Polymorphisms of hm30181 mesylate

Abstract

To provide: a composition comprising polymorphs of HM30181 capable of improving absorption, improving pharmacokinetics, and / or reducing side effects upon administration; a method for treating cancer; and a formulation.SOLUTION: One embodiment of the present invention is a composition comprising a crystalline or partially crystalline form of HM30181 mesylate, wherein the crystalline or partially crystalline form includes polymorph B, polymorph C, polymorph D, polymorph E, polymorph F, polymorph G, polymorph H, polymorph I, polymorph J, polymorph K, polymorph L, polymorph M, and / or polymorph N.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The field of the invention is P-glycoprotein inhibitors, in particular HM30181 mesylate. [Background technology]

[0002] The background discussion includes information that may be useful in understanding the present invention. No admission is made that any of the information provided herein is prior art or relevant to the presently claimed invention, nor is any admission made that any publication referenced specifically or implicitly is prior art.

[0003] P-glycoprotein (P-gp) is an ATP-dependent efflux pump protein with broad substrate specificity found in major genera. Due to its widespread distribution and function, P-gp is thought to be a defense mechanism that actively transports toxins out of cells. In humans, P-gp can transport substrate compounds from intestinal epithelial cells back into the intestinal lumen, across the blood-brain barrier to adjacent capillaries, from the proximal tubule of the kidney to the urinary filtrate, and from hepatocytes to the bile duct.

[0004] Unfortunately, several drugs utilized in chemotherapy are substrates of P-gp. P-gp activity can therefore reduce the bioavailability and efficacy of chemotherapy drugs. In such instances, administration of P-gp inhibitors can be useful to improve response to chemotherapy. Accordingly, over the past 30 years, several pharmaceutically useful P-gp inhibitors have been developed, including amiodarone, clarithromycin, cyclosporine, colchicine, diltiazem, erythromycin, felodipine, ketoconazole, lansoprazole, omeprazole, nifedipine, paroxetine, reserpine, saquinavir, sertraline, quinidine, tamoxifen, verapamil, and duloxetine.

[0005] HM30181 mesylate is a third-generation P-gp inhibitor that has been investigated for use with paclitaxel. HM30181 mesylate selectively inhibits P-gp in the intestinal epithelium, improving the absorption of orally administered chemotherapy drugs without increasing potentially harmful transport across the blood-brain barrier. The structure of HM30181 mesylate is shown below.

[0006] [ka]

[0007] The pharmacokinetics, bioavailability, and incidence of side effects of orally administered HM30181 mesylate, however, are suboptimal. Thus, there remains a need for polymorphs of HM30181 that may provide improved absorption, improved pharmacokinetics, and / or reduced side effects upon administration. Summary of the Invention

