Salts and solid state forms of KIF18A inhibitor compounds

Abstract

Disclosed herein are salts, crystalline anhydrous forms, hydrates, solvates or co-crystals of the free base compound 2-(6-azaspiro[2.5]octan-6-yl)-N-[2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl]-4-[(2-hydroxyethanesulfonyl)amino]benzamide (Compound A); methods of preparation, pharmaceutical compositions, and methods of treating a disease mediated by inhibition of the motor protein kinesin family member 18A (KIF18A), wherein the disease is a neoplastic disease including cancer or tumor.

Description

[Technical field]

[0001] Incorporation by Reference of Electronically Submitted Materials This specification incorporates by reference in its entirety the ST.26 format computer readable nucleotide / amino acid sequence listing, which was submitted contemporaneously herewith and is identified as follows: A-2832-WO01-SEC_FromUS-PSP_Seq_Listing_ST26_072122b, a 137 KB XML format file created on July 21, 2022.

[0002] The present disclosure relates to a salt, hydrate, solvate or co-crystal of the free base compound 2-(6-azaspiro[2.5]octan-6-yl)-N-[2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl]-4-[(2-hydroxyethanesulfonyl)amino]benzamide (Compound A); or a solid form of Compound A, including a crystalline anhydrous form, a salt, hydrate, solvate or co-crystal thereof; methods of preparation, pharmaceutical compositions, and methods of treating diseases mediated by inhibition of the motor protein kinesin family member 18A (KIF18A). [Background technology]

[0003] The free base compound 2-(6-azaspiro[2.5]octan-6-yl)-N-[2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl]-4-[(2-hydroxyethanesulfonyl)amino]benzamide (Compound A) is useful as an inhibitor of the motor protein kinesin family member 18A (KIF18A). [ka]

[0004] Kinesins are molecular motors that play an important role in cell division and the transport of intracellular vesicles and organelles. Mitotic kinesins play a role in several aspects of spindle assembly, chromosome segregation, centrosome separation, and dynamics. Human kinesins are classified into 14 subfamilies based on sequence homology within the so-called "motor domain." The ATPase activity of this domain is the driving force for unidirectional movement along microtubules (MTs). The non-motor domains of these proteins are responsible for cargo binding. The "cargo" may include any one of a variety of membranous organelles, signaling scaffolding systems, and chromosomes. Kinesins use the energy of ATP hydrolysis to move cargo along polarized microtubules. Thus, kinesins are often referred to as "plus-end" or "minus-end" directional motors.

[0005] The KIF18A gene belongs to the kinesin-8 subfamily and is a plus-end directed motor. KIF18A is thought to affect the dynamics of the plus ends of kinetochore microtubules, controlling correct chromosome positioning and spindle tension. Deletion of human KIF18A results in longer spindles, increased chromosome oscillations at metaphase, and activation of the mitotic spindle assembly checkpoint in HeLa cervical cancer cells. KIF18A appears to be a viable target for cancer therapy. KIF18A is overexpressed in a variety of cancers, including but not limited to colon, breast, lung, pancreatic, prostate, bladder, head, neck, cervical, and ovarian cancers. Furthermore, genetic deletion or knockdown or inhibition of KIF18A affects the mitotic spindle apparatus in cancer cell lines. In particular, inhibition of KIF18A has been found to induce mitotic cell arrest, mitotic catastrophe, or multipolarity-driven lethality or death after mitotic slippage in interphase, known vulnerabilities that can promote cell death during mitosis via apoptosis.

[0006] The human KIF18A gene sequence, the human KIF18A mRNA sequence, and the encoded KIF18A protein are provided herein as SEQ ID NOs: 12, 13 and 11, respectively.

[0007] Compound A, as well as representative methods for producing it, are described in International Patent Application Publication No. WO2020 / 132648, the entire contents of which are incorporated herein by reference. However, stable salts, hydrates, solvates, or cocrystals of Compound A are desirable, particularly for commercial pharmaceutical production of Compound A, along with solid forms of Compound A (including crystalline anhydrous Compound A or amorphous Compound A). Summary of the Invention

[0008] In one embodiment, disclosed herein is a salt, hydrate, solvate, or co-crystal of Compound A having the structure shown below: [ka] (the chemical name is 2-(6-azaspiro[2.5]octan-6-yl)-N-[2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl]-4-[(2-hydroxyethanesulfonyl)amino]benzamide, or also known as N-(2-(4,4-difluoropiperidin-1-yl)-6-methylpyrimidin-4-yl)-4-((2-hydroxyethyl)sulfonamido)-2-(6-azaspiro[2.5]octan-6-yl)benzamide), or a solid form of Compound A (including crystalline anhydrous Compound A or amorphous Compound A), or a salt, hydrate, solvate, or co-crystal thereof.

[0009] In another embodiment, disclosed herein are solid forms of Compound A, including crystalline anhydrous forms, salts, hydrates, solvates, or co-crystals of Compound A. The solid forms can be crystalline or amorphous forms.

[0010] In various embodiments, disclosed herein are compounds including Compound A hydrochloride (Compound A-HCl), mesylate (Compound A-MsA), tosylate (Compound A-TsA), sulfate (Compound A-Sulfate), variable hydrate (Compound A-variable hydrate), tetrahydrofuran solvate (Compound A-THF), ethanol solvate (Compound A-Ethanol), 1-propanol solvate (Compound A-1-Propanol), isopropyl alcohol solvate (Compound A-IPA), methanol solvate (Compound A-Methanol), isopropyl acetate solvate (Compound A-IPAc), acetone solvate (Compound A-Acetone), cyclopentyl methyl ether solvate (Compound A-CPME), dioxane solvate (Compound A-Dioxane), ethyl acetate solvate (Compound A-EtOAc), 2. The salt, anhydrate, hydrate, solvate, or co-crystal of claim 1 selected from an acetonitrile solvate (Compound A-MeCN), a methyl tert-butyl ether solvate (Compound A-MTBE) a toluene solvate (Compound A-toluene), a dodecyl sulfate salt (Compound A-dodecyl sulfate), a dimethylformamide (DMF) solvate hydrate (Compound A-DMF-hydrate), a dimethylacetamide (DMAC) solvate (Compound A-DMAC), a monobesylate salt hydrate (Compound A-besylate-hydrate), a caffeine co-crystal (Compound A-caffeine), a citric acid co-crystal (Compound A-citric acid), a saccharin co-crystal (Compound A-saccharin), an L-tartaric acid co-crystal (Compound A-AL-tartaric acid), or a urea co-crystal (Compound A-urea); or a solid form thereof.

[0011] In embodiment 1, the present invention provides a hydrochloride salt of Compound A, having the structure shown below: [ka]

[0012] In embodiment 1a, the present invention provides a solid form of Compound A-HCl. In a subembodiment, the solid form is crystalline Form 1 (Compound A-HCl-Form 1). In another subembodiment, the solid form is crystalline Form 2 (Compound A-HCl-Form 2).

[0013] In embodiment 1b, the present invention relates to a solid phase 19 Provided is crystalline Compound A-HCl-Form 1, characterized by F NMR peaks at -91 and -103 ppm.

[0014] In embodiment 1c, the present invention provides crystalline Compound A-HCl-Form 1, further characterized by X-ray powder diffraction (XRPD) pattern peaks 7.5, 16.9, and 20.2±0.2 degrees 2θ using CuKα radiation.

[0015] In embodiment 1d, the present invention provides crystalline Compound A-HCl-Form 1, further characterized by X-ray powder diffraction (XRPD) pattern peaks of 12.8, 18.2, 22.7, 23.6, 24.8, and 26.1±0.2 degrees 2θ using CuKα radiation.

[0016] In embodiment 1e, the present invention provides crystalline Compound A-HCl-Form 1, further characterized by X-ray powder diffraction (XRPD) pattern peaks of 10.9, 14.5, 15.7, 15.9, 19.8, 20.6, 21.6, 23.2, 26.1, and 26.8±0.2 degrees 2θ using CuKα radiation.

[0017] In embodiment 1f, the present invention provides crystalline Compound A-HCl-Form 1, having an XRPD pattern substantially as shown in FIG.

[0018] In embodiment 1g, the present invention provides crystalline Compound A-HCl-Form 1, having an endothermic transition at 268.5° C. to 274.5° C. as measured by differential scanning calorimetry.

[0019] In embodiment 1h, the present invention provides crystalline Compound A-HCl-Form 1, wherein the endothermic transition is 271.5°C ± 3°C.

[0020] In embodiment 1i, the present invention provides crystalline Compound A-HCl-Form 1, having a thermogravimetric analysis (TGA) substantially as shown in FIG.

[0021] In embodiment 1j, the present invention provides crystalline Compound A-HCl-Form 1, having a single crystal structure substantially as shown in FIG.

[0022] In embodiment 2, the present invention provides a mesylate salt of Compound A, having the structure shown below: [ka]

[0023] In embodiment 2a, the invention provides a solid form of Compound A-MsA. In a subembodiment, the solid form is crystalline Form 1 (Compound A-MsA-Form 1). In another subembodiment, the solid form is crystalline Form 2 (Compound A-MsA-Form 2).

[0024] In embodiment 2b, the present invention relates to a solid phase 19 Provided is a crystalline compound A-MsA-Form 1, characterized by F NMR peaks at -95.2 and -103.2±0.5 ppm. Spinning side bands are indicated with (*).

[0025] In embodiment 2c, the present invention provides crystalline compound A-MsA-Form 1, further characterized by X-ray powder diffraction (XRPD) pattern peaks of 7.0, 16.5, and 23.9±0.2 degrees 2θ using CuKα radiation.

[0026] In embodiment 2d, the present invention provides the crystalline compound A-MsA-Form 1, further characterized by XRPD pattern peaks 12.6, 15.7, 17.4, 18.5, 20.0 and 21.0±0.2 degrees 2θ using CuKα radiation.

[0027] In embodiment 2e, the present invention provides the crystalline compound A-MsA-Form 1, further characterized by XRPD pattern peaks 5.8, 11.8, 13.5, 15.3, 16.1, 18.0, 20.6, 25.2, 28.0 and 30.5±0.2 degrees 2θ using CuKα radiation.

[0028] In embodiment 2f, the present invention provides a crystalline compound A-MsA-Form 1, having an XRPD pattern substantially as shown in FIG.

[0029] In embodiment 2g, the present invention provides crystalline Compound A-MsA-Form 1, having an endothermic transition at 247° C. to 253° C. as measured by differential scanning calorimetry.

[0030] In embodiment 2h, the present invention provides crystalline compound A-MsA-Form 1, which has an endothermic transition at 250°C ± 3°C.

[0031] In embodiment 2i, the present invention provides a crystalline compound A-MsA-Form 1 having a thermogravimetric analysis (TGA) substantially as shown in FIG.

[0032] In embodiment 3, the present invention provides a tosylate salt of Compound A, having the structure shown below: [ka]

[0033] In embodiment 3a, the invention provides a solid form of Compound A-TsA. In a subembodiment, the solid form is crystalline form 1 (Compound A-TsA-Form 1). In another subembodiment, the solid form is crystalline form 2 (Compound A-TsA-Form 2). In a subembodiment, the solid form is crystalline form 3 (Compound A-TsA-Form 3). In another subembodiment, the solid form is crystalline form 4 (Compound A-TsA-Form 4). In another subembodiment, the solid form is crystalline form 5 (Compound A-TsA-Form 5). In yet another subembodiment, the solid form is ditosylate crystalline form 6 (Compound A-DiTsA-Form 6).

[0034] In embodiment 3b, the present invention provides a crystalline compound A-TsA-Form 4, characterized by X-ray powder diffraction (XRPD) pattern peaks 6.2, 14.7, and 23.5±0.2 degrees 2θ using CuKα radiation.

[0035] In embodiment 3c, the present invention provides crystalline compound A-TsA-Form 4, further characterized by XRPD pattern peaks 10.5, 12.4, 14.2, 19.1, 21.5 and 29.0±0.2 degrees 2θ using CuKα radiation.

[0036] In embodiment 3d, the present invention provides crystalline compound A-TsA-Form 4, further characterized by XRPD pattern peaks 15.5, 16.5, 17.7, 18.3, 18.6, 20.1, 20.8, 24.1, and 25.3±0.2 degrees 2θ using CuKα radiation.

[0037] In embodiment 3e, the present invention provides a crystalline compound A-TsA-Form 4 having an XRPD pattern substantially as shown in Figure 24a.

[0038] In embodiment 3f, the present invention provides a crystalline compound A-TsA-Form 4 having a single crystal structure substantially as shown in Figure 24b.

[0039] In embodiment 3g, the present invention provides crystalline Compound A-TsA-Form 4, which has an endothermic transition at 250° C. to 256° C. as measured by differential scanning calorimetry.

[0040] In embodiment 3h, the present invention provides crystalline compound A-TsA-Form 4, wherein the endothermic transition is at 253°C ± 3°C.

[0041] In embodiment 3i, the present invention provides a crystalline compound A-TsA-Form 4 having a thermogravimetric analysis (TGA) substantially as shown in FIG.

[0042] In embodiment 3j, the present invention provides a solid phase 19 Provided is a crystalline compound A-TsA-Form 4, characterized by F NMR peaks of -96.93 and -101.60 ± 0.5 ppm. Spinning side bands are indicated with (*).

[0043] In embodiment 4, the present invention provides a solid form of Compound A. In a subembodiment, the solid form is an amorphous form (Compound A-amorphous). In another subembodiment, the solid form is crystalline Compound A-Form 1 (Compound A-Form 1).

[0044] In embodiment 4a, the present invention provides Compound A-amorphous having an XRPD pattern substantially as shown in FIG.

[0045] In embodiment 4b, the invention provides Compound A-amorphous having a melting onset of 88° C. to 94° C. as measured by differential scanning calorimetry. In a subembodiment, Compound A-amorphous has a melting onset at 91° C.±3° C. In a subembodiment, Compound A-amorphous has a DSC thermograph pattern substantially as shown in FIG.

[0046] In embodiment 4c, the present invention provides Compound A-amorphous having a thermogravimetric analysis (TGA) substantially as shown in FIG.

[0047] In embodiment 4d, the present invention provides crystalline Compound A-Form 1 having a thermogravimetric analysis (TGA) substantially as shown in FIG.

[0048] In embodiment 5, the present invention provides a sulfate salt of Compound A having the structure: [ka]

[0049] In embodiment 5a, the invention provides a solid form of Compound A-sulfate. In a subembodiment, the solid form is crystalline Form 1 (Compound A-sulfate-Form 1). In another subembodiment, Compound A-sulfate-Form 1 has an XRPD pattern substantially as shown in FIG. 30. In another subembodiment, Compound A-sulfate-Form 1 has an endothermic transition at 261° C. to 267° C. as measured by differential scanning calorimetry. In yet another subembodiment, Compound A-sulfate-Form 1 has an endothermic transition at 264° C.±3° C. In yet another subembodiment, Compound A-sulfate-Form 1 has a thermogravimetric analysis (TGA) pattern substantially as shown in FIG. 31.

[0050] In embodiment 6, the present invention provides a hydrate of Compound A having the structure as shown below: [ka] (wherein n is a number ranging from 0.5 to 2, or a variable (mixture) thereof. The value of n may vary as a result of different preparation methods and / or storage conditions.)

[0051] In embodiment 6a, the present invention provides a solid form of Compound A-hydrate.

[0052] In embodiment 6b, the present invention provides Compound A-variable-hydrate-Form 2, characterized by X-ray powder diffraction (XRPD) pattern peaks of 13.9, 16.2, and 19.6±0.2 degrees 2θ using CuKα radiation.

[0053] In embodiment 6c, the present invention provides Compound A-variable-hydrate-Form 2, further characterized by XRPD pattern peaks 3.5, 17.4, 18.4, 18.7, 20.0, 20.2, 22.6, 22.9, 27.5, and 30.8±0.2 degrees 2θ using CuKα radiation.

[0054] In embodiment 6d, the present invention provides Compound A-variable-hydrate-Form 2, further characterized by XRPD pattern peaks 3.5, 10.1, 11.2, 13.9, 16.2, 18.2, 19.2, 23.2, and 26.0±0.2 degrees 2θ using CuKα radiation.

[0055] In embodiment 6e, the present invention provides Compound A-variable hydrate-Form 2 having an XRPD pattern substantially as shown in FIG.

[0056] In embodiment 6f, the present invention provides Compound A-variable-hydrate-form 2 having a dehydration onset at 48° C.-54° C. and a melting point of 136° C. as measured by differential scanning calorimetry. In a subembodiment, Compound A-variable-hydrate-form 2 has a DSC thermograph pattern substantially as shown in FIG.

[0057] In embodiment 6g, the present invention provides Compound A-variable-hydrate-form 2, which has an endothermic transition at 51°C ± 3°C.

[0058] In embodiment 6h, the present invention provides Compound A-variably-hydrated-Form 2 having a thermogravimetric analysis (TGA) substantially as shown in FIG.

[0059] In embodiment 7, the present invention provides a crystalline anhydrous form of Compound A (Compound A-Anhydrous).

[0060] In embodiment 7a, the solid form is crystalline anhydrous Form 3 (Compound A-Anhydrous-Form 3). In a subembodiment, Compound A-Anhydrous-Form 3 has an XRPD pattern substantially as shown in FIG. 40. In another subembodiment, Compound A-Anhydrous-Form 3 has an onset of melting at 193.5° C. to 199.5° C. as measured by differential scanning calorimetry. In yet another subembodiment, Compound A-Anhydrous-Form 3 has an onset of melting at 196.5° C.±3° C. In yet another subembodiment, Compound A-Anhydrous-Form 3 has a dynamic vapor sorption (DVS) substantially as shown in FIG. 42.

[0061] In embodiment 7b, the solid form is crystalline anhydrous Form 4 (Compound A-Anhydrous-Form 4). In a subembodiment, Compound A-Anhydrous-Form 4 has an XRPD pattern substantially as shown in FIG.

[0062] In embodiment 7c, the solid form is crystalline anhydrous Form 5 (Compound A-Anhydrous-Form 5). In a subembodiment, Compound A-Anhydrous-Form 5 has an XRPD pattern substantially as shown in FIG. 44. In another subembodiment, Compound A-Anhydrous-Form 5 has a melting onset at 188.5° C. to 194.5° C., substantially as shown in FIG. 45, as measured by differential scanning calorimetry. In yet another subembodiment, Compound A-Anhydrous-Form 5 has a melting onset at 191.5° C.±3° C. In yet another subembodiment, Compound A-Anhydrous-Form 5 has a dynamic vapor sorption (DVS) pattern substantially as shown in FIG. 46, indicating that Anhydrous Form 5 has rehydrated to Compound A-monohydrate.

[0063] In embodiment 7d, the solid form is crystalline anhydrous Form 6 (Compound A-Anhydrous-Form 6). In a subembodiment, Compound A-Anhydrous-Form 6 has an XRPD pattern substantially as shown in Figure 47. In another subembodiment, Compound A-Anhydrous-Form 6 has an onset of melting at 183.4°C to 189.4°C as measured by differential scanning calorimetry. In yet another subembodiment, Compound A-Anhydrous-Form 6 has an onset of melting at 186.4°C ± 3°C.

[0064] In embodiment 7e, the solid form is crystalline anhydrous Form 7 (Compound A-Anhydrous-Form 7). In a subembodiment, Compound A-Anhydrous-Form 7 has an XRPD pattern substantially as shown in FIG.

[0065] In embodiment 7f, the solid form is crystalline anhydrous Form 8 (Compound A-Anhydrous-Form 8). In a subembodiment, Compound A-Anhydrous-Form 8 has an XRPD pattern substantially as shown in FIG.

[0066] In embodiment 8, the present invention provides a tetrahydrofuran (THF) solvate of Compound A, having the structure shown below: [ka]

[0067] In embodiment 8a, the invention provides a solid form of Compound A-THF. In a subembodiment, Compound A-THF has an XRPD pattern substantially as shown in FIG. 53. In another subembodiment, Compound A-THF has an onset of melting at 188.5° C. to 194.5° C. as measured by differential scanning calorimetry. In yet another subembodiment, Compound A-THF has an onset of melting at 191.5° C.±3° C. In yet another subembodiment, Compound A-THF has a thermogravimetric analysis (TGA) substantially as shown in FIG. 54.

[0068] In embodiment 9, the present invention provides an ethanol solvate of Compound A. In embodiment 9a, the present invention provides a solid form of Compound A-ethanol. In a subembodiment, Compound A-ethanol has an XRPD pattern substantially as shown in FIG. 55. In another subembodiment, Compound A-ethanol has an onset of melting at 162.6° C. to 168.6° C. as measured by differential scanning calorimetry. In yet another subembodiment, Compound A-ethanol has an onset of melting at 165.6° C.±3° C. In yet another subembodiment, Compound A-ethanol has a thermogravimetric analysis (TGA) substantially as shown in FIG. 56.

[0069] In embodiment 10, the present invention provides a 1-propanol solvate (Compound A-1-propanol). In embodiment 10a, the present invention provides a solid form of Compound A-1-propanol. In a subembodiment, Compound A-1-propanol has an XRPD pattern substantially as shown in FIG. 58. In another subembodiment, Compound A-1-propanol has an onset of melting at 191.2° C. to 197.2° C. as measured by differential scanning calorimetry. In yet another subembodiment, Compound A-1-propanol has an onset of melting at 194.2° C.±3° C. In yet another subembodiment, Compound A-1-propanol has a thermogravimetric analysis (TGA) substantially as shown in FIG. 59.

