4.37113 6.50 20.9164 1037.05 0.1023 4.24716 61.83 21.2514 219.63 0.0768 4.18097 13.10 22.2353 95.89 0.1535 3.99814 5.72 22.8379 195.27 0.1023 3.89399 11.64 23.1023 139.88 0.1535 3.85001 8.34 24.1395 821.07 0.1279 3.68689 48.95 25.4472 1090.62 0.1535 3.50032 65.03 26.0941 280.14 0.1535 3.41498 16.70 26.4366 592.68 0.1535 3.37151 35.34 27.0051 350.71 0.1279 3.30181 20.91 29.3206 118.40 0.1535 3.04613 7.06 29.7647 126.30 0.1279 3.00168 7.53 30.2846 37.92 0.1535 2.95133 2.26 30.8533 82.31 0.1279 2.89821 4.91 32.4258 40.50 0.3582 2.76116 2.41 33.4370 33.42 0.4093 2.67994 1.99 [Gravimetric Gas Phase Adsorption Study] [ ]

[0140] The GVS data obtained using the scheme described above are presented below for certain crystallization forms of the salt. [Cibutenedioate pattern] [A -] [See figure]

[44]

[0141] During the initial adsorption cycle, the solids increased by 1.5 wt% from 50% RH to 90% RH. In the subsequent desorption cycle, the solids lost 4% water, decreasing to 0% RH. In subsequent adsorption cycles, this increased to 4 wt% at 90% RH. GVS curves confirmed that this water absorption system is reversible with decreasing relative humidity, with only a slight hysteresis observed.

[0142] The new pattern was isolated at 0% and 90% RH and named B. Pattern B is closely related to pattern A. [Cibutenedioate pattern] [B -] [See figure]

[45]

[0143] During the initial desorption cycle, the solid lost 2.5 wt% of water from 50% RH to 0% RH. In subsequent adsorption cycles, the solid gained 4% water up to 90% RH, with a sharp increase observed between 0% RH and 40% RH. At 0% RH, it transformed into pattern A, while at 90% RH, there was no change. This data suggests that the interconversion between crystalline forms is related to hydration. [Toluenesulfonate pattern] [A -] [picture] [4]6

[0144] During the initial desorption cycle, the solids lost 3.5 wt% from 50% RH to 0% RH, with a steady decrease of 0.5 wt% from 50% RH to 10% RH, followed by a sharp decrease of approximately 3 wt% from 10% RH to 0% RH. In subsequent adsorption cycles, the solids rapidly increased by approximately 3% water up to 10% RH, and then steadily increased by approximately 1% from 10% RH to 90% RH. A 3% water content is equivalent to the monohydrate of toluenesulfonate. The absence of form changes at 0% RH and 90% RH indicates that the channel hydrate is reversible and stable across the entire environmental range. [Benzenesulfonate pattern] [A -] [See figure]

[47]

[0145] During the initial desorption cycle, the solids lost 2.5 wt% from 50% RH to 0% RH. In subsequent adsorption cycles, the solids gained 10% water up to 90% RH, and increased sharply by 6% from 40% RH to 60% RH. The following desorption cycle shows a steady decrease of approximately 3 wt% from 90% RH to 30% RH, followed by a sharp decrease to 0% wt% from 30% RH to 0% RH. The theoretical water content required for the benzenesulfonate monohydrate form is 3.6%. Therefore, this type of brine is converted to a trihydrate. [Benzenesulfonate pattern] [B -] [See figure]

[48]

[0146] During the initial desorption cycle, the solid lost 1 wt% of water from 50% RH to 0% RH. In subsequent adsorption cycles, the solid gained approximately 2.25% water up to 90% RH, with a noticeable steady increase. Pattern B of the benzenesulfonate shows a corrected GVS curve relative to pattern A. [Benzenesulfonate pattern] [C -] [See figure]

[49]

[0147] During the initial desorption cycle, the solid lost approximately 3 wt% of water from 50% RH to 0% RH. In subsequent adsorption cycles, the solid gained 3.75% water up to 90% RH, with a sharp increase of approximately 2.5 wt% observed between 0% RH and 20% RH. [Naphthalene] [-2-] [Sulfonate pattern] [A -] [See figure]