[0008] The present subject matter provides polymorphs of HM31081, methods for their preparation and characterization, and methods for their use. One embodiment of the inventive concept is a composition comprising a crystalline or partially crystalline form of HM30181 mesylate, wherein the crystalline or partially crystalline form comprises polymorph B, polymorph C, polymorph D, polymorph E, polymorph F, polymorph G, polymorph H, polymorph I, polymorph J, polymorph K, polymorph L, polymorph M, and / or polymorph N. In some such embodiments, the crystalline or partially crystalline form is polymorph B, having an X-ray diffraction pattern corresponding to Figure 40 and an endotherm at about 159.92°C. In some such embodiments, the crystalline or partially crystalline form is polymorph C, having an X-ray diffraction pattern corresponding to Figure 42 and an endotherm at about 159.6°C. In some such embodiments, the crystalline or partially crystalline form is polymorph C, having an X-ray diffraction pattern with 2-theta maxima at about 6.4° and about 8.0°. In some of such embodiments, the crystalline or partially crystalline form is the Form C polymorph, having a crystallinity of about 1.180 nm. 3 (1180Å 3), where a is about 0.7 nm (7 Å), b is about 1.5 nm (15 Å), c is about 1.8 nm (18 Å), α is about 52°, β is about 62°, and γ is about 90°. In some such embodiments, the crystalline or partially crystalline form is polymorph C, having an X-ray diffraction pattern corresponding to Figure 42 and an endotherm at about 159.60°C, and may be a monohydrate. In some such embodiments, the crystalline or partially crystalline form is polymorph D, having an X-ray diffraction pattern corresponding to Figure 45 and an endotherm at about 66.97°C. In some such embodiments, the crystalline or partially crystalline form is polymorph E, having an X-ray diffraction pattern corresponding to Figure 47 and an endotherm at about 154.42°C, and may include DMA. In some such embodiments, the crystalline or partially crystalline form is the Form E polymorph and has an X-ray diffraction pattern corresponding to Figure 47 and an endotherm at about 154.4°C. In some such embodiments, the crystalline or partially crystalline form is the Form E polymorph and has an X-ray diffraction pattern with 2θ maxima at about 4.2°, about 10.4°, about 10.7°, about 14.7°, about 16.8°, about 21°, about 23.8°, about 26.6°, and about 27.7°. In some such embodiments, the crystalline or partially crystalline form is the Form E polymorph and has an X-ray diffraction pattern with 2θ maxima at about 1.620 nm 3 (1620Å 3), where a is about 0.8 nm (8 Å), b is about 1.0 nm (10 Å), c is about 2.4 nm (24 Å), α is about 75°, β is about 80°, and γ is about 110°. In some such embodiments, the crystalline or partially crystalline form is polymorph F, has an X-ray diffraction pattern corresponding to Figure 50 and an endotherm at about 148.41°C, and can include DMF. In some such embodiments, the crystalline or partially crystalline form is polymorph G, has an X-ray diffraction pattern corresponding to Figure 53 and an endotherm at about 69.02°C. In some such embodiments, the crystalline or partially crystalline form is polymorph H, has an X-ray diffraction pattern corresponding to Figure 55 and an endotherm at about 126.52°C. In some such embodiments, the crystalline or partially crystalline form is polymorph I and has an X-ray diffraction pattern corresponding to Figure 57. In some such embodiments, the crystalline or partially crystalline form is polymorph J and has an X-ray diffraction pattern corresponding to Figure 58. In some such embodiments, the crystalline or partially crystalline form is polymorph K and has an X-ray diffraction pattern corresponding to Figure 60. In some such embodiments, the crystalline or partially crystalline form is polymorph L and has an X-ray diffraction pattern corresponding to Figure 61. In some such embodiments, the crystalline or partially crystalline form is polymorph M and has an X-ray diffraction pattern corresponding to Figure 62. In some such embodiments, the crystalline or partially crystalline form is polymorph N and has an X-ray diffraction pattern corresponding to Figure 63 and endotherms at about 159°C and about 188°C, and may include methanol.

[0009] Another embodiment of the inventive concept is a method of inhibiting P-glycoprotein activity by contacting P-glycoprotein with one or more crystalline or partially crystalline forms of HM30181 mesylate as described above in an amount effective to inhibit the activity of P-glycoprotein.

[0010] Another embodiment of the inventive concept is a method of treating cancer by administering to an individual in need of cancer treatment a chemotherapeutic agent that is a substrate of P-glycoprotein, and further administering a polymorph of HM30181 mesylate, as described above, in an amount effective to inhibit P-glycoprotein activity in said individual.

[0011] Another embodiment of the inventive concept is the use of a polymorph of HM30181 mesylate as described above in the preparation of a medicament for the treatment of cancer, which may further comprise a chemotherapeutic agent that is a substrate of P-glycoprotein.

[0012] Another embodiment of the inventive concept is a formulation comprising a polymorph of HM30181 mesylate as described above and a therapeutic agent, wherein the therapeutic agent is a substrate of P-glycoprotein. Such a therapeutic agent can be a chemotherapeutic agent used in the treatment of cancer.