[0070] In embodiment 11, the present invention provides an isopropyl alcohol solvate of Compound A (Compound A-IPA). In embodiment 11a, the present invention provides a solid form of Compound A-IPA. In a subembodiment, Compound A-IPA has an XRPD pattern substantially as shown in FIG. 60. In another subembodiment, Compound A-IPA has an onset of melting at 155.7° C. to 161.7° C. as measured by differential scanning calorimetry. In yet another subembodiment, Compound A-IPA has an onset of melting at 158.7° C.±3° C. In yet another subembodiment, Compound A-IPA has a thermogravimetric analysis (TGA) substantially as shown in FIG. 61.

[0071] In embodiment 12, the present invention provides a methanol solvate of Compound A (Compound A-methanol). In embodiment 12a, the present invention provides a solid form of Compound A-methanol. In a subembodiment, Compound A-methanol has an XRPD pattern substantially as shown in Figure 62.

[0072] In embodiment 13, the present invention provides an isopropyl acetate solvate of Compound A (Compound A-IPAc). In embodiment 13a, the present invention provides a solid form of Compound A-IPAc. In a subembodiment, Compound A-IPAc has an XRPD pattern substantially as shown in Figure 63.

[0073] In embodiment 14, the present invention provides an acetone solvate of Compound A (Compound A-acetone). In embodiment 14a, the present invention provides a solid form of Compound A-acetone. In a subembodiment, Compound A-acetone has an XRPD pattern substantially as shown in Figure 64.

[0074] In embodiment 15, the present invention provides a cyclopentyl methyl ether solvate of Compound A (Compound A-CPME). In embodiment 15a, the present invention provides a solid form of Compound A-CPME. In a subembodiment, Compound A-CPME has an XRPD pattern substantially as shown in Figure 65.

[0075] In embodiment 16, the present invention provides a dioxane solvate of Compound A (Compound A-dioxane). In embodiment 16a, the present invention provides a solid form of Compound A-dioxane. In a subembodiment, Compound A-dioxane has an XRPD pattern substantially as shown in Figure 66.

[0076] In embodiment 17, the present invention provides an ethyl acetate solvate of Compound A (Compound A-EtOAc). In embodiment 17a, the present invention provides a solid form of Compound A-EtOAc. In a subembodiment, Compound A-EtOAc has an XRPD pattern substantially as shown in Figure 67.

[0077] In embodiment 18, the present invention provides an acetonitrile solvate of Compound A (Compound A-MeCN). In embodiment 18a, the present invention provides a solid form of Compound A-MeCN. In a subembodiment, Compound A-MeCN has an XRPD pattern substantially as shown in Figure 68.

[0078] In embodiment 19, the present invention provides a methyl tert-butyl ether solvate of Compound A (Compound A-MTBE). In embodiment 19a, the present invention provides a solid form of Compound A-MTBE. In a subembodiment, Compound A-MTBE has an XRPD pattern substantially as shown in Figure 69.

[0079] In embodiment 20, the present invention provides a toluene solvate of Compound A (Compound A-toluene). In embodiment 20a, the present invention provides a solid form of Compound A-toluene. In a subembodiment, Compound A-toluene has an XRPD pattern substantially as shown in Figure 70.

[0080] In embodiment 21, the present invention provides a dodecyl sulfate salt of Compound A (Compound A-dodecyl sulfate). In embodiment 21a, the present invention provides a solid form of dodecyl sulfate salt (Compound A-dodecyl sulfate). In a subembodiment, Compound A-dodecyl sulfate has an XRPD pattern substantially as shown in Figure 71.

[0081] In embodiment 22, the present invention provides a dimethylformamide (DMF) solvate hydrate of Compound A (Compound A-DMF-hydrate). In embodiment 22a, the present invention provides a solid form of Compound A-DMF-hydrate. In a subembodiment, Compound A-DMF-hydrate has an XRPD pattern substantially as shown in FIG. 73. In another subembodiment, Compound A-DMF-hydrate has an onset melting temperature of 104.8° C. to 110.8° C. as measured by differential scanning calorimetry. In yet another subembodiment, Compound A-DMF-hydrate has an onset melting temperature of 107.8° C.±3° C. In yet another subembodiment, Compound A-DMF-hydrate has a DSC pattern substantially as shown in FIG. 74.

[0082] In embodiment 23, the invention provides a dimethylacetamide (DMAC) solvate of Compound A (Compound A-DMAC). In embodiment 23a, the invention provides a solid form of Compound A-DMAC. In a subembodiment, Compound A-DMAC has an XRPD pattern substantially as shown in FIG. 75. In another subembodiment, Compound A-DMAC has an onset of melting at 147° C. to 153° C. as measured by differential scanning calorimetry. In yet another subembodiment, Compound A-DMAC has an onset of melting at 150° C.±3° C. In yet another subembodiment, Compound A-DMAC has a DSC pattern substantially as shown in FIG. 76.

[0083] In embodiment 24, the present invention provides a monobesylate hydrate of Compound A (Compound A-besylate-hydrate). In embodiment 24a, the present invention provides a solid form of Compound A-besylate-hydrate. In embodiment 24b, the present invention provides a solid form of Compound A-besylate-hydrate Form 1. In a subembodiment, Compound A-besylate-hydrate Form 1 has an XRPD pattern substantially as shown in Figure 77. In yet another subembodiment, Compound A-besylate-hydrate Form 1 has a DSC pattern substantially as shown in Figure 78.

[0084] In embodiment 25, the present invention provides a caffeine cocrystal of Compound A (Compound A-caffeine). In embodiment 25a, the present invention provides a solid form of Compound A-caffeine. In embodiment 25b, the solid form of Compound A-caffeine is crystalline Compound A-caffeine cocrystal Form 1. In a subembodiment, Compound A-caffeine cocrystal Form 1 has an XRPD pattern substantially as shown in Figure 79. In yet another subembodiment, Compound A-caffeine cocrystal Form 1 has a DSC pattern substantially as shown in Figure 80. In yet another subembodiment, Compound A-caffeine cocrystal Form 1 has a DVS pattern substantially as shown in Figure 81.

[0085] In embodiment 26, the invention provides a citric acid co-crystal of Compound A (Compound A-citric acid). In embodiment 26a, the invention provides a solid form of Compound A-citric acid. In embodiment 26b, the solid form of Compound A-citric acid is crystalline Compound A-citric acid co-crystal Form 1. In a subembodiment, Compound A-citric acid co-crystal Form 1 has an XRPD pattern substantially as shown in Figure 82. In yet another subembodiment, Compound A-citric acid co-crystal Form 1 has a DSC pattern substantially as shown in Figure 83.

[0086] In embodiment 26c, the solid form of Compound A-citric acid is crystalline Compound A citric acid co-crystal Form 2. In a subembodiment, Compound A-citric acid co-crystal Form 2 has an XRPD pattern substantially as shown in Figure 84. In yet another subembodiment, Compound A-citric acid co-crystal Form 2 has DSC and TGA patterns substantially as shown in Figure 85.

[0087] In embodiment 27, the present invention provides a saccharin cocrystal of Compound A (Compound A-saccharin). In embodiment 27a, the present invention provides a solid form of Compound A-saccharin. In embodiment 27b, the solid form of Compound A-saccharin is crystalline Compound A-saccharin cocrystal Form 1. In a subembodiment, Compound A-saccharin cocrystal Form 1 has an XRPD pattern substantially as shown in Figure 86. In yet another subembodiment, Compound A-saccharin cocrystal Form 1 has a DSC pattern substantially as shown in Figure 87. In yet another subembodiment, Compound A-saccharin cocrystal Form 1 has a DVS pattern substantially as shown in Figure 88.

[0088] In embodiment 28, the invention provides an L-tartaric acid cocrystal (compound AL-tartaric acid). In embodiment 28a, the invention provides a solid form of compound AL-tartaric acid. In embodiment 28b, the solid form of compound AL-tartaric acid is crystalline compound AL-tartaric acid cocrystal Form 1. In a subembodiment, compound AL-tartaric acid cocrystal Form 1 has an XRPD pattern substantially as shown in Figure 89. In yet another subembodiment, compound AL-tartaric acid cocrystal Form 1 has a DSC pattern substantially as shown in Figure 90. In yet another subembodiment, compound AL-tartaric acid cocrystal Form 1 has a DVS pattern substantially as shown in Figure 91.

[0089] In embodiment 29, the invention provides a urea co-crystal (Compound A-urea). In embodiment 29a, the invention provides a solid form of Compound A-urea. In embodiment 29b, the solid form of Compound A-urea is crystalline Compound A-urea co-crystal Form 1. In a subembodiment, Compound A-urea co-crystal Form 1 has an XRPD pattern substantially as shown in Figure 92. In yet another subembodiment, Compound A-urea co-crystal Form 1 has a DSC pattern substantially as shown in Figure 93. In yet another subembodiment, Compound A-urea co-crystal Form 1 has a DVS pattern substantially as shown in Figure 94.

[0090] In embodiment 30, the present invention provides a pharmaceutical composition comprising a salt, hydrate, solvate, or co-crystal of Compound A; or a solid form of Compound A, its salt, hydrate, solvate, or co-crystal.

[0091] In embodiment 30a, the present invention provides a pharmaceutical composition comprising a solid form of Compound A, a salt, hydrate, solvate, or co-crystal of Compound A. In a subembodiment, the solid form is crystalline or amorphous. In a subembodiment, the solid form is crystalline Compound A-Form 1. In another subembodiment, the solid form is a crystalline form of anhydrous Compound A, including crystalline anhydrous Forms 3, 4, 5, 6, 7, or 8.

[0092] In embodiment 30b, the present invention provides a compound A hydrochloride salt (Compound A-HCl), a mesylate salt (Compound A-MsA), a tosylate salt (Compound A-TsA), a sulfate salt (Compound A-Sulfate), a variable hydrate salt (Compound A-Variable Hydrate), a tetrahydrofuran solvate (Compound A-THF), an ethanol solvate (Compound A-Ethanol), a 1-propanol solvate (Compound A-1-Propanol), an isopropyl alcohol solvate (Compound A-IPA), a methanol solvate (Compound A-Methanol), an isopropyl acetate solvate (Compound A-IPAc), an acetone solvate (Compound A-Acetone), a cyclopentyl methyl ether solvate (Compound A-CPME), a dioxane solvate (Compound A-Dioxane), an ethyl acetate solvate (Compound A-EtOAc), an acetonitrile solvate (Compound A-Ethyl Acetate ... Provided is a pharmaceutical composition comprising a salt, hydrate, solvate, or co-crystal of Compound A selected from a solvate (Compound A-MeCN), a methyl tert-butyl ether solvate (Compound A-MTBE) a toluene solvate (Compound A-toluene), a dodecyl sulfate salt (Compound A-dodecyl sulfate), a dimethylformamide (DMF) solvate hydrate (Compound A-DMF-hydrate), a dimethylacetamide (DMAC) solvate (Compound A-DMAC), a monobesylate salt hydrate (Compound A-besylate-hydrate), a caffeine co-crystal (Compound A-caffeine), a citric acid co-crystal (Compound A-citric acid), a saccharin co-crystal (Compound A-saccharin), an L-tartaric acid co-crystal (Compound A-AL-tartaric acid), or a urea co-crystal (Compound A-urea); or a solid form thereof.

[0093] In embodiment 30c, the present invention provides a pharmaceutical composition comprising a solid form of Compound A-HCl of any of embodiments 1a-1j or any of its subembodiments, and a pharma- ceutically acceptable excipient. Preferably, the solid form of Compound A-HCl is crystalline Form 1 of Compound A-HCl having an XRPD pattern substantially as shown in Figure 1.

[0094] In embodiment 30d, the present invention provides a pharmaceutical composition comprising a solid form of compound A-MsA of any of embodiments 2a-2j or any of its subembodiments, and a pharma- ceutically acceptable excipient. Preferably, the solid form of compound A-MsA is crystalline Form 1 of compound A-MsA having an XRPD pattern substantially as shown in Figure 10.

[0095] In embodiment 30e, the present invention provides a pharmaceutical composition comprising a solid form of compound A-TsA of any of embodiments 3a-3j or any of its subembodiments, and a pharma- ceutically acceptable excipient. Preferably, the solid form of compound A-TsA is crystalline Form 4 of compound A-TsA having an XRPD pattern substantially as shown in Figure 20.

[0096] In embodiment 30f, the present invention provides a pharmaceutical composition comprising a solid form of Compound A-variable hydrate of any of embodiments 6a-6e or any of its sub-embodiments, and a pharma- ceutically acceptable excipient. Preferably, the Compound A-variable hydrate is Compound A-variable hydrate Form 2 having an XRPD pattern substantially as shown in Figure 36.

[0097] In embodiment 30g, the present invention provides a pharmaceutical composition comprising a crystalline anhydrous form of Compound A and a pharma- ceutically acceptable excipient. Preferably, the crystalline anhydrous form of Compound A has an XRPD pattern substantially as shown in any one of Figures 40, 43, 44, 47, 49, or 50.

[0098] In embodiment 30h, the present invention provides a pharmaceutical composition comprising a solid form of Compound A-citric acid co-crystal Form 1 and a pharma- ceutically acceptable excipient. Preferably, Compound A-citric acid co-crystal Form 1 has an XRPD pattern substantially as shown in Figure 82.

[0099] In embodiment 30i, the present invention provides a pharmaceutical composition comprising a solid form of Compound A-citric acid co-crystal Form 2 and a pharma- ceutically acceptable excipient. Preferably, Compound A-citric acid co-crystal Form 2 has an XRPD pattern substantially as shown in Figure 84.

[0100] In embodiment 30j, the present invention provides a pharmaceutical composition comprising a solid form of Compound A-dodecyl sulfate and a pharma- ceutically acceptable excipient. Preferably, Compound A-dodecyl sulfate has an XRPD pattern substantially as shown in Figure 71.

[0101] In embodiment 31, the present invention provides a method for treating a subject suffering from a disease mediated by KIF18A inhibition, comprising administering to a subject in need thereof a pharma- ceutical composition described in any one of embodiments 30 to 30j.

[0102] In embodiment 31a, the present invention provides the method according to embodiment 31, wherein the disease mediated by KIF18A inhibition is a neoplastic disease. In a subembodiment, the neoplastic disease is a cancer or tumor. In a further subembodiment, the cancer is ovarian cancer, breast cancer, lung cancer, or endometrial cancer. In a further subembodiment, the ovarian cancer is high-grade serous ovarian cancer (HGSOC), optionally metastatic or unresectable HGSOC. In a further subembodiment, the HGSOC is platinum-resistant HGSOC, or the HGSOC has progressed during or within 6 months of a platinum-containing regimen. In a further subembodiment, the cancer is primary peritoneal cancer and / or fallopian tube cancer. In a further subembodiment, the breast cancer is triple-negative breast cancer. In a further subembodiment, the endometrial cancer is serous endometrial cancer. In a further subembodiment, the serous endometrial cancer is metastatic or recurrent serous endometrial cancer. In a further subembodiment, the serous endometrial cancer has relapsed or is refractory to at least one line of systemic chemotherapy. In a further subembodiment, the serous endometrial cancer has relapsed or is refractory to at least one line of systemic chemotherapy. In a further subembodiment, the lung cancer is non-small cell lung cancer. In a further subembodiment, the tumor is an advanced solid tumor. In a further subembodiment, the tumor is unresectable, metastatic and / or non-localized. In a further subembodiment, the tumor has relapsed or is refractory to at least one line of systemic chemotherapy.

[0103] In embodiment 31b, the present invention provides a method according to embodiment 31, 31a, or any of their sub-embodiments, wherein the subject relapses or is refractory to at least one line of systemic chemotherapy. In a sub-embodiment, the systemic chemotherapy comprises taxane, gemcitabine, or doxorubicin. In a further sub-embodiment, the systemic chemotherapy comprises cisplatin, carboplatin, or levantinib.

[0104] In embodiment 31c, the present invention provides a method according to embodiment 31, 31a, 31b, or any sub-embodiment thereof, wherein the cancer or tumor comprises cells positive for an inactivated TP53 gene and / or positive for at least one of an inactivated Rb gene, (ii) an amplified CCNE1 gene or an overexpressed CCNE1 gene product, (iii) an inactivated BRCA gene, or (iv) a combination thereof.

[0105] In embodiment 31d, the invention provides the method of embodiment 31, 31a, 31b, 31c, or any sub-embodiment thereof, wherein the subject is an adult human.

[0106] In embodiment 32, the present invention provides a method of preparing Compound A-HCl of any of embodiments 1-1j, or any of its subembodiments, comprising combining hydrochloric acid, Compound A, and a suitable solvent to form Compound A-HCl salt, or a solid form thereof. In subembodiments, the suitable solvent is selected from acetonitrile / water, acetonitrile / 1,4-dioxane, tetrahydrofuran / water, N-methyl-2-pyrrolidone / ethanol, or acetone / water.

[0107] In embodiment 33, the present invention provides a method of preparing compound A-MsA of any of embodiments 2-2j, or any of their subembodiments, comprising combining methanesulfonic acid, compound A, and a suitable solvent to form a compound A-MsA salt or solid form thereof. In subembodiments, the suitable solvent is selected from acetonitrile or ethyl acetate.

[0108] In embodiment 34, the present invention provides a method for preparing Compound A-TsA of any of embodiments 3-3i, or any of its sub-embodiments, comprising combining p-toluenesulfonic acid, Compound A, and a suitable solvent to form Compound A-TsA salt or a solid form thereof. In a sub-embodiment, the suitable solvent is selected from ethanol or isopropanol.

[0109] In embodiment 35, the present invention provides a method for preparing a solid form of Compound A-variable-hydrate-form 2 of any of embodiments 6b-6h, or any of its subembodiments, comprising (a) combining water with a mixture of Compound A-methanol solvate form 1 and Compound A-ethanol solvate form 1 to form Compound A-variable-hydrate-form 2, or (b) combining Compound A in an alcoholic solvent, followed by removal of the alcoholic solvent by aqueous treatment, filtration, and drying at elevated temperature. In a subembodiment, the alcoholic solvent is a mixture of methanol and ethanol. In another subembodiment, the elevated temperature is 50°C.

[0110] In embodiment 36, the present invention provides a hydrochloride salt of Compound A, having the structure shown below: [ka]

[0111] In embodiment 37, the present invention provides a mesylate salt of Compound A, having the structure shown below: [ka]

[0112] In embodiment 38, the present invention provides a tosylate salt of Compound A, having the structure shown below: [ka]

[0113] In embodiment 39, the present invention provides a hydrate of Compound A having the structure: [ka] where n ranges from 0.5 to 2.

[0114] Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this disclosure belongs. Methods and materials are described herein for use in this disclosure, but other suitable methods and materials known in the art can also be used. The materials, methods, and examples are illustrative only and are not intended to be limiting. All publications, patent applications, patents, sequences, database entries, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control.