[50]

[0148] During the initial desorption cycle, the solid lost 1 wt% of water from 50% RH to 0% RH. In subsequent adsorption cycles, the solid gained approximately 2.25% water up to 90% RH, with a noticeable steady increase. XRPD analysis showed no change in the crystallinity of the solid under extreme humidity conditions. [Maldives Pattern] [B -] [See figure]

[51]

[0149] The GVS curves show that the material lost 5 wt% during the initial desorption step up to 0% RH. Therefore, the material is considered hygroscopic and hydrated to non-stoichiometric levels under ambient conditions. In subsequent adsorption cycles, the solid gained 7.5% water up to 90% RH, with a sharp increase of approximately 3 wt% observed between 30% and 40% RH. This hygroscopic system is reversible; the absorbed water is lost as the relative humidity decreases. The theoretical water content required for the malonate monohydrate form is 3.4%, therefore the salt hydrates to dihydrate levels under extreme humidity. [Example] [4] [ ] [Further research on maleic acid salts]

[0150] Five crystal patterns of maleate were identified and labeled as Pattern A, Pattern B, Pattern C, Pattern D, and Pattern E. Characterization data for Patterns A, B, and C are described above, and characterization data for Patterns D and E are described below.

[0151] A comparison of the XRPD spectra of the five crystal patterns with that of a mixture of A / B patterns is shown in Figure 52.

[0152] Patterns A, B, C, and D appear to be variants with varying degrees of hydration. Pattern A has been found to be difficult to separate because it becomes a mixture of A and B once it absorbs any water. Pattern B is a relatively stable hydrate, while pattern C is considered a non-stoichiometric hydrate. Pattern D is also considered a non-stoichiometric hydrate and similar to pattern C. Pattern E is an N-methylpyrrolidone (NMP) solvate. [4A.] [via pattern] [A / B] [Preparation of maleate patterns using mixtures] [A] [,] [Subsequent thermal cycling] [ ]

[0153] A free base of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxynitrile (4.9756 g) was added to a round-bottom flask along with THF (204 mL, 41 volumes). The mixture was heated to 60°C. Then, maleic acid (1.48 g, 1 equivalent) in THF (2 volumes) was added to the solution. The mixture was equilibrated at 60°C for 1 hour, then cooled to 20°C and maintained overnight to obtain a beige suspension. The solid was separated by vacuum filtration and washed with THF. The solid was dried under vacuum at 40°C for 20 hours to obtain maleic acid salt (SSA203).

[0154] A portion of the obtained solid (SSA203) was weighed into a crystallization tube (50 mg) and methyl isobutyl ketone (5 volumes) was added. The mixture was allowed to equilibrate at room temperature for 18 hours to obtain a mixture of pattern A and B. The mixture was then thermally cycled to 150°C to obtain maleate pattern A. [4B.] [Preparation of maleate patterns using high-boiling-point non-aqueous solvents] [A] [ ]

[0155] SSA203 (from Example 4A) was weighed into a crystallization tube (60 mg / tube) and a suitable high-boiling solvent (10 volumes) was added. The mixture was equilibrated at room temperature for about 30 minutes, heated to 95°C and equilibrated for 4 hours, and then allowed to cool naturally to room temperature for 70 hours. The mixture was then heated again to 95°C, equilibrated for 4 hours, and then allowed to cool to room temperature for 3 hours. The solid was separated and dried at 45°C for 18 hours.

[0156] The solvent and the resulting maleate pattern are shown in the table below. [Sample Identification] [solvent] [(] [volume] [)] [XRPD] [Dry solids] [SSA243-A] Xylene (10) Pattern A [SSA243-D] n-PrOAc (10) Pattern A [SSA243-F] Decahydronaphthalene (10) Pattern A [SSA243-G] Dimethyl ether (10) Pattern A [SSA243-H] 1-BuOH (10) Pattern A [4C.] [Antisolvent-mediated recrystallization] [ ]

[0157] Weigh 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxynitrile maleate (from SSA203 in Example 4A) into two crystallization tubes and fill them with DMSO (4 volumes) or NMP (4 volumes). Heat the mixture to 60°C. Then clarify the yellow solution at 60°C into clean, preheated tubes. Then divide the clarified solution into 320 µl aliquots so that each tube contains 80 mg of maleate.