[0013] Various objects, features, aspects and advantages of the present subject matter may become more apparent from the following detailed description of preferred embodiments, taken in conjunction with the accompanying drawings in which like numerals represent like elements. [Brief explanation of the drawings]

[0014] [Figure 1] FIG. 1 shows an overlay of powder X-ray diffraction (XPRD) results for polymorphs A to N of HM30181 mesylate. [Figure 2] FIG. 1 shows the results of an XRPD test of Form A of HM30181 mesylate (starting material). [Figure 3] FIG. 1 shows an overlay of DSC / TGA test results for Form A of HM30181 mesylate. [Figure 4] FIG. 1 shows the results of a 1H-NMR test of Form A of HM30181 mesylate. [Figure 5] FIG. 1 shows the results of DVS for HM30181 mesylate type A (starting material). [Figure 6] Figure 1 shows an overlay of an XRPD run of Form A (starting material) after DVS. [Figure 7] FIG. 1 shows an overlay of cyclic DSC results for Form A HM30181 mesylate and Form A HM30181 mesylate heated to 110° C. [Figure 8] FIG. 1 shows an overlay of XRPD results for Form A of HM30181 mesylate and Form A of HM30181 mesylate heated to 110° C. [Figure 9] Figure 1 shows an overlay of cyclic DSC tests of HM30181 Form A mesylate (starting material) and HM30181 Form A mesylate heated to 185°C. [Figure 10] Figure 1 shows an overlay of XRPD experiments of Form A HM30181 mesylate (starting material) and Form A HM30181 mesylate heated to 185°C. [Figure 11] FIG. 1 shows an overlay of cyclic DSC results for Form A HM30181 mesylate (starting material) and Form A HM30181 mesylate heated to 200° C. [Figure 12] FIG. 1 shows an overlay of XRPD results for Form A HM30181 mesylate (starting material) and Form A HM30181 mesylate heated to 200° C. [Figure 13] FIG. 1 shows the results of an XRPD test of free base HM3018-A. [Figure 14] 1 shows an overlay of 1H-NMR results for Form A HM30181 mesylate heated to 200°C, free base HM3018-A, and Form A HM30181 mesylate (starting material). [Figure 15] Figure 1 shows an overlay of XRPD results for Form B HM30181 mesylate salt from a slurry experiment. [Figure 16] Figure 1 shows an overlay of XRPD results for Form C of HM30181 mesylate salt from a slurry experiment. [Figure 17] Figure 1 shows an overlay of XRPD results for Form D HM30181 mesylate salt from a slurry experiment. [Figure 18] Figure 1 shows an overlay of XRPD results for Form E of HM30181 mesylate salt from a slurry experiment. [Figure 19] Figure 1 shows an overlay of XRPD results for Form F HM30181 mesylate from a slurry experiment. [Figure 20] Figure 1 shows an overlay of XRPD results for Form G of HM30181 mesylate from a slurry experiment. [Figure 21] Figure 1 shows an overlay of XRPD results for Form D HM30181 mesylate from liquid vapor diffusion experiments. [Figure 22] Figure 1 shows an overlay of XRPD results for Form E HM30181 mesylate from liquid vapor diffusion experiments. [Figure 23] Figure 1 shows an overlay of XRPD results for Form H HM30181 mesylate from liquid vapor diffusion experiments. [Figure 24] FIG. 1 shows an overlay of XRPD results for Form I HM30181 mesylate and Form J HM30181 mesylate from liquid vapor diffusion experiments. [Figure 25] Figure 1 shows an overlay of XRPD results for the HM30181 mesylate salt of the Form C polymorph formed by cooling. [Figure 26] Figure 1 shows an overlay of XRPD results for Form C HM30181 mesylate and a sample from an antisolvent experiment. [Figure 27] Figure 1 shows an overlay of XRPD results for Form F HM30181 mesylate and a sample from an