Pharmaceutical salts of CHK-1 inhibitors

Abstract

The present invention provides pharma- ceutically acceptable salts of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile selected from maleate, tosylate, besylate, and malonate salts, as well as specific crystalline forms of these salts, methods for preparing these salts, pharmaceutical compositions containing these salts, and uses in therapy.

Description

[Technical field]

[0001] The present invention relates to pharmaceutical salts of the Chk-1 inhibitor compound 5-[[5-[4-(4-fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile, processes for their preparation, pharmaceutical compositions containing them, and their use in the treatment of diseases such as cancer. [Background technology]

[0002] Chk-1 is a serine / threonine kinase involved in the induction of cell cycle checkpoints in response to DNA damage and DNA replication stress [Tse ​​et al., Clin. Can. Res. 2007;13(7)]. Cell cycle checkpoints are regulatory pathways that control the order and timing of cell cycle transitions. Many cancer cells have impaired G1 checkpoint activation. 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 abrogates the intra-S-phase and G2 / M checkpoints and has been shown to selectively sensitize tumor cells to 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], hydroxyurea [Nat. Cell. Biol. 2005;7(2):195-20], the nitroimidazole hypoxia targeting agent TH-302 (Meng et al., AACR, 2013 Abstract No. 2389), and ionizing radiation [Clin. Cancer Res. 2010,16,2076]. See also the review article by McNeely et al. [Pharmacology & Therapeutics(2014),142(1):1-10].

[0004] Recently published data also show that Chk-1 inhibitors can act synergistically with 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 proven to be beneficial when combined with immunotherapeutic agents [Mouw et al., Br J Cancer, 2018.(7):933]. Chk-1 inhibitors have been shown to activate cGAS, which induces innate immune responses through STING signaling, and to induce PD-L1 expression and synergize with anti-PD-L1 in vivo [Sen et al., Cancer Discov 2019 (5):646; Sen et al., J Thorac Oncol, 2019.(12):2152].

[0006] Resistance to chemotherapy and radiotherapy, a clinical challenge for conventional treatments, has been linked to activation of the DNA damage response, in which Chk-1 has been implicated [Nature;2006;444(7):756-760;Biochem. Biophys. Res. Commun. 2011;406(1):53-8].

[0007] It is also believed that Chk-1 inhibitors, alone or in combination, may be useful in treating tumor cells in which DNA damage and constitutive activation of checkpoint pathways drive genomic instability, particularly through replication stress. This phenotype is associated with complex karyotypes, for example in samples from acute myeloid leukemia (AML) patients [Cancer Research 2009,89,8652]. Antagonism of Chk-1 kinase in vitro with small molecule inhibitors or RNA interference techniques strongly reduces clonogenicity in AML samples with high levels of DNA damage. In contrast, Chk-1 inhibition has no effect on normal hematopoietic progenitor cells. Furthermore, recent studies have shown that the tumor microenvironment drives genetic instability [Nature;2008;(8):180-192] and that reduction of Chk-1 increases the sensitivity of cells to hypoxia / reoxygenation [Cell Cycle;2010;9(13):2502]. In neuroblastoma, a kinome RNA interference screen demonstrated that reduction of Chk-1 inhibited the growth of eight neuroblastoma cell lines. Tumor cells with Fanconi anemia DNA repair defects showed sensitivity to Chk-1 inhibition [Molecular Cancer 2009,8:24]. The Chk-1 specific inhibitor PF-00477736 inhibited the growth of 30 ovarian cancer cell lines [Bukczynska et al.,23 rdLorne Cancer Conference] and inhibiting the growth of triple-negative breast cancer cells [Cancer Science 2011,102,882]. PF-00477736 also showed selective single-agent activity in mouse models of spontaneous cancer driven by the MYC oncogene [Ferrao et al., Oncogene (August 15, 2011)]. Chk-1 inhibition by RNA interference or selective small molecule inhibitors results in apoptosis of MYC-overexpressing cells in both in vitro and in vivo mouse models of B-cell lymphoma [Hoglund et al., Clinical Cancer Research, 2011]. The latter data suggest that Chk-1 inhibitors may have utility in the treatment of MYC-driven malignancies such as B-cell lymphoma / leukemia, neuroblastoma, and some breast and lung cancers. Chk-1 inhibitors have also been shown to be effective 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 synthetic lethal to the B family of DNA polymerases, increasing replication stress, DNA damage, and cell death [Rogers et al., 2020,80(8);1735]. Other cell cycle regulatory genes have also been reported to confer sensitivity to Chk-1 inhibitors, including CDK2 and POXM1 [Ditano et al., 20201.11(1);7077; Branigan et al., 2021 Cell Reports 34(9):1098808].

[0008] It has also been reported that mutations that reduce the activity of DNA repair pathways can result in synthetic lethal interactions with Chk1 inhibition. For example, mutations that disrupt the RAD50 complex or ATM signaling enhance responsiveness to Chk1 inhibition [Al-Ahmadie et al., Cancer Discov. 2014.(9):1014-21]. Similarly, defects in the Fanconi anemia homologous DNA repair pathway result in sensitivity to Chk1 inhibition [Chen et al., Mol. Cancer 2009 8:24, Duan et al., Frontiers in Oncology 2014 4:368]. Also, human cells with loss of function of the Rad17 gene product are sensitive to Chk1 suppression [Shen et al., Oncotarget, 2015.6(34):35755].

[0009] Various attempts have been made to develop inhibitors of Chk-1 kinase. For example, WO03 / 10444 and WO2005 / 072733 (both in the name of Millennium) disclose aryl / heteroaryl urea compounds as Chk-1 kinase inhibitors. US2005 / 215556 (Abbott) discloses macrocyclic ureas as kinase inhibitors. WO02 / 070494, WO2006014359, and WO2006021002 (both in the name of Icos) disclose aryl and heteroaryl ureas as Chk-1 inhibitors. WO / 2011 / 141716 and WO / 2013 / 072502 both disclose substituted pyrazinyl-phenyl ureas as Chk-1 kinase inhibitors. WO2005 / 009435 (Pfizer) and WO2010 / 077758 (Eli Lilly) disclose aminopyrazoles as Chk-1 kinase inhibitors.

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

[0011] WO2018 / 183891 (Cascadian Therapeutics) discloses a combination of the compound 5-[[5-[4-(4-fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile or a pharma- ceutically acceptable salt thereof with a WEE-1 inhibitor. However, no specific salt of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile is disclosed. Summary of the Invention [Means for solving the problem]

[0012] (The present invention) It has now been found that 5-[[5-[4-(4-fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile forms crystalline salts with several mineral and organic acids. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0013] Thus, the present invention provides in a first aspect (embodiment 1.1) a pharma- ceutically acceptable salt of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile selected from hydrobromide, mesylate, L-tartrate, esylate, L-aspartate, besylate, tosylate, sulfate, phosphate, citrate, acetate, L-glutamate, maleate, gentisate, glucuronate, malonate, naphthylene-2-sulfonate, ethane-1,2-disulfonate, naphthalene-1,5-disulfonate, and oxalate.

[0014] As used herein, the terms "hydrobromide, mesylate, L-tartrate, esylate, L-aspartate, besylate, tosylate, sulfate, phosphate, citrate, acetate, L-glutamate, maleate, gentisate, glucuronate, malonate, naphthylene-2-sulfonate, and oxalate" are used in their conventional sense to refer to salts of hydrobromide, 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, gentisic acid, glucuronate, malonic acid, naphthylene-2-sulfonic acid, ethane-1,2-disulfonic acid, naphthalene-1,5-disulfonic acid, and oxalic acid, respectively.

[0015] Also, some salts of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile have been found to have improved properties compared to the hydrochloride salt disclosed in WO2015 / 12039.

[0016] Thus, in another embodiment (embodiment 1.2), the present invention provides a pharma- ceutically acceptable salt of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile selected from maleate, tosylate, besylate, and malonate.

[0017] The compound 5-[[5-[4-(4-fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile has the following formula (1), and for convenience in this specification, the maleate, tosylate, besylate, and malonate salts of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile may be referred to as the salt of the compound of formula (1) or the salt of the present invention.

[0018] [ka]

[0019] The compound of formula (1) has several basic nitrogen atoms, and in principle can form salts with different salt ratios (i.e., the molar ratio of free base:acid).For example, when the acid is monobasic, monosalts (i.e., when the molar ratio of acid to free base is 1:1) or bis-salts (when the molar ratio of acid to free base is about 2:1) can be prepared depending on the number of molar equivalents of acid used in the method used to form the salt.When a dibasic acid (e.g., dicarboxylic acid) is used to form the salt, hemi-salts (when the molar ratio of acid to base in the salt is 0.5:1), mono-salts, and bis-salts can be formed depending on the specific salt formation conditions employed.

[0020] Thus, in further embodiments (embodiment 1.3 to embodiment 1.9), the present invention provides:

[0021] 1.3 A pharma- ceutically acceptable salt of the maleate salt, 5-[[5-[4-(4-fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile.

[0022] 1.3A A crystalline maleate salt of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile having crystal pattern B as defined herein.

[0023] 1.4 A pharma- ceutically acceptable salt of the tosylate salt, 5-[[5-[4-(4-fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile.

[0024] 1.5 A pharma- ceutically acceptable salt of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile, the besylate salt.

[0025] 1.6 Pharmaceutically acceptable salts of the malonate salt, 5-[[5-[4-(4-fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile.

[0026] 1.7 A pharma- ceutically acceptable salt according to any one of embodiments 1.1 to 1.6, wherein the salt ratio (molar ratio of acid:free base) is about 1:1.

[0027] 1.8 The pharma- ceutically acceptable salt of embodiment 1.3 or embodiment 1.6, wherein the salt ratio (molar ratio of acid:free base) is about 0.5:1.

[0028] 1.9 A pharma- ceutically acceptable salt according to embodiment 1.3 or embodiment 1.5, wherein the salt ratio (molar ratio of acid:free base) is about 2:1.

[0029] The salts of the compounds of formula (1) may be amorphous or substantially crystalline.

[0030] The term "substantially crystalline" refers to a salt having a crystallinity of 50% to 100%. Within this range, the salt may have at least 55% crystallinity, or at least 60% crystallinity, or at least 70% crystallinity, or at least 80% crystallinity, or at least 90% crystallinity, or at least 95% crystallinity, or at least 98% crystallinity, or at least 99% crystallinity, or at least 99.5% crystallinity, or at least 99.9% crystallinity, for example 100% crystallinity.

[0031] Certain salts of the present invention may exist in several different crystalline forms or polymorphs.

[0032] In UK patent application No. 2107924.9, filed June 3, 2021, from which this application claims priority, certain maleate salt forms were designated Pattern A, Pattern A', and Pattern A''. In the present application, these forms have been redesignated Pattern A, Pattern B, and Pattern C, respectively.

[0033] The crystalline form of the salt of the compound of formula (1) preferably has a crystalline purity of at least 90%, more preferably at least 95%, i.e. at least 90% (more preferably at least 95%) of the salt consists of a single crystalline form.

[0034] The crystalline forms of the salts of the invention may be solvated (eg hydrated) or non-solvated (eg anhydrous).

[0035] The term "anhydrous" as used herein does not exclude the possibility that some water may be present on or in the crystalline form of the salt. For example, some water may be present on the surface of the crystalline form of the salt, and a small amount of water may be present inside the crystalline body of the salt. Typically, the anhydrous form contains less than 0.4 water molecules per molecule of the compound of formula (1), and more preferably contains less than 0.1 water molecules, e.g., 0 water molecules, per molecule of the compound of formula (1).

[0036] When crystalline forms are hydrated, they may contain, for example, up to 3 molecules of crystal water, more commonly up to 2 molecules of water, for example, 1 molecule of water or 2 molecules of water. Non-stoichiometric hydrates may be formed, in which the number of water molecules present is less than 1 or is a non-integer. For example, when there is less than 1 molecule of water, there may be, for example, 0.4 molecules, or 0.5 molecules, or 0.6 molecules, or 0.7 molecules, or 0.8 molecules, or 0.9 molecules of water per molecule of Compound (1).

[0037] Therefore, in further embodiments of the present invention (embodiment 1.10 to embodiment 1.11), the following is provided:

[0038] 1.10 The pharma- ceutically acceptable salt according to any one of embodiments 1.1 to 1.9, having a degree of crystallinity of from 50% to 100%.

[0039] 1.11(b) A pharma- ceutically acceptable salt according to embodiment 1.10, having a crystallinity of at least 55%.

[0040] 1.11A The pharma- ceutically acceptable salt according to embodiment 1.11, having a crystallinity of at least 60%.

[0041] 1.11B The pharma- ceutically acceptable salt of embodiment 1.11, having a crystallinity of at least 70%.

[0042] 1.11C The pharma- ceutically acceptable salt of embodiment 1.11, having a crystallinity of at least 80%.

[0043] 1.11D The pharma- ceutically acceptable salt according to embodiment 1.11, having a crystallinity of at least 90%.

[0044] 1.11E The pharma- ceutically acceptable salt of embodiment 1.11, having a crystallinity of at least 95%.

[0045] 1.11F The pharma- ceutically acceptable salt according to embodiment 1.11, having a crystallinity of at least 98%.

[0046] 1.11G The pharma- ceutically acceptable salt according to embodiment 1.11, having a crystallinity of at least 99%.

[0047] 1.11H The pharma- ceutically acceptable salt of embodiment 1.11, having a crystallinity of at least 99.5%.

[0048] 1.11I The pharma- ceutically acceptable salt according to embodiment 1.11, having a crystallinity of at least 99.9%.

[0049] 1.11J The pharma- ceutically acceptable salt according to embodiment 1.11, wherein the crystallinity is 100%.

[0050] Several techniques can be used to characterize crystalline forms, including X-ray powder diffraction (XRPD), single crystal X-ray diffraction, differential scanning calorimetry (DSC), and thermogravimetric analysis (TGA). Gravimetric vapor sorption studies (GVS), such as dynamic vapor sorption (DVS), can characterize the behavior of crystals under various humidity conditions.