[0158] Next, add appropriate antisolvent in the form of 0.5 to 1 volume portions of the sample to the solution, and equilibrate for at least 10 minutes after each addition until a turbid solution is formed or until 10 volumes of antisolvent are added.

[0159] The mixture was then equilibrated at 60°C for approximately 30 minutes and subsequently cooled to 25°C and equilibrated for approximately 20 hours.

[0160] The other items, which were kept in solution, were cooled to 0°C and equilibrated for approximately 6 hours. The mixture, which was kept in solution at 0°C, was heated to 60°C, and approximately half of the solvent was evaporated by a gentle nitrogen flow and then cooled back to ambient temperature.

[0161] The crystallization patterns obtained from various solvent combinations are shown below. Pattern B of solids separated from DMSO / water, NMP / MeCN and NMP / water Pattern A / B mixture from NMP / BuOH Pattern C separated from DMSO / BuOH and DMSO / MeCN Pattern D of rapid separation from THF+ Pattern E separated from NMP / dioxane, NMP / n-PrOAc, NMP / toluene, NMP / THF, and NMP / EtOAc The DSC and TGA curves of maleate pattern E are shown in Figure 55. [4D.] [Cibutenedioate pattern] [D] [Of] [preparation] [ ]

[0162] The free base (4.9756 g) of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxynitrile was added to a round-bottom flask along with THF (204 mL, 41 volumes). The mixture was heated to 60°C. Then, maleic acid (1.48 g, 1 equivalent) in THF (2 volumes) solution was added to the solution. The mixture was allowed to equilibrate at 60°C for 1 hour. 100 mL of the solution was clarified at 60°C into a clean, preheated flask, cooled to approximately 50°C, and rapidly evaporated to obtain maleic acid salt pattern D.

[0163] The DSC and TGA curves of maleate pattern E are shown in Figure 54. [4E.] Synthesis of Amorphous Cis-Butenedioate [ ]

[0164] 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxynitrile maleate (583.4 mg) was dissolved in hexafluoro-2-propanol (6F-IPA, 6 volumes, 1750 µL) at 30 °C. The solution was clarified into a tube containing tertiary butyl methyl ether (TBME, 6 mL) and cooled to 0 °C. The mixture was stirred at 0 °C for 15 minutes, and the solid was separated by filtration under vacuum and dried at 45 °C over a period of 18 hours. [4F.] [By adjusting the pattern] [A] [Forms maleic anhydride] [B] [ ]

[0165] The maleate pattern A was conditioned for 48 hours using a warm vacuum oven (25°C, slight vacuum degassing to provide active flow through the oven) and a moisture source (static, deionized water tray), and was continuously monitored by a multi-sample method (XRPD sample) through the conditioning tray until all samples reported pattern B. [4G.] [Pattern of maleic anhydride] [B] [Dynamic gas-phase adsorption] (DVS) [analyze] [ ]

[0166] Under ambient conditions, a defined amount of maleate pattern B is placed in a weighing mesh stainless steel basket. A complete experimental cycle consists of five scans (desorption, adsorption repetition, and desorption) at constant temperature (25°C) and 10% RH intervals (60 minutes at each humidity level) within a range of 0-90%. This type of extended experiment should demonstrate the ability of the sample under study to absorb (or not absorb) moisture within a well-defined range of humidity.

[0167] After cycling, the material was separated at 0% RH and the crystallinity was tested, and then kept at 90% RH for at least 3 hours and the change in crystallinity was retested.

[0168] The results are shown in Figure 53.

[0169] The solid exhibited approximately 2.8 wt% associated moisture prior to the first desorption. During the first desorption, the main weight increase was between 20 and 30% RH (approximately 2 wt%). After 5 cycles, the material returned to 0% RH with no associated moisture.