[0115] Further aspects and advantages will be apparent to those of ordinary skill in the art upon review of the following detailed description in conjunction with the drawings, in which: The following description includes specific embodiments, with the understanding that the disclosure is by way of example and is not intended to limit the invention to the specific embodiments described herein. [Brief description of the drawings]

[0116] [Figure 1] 1 shows the X-ray powder diffraction ("XRPD") pattern of crystalline Compound A-HCl Form 1. [Diagram 2] 1 shows a differential scanning calorimetry (DSC) thermograph and thermogravimetric analysis (TGA) of crystalline Compound A-HCl Form 1. [Diagram 3] FIG. 1 shows the dynamic vapor sorption (DVS) profile of crystalline Compound A-HCl Form 1. [Figure 4] 1 shows the solid state 19F NMR of crystalline Compound A-HCl Form 1. [Diagram 5] 1 shows the single crystal X-ray crystal structure of crystalline Compound A-HCl Form 1. [Figure 6]1 shows powder dissolution of Compound A-anhydrous Form 3, Compound A-variably-hydrated Form 2, Compound A-HCl Form 1, and Compound A-amorphous Form. [Figure 7] 1 shows the XRPD pattern of crystalline Compound A-HCl Form 2. [Figure 8] 1 shows a DSC thermograph of crystalline Compound A-HCl Form 2. [Figure 9] 1 shows modulated DSC of the amorphous form of Compound A-HCl. [Figure 10] 1 shows the XRPD pattern of crystalline compound A-MsA Form 1. [Figure 11] 1 shows the DSC thermograph and TGA of crystalline compound A-MsA Form 1. [Figure 12] 1 shows the DVS moisture sorption profile of crystalline Compound A-MsA Form 1. [Figure 13] 1 shows the solid-state 19F NMR of crystalline compound A-MsA Form 1. [Figure 14] 1 shows the XRPD pattern of crystalline compound A-MsA Form 2. [Figure 15] 1 shows a DSC thermogram of crystalline compound A-MsA Form 2. [Figure 16] 1 shows the TGA of crystalline compound A-MsA Form 2. [Figure 17] 1 shows the XRPD pattern of crystalline compound A-TsA Form 1. [Figure 18] 1 shows variable temperature X-ray diffraction (VTXRD) of crystalline compound A-TsA Form 1, which recrystallizes above 180° C. to form new crystalline compound A-TsA Form 5. [Figure 19] 1 shows the DSC thermograph and TGA of crystalline compound A-TsA Form 1. [Figure 20] 1 shows the XRPD pattern of crystalline compound A-TsA Form 2. [Figure 21] 1 shows the DSC thermograph and TGA of crystalline compound A-TsA Form 2. [Figure 22] 1 shows the XRPD pattern of crystalline compound A-TsA Form 3. [Diagram 23]1 shows the DSC thermograph and TGA of crystalline compound A-TsA Form 3. [Figure 24A] 1 shows the XRPD pattern of crystalline compound A-TsA Form 4. [Figure 24B] 1 shows the single crystal X-ray crystal structure of crystalline compound A-TsA Form 4. [Diagram 25] 1 shows the DSC thermograph and TGA of crystalline compound A-TsA Form 4. [Figure 26] 1 shows the solid-state 19F NMR of crystalline compound A-TsA Form 4. [Figure 27] 1 shows the XRPD pattern of crystalline compound A-TsA Form 5. [Figure 28] 1 shows the XRPD pattern of crystalline compound A-DiTsA form 6. [Figure 29] 1 shows powder dissolution and kinetic solubility of Compound A-HCl salt Form 1, Compound A-mesylate salt Form 1, and Compound A-tosylate salt Form 4 in fasted simulated small intestinal fluid (FaSSIF). [Diagram 30] 1 shows the XRPD pattern of crystalline Compound A-Sulfate Salt Form 1. [Diagram 31] 1 shows a DSC thermograph and TGA of crystalline Compound A-Sulfate Salt Form 1. [Diagram 32] 1 shows the DVS of crystalline Compound A-Sulfate Salt Form 1. [Diagram 33] FIG. 1 shows the XRPD pattern of Compound A-amorphous form. [Diagram 34] FIG. 1 shows a DSC thermograph showing that Compound A-amorphous form has a glass transition temperature (Tg) of 91° C. [Diagram 35] 1 shows a TGA-IR showing approximately 1.05% water weight loss from the amorphous form of Compound A below 100° C. [Diagram 36] 1 shows the XRPD pattern of crystalline Compound A-variable hydrate form 2. [Figure 37] 1 shows a DSC thermogram of crystalline Compound A-variable hydrate form 2. [Figure 38] 1 shows the TGA of crystalline Compound A-variable hydrate form 2. [Figure 39] 1 shows the DVS moisture sorption profile of crystalline Compound A-variable hydrate form 2. [Diagram 40] 1 shows the XRPD pattern of crystalline Compound A-anhydrous Form 3. [Diagram 41] 1 shows a DSC thermogram of crystalline Compound A-anhydrous Form 3. [Diagram 42] 1 shows the DVS profile of crystalline Compound A-anhydrous Form 3. [Diagram 43] 1 shows the XRPD pattern of crystalline Compound A-anhydrous Form 4. [Diagram 44] 1 shows the XRPD pattern of crystalline Compound A-anhydrous Form 5. [Diagram 45] 1 shows the DSC thermograph and TGA of crystalline Compound A-anhydrous Form 5. [Figure 46] 1 shows the DVS profile of crystalline Compound A-anhydrous Form 5, where anhydrous Form 5 rehydrates to Compound A-monohydrate. [Figure 47] 1 shows the XRPD pattern of crystalline Compound A-anhydrous Form 6. [Figure 48] 1 shows the DSC thermograph and TGA of crystalline Compound A-anhydrous Form 6. [Figure 49] 1 shows the XRPD pattern of crystalline Compound A-anhydrous Form 7. [Figure 50] 1 shows the XRPD pattern of crystalline Compound A-anhydrous Form 8. [Figure 51] 1 shows the DSC thermograph and TGA of crystalline Compound A-anhydrous Form 8. [Figure 52] 1 shows the XRPD pattern of crystalline Compound A Form 1. [Figure 53] 1 shows the XRPD pattern of crystalline Compound A-THF solvate. [Figure 54] 1 shows a DSC thermograph and TGA of crystalline Compound A-THF solvate. [Figure 55] 1 shows the XRPD pattern of crystalline Compound A-ethanol solvate. [Figure 56] 1 shows a TGA of crystalline Compound A-ethanol solvate. [Figure 57]1 is a DSC thermograph of crystalline Compound A-ethanol solvate. [Figure 58] Shown is the XRPD pattern of crystalline compound A-1-propanol solvate. [Figure 59] 1 shows a DSC thermograph and TGA of crystalline Compound A-1-propanol solvate. [Figure 60] 1 shows the XRPD pattern of crystalline Compound A-isopropyl alcohol (IPA) solvate. [Figure 61] 1 shows a DSC thermograph and TGA of crystalline Compound A-IPA solvate. [Figure 62] 1 shows the XRPD pattern of crystalline Compound A-methanol solvate. [Figure 63] FIG. 1 shows the XRPD pattern of crystalline Compound A-isopropyl acetate (IPAc) solvate. [Figure 64] 1 shows the XRPD pattern of crystalline Compound A-acetone solvate. [Figure 65] 1 shows the XRPD pattern of crystalline Compound A-cyclopentyl methyl ether (CPME) solvate. [Figure 66] 1 shows the XRPD pattern of crystalline Compound A-dioxane solvate. [Figure 67] FIG. 1 shows the XRPD pattern of crystalline Compound A-ethyl acetate (EtOAc) solvate. [Figure 68] 1 shows the XRPD pattern of crystalline Compound A-acetonitrile (MeCN) solvate. [Figure 69] 1 shows the XRPD pattern of crystalline Compound A - methyl tert-butyl ether (MTBE) solvate. [Figure 70] 1 shows the XRPD pattern of crystalline Compound A-toluene solvate. [Figure 71] 1 shows the XRPD pattern of crystalline Compound A-dodecyl sulfate solvate. [Figure 72] 1 shows a DSC thermograph and TGA of crystalline Compound A-dodecyl sulfate solvate. [Figure 73]1 shows the XRPD pattern of crystalline Compound A-dimethylformamide (DMF) solvate hydrate. [Figure 74] 1 shows a DSC thermogram of crystalline Compound A-dimethylformamide (DMF) solvate hydrate. [Figure 75] 1 shows the XRPD pattern of crystalline Compound A-dimethylacetamide (DMAC) solvate. [Figure 76] 1 is a DSC thermograph of crystalline Compound A-dimethylacetamide (DMAC) solvate. [Figure 77] 1 shows the XRPD pattern of crystalline Compound A-monobesylate hydrate Form 1. [Figure 78] 1 shows a DSC thermograph and TGA of crystalline Compound A-monobesylate hydrate Form 1. [Figure 79] 1 shows the XRPD pattern of crystalline Compound A-caffeine cocrystal Form 1. [Figure 80] 1 shows the DSC thermograph and TGA of crystalline Compound A-caffeine cocrystal Form 1. [Figure 81] 1 shows the DVS profile of crystalline Compound A-caffeine cocrystal Form 1. [Figure 82] FIG. 1 shows the XRPD pattern of crystalline Compound A-citric acid co-crystal Form 1. [Figure 83] 1 shows the DSC thermograph and TGA of crystalline Compound A-citric acid co-crystal Form 1. [Figure 84] FIG. 2 shows the XRPD pattern of crystalline Compound A-citric acid co-crystal Form 2. [Figure 85] 1 shows the DSC thermograph and TGA of crystalline Compound A-citric acid co-crystal Form 2. [Figure 86] 1 shows the XRPD pattern of crystalline Compound A-saccharin cocrystal Form 1. [Figure 87] 1 shows the DSC thermograph and TGA of crystalline Compound A-saccharin cocrystal Form 1. [Figure 88] 1 shows the DVS profile of crystalline Compound A-saccharin cocrystal Form 1. [Figure 89]FIG. 1 shows the XRPD pattern of crystalline compound AL-tartaric acid co-crystal Form 1. [Figure 90] 1 shows the DSC thermograph and TGA of the crystalline compound AL-tartaric acid co-crystal Form 1. [Figure 91] FIG. 1 shows the DVS profile of the crystalline compound AL-tartaric acid co-crystal Form 1. [Figure 92] FIG. 1 shows the XRPD pattern of crystalline Compound A-urea co-crystal Form 1. [Figure 93] 1 shows the DSC thermograph and TGA of crystalline Compound A-urea co-crystal Form 1. [Figure 94] FIG. 1 shows the DVS profile of crystalline Compound A-urea cocrystal Form 1. [Figure 95] FIG. 1 shows a crossover PK study in dogs of Compound A-HCl Form 1, Compound A-anhydrous Form 3, and Compound A-amorphous. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0117] Provided herein are salts, hydrates, solvates or cocrystals of Compound A; solid forms of Compound A, salts, hydrates, solvates or cocrystals thereof; pharmaceutical compositions thereof; and a method of treating a subject suffering from cancer, comprising administering a therapeutically effective amount of the pharmaceutical composition to the subject.

[0118] Compound A is a KIF18A inhibitor, and in various embodiments has a KIF18A ATPase IC50 of about 0.071 μM. The KIF18A gene belongs to the kinesin-8 subfamily and is a plus-end directed motor. KIF18A is thought to affect the dynamics of the plus-ends of kinetochore microtubules, controlling the correct positioning of chromosomes and spindle tension. Deletion of human KIF18A results in longer spindles, increased chromosome oscillations in metaphase, and activation of the mitotic spindle assembly checkpoint in HeLa cervical cancer cells (MI Mayr et al, Current Biology 17, 488-98, 2007). KIF18A is overexpressed in various cancers, including but not limited to colon cancer, breast cancer, lung cancer, pancreatic cancer, prostate cancer, bladder cancer, head cancer, neck cancer, cervical cancer, and ovarian cancer. Overexpression of KIF18A attenuates sister chromatid fluctuations, resulting in a rigid metaphase plate. gcd2 / gcd2Inactivation of KIF18A motor function by mutagenic ethyl methanesulfonate (EMS) treatment in mice (missense mutation in the motor domain (R308K)) results in viable mice with no gross abnormalities in major organs except for obvious testicular atrophy and sterility (J Stumpff et al Developmental Cell. 2008;14:252-262; J Stumpff et al Developmental Cell. 2012;22:1017-1029; XS Liu et al. Genes & Cancer. 2010;1:26-39; CL Fonseca et al J Cell Biol. 2019;1-16; A Czechanski et al Developmental Biology. 2015;402:253-262. O Rath,F Kozielski.Nature Reviews Cancer. 2012;12:527-539). Normal human and mouse KIF18A-deficient somatic cells complete cell division with relatively normal mitotic progression but imprecise chromosome alignment, resulting in daughter cells with normal karyotypes, and some defects in exit from mitosis are observed in a subset of normal cells, resulting in micronucleation during slower proliferation (CL Fonseca et al J Cell Biol.2019;1-16). These genetic studies suggest that normal germ and somatic cells have different dependencies on the requirements for chromosome alignment, and indicate that KIF18A may be dispensable for normal euploid somatic cell division (XS Liu et al Genes & Cancer.2010;1:26-39;A Czechanski et al Developmental Biology.2015;402:253-262). In normal human tissues, KIF18A expression is elevated in tissues with actively cycling cells, with highest expression in testis (GTEx Portal, GTEx Portal, J Lonsdale et al Nature Genetics. 2013:29; 45:580).In various embodiments, compound A inhibits ATPase activity.For example, compound A inhibits MT-ATPase activity but not basal ATPase activity.

[0119] The compounds disclosed herein are identified herein by their chemical structure and / or chemical name. In the event that the chemical structure and the chemical name conflict, the chemical structure is determinative of the compound's identity.

[0120] When ranges are used herein for physical properties, such as molecular weight, or chemical properties, such as chemical formula, all combinations and subcombinations of the ranges and specific embodiments are intended to be included.

[0121] As used herein, dashed and bold bonds (i.e. [ka] and [ka] Chemical structures containing one or more stereocenters depicted with a solid line are meant to depict the absolute stereochemistry of the stereocenter(s) present in the chemical structure. As used herein, bonds symbolized with a simple line do not depict stereo preference. Unless otherwise stated to the contrary, chemical structures containing one or more stereocenters illustrated herein without depicting absolute or relative stereochemistry encompass all possible stereoisomers (e.g., diastereomers, enantiomers) of the compound and mixtures thereof. Structures with a single bold or dashed line and at least one additional simple line encompass a single enantiomeric series of all possible diastereomers.

[0122] The term "about" is intended to account for variations due to experimental error. All measurements reported herein are understood to be modified by the term "about," unless expressly stated otherwise, whether or not the term is explicitly used. As used herein, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise.

[0123] The term "compound," as used herein, is intended to include all stereoisomers, geometric isomers, tautomers, and isotopes of the structures depicted. A compound identified herein by name or structure as one particular tautomer is intended to include the other tautomers, unless otherwise specified.

[0124] The term "hydrate" refers to a chemical entity formed by the interaction of water with a compound, including, for example, hemihydrate, monohydrate, dihydrate, trihydrate, etc. Hydrates, as used herein, can have variable amounts of water, typically 0.5 to 2 water molecules, e.g., 0.5, 1, 1.5, or 2 water molecules per Compound A molecule, and are referred to as "variable hydrates." The number of water molecules can vary with different preparation methods and storage conditions of the hydrate form.

[0125] The terms "solid form" and "physical form" are intended to include all crystalline and amorphous forms of a compound, including, for example, polymorphs, pseudopolymorphs, solvates, hydrates, nonsolvated polymorphs (including anhydrates), conformational polymorphs, and amorphous forms, as well as mixtures thereof, unless a specific crystalline or amorphous form is referenced.

[0126] The term "cocrystal," as used herein, refers to a crystalline complex in which a neutral molecular component and Compound A are bound in a crystal lattice by non-covalent interactions, often including hydrogen bonds. Examples of cocrystals include caffeine cocrystal (Compound A-caffeine), citric acid cocrystal (Compound A-citric acid), saccharin cocrystal (Compound A-saccharin), L-tartaric acid cocrystal (Compound A-tartaric acid), or urea cocrystal (Compound A-urea).

[0127] "Glass transition temperature" refers to the temperature range over which an amorphous solid form gradually and reversibly transitions from a hard, relatively brittle "glassy" state to a viscous or rubbery state with increasing temperature.

[0128] Salts, hydrates, solvates, co-crystals of Compound A; and isolation and purification of solid forms of Compound A, its salts, solvates, and co-crystals, including crystalline anhydrous forms. Compound A has an ionizable functional group with a weakly basic pKa value of 3.9 and a weakly acidic pKa value of 7.3. From a high-throughput manual polymorph screening, we have generated various Compound A salts, hydrates, solvates, co-crystals; as well as solid forms of Compound A, its salts, hydrates, solvates, and co-crystals, including a crystalline anhydrous form. Desolvation of the ethanol solvate by drying produced the relatively stable Compound A-Hydrate-Form 2, which began to dehydrate at 25°C and had very low solubility. Desolvation of the Compound A-THF solvate produced the anhydrous Compound A-Form 3, which quickly transformed into Compound A Hydrate-Form 2 in aqueous media or upon uptake of water. Based on the solid-state properties of the free base form of Compound A and their propensity to form solvates, the inventors have generated a variety of salts, hydrates, solvates and co-crystals of Compound A that may be suitable for scale-up and crystallization of the drug substance for pharmaceutical development.

[0129] From the bulk manual salt screen, various counterions and solvents were tested to produce crystalline salts and solvates of Compound A. Various salts, hydrates, and solvates of Compound A, including sulfate, besylate, mesylate, and tosylate, were formed in multiple polymorphic forms. Further solubility and stability studies led to Compound A-HCl salt Form 1 being prioritized for further evaluation due to its acceptable solubility and stability profile, improved biopharmaceutical properties, and favorable crystallization process. Polymorph screening of Compound A-HCl salt Form 1 yielded a total of 126 crystalline samples from 384 crystallization conditions. Of these, the XRPD patterns of 90 samples were identical to that of HCl salt Form 1. The others may contain disproportionated forms of Compound A or solvates of Compound A. Polymorph screening of Compound A-HCl salt revealed that Compound A-HCl-Form 1 was the most thermodynamically stable form. Several co-crystals of Compound A were also generated from the co-crystal screen, for example, co-crystals with citric acid, tartaric acid, caffeine and urea.

[0130] Crystalline Compound A-Hydrochloride Salt Form 1 (Compound A-HCl Salt Form 1) Also provided herein is crystalline Compound A-HCl salt Form 1. Crystalline Compound A-HCl salt Form 1 is a solid-state salt obtained as described in the Examples. 19 In some embodiments, the crystalline Compound A-HCl salt Form 1 can be characterized by a solid-state NMR peak at -91 and -103 ± 0.5 ppm. 19 F NMR, where "substantially" means that the reported peaks may vary by ±0.5 ppm.

[0131] Crystalline Compound A-HCl salt Form 1 may be further characterized by a powder X-ray diffraction pattern using CuKα radiation obtained as described in the Examples, having peaks at 7.5, 16.9, and 20.2±0.2 degrees 2θ. Crystalline Compound A-HCl Form 1 may optionally be further characterized by a powder X-ray diffraction pattern using CuKα radiation, having additional peaks at 12.8, 18.2, 22.7, 23.6, 24.8, and 26.1±0.2 degrees 2θ. Crystalline Compound A-HCl Form 1 may optionally be further characterized by a powder X-ray diffraction pattern using CuKα radiation, having additional peaks at 10.9, 14.5, 15.7, 15.9, 19.8, 20.6, 21.6, 23.2, 26.1, and 26.8±0.2 degrees 2θ. In some embodiments, crystalline Compound A-HCl salt Form 1 has a powder X-ray diffraction pattern substantially as shown in Figure 1, where "substantially" means that the reported peaks may vary by ±0.2°. Those skilled in the art of XRPD know that the relative peak heights in a spectrum depend on several factors such as sample preparation and instrument geometry, but that the positions of the peaks are relatively independent of experimental details.

[0132] A differential scanning calorimetry (DSC) thermogram was obtained for crystalline Compound A-HCl salt Form 1 as described in the Examples. The DSC curve exhibited an endothermic transition at 271.5° C.±3° C. Thus, in some embodiments, crystalline Compound A-HCl salt Form 1 may be characterized by a DSC thermogram having an endothermic transition with an onset between 268.5° C. and 274.5° C. For example, in some embodiments, crystalline Compound A-HCl salt Form 1 is characterized by a DSC as shown in FIG. 2.

[0133] Crystalline Compound A-HCl salt Form 1 may also be characterized by thermogravimetric analysis (TGA). Thus, crystalline Compound A-HCl salt Form 1 may be characterized by an approximately 4% weight loss with an onset temperature of 268.3° C. to 273.7° C. For example, crystalline Compound A-HCl salt Form 1 may be characterized by an approximately 4% weight loss up to approximately 271° C. In some embodiments, crystalline Compound A-HCl salt Form 1 has a thermogravimetric analysis substantially as shown in FIG. 2, where "substantially" means that the reported TGA characteristics may vary by ±1% of the approximately 4% weight loss.

[0134] Crystalline Compound A-HCl salt Form 1 can be characterized by a moisture sorption profile. For example, in some embodiments, crystalline Compound A-HCl salt Form 1 is characterized by a moisture sorption profile (DVS) as shown in Figure 3, exhibiting less than 0.5% weight gain at 95% RH.

[0135] Crystalline Compound A-HCl salt Form 1 is further characterized by a single crystalline structure substantially as shown in FIG. 5, or in the Examples.

[0136] Further provided herein is a pharmaceutical composition comprising the crystalline Compound A-HCl salt Form 1 described herein and a pharma- ceutically acceptable excipient.

[0137] Crystalline Compound A-Mesylate Form 1 (Compound A-MsA Salt Form 1) Also provided herein is crystalline Compound A-MsA salt Form 1. Crystalline Compound A-MsA salt Form 1 is a solid-state salt obtained as described in the Examples. 19 In some embodiments, the crystalline Compound A-MsA Salt Form 1 can be characterized by a solid-state peak at -95.2 and -103.2±0.5 ppm as shown in FIG. 19 F NMR, where "substantially" means that the reported peaks may vary by ±0.5 ppm.

[0138] Crystalline compound A-MsA salt Form 1 may be further characterized by a powder X-ray diffraction pattern using CuKα radiation obtained as described in the Examples, having peaks at 7.0, 16.5, and 23.9±0.2°2θ. Crystalline compound A-MsA salt Form 1 may optionally be further characterized by a powder X-ray diffraction pattern using CuKα radiation, having additional peaks at 12.6, 15.7, 17.4, 18.5, 20.0, and 21.0±0.2°2θ. Crystalline compound A-MsA salt Form 1 may optionally be further characterized by a powder X-ray diffraction pattern using CuKα radiation, having additional peaks at 5.8, 11.8, 13.5, 15.3, 16.1, 18.0, 20.6, 25.2, 28.0, and 30.5±0.2°2θ. In some embodiments, crystalline Compound A-MsA Salt Form 1 has a powder X-ray diffraction pattern substantially as shown in Figure 10, where "substantially" means that the reported peaks may vary by ±0.2°. Those skilled in the art of XRPD know that the relative peak heights in a spectrum depend on several factors such as sample preparation and instrument geometry, but that the positions of the peaks are relatively independent of experimental details.

[0139] A differential scanning calorimetry (DSC) thermogram was obtained for crystalline compound A-MsA salt Form 1 as described in the Examples. The DSC curve exhibits an endothermic transition at 250° C.±3° C. Thus, in some embodiments, crystalline compound A-MsA salt Form 1 may be characterized by a DSC thermogram having an endothermic transition with an onset between 247° C. and 253° C. For example, in some embodiments, crystalline compound A-MsA salt Form 1 is characterized by a DSC as shown in FIG. 11.

[0140] Crystalline Compound A-MsA salt Form 1 may also be characterized by thermogravimetric analysis (TGA). Thus, crystalline Compound A-MsA salt Form 1 may be characterized by a weight loss of about 0.2% at an onset temperature of 247° C. to 253° C. For example, crystalline Compound A-MsA salt Form 1 may be characterized by a weight loss of about 0.2% up to about 250° C. In some embodiments, crystalline Compound A-MsA salt Form 1 has a thermogravimetric analysis substantially as shown in FIG. 11, where "substantially" means that the reported TGA characteristics may vary by ±1% of the 0.2% weight loss.