[0051] The crystalline structure of a compound can be characterized by X-ray powder diffraction (XRPD), a solid-state technique. XRPD can be performed according to conventional methods such as those described herein (see Examples below) and in "Introduction to X-ray Powder Diffraction," Ron Jenkins and Robert L. Snyder (John Wiley & Sons, Inc., New York, 1996). The presence of defined peaks (as opposed to random background noise) in an XRPD diffractogram indicates that the compound has some degree of crystallinity.

[0052] The powder X-ray pattern of a compound is characterized by the diffraction angle (2θ) (also referred to herein as °2Th or °2Theta) and interplanar spacing (d) parameters of the X-ray diffraction spectrum, which are related by the Bragg equation nλ=2dSinθ, where n=1, λ=wavelength of the X-ray radiation, d=interplanar spacing, and θ=diffraction angle.

[0053] Thus, in further embodiments (embodiment 1.12 to embodiment 1.42), the present invention provides:

[0054] 1.12 A pharma- ceutically acceptable salt of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile as described in embodiment 1.1 having an XRPD spectrum substantially similar to that shown in any one of Figures 5-25, 27, 29, 31, 33, and 35 (disregarding the XRPD spectrum of the free base or amorphous salt form).

[0055] 1.13 A pharma- ceutically acceptable maleic acid Pattern B salt of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile as described in any one of Embodiments 1.2, 1.3, and 1.3A, having an XRPD spectrum substantially similar to that shown in Figure 25.

[0056] 1.14 A pharma- ceutically acceptable maleic acid Pattern B salt of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile as defined in any one of Embodiments 1.2, 1.3, 1.3A, 1.12, and 1.13, having an XRPD spectrum characterized by a major °2Th (°2Theta) peak at 26.3±0.2° (e.g., at a relative intensity of 100%).

[0057] 1.14A A pharma- ceutically acceptable maleic acid Pattern B salt of 5-[[5-[4-(4-Fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile as defined in any one of embodiments 1.2, 1.3, 1.3A, 1.12, and 1.13, having an XRPD spectrum characterized by major °2Th (°2Theta) peaks at 6.9±0.2° and / or 26.4±0.2° and / or 11.8±0.2° and / or 17.9±0.2°.

[0058] 1.15 A pharma- ceutically acceptable maleic acid Pattern B salt according to embodiment 1.14, having an XRPD spectrum characterized by major °2Th peaks at 6.9±0.2°, 26.4±0.2°, 11.8±0.2°, and 17.9±0.2°.

[0059] 1.16 A pharma- ceutically acceptable maleic acid Pattern B salt according to embodiment 1.14 or embodiment 1.15, having an XRPD spectrum characterized by intermediate °2Th peaks at 15.6±0.2° and / or 9.4±0.2° and / or 15.8±0.2° and / or 17.7±0.2° and / or 26.8±0.2°.

[0060] 1.17 A Pharmaceutically acceptable maleic acid Pattern B salt according to embodiment 1.16, having an XRPD spectrum characterized by intermediate °2Th peaks at 15.6±0.2°, 9.4±0.2°, 15.8±0.2°, 17.7±0.2°, and 26.8±0.2°.

[0061] 1.18 A pharma- ceutically acceptable maleic acid Pattern A salt of 5-[[5-[4-(4-Fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile as described in embodiment 1.2, having an XRPD spectrum substantially similar to that shown in Figure 27.

[0062] 1.19 A pharma- ceutically acceptable maleic acid Pattern A salt of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile as defined in embodiment 1.2 or embodiment 1.18 having an XRPD spectrum characterized by major °2Th peaks at 6.6±0.2° and / or 17.3±0.2° and / or 11.1±0.2°.

[0063] 1.20 A pharma- ceutically acceptable maleic acid Pattern A salt of 5-[[5-[4-(4-Fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile as described in embodiment 1.19, having an XRPD spectrum characterized by major °2Th peaks at 6.6±0.2°, 17.3±0.2°, and 11.1±0.2°.

[0064] 1.21 A pharma- ceutically acceptable maleic acid Pattern A salt according to embodiment 1.19 or embodiment 1.20, having an XRPD spectrum characterized by an intermediate °2Th peak at 26.5±0.2° and / or 9.2±0.2° and / or 14.3±0.2° and / or 18.5±0.2° and / or 25.9±0.2° and / or 11.5±0.2° and / or 16.9±0.2° and / or 20.5±0.2° and / or 15.6±0.2°.

[0065] 1.22 A pharma- ceutically acceptable maleic acid Pattern A salt according to embodiment 1.21 having an XRPD spectrum characterized by intermediate °2Th peaks at 26.5±0.2°, 9.2±0.2°, 14.3±0.2°, 18.5±0.2°, 25.9±0.2°, 11.5±0.2°, 16.9±0.2°, 20.5±0.2°, and 15.6±0.2°.

[0066] 1.23 A pharma- ceutically acceptable maleic acid Pattern C salt of 5-[[5-[4-(4-Fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile as described in embodiment 1.2, having an XRPD spectrum substantially similar to that shown in Figure 29.

[0067] 1.24 A pharma- ceutically acceptable maleic acid Pattern C salt of 5-[[5-[4-(4-Fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile as defined in embodiment 1.2 or embodiment 1.23 having an XRPD spectrum characterized by a major °2Th peak at 6.7±0.2° and / or 9.2±0.2° and / or 11.5±0.2° and / or 15.6±0.2° and / or 17.4±0.2° and / or 17.7±0.2° and / or 26.3±0.2°.

[0068] 1.25 A pharma- ceutically acceptable maleic acid Pattern C salt of 5-[[5-[4-(4-Fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile as described in embodiment 1.24 having an XRPD spectrum characterized by major °2Th peaks at 6.7±0.2°, 9.2±0.2°, 11.5±0.2°, 15.6±0.2°, 17.4±0.2°, 17.7±0.2°, and 26.3±0.2°.

[0069] 1.26 A pharma- ceutically acceptable maleic acid Pattern C salt according to embodiment 1.24 or embodiment 1.25, having an XRPD spectrum characterized by an intermediate °2Th peak at 18.5±0.2° and / or 14.3±0.2° and / or 21.7±0.2° and / or 11.1±0.2° and / or 27.6±0.2° and / or 17.0±0.2° and / or 25.6±0.2° and / or 16.0±0.2° and / or 22.2±0.2°.

[0070] 1.27 A pharma- ceutically acceptable maleic acid Pattern C salt according to embodiment 1.26, having an XRPD spectrum characterized by intermediate °2Th peaks at 18.5±0.2°, 14.3±0.2°, 21.7±0.2°, 11.1±0.2°, 27.6±0.2°, 17.0±0.2°, 25.6±0.2°, 16.0±0.2°, and 22.2±0.2°.

[0071] 1.28 A pharma- ceutically acceptable malonic acid Pattern B salt of 5-[[5-[4-(4-Fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile as described in embodiment 1.12, having an XRPD spectrum substantially similar to that shown in FIG.

[0072] 1.29 A pharma- ceutically acceptable malonic acid Pattern B salt of 5-[[5-[4-(4-Fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile as described in embodiment 1.2 or embodiment 1.28, having an XRPD spectrum characterized by a major °2Th peak at 10.6±0.2° and / or 6.5±0.2°.

[0073] 1.30 A pharma- ceutically acceptable malonic acid Pattern B salt of 5-[[5-[4-(4-Fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile as described in embodiment 1.29, having an XRPD spectrum characterized by major °2Th peaks at 10.6±0.2° and 6.5±0.2°.

[0074] 1.31 A pharma- ceutically acceptable malonic acid Pattern B salt of 5-[[5-[4-(4-Fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile as defined in embodiment 1.29 or embodiment 1.30 having an XRPD spectrum characterized by intermediate °2Th peaks at 16.6±0.2° and / or 18.4±0.2° and / or 14.3±0.2° and / or 25.9±0.2°.

[0075] 1.32 A pharma- ceutically acceptable malonic acid Pattern B salt of 5-[[5-[4-(4-Fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile as described in embodiment 1.31 having an XRPD spectrum characterized by intermediate °2Th peaks at 16.6±0.2°, 18.4±0.2°, 14.3±0.2°, and 25.9±0.2°.

[0076] 1.33 A pharma- ceutically acceptable tosylate Pattern A salt of 5-[[5-[4-(4-Fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile as described in embodiment 1.2, having an XRPD spectrum substantially similar to that shown in FIG.

[0077] 1.34 A pharma- ceutically acceptable tosylate Pattern A salt of 5-[[5-[4-(4-Fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile as defined in embodiment 1.2 or embodiment 1.33 having an XRPD spectrum characterized by major °2Th peaks at 9.1±0.2° and / or 22.2±0.2° and / or 14.9±0.2° and / or 13.8±0.2°.

[0078] 1.35 A pharma- ceutically acceptable tosylate Pattern A salt of 5-[[5-[4-(4-Fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile as described in embodiment 1.34, having a spectrum characterized by major °2Th peaks at 9.1±0.2°, 22.2±0.2°, 14.9±0.2° and 13.8±0.2°.

[0079] 1.36 A pharma- ceutically acceptable tosylate Pattern A salt of 5-[[5-[4-(4-Fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile as defined in embodiment 1.34 or embodiment 1.35, having an XRPD spectrum characterized by an intermediate °2Th peak at 11.7±0.2° and / or 8.8±0.2° and / or 15.7±0.2° and / or 17.9±0.2° and / or 16.5±0.2° and / or 24.8±0.2° and / or 22.6±0.2°.

[0080] 1.37 A pharma- ceutically acceptable tosylate Pattern A salt of 5-[[5-[4-(4-Fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile as described in embodiment 1.36, having an XRPD spectrum characterized by intermediate °2Th peaks at 11.7±0.2°, 8.8±0.2°, 15.7±0.2°, 17.9±0.2°, 16.5±0.2°, 24.8±0.2°, and 22.6±0.2°.

[0081] 1.38 A pharma- ceutically acceptable besylate pattern C salt of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile as described in embodiment 1.2, having an XRPD spectrum substantially similar to that shown in FIG.

[0082] 1.39 A pharma- ceutically acceptable besylate pattern C salt of 5-[[5-[4-(4-Fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile as defined in embodiment 1.2 or embodiment 1.38 having an XRPD spectrum characterized by a major °2Th peak at 15.5±0.2° and / or 14.7±0.2° and / or 25.4±0.2° and / or 20.9±0.2° and / or 18.1±0.2° and / or 11.2±0.2° and / or 13.3±0.2° and / or 16.1±0.2°.

[0083] 1.40 A pharma- ceutically acceptable besylate pattern C salt of 5-[[5-[4-(4-Fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile as described in embodiment 1.39 having an XRPD spectrum characterized by major °2Th peaks at 15.5±0.2°, 14.7±0.2°, 25.4±0.2°, 20.9±0.2°, 18.1±0.2°, 11.2±0.2°, 13.3±0.2°, and 16.1±0.2°.

[0084] 1.41 A pharma- ceutically acceptable besylate pattern C salt of 5-[[5-[4-(4-Fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile as defined in embodiment 1.39 or embodiment 1.40 having an XRPD spectrum characterized by an intermediate °2Th peak at 24.1±0.2° and / or 9.4±0.2° and / or 26.4±0.2° and / or 16.3±0.2° and / or 19.2±0.2° and / or 27.0±0.2°.

[0085] 1.42 A pharma- ceutically acceptable besylate pattern C salt of 5-[[5-[4-(4-Fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile as described in embodiment 1.41 having an XRPD spectrum characterized by intermediate °2Th peaks at 24.1±0.2°, 9.4±0.2°, 26.4±0.2°, 16.3±0.2°, 19.2±0.2°, and 27.0±0.2°.

[0086] In the above embodiments, "major °2Th peak" means a peak whose relative intensity (relative to the maximum peak) is at least 50%, and "intermediate peak" means a peak whose relative intensity is 20% to 50%. Peak positions were measured to at least four decimal places, but are given to one decimal place and ±0.2°. Peak positions are listed in descending order of approximately relative intensity.

[0087] The salts of the invention can also be characterized by their thermal behavior, in particular by DSC and TGA analysis. Thus, in a further embodiment, the invention provides:

[0088] 1.43 A pharma- ceutically acceptable maleic acid Pattern B salt of 5-[[5-[4-(4-Fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile as defined in any one of embodiments 1.13 to 1.17, having DSC and TGA characteristics substantially similar to those shown in FIG. 26.

[0089] 1.44 A pharma- ceutically acceptable maleic acid Pattern A salt of 5-[[5-[4-(4-Fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile as defined in any one of embodiments 1.18 to 1.22, having DSC and TGA characteristics substantially similar to those shown in FIG. 28.

[0090] 1.45 A pharma- ceutically acceptable maleic acid Pattern C salt of 5-[[5-[4-(4-Fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile as defined in any one of embodiments 1.23 to 1.27, having DSC and TGA characteristics substantially similar to those shown in FIG. 30.

[0091] 1.46 A pharma- ceutically acceptable malonic acid Pattern B salt of 5-[[5-[4-(4-Fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile as defined in any one of embodiments 1.28 to 1.32, having DSC and TGA characteristics substantially similar to those shown in FIG. 32.

[0092] 1.47 A pharma- ceutically acceptable tosylate Pattern A salt of 5-[[5-[4-(4-Fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile as defined in any one of embodiments 1.33 to 1.37, having DSC and TGA characteristics substantially similar to those shown in FIG. 34.

[0093] 1.48 A pharma- ceutically acceptable besylate Pattern C salt of 5-[[5-[4-(4-Fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile as de?ned in any one of embodiments 1.38 to 1.42, having DSC and TGA properties substantially similar to those shown in FIG.

[0094] Among the various salts of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile as defined and described above and elsewhere in this specification, the maleate salt is a preferred salt.

[0095] The advantage of the maleate salt is that it is crystalline with one thermodynamically favored stable form (pattern B) and has a low propensity for polymorphism.