antisolvent experiment. [Figure 28] Figure 1 shows an overlay of XRPD results for J-form HM30181 mesylate and a sample from an antisolvent experiment. [Figure 29] Figure 1 shows an overlay of XRPD results for K-form HM30181 mesylate and a sample from an antisolvent experiment. [Figure 30] FIG. 1 shows an overlay of XRPD results for HM30181 Form L mesylate and a sample from an antisolvent experiment. [Figure 31]Figure 1 shows an overlay of XRPD results for M-form HM30181 mesylate and a sample from an antisolvent experiment. [Figure 32] Figure 1 shows an overlay of XRPD results for Form B HM30181 mesylate salt from a larger scale study. [Figure 33] Figure 1 shows an overlay of XRPD results for Form C of HM30181 mesylate from a larger scale study. [Figure 34] Figure 1 shows an overlay of XRPD studies of Form D HM30181 mesylate from a larger scale study. [Figure 35] Figure 1 shows an overlay of XRPD results for Form E HM30181 mesylate salt from a larger scale study. [Figure 36] Figure 1 shows an overlay of XRPD results for Form G HM30181 mesylate salt from a larger scale study. [Figure 37] Figure 1 shows an overlay of XRPD results for Form F HM30181 mesylate and Form G HM30181 mesylate from a larger scale study. [Figure 38] Figure 1 shows an overlay of 1H-NMR results of Form G HM30181 mesylate from a larger scale study. [Figure 39] Figure 1 shows an overlay of 1H-NMR results of the HM30181 mesylate polymorphs from a large scale study. [Figure 40] FIG. 1 shows the results of an XRPD test of Form B of HM30181 mesylate. [Figure 41] FIG. 1 shows an overlay of DSC and TGA results for Form B of HM30181 mesylate. [Figure 42] FIG. 1 shows the results of XRPD of Form C of HM30181 mesylate. [Figure 43] FIG. 1 shows an overlay of DSC and TGA results for Form C of HM30181 mesylate. [Figure 44] FIG. 1 shows the results of 1H-NMR of form C of HM30181 mesylate. [Figure 45] FIG. 1 shows the results of XRPD of form D of HM30181 mesylate. [Figure 46] FIG. 1 shows an overlay of DSC and TGA results for Form D of HM30181 mesylate. [Figure 47] FIG. 1 shows the results of XRPD of E-form HM30181 mesylate. [Figure 48] FIG. 1 shows an overlay of DSC and TGA results for Form E of HM30181 mesylate. [Figure 49] FIG. 1 shows the results of 1H-NMR of E-form HM30181 mesylate. [Figure 50] FIG. 1 shows the results of XRPD of form F of HM30181 mesylate. [Figure 51] FIG. 1 shows an overlay of DSC and TGA results for Form F of HM30181 mesylate. [Figure 52] FIG. 1 shows the results of 1H-NMR of F-form HM30181 mesylate. [Figure 53] FIG. 1 shows the results of XRPD of G-form HM30181 mesylate. [Figure 54] FIG. 1 shows an overlay of DSC and TGA results for form G of HM30181 mesylate. [Figure 55] FIG. 1 shows the results of XRPD of H-form HM30181 mesylate. [Figure 56] FIG. 1 shows an overlay of DSC and TGA results for Form H of HM30181 mesylate. [Figure 57] FIG. 1 shows the results of XRPD of Form I HM30181 mesylate. [Figure 58] FIG. 1 shows the results of XRPD of J-form HM30181 mesylate. [Figure 59] FIG. 1 shows the results of TGA of J-form HM30181 mesylate. [Figure 60] FIG. 1 shows the results of XRPD of K-form HM30181 mesylate. [Figure 61] FIG. 1 shows the results of XRPD of HM30181 form L mesylate. [Figure 62] FIG. 1 shows the results of XRPD of M-type HM30181 mesylate. [Figure 63] FIG. 1 shows the results of XRPD of N-type HM30181 mesylate. [Figure 64] FIG. 1 shows an overlay of DSC and TGA results for N-type HM30181 mesylate. [Figure 65] FIG. 1 shows the results of 1H-NMR of N-type HM30181 mesylate. DETAILED DESCRIPTION OF THE INVENTION