[0170] XRPD analysis indicates the mixed phase at 0% RH and pattern B at 90% RH. This curve, exhibiting a relevant hysteresis between 30% and 0% RH, is typical of reversible channel hydrates, whose kinetic transition from anhydrous to hydrate requires a time above 30% RH to reach equilibrium. Bioactivity [Example] [A] [Chk-1] [Kinetic inhibitory activity] [ ]

[0171] The activity of compound (5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxynitrile) against Chk-1 kinase was tested using the materials and protocols described below. [Reaction buffer] [:] [ ]

[0172] Basic reaction buffer: 20 mM Hepes (pH 7.5), 10 mM MgCl2, 1 mM EGTA, 0.02% Brij35, 0.02 mg / ml BSA, 0.1 mM Na3VO4, 2 mM DTT, 1% DMSO

[0173] *Add the required cofactors individually to each kinase response.

[0174] [Reaction Procedure:] [ ] (i) Prepare the specified substrate in the freshly prepared basal reaction buffer. (ii) Deliver any desired cofactors to the above-mentioned substrate solution. (iii) Deliver the designated kinase into the recipient solution and mix gently. (iv) Delivery of compounds from DMSO into the kinase reaction mixture. (v) 33P-ATP (specific activity final 0.01 μCi / μL) was delivered into the reaction mixture to initiate the reaction (vi) The kinase reactants were incubated for 120 min at room temperature (vii) How to attach the reactants to the P81 ion exchange paper (Whatman No. 3698-915) (viii) Wash the filter thoroughly in 0.1% phosphoric acid (ix) Dry the filter and measure the count in the scintillation counter

[0175] [Kinase Information] [:] [ ] CHK-1 - Genbank deposit number AF016582 Recombinant full-length constructs, N-terminal GST labeled, purified from insect cells. No specific measures were taken to activate this kinase. Final concentration in analysis = 0.5 nM Recipient: CHKtide Peptide sequence: [KKKVSRSGLYRSPSMPENLNRPR] Final concentration in analysis = 20µM No additional cofactors were added to the reaction mixture Based on the results obtained by following the above protocol, the IC50 value of compounds of formula (1) against Chk-1 kinase has been determined to be 0.00015 μM. [instance] [B] [ ] [Cell analysis of gemcitabine combination]

[0176] Exponentially growing MIA PaCa-2 (ATCC CRL-1420) cells were treated with trypsin to remove them from the disk surface. Approximately 10,000 cells / well were seeded in 96-well disks with RPMI containing 10% fetal bovine serum, 1% sodium pyruvate, and 1% L-GlutaMax. Cells were allowed to adhere to the disk surface overnight. Serial semi-logarithmic dilutions of the Chk1 inhibitor test compound and gemcitabine were performed at final maximum concentrations of 3000 nM and 100 nM, respectively. Chk1 inhibitors and gemcitabine were combined to add each concentration of Chk1 inhibitor to each concentration of gemcitabine. Each drug was also tested as a single agent. The drug was added to the adherent cells (in duplicate) and incubated for 72 hours. At 72 hours, the cells were treated with Promega Cell Titer Glo reagent for approximately 15 minutes. Emissions (relative light units, RLU) were recorded using a BMG Polarstar Omega disk reader. The concentration of a single drug that resulted in a 50% reduction in total signal (IC50) was calculated using PRISM software and four-parameter nonlinear regression curve fitting. For combination studies, the relative efficiency (RLU) was plotted on an XY plot using PRISM, with gemcitabine concentration on the X-axis and RLU on the Y-axis. The RLU of each Chk1 inhibitor concentration was plotted as a function of gemcitabine concentration. The IC50 of gemcitabine alone and at each Chk1 concentration was determined using four-parameter nonlinear regression curve fitting. The approximate concentrations of Chk1 inhibitors that resulted in a two-fold and ten-fold reduction in the IC50 of gemcitabine alone were calculated as indicators of synergistic efficacy.