[0096] Stability studies performed over a two-week period at 25°C / 60% RH and 40°C / 75% RH showed that the maleate salt remained a free-flowing solid with no evidence of deliquescence or aggregation, and no change in polymorphic form, indicating good chemical stability. Data collected over a subsequent six-month period confirmed these initial findings.

[0097] The maleate salt has improved solubility in water compared to the free base, and when evaluated for solubility in biorelevant solvents, it also had improved solubility in gastric juice.

[0098] (Isotopes) The salts defined in any one of embodiments 1.1 to 1.48 may include one or more isotopic substitutions, and a reference to a particular element includes within its scope all isotopes of that element. For example, a reference to hydrogen includes within its scope all isotopes of that element. 1 H, 2 H(D), and 3Similarly, references to carbon and oxygen include those within their scope. 12 C. 13 C, and 14 C, or 16 O and 18 Each of these includes O.

[0099] The isotope may be radioactive or non-radioactive. In one embodiment of the present invention, the salt does not contain a radioactive isotope. Such compounds are suitable for use in therapy. However, in another embodiment, the salt may contain one or more radioactive isotopes. Such salts containing radioactive isotopes may be useful in diagnostic contexts.

[0100] (Method of Preparing the Salt of the Present Invention) The pharma- ceutically acceptable salts of the invention can be prepared from the free base of the compound 5-[[5-[4-(4-fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile (a compound of formula (1)) by the methods described in the Examples below. The compound of formula (1) can be prepared by the methods described in Example 64, Method L of International Patent Application No. WO2015 / 20390, as shown in Reaction Scheme 1 below.

[0101] [ka]

[0102] In further embodiments (embodiment 2.1 to embodiment 2.10), the present invention provides methods of forming a pharma- ceutically acceptable salt of a compound of formula (1) as follows.

[0103] 2.1 A process for preparing a pharma- ceutically acceptable salt as defined in embodiment 1.1 or embodiment 1.2, comprising dispersing 5-[[5-[4-(4-fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile in tetrahydrofuran to form a mixture; heating the mixture to an elevated temperature in the range of 45°C to 65°C, e.g., 55°C to 65°C, in particular about 60°C; adding the requisite amount of acid to the mixture; maintaining the mixture at or near the elevated temperature for a defined period of time; and cooling the mixture to isolate the pharma- ceutically acceptable salt.

[0104] 2.2 The method of embodiment 2.1, wherein the acid is selected from maleic acid, p-toluenesulfonic acid, benzenesulfonic acid, and malonic acid.

[0105] 2.3 A process for preparing a pharma- ceutically acceptable salt as defined in embodiment 1.1 or embodiment 1.2, comprising dispersing 5-[[5-[4-(4-fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile in a mixture of tetrahydrofuran and acetonitrile (e.g., a 1:1 mixture) to form a mixture, heating the mixture to an elevated temperature in the range of 45°C to 55°C (e.g., about 50°C), adding the requisite amount of acid to the mixture, maintaining the mixture at or near the elevated temperature for a defined period of time, and cooling the mixture to isolate the pharma- ceutically acceptable salt.

[0106] 2.4 The method of embodiment 2.3, wherein the acid is selected from maleic acid, p-toluenesulfonic acid, and benzenesulfonic acid.

[0107] 2.5 A method for preparing a pharma- ceutically acceptable salt as defined in embodiment 1.1 or embodiment 1.2, comprising dispersing 5-[[5-[4-(4-fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile in a mixture of tetrahydrofuran and water (e.g., the mixture comprises 75% to 97% (v / v) tetrahydrofuran and 3% to 25% (v / v) water, more preferably about 95% (v / v) tetrahydrofuran and about 5% (v / v) water) to form a mixture; heating the mixture to an elevated temperature in the range of 45°C to 65°C (e.g., about 50°C to 60°C); adding the requisite amount of acid to the mixture; maintaining the mixture at or near the elevated temperature for a defined period of time; and cooling the mixture to isolate the pharma- ceutically acceptable salt.

[0108] 2.6 The method of embodiment 2.5, wherein the acid is selected from maleic acid, p-toluenesulfonic acid, benzenesulfonic acid, and malonic acid.

[0109] 2.7 The method of embodiment 2.1, wherein the required amount of acid is an excess of acid (e.g., up to a molar excess).

[0110] 2.8 The method of embodiment 2.7, wherein the acid is p-toluenesulfonic acid.

[0111] 2.9 The method of embodiment 2.5, wherein the required amount of acid is an excess of acid (e.g., up to a molar excess).

[0112] 2.10 The method of embodiment 2.9, wherein the acid is selected from p-toluenesulfonic acid and benzenesulfonic acid.

[0113] 2.11 The method of any one of embodiments 2.1, 2.3, and 2.5, wherein the acid is maleic acid and the resulting pharma- ceutically acceptable salt is a maleate salt.

[0114] 2.12 The method of embodiment 2.11, wherein said maleate salt is a maleic acid Pattern A salt.

[0115] 2.13 The method of embodiment 2.13, further comprising converting the Pattern A maleate to a Pattern B maleate by conditioning in an atmosphere having a relative humidity of more than 50% (e.g., a relative humidity of 51% to 90% or a relative humidity of 51% to 85%).

[0116] 2.14 The method of embodiment 2.13, wherein the Pattern A maleate is conditioned in an atmosphere having a relative humidity greater than 60% and a temperature in the range of 35-45° C.

[0117] 2.15 The method of embodiment 2.13 or embodiment 2.14, wherein the Pattern A maleate is conditioned in an atmosphere having a relative humidity of 70% to 80%.

[0118] 2.16 The method of embodiment 2.14, wherein the Pattern A maleate is conditioned in an atmosphere having a relative humidity of about 75% and a temperature of about 40° C.

[0119] The particular set of conditions for carrying out the above method are set forth in the Examples below.

[0120] Biological Properties and Therapeutic Uses The compounds of formula (1) and their salts are potent inhibitors of Chk-1 and are therefore expected to be beneficial in the treatment of a wide range of proliferative diseases, either alone or in combination with a variety of chemotherapeutic agents, immunotherapeutic agents, or radiation.

[0121] Thus, in further embodiments (embodiment 3.1 to embodiment 3.10), the present invention provides:

[0122] 3.1 A pharma- ceutically acceptable salt as defined in any one of embodiments 1.1 to 1.48 for use in medicine or therapy.

[0123] 3.2 A pharma- ceutically acceptable salt as defined in any one of embodiments 1.1 to 1.48 for use as a Chk-1 kinase inhibitor.

[0124] 3.3 A pharma- ceutically acceptable salt as defined in any one of embodiments 1.1 to 1.48 for use in enhancing the therapeutic effect of radiotherapy or chemotherapy or immunotherapy in the treatment of a proliferative disease such as cancer.

[0125] 3.4 A pharma- ceutically acceptable salt as defined in any one of embodiments 1.1 to 1.48 for use in the treatment of a proliferative disease, such as cancer.

[0126] 3.5 Use of a pharma- ceutically acceptable salt as defined in any one of embodiments 1.1 to 1.48 for the manufacture of a medicament for enhancing the therapeutic effect of radiotherapy or chemotherapy or immunotherapy in the treatment of a proliferative disease, such as cancer.

[0127] 3.6 The use of a pharma- ceutically acceptable salt as defined in any one of embodiments 1.1 to 1.48 for the manufacture of a medicament for the treatment of a proliferative disease, such as cancer.

[0128] 3.7 A method for the prevention or treatment of a proliferative disease, such as cancer, comprising administering to a patient a pharma- ceutically acceptable salt as defined in any one of embodiments 1.1 to 1.48 in combination with radiation therapy, immunotherapy, or chemotherapy.

[0129] 3.8 A method for the prevention or treatment of a proliferative disease, such as cancer, comprising administering to a patient a pharma- ceutically acceptable salt as defined in any one of embodiments 1.1 to 1.48.

[0130] 3.9 The cancer is a carcinoma, e.g., carcinoma of the bladder, brain, breast, colon, kidney, epidermis, liver, lung, esophagus, gallbladder, ovary, pancreas, stomach, cervix, thyroid, prostate, gastrointestinal system, or skin; a hematopoietic malignancy, such as leukemia, B-cell lymphoma, T-cell lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, hairy cell lymphoma, mantle cell lymphoma, or Burkitt's lymphoma; a hematopoietic malignancy of the myeloid system, e.g., acute myeloid leukemia, chronic myeloid leukemia, myelodysplastic syndrome, or promyeloma. A pharmaceutically acceptable salt, use, or method for use as defined in any one of embodiments 3.3 to 3.8, wherein the tumor is selected from: chronic leukemia; follicular thyroid carcinoma; tumors of mesenchymal origin, such as fibrosarcoma or rhabdomyosarcoma; tumors of the central or peripheral nervous system, such as astrocytoma, neuroblastoma, glioma, medulloblastoma, or neurilemmoma; melanoma; seminoma; teratocarcinoma; osteosarcoma; xeroderma pigmentosum; keratoacanthoma; follicular thyroid carcinoma; Ewing's sarcoma or Kaposi's sarcoma.

[0131] 3.10 A pharma- ceutically acceptable salt, use, or method for use according to embodiment 3.9, wherein said cancer is selected from breast cancer, colon cancer, lung cancer, ovarian cancer, pancreatic cancer, prostate cancer, glioma, Ewing's sarcoma, lymphoma (e.g., mantle cell lymphoma), medulloblastoma, and leukemia.

[0132] It is also believed that the pharma- ceutically acceptable salts of formula (1) described herein may be useful in the treatment of: (a) Cancers driven by oncogenes including Myc and CCNE1; (b) cancers with deregulated cell cycle or DNA damage repair pathways, such as cancers with defects in RAD17 (e.g., RAD17-mutated tumors), RAD50, TP53, or ATM (e.g., tumors with defective DNA repair mechanisms or cell cycle defects, such as cancers with abrogated G1 / S-phase DNA damage checkpoints due to mutations (e.g., p53 mutations)), or Fanconi anemia; and (c) Cancers with high levels of replication stress, such as Chk1 or ATR amplification.

[0133] Thus, in further embodiments (embodiment 3.11 to embodiment 3.23), the present invention provides:

[0134] 3.11 A pharma- ceutically acceptable salt, use, or method for use as defined in any one of embodiments 3.3 to 3.10, wherein said cancer is characterized by defects in DNA repair mechanisms, cell cycle defects, or high levels of replication stress.

[0135] 3.12 A pharma- ceutically acceptable salt, use, or method for use according to embodiment 3.11, wherein said cancer is a p53-negative or p53-mutated tumor.

[0136] 3.13 A pharma- ceutically acceptable salt, use, or method for use as defined in any one of embodiments 3.3 to 3.10, wherein said cancer is a cancer driven by the MYC oncogene.

[0137] 3.14 A pharma- ceutically acceptable salt, use, or method for use according to embodiment 3.13, wherein said cancer driven by the MYC oncogene is B-cell lymphoma, leukemia, neuroblastoma, medulloblastoma, breast cancer, or lung cancer.

[0138] 3.15 A pharma- ceutically acceptable salt as defined in any one of embodiments 1.1 to 1.48 for use in combination with radiation therapy, immunotherapy, or chemotherapy in the treatment of patients with p53-negative or p53-mutated tumors (e.g., a cancer selected from breast cancer, colon cancer, lung cancer, ovarian cancer, pancreatic cancer, prostate cancer, glioma, and leukemia).

[0139] 3.16 A pharma- ceutically acceptable salt for use according to any one of embodiments 3.3 to 3.15, wherein said treatment, in addition to the administration of a pharma- ceutically acceptable salt as defined in any one of embodiments 1.1 to 1.48, comprises administration to the patient of a chemotherapeutic agent selected from cytarabine, etoposide, gemcitabine, cyclophosphamide, a Wee1 inhibitor, and SN-38.

[0140] 3.17 Use of a pharma- ceutically acceptable salt as defined in any one of embodiments 1.1 to 1.48 for the manufacture of a medicament for the treatment of a patient suffering from a cancer characterized by defects in DNA repair mechanisms, cell cycle defects, or high levels of replication stress.

[0141] 3.18 The use according to embodiment 3.17, wherein said cancer is a p53-negative or p53-mutated tumor.

[0142] 3.19 A method of treating a patient (e.g., a human patient) with a cancer characterized by defective DNA repair mechanisms, cell cycle defects, or high levels of replication stress, comprising administering to the patient a therapeutically effective amount of a pharma- ceutically acceptable salt as defined in any one of embodiments 1.1 to 1.48.

[0143] 3.20 The method of embodiment 3.19, wherein said cancer is a p53-negative or p53-mutated tumor.

[0144] 3.21 A pharma- ceutically acceptable salt, use, or method for use as defined in any one of embodiments 3.3 to 3.10, wherein said cancer is a RAD17 mutant tumor or an ATM-deficient RAD50 mutant tumor.

[0145] 3.21 A pharma- ceutically acceptable salt as defined in any one of embodiments 1.1 to 1.48 for use in treating Fanconi anemia.

[0146] 3.22 The use of a pharma- ceutically acceptable salt as defined in any one of embodiments 1.1 to 1.48 for the manufacture of a medicament for the treatment of Fanconi anemia.

[0147] 3.23 A method of treating Fanconi anemia in a subject (e.g., a human subject) in need of such treatment, comprising administering to the subject a therapeutically effective amount of a pharma- ceutically acceptable salt as defined in any one of embodiments 1.1 to 1.48.

[0148] The Chk-1 inhibitor salts of the present invention may be used alone or in combination with DNA damaging anti-cancer drugs and / or radiation therapy and / or immunotherapy to treat subjects suffering from multidrug resistant cancer. A cancer is considered resistant to a drug if the tumor initially responds to the drug and then the tumor growth rate returns to normal during treatment with the drug. A tumor is considered to be "responsive to a drug" if it shows a decrease in tumor mass or a decrease in tumor growth rate.