[0015] The following description contains information that may be useful in understanding the present invention. No admission is made that any of the information provided herein is prior art or relevant to the presently claimed invention, nor is any admission made that any publication referenced specifically or implicitly is prior art.

[0016] The present invention provides a wide range of polymorphs of HM30181 mesylate and methods for their preparation. The various polymorphs are structurally distinct as determined by X-ray diffraction and various physical properties. Polymorphs of HM30181 mesylate with improved pharmacokinetics, reduced incidence of side effects, reduced dosing regimens, etc. can be identified among them by conventional methods (e.g., animal studies, clinical trials, etc.).

[0017] It will be appreciated that the disclosed technology provides many advantageous technical effects, including improving the absorption of chemotherapy drugs while maintaining the patency of the blood-brain barrier, and reducing the incidence of the development of drug resistance during cancer treatment.

[0018] The following description provides many example embodiments of the inventive subject matter. Although each embodiment represents a single combination of inventive elements, it is contemplated that the inventive subject matter includes all possible combinations of the disclosed elements. Thus, if one embodiment comprises elements A, B, and C, and a second embodiment comprises elements B and D, then it is contemplated that the inventive subject matter further includes any other remaining combination of A, B, C, or D, even if not explicitly disclosed.

[0019] In some embodiments, numbers expressing properties such as amounts and concentrations of ingredients, reaction conditions, and the like, used to describe and claim particular embodiments of the invention should be understood to be modified in some instances by the term "about." Accordingly, in some embodiments, the numerical parameters set forth in the written description and appended claims are approximations that may vary depending on the properties desired to be obtained by a particular embodiment. In some embodiments, the numerical parameters should be construed in light of the number of reported significant digits and by applying ordinary rounding techniques. Notwithstanding that the numerical ranges and parameters setting forth the broad scope of some embodiments of the invention are approximations, the numerical ranges setting forth specific examples are reported as precisely as practicable. The numerical values ​​presented in some embodiments of the invention may contain certain errors necessarily resulting from the standard deviation found in their respective testing measurements.

[0020] As used throughout this description and the claims that follow, the meanings of "a / an" and "the" include plural references unless the context clearly dictates otherwise. Further, as used throughout this description, the meaning of "in" includes "in" and "on," unless the context clearly dictates otherwise.

[0021] The recitation of ranges of values ​​herein is merely intended to serve as a shorthand method of referring individually to each separate value falling within the range. Unless otherwise indicated herein, each separate value is incorporated herein as if it were individually recited herein. Any and all methods described herein can be performed in any suitable order unless otherwise indicated herein or clearly contradicted by context. The use of any and all examples or exemplary language (e.g., "such as") provided with respect to specific embodiments herein is intended solely to facilitate a better understanding of the invention and does not pose a limitation on the scope of the invention except as so stated. No language herein should be construed as indicating any unmentioned element essential to the practice of the invention.

[0022] Groupings of alternative elements or embodiments of the invention disclosed herein are not to be construed as limitations. The members of each group may be referred to and claimed individually or in any combination with other members of the group or other elements found herein. One or more members of a group may be included in, or deleted from, a group for reasons of convenience and / or patentability. When any such inclusion or deletion occurs, the specification is deemed to include the group as modified, thereby fulfilling all Markush group descriptions used in the appended claims.

[0023] The inventors have identified several polymorphs of HM30181 mesylate, including Forms A through N (see Table 1). Some of the polymorphs may include metastable solvates. The inventors believe that one or more of these polymorphs may have improved pharmacokinetics and / or bioavailability compared to prior art formulations of HM30181 mesylate. The inventors believe that such improvements may allow for the use of lower doses, reducing or eliminating side effects associated with treatment using prior art formulations of HM30181 mesylate.

[0024] [Table 1]

[0025] FIG. 1 provides a schematic overlay of powder X-ray diffraction (XPRD) results for the Forms A through N polymorphs of HM30181 mesylate, as provided in Table 1. Prior art HM30181 mesylate monohydrate (i.e., starting material) can be synthesized as described in WO 2005 / 033094 or U.S. Pat. No. 9,283,218, the entire contents of which are incorporated herein by reference. Such starting material was characterized by XRPD, TGA, DSC, and DVS (see below) and identified as crystalline Form A by XRPD (see FIG. 2).

[0026] By DSC, Form A HM30181 mesylate exhibited an endotherm at 181.41°C (see Figure 3). By TGA, Form A HM30181 mesylate exhibited a weight loss of 2.584% (HM30181 mesylate monohydrate with a MW of 802.849, a 2.24% loss) corresponding to the weight loss of the monohydrate before 150°C, followed by a weight loss of 4.133% likely due to dissociation and decomposition before 250°C (see Figure 3). 1 H-NMR showed that Form A of HM30181 mesylate was potentially a monohydrate, since no solvent other than water was detected (see Figure 4).

[0027] According to DVS, Form A of HM30181 mesylate was hygroscopic, absorbing 2.26% water at 0%-95% RH, and no changes in the XRPD pattern were observed (see Figures 5 and 6).