[0141] Crystalline Compound A-MsA salt Form 1 can be characterized by a moisture sorption profile. For example, in some embodiments, crystalline Compound A-MsA salt Form 1 is characterized by a moisture sorption profile as shown in Figure 12, exhibiting a weight gain of less than 1.2% at 95% RH.

[0142] Further provided herein is a pharmaceutical composition comprising the crystalline Compound A-MsA salt Form 1 described herein and a pharma- ceutically acceptable excipient.

[0143] Crystalline Compound A-Tosylate Form 1 (Compound-A-TsA Form 4) Also provided herein is crystalline Compound A-TsA salt Form 4. Crystalline Compound A-TsA salt Form 4 may be further characterized by a powder X-ray diffraction pattern using CuKα radiation obtained as described in the Examples, having peaks at 6.2, 14.7, and 23.5±0.2 degrees 2θ. Crystalline Compound A-TsA salt Form 4 may optionally be further characterized by a powder X-ray diffraction pattern using CuKα radiation, having additional peaks at 10.5, 12.4, 14.2, 19.1, 21.5, and 29.0±0.2 degrees 2θ. The crystalline compound A-TsA salt form 4 may optionally be further characterized by a powder X-ray diffraction pattern using CuKα radiation having additional peaks at 15.5, 16.5, 17.7, 18.3, 18.6, 20.1, 20.8, 24.1, and 25.3±0.2°2θ. In some embodiments, the crystalline compound A-TsA salt form 4 has a powder X-ray diffraction pattern substantially as shown in FIG. 24a, where "substantially" means that the reported peaks may vary by ±0.2°. Those skilled in the art of XRPD know that the relative peak heights of a spectrum depend on several factors, such as sample preparation and instrument geometry, but the positions of the peaks are relatively independent of experimental details.

[0144] Crystalline Compound A-TsA Salt Form 4 is further characterized by a single crystal structure substantially as shown in Figure 24b or as defined in the Examples.

[0145] A differential scanning calorimetry (DSC) thermogram was obtained for crystalline Compound A-TsA salt Form 4 as described in the Examples. The DSC curve exhibited an endothermic transition at 253° C.±3° C. Thus, in some embodiments, crystalline Compound A-TsA salt Form 4 may be characterized by a DSC thermogram having an endothermic transition with an onset between 250° C. and 256° C. For example, in some embodiments, crystalline Compound A-TsA salt Form 4 is characterized by DSC as shown in FIG. 25.

[0146] The crystalline compound A-TsA salt form 4 may also be characterized by thermogravimetric analysis (TGA). Thus, the crystalline compound A-TsA salt form 4 may be characterized by a weight loss of about 0.07% at an onset temperature of 250° C. to 256° C. For example, the crystalline compound A-TsA salt form 4 may be characterized by a weight loss of about 0.07% up to about 253° C. In some embodiments, the crystalline compound A-TsA salt form 4 has a thermogravimetric analysis substantially as depicted in FIG. 25, where "substantially" means that the reported TGA characteristics may vary by ±1% of the 0.07% weight loss.

[0147] Crystalline Compound A-TsA Salt Form 4 also has a solid-state 19 F NMR has peaks at -96.93 and -101.60 ppm, where "substantially" means that the reported peaks may vary by ±0.5 ppm.

[0148] Further provided herein is a pharmaceutical composition comprising the crystalline Compound A-HCl salt Form 4 described herein and a pharma- ceutically acceptable excipient.

[0149] Pharmaceutical Compositions Compound A as described in any of the above embodiments and subembodiments thereof can be combined with a pharma- ceutically acceptable excipient to provide a pharmaceutical formulation (also interchangeably referred to as a composition). The excipient can be a diluent or carrier. The formulation can affect the physical state, stability, in vivo release rate, and in vivo clearance rate of the administered drug. The phrases "pharmacologically acceptable" or "pharmacologically acceptable" refer to molecular entities and compositions that do not produce adverse, allergic, or other undesired reactions when administered to animals or humans. As used herein, "pharmacologically acceptable" includes any and all solvents, dispersion media, coating agents, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like. The use of such excipients for pharma- ceutical active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the therapeutic composition, its use in the therapeutic composition is contemplated. Supplementary active ingredients can also be incorporated into the composition. In an exemplary embodiment, the formulation comprises corn syrup solids, high oleic safflower oil, coconut oil, soybean oil, L-leucine, tribasic calcium phosphate, L-tyrosine, L-proline, L-lysine acetate, DATEM (emulsifier), L-glutamine, L-valine, dibasic potassium phosphate, L-isoleucine, L-arginine, L-alanine, glycine, L-asparagine monohydrate, L-serine, potassium citrate, L-threonine, sodium citrate, magnesium chloride, L-histidine, L-methionine, ascorbic acid, calcium carbonate, L-glutamic acid, L-cystine dihydrochloride, May contain L-tryptophan, L-aspartic acid, choline chloride, taurine, m-inositol, ferrous sulfate, ascorbyl palmitate, zinc sulfate, L-carnitine, alpha-tocopheryl acetate, sodium chloride, niacinamide, tocopherol mixture, calcium pantothenate, copper(II) sulfate, thiamine chloride hydrochloride, vitamin A palmitate, manganese sulfate, riboflavin, pyridoxine hydrochloride, folic acid, beta-carotene, potassium iodide, phylloquinone, biotin, sodium selenate, chromium chloride, sodium molybdate, vitamin D3, and cyanocobalamin.

[0150] For oral administration, Compound A may be combined with pharma- ceutically acceptable excipients, such as carriers well known in the art, to formulate a suitable composition. Such excipients and carriers allow Compound A to be formulated as tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions, etc., for oral ingestion by the patient to be treated. Oral pharmaceutical formulations can be obtained by adding solid excipients to Compound A, optionally grinding the resulting mixture, optionally adding suitable auxiliary agents, and then processing the mixture of granules to obtain tablet or dragee cores. Suitable excipients include, for example, fillers and cellulose preparations. Optionally, disintegrants can also be added. Pharmaceutically acceptable ingredients are well known for various types of formulations and can be, for example, binders (e.g., natural or synthetic polymers), lubricants, surfactants, sweeteners and flavorings, coating materials, preservatives, dyes, thickeners, adjuvants, antibacterial agents, antioxidants, and carriers for various formulation types.

[0151] When a therapeutically effective amount of Compound A is administered orally, the composition will typically be in the form of a solid (e.g., tablet, capsule, pill, powder, lozenge) or liquid (e.g., aqueous suspension, solution, elixir, syrup).

[0152] When administered in tablet form, the composition can further comprise a functional solid and / or solid carrier such as gelatin or an adjuvant.

[0153] When administered in liquid or suspension form, functional liquid and / or liquid carrier such as water, petroleum, oil of animal or plant origin can be added.The liquid form of the composition can further comprise physiological saline, sugar alcohol solution, dextrose or other sugar solution, or glycol.In one possible embodiment, the liquid carrier is non-aqueous or substantially non-aqueous.For administration in liquid form, the composition can be provided as a fast-dissolving solid formulation for dissolving or suspending immediately before administration.

[0154] For administration by inhalation, compound A can be delivered in the form of an aerosol spray from a pressurized pack or nebulizer using a suitable propellant. In pressurized aerosol embodiments, the dosage unit can be determined by providing a valve to deliver a metered amount. Capsules and cartridges, for example of gelatin, for use in an inhaler or insufflator can be formulated to contain a powder mix of the compound and a suitable powder base, such as lactose or starch.

[0155] In particular, Compound A can be administered orally, bucally, or sublingually in the form of tablets containing excipients such as starch or lactose, or in capsules or vaginally, alone or in combination with excipients, or in the form of elixirs or suspensions containing flavoring or coloring agents. Such liquid formulations can be prepared using pharma- ceutically acceptable additives, such as suspending agents.

[0156] safety In some embodiments, the method includes administering Compound A or a pharma- ceutically acceptable salt thereof in an amount that does not cause dose-limiting toxicity (DLT) during treatment with Compound A or a salt thereof. Optionally, the subject does not exhibit DLT associated with Compound A treatment during the treatment period. In various embodiments, the subject does not exhibit any grade 3 or grade 4 adverse events associated with Compound A treatment during the treatment period. In various embodiments, the treatment period is at least 2 weeks or at least 1 month (if not longer), for example, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, 1 year, 1.5 years, 2 years.

[0157] In representative embodiments, the method further comprises monitoring the subject's complete blood count before, during or after Compound A treatment. In various embodiments, the complete blood count comprises counting one or more of red blood cells, white blood cells, platelets, and neutrophils. Optionally, the complete blood count comprises measuring hematocrit and / or hemoglobin. In representative embodiments, the monitoring is performed weekly for about two months. In various embodiments, the subject's platelet count is greater than about 100,000 per μL of blood during Compound A treatment.

[0158] In representative embodiments, the method of the present disclosure is advantageously highly specific to cells of neoplastic disease. In various aspects, compound A effectively treats neoplastic disease, induces or enhances tumor regression, reduces tumor or cancer growth, or induces or enhances tumor or cancer cell death, without significant toxicity to normal somatic cells in the subject. In various aspects, compound A or a pharmaceutically acceptable salt thereof is administered in an amount effective to treat neoplastic disease, induces or enhances tumor regression, reduces tumor or cancer growth, and / or induces or enhances tumor or cancer cell death, without substantially reducing normal somatic cell growth in the subject. In representative embodiments, compound A or a pharmaceutically acceptable salt thereof is administered in an amount effective to treat neoplastic disease, induces or enhances tumor regression, reduces tumor or cancer growth, or induces or enhances tumor or cancer cell death, without substantially increasing apoptosis of normal somatic cells. As used herein, the term "normal" with respect to a cell refers to a cell that is not neoplastic and / or diseased. In various embodiments, the normal somatic cell is a human bone marrow mononuclear cell or a T cell. In various embodiments, the normal somatic cell is a TP53 MUT Not genetically characterized as TP53 WTIn various embodiments, compound A or a pharmaceutically acceptable salt thereof causes an increase in normal somatic cell apoptosis of 25% or less. In various embodiments, compound A or a pharmaceutically acceptable salt thereof causes a decrease in normal somatic cell proliferation of 25% or less in a subject. Optionally, the increase in normal somatic cell apoptosis or the decrease in normal somatic cell proliferation is less than about 20%, less than about 15%, less than about 10%, less than about 9%, less than about 8%, less than about 7%, less than about 6%, less than about 5%, less than about 4%, less than about 3%, less than about 2%, or less than about 1%.

[0159] The primary side effect of taxanes is myelosuppression, primarily neutropenia, with other side effects including peripheral edema, and neurotoxicity (peripheral neuropathy). In representative embodiments, the methods of the present disclosure treat a subject's neoplastic disease without causing any of these side effects observed in patients treated with taxanes, or treat a neoplastic disease in which such side effects are reduced in severity compared to those observed in patients treated with taxanes.

[0160] Treatment Efficacy As used herein, the term "treat" and related terms do not necessarily imply 100% or complete treatment. Rather, there are various degrees of treatment that one skilled in the art will recognize as having potential benefit or therapeutic effect. In this regard, the disclosed method of treating a neoplastic disease may provide any amount or level of treatment. Furthermore, the treatment provided by the disclosed method may include treatment of one or more conditions or symptoms or signs of the neoplastic disease being treated. The treatment provided by the disclosed method may also include slowing the progression of the neoplastic disease. For example, the method may treat a neoplastic disease by enhancing T cell activity or immune response to the neoplastic disease, reducing tumor or cancer growth or tumor burden, reducing metastasis of tumor cells, increasing cell death of tumor or cancer cells, or increasing tumor regression, etc. In accordance with the above, provided herein is a method of reducing tumor growth, tumor volume, or tumor burden, or increasing tumor regression in a subject. In an exemplary embodiment, the method includes administering compound A or a pharmaceutically acceptable salt thereof to the subject. As used herein, the terms "treat", "treating" and "treatment" refer to treatments including, but not limited to, prophylactic treatments, and preventative treatments. Prophylactic treatments generally consist of either preventing the onset of a disorder altogether or delaying the onset of a pre-clinically evident stage of the disorder in an individual.

[0161] In various embodiments, the method is by delaying the onset or recurrence of neoplastic disease, or by delaying the onset or onset of metastasis.In various embodiments, the method is by increasing the survival of the subject.In representative embodiments, onset or recurrence or onset is delayed at least 1 day, 2 days, 4 days, 6 days, 8 days, 10 days, 15 days, 30 days, 2 months, 3 months, 4 months, 6 months, 1 year, 2 years, 3 years, 4 years or more.

[0162] In various embodiments, the treatment provided by the methods of the present disclosure provides a therapeutic response according to the Response Evaluation Criteria in Solid Tumors (RECIST) or other similar criteria. RECIST is a set of criteria for assessing the progression, stabilization, or responsiveness of tumors and / or cancer cells, jointly developed by the National Cancer Institute, the National Cancer Institute of Canada Clinical Trials Group, and the European Organization for Research and Treatment of Cancer. According to RECIST, a particular tumor is measured at the start of an evaluation (e.g., a clinical trial) to provide a baseline for comparison after treatment with an agent. Tumor response evaluations and evaluation criteria are published in Eisenhauer et.al., Eur J Cancer 45:228-247 (2009) and Litiere et.al., Journal of Clinical Oncology 37(13):1102-1110 (2019) DOI:10.1200 / JCO.18.01100. In various embodiments, the treatment provided by the methods of the present disclosure provides a therapeutic response according to a modified RECIST tumor response evaluation, as follows: [Table 1] [Table 2]

[0163] In various embodiments, the subject exhibits at least stable disease (SD) after treatment with Compound A or a pharma- ceutically acceptable salt thereof. In various embodiments, the subject exhibits at least a partial response (PR) after treatment with Compound A or a pharma- ceutically acceptable salt thereof. In various embodiments, the subject exhibits a reduction in cancer antigen 125 (CA125) levels of at least 10%, at least 15%, at least 25%, at least 30%, at least 40%, or at least 50% compared to baseline. In representative embodiments, the subject exhibits a reduction in tumor volume of at least 10%, at least 15%, at least 25%, at least 30%, at least 40%, or at least 50% after treatment with Compound A.

[0164] Neoplastic disease As used herein, the term "neoplastic disease" refers to any condition that results in the growth of a tumor. In representative embodiments, the tumor is a benign tumor. In representative embodiments, the tumor is a malignant tumor. In various embodiments, the neoplastic disease is a tumor or cancer. In various embodiments, the cancer is acute lymphocytic carcinoma, acute myeloid leukemia, alveolar rhabdomyosarcoma, osteosarcoma, brain cancer, breast cancer, cancer of the anus, anal canal or anorectum, eye cancer, cancer of the intrahepatic bile duct, cancer of the joints, cancer of the neck, gallbladder or pleura, cancer of the nose, nasal cavity or middle ear, cancer of the oral cavity, cancer of the vulva, chronic lymphocytic leukemia, chronic myeloid cancer, colon cancer, esophageal cancer, cervical cancer, gastrointestinal cancer, or rectal cancer. The cancer is leucinoid tumor, Hodgkin's lymphoma, hypopharyngeal cancer, renal cancer, laryngeal cancer, liver cancer, lung cancer, malignant mesothelioma, melanoma, multiple myeloma, nasopharyngeal cancer, non-Hodgkin's lymphoma, ovarian cancer, pancreatic cancer, peritoneum, amniotic membrane, mesenteric cancer, pharyngeal cancer, prostate cancer, rectal cancer, kidney cancer (e.g. renal cell carcinoma (RCC)), small intestine cancer, soft tissue cancer, stomach cancer, testicular cancer, thyroid cancer, ureteral cancer, or bladder cancer. In certain embodiments, the cancer is head and neck cancer, ovarian cancer, cervical cancer, bladder cancer, esophageal cancer, pancreatic cancer, gastrointestinal cancer, stomach cancer, breast cancer, endometrial cancer, colorectal cancer, hepatocellular carcinoma, glioblastoma, bladder cancer, lung cancer, e.g. non-small cell lung cancer (NSCLC), or bronchioloalveolar carcinoma. In certain embodiments, the tumor is non-small cell lung cancer (NSCLC), head and neck cancer, renal cancer, triple negative breast cancer, or gastric cancer.In a representative embodiment, the subject has a tumor (e.g., solid tumor, hematological malignancy, or lymphatic malignancy), and the pharmaceutical composition is administered to the subject in an amount effective to treat the tumor of the subject.In another representative embodiment, the tumor is non-small cell lung cancer (NSCLC), small cell lung cancer (SCLC), head and neck cancer, renal cancer, breast cancer, melanoma, ovarian cancer, liver cancer, pancreatic cancer, colon cancer, prostate cancer, gastric cancer, lymphoma, or leukemia, and the pharmaceutical composition is administered to the subject in an amount effective to treat the tumor in the subject.

[0165] As used herein, the terms "cancer" and "cancerous" refer to or describe a physiological condition in a mammal that is typically characterized by uncontrolled cell proliferation. Examples of cancer include, but are not limited to, carcinoma, lymphoma, sarcoma, blastoma, and leukemia. More specific examples of such cancer include squamous cell carcinoma, lung cancer, pancreatic cancer, cervical cancer, bladder cancer, hepatoma, breast cancer, colon cancer, and head and neck cancer, ovarian cancer, and endometrial cancer. As used herein, the term "cancer" is not limited to any particular form of disease, but the method of the present invention is believed to be particularly effective against cancers that involve uncontrolled levels of KIF18A or are found to be dependent on KIF18A for proper chromosome segregation and survival in a mammal.

[0166] In various embodiments, the cancer is metastatic, the tumor is unresectable, or a combination thereof. In various embodiments, the cancer is an aneuploid cancer with chromosomal instability. In various embodiments, the neoplastic disease (e.g., cancer or tumor) comprises cells that are positive for inactivated TP53 gene, and / or positive for at least one of (i) inactivated Rb1 gene, (ii) amplified CCNE1 gene, CCNE1 gene copy number increase, or CCNE1 gene product overexpression, (iii) inactivated BRCA gene, or (iv) a combination thereof. In various embodiments, the neoplastic disease is triple-negative breast cancer (TNBC), non-luminal breast cancer (e.g., basal-like mesenchymal), or high-grade serous ovarian cancer (HGSOC). In various embodiments, the neoplastic disease is resistant or insensitive (insensitive) to treatment with CDK4 / 6 inhibitors. In various embodiments, the neoplastic disease is resistant or insensitive (insensitive) to treatment with a CDK4 / 6 inhibitor and is Rb1-proficient (vs. Rb1-deficient). In various embodiments, the neoplastic disease is resistant to treatment with a KIF18A inhibitor. In various embodiments, the neoplastic disease is resistant to treatment with a KIF18A inhibitor and is Rb1-deficient (vs. Rb1-proficient).

[0167] In representative embodiments, the neoplastic disease is breast cancer, optionally luminal breast cancer or TNBC. In various embodiments, the breast cancer is (a) histologically or cytologically confirmed to be metastatic or locally recurrent estrogen receptor (ER) negative (e.g., <1% by immunohistochemistry [IHC]), (b) progesterone receptor (PR) negative (e.g., <1% IHC), and (c) human epidermal growth factor receptor 2 (Her2) negative (either fluorescent in situ hybridization [FISH] negative, 0 or 1+ by IHC, or IHC 2+ and FISH negative, according to ASCO / CAP definition). In representative embodiments, the neoplastic disease is relapsed and / or refractory to at least one line of systemic chemotherapy in the metastatic setting, or intolerant to existing therapy(s) known to provide clinical benefit for the neoplastic disease. In representative embodiments, the cancer has been treated with an immune checkpoint inhibitor. In various embodiments, the breast cancer is hormone receptor (HR) positive and / or HER2 negative. In various embodiments, the breast cancer is advanced and / or metastatic breast cancer. In various embodiments, the breast cancer is HR+ / HER2- advanced or metastatic breast cancer that has progressed after endocrine therapy. In some embodiments, the breast cancer is hormone receptor positive (HR+) / HER2- advanced or metastatic breast cancer that has previously been treated with endocrine therapy and chemotherapy after the cancer has metastasized / grown. In various embodiments, the cancer is HR+ / HER2- advanced or metastatic breast cancer that has not been treated with hormone therapy (Arimidex (chemical name: anastrozole), Aromasin (chemical name: exemestane) and Femara (chemical name: letrozole). In various embodiments, the breast cancer is HR+ / HER2- advanced or metastatic breast cancer that has grown after being treated with hormone therapy. In various embodiments, the breast cancer is HER2 positive breast cancer, including but not limited to those similar to the HER2 positive breast cancer cells in Table 2. Optionally, the breast cancer is HER2 positive, estrogen receptor (ER) negative breast cancer. In various aspects, the neoplastic disease is ovarian cancer, optionally high grade serous ovarian cancer (HGSOC).Optionally, the ovarian cancer is platinum-resistant HGSOC. In representative embodiments, the ovarian cancer is primary peritoneal or fallopian tube cancer. In various embodiments, the neoplastic disease is metastatic or unresectable HGSOC, with platinum resistance defined as progression on or within 6 months of a platinum-containing regimen. In various embodiments, the ovarian cancer has been treated or is being treated with platinum-resistant relapse therapy. In various embodiments, the neoplastic disease is serous endometrial cancer. Optionally, the neoplastic disease is metastatic or recurrent serous endometrial cancer. In various embodiments, the endometrial cancer is recurrent and / or refractory to at least one line of systemic therapy in the metastatic / recurrent setting, or intolerant to existing therapy(s) known to provide clinical benefit for the neoplastic disease. In various embodiments, the neoplastic disease is an advanced or metastatic solid tumor that is unresectable, recurrent, and / or refractory or intolerant to at least one line of systemic chemotherapy. Optionally, the advanced or metastatic solid tumor is TP53. MUT It is.