[0149] Prior to administration of a pharma- ceutically acceptable salt as defined in any one of embodiments 1.1 to 1.48, the patient may be screened to determine whether the cancer that the patient has or may have is a cancer that is susceptible 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.

[0150] More particularly, patients may be screened to determine whether the cancer they have or may have is a cancer characterized by defects in DNA repair mechanisms, cell cycle defects, or high levels of replication stress, e.g., cell cycle defects due to p53 mutations, or whether it is a p53-negative cancer.

[0151] Cancers characterized by p53 mutations or lack 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 in the articles cited in the introduction to this application. For example, p53 protein can be detected by immunohistochemical methods, such as immunostaining.

[0152] Diagnostic tests are typically performed on a biological sample selected from a tumor biopsy, a blood sample (isolation and enrichment of sloughed tumor cells), a stool biopsy, sputum, chromosome analysis, pleural effusion, peritoneal fluid, or urine.

[0153] Besides p53, mutations in other DNA repair factors such as RAD17, RAD50, and members of the Fanconi anemia complementation group may predict response to Chk1 inhibitors alone or in combination with chemotherapy. Cancers containing mutations in these DNA repair pathways may 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 focus formation in tumor biopsy specimens using an antibody against FANCD2, as described in Duan et al., Frontiers in Oncology vol.4,1-8 (2014).

[0154] Thus, a pharma- ceutically acceptable salt as defined in any one of embodiments 1.1 to 1.48 may be used to treat members of a subpopulation of patients who have been screened (e.g., by testing one or more biological samples taken from the patient) and found to be afflicted with a cancer characterized by a p53 mutation or a p53-negative cancer, or a cancer that includes a RAD17 mutation or a RAD50 mutation, or a mutation in a member of the Fanconi anemia complementation group.

[0155] Thus, in further embodiments (embodiment 3.24 to embodiment 3.30), the present invention provides:

[0156] 3.24 A pharmaceutically acceptable salt as defined in any one of embodiments 1.1 to 1.48 for use in treating cancer in a subject (e.g., a human subject) who has been screened and determined to be afflicted with a cancer that would 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.

[0157] 3.25 A pharmaceutically acceptable salt as defined in any one of embodiments 1.1 to 1.48 for use in treating cancer in a subject (e.g., a human subject) who has been screened and determined to be afflicted with a cancer characterized by a DNA repair defect or a cell cycle defect, e.g. a cell cycle defect due to a p53 mutation, or which is a p53 negative cancer.

[0158] 3.26 A pharmaceutically acceptable salt as defined in any one of embodiments 1.1 to 1.48 for use in treating cancer in a subject (e.g., a human subject) screened and determined to be afflicted with a cancer characterized by a p53 mutation or a p53-negative cancer, or a cancer comprising a RAD17 mutation or a RAD50 mutation, or a mutation in a member of the Fanconi anemia complementation group.

[0159] 3.27 Use of a pharma- ceutically acceptable salt as defined in any one of embodiments 1.1 to 1.48 for the manufacture of a medicament for the use as defined in any one of embodiments 3.24 to 3.26.

[0160] 3.28 A method of treating cancer in a subject (e.g., a human subject) who has been screened and determined to have a cancer that would 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, comprising administering a therapeutically effective amount of a pharmaceutically acceptable salt as defined in any one of embodiments 1.1 to 1.48, and optionally a chemotherapeutic agent (such as a DNA damaging agent).

[0161] 3.29 A method of treating cancer in a subject (e.g., a human subject) who has been screened and determined to be afflicted with a cancer characterized by a DNA repair defect or a cell cycle defect, e.g., a cell cycle defect due to a p53 mutation or which is a p53 negative cancer, comprising administering a therapeutically effective amount of a pharmaceutically acceptable salt as defined in any one of embodiments 1.1 to 1.48.

[0162] 3.30 A method of treating cancer in a subject (e.g., a human subject) screened to be afflicted with a cancer characterized by a p53 mutation or a p53-negative cancer, or a cancer comprising a RAD17 mutation or a RAD50 mutation, or a mutation in a member of the Fanconi anemia complementation group, comprising administering to the subject a therapeutically effective amount of a pharma- ceutical acceptable salt as defined in any one of embodiments 1.1 to 1.48.

[0163] (Combination therapy) The pharma- ceutically acceptable salts as defined in any one of embodiments 1.1 to 1.48 are believed to be useful, alone or in combination with chemotherapeutic agents (particularly DNA damaging agents), radiotherapy, or immunotherapy, in the prophylaxis or treatment of various proliferative disease conditions or pathologies. Examples of such conditions and pathologies are described above.

[0164] The pharma- ceutically acceptable salts as defined in any one of embodiments 1.1 to 1.48, whether administered alone or in combination with DNA damaging agents or other anti-cancer agents and therapies, are generally administered to a subject in need of such administration, e.g., a human or animal patient, preferably a human.

[0165] According to another embodiment of the present invention, embodiment 4.1, there is provided a combination of a pharma- ceutically acceptable salt as defined in any one of embodiments 1.1 to 1.48 with another chemotherapeutic agent, e.g. an anti-cancer agent.

[0166] Chemotherapeutic agents that may be co-administered with the pharma- ceutically acceptable salts defined in any one of embodiments 1.1 to 1.48 include, for example, the following: Topoisomerase I inhibitors ·Antimetabolites Tubulin-targeting drugs DNA binders and topoisomerase II inhibitors Alkylating agents Monoclonal antibodies Antihormonal drugs Signal transduction inhibitors Proteasome inhibitors DNA methyltransferase Cytokines and retinoids Hypoxia-inducible DNA damaging agents (e.g., tirapazamine, TH-302).

[0167] Particular examples of chemotherapeutic agents that may be administered in combination with a pharma- ceutically acceptable salt as defined in any one of embodiments 1.1 to 1.48 include: Nitrogen mustards, such as mechlorethamine, cyclophosphamide, ifosfamide, melphalan, and chlorambucil; Nitrosoureas such as carmustine, lomustine, and semustine; Ethyleneimine / methylmelamine compounds such as triethylenemelamine, triethylenethiophosphoramide, and hexamethylmelamine; Alkyl sulfonates such as busulfan; triazines such as dacarbazine; antimetabolites such as folate, methotrexate, trimetrexate, 5-fluorouracil, fluorodeoxyuridine, gemcitabine, cytosine arabinoside, 5-azacytidine, 2,2'-difluorodeoxycytidine, 6-mercaptopurine, 6-thioguanine, azathioprine, 2'-deoxycoformycin, erythrohydroxynonyl-adenine, fludarabine phosphate, and 2-chlorodeoxyadenosine; Type I topoisomerase inhibitors such as camptothecin, topotecan, and irinotecan; Type II topoisomerase inhibitors, such as epipodophyllotoxins (e.g., etoposide and teniposide); Antimitotic agents such as paclitaxel, taxotere, vinca alkaloids (e.g., vinblastine, vincristine, vinorelbine), and estramustine (e.g., estramustine phosphate); Antibiotics such as actinomycin D, daunomycin (rubidomycin), doxorubicin (adriamycin), mitoxantrone, idarubicin, bleomycin, mithramycin, mitomycin C, and dactinomycin; Enzymes such as L-asparaginase; Cytokines and biological response modifiers, such as interferons (alpha, beta, gamma), interleukin-2, G-CSF, and GM-CSF: Retinoids, such as retinoic acid derivatives (e.g., bexarotene); Radiosensitizers such as metronidazole, misonidazole, desmethylmisonidazole, pimonidazole, etanidazole, nimodazole, nicotinamide, 5-bromodeoxyuridine, 5-iododeoxyuridine, and bromodeoxycytidine; Platinum compounds such as cisplatin, carboplatin, spiroplatin, iproplatin, onnaplatin, tetraplatin, and oxaliplatin; anthracenediones such as mitoxantrone; ureas such as hydroxyurea; Hydrazine derivatives such as N-methylhydrazine and procarbazine; Adrenal cortical suppressants such as mitotane and aminoglutethimide; Corticosteroids and antagonists, such as prednisone, dexamethasone, and aminoglutethimide; Progestins such as hydroxyprogesterone (e.g., hydroxyprogesterone caproate), medroxyprogesterone (e.g., medroxyprogesterone acetate), and megestrol (e.g., megestrol acetate); estrogens, such as diethylstilbestrol and ethinyl estradiol; anti-estrogens such as tamoxifen; Androgens, such as testosterone (e.g., testosterone propionate) and fluoxymesterone; antiandrogens such as flutamide and leuprolide; Nonsteroidal antiandrogens, such as flutamide; and Signal transduction inhibitors such as PARP inhibitors [e.g., those disclosed in Cancer Res.; 66: (16)], Mek inhibitors [e.g., those disclosed in Blood. 2008; 112(6): 2439-2449], farnesyltransferase inhibitors [e.g., those disclosed in Blood. 2005 Feb. 15; 105(4): 1706-16], wee1 inhibitors [e.g., those disclosed in Haematologica 2014, 99(4): 68], rapamycin, and Src inhibitors [e.g., those disclosed in Blood. 2011 Feb. 10; 117(6): 1947-57]. Immunotherapeutic agents such as anti-PD-L1 [e.g., those disclosed in Cancer Discov. 2019(5):646]

[0168] Chemotherapeutic agents that may be used in combination with the pharma- ceutically acceptable salts as defined in any one of embodiments 1.1 to 1.48 include, for example, 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.

[0169] Specific examples of chemotherapeutic agents that may be used in combination with a pharma- ceutically acceptable salt as defined in any one of embodiments 1.1 to 1.48 include antimetabolites (such as capecitabine, cytarabine, fludarabine, gemcitabine, and pemetrexed), topoisomerase I inhibitors (such as SN38, topotecan, irinotecan), platinum compounds (such as carboplatin, oxaloplatin, and cisplatin), topoisomerase II inhibitors (such as daunorubicin, doxorubicin, 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).

[0170] An additional series of chemotherapeutic agents that may be used in combination with the pharma- ceutically acceptable salts defined in any one of embodiments 1.1 to 1.48 include agents that induce stalling of replication forks (see Ashwell et al., Clin. Cancer Res., supra); such compounds include, for example, gemcitabine, 5-fluorouracil, and hydroxyurea.

[0171] (Pothology) A pharma- ceutically acceptable salt as defined in any one of embodiments 1.1 to 1.48 or a therapeutic combination as defined in embodiment 4.1 is administered to a patient in need thereof (e.g., a human or animal patient) in an amount sufficient to achieve the desired therapeutic effect, such as those described above in embodiments 3.1 to 3.30.

[0172] A pharma- ceutically acceptable salt as defined in any one of embodiments 1.1 to 1.48 or a therapeutic combination as defined in embodiment 4.1 is generally administered to a subject in need of such administration, e.g. a human or animal patient, preferably a human.

[0173] The pharmaceutically acceptable salt of the present invention or the therapeutic combination of the present invention as defined in any one of embodiment 1.1 to embodiment 1.48 is typically administered in an amount that is therapeutically or prophylactically useful and generally non-toxic.However, in certain circumstances, the benefits of administering the pharmaceutically acceptable salt of the present invention or the therapeutic combination of the present invention as defined in embodiment 4.1 may outweigh the drawbacks of toxic effects or side effects, and in this case, it may be considered desirable to administer the pharmaceutically acceptable salt of the present invention or the therapeutic combination of the present invention as defined in embodiment 4.1 in an amount that is associated with some toxicity.

[0174] The pharma- ceutically acceptable salts as defined in any one of embodiments 1.1 to 1.48, and the combination with chemotherapeutic agents or radiotherapy as described and defined above (e.g., as in embodiment 4.1), may be administered chronically or for a short period of time to maintain a beneficial therapeutic effect, and may be administered periodically or continuously.

[0175] The pharma- ceutically acceptable salt as defined in any one of embodiments 1.1 to 1.48 or the combination for treatment as defined in embodiment 4.1 is administered alone (in monotherapy) or in combination with one or more chemotherapeutic agents or radiation therapy in an effective amount, i.e., an amount effective to produce the desired therapeutic effect. For example, an "effective amount" can be an amount of a pharma- ceutically acceptable salt that slows tumor growth, improves disease symptoms, and / or extends lifespan when administered alone or together with a DNA damaging agent or other anti-cancer agent to a subject suffering from cancer. More specifically, an effective amount of the pharma- ceutical acceptable salt of the present invention when used in combination with radiation therapy, or a DNA damaging agent or other anti-cancer agent, is an amount that results in a greater response when the pharma- ceutical acceptable salt is co-administered with a DNA damaging anti-cancer agent and / or radiation therapy compared to when the DNA damaging anti-cancer agent and / or radiation therapy is administered alone. When used as a combination therapy, an "effective amount" of a DNA damaging agent and / or an "effective" amount of radiation is administered to the subject, which is an amount that normally results in an anti-cancer effect. The pharma- ceutically acceptable salt as defined in any one of embodiments 1.1 to 1.48 and the DNA damaging anticancer drug may be co-administered to a subject as part of the same pharmaceutical composition or as separate pharmaceutical compositions.

[0176] When administered as separate pharmaceutical compositions, the pharma- ceutically acceptable salt as defined in any one of embodiments 1.1 to 1.48 and the DNA damaging anticancer drug (and / or radiation therapy) may be administered simultaneously or at different times, so long as the enhancing effect of the pharma- ceutically acceptable salt as defined in any one of embodiments 1.1 to 1.48 is maintained.

[0177] In one embodiment, the pharma- ceutically acceptable salt as defined in any one of embodiments 1.1 to 1.48 is administered prior to (e.g., up to 8 hours, up to 12 hours, or up to 1 day prior to) administration of the DNA damaging anticancer drug.

[0178] In another embodiment, the pharma- ceutically acceptable salt as defined in any one of embodiments 1.1 to 1.48 is administered (e.g., up to 8 hours, up to 12 hours, up to 24 hours, up to 30 hours, or up to 48 hours) after administration of the DNA damaging anticancer drug. In another embodiment, a first administration of the pharma- ceutically acceptable salt as defined in any one of embodiments 1.1 to 1.48 is administered one day after administration of the DNA damaging anticancer drug, and a second administration of the compound is administered two days after administration of the DNA damaging anticancer drug.