[0028] Cycling DSC of Form A of HM30181 mesylate to 110°C produced no change in XRPD (see Figures 7 and 8). Cycling DSC of Form A of HM30181 mesylate to 185°C produced Form A of HM30181 mesylate with reduced crystallinity (see Figures 9 and 10). Cycling DSC of Form A of HM30181 mesylate to 200°C produced amorphous material (see Figures 11 and 12). The free base, HM3018-A (6004273-06-A), was further characterized by XRPD and found to be amorphous (see Figure 13). 1 Based on 1 H-NMR data, decomposition of Form A began after heating to 200° C. (See Figure 14).

[0029] The solubility of HM30181 mesylate (starting material) was evaluated in several solvents (see below), and the results are listed in Table 2.

[0030] [Table 2]

[0031] Slurry-based generation and / or screening of polymorphs of HM30181 mesylate was carried out by preparing slurries of Form A of HM30181 mesylate (starting material) in various solvents and under various conditions, as described below. The resulting solids were analyzed by XRPD to identify the physical state. The results are summarized in Tables 3 and 4.

[0032] Slurrying Form A of HM30181 mesylate in MeOH at temperatures between 4°C and 50°C after one week produced Form B of HM30181 mesylate (see Figure 15). Slurrying Form A of HM30181 mesylate in DCM at ambient temperature and in acetonitrile at 4°C and 50°C produced Form C of HM30181 mesylate (see Figure 16). Slurrying Form A of HM30181 mesylate in NMP at ambient temperature produced Form D of HM30181 mesylate (see Figure 17). Both Form C and Form D of HM30181 mesylate showed significant similarity to Form A of HM30181 mesylate. Slurrying Form A of HM30181 mesylate in DMA at ambient conditions produced Form E of HM30181 mesylate (see Figure 18). Slurrying Form A of HM30181 mesylate in DMF produced Form F of HM30181 mesylate at ambient temperature (see Figure 19) and Form G at 50°C (see Figure 20).

[0033] [Table 3]

[0034] JPEG2025165973000005.jpg113170

[0035] [Table 4]

[0036] JPEG2025165973000007.jpg87170

[0037] Further generation and / or screening of polymorphs of HM30181 mesylate was performed by preparing Form A of HM30181 mesylate (starting material) for liquid and solid vapor diffusion, as described below. The resulting solid was analyzed by XRPD to identify the physical state. The results are summarized in Tables 5, 6, and 7. Form D of HM30181 mesylate was obtained by liquid vapor diffusion of MTBE into NMP solution (see Figure 21). Form D of HM30181 mesylate was obtained by liquid vapor diffusion of MEK into NMP or DMA solution (see Figure 21). Air-dried Form D of HM30181 mesylate showed some loss of crystallinity, suggesting a possible solvate. Form E of HM30181 mesylate was obtained by liquid vapor diffusion of 2-MeTHF into DMA or DMF solution (see Figure 22). Liquid vapor diffusion of MTBE into DMA solution gave Form E of HM30181 mesylate, which has several additional diffraction peaks that the inventors attribute to Form I of HM30181 mesylate (Error! Reference not found). Liquid vapor diffusion of ACN into DMSO solution gave Form H of HM30181 mesylate (see Figure 23). Liquid vapor diffusion of acetone, ethyl acetate, or isopropyl acetate into DMA solution gave Form I of HM30181 mesylate (see Figure 24). Air-drying of Form I of HM30181 mesylate gave Form J of HM30181 mesylate (see Figure 24). Liquid vapor diffusion of isopropyl acetate into DMSO solution followed by air-drying further gave Form J of HM30181 mesylate.

[0038] [Table 5]

[0039] [Table 6]

[0040] JPEG2025165973000010.jpg217170

[0041] JPEG2025165973000011.jpg130170

[0042] [Table 7]

[0043] Generation and / or screening of polymorphs of HM30181 mesylate by cooling was carried out by treating Form A of HM30181 mesylate (starting material) with slow or rapid cooling (i.e., crash cooling), as described below. The resulting solid was analyzed by XRPD to identify the physical state. The results are summarized in Table 8. Cooling experiments in acetonitrile and DCM yielded Form C of HM30181 mesylate, and air drying did not show any significant change (see Figure 25).