[0168] In various embodiments, the cancer is ovarian cancer, breast cancer, or endometrial cancer. In various embodiments, the ovarian cancer is clear cell ovarian cancer or high grade serous ovarian cancer (HGSOC), optionally metastatic or unresectable HGSOC. Optionally, the HGSOC is platinum-resistant HGSOC, or the HGSOC has progressed during or within 6 months of a platinum-containing regimen. In various embodiments, the cancer is primary peritoneal cancer and / or fallopian tube cancer. In representative embodiments, the breast cancer is triple-negative breast cancer. In some embodiments, the subject has relapsed or is refractory to at least one line of systemic chemotherapy. Optionally, the systemic chemotherapy includes taxanes, gemcitabine, or doxorubicin. In various embodiments, the endometrial cancer is serous endometrial cancer, optionally metastatic or recurrent serous endometrial cancer. In certain embodiments, the serous endometrial cancer has relapsed or is refractory to at least one line of systemic chemotherapy, eg, cisplatin, carboplatin, or levantinib.

[0169] In various embodiments, the tumor is an advanced solid tumor. In various embodiments, the tumor is unresectable, metastatic, and / or non-localized. In representative embodiments, the tumor is recurrent or refractory to at least one line of systemic chemotherapy.

[0170] In various embodiments, the neoplastic disease is resistant to treatment with one or more drugs. In various embodiments, the neoplastic disease exhibits reduced sensitivity to treatment with one or more drugs. Optionally, the neoplastic disease is a multidrug-resistant neoplastic disease. In representative embodiments, the tumor or cancer cells (e.g., of the neoplastic disease) are multidrug-resistant tumor or cancer cells and / or exhibit increased expression of the multidrug resistance 1 (MDR-1) gene and / or its gene product. In representative embodiments, the tumor or cancer cells (e.g., of the neoplastic disease) exhibit increased expression of P-glycoprotein (P-gp), which is encoded by the MDR-1 gene. In various embodiments, the neoplastic disease exhibits reduced sensitivity or resistance to treatment with antimitotic inhibitors or anthracycline antibiotics, optionally paclitaxel or doxorubicin. In various embodiments, the tumor or cancer cells (e.g., of the neoplastic disease) exhibit mutations in the tubulin gene, overexpression of tubulin, tubulin amplification and / or isotype-switched tubulin expression. In various embodiments, mutations in α- or β-tubulin prevent the taxane from binding to the correct location on the microtubule, thereby rendering the taxane ineffective. In representative embodiments, the neoplastic disease exhibits reduced sensitivity or resistance to treatment with any one or more of platinum agents, anthracyclines, targeted therapies (e.g., TKIs, PARP inhibitors).

[0171] In various embodiments, the neoplastic disease is a cancer that contains one or more whole genome duplication or whole genome doubling (WGD) events.WGD in the context of cancer is discussed in Lens and Hemdema, Nature Reviews Cancer 19:32-45 (2019); Ganem et.al., Current Opinion in Genetics & Development 17, 157-162, and Davoli et.al., Annual Review of Cell and Developmental Biology 27, 585-610.

[0172] Inactivated genes, amplified genes and expression levels As used herein, the term "inactivated" in relation to a gene refers to the reduction or loss of function of the gene or the gene product encoded by the gene. The inactivation of a gene can be caused by one or more known mechanisms. For example, the inactivation of a gene can be caused by a mutation (e.g., including deletion) in the DNA sequence, RNA sequence, or protein sequence compared to the corresponding wild-type gene, RNA, or protein, or can be caused by epigenetic mutation without any change in the DNA sequence of the gene.

[0173] In various embodiments, the cancer cell comprises a mutation or abnormality in a gene or a gene product encoded by the gene, the mutation or abnormality being compared with the corresponding wild type gene or gene product, and the presence of the variation leads to or is associated with the silencing of the gene, the reduced or lost expression of the gene or the gene product encoded by the gene, the reduced or lost function of the gene or the gene product encoded by the gene, or a combination thereof. In various embodiments, the gene product is an RNA transcript or a protein. In various embodiments, the mutation at least leads to the reduced or lost function of the gene or the gene product encoded by the gene. In various embodiments, the mutation at least leads to the reduced or lost function of the TP53 gene or the gene product encoded by the TP53 gene. In various embodiments, the mutation at least leads to the reduced or lost function of the Rb1 gene or the gene product encoded by the Rb1 gene. In various embodiments, the mutation at least leads to the reduced or lost function of the BRCA gene or the gene product encoded by the BRCA gene.

[0174] The mutation in the gene may be present anywhere in the gene, for example, in an intron or exon, in the 5'-untranslated region (5'-UTR), or in the 3'-untranslated region (3'-UTR). The mutation may be present in or in any part of the transcript (e.g., RNA transcript, primary transcript, pre-mRNA, mRNA) encoded by the gene, or in or in any part of the protein encoded by the gene.

[0175] In various embodiments, the mutation is a difference in DNA sequence, RNA sequence or protein sequence compared to the corresponding wild type gene, RNA or protein. In various embodiments, the inactivated gene is detected by analyzing the nucleotide sequence of the gene, analyzing the nucleotide sequence of the RNA encoded by the gene, or analyzing the amino acid sequence of the protein encoded by the gene, and comparing the sequence of the gene of the sample with the corresponding wild type human sequence of the gene, RNA or protein. In representative embodiments, the mutation comprises one or more nucleotide deletions, insertions or substitutions in the DNA sequence or RNA sequence, one or more amino acid deletions, insertions or substitutions in the protein sequence, compared to the corresponding wild type gene, RNA or protein. In representative embodiments, the mutation comprises one or more nucleotide deletions, insertions or substitutions in the DNA sequence or RNA sequence, one or more amino acid deletions, insertions or substitutions in the protein sequence, compared to the corresponding wild type gene, RNA or protein, which may result in an increase in gene copy number or amplification of the DNA, RNA or protein. In various embodiments, the cancer cell comprises a genetic mutation in the gene. In various embodiments, the cancer cells comprise a genetic mutation in a gene, or a deletion of a nucleotide in a gene. In representative embodiments, the genetic mutation is a missense mutation, a nonsense mutation, an insertion, a deletion, a duplication, a frameshift mutation, a truncation, or a repeat expansion. In various embodiments, the inactivated TP53 gene comprises a mutation, a deletion, or a truncation, the inactivated Rb1 gene comprises a mutation, a deletion, or a truncation, and / or the inactivated BRCA gene comprises a mutation, a deletion, or a truncation. As used herein, the term "BRCA gene" refers to the BRCA1 or BRCA2 gene. In representative embodiments, the BRCA gene is BRCA1. In representative embodiments, the BRCA gene is BRCA2.

[0176] In various embodiments, the variation is epigenetic and does not include any change in the DNA sequence of the gene. In a representative embodiment, the inactivated gene is epigenetically silenced, and optionally includes covalent modification of DNA or histone protein. The covalent modification of DNA can be, for example, cytosine methylation or hydroxymethylation. The covalent modification of histone protein can be, for example, lysine acetylation, lysine or arginine methylation, serine or threonine phosphorylation, or lysine ubiquitination or sumoylation. The mechanism of gene silencing can occur during transcription or translation. Representative mechanisms of gene silencing include, but are not limited to, DNA methylation, histone modification, and RNA interference (RNAi). In various embodiments, the inactivated gene is an epigenetically silenced gene with an epigenetically silenced promoter. Optionally, the inactivated TP53 gene has an epigenetically silenced TP53 promoter, or the inactivated Rb1 gene has an epigenetically silenced Rb1 promoter, or the inactivated BRCA gene has an epigenetically silenced BRCA promoter.Suitable techniques for assaying epigenetic silencing include, but are not limited to, chromatin immunoprecipitation (ChIP-on chip, ChIP-Seq), fluorescent in situ hybridization (FISH), methylation-sensitive restriction enzyme, DNA adenine methyltransferase identification (DamID), and bisulfite sequencing.See, for example, Verma et.al., Cancer Epidemiology, Biomarkers, and Prevention 23:223-233 (2014).

[0177] In various embodiments, the inactivated gene is inactivated by virus-induced gene silencing (VIGS).In various embodiments, the inactivated TP53 gene is inactivated by viral protein, for example, human papillomavirus (HPV) E6 protein.Optionally, HPV E6 protein interacts with p53 protein encoded by TP53 gene, and inactivates p53 protein.In various embodiments, the inactivated Rb1 gene is inactivated by viral protein, for example, HPV E7 protein.Optionally, HPV E7 protein interacts with Rb protein encoded by Rb1 gene, and inactivates Rb protein.Such silencing methods are known in the art.See, for example, Jiang and Milner, Oncogene 21:6041-6048 (2002).

[0178] In various embodiments of the disclosed methods, the cancer cells contain gene amplification, e.g., CCNE1 amplification, or an increase in gene copy number, e.g., a gene copy number increase of a gene. In various embodiments, the cancer cells contain gene copy number increase or amplified genes that can be detected by DNA or RNA-based techniques (gene expression analysis [comparative genomic hybridization, RNA-based hybridization], NGS, PCR, or Southern blot) or by molecular cytogenetic techniques (FISH2, CISH (chromogenic in situ hybridization) using gene-specific probes. In various embodiments, competitive or quantitative PCR, genomic hybridization to cDNA microarrays, and hybridization and quantification of gene probes to RNA are performed to detect gene amplification or gene copy number increase. See, e.g., Harlow and Stewart, Genome Res 3:163-168 (1993); Heiskanen et.al., Cancer Res 60(4):799-802(2000). In various embodiments, the cancer cells comprise a gene copy number increase or amplification of the MDM2 gene, and / or a gene copy number increase or amplification or mutation of the FBXW7 gene. In a representative embodiment, the cancer cells comprise a gene copy number increase or amplification of the MDM2 gene, and a decrease in p53 protein level. In a representative embodiment, the cancer cells comprise a mutation of the FBXW7 gene, and an overexpression of the gene product encoded by the CCNE1 gene. Next generation sequencing (NGS) may also be used as a method for detecting gene copy number increase or decrease or gene amplification, whereby a gene region is sequenced, and the sequencing reads are compared with other genes to infer the increase or decrease of the gene of interest.

[0179] In a representative embodiment, the inactivated TP53 gene comprises (i) a TP53 gene mutation, deletion, truncation and / or epigenetically silenced TP53 promoter, (ii) inactivated by a viral protein or via gene amplification of the MDM2 gene, or (iii) a combination thereof. Optionally, the viral protein is a human papillomavirus (HPV) E6 protein. In a representative embodiment, the inactivated Rb1 gene comprises (i) an Rb1 gene mutation, deletion, truncation and / or epigenetically silenced Rb1 promoter, (ii) inactivated by a viral protein, or (iii) a combination thereof. Optionally, the viral protein is a human papillomavirus (HPV) E7 protein. In a representative embodiment, the inactivated BRCA gene comprises (i) a BRCA gene mutation, deletion, truncation and / or epigenetically silenced BRCA promoter. Optionally, the BRCA gene is a BRCA1 gene. Alternatively, the BRCA gene is the BRCA2 gene.

[0180] In various embodiments, the inactivated TP53 gene, the inactivated Rb1 gene, the CCNE1 gene copy number gain or amplification, and / or the inactivated BRCA gene are present in the germline cells of the neoplastic disease (e.g., cancer). In various embodiments, the inactivated TP53 gene, the inactivated Rb1 gene, the CCNE1 gene copy number gain or amplification, and / or the inactivated BRCA gene are present in the germline cells of the neoplastic disease (e.g., cancer) and are not present in the somatic cells of the neoplastic disease (e.g., cancer). Optionally, due to somatic mutations in the neoplastic disease, the somatic cells of the neoplastic disease have reverted to a wild-type genotype and therefore do not exhibit the inactivated TP53 gene, the inactivated Rb1 gene, the CCNE1 gene copy number gain or amplification, and / or the inactivated BRCA gene, while the germline cells of the neoplastic disease still exhibit the inactivated TP53 gene, the inactivated Rb1 gene, the CCNE1 gene copy number gain or amplification, and / or the inactivated BRCA gene. For example, the neoplastic disease can be a PARP inhibitor-resistant cancer, where only germline cells of the cancer have an inactivated BRCA1 gene, while somatic cells of the cancer show restoration of the BRCA1 coding region and function.

[0181] Cytogenetic and / or molecular methods can be used to detect the presence of inactivated or amplified genes or gene copy number increase, such as inactivated TP53 gene, inactivated Rb1 gene, amplified CCNE1 gene or inactivated BRCA gene. In a representative embodiment, direct DNA sequencing, DNA hybridization and / or restriction enzyme digestion are used. Optionally, the cytogenetic method includes karyotyping, fluorescence in situ hybridization (FISH), comparative genomic hybridization (CGH), or a combination thereof. In various embodiments, the molecular method includes restriction fragment length polymorphism (RFLP), amplification resistant mutation system (ARMS), polymerase chain reaction (PCR), multiplex ligation-dependent probe amplification (MLPA), denaturing gradient gel electrophoresis (DGGE), single-strand conformation polymorphism (SSCP), heteroduplex analysis, chemical cleavage of mismatches (CCM), protein truncation test (PTT), oligonucleotide ligation assay (OLA), or a combination thereof. Optionally, the PCR is multiplex PCR, nested PCR, RT-PCR or real-time quantitative PCR. In various embodiments, the expression level of RNA or protein encoded by TP53, Rb1, CCNE1 and / or BRCA genes is assayed. In various embodiments, ARMS, FISH, IHC or NGS is employed. Such techniques are described in Su et al., J Experimental Clin Cancer Research 36:121(2017) and He et al., Blood 127(24):3004-3014(2016). In various embodiments, whole exome or whole genome sequencing is used. In a representative embodiment, the assay comprises liquid biopsy. Liquid biopsy has been described in detail in the art. See, for example, Poulet et al., Acta Cytol 63(6):449-455(2019); Chen and Zhao, Hum Genomics 13(1):34(2019).

[0182] In various embodiments, the gene copy number increase or amplification leads to overexpression or elevated levels of the gene product (e.g., RNA and / or protein) encoded by the gene. Methods for detecting elevated RNA and / or protein levels are known in the art. In a representative embodiment, the gene copy number increase or amplification of the CCNE1 gene leads to overexpression or elevated levels of the gene product encoded by the CCNE1 gene. In a representative embodiment, the overexpression of the CCNE1 gene product is caused by a mutation in the FBXW7 gene. In various embodiments, the sample is positive for overexpression of the CCNE1 gene product and a mutation in the FBXW7 gene.

[0183] Suitable methods for determining the expression level of nucleic acids (e.g., genes, RNA, mRNA) are known in the art and include, but are not limited to, quantitative polymerase chain reaction (qPCR) (e.g., quantitative real-time PCR (qRT-PCR)), RNAseq, nanostring, and northern blotting. Techniques for measuring gene expression also include gene expression assays with or without gene chips, or gene expression microarrays, as described, for example, in Onken et.al., J Molec Diag 12(4):461-468(2010) and Kirby et.al., Adv Clin Chem 44:247-292(2007). Affymetrix gene chips and RNA chips and gene expression assay kits (e.g., Applied Biosystems™ TaqMan® Gene Expression Assays) are also commercially available from companies such as ThermoFisher Scientific (Waltham, MA), and Nanostring (Geiss et.al., Nature Biotechnology 26:317-325(2008)). Suitable methods for determining protein expression levels are known in the art and include immunoassays (e.g., Western blotting, enzyme-linked immunosorbent assays (ELISAs), radioimmunoassays (RIAs), and immunohistochemistry assays), or bead-based multiplex assays, such as those described in Djoba Siawaya JF, Roberts T, Babb C, Black G, Golakai HJ, Stanley K, et al. (2008) An Evaluation of Commercial Fluorescent Bead-Based Luminex Cytokine Assays. PLoS ONE 3(7):e2535. Proteomic analysis, the systematic identification and quantification of proteins in a particular biological system, is known and mass spectrometry is commonly used for this purpose.

[0184] In a representative embodiment, the method includes measuring the level of complementary DNA (cDNA) based on the RNA encoded by the gene. Briefly, the method includes extracting or isolating RNA from the sample (e.g., from the tumor cell(s) of the sample) and synthesizing cDNA based on the RNA isolated from the sample. Alternatively, or in addition, in some embodiments, measuring the expression level includes isolating RNA from the sample, generating complementary DNA (cDNA) from the RNA, amplifying the cDNA, and hybridizing the cDNA to a gene expression microarray. Thus, in some embodiments, measuring the expression level includes isolating RNA from the sample and quantifying the RNA by RNA-Seq. In alternative or further embodiments, the expression level is determined via an immunohistochemistry assay. In a representative embodiment, measuring the expression level includes contacting the sample with a binding agent against TP53, Rb1, BRCA, or CCNE1, or a gene product thereof, or a combination thereof. In some embodiments, the binding agent is an antibody or an antigen-binding fragment thereof. In some embodiments, the binding agent is a nucleic acid probe specific for TP53, Rb1, BRCA or CCNE1, or an RNA transcript thereof, or a complement thereof.

[0185] Once the expression level of TP53, Rb1, BRCA or CCNE1, or its gene product, is measured from a sample obtained from a subject, the measured expression level can be compared to a reference level, normalized to a housekeeping gene, and mathematically transformed.In a representative embodiment, the measured expression level of TP53, Rb1, BRCA or CCNE1, or its gene product, is centered and scaled.Suitable techniques for centering and scaling biological data are known in the art.See, for example, van den Berg et.al., BMC Genomics 7:142(2006).

[0186] Wild-type TP53, Rb1, CCNE1 and BRCA genes, as well as the RNAs and proteins encoded by these genes, are known in the art, and representative sequences for each are available on the National Center for Biotechnology Information (NCBI) website and are provided in the Sequence Listing submitted herewith. [Table 3]

[0187] Cancer cells may be identified as "positive" or "negative" for at least one of: (a) an inactivated TP53 gene, and / or (b) (i) an inactivated Rb1 gene, (ii) an amplified CCNE1 gene, an increase in gene copy number of the CCNE1 gene, or overexpression of the CCNE1 gene product, (iii) an inactivated BRCA gene, or (iv) a combination thereof. As used herein, the term "positive" in relation to a sample means that at least one of: an inactivated TP53 gene, and / or (b) (i) an inactivated Rb1 gene, (ii) an amplified CCNE1 gene, an increase in gene copy number of the CCNE1 gene, or overexpression of the CCNE1 gene product, (iii) an inactivated BRCA gene, or (iv) a combination thereof is present in the sample. As used herein, the term "negative" in relation to a sample means that at least one of the following is not present in the sample: an inactivated TP53 gene, and / or (b)(i) an inactivated Rb1 gene, (ii) an amplified CCNE1 gene, an increase in gene copy number of the CCNE1 gene or overexpression of the CCNE1 gene product, (iii) an inactivated BRCA gene, or (iv) a combination thereof, e.g., the sample does not have an inactivated TP53 gene, and / or at least one of the following is present in the sample: (b)(i) an inactivated Rb1 gene, (ii) an amplified CCNE1 gene, an increase in gene copy number of the CCNE1 gene or overexpression of the CCNE1 gene product, (iii) an inactivated BRCA gene, or (iv) a combination thereof.

[0188] subject In representative embodiments of the present disclosure, the subject is a mammal, including, but not limited to, mammals of the order Rodentia, such as mice and hamsters, and mammals of the order Lagomorpha, such as rabbits, mammals of the order Carnivora, including Felidae (cats) and Canidae (dogs), mammals of the order Artiodactyla, including Bovidae (cows) and Suidae (pigs), or mammals of the order Perissodactyla, including Equidae (horses). In some aspects, the mammal is a mammal of the order Primates, Ceboids, or Simoids (monkeys), or apes (humans and apes). In some aspects, the mammal is a human. In various aspects, the subject has a neoplastic disease, such as any one of those described herein. As used herein, the terms "patient," "subject," or "mammal" refer to any "patient," "subject," or "mammal," including humans, cows, horses, dogs, and cats. In one embodiment of the present invention, the mammal is a human. In various aspects, the subject is an adult human. Optionally, the subject has been pretreated with at least one chemotherapeutic agent.