[0179] In a further embodiment, a first administration of a pharma- ceutically acceptable salt as defined in any one of embodiments 1.1 to 1.48 is administered one day after administration of the DNA damaging anticancer agent, a second administration is administered two days after administration of the DNA damaging anticancer agent, and a third administration is administered three days after administration of the DNA damaging anticancer agent.

[0180] A particular dosing regimen comprising administration of a pharma- ceutically acceptable salt as defined in any one of embodiments 1.1 to 1.48 and a DNA damaging anticancer drug may be as described in WO2010 / 118390 (Array BioPharma), the contents of which are incorporated herein by reference.

[0181] The amount of the pharma- ceutically acceptable salt of the present invention and the amount of DNA damaging anticancer drug (in case of combination therapy) and radiation dose administered to a subject depend on the nature and potency of the DNA damaging anticancer drug, the type and severity of the disease or condition, and the characteristics of the subject, such as general health, age, sex, weight, and drug resistance. Those skilled in the art will be able to determine the appropriate dose depending on these and other factors. The effective doses of commonly used anticancer drugs and radiation therapy are well known to those skilled in the art.

[0182] A typical daily dosage of the pharma- ceutically acceptable salts as defined in any one of embodiments 1.1 to 1.48, whether administered alone in a monotherapy or in combination with a DNA damaging anticancer drug, may range from 100 picograms to 100 milligrams per kilogram of body weight, more typically from 5 nanograms to 25 milligrams per kilogram of body weight, more usually from 10 nanograms to 15 milligrams per kilogram of body weight (e.g., from 10 nanograms to 10 milligrams, more typically from 1 microgram per kilogram to 20 milligrams per kilogram, e.g., from 1 microgram to 10 milligrams per kilogram). However, higher or lower dosages may be administered as needed. The compounds may be administered daily or repeatedly, for example, every 2, 3, 4, 5, 6, 7, 10, 14, 21, or 28 days.

[0183] Ultimately, however, the dosage of the pharma- ceutically acceptable salt and type of composition used will be commensurate with the disease or physiological condition being treated and will be left to the discretion of the physician.

[0184] (Pharmaceutical preparations) The pharmaceutically acceptable salt as defined in any one of embodiments 1.1 to 1.48 and the therapeutic combination as defined in embodiment 4.1 are typically administered to patients in the form of a pharmaceutical composition.Accordingly, in another embodiment (embodiment 5.1) of the present invention, the present invention provides a pharmaceutical composition comprising a pharmaceutically acceptable salt as defined in any one of embodiments 1.1 to 1.48, a pharmaceutically acceptable excipient, and optionally a further chemotherapeutic agent.

[0185] In further embodiments, the following is provided:

[0186] 5.2 The pharmaceutical composition according to embodiment 5.1, comprising from about 1% (w / w) to about 95% (w / w) of a pharma- ceutically acceptable salt as defined in any one of embodiments 1.1 to 1.48, and from 99% (w / w) to 5% (w / w) of a pharma- ceutically acceptable excipient or combination of excipients, and optionally one or more further therapeutically active ingredients.

[0187] 5.3 A pharmaceutical composition according to embodiment 5.2, comprising from about 5% (w / w) to about 90% (w / w) of a pharma- ceutically acceptable salt composition as defined in any one of embodiments 1.1 to 1.48, and from 95% (w / w) to 10% of a pharmaceutical excipient or combination of excipients, and optionally one or more further therapeutically active ingredients.

[0188] 5.4 The pharmaceutical composition according to embodiment 5.3, comprising from about 10% (w / w) to about 90% (w / w) of a pharma- ceutically acceptable salt as defined in any one of embodiments 1.1 to 1.48, and from 90% (w / w) to 10% of a pharmaceutical excipient or combination of excipients.

[0189] 5.5 The pharmaceutical composition according to embodiment 5.4, comprising from about 20% (w / w) to about 90% (w / w) of a pharma- ceutically acceptable salt as defined in any one of embodiments 1.1 to 1.48, and from 80% (w / w) to 10% of a pharmaceutical excipient or combination of excipients.

[0190] 5.6 The pharmaceutical composition according to embodiment 5.5, comprising from about 25% (w / w) to about 80% (w / w) of a pharma- ceutically acceptable salt as defined in any one of embodiments 1.1 to 1.48, and from 75% (w / w) to 20% of a pharmaceutical excipient or combination of excipients.

[0191] The pharmaceutical compositions of the present invention can be in any suitable form for oral, parenteral, topical, intranasal, intrabronchial, ocular, otic, rectal, vaginal or transdermal administration. If the composition is intended for parenteral administration, it can be formulated for intravenous, intramuscular, intraperitoneal, subcutaneous administration, or for direct delivery to a target organ or tissue by injection, infusion or other delivery means.

[0192] Pharmaceutical dosage forms suitable for oral administration include tablets, capsules, caplets, pills, lozenges, syrups, solutions, sprays, powders, granules, elixirs and suspensions, sublingual tablets, sprays, wafers, or patches and buccal patches.

[0193] Thus, in further embodiments, the present invention provides:

[0194] 5.7 A pharmaceutical composition according to any one of embodiments 5.1 to 5.6, suitable for oral administration.

[0195] 5.8 The pharmaceutical composition according to embodiment 5.7, selected from tablets, capsules, caplets, pills, lozenges, syrups, solutions, sprays, powders, granules, elixirs and suspensions, sublingual tablets, sprays, wafers, or patches and buccal patches.

[0196] 5.9 The pharmaceutical composition according to embodiment 5.8, selected from a tablet and a capsule.

[0197] 5.10 A pharmaceutical composition according to any one of embodiments 5.1 to 5.6, which is suitable for parenteral administration.

[0198] 5.11 The pharmaceutical composition of embodiment 5.10, formulated for intravenous, intramuscular, intraperitoneal, subcutaneous administration, or for direct delivery to a target organ or tissue by injection, infusion, or other delivery means.

[0199] 5.12 The pharmaceutical composition according to embodiment 5.11, which is a solution or suspension for injection or infusion.

[0200] Pharmaceutical compositions containing the pharma- ceutically acceptable salts as defined in any one of embodiments 1.1 to 1.48 (e.g., as defined in any one of embodiments 5.1 to 5.12) can be formulated according to known techniques. See, for example, Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, Pennsylvania, USA.

[0201] Thus, a tablet composition (such as embodiment 5.9) may contain a unit dose of a pharma- ceutically acceptable salt as defined in any one of embodiments 1.1 to 1.48, together with an inert diluent or carrier, such as a sugar or sugar alcohol, for example lactose, sucrose, sorbitol, or mannitol, and / or a non-sugar derived diluent, such as sodium carbonate, calcium phosphate, talc, calcium carbonate, or a cellulose or derivative thereof, such as methylcellulose, ethylcellulose, hydroxypropylmethylcellulose, and a starch, such as cornstarch. Tablets may contain standard ingredients such as binders and granulating agents, such as polyvinylpyrrolidone, disintegrants (e.g., swellable cross-linked polymers, such as cross-linked carboxymethylcellulose), lubricants (e.g., stearates), preservatives (e.g., parabens), antioxidants (e.g., BHT), buffers (e.g., phosphate buffers or citrate buffers), and effervescent agents, such as citrate / bicarbonate mixtures. Such excipients are well known and need not be described in detail herein.

[0202] Capsule formulations (such as embodiment 5.9) may be of the hard or soft gelatin variety and may contain the active ingredient in solid, semi-solid, or liquid form. Gelatin capsules may be formed from animal gelatin or its synthetic or vegetable-derived equivalents.

[0203] The solid dosage forms (e.g., tablets, capsules, etc.) may be coated or uncoated, but typically have a coating, such as a protective film coating (e.g., wax or varnish) or a sustained release coating. The coating (e.g., Eudragit™ type polymer) can be designed to release the pharmaceutically acceptable salt at a desired location in the gastrointestinal tract. Thus, the coating can be selected to degrade under certain pH conditions in the gastrointestinal tract, thereby selectively releasing the pharmaceutically acceptable salt in the stomach or ileum or duodenum.

[0204] Instead of, or in addition to, a coating, the drug can be provided in a solid matrix containing a release controlling agent, e.g., a release retarding agent, which can be adapted to selectively release a pharma- ceutically acceptable salt under conditions of varying acidity or alkalinity within the gastrointestinal tract. Alternatively, such a matrix material or release retarding coating can be in the form of an erodible polymer (e.g., a maleic anhydride polymer) that erodes substantially continuously as the dosage form passes through the gastrointestinal tract.

[0205] Compositions for topical use include ointments, creams, sprays, patches, gels, drops, and inserts (e.g., intraocular inserts). Such compositions can be formulated according to conventional methods.

[0206] Compositions for parenteral administration (such as those of embodiments 5.10 to 5.12) are typically provided as sterile aqueous or oily solutions or sterile fine suspensions, but may also be provided in finely divided sterile powder form for extemporaneous reconstitution with sterile water for injection.

[0207] Examples of formulations for rectal or vaginal administration include pessaries and suppositories which may be formed, for example, from a moldable or waxy material containing the active compound.

[0208] The composition for inhalation administration can be in the form of inhalable powder composition or liquid spray or powder spray, and can be administered in standard form using powder inhalation device or aerosol dispensing device.Such devices are well known.For inhalation administration, powdered preparations typically comprise the above-mentioned pharmaceutically acceptable salt together with powdered inert solid diluent such as lactose.

[0209] The pharmaceutical compositions are generally provided in unit dosage form and, as such, typically contain sufficient pharma- ceutically acceptable salt to provide a desired level of biological activity. For example, a pharmaceutical composition intended for oral administration, such as a pharmaceutical composition according to any one of embodiments 5.1 to 5.9, may contain from 2 milligrams to 200 milligrams of the pharma- ceutically acceptable salt, more usually from 10 milligrams to 100 milligrams, e.g., 12.5 milligrams, 25 milligrams, or 50 milligrams.

[0210] The pharmaceutical composition may optionally comprise a further chemotherapeutic agent as defined in embodiment 4.1.

[0211] Thus, in a further embodiment (embodiment 5.13), the present invention provides a pharmaceutical composition as defined in any one of embodiments 5.2 to 5.12, additionally comprising a further chemotherapeutic agent as defined in embodiment 4.1. [Brief description of the drawings]