[0177] Based on the results obtained following the above protocol, the approximate compound concentrations for the IC50 values ​​(Chk1 IC50) of compound (1) alone against MIAPaca-2 cells, and for compound (1) to reduce the IC50 of gemcitabine alone by two-fold (2xLS) and ten-fold (10xLS) are shown below. [Chk1 IC, 50 , (nM)] [2xLS (nM)] [10xLS (nM)] 144 3 100 [Pharmaceutical preparations] [(i)] [Tablet formulation]

[0178] The tablet composition containing a pharmaceutically acceptable salt as defined in any of Examples 1.1 to 1.48 or above is prepared by mixing 50 mg of the compound with 197 mg of lactose (BP) as a diluent and 3 mg of magnesium stearate as a lubricant and compressing in a known manner to form a tablet. [(ii)] [Capsule formulation]

[0179] The capsule formulation was prepared by mixing 100 mg of a pharmaceutically acceptable salt as defined in any of Examples 1.1 to 1.48 or above with 100 mg of lactose and filling the resulting mixture into standard opaque hard gelatin capsules. [(iii)] [Injectable compound] [I]

[0180] The non-enteric composition for injection can be prepared by dissolving a pharmaceutically acceptable salt as defined in any of Examples 1.1 to 1.48 or above in water containing 10% propylene glycol to obtain an active compound concentration of 1.5% by weight. The solution is then sterilized by filtration, filled into ampoules, and sealed. [(iv)] [Injectable compound] [II]

[0181] The non-enteral composition for injection is prepared by dissolving a pharmaceutically acceptable salt (2 mg / ml) and mannitol (50 mg / ml) as defined in any of Examples 1.1 to 1.48 or above in water, sterilely filtering the solution and filling it into a sealable 1 ml vial or ampoule. [(v)] [Injectable compound] [III] [ ]

[0182] The preparation for intravenous delivery by injection or infusion can be prepared by dissolving a pharmaceutically acceptable salt as defined in any of Examples 1.1 to 1.48 or above in water at a concentration of 20 mg / ml. The vial is then sealed and sterilized by autoclaving. [(vi)] [Injectable compound] [IV] [ ]

[0183] The preparation for intravenous delivery by injection or infusion can be prepared by dissolving a pharmaceutically acceptable salt as defined in any of Examples 1.1 to 1.48 or above at 20 mg / ml in water containing a buffer (e.g., 0.2 M acetate, pH 4.6). The vial is then sealed and sterilized by autoclaving. [(vii)] [Subcutaneous injection preparation] [ ]

[0184] The composition for subcutaneous administration is prepared by mixing a pharmaceutically acceptable salt as defined in any of Examples 1.1 to 1.48 or above with pharmaceutical-grade corn oil to obtain a concentration of 5 mg / ml. The composition is sterilized and filled into a suitable container. [(viii)] [Freeze-dried formulations] [ ]

[0185] Aliquots of pharmaceutically acceptable salts as defined in any of Examples 1.1 to 1.48 or above were placed in 50 ml vials and lyophilized. During lyophilization, the composition was frozen at -45°C using a one-step freezing protocol. The temperature was raised to -10°C for annealing, then lowered to freeze at -45°C, followed by a first drying at +25°C for approximately 3400 minutes, and then a second drying step with an additional temperature of 50°C. The pressure was set at 80 mTorr during both the first and second drying processes. [Equivalent] [ ]

[0186] The foregoing examples are presented for illustrative purposes and should not be construed as imposing any limitations on the scope of the invention. It will be readily apparent that many modifications and alterations can be made to the specific embodiments of the invention described above and illustrated in the examples without departing from the fundamental principles of the invention. This application is intended to cover all such modifications and alterations.

[0187] <![CDATA[ <110> PharmaEngine, Inc. Sentinel Oncology Limited (UK) <![CDATA[ <120> Pharmaceutical salts <![CDATA[ <130> P41041TW]]> <![CDATA[ <140> TW 111120730]]> <![CDATA[ <141> 2022-06-02 <![CDATA[ <150> GB 2107924.9 <![CDATA[ <151> 2021-06-03 <![CDATA[ <160> 1 ]]> <![CDATA[ <170> PatentIn version 3.5]]> <![CDATA[ <210> 1]]> <![CDATA[ <211> 23]]> <![CDATA[ <212> PRT]]> <![CDATA[ <213> Artificial sequence <![CDATA[ <220> ]]> <![CDATA[ <223> synthetic sequence]]> <![CDATA[ <400> 1]]> Light Light Light Val Ser Arg Ser Gly Leu Tyr Arg Ser Pro Ser Met Pro 1 5 10 15 Glu Asn Leu Asn Arg Pro Arg 20

[0188]

Claims

1. A pharmaceutically acceptable salt of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxynitrile, which is a maleate.