[0189] In a representative embodiment, the subject has a cancer, including metastatic cancer, unresectable tumor, or a combination thereof. In various embodiments, the cancer or tumor shows or has shown resistance or reduced sensitivity to treatment with a CDK4 / 6 inhibitor. In a representative embodiment, the subject has breast cancer, optionally luminal breast cancer or triple-negative breast cancer (TNBC). In various embodiments, the breast cancer is (a) histologically or cytologically confirmed to be metastatic or locally recurrent estrogen receptor (ER) negative (e.g., <1% by immunohistochemistry [IHC]), (b) progesterone receptor (PR) negative (e.g., <1% IHC), and (c) human epidermal growth factor receptor 2 (Her2) negative (either fluorescent in situ hybridization [FISH] negative, 0 or 1+ by IHC, or IHC 2+ and FISH negative, according to ASCO / CAP definition). In representative embodiments, the subject is relapsed and / or refractory to at least one line of systemic chemotherapy in the metastatic setting or intolerant to existing treatment(s) known to provide clinical benefit for those conditions. In representative embodiments, the subject has been previously exposed to an immune checkpoint inhibitor. In various embodiments, the breast cancer is hormone receptor (HR) positive and / or HER2 negative. In various embodiments, the breast cancer is advanced and / or metastatic breast cancer. In various embodiments, the subject has HR+ / HER2- advanced or metastatic breast cancer that has progressed after receiving endocrine therapy. In some embodiments, the subject is a patient with hormone receptor positive (HR+) / HER2 negative (HER2-) advanced or metastatic breast cancer who was previously treated with endocrine therapy and chemotherapy after the cancer had spread / metastasized. In various embodiments, the subject has HR+ / HER2- advanced or metastatic breast cancer in postmenopausal women who have not been previously treated with hormone therapy (Arimidex (chemical name: anastrozole), Aromasin (chemical name: exemestane), and Femara (chemical name: letrozole). In various embodiments, the subject is a postmenopausal woman with HR+ / HER2- advanced or metastatic breast cancer that has grown after treatment with hormone therapy.In certain embodiments, the subject is a pre- / peri- or post-menopausal woman with HR+, human epidermal growth factor receptor 2 (HER2) negative advanced or metastatic breast cancer and has undergone endocrine-based therapy. Optionally, the subject is a post-menopausal woman with HR+, HER2- advanced or metastatic breast cancer and has undergone initial endocrine-based therapy or has disease progression during treatment with endocrine therapy. In various embodiments, the subject has ovarian cancer, optionally high-grade serous ovarian cancer (HGSOC). Optionally, the ovarian cancer is platinum-resistant HGSOC. In representative embodiments, the subject has primary peritoneal and / or fallopian tube cancer. In various embodiments, the subject has a histologically or cytologically confirmed diagnosis of metastatic or unresectable HGSOC that is platinum-resistant, defined as progressive, during or within 6 months of a platinum-containing regimen. In various embodiments, the subject has ovarian cancer and has undergone or is undergoing platinum-resistant relapse therapy. In various aspects, the subject has serous endometrial cancer. Optionally, the subject has a histologically or cytologically confirmed diagnosis of metastatic or recurrent serous endometrial cancer. In various embodiments, the subject is recurrent and / or refractory to at least one line of systemic therapy in the metastatic / recurrent setting, or intolerant to existing treatment(s) known to provide clinical benefit for these conditions. In various embodiments, the subject has an advanced or metastatic solid tumor that is unresectable, recurrent, and / or refractory or intolerant to at least one line of systemic therapy. Optionally, the advanced or metastatic solid tumor is TP53. MUT It is.

[0190] In representative embodiments, the patient does not have any of the following: (a) active brain metastases; (b) primary central nervous system (CNS) tumors, hematological malignancies or lymphomas; (c) uncontrolled pleural effusion, pericardial effusion, or ascites; (d) gastrointestinal (GI) tract disease precluding oral medication.

[0191] Subject selection and treatment outcomes In some embodiments, the subject treated by compound A in the disclosed methods has undergone one or more prior systemic cancer therapies (e.g., compound A is a second-line or third-line treatment). The prior systemic cancer therapy can be any therapy approved by a regulatory agency (e.g., FDA or EMA) as a treatment for a specific type and stage of cancer. In some cases, the prior systemic cancer therapy is a cancer treatment that has not yet been approved by a regulatory agency but is in clinical trials. If the subject has previously undergone systemic cancer therapy, in some cases, the subject has not undergone any systemic cancer therapy for at least 1 month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, or at least 6 months before starting treatment with compound A disclosed herein.

[0192] Subjects undergoing treatment are monitored for adverse events (AEs) during the treatment period. Treatment-related AEs are AEs related to the therapeutic agent. Treatment-emergent AEs are adverse events that a subject develops during treatment that were not present before treatment began. In some cases, treatment-emergent AEs are not related to or suspected to be not related to the treatment itself. AEs are characterized by one of five grades, with grade 1 being mild AEs, grade 2 being moderate AEs, grade 3 being severe AEs, grade 4 being life-threatening or disabling AEs, and grade 5 being death related to an AE. In some cases, the subject does not exhibit any treatment-related grade 3 AEs. In some cases, the subject does not exhibit any grade 3 AEs. In some cases, the subject does not exhibit any treatment-related grade 4 AEs. In some cases, the subject does not exhibit any grade 4 AEs. In various embodiments, the subject does not exhibit any treatment-related grade 3 or grade 4 AEs for at least one month or at least three months after administration of Compound A.

[0193] In various cases, subjects treated with Compound A in the methods disclosed herein do not exhibit any dose-limiting toxicity (DLT) at the dose administered. DLT refers to any AE that occurs during the first treatment cycle (days 1 to 21) of Compound A and meets the criteria listed below that cannot be ruled out as being related to the drug. AE grading is based on the CTCAE version 5.0 guidelines. AEs for DLT evaluation: Hematological toxicity: febrile neutropenia; neutropenic infection; grade 4 neutropenia; grade 3 or higher thrombocytopenia for more than 7 days; grade 3 thrombocytopenia with grade 2 or higher bleeding; grade 4 thrombocytopenia; grade 4 anemia, grade 4 or higher non-hematologic toxicity, vomiting or diarrhea; grade 3 diarrhea, or grade 3 vomiting lasting more than 3 days despite optimal medical support; grade 3 or higher nausea lasting 3 days or more despite optimal medical support; any other grade 3 or higher AE.

[0194] In various embodiments, the subject of the disclosed method shows a response to treatment. In some cases, the subject shows at least stable disease (SD) upon administration of compound A. In some cases, the subject shows at least partial response (PR) upon administration of compound A. The subject's response is evaluated by the criteria defined in RECIST 1.1, for example, as discussed in Eisenhauer et al., Eur J Cancer, 45:228-247 (2009). A complete response (CR) is the disappearance of all target lesions and the short axis of any pathological lymph node is reduced to less than 10 mm. A partial response (PR) is a reduction in the sum of lesion diameters of target lesions by at least 30% based on the sum of lesion diameters at baseline. Progressive disease is an increase in the sum of the diameters of target lesions of at least 20%, with an absolute increase of at least 5 mm based on the minimum sum during the study (including the baseline sum if it is the minimum sum during the study). Stable disease is neither sufficient shrinkage to qualify for PR nor sufficient growth to qualify for PD. Disease control is when patients may alternate between stable disease and partial responses. Tumor size can be measured by radiography.

[0195] Other embodiments Further provided herein are the hydrochloride salt (Compound A-HCl), mesylate salt (Compound A-MsA), tosylate salt (Compound A-TsA), sulfate salt (Compound A-Sulfate), variable hydrate (Compound A-variable hydrate), tetrahydrofuran solvate (Compound A-THF), ethanol solvate (Compound A-Ethanol), 1-propanol solvate (Compound A-1-Propanol), isopropyl alcohol solvate (Compound A-IPA), methanol solvate (Compound A-Methanol), isopropyl acetate solvate (Compound A-IPAc), acetone solvate (Compound A-Acetone), cyclopentyl methyl ether solvate (Compound A-CPME), dioxane solvate (Compound A-Dioxane), ethyl acetate solvate (Compound A-EtOAc), acetonitrile solvate (Compound A-MeCN), methyl tert-butyl ether solvate (Compound A-MTBE). a toluene solvate (Compound A-toluene) dodecyl sulfate (Compound A-dodecyl sulfate), a dimethylformamide (DMF) solvate hydrate (Compound A-DMF-hydrate), a dimethylacetamide (DMAC) solvate (Compound A-DMAC), a monobesylate hydrate (Compound A-besylate-hydrate), a caffeine co-crystal (Compound A-caffeine), a citric acid co-crystal (Compound A-citric acid), a saccharin co-crystal (Compound A-saccharin), an L-tartaric acid co-crystal (Compound A-AL-tartaric acid), or a urea co-crystal (Compound A-urea); and a pharmaceutical composition comprising any of the salts, solvates, or co-crystals of Compound A, and a pharma- ceutical composition comprising a pharma- ...

[0196] In various embodiments, the organic solvent may be selected from the group consisting of an ether solvent, a non-polar solvent, and any combination thereof. In some cases, the organic solvent may be an ether solvent. Suitable ether solvents may include, for example, tetrahydrofuran (THF), 2-methyltetrahydrofuran (MeTHF), cyclopentyl methyl ether, tert-butyl methyl ether, 1,2-dimethoxyethane, 1,4-dioxane, diethyl ether, diisopropyl ether, bis(2-methoxyethyl) ether, propylene glycol methyl ether, and any combination thereof. In embodiments, the ether solvent may be THF or 2-methyltetrahydrofuran. In some cases, the organic solvent may be a non-polar solvent. Suitable non-polar solvents may include, for example, hexane, pentane, toluene, benzene, heptane, xylene, and any combination thereof. In embodiments, the non-polar solvent may be toluene, hexane, heptane, or any combination thereof. In embodiments, the organic solvent may be selected from the group consisting of THF, 2-methyltetrahydrofuran, cyclopentyl methyl ether, tert-butyl methyl ether, 1,2-dimethoxyethane, toluene, hexane, heptane, 1,4-dioxane, and any combination thereof. In some embodiments, the organic solvent is THF.

[0197] This disclosure should be read in conjunction with its detailed description, with the understanding that the foregoing description is intended to be illustrative, and not limiting, of the scope of the disclosure, which is defined by the scope of the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims. For example, as shown in Examples 1-45. EXAMPLES

[0198] The following examples are offered for illustrative purposes and are not intended to limit the scope of the invention.

[0199] Materials and Methods Commercially available reagents were used as is without further purification unless otherwise specified. 1.0 M MeI in THF was prepared by weight. Batch and flow chemistry equipment (reactors, tubing, pumps, connections, fittings) were used from commercially available sources.

[0200] The synthesis of the starting material (Compound A) for the following synthetic method is disclosed in U.S. Nonprovisional Patent Application No. 16 / 724,119, filed December 20, 2019, and published July 30, 2020 as US2020-0239441. Starting materials, intermediates, and end products of reactions may be isolated and purified, if desired, using conventional techniques, including, but not limited to, filtration, distillation, crystallization, chromatography, and the like. Such materials may be characterized using conventional means, including physical constants and spectral data.

[0201] Unless otherwise specified, the reactions described herein are conducted at atmospheric pressure and at temperatures ranging from about -78°C to about 150°C, or from about 0°C to about 50°C, or from about 15°C to about 25°C.

[0202] PANalytical X'Pert PRO MPD Diffractometer - Transmission Geometry Unless otherwise specified, XRPD patterns were collected on a PANalytical X'Pert PRO MPD diffractometer with Cu radiation generated by an Optix long fine focus source as the incident beam. An elliptically graded multilayer mirror was used to focus the Cu Kα X-rays through the sample onto the detector. Prior to analysis, silicon samples (NIST SRM 640f) were analyzed to confirm the Si 111 peak position. Sample specimens were sandwiched between 3 μm thick films and analyzed in transmission geometry. A beam stop and short anti-scatter extensions were used to minimize background from air. Soller slits were used for the incident and diffracted beams to minimize broadening due to axial divergence. Diffraction patterns were collected using a scanning position sensitive detector (X'Celerator) positioned 240 mm from the sample and Data Collector software v.5.5 (except for materials as received, for which Data Collector software v.2.2b was used). The data acquisition parameters for each pattern are displayed above the images in the data section of this report.

[0203] PANalytical X'Pert PRO MPD Diffractometer - Reflection Geometry Unless otherwise specified, XRPD patterns were collected on a PANalytical X'Pert PRO MPD diffractometer using a long, finely focused source and a CuKα incident beam generated using a nickel filter. The diffractometer was configured using a symmetric Bragg-Brentano geometry. Data were collected and analyzed using Data Collector software v.2.2b. Prior to analysis, silicon samples (NIST SRM 640f) were analyzed to ensure that the observed position of the Si 111 peak was consistent with the NIST certified position. Sample specimens were packed into nickel-coated copper wells. Anti-scatter slits (SS) were used to minimize background from air. Soller slits were used for the incident and diffracted beams to minimize broadening due to axial divergence. Diffraction patterns were collected using a scanning position-sensitive detector (X'Celerator) positioned 240 mm from the sample and Data Collector software v.2.2b. The data acquisition parameters for each pattern, including the divergence slit (DS) and the incident beam anti-scatter slit (SS), are displayed above the images in the data section of this report.

[0204] X-ray powder diffraction (XRPD) data were acquired on a PANalytical X'Pert PRO X-ray diffraction system equipped with a RTMS detector. Samples were scanned at ambient temperature using CuKα radiation (1.541874 Å) at 45 kV, 40 mA in continuous mode from 5° to 45° (2θ) with a step size of 0.0334° and a time of 50 seconds per step.

[0205] XRPD indexing was performed using SSCI's proprietary software TRIADS™, as described in US Pat. No. 8,576,985.

[0206] Differential scanning calorimetry (DSC) was performed using a Mettler-Toledo DSC3+ differential scanning calorimeter. Taurag adjustments are made with indium, tin, and zinc. Temperature and enthalpy adjustments are made with octane, phenyl salicylate, indium, tin, and zinc. The adjustments are then verified with octane, phenyl salicylate, indium, tin, and zinc. Samples were placed into sealed aluminum DSC pans and the weights were accurately recorded. The lids of the pans were pierced by the instrument and inserted into the DSC cell for analysis. A weighing aluminum pan configured as the sample pan was placed on the reference side of the cell.

[0207] Alternatively, differential scanning calorimetry (DSC) analyses were performed using TA Instruments Q and Discovery series calorimeters under 50 ml / min dry nitrogen in aluminum pans from 25° C. to 250° C. to 350° C. at 10° C. / min.

[0208] Thermogravimetric analysis (TGA) and combined TGA / DSC analysis were performed using a Mettler-Toledo TGA / DSC3+ analyzer. Temperature and enthalpy adjustments were made with indium, tin, and zinc, then probed with indium. The balance was probed with calcium oxalate. Samples were placed in open aluminum pans. The pans were sealed, holes were drilled in the lid, and inserted into the TG furnace. The weighed aluminum pan configured as the sample pan was placed on the reference platform. The furnace was heated under nitrogen.

[0209] Thermogravimetric analysis (TGA) was performed using TA Instruments Q and Discovery series analyzers in platinum pans under 25 ml / min dry nitrogen from ambient temperature to between 250° C. and 350° C. at 10° C. / min.

[0210] Moisture sorption data was collected using a VTI SGA 100 Symmetrical Vapor Sorption Analyzer. Approximately 5 mg to 10 mg of sample was placed in platinum pans. Hygroscopicity was evaluated from 5% RH to 95% RH in 5% RH increments. Adsorption and desorption cycles were collected. Equilibrium criteria were 0.001% weight change in 10 minutes with a maximum equilibration time of 180 minutes.

[0211] Solution proton NMR spectra were acquired at 25° C. on a Varian UNITYINOVA-400 spectrometer by Spectral Data Services of Champaign (SSCI), IL. Samples were dissolved in DMSO-d6. In some cases, solution NMR spectra were also acquired at SSCI on an Agilent DD2-400 spectrometer using deuterated DMSO or methanol.

[0212] 19 F SSNMR data is from 600 MHz ( 1 H. A 4 mm H / F / X spinning probe operating at a spinning frequency of 14 kHz was used for all experiments. The HPDEC program was used with a 10 s recycle delay and referenced to Teflon. 1 H90° pulse and 5μs 19 An F 90° pulse was used. Decoupling was performed using the spinal64 sequence. 256 transients were acquired for signal averaging. Data were processed with Topspin 3.0 software.

[0213] Example 1: Crystalline Compound A-HCl Form 1 Crystallization of compound A-HCl form 1 can be achieved in multiple solvent systems after in situ protonation of compound A with hydrochloric acid. Initially, compound A-HCl form 1 was prepared by slurrying 1 equivalent of hydrochloric acid in acetonitrile / water 90 / 10 at ambient conditions. An anhydrous process was then used in which compound A was treated with hot hydrochloric acid in an acetonitrile / 1,4-dioxane system (Table 1, entry no. 1). Alternative reactive crystallization processes using different hydrochloric acid sources were developed in NMP / EtOH, THF / water, and acetone / water (Table 1, entries no. 2-5). Acetone / water was selected as the final crystallization system due to the consistently high purity of the drug substance and the ability to control residual solvent content according to ICH guideline limits. Characterization results of these batches are summarized in Table 1.

[0214] [Table 4]

[0215] Compound A was dissolved in 30 volumes of acetone, followed by polish filtration and the addition of 5 volumes of water and 2.0 equivalents of hydrochloric acid (2.5 volumes of 1.5 N aqueous hydrochloric acid) at ambient temperature. The final solvent composition for crystallization and slurry aging was 80 / 20 (v / v) acetone / water, which provided adequate solubility for both Compound A (i.e., approximately 18 mg / mL) and Compound A-HCl Form 1 (i.e., approximately 8 mg / mL) to achieve crystal growth and impurity rejection. The process was performed unseeded, with crystal growth occurring during the addition of hydrochloric acid to the Compound A solution. The final slurry was aged at ambient temperature for 10 hours and cooled to 10°C prior to wet-grinding. The grind was isolated at 10°C, followed by washing the filter cake with 8 volumes of acetone. The material was dried under vacuum at 40°C. Wet-grinding experiments in both THF / water and acetone / water resulted in particle size reduction to the specified target ranges, as outlined in Table 2, with shape purity of 95% or greater by XRPD, solid-state NMR, and DSC.

[0216] [Table 5]

[0217] Powder X-ray diffraction: Powder X-ray diffraction data were acquired on a PANalytical X'Pert PRO X-ray diffraction system with a RTMS detector. Samples were scanned from 5° to 45° (2θ) at 45 kV and 40 mA in continuous mode with a step size of 0.0334°. The incident beam path was equipped with a 0.02 rad Soller slit, a 15 mm mask, a 4° fixed anti-scatter slit, and a programmable divergence slit. The diffracted beam was equipped with a 0.02 rad Soller slit, a programmable anti-scatter slit, and a 0.02 mm nickel filter. Samples were prepared in a low background sample holder and placed on a rotating stage with a rotation time of 2 s. For variable temperature testing, samples were prepared in a flat sample holder and placed on a TTK-450 temperature control stage. For variable humidity testing, a modular humidity generator (ProUmid) was used to control the atmosphere in the THC humidity sample chamber. The XRPD pattern of the crystalline Compound A-HCl Form 1 material is shown in FIG. 1 and the XRPD peaks are shown in Table 3.

[0218] [Table 6-1] [Table 6-2]

[0219] [Table 7]

[0220] Thermal Analysis: Differential scanning calorimetry (DSC) was performed in aluminum Tzero pans using a TA Instruments Q1000 / 2000 calorimeter under dry nitrogen at a flow rate of 50 ml / min. Thermogravimetric analysis (TGA) was performed in platinum pans using a TA Instruments Q500 analyzer under dry nitrogen at 60 ml / min. The DSC and TGA of crystalline Compound A-HCl Form 1 are shown in Figure 2. Typical DSC and TGA of crystalline Compound A-HCl Form 1 showed an onset melting temperature of 271.5°C and a weight loss of about 4% before melting and decomposition.

[0221] Dynamic Vapor Sorption (DVS): Moisture sorption data was collected using a Surface Measurement Systems DVSAdvantage instrument. Equilibration criteria were ±0.001% weight change over 10 minutes with a maximum equilibration time of 360 minutes. The moisture sorption profile of crystalline Compound A-HCl Form 1 is shown in Figure 3. A typical DVS of crystalline Compound A-HCl Form 1 showed less than about 0.5% weight gain at 95% RH.

[0222] Single Crystal Data: Single crystals of crystalline Compound A-HCl Form 1 were grown at room temperature from DMF, DMAC or NMP by adding excess hydrochloric acid. Single, colorless, needle-like crystals were used for the single crystal structure determination of Compound A-HCl Form 1. Specimens selected for data collection had approximate dimensions of 0.29 × 0.08 × 0.06 mm. 3The crystal was attached to a nylon loop lubricated with paraton oil and measured on a Bruker APEX-II CCD diffractometer. The crystal was kept stable at T = 173(2) K during data collection. The structure was solved with the ShelXT (Sheldrick, GM (2015). Acta Cryst. A71, 3-8) structure solving program using the Intrinsic Phasing solver method, with Olex2 (Dolomanov et al., 2009) as the graphical interface. The model was refined with ShelXL (Sheldrick, Acta Cryst. A64 2008, 112-122) version 2018 / 3, using the least squares method. Table 5 shows a summary of the crystallographic data for crystalline compound A-HCl form 1. The molecular structure of crystalline compound A-HCl form 1 obtained from the X-ray crystal structure determination is shown in Figure 5.

[0223] [Table 8]

[0224] Example 2: Crystalline Compound A-HCl Form 2 Crystalline Compound A-HCl Form 2 was produced under high-throughput slurry conditions using 1 equivalent of HCl in a 90 / 10 acetone / water solvent. This metastable form had a low melting point and could not be scaled up or reproduced.

[0225] Powder X-ray Diffraction: The XRPD pattern of the crystalline Compound A-HCl Form 2 material is shown in FIG.

[0226] Thermal Analysis: The DSC of crystalline Compound A-HCl Form 2 is shown in Figure 8. A typical DSC of crystalline Compound A-HCl Form 2 showed an onset of melting at 113.2°C.

[0227] Example 3: Amorphous Compound A-HCl Amorphous compound A-HCl was isolated by rotary evaporation from a methanol solution and showed X-ray amorphous properties with broad peak(s). The glass transition temperature (T g) was 124°C as shown by modulated differential scanning calorimetry (MDSC) (Figure 9). The compound converted to crystalline Compound A-HCl Form 1 when heated at 165°C to 180°C. The compound converted to crystalline Compound A-HCl Form 1 and Compound A hydrate Form 2 when stressed with water.