[0212] [Figure 1] FIG. 1 is an XRPD spectrum of the free base of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile ("compound of formula (1)"). [Diagram 2] FIG. 2 shows the DSC and TGA traces of the free base of the compound of formula (1). [Diagram 3]FIG. 3 shows the GVS profile of the free base of the compound of formula (1). [Figure 4] FIG. 4 shows the XRPD spectra of several crystalline forms of the free base (top trace) and hydrochloride salt of the compound of formula (1), with the crystalline forms of the salt being Pattern A, Pattern B, Pattern C, Pattern D, and Pattern E, respectively, from the second trace from the top to the bottom trace. [Diagram 5] FIG. 5 shows the XRPD spectra of several crystalline forms of the free base (top trace) and hydrobromide salt of the compound of formula (1), with the crystalline forms of the salt being Pattern A, Pattern B, Pattern C, and Pattern D, respectively, from the second trace from the top to the bottom trace. [Figure 6] FIG. 6 shows the XRPD spectra of several crystalline forms of the free base (top trace) and mesylate salt of the compound of formula (1), with the crystalline forms of the salt being Pattern A, Pattern B, and Pattern C, respectively, from the second trace from the top to the bottom trace. [Figure 7] FIG. 7 shows the XRPD spectra of several forms of the free base (top trace) and the L-tartrate salt of the compound of formula (1), from the second trace to the bottom trace, respectively, of the salt in the amorphous form (second trace), Pattern A (third trace from the top), and Pattern B (bottom trace). [Figure 8] FIG. 8 shows the XRPD spectra of several crystalline forms of the free base (top trace) and esylate salt of the compound of formula (1), with each crystalline form of the salt being Pattern A (second trace from the top) and Pattern B (third and fourth traces from the top), respectively. [Figure 9] FIG. 9 shows the XRPD spectra of the free base (top trace) and the L-aspartate salt crystalline form (bottom trace) of the compound of formula (1). [Figure 10] FIG. 10 shows the XRPD spectra of several crystalline forms of the besylate salt of the compound of formula (1), from top to bottom, Pattern A, Pattern B, and Pattern C for each crystalline form of the salt. [Figure 11] FIG. 11 shows the XRPD spectra of several crystalline forms of the tosylate salt of the compound of formula (1), which are, from top to bottom, Pattern A, Pattern B, Pattern C, and Pattern D, respectively. [Figure 12] FIG. 12 shows XRPD spectra of several crystalline forms of the free base and sulfate salt of the compound of formula (1), from top to bottom: the free base (top trace), salt pattern A (second and third traces from the top), and salt pattern B (bottom trace); [Figure 13] FIG. 13 shows XRPD spectra of several crystalline forms of the free base and phosphate salt of the compound of formula (1), from top to bottom: the free base (top trace), salt pattern A (second and third traces from the top), and salt pattern B (bottom trace). [Figure 14] FIG. 14 shows the XRPD spectra of several amorphous and crystalline forms of the free base and citrate salt of the compound of formula (1); from top to bottom, the traces are for the free base (top trace), the amorphous salt (second trace from the top), and the Pattern A and Pattern B salts. [Figure 15] FIG. 15 shows the XRPD spectra of several crystalline forms of the free base and acetate salt of the compound of formula (1); from top to bottom, the traces are the free base (top trace), salt pattern A, and salt pattern B. [Figure 16] FIG. 16 shows the XRPD spectra of the free base (top trace) and the Pattern A crystal form of the L-glutamate salt (bottom trace) of the compound of formula (1). [Figure 17] FIG. 17 shows the XRPD spectra of several crystalline forms of the maleate salt of the compound of formula (1), from top to bottom being traces of Pattern A, Pattern B, and Pattern C. [Figure 18] FIG. 18 shows the XRPD spectra of the free base (top trace) and the Pattern A crystal form of the gentisate salt of the compound of formula (1) (middle and bottom traces). [Figure 19]FIG. 19 shows the XRPD spectra of several crystalline forms of the free base (top trace) and glucuronate salt of the compound of formula (1) (Pattern A--middle trace, Pattern B--bottom trace). [Figure 20] FIG. 20 shows the XRPD spectra of several crystalline forms of the free base (top trace) and malonate salt of the compound of formula (1) (Pattern A--middle trace, Pattern B--bottom trace). [Figure 21] FIG. 21 shows the XRPD spectra of the crystalline form of the naphthalene-2-sulfonate salt of the compound of formula (1) isolated from THF (top trace) and THF:H20 (bottom trace). [Figure 22] FIG. 22 shows the XRPD spectra of several crystalline forms of the free base (top trace) and oxalate salt of the compound of formula (1) (Pattern A-middle trace), and Pattern B (bottom trace). [Figure 23] FIG. 23 shows the XRPD spectra of crystalline forms A, B, C, and D (in descending order from the second top) of the free base (top trace) and sulfate salt of the compound of formula (1). [Figure 24] FIG. 24 shows the XRPD spectra of the free base (top trace) and sulfate salt crystalline forms D and E (middle and bottom traces) of the compound of formula (1). [Diagram 25] FIG. 25 shows the XRPD spectrum of the maleic acid Pattern B salt. [Figure 26] FIG. 26 shows the DSC and TGA traces of the maleic acid Pattern B salt. [Figure 27] FIG. 27 shows the XRPD spectrum of maleic acid Pattern A salt. [Figure 28] FIG. 28 shows the DSC and TGA traces of the maleic acid Pattern A salt. [Figure 29] FIG. 29 shows the XRPD spectrum of maleic acid Pattern C salt. [Diagram 30] FIG. 30 shows the DSC and TGA traces of the maleic acid Pattern C salt. [Diagram 31] FIG. 31 shows the XRPD spectrum of the malonic acid Pattern B salt. [Diagram 32] FIG. 32 shows the DSC and TGA traces of the malonic acid Pattern B salt. [Diagram 33] FIG. 33 shows the XRPD spectrum of the tosylate Pattern A salt. [Diagram 34] FIG. 34 shows the DSC and TGA traces of the tosylate Pattern A salt. [Diagram 35] FIG. 35 shows the XRPD spectrum of the besylate pattern C salt. [Diagram 36] FIG. 36 shows the DSC trace and the TGA trace of the besylate Pattern C salt of the compound of formula (1). [Figure 37] FIG. 37 shows the XRPD spectra of the free base (top trace) and bis-mesylate salt crystalline forms Pattern A (middle trace) and Pattern B (bottom trace) of the compound of formula (1). [Figure 38] FIG. 38 shows the XRPD spectra of the free base (top trace) and the bis-maleate salt crystalline forms Pattern A (middle trace) and Pattern B (bottom trace) of the compound of formula (1). [Figure 39] FIG. 39 shows the XRPD spectra of the free base (top trace) and the bis-besylate salt crystalline forms Pattern A (middle trace) and Pattern B (bottom trace) of the compound of formula (1). [Diagram 40] FIG. 40 shows the XRPD spectra of various crystalline forms of the free base and maleate salts, which, from the top trace to the bottom trace, are, in descending order, the free base, Pattern A mono-maleate salt, Pattern A bis-maleate salt, Pattern B bis-maleate salt, and Pattern A hemi-maleate salt of the compound of formula (1). [Diagram 41] FIG. 41 shows the XRPD spectra of the free base (top trace) and the hemi-ethane-1,2-disulfonic acid salt crystalline form Pattern A (bottom trace). [Diagram 42]FIG. 42 shows the XRPD spectra of the free base (top trace) and the hemi-naphthalene-1,5-disulfonic acid salt crystalline form Pattern A (bottom trace). [Diagram 43] FIG. 43 shows the XRPD spectra of various crystalline forms of the free base and the hemi-fumarate salt, in descending order from the top trace to the bottom trace, of the free base, Pattern A hemi-fumarate salt, Pattern B hemi-fumarate salt, and Pattern C hemi-fumarate salt of the compound of formula (1). [Diagram 44] FIG. 44 shows a Gravimetric Vapor Sorption (GVS) plot of the Pattern A crystal form of the maleate salt of the compound of formula (1). [Diagram 45] FIG. 45 shows a GVS plot of the Pattern B crystal form of the maleate salt of the compound of formula (1). [Figure 46] FIG. 46 shows a GVS plot of the Pattern A crystal form of the tosylate salt of the compound of formula (1). [Figure 47] FIG. 47 shows a GVS plot of the Pattern A crystal form of the besylate salt of the compound of formula (1). [Figure 48] FIG. 48 shows a GVS plot of the Pattern B crystal form of the besylate salt of the compound of formula (1). [Figure 49] FIG. 49 shows a GVS plot of the Pattern C crystal form of the besylate salt of the compound of formula (1). [Figure 50] FIG. 50 shows a GVS plot of the Pattern A crystal form of the naphthalene-2-sulfonate salt of the compound of formula (1). [Figure 51] FIG. 51 shows a GVS plot of the Pattern B crystal form of the malonate salt of the compound of formula (1). [Figure 52] FIG. 52 shows the XRPD patterns of various crystalline forms of the maleate salt, which, from top to bottom, are Pattern A, Pattern B, a mixture of A / B, Pattern C, Pattern D, and Pattern E. [Diagram 53] FIG. 53 shows a DVS plot of the Pattern B crystal form of the Maleate Salt. [Figure 54] FIG. 54 shows the DSC and TGA traces of the maleic acid Pattern D salt. [Figure 55] FIG. 55 shows the DSC and TGA traces of the maleic acid Pattern E salt. EXAMPLES

[0213] (Analysis method) (Proton NMR) The formation of salts (by observing proton shifts versus the free base) and their identification as 1:1 (molar ratio of free base:acid) stoichiometric salts were confirmed by the analysis of the data collected using a JEOL ECX 400 MHz spectrometer equipped with an autosampler. 1 The H NMR spectrum was confirmed. Samples were dissolved in appropriate deuterated solvents for analysis. Data were acquired using Delta NMR Processing and Control Software version 4.3.

[0214] (X-ray powder diffraction (XRPD)) Powder X-ray diffraction patterns were collected on a PANalytical diffractometer using CuKα radiation (45 kV, 40 mA), a θ-θ goniometer, a focusing mirror, a divergence slit (1 / 2''), Soller slits (4 mm) on both the incident and divergent beam sides, and a PIXcel detector. The software used for data collection was X'Pert Data Collector, version 2.2f, and data were displayed using X'Pert Data Viewer, version 1.2d. XRPD patterns were acquired under ambient conditions using a PANalytical X'Pert PRO through a sample stage made of a transparent foil (12.7 μm thick polyimide "Kapton" film). The data collection range was 2.994-35° 2θ, with a continuous scan speed of 0.202004° per second.

[0215] (Differential Scanning Calorimetry (DSC)) DSC data were collected on a PerkinElmer Pyris 6000 DSC equipped with a 45-position sample holder. Certified indium was used to validate the instrument and perform energy and temperature calibrations. A predetermined amount of sample, between 0.5 and 3.0 mg, was placed in an aluminum pan with a pinhole and heated at 20 °C per minute from 30 °C to 350 °C or varied depending on the experiment. A purge of 20 ml per minute of dry nitrogen was maintained over the sample. Instrument control, data acquisition, and analysis were performed using Pyris Software v11.1.1 revision H.

[0216] (Thermogravimetric analysis (TGA)) TGA data were collected on a Perkin Elmer Pyris 1 TGA equipped with a 20-position autosampler. The instrument was verified and temperature calibrated using certified weights and certified Alumel and Perkalloy. Predetermined amounts of sample, 1-5 mg, were loaded into pre-tared aluminum crucibles and heated from ambient to 400 °C at 20 °C per minute. A nitrogen purge of 20 ml per minute was maintained over the sample. Instrument control, data acquisition, and analysis were performed using Pyris Software v11.1.1 revision H.

[0217] (Gravimetric Vapor Sorption (GVS)) GVS studies were carried out on the salts of the invention using the following protocol.

[0218] Sorption isotherms were obtained using a Hiden Isochema moisture sorption analyzer (model IGAsorp) controlled by IGAsorp Systems Software V6.50.48. The instrument controls maintained the samples at a constant temperature (25°C). Humidity was controlled by mixing dry and wet nitrogen flows, with a total flow rate of 250 ml per minute. The instrument was verified for relative humidity (RH) content by measuring three calibrated Rotronic salt solutions (10-50-88%). Sample weight change was monitored as a function of humidity by a microbalance (accuracy + / - 0.005 mg). A pre-determined amount of sample was placed in a tared mesh stainless steel basket under ambient conditions. A complete experimental cycle typically consisted of three scans (sorption, desorption, and sorption) at 10% RH intervals (60 min for each humidity level) over the range of 0-90% at constant temperature (25°C). This type of experiment should demonstrate the ability of the sample under study to absorb (or not absorb) moisture over a series of well-determined humidity ranges.

[0219] (HPLC method 1) HPLC analysis was performed on an Agilent 1110 series HPLC system. The column used was an Aquity BEH Phenyl; 30 x 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 by UV at 210 nm. The injection volume was 5 μL, and the following gradient was used:

[0220] [Table 1]

[0221] (HPLC method 2) HPLC analysis was performed on an Agilent 1110 / 1200 series HPLC system. The column used was a Triart C18; 150 x 4.6 mm, particle size 3.0 μm (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 by UV at 302 nm. The injection volume was 5 μL, the column temperature was 40° C., and the following gradient was used:

[0222] [Table 2]

[0223] Example 1 (Preparation and Characterization of 5-[[5-[4-(4-Fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile Free Base) The title compound was prepared according to the method of Example 64, Method L of WO2015 / 20390, the contents of which are incorporated herein by reference, except that the compound was isolated as the free base rather than as the hydrochloride salt. The free base was characterized by X-ray powder diffraction (XRPD), differential scanning calorimetry (DSC), and thermogravimetric analysis (TGA). The XRPD spectrum and the DSC and TGA traces are shown in Figures 1 and 2.

[0224] The free base was shown by XRPD to be crystalline. The DSC thermograph shows a major melting endotherm with an onset of 205.6°C and a peak at 214°C. The TGA thermograph shows a weight loss of 2.8% up to 150°C. The solid 1 The H NMR spectrum is consistent with the molecular structure. Since there is no significant solvent present in the NMR spectrum, the weight loss shown in the TGA thermograph is related to the loss of water upon heating of the material.

[0225] The GVS profile of the free base is shown in Figure 3. During the first desorption cycle, the solid loses 2 wt. % from 50% to 0% relative humidity (RH). During the subsequent sorption cycle, the solid gains 8% water up to 90% RH. The water uptake is reversible and hysteresis is observed. The theoretical water content for the formal monohydrate of the free base is 4.2%, so water is absorbed up to the dihydrate level at the humidity extremes.

[0226] Example 2 (Preparation of Salts) Small Scale Law The acid addition salts of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile were prepared from the free base by small scale methods 1-7 below.

[0227] Method 1: THF-mediated Sixteen crystallization tubes were charged with the free base of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile (50 mg). THF (2 mL, 40 volumes) was added and each resulting mixture was heated to 60°C. The respective acid (1 M, 1 eq) was charged as one aliquot. Each solution was held at temperature and equilibrated for 1 hour. The solutions were then cooled to room temperature and equilibrated for 18 hours before being isolated by filtration and dried under vacuum for 18 hours. In some cases where crystallization did not occur, the samples were further manipulated by solvent removal with nitrogen and trituration of the solid with MeOH. Solvent reduction and trituration was required for the esylate, besylate, acetate, and malonate salts.

[0228] Method 2: THF:MeCN-mediated Method 2 was identical to Method 1, except that a mixture of THF:MeCN (1:1) (1 mL, 20 vol) was used as the solvent and the mixture was heated to 50° C. The benzenesulfonate salt required solvent reduction and trituration.

[0229] Method 3: THF: Waterborne Method 3 was identical to Method 2, except that THF:water (95:5) (1 mL, 20 vol) was used as the solvent and the mixture was heated to 50° C. The benzenesulfonate, acetate, L-glutamate, and L-aspartate salts required solvent reduction and trituration.

[0230] Method 4: THF-mediated, using excess acid Method 4 was identical to Method 1 except that the acid (1 M, 1.84 equiv.) was charged as a single aliquot. Using this method, hydrochloride Pattern A, hemi-fumarate Pattern A, hydrobromide Pattern C, bis-mesylate Pattern A, bis-maleate Pattern A, bis-besylate Pattern A, tosylate Pattern C, and acetate Pattern B were isolated.

[0231] Method 5: THF:water borne, using excess acid Method 5 was identical to Method 2, except that the acid (1 M, 1.84 equiv.) was charged as a single aliquot. Using this method, L-tartrate Pattern B, tosylate Pattern A, phosphate Pattern B, citrate Pattern B, acetate Pattern B, L-glucuronate Pattern A, hydrochloride Pattern D, hydrobromide Pattern D, bis-mesylate Pattern B, bis-maleate Pattern B, besylate Pattern B, sulfate Pattern C were isolated.

[0232] Method 6: THF-mediated, using excess acid Method 6 was identical to Method 1 except that the free base (30 mg) was charged and the acid (1 M, 2 equiv.) was charged in one aliquot. This method was used to generate the hydrochloride Pattern B salt.

[0233] Method 7: THF-mediated, using 0.5 equivalents of acid Method 7 was identical to Method 1 except that 0.5 equivalents of acid was added in all cases. Using Method 7, hemi-maleate Pattern A, hemi-sulfate Pattern A, hemi-ethane-1,2-disulfonate Pattern A, and hemi-naphthylene-1,5-disulfonate Pattern A were isolated.