2. The medically acceptable salt, as requested in item 1, has an acid-to-free base ratio of approximately 1:

1.

3. The pharmaceutically acceptable salt of claim 1 is a maleate pattern B salt having an XRPD spectrum (Cu Kα radiation) characterized by a dominant °2Th (°2θ) peak at 6.9 ±0.2º and / or 26.4 ±0.2º and / or 11.8 ±0.2º and / or 17.9 ±0.2º.

4. The pharmaceutically acceptable salt of claim 1, which is a salt of maleate pattern C, having an XRPD spectrum (Cu Kα radiation) characterized by a dominant 2Th peak at 6.7 ±0.2º and / or 9.2 ±0.2º and / or 11.5 ±0.2º and / or 15.6 ±0.2º and / or 17.4 ±0.2º and / or 17.7 ±0.2º and / or 26.3 ±0.2º.

5. A pharmaceutical composition comprising a pharmaceutically acceptable salt and a pharmaceutically acceptable excipient as claimed in any one of claims 1 to 4.

6. A medically acceptable use of a salt as claimed in any of claims 1 to 4, wherein it is used in the manufacture of a medicine for treating cancer.

7. A pharmaceutical combination comprising a pharmaceutically acceptable salt as claimed in any one of claims 1 to 4 and another therapeutically active agent.

8. A method for preparing a pharmaceutically acceptable salt as claimed in any one of claims 1 to 4, the method comprising dispersing 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxynitrile in tetrahydrofuran to form a mixture, heating the mixture to a high temperature in the range of 45°C to 65°C, adding a desired amount of acid to the mixture; maintaining the mixture at or near the high temperature for a defined period of time, and cooling the mixture to separate the pharmaceutically acceptable salt.

9. A method for preparing a pharmaceutically acceptable salt as claimed in any one of claims 1 to 4, the method comprising dispersing 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxynitrile in a mixture of tetrahydrofuran and acetonitrile to form a mixture, heating the mixture to a high temperature in the range of 45°C to 55°C, adding a desired amount of acid to the mixture; maintaining the mixture at or near the high temperature for a defined period of time, and cooling the mixture to separate the pharmaceutically acceptable salt.

10. A method for preparing a pharmaceutically acceptable salt as claimed in any one of claims 1 to 4, the method comprising dispersing 5-[[5-[4-(4-fluoro-1-methyl-4-piperidinyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyridine-2-carboxynitrile in a mixture of tetrahydrofuran and water to form a mixture, heating the mixture to a high temperature in the range of 45°C to 65°C, adding a desired amount of acid to the mixture, maintaining the mixture at or near the high temperature for a defined period of time, and cooling the mixture to separate the pharmaceutically acceptable salt.

11. The method of any one of claims 8 to 10, wherein the acid is maleic acid and the resulting pharmaceutically acceptable salt is a salt of maleic acid pattern A, having an XRPD spectrum (Cu Kα radiation) characterized by a dominant °2Th (°2θ) peak at 6.6 ± 0.2º and / or 17.3 ± 0.2º and / or 11.1 ± 0.2º.

12. The method of claim 11, further comprising converting the maleic acid pattern A salt into a maleic acid pattern B salt by adjusting the maleic acid pattern A salt in an atmosphere with a relative humidity greater than 50%, wherein the maleic acid pattern B salt has an XRPD spectrum (Cu Kα radiation) characterized by a dominant °2Th (°2θ) peak at 6.9 ±0.2º and / or 26.4 ±0.2º and / or 11.8 ±0.2º and / or 17.9 ±0.2º.

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