[0228] Example 4: Crystalline Compound A-MsA Form 1 Crystalline Compound A-MsA Form 1 was prepared by slurrying Compound A with 1 molar equivalent of methanesulfonic acid in acetonitrile at ambient conditions. Gram levels were prepared on a larger scale by dissolving 3 g of Compound A in ethyl acetate (30 ml) at 60° C. in a Mettler Toledo EasyMax controlled laboratory reactor equipped with an overhead stirrer. One molar equivalent of methanesulfonic acid (350 μl) was added and precipitation was observed. The slurry was aged at 60° C. for 8 hours and cooled to 20° C. at 0.1° C. / min. After aging overnight at 20° C., the solid was isolated by vacuum filtration. The wet cake was washed with ethyl acetate (15 ml). XRPD analysis showed that the wet cake was Compound A-MsA Form 1. The wet cake was then vacuum dried at ambient temperature for 4 days and characterized. The yield is 89%.

[0229] Powder X-ray diffraction: The XRPD pattern of the crystalline Compound A-MsA Form 1 material is shown in FIG. 10 and the XRPD peaks are listed in Table 6.

[0230] [Table 9-1] [Table 9-2]

[0231] Indexing Solution of Crystalline Compound A-MsA Form 1: XRPD indexing is a method that can be used to extract information from and aid in the interpretation of an XRPD pattern. XRPD indexing is the process of determining the size, shape, and symmetry of the crystallographic unit cells of the crystalline components responsible for a set of peaks in an XRPD pattern. Crystalline Compound A-MsA Form 1 was sampled with Cu-Kα radiation and the indexing results are summarized in Table 7 below.

[0232] [Table 10]

[0233] Thermal analysis: DSC and TGA of crystalline compound A-MsA Form 1 are shown in Figure 11. Typical DSC of crystalline compound A-MsA Form 1 showed an onset of melting at 250°C. TGA of crystalline compound A-MsA Form 1 showed a weight loss of 0.2% before decomposition.

[0234] Hygroscopicity Analysis: The hygroscopicity profile of crystalline compound A-MsA Form 1 is shown in Figure 12. A typical DVS of crystalline compound A-MsA Form 1 showed a weight gain of about 1.2% at 95% RH.

[0235] Example 5: Crystalline Compound A-MsA Form 2 Crystalline Compound A-MsA Form 2 was prepared by slurrying 1 equivalent of MsA and Compound A in 90 / 10 THF / water (v / v) solvent at ambient conditions.

[0236] Powder X-ray diffraction: The XRPD pattern of the crystalline Compound A-MsA Form 2 material is shown in FIG.

[0237] Thermal analysis: The DSC of crystalline compound A-MsA form 2 is shown in Figure 15. The typical DSC of crystalline compound A-MsA form 2 showed an onset of melting at 38.0° C. and an onset of endothermic event at 177.1° C. The TGA of crystalline compound A-MsA form 2 showed a weight loss of about 0.3% before decomposition (see Figure 16).

[0238] Example 6: Crystalline Compound A-TsA Form 1 and Form 5 Crystalline Compound A-TsA Form 1 was prepared by slurrying Compound A with one molar equivalent of p-toluenesulfonic acid in acetonitrile at ambient conditions.

[0239] Powder X-ray diffraction: The XRPD pattern of the crystalline Compound A-TsA Form 1 material is shown in FIG.

[0240] Variable temperature X-ray diffraction (VTXRD) of crystalline compound A-TsA Form 1 showed recrystallization at temperatures >180° C., and the new crystalline form was designated crystalline compound A-TsA Form 5. The VTXRD pattern is shown in FIG.

[0241] Thermal Analysis: The DSC and TGA patterns of crystalline compound A-TsA Form 1 are shown in Figure 19. Typical DSC of crystalline compound A-TsA Form 1 showed endothermic onsets at 193.9°C and 258.4°C. TGA of crystalline compound A-TsA Form 1 showed a weight loss of about 0.07% before decomposition.

[0242] Example 7: Crystalline Compound A-TsA Form 3 Crystalline Compound A-TsA Form 3 was prepared by slurrying 1 molar equivalent of p-toluenesulfonic acid and Compound A in 90 / 10 EtOH / water (v / v) at ambient conditions.

[0243] Powder X-ray diffraction: The XRPD pattern of crystalline compound A-TsA Form 3 is shown in FIG.

[0244] Thermal Analysis: The DSC and TGA patterns of crystalline compound A-TsA Form 3 are shown in Figure 23. Typical DSC of crystalline compound A-TsA Form 3 showed endothermic onsets at 161.0°C and 248.9°C. TGA of crystalline compound A-TsA Form 3 showed a weight loss of about 0.48% before decomposition.

[0245] Example 8: Crystalline Compound A-TsA Form 4 Crystalline Compound A-TsA Form 4 was prepared by slurrying one molar equivalent of p-toluenesulfonic acid and Compound A in EtOH at ambient conditions. Alternatively, this compound was produced by drying Compound A-TsA Form 1 under vacuum at a temperature of 95° C. to 103° C. for 1 day, then at a temperature of 107° C. to 109° C. for 3 days, or at a temperature of 150° C. to 170° C. for 1 day.

[0246] Scale-up of Compound A-TsA salt Form 4 was prepared by desolvation of Compound A-isopropanol solvate of TSA salt Form 1. The procedure consisted of stirring 3.5 g of Compound A and 1 molar equivalent of p-toluenesulfonic acid (1.08 g) in isopropanol (60 ml) at 60° C. in a Mettler Toledo EasyMax controlled laboratory reactor equipped with an overhead stirrer. The slurry was stirred at 60° C. for 1 day and then cooled to 20° C. at 0.1° C. / min. The solid was isolated by vacuum filtration and washed twice with isopropanol (10 ml). XRPD analysis showed that the material was a mixture of tosylate salt Form 1 and a small amount of the free form of the isopropanol solvate. To drive the reaction to completion, the solid was reslurried in isopropanol (30 ml) with approximately 0.15 molar equivalent of p-toluenesulfonic acid (0.21 g) at ambient temperature for 4 days. The solid was isolated by vacuum filtration and washed twice with isopropanol (10 ml). XRPD analysis indicated that the solid consisted of tosylate salt Form 1, but still contained traces of the free form isopropanol solvate. The solid was reslurried with 0.25 molar equivalents of p-toluenesulfonic acid (0.34 g) in isopropanol (50 ml) at 60° C. After stirring for 1 day, the solid was isolated by vacuum filtration. The wet cake was washed with isopropanol (15 ml) and analyzed by XRPD. The XRPD pattern was consistent with Compound A-TsA salt Form 1 and a small amount of Compound A-TsA salt Form 4. This material was dried under vacuum at 145° C., resulting in complete conversion to Compound A-TsA salt Form 4 by XRPD. The XRPD pattern of the crystalline Compound A-TsA salt Form 4 material is shown in FIG. 24a, and the XRPD peaks are shown in Table 7.

[0247] [Table 11-1] [Table 11-2]

[0248] Single crystal data: Table 8 summarizes the crystallographic data of crystalline compound A-TsA Form 4. The molecular structure of crystalline compound A-TsA Form 4 as found from X-ray crystal structure determination is shown in Figure 24b.

[0249] [Table 12]

[0250] Thermal Analysis: DSC and TGA patterns of crystalline compound A-TsA Form 4 are shown in Figure 25. Typical DSC of crystalline compound A-TsA Form 4 showed melting onset at 253°C. TGA of crystalline compound A-TsA Form 4 showed a weight loss of 0.145% before decomposition.

[0251] Solid-state NMR: Solid state of crystalline compound A-TsA form 4 19 The F NMR spectrum is shown in Figure 26. Two peaks are observed at -96.93 and -101.60 ppm.

[0252] Example 9: Crystalline Compound A-TsA Form 5 Crystalline Compound A-TsA Form 5 was prepared by heating Crystalline Compound A-TsA Form 1 above 180°C.

[0253] Powder X-ray diffraction: The XRPD pattern is shown in FIG.

[0254] Example 10: Crystalline Compound A-DiTsA Form 6 Crystalline Compound A-DiTsA Form 6 was prepared by slurrying Compound A with 2 molar equivalents of p-toluenesulfonic acid in acetonitrile at high throughput settings. Attempts to scale up this compound were unsuccessful.

[0255] Powder X-ray diffraction: The XRPD pattern is shown in FIG.

[0256] Example 11: Crystalline Compound A - Sulfate Form 1 Crystalline Compound A-Sulfate Salt Form 1 was prepared by slurrying Compound A with 1 equivalent of sulfuric acid in acetonitrile at ambient conditions.

[0257] Powder X-ray diffraction: The XRPD pattern is shown in FIG.

[0258] Thermal Analysis: DSC and TGA of crystalline Compound A-sulfate salt Form 1 are shown in Figure 31. Typical DSC of crystalline Compound A-sulfate salt Form 1 showed endothermic onsets at 182.3°C and 263.7°C. TGA of crystalline Compound A-sulfate salt Form 1 showed a weight loss of 6.47% before decomposition.

[0259] Hygroscopicity Analysis: The hygroscopicity profile of crystalline Compound A-sulfate salt Form 1 is shown in Figure 32. Dynamic Vapor Sorption (DVS) of crystalline Compound A-sulfate salt Form 1 suggests that the sulfate salt deliquesces at 90% RH.

[0260] Example 12: Amorphous Compound A Amorphous Compound A was prepared by dissolving 1.99 g of Compound A-variable hydrate form 2 (see Example 13) in 100 mL of acetone and shaking to form a yellow solution. This solution was then spray dried at a spray rate of 2 mL / min under the following conditions: inlet temperature 54° C., outlet temperature 54° C., aspirator 95%, drying air flow rate 0.55 kg / min, nozzle air 6.0 sL / min, nozzle cooling 20° C. The amorphous product was collected and dried in a vacuum oven at 40° C. and −10 bar pressure for 2.5 hours to remove residual acetone.

[0261] Powder X-ray Diffraction: The XRPD pattern of amorphous Compound A is shown in FIG.

[0262] Thermal Analysis: The DSC of amorphous Compound A is shown in Figure 38. A typical DSC of amorphous Compound A shows a glass transition temperature (T g) was shown. The TGA-IR of amorphous Compound A is shown in Figure 34. As shown in Figure 35, the TGA-IR of amorphous Compound A showed a weight loss of 1.05% of water molecules at a temperature lower than 100°C.

[0263] Example 13: Compound A - Variable Hydrate Form 2 Compound A-variable hydrate Form 2 was prepared by slurrying a mixture of Compound A-methanol Form 1 and Compound A-ethanol Form 1 products in water for 24 hours. The product was then filtered and air-dried.

[0264] Alternatively, Compound A-variable hydrate form 2 was prepared by mixing Compound A in a mixed solvent of methanol and ethanol. Compound A first formed a Compound A-methanol and Compound A-ethanol solvate mixture, which was then slurried in water to initiate conversion to the Compound A-variable hydrate form 2 product. To achieve complete conversion, the Compound A-variable hydrate form 2 product was filtered and dried at elevated temperature (e.g., 50° C.) overnight to remove any remaining organic solvent.

[0265] X-Ray Powder Diffraction: The XRPD pattern of Compound A-variable hydrate form 2 is shown in FIG. 36 and the XRPD peaks are listed in Table 9.

[0266] [Table 13-1] [Table 13-2]

[0267] Thermal analysis: DSC of Compound A-variable hydrate form 2 is shown in Figure 37. Typical DSC of Compound A-variable hydrate form 2 showed a dehydration onset of 51°C and a melting point of 136°C. TGA of Compound A-variable hydrate form 2 is shown in 38. TGA of Compound A-variable hydrate form 2 showed a weight loss of 2.0% of water molecules at temperatures below 100°C.

[0268] Hygroscopicity Analysis: The hygroscopicity profile of Compound A-variable hydrate form 2 is shown in Figure 39. Dynamic Vapor Sorption (DVS) of Compound A-variable hydrate form 2 showed a weight gain of about 3.4% at 95% RH.

[0269] Example 14: Anhydrous Compound A Form 3 Anhydrous Compound A Form 3 was obtained by heating Compound A-THF solvate to 150° C. and holding for 3 minutes, followed by equilibration at room temperature.

[0270] X-ray Powder Diffraction: The XRPD pattern of anhydrous Compound A Form 3 is shown in FIG.

[0271] Thermal Analysis: The DSC of anhydrous Compound A Form 3 is shown in Figure 41. A typical DSC of anhydrous Compound A Form 3 showed an onset of melting at 196.5°C.

[0272] Hygroscopicity Analysis: The hygroscopicity profile of anhydrous Compound A Form 3 is shown in Figure 42. Dynamic Vapor Sorption (DVS) of anhydrous Compound A Form 3 showed a weight gain of about 1.5% at 95% RH.

[0273] Example 15: Anhydrous Compound A Form 4 Anhydrous Compound A Form 4 was obtained by slurrying anhydrous Compound A Form 3 and Compound A-variable hydrate Form 2 (Example 13) in heptane at 40° C. for 5 days.

[0274] X-ray Powder Diffraction: The XRPD pattern of anhydrous Compound A Form 4 is shown in FIG.

[0275] Example 16: Anhydrous Compound A Form 5 Anhydrous Compound A Form 5 was obtained by slurrying 350 mg of a mixture of anhydrous Compound A Form 3 and Compound A-variable hydrate Form 2 (Example 13) in 18 mL of heptane at a temperature of 70° C. for 1 day. The solid was then removed from the hot plate, filtered, washed with 5 mL of heptane; it was then dried overnight under nitrogen blowing.

[0276] X-ray Powder Diffraction: The XRPD pattern of anhydrous Compound A Form 5 is shown in FIG.

[0277] Thermal Analysis: The DSC and TGA patterns of anhydrous Compound A Form 5 are shown in Figure 45. Typical DSC of anhydrous Compound A Form 5 showed an onset of melting at 136.5° C. TGA of anhydrous Compound A Form 5 showed a weight loss of 0.17%.

[0278] Hygroscopicity Analysis: The hygroscopicity profile of anhydrous Compound A Form 5 is shown in Figure 46. Dynamic Vapor Sorption (DVS) of anhydrous Compound A Form 5 showed that the compound rehydrated to Compound A-variable hydrate Form 2 (Example 13).

[0279] Example 17: Anhydrous Compound A Form 6 Anhydrous Compound A Form 6 was obtained by slurrying a mixture of anhydrous Compound A Form 3 and Compound A-variable hydrate Form 2 (Example 13) in heptane at a temperature of 80° C. overnight.

[0280] X-ray Powder Diffraction: The XRPD pattern of anhydrous Compound A Form 6 is shown in FIG.

[0281] Thermal Analysis: DSC and TGA of anhydrous Compound A Form 6 are shown in Figure 48. Typical DSC of anhydrous Compound A Form 6 showed an onset of melting at 186.4° C. TGA of anhydrous Compound A Form 6 showed a weight loss of 0.38%.

[0282] Example 18: Anhydrous Compound A Form 7 Anhydrous Compound A Form 7 was obtained by slurrying anhydrous Compound A Form 3 and Compound A-variable hydrate Form 2 (Example 13) in heptane at a temperature of 70° C. for 3 days.

[0283] X-ray Powder Diffraction: The XRPD pattern of anhydrous Compound A Form 7 is shown in FIG.

[0284] Example 19: Anhydrous Compound A Form 8 Anhydrous Compound A Form 8 was obtained by slurrying anhydrous Compound A Form 3 and Compound A-variable hydrate Form 2 (Example 13) in toluene at a temperature of 50° C. for 3 days.

[0285] X-ray Powder Diffraction: The XRPD pattern of anhydrous Compound A Form 8 is shown in FIG.

[0286] Thermal Analysis: DSC and TGA of anhydrous Compound A Form 8 are shown in Figure 51. Typical DSC of anhydrous Compound A Form 8 showed melting onset at 156.3° C. and 185.9° C. TGA of anhydrous Compound A Form 8 showed a weight loss of 0.73%.

[0287] Example 20: Crystalline Compound A Form 1 Compound A was purified by silica gel column chromatography on a pre-packed Redi Sep column (12 g) and combi-flash using 20%-100% EtOH in hexane as eluent to obtain crystalline Compound A Form 1. The fractions containing the desired product were then concentrated under reduced pressure and the residue was dissolved in acetonitrile / water solvent mixture and lyophilized.

[0288] X-ray Powder Diffraction: The XRPD pattern of crystalline Compound A Form 1 is shown in FIG.

[0289] Example 21: Crystalline Compound A-THF solvate Crystalline Compound A-THF solvates were prepared by slurrying Compound A in various solvents: a) 50 mg / mL THF solution, b) 50-50 THF / water mixed solvent, c) 50-50 THF / methanol mixed solvent, or d) 50-25-25 THF-NMP-water mixed solvent.

[0290] Powder X-ray Diffraction: The XRPD pattern of crystalline Compound A-THF solvate is shown in FIG.

[0291] Thermal Analysis: The DSC and TGA of crystalline Compound A-THF solvate are shown in Figure 54. The typical DSC of crystalline Compound A-THF solvate showed an onset of melting at 122.6°C and an onset of desolvation at 191.5°C. The TGA of crystalline Compound A-THF solvate showed a weight loss of 11.4%, corresponding to the desolvation endotherm of one molar equivalent of THF molecules.

[0292] Single Crystal Data: Table 10 summarizes the crystallographic data for crystalline Compound A-THF solvate.

[0293] [Table 14]

[0294] Example 22: Crystalline Compound A - Ethanol Solvate Crystalline Compound A-ethanol solvate was prepared by slurrying Compound A in ethanol.

[0295] Powder X-ray Diffraction: The XRPD pattern of crystalline Compound A-ethanol solvate is shown in FIG.

[0296] Thermal Analysis: The TGA of crystalline Compound A-ethanol solvate is shown in Figure 56. The TGA of crystalline Compound A-ethanol solvate showed a weight loss of 7.58%, corresponding to the loss of one molar equivalent of ethanol molecule. The DSC of crystalline Compound A-ethanol solvate is shown in Figure 57. The typical DSC of crystalline Compound A-ethanol solvate showed endothermic onsets at 131.8°C, 165.6°C, and 198.1°C.

[0297] Example 23: Crystalline Compound A - Propanol Solvate Crystalline Compound A-propanol solvate was prepared by slurrying Compound A in 1-propanol.

[0298] X-Ray Powder Diffraction: The XRPD pattern of crystalline Compound A-propanol solvate is shown in FIG.

[0299] Thermal Analysis: The TGA and DSC of crystalline Compound A-propanol solvate are shown in Figure 59. The TGA of crystalline Compound A-propanol solvate showed a weight loss of 9.95%, corresponding to the loss of one molar equivalent of 1-propanol molecules. The typical DSC of crystalline Compound A-propanol solvate showed an onset of melting at 112.2°C and 194.2°C.

[0300] Example 24: Crystalline Compound A - Isopropyl Alcohol (IPA) Solvate Crystalline Compound A-IPA solvate was prepared by slurrying Compound A in a 50-50 1-propanol / water mixture.

[0301] Powder X-ray Diffraction: The XRPD pattern of the crystalline Compound A-IPA solvate is shown in FIG.

[0302] Thermal Analysis: The TGA and DSC of the crystalline Compound A-IPA solvate are shown in Figure 61. The TGA of the crystalline Compound A-IPA solvate showed a weight loss of 8.5%, corresponding to the loss of one molar equivalent of isopropyl alcohol molecules. The typical DSC of the crystalline Compound A-IPA solvate showed endothermic onsets at 114.6°C; 158.7°C; and 194.9°C.

[0303] Example 25: Crystalline Compound A - Methanol Solvate Crystalline Compound A-methanol solvate was prepared by slurrying Compound A in methanol.

[0304] Powder X-ray Diffraction: The XRPD pattern of crystalline Compound A-methanol solvate is shown in FIG.

[0305] Example 26: Crystalline Compound A - Isopropyl Acetate (IPAc) Solvate Crystalline Compound A-IPAc solvate was prepared by slurrying Compound A in isopropyl acetate.

[0306] Powder X-ray Diffraction: The XRPD pattern of the crystalline Compound A-IPAc solvate is shown in FIG.

[0307] Example 27: Crystalline Compound A - Acetone Solvate Crystalline Compound A-acetone solvate was prepared by slurrying Compound A in acetone.

[0308] Powder X-ray Diffraction: The XRPD pattern of crystalline Compound A-acetone solvate is shown in FIG.

[0309] Example 28: Crystalline Compound A - Cyclopentyl Methyl Ether (CPME) Solvate Crystalline Compound A-CPME solvate was prepared by slurrying Compound A in cyclopentyl methyl ether.

[0310] Powder X-ray Diffraction: The XRPD pattern of crystalline Compound A-CPME solvate is shown in FIG.

[0311] Example 29: Crystalline Compound A - Dioxane Solvate Crystalline Compound A-dioxane solvate was prepared by slurrying Compound A in dioxane.

[0312] Powder X-ray Diffraction: The XRPD pattern of crystalline Compound A-dioxane solvate is shown in FIG.

[0313] Example 30: Crystalline Compound A - Ethyl Acetate (EtOAc) Solvate Crystalline Compound A-EtOAc solvate was prepared by slurrying Compound A in ethyl acetate.

[0314] Powder X-ray Diffraction: The XRPD pattern of crystalline Compound A-EtOAc solvate is shown in FIG.

[0315] Example 31: Crystalline Compound A - Acetonitrile (MeCN) Solvate Crystalline Compound A-MeCN solvate was prepared by slurrying Compound A in acetonitrile.

[0316] Powder X-ray Diffraction: The XRPD pattern of the crystalline Compound A-MeCN solvate is shown in FIG.