[0234] (Medium-scale preparation of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile salt) Medium scale method 1 The salts were prepared on a larger scale using similar conditions as used in small scale method 1, except that 300 mg of free base was used. The following salts were prepared by this method: Tosylate pattern C, and thermal cycling at >200°C gave tosylate pattern D. Maleate pattern A was conditioned at 40℃ / 75%RH to obtain maleate pattern B. Besylate Pattern B Naphthalene-2-sulfonate Pattern A

[0235] This method was modified to use 100 mg of the free base to form the following: Oxalate Pattern A

[0236] Medium scale method 2 The preparation of malonic acid pattern B was scaled up according to the following method. 5-[[5-[4-(4-fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile free base (300 mg) was weighed into a 25 mL round bottom flask. THF:water (95:5, 20 vol) was added and the mixture was equilibrated at 60° C. for 15 min. Malonic acid (1 eq) was charged and the mixture was equilibrated at 60° C. for 15 min. The mixture was cooled to room temperature and equilibrated for 30 min. The mixture was then quickly evaporated at 50° C. and 210 rpm using a rotary evaporator. This produced a beige powder. This was then aged in MeOH (20 vol) at room temperature for 18 h. The resulting suspension was isolated by vacuum filtration and the solid was dried in vacuum at 45° C. over the weekend.

[0237] Medium scale method 3 The conditions used in small scale procedure 5 were used except that 300 mg of free base and 2 equivalents of acid were used. This procedure was used to prepare the bis-maleic acid Pattern A salt.

[0238] Medium scale method 4 The conditions used in small scale procedure 4 were used except that 300 mg of free base and 2 equivalents of acid were used. This procedure was used to prepare the bis-besylate salt pattern B.

[0239] Aging method Besylate pattern C was formed after water aging (24 h) of besylate pattern B.

[0240] After water aging of maleate B (24 h), maleate pattern C was formed.

[0241] The methods used to prepare the salts and the physical appearance of the salts so prepared are summarized in the table below.

[0242] [Table 3-1]

[0243] [Table 3-2]

[0244] Data characterizing the salts prepared according to the above procedures are given in the table below.

[0245] [Table 4-1]

[0246] [Table 4-2]

[0247] [Table 4-3]

[0248] [Table 4-4]

[0249] [Table 4-5]

[0250] [Table 4-6]

[0251] [Table 4-7]

[0252] [Table 4-8]

[0253] The characteristics of the bis- and hemi-salts prepared using the above methods are also described below.

[0254] [Table 5-1]

[0255] [Table 5-2]

[0256] [Table 6-1]

[0257] [Table 6-2]

[0258] Example 3A (Measurement of solubility of salt in water) 5-[[5-[4-(4-Fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile free base and selected salts (30 mg) were weighed into crystallizer tubes, charged with water for injection (WFI) (1 mL), and each sample was left to equilibrate (25° C.) for 24 hours. Solids were isolated by vacuum filtration and the filtrate was used to assess solubility by HPLC (HPLC Method 1).

[0259] [Table 7]

[0260] (Conclusion) Although a number of crystalline salts were identified, most of the salts showed a tendency to hydrate / solvate and had complex thermal profiles.The known hydrochloride salt (disclosed in Example 64, Method L of International Patent Application WO2015 / 20390) is not considered to be a good candidate for the preparation of solid formulations, since it shows polymorphism and a poor thermal profile indicating hydration / solvation.

[0261] The tosylate, maleate, besylate, malonate, and oxalate salts all have improved solubility over the free base, but the oxalate salt was not considered for further development due to its poor crystallinity, and the bis-salt was also not considered a candidate for further development due to its disproportionation in water.

[0262] Example 3B (Measurement of solubility of salts in biologically relevant media) experiment: 5-[[5-[4-(4-Fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile free base and selected salts (30 mg) were weighed into crystallization tubes. Biorelevant media (DI HO, FeSSIF, FaSSIF, and FaSSGF) (2 mL) were charged. Samples were left to equilibrate (25 °C) for 24 hours. Solubility was measured using HPLC (see Method 1 above).

[0263] [Table 8]

[0264] Biorelevant solubility evaluation of the free base and selected salts showed low solubilities, generally below 1 mg / mL. A general trend was observed of increasing salt solubility from FaSSIF to FeSSIF to FaSSGF. Maleate and malonate salts showed improved solubility in FaSSGF compared to the free base. Most salts showed similar solubility in FeSSIF and FaSSIF, although differences were observed in gastric fluid for the maleate salt.

[0265] Across the biorelevant range, the salt and free base perform similarly under these test conditions, however the maleate salt shows promise when considering gastrointestinal transit as well as overall solid form performance.

[0266] The hydrochloride salt, although soluble, is polymorphic and has a complex thermal profile (indicating hydration and solvation).

[0267] (2 week stability) (protocol) Experimental: Maleate salt Pattern B (30 mg) was placed into separate 15 mL Type I glass vials. These vials were loosely fitted with HDPE plastic caps to allow for moisture ingress. The vials were then placed in ICH-rated stability cabinets at 25°C / 60% RH and 40°C / 75% RH and refrigerated at 2-8°C. After 2 weeks of storage, the samples were removed from the stability cabinet and refrigerated storage and assessed for chemical purity by HPLC (Method 2). Relevant data were collected at a wavelength of 302 nm. Samples were prepared in MeCN:water (1:1).

[0268] Maleate Pattern B is stable for 2 weeks under the following conditions: 25℃ / 60%RH, 40℃ / 75%RH, and 2~8℃.

[0269] [Table 9]

[0270] The four best performing salts were the maleate, tosylate, besylate, and malonate. Of these, the maleate exhibited the best properties. Selected crystalline forms of these salts are described in more detail below.

[0271] (X-ray powder diffraction study) Maleate Pattern B The XRPD spectrum of the maleate salt Pattern B is shown in Figure 25, and the thermal data is shown in Figure 26. The XRPD peaks for Pattern B are shown in the table below.

[0272] [Table 10]

[0273] Maleate Pattern A The XRPD spectrum of the maleate salt Pattern A is shown in Figure 27, and the thermal data is shown in Figure 28. The XRPD peaks for Pattern A are shown in the table below.

[0274] [Table 11]

[0275] Maleate Pattern C The XRPD spectrum of the maleate salt Pattern C is shown in Figure 29, and the thermal data is shown in Figure 30. The XRPD peaks for Pattern C are shown in the table below.

[0276] [Table 12]

[0277] Malonate Pattern B The XRPD spectrum of malonate Pattern B is shown in Figure 31, and the DSC and TGA traces are shown in Figure 32. The XRPD peaks are shown in the table below.

[0278] [Table 13]

[0279] Tosylate Pattern A The XRPD spectrum of tosylate Pattern A is shown in Figure 33, and the TGA and DSC traces are shown in Figure 34. The XRPD peaks are shown in the table below.

[0280] [Table 14]

[0281] Besylate Pattern C The XRPD spectrum of besylate Pattern C is shown in Figure 35, and the TGA and DSC traces are shown in Figure 36. The XRPD peaks are shown in the table below.

[0282] [Table 15]

[0283] (Gravimetric Vapor Sorption Studies) GVS data obtained using the above protocol for a particular crystalline salt form is set forth below.

[0284] Maleate Pattern A - See Figure 44 During the first sorption cycle, the solid gained 1.5 wt% from 50% to 90% RH. During the subsequent desorption cycle, the solid lost 4% water to 0% RH. This increased to 4 wt% at 90% RH in the next sorption cycle. The GVS profiles confirmed that this water uptake was reversible with decreasing relative humidity and showed only minimal hysteresis.

[0285] At 0% and 90% RH, new patterns were isolated and named B. Pattern B is closely related to pattern A.

[0286] Maleate Pattern B - See Figure 45 During the first desorption cycle, the solid loses 2.5 wt.% from 50% to 0% RH. During the subsequent sorption cycles, the solid gains 4% water up to 90% RH, but there is a sharp increase between 0% and 40% RH. At 0% RH, it transforms to pattern A, and at 90% RH, it remains unchanged. The data suggest that the interconversion between the crystalline versions is related to hydration.

[0287] Tosylate Pattern A - Figure 46 During the first desorption cycle, the solid loses 3.5 wt.% from 50% to 0% RH, then decreases steadily by 0.5 wt.% from 50% to 10% RH, and then shows a rapid decrease of about 3 wt.% from 10% to 0% RH. During the subsequent sorption cycles, the solid rapidly gains about 3% water up to 10% RH, but then gains steadily by about 1% from 10% to 90% RH. The 3% water content corresponds to the monohydrate of the tosylate salt. No morphology change is observed between 0% and 90% RH, suggesting a reversible and stable channel hydrate over the entire ambient range.

[0288] Besylate Pattern A - See Figure 47 During the first desorption cycle, the solid loses 2.5% by weight from 50% to 0% RH. During the subsequent sorption cycles, the solid gains 10% water up to 90% RH, but shows a sudden increase of 6% from 40% to 60% RH. The next desorption cycle shows a steady loss of about 3% by weight from 90% to 30% RH, then a sudden decrease to 0% by weight from 30% to 0% RH. The theoretical amount of water required for the formal monohydrate of the besylate salt is 3.6%. Thus, this version of the salt is hydrated to the trihydrate.

[0289] Besylate Pattern B - See Figure 48 During the first desorption cycle, the solid loses 1 wt.% from 50% RH to 0% RH. During the subsequent sorption cycles, the solid gains about 2.25% water up to 90% RH, but at a steady rate. This Pattern B version of the besylate salt shows a more favorable GVS profile than that of Pattern A.

[0290] Besylate Pattern C - See Figure 49 During the first desorption cycle, the solid loses about 3 wt.% from 50% RH to 0% RH. During the subsequent sorption cycles, the solid gains 3.75% water up to 90% RH, but there is a sharp increase of about 2.5 wt.% between 0% and 20% RH.

[0291] Naphthalene-2-sulfonate Pattern A - See Figure 50 During the first desorption cycle, the solid loses 1 wt.% from 50% RH to 0% RH. During the subsequent sorption cycles, the solid gains approximately 2.25% water up to 90% RH, but at a steady rate. XRPD analysis showed that there was no change in the crystallinity of the solid at the humidity extremes.

[0292] Malonate Pattern B - See Figure 51 The GVS profile shows that the material loses 5 wt% during the first desorption step to 0% RH. The material is therefore considered hygroscopic and was hydrated to a non-stoichiometric level at ambient conditions. During the subsequent sorption cycles, the solid gains 7.5% water up to 90% RH, but a sharp increase of about 3 wt% is observed between 30% and 40% RH. This water uptake is reversible, with absorbed water being lost with decreasing relative humidity. The theoretical amount of water required for the formal monohydrate of the malonate is 3.4%, so the salt is hydrated up to a dihydrate level at extreme conditions of humidity.

[0293] Example 4 (Further investigation of maleate salts) Five crystal patterns were identified for the maleate salt and designated Pattern A, Pattern B, Pattern C, Pattern D, and Pattern E. Data characterizing Patterns A, B, and C are provided above, and data characterizing Patterns D and E are provided below.

[0294] A comparison of the XRPD spectra of the above five crystal patterns and a mixture of A / B patterns is shown in FIG.

[0295] Patterns A, B, C, and D are believed to be variants with different degrees of hydration. Pattern A has proven difficult to isolate as it quickly transforms into a mixture of A and B upon absorption of water. Pattern B is a relatively stable hydrate, while Pattern C is believed to be a non-stoichiometric hydrate. Pattern D is also believed to be a non-stoichiometric hydrate and is similar to Pattern C. Pattern E is an N-methylpyrrolidone (NMP) solvate.

[0296] 4A. Preparation of Maleate Pattern A via Pattern A / B Mixture and Subsequent Thermal Cycling The free base of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile (4.9756 g) was charged to a round bottom flask and THF (204 mL, 41 volumes) was added. The mixture was heated to 60° C. To this solution was then added maleic acid (1.48 g, 1 equiv.) as a solution in THF (2 volumes). The mixture was left to equilibrate at 60° C. for 1 hour and then cooled to 20° C. overnight to give a beige suspension. The solid was isolated by vacuum filtration and washed with THF. The solid was dried under vacuum at 40° C. for 20 hours to give the maleate salt (SSA203).

[0297] A portion of the resulting solid (SSA203) was weighed (50 mg) into a crystallization tube and methyl isobutyl ketone (5 times the amount) was added. The mixture was equilibrated at room temperature for 18 hours to obtain a Pattern A / B mixture. A thermal cycle was performed by heating to 150° C. to obtain the maleate salt Pattern A.

[0298] (4B. Preparation of Maleate Pattern A by High Boiling Point Non-Aqueous Solvent Method) SSA203 (from Example 4A) was weighed into crystallization tubes (60 mg / tube) and the appropriate high boiling point solvent (10 volumes) was added. The mixture was equilibrated at RT for approximately 30 minutes, heated to 95° C., equilibrated for 4 hours, and then allowed to cool to RT over 70 hours. The mixture was then heated again to 95° C., equilibrated for 4 hours, and allowed to cool to RT over 3 hours. The solid was isolated and dried at 45° C. for 18 hours.

[0299] The solvents and the resulting maleate patterns are given in the table below.

[0300] [Table 16]

[0301] 4C. Antisolvent-Mediated Recrystallization 5-[[5-[4-(4-fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile maleate (SSA203 from Example 4A)) was weighed into two crystallization tubes and DMSO (4 volumes) or NMP (4 volumes) was added. The mixture was heated to 60°C. The yellow solution was then clarified and placed into a pre-heated 60°C clean tube. The clarified solution was then split into 320 μl aliquots such that each tube contained 80 mg of maleate. The solution was then added with the appropriate anti-solvent in 0.5-1 volume aliquots and allowed to equilibrate for a minimum of 10 minutes after each addition. This was done until a cloudy solution was formed or until 10 volumes of anti-solvent had been added. The mixture was then left to equilibrate at 60° C. for approximately 30 minutes, after which it was cooled to 25° C. and allowed to equilibrate for approximately 20 hours.

[0302] The entries that remained as a solution were cooled to 0° C. and allowed to equilibrate for approximately 6 hours. The mixture that remained as a solution at 0° C. was heated to 60° C. and approximately half of the solvent was evaporated by a gentle stream of nitrogen and cooled back to ambient temperature.

[0303] Crystal patterns isolated from various solvent combinations are shown below. Pattern B of solids isolated from DMSO / water, NMP / MeCN, and NMP / water Pattern A / B mixture from NMP / BuOH Pattern C isolated from DMSO / BuOH and DMSO / MeCN Pattern D isolated from THF+ flash evaporation Pattern E isolated from NMP / dioxane, NMP / n-PrOAc, NMP / toluene, NMP / THF, and NMP / EtOAc

[0304] The DSC and TGA profiles of the maleate salt pattern E are shown in FIG.

[0305] 4D. Preparation of Maleate Pattern D The free base of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile (4.9756 g) was charged to a round bottom flask and THF (204 mL, 41 volumes) was added. The mixture was heated to 60° C. Then, maleic acid (1.48 g, 1 equiv.) was added as a solution in THF (2 volumes) to the solution. The mixture was left to equilibrate at 60° C. for 1 hour. 100 ml of this solution was clarified and charged to a preheated 60° C. clean flask, allowed to cool to approximately 50° C., and flash evaporated to give maleate salt pattern D.

[0306] The DSC and TGA profiles of the maleate salt pattern E are shown in FIG.

[0307] 4E. Synthesis of amorphous maleate salt 5-[[5-[4-(4-fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile maleate (583.4 mg) was dissolved in hexafluoro-2-propanol (6F-IPA, 6 volumes, 1750 μL) at 30° C. The solution was clarified and placed in a tube containing tert-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 isolated by vacuum filtration and dried at 45° C. for 18 hours.

[0308] (4F. Formation of Maleic Acid Pattern B by Conditioning Pattern A) The maleate Pattern A was conditioned for 48 hours using a warm vacuum oven (25°C, with a slight vacuum bleed to activate flow in the oven) and a moisture source (static, tray of deionized water) with continuous monitoring by a multi-sample approach (XRPD samples) across the conditioning tray until all samples were reported as Pattern B.

[0309] (4G. Dynamic Vapor Sorption (DVS) Analysis of Maleate Pattern B) A defined amount of maleate salt Pattern B was placed in a pre-tared mesh stainless steel basket under ambient conditions. A complete experimental cycle consisted of five scans (desorption, repeated sorption, and desorption) at 10% RH intervals (60 min for each humidity level) over the range 0-90% at constant temperature (25 °C). Extended experiments of this kind should demonstrate the ability of the studied sample to absorb (or not absorb) moisture over a series of well-determined humidity ranges.

[0310] After cycling, the material was isolated at 0% RH and tested for crystallinity, then held at 90% RH for a minimum of 3 hours and retested for any change in crystallinity.

[0311] The results are shown in Figure 53.

[0312] The solid showed a moisture content of about 2.8 wt% before the first desorption. During the first sorption, the main weight gain occurred between 20% and 30% RH (about 2 wt%). After 5 cycles, the material returned to 0 and contained no moisture.

[0313] XRPD analysis showed a mixed phase at 0% RH and pattern B at 90% RH. This profile with hysteresis between 30% RH and 0% RH is typical of a reversible channel hydrate where the anhydrate to hydrate transition kinetically requires time to reach equilibrium above 30% RH.

[0314] (biological activity) Example A (Chk-1 kinase inhibitory activity) The compound of formula (1) (5-[[5-[4-(4-fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile) was tested for activity against Chk-1 kinase using the materials and protocol described below.

[0315] Reaction buffer: Base 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 * Required cofactors are added separately to each kinase reaction.

[0316] Reaction Procedure: (i) Prepare the indicated substrate in freshly prepared base reaction buffer. (ii) providing necessary cofactors to the substrate solution; (iii) The indicated kinase is added to the substrate solution and mixed gently. (iv) Compounds in DMSO are provided to the kinase reaction mixture. (v) 33P-ATP (specific activity 0.01 μCi / μl final) is supplied to the reaction mixture to initiate the reaction. (vi) The kinase reaction is incubated at room temperature for 120 minutes. (vii) The reactions are spotted onto P81 ion exchange filter paper (Whatman #3698-915). (viii) Wash the filter paper thoroughly in 0.1% phosphoric acid. (ix) The filter paper is dried and the counts are measured in a scintillation counter.

[0317] Kinase Information: CHK-1: Genbank accession number AF016582 Recombinant full-length construct, N-terminally GST-tagged, purified from insect cells. No special measures were taken to activate this kinase. Final concentration in assay = 0.5 nM Substrate:CHKtide Peptide sequence: [KKKVSRSGLYRSPSMPENLNRPR] Final concentration in assay = 20 μM No additional cofactors are added to the reaction mixture.

[0318] The results obtained according to the above protocol showed that the IC value of the compound of formula (1) against Chk-1 kinase was 50 The value was determined to be 0.00015 μM.

[0319] Example B (Gemcitabine combination cell assay) MIA PaCa-2 (ATCC CRL-1420) cells in logarithmic growth phase are trypsinized to detach the cells from the plate surface. Approximately 10,000 cells / well are seeded into 96-well plates in RPMI containing 10% fetal bovine serum, 1% sodium pyruvate, and 1% L-GlutaMax. Cells are allowed to adhere to the plate surface overnight. 10% of each Chk1 inhibitor test compound and gemcitabine are added to the plate. 0.5A two-fold dilution series is made to final top concentrations of 3000 nM and 100 nM, respectively. Each Chk1 inhibitor is combined with gemcitabine such that each concentration of Chk1 inhibitor is added to each concentration of gemcitabine. Each agent is also tested as a single agent. Agents are added (in duplicate) to adherent cells and incubated for 72 hours. At 72 hours, cells are treated with Promega Cell Titer Glo reagent for approximately 15 minutes. Luminescence (relative light units, RLU) is recorded using a BMG Polarstar Omega plate reader. The single agent concentration that reduces the total signal by 50% (IC 50 ) is calculated using PRISM software and four-parameter nonlinear regression curve fitting. For combination studies, RLUs are plotted on an XY plot using PRISM with gemcitabine concentration on the X-axis and RLU on the Y-axis. RLUs per concentration of Chk1 inhibitor are plotted as a function of gemcitabine concentration. The IC50 of gemcitabine alone at each concentration of Chk1 is determined using four-parameter nonlinear regression curve fitting. Gemcitabine Alone IC 50 The approximate concentrations of Chk1 inhibitors that result in a 2-fold and 10-fold decrease in are calculated as an index of synergistic potency.

[0320] From the results obtained according to the above protocol, the IC value of the compound of formula (1) alone against MIAPaca-2 cells was 50 Value(Chk1 IC 50 ) and the IC of gemcitabine alone for the compound of formula (1) 50 Approximate compound concentrations that reduce by a factor of two (2xLS) and by a factor of ten (10xLS) are shown below.

[0321] [Table 17]

[0322] (Pharmaceutical preparations) (i) Tablet formulation A tablet composition containing any one of embodiments 1.1 to 1.48 or a pharma- ceutically acceptable salt as defined in the Examples 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 to form a tablet by known methods.

[0323] (ii) Capsule formulation A capsule formulation is prepared by mixing 100 mg of a pharma- ceutically acceptable salt as defined in any one of embodiments 1.1 to 1.48 or in the examples above with 100 mg of lactose and filling the resulting mixture into a standard opaque hard gelatin capsule.

[0324] (iii) Injection preparation I A parenteral composition for administration by injection can be prepared by dissolving any one of the pharma- ceutically acceptable salts according to any one of the embodiments 1.1 to 1.48 or the above examples in water containing 10% propylene glycol to give a concentration of 1.5% by weight of the active compound, which is then sterilized by filtration, filled into an ampoule and sealed.

[0325] (iv) Injection Preparation II A parenteral composition for injection is prepared by dissolving a pharma- ceutically acceptable salt as defined in any one of embodiments 1.1 to 1.48 or in the above examples (2 mg / ml) and mannitol (50 mg / ml) in water, sterile filtering the solution and filling into sealable 1 ml vials or ampoules.

[0326] (v) Injection preparation III A formulation for intravenous delivery by injection or infusion can be prepared by dissolving any one of the pharma- ceutically acceptable salts as defined in any one of the embodiments 1.1 to 1.48 or in the above examples in water at 20 mg / ml. The vial is then sealed and sterilized by autoclaving.

[0327] (vi) Injection preparation IV A formulation for intravenous delivery by injection or infusion can be prepared by dissolving any one of the pharma- ceutically acceptable salts of embodiment 1.1 to embodiment 1.48 or as defined in the above examples in water containing a buffer (e.g., 0.2 M acetate, pH 4.6) at 20 mg / ml. The vial is then sealed and sterilized by autoclaving.

[0328] (vii) Subcutaneous injection formulation A composition for subcutaneous administration is prepared by mixing a pharma- ceutically acceptable salt as defined in any one of embodiments 1.1 to 1.48 or in the above examples with pharmaceutical grade corn oil to a concentration of 5 mg / ml.The composition is sterilized and filled into a suitable container.

[0329] (viii) Lyophilized preparation Aliquots of the formulated pharma- ceutically acceptable salts of any one of embodiment 1.1 to embodiment 1.48 or as defined in the examples above are placed in 50 ml vials and lyophilized. During lyophilization, the compositions are frozen at (-45°C) using a one-step freezing protocol. The temperature is increased to -10°C for annealing, then decreased to freezing at -45°C, followed by primary drying at +25°C for about 3400 minutes, followed by secondary drying at stepwise increasing temperatures to 50°C. The pressure during primary and secondary drying is set at 80 mTorr.

[0330] (Equivalent) The above examples are presented for the purpose of illustrating the present invention, and are not intended to limit the scope of the present invention in any way. It will be readily apparent that many modifications and variations may be made to the specific embodiments of the present invention as described and illustrated above, without departing from the principles underlying the invention. All such modifications and variations are intended to be embraced by this application.

Claims

1. A pharma- ceutically acceptable salt of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile selected from maleate, tosylate, besylate, and malonate salts.

2. The pharma- ceutically acceptable salt of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile of claim 1 which is the maleate salt.

3. 2. The pharma- ceutically acceptable salt of claim 1, wherein the salt ratio (molar ratio of acid:free base) is about 1:

1.

4. 2. The pharma- ceutically acceptable salt of claim 1, having a crystallinity of from 50% to 100%.

5. 3. The pharma- ceutically acceptable maleic acid Pattern B salt of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile according to claim 2, having an XRPD spectrum characterized by major °2Th (°2Theta) peaks at 6.9±0.2° and / or 26.4±0.2° and / or 11.8±0.2° and / or 17.9±0.2°.

6. 23. The pharma- ceutically acceptable maleate salt of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile according to claim 2, which is a Pattern B salt having an XRPD spectrum substantially as shown in Figure 25.

7. A pharmaceutical composition comprising a pharma- ceutically acceptable salt of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile as defined in any one of claims 1 to 6, and a pharma- ceutically acceptable excipient.

8. A pharmaceutical composition for treating cancer comprising a pharma- ceutically acceptable salt of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile as defined in any one of claims 1 to 6.

9. A pharmaceutical combination comprising a pharma- ceutically acceptable salt of 5-[[5-[4-(4-fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile as defined in any one of claims 1 to 6 and another therapeutically active agent.

10. 7. A process for preparing a pharma- ceutically acceptable salt as defined in any one of claims 1 to 6, comprising dispersing 5-[[5-[4-(4-fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile in tetrahydrofuran to form a mixture, heating the mixture to an elevated temperature in the range of 45°C to 65°C, e.g. 55°C to 65°C, in particular about 60°C, adding the requisite amount of acid to the mixture, maintaining the mixture at or near the elevated temperature for a defined period of time, and cooling the mixture to isolate the pharma- ceutically acceptable salt.

11. 7. A process for preparing a pharma- ceutically acceptable salt as defined in any one of claims 1 to 6, comprising dispersing 5-[[5-[4-(4-fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile in a mixture of tetrahydrofuran and acetonitrile (e.g., a 1:1 mixture) to form a mixture, heating the mixture to an elevated temperature in the range of 45°C to 55°C (e.g., about 50°C), adding the requisite amount of acid to the mixture, maintaining the mixture at or near the elevated temperature for a defined period of time, and cooling the mixture to isolate the pharma- ceutically acceptable salt.

12. 7. A process for the preparation of a pharma- ceutically acceptable salt as defined in any one of claims 1 to 6, comprising dispersing 5-[[5-[4-(4-fluoro-1-methyl-4-piperidyl)-2-methoxy-phenyl]-1H-pyrazol-3-yl]amino]pyrazine-2-carbonitrile in a mixture of tetrahydrofuran and water (e.g., the mixture contains 75% (v / v) to 97% (v / v) tetrahydrofuran and 3% (v / v) to 25% (v / v) water, more preferably about 95% (v / v) tetrahydrofuran and about 5% (v / v) water) to form a mixture, heating the mixture to an elevated temperature in the range of 45°C to 65°C (e.g., about 50°C to 60°C), adding the required amount of acid to the mixture, maintaining the mixture at or near the elevated temperature for a defined period of time, and cooling the mixture to isolate the pharma- ceutically acceptable salt.

13. 11. The method of claim 10, wherein the acid is maleic acid and the resulting pharma- ceutically acceptable salt is a maleate salt.

14. 14. The method of claim 13, wherein the maleate salt is a maleate Pattern A salt.

15. 15. The method of claim 14, further comprising converting the Pattern A maleate to a Pattern B maleate by conditioning the Pattern A salt in an atmosphere having a relative humidity of greater than 50%.

16. The invention as defined in any one of embodiments 1.1 to 1.48, embodiments 2.1 to 2.16, embodiments 3.1 to 3.30, embodiment 4.1, and embodiment 5.1 to embodiment 5.13 herein.

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