[0317] Example 32: Crystalline Compound A - Methyl tert-butyl ether (MTBE) solvate Crystalline Compound A-MTBE solvate was prepared by slurrying Compound A in methyl tert-butyl ether.

[0318] Powder X-ray Diffraction: The XRPD pattern of crystalline Compound A-CMTBE solvate is shown in FIG.

[0319] Example 33: Crystalline Compound A - Toluene Solvate Crystalline Compound A-toluene solvate was prepared by slurrying Compound A in toluene at 25° C. for 18 hours.

[0320] Powder X-ray Diffraction: The XRPD pattern of crystalline Compound A-toluene solvate is shown in FIG.

[0321] Example 34: Crystalline Compound A - Dodecyl Sulfate Crystalline Compound A-dodecyl sulfate was prepared by slurrying 100 mg of Compound A-HCl in 0.5% sodium dodecyl sulfate (SDS) with or without 0.01 N HCl for 3 hours at 37°C. The solid was then removed, filtered, washed with 1 mL of deionized water, and dried overnight under nitrogen sparging. A new crystalline form was obtained, and solution NMR analysis showed a 1:1 ratio of API:dodecyl sulfate, with assay confirming a Compound A content of 69%, which correlated with 1 equivalent of dodecyl sulfate.

[0322] Powder X-ray Diffraction: The XRPD pattern of crystalline Compound A-dodecyl sulfate is shown in FIG.

[0323] Thermal Analysis: The TGA and DSC of crystalline Compound A-dodecyl sulfate are shown in Figure 72. The TGA of crystalline Compound A-dodecyl sulfate showed a weight loss of 21.1%. The typical DSC of crystalline Compound A-dodecyl sulfate showed an onset melting temperature of 75.8°C and decomposition at 174.8°C.

[0324] Example 35: Crystalline Compound A - Dimethylformamide (DMF) Solvate Hydrate Compound A-HCl Form 1 was dissolved in DMF solvent to prepare crystalline Compound A-DMF solvate hydrate. The solution was then filtered to remove any solid particles remaining in the solution. The clear solution was allowed to slowly evaporate the solvent at room temperature in a fume hood. Single crystals were observed after one week.

[0325] X-Ray Powder Diffraction: The XRPD pattern of crystalline Compound A-DMF solvate hydrate is shown in FIG.

[0326] Thermal Analysis: The DSC of crystalline Compound A-DMF solvate hydrate is shown in Figure 74. A typical DSC of crystalline Compound A-DMF solvate hydrate showed an onset of melting at 107.8°C.

[0327] Single Crystal Data: In the crystal structure of the provided crystal, the DMF molecule was shown to be disordered, with the water molecule improving the fractional occupancy to 0.25. The DMF molecule was shown not to be hydrogen bonded to Compound A. Table 11 summarizes the crystallographic data for the crystalline Compound A-DMF solvate hydrate.

[0328] [Table 15]

[0329] Example 36: Crystalline Compound A - Dimethylacetamide (DMAC) Solvate Compound A-HCl Form 1 was dissolved in DMAC solvent to prepare crystalline Compound A-DMAC solvate. The solution was then filtered to remove any solid particles remaining in the solution. The clear solution was allowed to slowly evaporate the solvent at room temperature in a fume hood. Single crystals were observed after one week.

[0330] Powder X-ray diffraction: The XRPD pattern of the crystalline compound A-DMAC solvate is shown in FIG.

[0331] Thermal Analysis: The DSC of crystalline Compound A-DMAC solvate is shown in Figure 76. A typical DSC of crystalline Compound A-DMAC solvate showed an onset of melting at about 150°C.

[0332] Single Crystal Data: In the crystal structure of the provided crystals, the DMAC molecule was shown to be disordered. However, the DMAC molecule was still shown to be hydrogen bonded to Compound A. Table 12 summarizes the crystallographic data for the crystalline Compound A-DMAC solvate.

[0333] [Table 16]

[0334] Example 37: Crystalline Compound A - Monobesylate Hydrate Form 1 Crystalline Compound A-monobesylate hydrate Form 1 was prepared by dissolving 92.6 mg of Compound A and 29.3 mg of benzenesulfonic acid in 1 mL of methanol solvent. The solution was then stirred at 60° C. for 1 day. A slurry resulted and the solid was isolated by vacuum filtration. The solid was air-dried for 1 hour and then analyzed.

[0335] Powder X-Ray Diffraction: The XRPD pattern of crystalline Compound A-monobesylate hydrate Form 1 is shown in FIG.

[0336] Thermal Analysis: The DSC and TGA of crystalline Compound A-monobesylate hydrate Form 1 are shown in Figure 78. The onset melting temperature was about 230.8°C. The TGA of crystalline Compound A-monobesylate hydrate Form 1 showed a weight loss of about 1.3% up to 142.3°C.

[0337] Example 38: Crystalline Compound A-Caffeine Cocrystal Form 1 Compound A-caffeine cocrystal Form 1 was prepared by slow cooling from 70° C. to 5° C. in acetonitrile using a 1:1 molar ratio of Compound A:caffeine. The resulting product contained the remaining Compound A starting material mixed with caffeine cocrystal Form 1, as well as other impurities that were not further identified. The resulting product was then further purified by heating the mixture to 167° C. in a DSC furnace under a nitrogen stream to form pure Compound A-caffeine cocrystal Form 1.

[0338] Powder X-ray Diffraction: The XRPD pattern of crystalline Compound A-caffeine co-crystal Form 1 is shown in Figure 79. The single crystal structure data for crystalline Compound A-caffeine co-crystal Form 1 is shown in Table 13 below.

[0339] [Table 17]

[0340] Thermal Analysis: The DSC and TGA patterns of crystalline Compound A-caffeine cocrystal Form 1 are shown in Figure 80. DSC showed an onset of melting at about 169.5°C. TGA of crystalline Compound A-caffeine cocrystal Form 1 showed a weight loss of about 0.39% to 135.3°C.

[0341] Hygroscopicity Analysis: The hygroscopicity profile of crystalline Compound A-caffeine cocrystal Form 1 is shown in Figure 81. Dynamic Vapor Sorption (DVS) of crystalline Compound A-caffeine cocrystal Form 1 showed a weight gain of less than 0.20% at about 95% RH.

[0342] Example 39: Crystalline Compound A - Citric Acid Co-Crystal Form 1 In an experiment using a 1:1 molar ratio of compound A:citric acid in ethyl acetate, and slowly cooling from 70°C to 5°C, crystalline compound A-citric acid co-crystal 1 was obtained.

[0343] Powder X-ray Diffraction: The XRPD pattern of crystalline Compound A-citric acid co-crystal Form 1 is shown in Figure 82. Crystalline Compound A-citric acid co-crystal Form 1 was collected with Cu-Kα radiation and the indexing results are shown in Table 14 below.

[0344] [Table 18]

[0345] Thermal Analysis: The DSC and TGA of crystalline Compound A-citric acid co-crystal Form 1 are shown in Figure 83. The onset melting temperature was shown to be about 107.7°C. The TGA of crystalline Compound A-citric acid co-crystal Form 1 showed a weight loss of about 6.3% by 140.2°C, 0.8 mg.

[0346] Example 40: Crystalline Compound A - Citric Acid Co-Crystal Form 2 Crystalline Compound A-citric acid co-crystal 2 was obtained by slow cooling experiments in acetonitrile from 70 °C to refrigerator temperature using a 1:2 molar ratio of Compound A to citric acid. The sample was first oiled out and stirred at 5 °C for 3 days to produce an off-white precipitate.

[0347] Powder X-ray Diffraction: The XRPD pattern of crystalline Compound A-citric acid co-crystal Form 2 is shown in Figure 84. Crystalline Compound A-citric acid co-crystal Form 2 was collected with Cu-Kα radiation and the indexing results are shown in Table 15 below.

[0348] [Table 19]

[0349] Thermal Analysis: The DSC and TGA patterns of crystalline Compound A-citric acid co-crystal Form 2 are shown in Figure 85. DSC showed an endotherm onset at about 93.8°C. TGA of crystalline Compound A-citric acid co-crystal Form 2 showed a weight loss of about 5.3% to 135.3°C, 0.6 mg.

[0350] Example 41: Crystalline Compound A - Saccharin - Co-Crystal Form 1 Crystalline Compound A-saccharin cocrystal Form 1 was prepared experimentally using a 1:1 molar ratio of Compound A to saccharin in acetonitrile by slow cooling from 70° C. to 5° C.

[0351] Powder X-ray Diffraction: The XRPD pattern of crystalline Compound A-saccharin co-crystal Form 1 is shown in Figure 86. Crystalline Compound A-saccharin co-crystal Form 1 was collected with Cu-Kα radiation and the indexing results are shown in Table 16 below.

[0352] [Table 20]

[0353] Thermal Analysis: The DSC and TGA of crystalline Compound A-saccharin cocrystal Form 1 are shown in Figure 87. The DSC showed an onset of melting at about 177.0°C. The TGA of crystalline Compound A-saccharin cocrystal Form 1 showed a weight loss of 0.3 mg, about 2.2%, up to 100.2°C.

[0354] Hygroscopicity Data: The hygroscopicity profile of crystalline Compound A-saccharin cocrystal Form 1 is shown in Figure 88. Dynamic Vapor Sorption (DVS) of crystalline Compound A-saccharin cocrystal Form 1 showed a weight gain of approximately 0.3% at 95% RH.

[0355] Example 42: Crystalline Compound AL-Tartaric Acid Co-Crystal Form 1 Crystalline Compound A L-Tartaric Acid Co-Crystal Form 1 was prepared by experimental slow cooling from 70° C. to 5° C. in acetonitrile using a 1:1 molar ratio of Compound A and L-tartaric acid.

[0356] Powder X-ray diffraction: The XRPD pattern of the crystalline compound AL-tartaric acid co-crystal Form 1 is shown in Figure 89. The crystalline compound AL-tartaric acid co-crystal Form 1 was collected with Cu-Kα radiation and the indexing results are shown in Table 17 below.

[0357] [Table 21]

[0358] Thermal Analysis: DSC and TGA of crystalline Compound A L-Tartaric acid co-crystal Form 1 are shown in Figure 90. DSC showed an onset of melting at about 157.0°C. TGA of crystalline Compound A-Tartaric acid co-crystal Form 1 showed a weight loss of 0.2 mg, about 2.5%, to 140.2°C.

[0359] Hygroscopicity Data: The hygroscopicity profile of crystalline compound AL-tartaric acid co-crystal Form 1 is shown in Figure 91. Dynamic vapor sorption of crystalline compound AL-tartaric acid co-crystal Form 1 showed a weight gain of about 4.75% at 95% RH.

[0360] Example 43: Crystalline Compound A - Urea Co-Crystal Form 1 Crystalline Compound A-urea co-crystal Form 1 was prepared experimentally in acetonitrile using a 2:1 molar ratio of Compound A to urea with slow cooling from 70° C. to freezer temperatures of −15° C. to −25° C.

[0361] Powder X-ray Diffraction: The XRPD pattern of crystalline Compound A-urea co-crystal Form 1 is shown in Figure 92. Crystalline Compound A-urea co-crystal Form 1 was collected with Cu-Kα radiation and the indexing results are shown in Table 18 below.

[0362] [Table 22]

[0363] Thermal Analysis: The DSC and TGA of crystalline Compound A-urea co-crystal Form 1 are shown in Figure 93. The DSC showed a first endotherm onset at about 106.4°C and a second endotherm onset at about 156.8°C. The TGA of crystalline Compound A-urea co-crystal Form 1 showed a weight loss of 0.5 mg, about 4.5%, to 155.2°C.

[0364] Hygroscopicity Data: The hygroscopicity profile of crystalline Compound A-urea cocrystal Form 1 is shown in Figure 94. Dynamic Vapor Sorption (DVS) of crystalline Compound A-urea cocrystal Form 1 showed less than 40% weight gain at 95% RH.

[0365] Solubility, Powder Dissolution (PD) and Intrinsic Dissolution Rate (IDR) Testing Example 44: PD and IDR Study of Compound A-HCl Form 1 Compared to Various Forms of Non-Salt Compound A The solubility of various forms of Compound A and Compound A-HCl Form 1 was measured in fasted-state simulated gastric fluid (FaSSGF), fasted-state simulated intestinal fluid (FaSSIF), fed-state simulated intestinal fluid (FeSSIF) and water. The results of the powder dissolution measurement study showed that crystalline Compound A-HCl Form 1 was more soluble than Compound A-variably hydrated Form 2 or Compound A-anhydrous Form 3, but was less soluble than amorphous Compound A. The solubility and IDR data are shown in Tables 19 and 20, respectively. The data show that crystalline Compound A-HCl Form 1 has superior solubility and IDR to all forms tested here.

[0366] [Table 23]

[0367] [Table 24]

[0368] Example 45: Biological Data Dog crossover PK study of Compound A-HCl Form 1, Compound A-anhydrous Form 3 and amorphous Compound A. A total of three male dogs were initially assigned to the study. All animals were fasted for at least 8 hours prior to dosing and were fasted until the first 4 hours of blood sampling (where applicable, food was returned within 30 minutes after the last blood sample was taken at the 4-hour sampling interval).

[0369] Each animal received an oral gavage dose (PO) of the appropriate test article solution containing Compound A as outlined in the study design table below. Oral gavage dosing solutions were constantly stirred during dosing. The gavage tube was washed with approximately 10 mL of tap water after dosing (before removing the gavage tube). A minimum of 10 days washout period was allowed between dosing of each phase.

[0370] [Table 25]

[0371] The results of the dog PK crossover study are shown in Table 22. As shown in Figure 95, Compound A-HCl Form 1 showed lower exposure than the amorphous Compound A form. However, Compound A-HCl Form 1 showed approximately twice the exposure compared to Compound A-Anhydrous Form 3, suggesting higher solubility than Compound A-Anhydrous Form 3.

[0372] [Table 26]

[0373] Example 46: PD and IDR Study of Compound A-HCl Form 1 Compared to Compound A-MsA and A1-TsA The solubilities of Compound A-HCl Form 1, Compound A-MsA Form 1 and Compound A-TsA Form 4 were measured in fed-state simulated intestinal fluid (FeSSIF) at pH 6.5.

[0374] All three salts showed higher kinetic solubility and faster dissolution rates than Compound A in FaSSIF. The dissolution rate of the tosylate (A-TsA or A-TSA) salt form 4 is better than that of the mesylate (A-MsA or A-MSA) salt form 1, which is better than that of the hydrochloride salt form 1. All three salts can be converted to free base, but maintain a supersaturated state in FaSSIF for some time, indicating that they can be expected to be well absorbed in vivo when used in pharmaceutical dosage forms. The data of the solubility test results are shown in Table 23.

[0375] [Table 27]

[0376] The foregoing description has been given for clarity of understanding only, and no unnecessary limitations should be understood therefrom, since modifications within the scope of the invention will be apparent to those skilled in the art.

[0377] Throughout this specification and the claims that follow, unless the context clearly indicates otherwise, the word "comprise" and variations such as "comprises" and "comprising" are understood to imply the inclusion of a stated integer or step, or group of integers or steps, but not the exclusion of any other integers or steps, or group of integers or steps.

[0378] Throughout this specification, when a composition is described as comprising a component or material, it is contemplated that the composition may also consist essentially of or consist of any combination of the described components or materials, unless otherwise stated. Similarly, when a method is described, it is contemplated that the method may also consist essentially of or consist of any combination of the described steps, unless otherwise stated. The invention illustratively disclosed herein may suitably be practiced in the absence of any element or step not specifically disclosed herein.

[0379] As will be apparent to those skilled in the art upon reading this disclosure, each of the individual embodiments described and illustrated herein has distinct components and features which may be readily separated from or combined with the features of any other several embodiments without departing from the scope or spirit of the disclosure. Any described method may be carried out in the order of events described or in any other order which is logically possible.

[0380] The implementation of the methods disclosed herein and their individual steps can be performed manually and / or with the aid of electronic devices or automation provided by electronic devices. Although the processes have been described with reference to specific embodiments, one of ordinary skill in the art will readily appreciate that other ways of performing the acts associated with the methods may be used. For example, unless otherwise stated, the order of various steps may be changed without departing from the scope or spirit of the methods. Furthermore, some of the individual steps may be combined, omitted, or further subdivided into additional steps.

[0381] The use of the terms "a," "an," and "the" and similar referents in the context of this disclosure (particularly in the context of the claims) are intended to encompass both the singular and the plural unless otherwise indicated. The recitation of ranges of values ​​herein is merely intended to serve as a shorthand for referring to each separate value and each end point within the range individually, unless otherwise specified herein, and each separate value and end point is incorporated herein as if individually set forth herein. The use of any and all examples or representative language (e.g., "etc.") provided herein is intended to further clarify the disclosure and does not impose limitations on the scope of the disclosure unless otherwise indicated. No language in this specification should be construed as indicating any non-claimed element as essential to the practice of the disclosure.

[0382] All patents, publications, and references cited herein are hereby incorporated by reference in their entirety. In the event of any conflict between this disclosure and the incorporated patents, publications, and references, this disclosure shall control.

Claims

1. A salt, hydrate, solvate or co-crystal of compound A having the following chemical structure: 【Chemical 1】 Or a solid form of compound A including the crystalline anhydrous form, its salt, solvate, or co-crystal.

2. Hydrochloride (compound A-HCl), mesylate (compound A-MsA), tosylate (compound A-TsA), sulfate (compound A-sulfate), variable hydrate (compound A-variable hydrate), tetrahydrofuran solvate (compound A-THF), ethanol solvate (compound A-ethanol), 1-propanol solvate (compound A-1-propanol), isopropyl alcohol solvate (compound A-IPA), methanol solvate (compound A-methanol), isopropyl acetate solvate (compound A-IPAc), acetone solvate (compound A-acetone), cyclopentyl methyl ether solvate (compound A-CPME), dioxane solvate (compound A-dioxane), ethyl acetate solvate (compound A-EtOAc), acetonitrile solvate (compound A-MeCN), methyl tert-butyl ether solvate (compound A-MTBE), toluene solvate (compound A-toluene), dodecyl sulfate (compound A-dodecyl sulfate), dimethylformamide (DMF) solvate hydrate (compound A-DMF-hydrate), dimethylacetamide (DMAC) solvate (compound A-DMAC), monobenzylate hydrate (compound A-benzylate-hydrate), caffeine co-crystal (compound A-caffeine), citric acid co-crystal (compound A-citric acid), saccharin co-crystal (compound A-saccharin), L-tartaric acid co-crystal (compound A-L-tartaric acid), or urea co-crystal (compound A-urea); or a salt, hydrate, solvate or co-crystal according to claim 1 selected from their solid forms.

3. The solid form of compound A-HCl according to claim 2.

4. It is crystalline form 1 and in the solid state １９ The solid form of compound A-HCl according to claim 3, characterized in that the peaks of 19F NMR are at -91 and -103 ± 0.5 ppm.

5. The crystalline form 1 of compound A-HCl according to claim 4, further characterized in that the peaks of the XRPD pattern using CuKα radiation are at 7.5, 16.9, and 20.2 ± 0.2° 2θ.

6. The crystalline form 1 of compound A-HCl according to claim 5, further characterized in that the peaks of the XRPD pattern using CuKα radiation are at 12.8, 18.2, 22.7, 23.6, 24.8, and 26.1 ± 0.2° 2θ.

7. The crystalline form 1 of compound A-HCl according to claim 6, further characterized in that the peaks of the XRPD pattern using CuKα radiation are at 10.9, 14.5, 15.7, 15.9, 19.8, 20.6, 21.6, 23.2, 26.1 and 26.8 ± 0.2° 2θ.

8. The crystalline form 1 of compound A-HCl according to claim 4, having an XRPD pattern substantially as shown in Figure 1.

9. The crystalline form 1 of compound A-HCl according to claim 4, having an endothermic transition from 268.5 °C to 274.5 °C when measured by differential scanning calorimetry.

10. The crystalline form 1 of compound A-HCl according to claim 9, wherein the endothermic transition is at 271.5 °C ± 3 °C.

11. The crystalline form 1 of compound A-HCl according to claim 10, having a thermogravimetric analysis (TGA) substantially as shown in Figure 2.

12. The crystalline form 1 of compound A-HCl according to claim 4, having a single crystal structure substantially as shown in Figure 5.

13. The hydrochloride salt of compound A according to claim 2, having the following structure: 【Chemical 2】

14. A pharmaceutical composition comprising the solid form of compound A-HCl according to claim 2, or the HCl salt of compound A according to claim 13, and a pharmaceutically acceptable excipient.

15. A method of treating a subject suffering from a disease mediated by KIF18A inhibition, the method comprising administering to a subject in need thereof a pharmaceutically effective amount of the pharmaceutical composition according to claim 14.

16. The method according to claim 15, wherein the disease mediated by the inhibition of KIF18A is a cancer selected from ovarian cancer, breast cancer, lung cancer, or endometrial cancer.

17. The method according to claim 15, wherein the subject is recurrent or refractory to at least one line of systemic chemotherapy.

18. The method according to claim 16, wherein the cancer is positive for an inactivated TP53 gene and / or an inactivated Rb gene, (ii) an amplified CCNE1 gene or an overexpressed CCNE1 gene product, (iii) an inactivated BRCA gene, or (iv) at least one of their combinations.

19. A method for preparing the compound A-HCl salt according to claim 2 or a solid form thereof, the method comprising combining hydrochloric acid, compound A and a suitable solvent to form the compound A-HCl or a solid form thereof.

20. The method according to claim 19, wherein the suitable solvent is selected from acetonitrile / water, acetonitrile / 1,4-dioxane, tetrahydrofuran / water, N-methyl-2-pyrrolidone / ethanol or acetone / water.

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