[0044] [Table 8]

[0045] HM30181 mesylate was further subjected to the evaporation method by treating Form A of HM30181 mesylate (starting material) as described below. The resulting solid was analyzed by XRPD to identify its physical state. The results are shown in Table 9.

[0046] [Table 9]

[0047] The generation and / or screening of polymorphs of HM30181 mesylate by treatment with anti-solvents was carried out by treating Form A of HM30181 mesylate (starting material) as described below. The resulting solid was analyzed by XRPD to identify the physical state. The results are summarized in Tables 10, 11, 12, and 13. An anti-solvent study in DMSO gave Form C of HM30181 mesylate (see Figure 26). An anti-solvent study in N,N-dimethylacetamide with methyl t-butyl ether gave Form F of HM30181 mesylate (see Figure 27). Anti-solvent addition and reverse anti-solvent addition in DMSO / EtOAc, DMA / MIBK, DMA / toluene, and NMP / t-BuOH gave a primarily amorphous fraction and one polymorph that shares some similarity with Form J of HM30181 mesylate (see Figure 28). Further anti-solvent experiments in DMSO and DMF yielded the K polymorph of HM30181 mesylate (or possibly a mixture of forms, see Figure 29). Anti-solvent experiments in DMF / n-propanol and DMA / isopropanol yielded the L polymorph of HM30181 mesylate (see Figure 30). Anti-solvent additions of DMF / toluene and DMA / t-BuOH were mostly amorphous but also produced Form M of HM30181 mesylate (see Figure 31).

[0048] [Table 10]

[0049] JPEG2025165973000016.jpg26170

[0050] [Table 11]

[0051] JPEG2025165973000018.jpg24170

[0052] [Table 12]

[0053] JPEG2025165973000020.jpg24170

[0054] [Table 13]

[0055] Larger scale studies were performed on a 200 mg scale using Form A HM30181 mesylate (starting material) as described below. The results are summarized in Table 14. The solid was isolated by vacuum filtration. The filtered wet cake from the DMA, DMF, and NMP slurry was washed with 1 mL to 2 mL of methanol to remove the solvent. The solid was then vacuum dried at 80°C overnight.

[0056] On a larger scale, a slurry of Form A of HM30181 mesylate (starting material) in methanol at ambient conditions yielded a mixture of Form B of HM30181 mesylate and Form A of HM30181 mesylate after 9 days (see Figure 32). After 14 days, a slurry in methanol yielded the N-type polymorph of HM30181 mesylate. Form N of HM30181 mesylate showed some loss of crystallinity after vacuum drying, suggesting it is a methanol solvate (see Figure 32). A slurry of Form A of HM30181 mesylate in acetonitrile (starting material) at ambient conditions yielded Form C of HM30181 mesylate after 14 days, and no loss of crystallinity was detected after vacuum drying (see Figure 33). A scaled-up slurry of Form A of HM30181 mesylate (starting material) in NMP provided primarily amorphous material (see Figure 34). A scaled-up slurry of Form A of HM30181 mesylate (starting material) in DMA gave Form E of HM30181 mesylate after 6 days (see Figure 35). Form E of HM30181 mesylate showed no loss of crystallinity after vacuum drying. A scaled-up slurry of Form A of HM30181 mesylate (starting material) in DMF at 50°C gave Form F of HM30181 mesylate after 9 days (see Figure 36), rather than the expected Form G of HM30181 mesylate. Some change in the pattern was observed after vacuum drying. A scaled-up slurry of Form A of HM30181 mesylate (starting material) in DMF at ambient temperature initially showed no change from Form A (see Figure 37). Such a slurry was heated in an attempt to produce Form G of HM30181 mesylate. Scaled-up slurrying of Form A HM30181 mesylate (starting material) in DMF at 100°C gave Form F HM30181 mesylate after 2 days (see Figure 37). Scaled-up slurrying of Form A HM30181 mesylate (starting material) in DMF at 150°C gave a previously unobserved XRPD pattern after 5 days (see Figure 37). 1 The H-NMR results show 1 The 1 H-NMR spectrum did not correspond to either the starting material or the free base, suggesting decomposition of the DMF slurry at 150° C. (See Figure 38).

[0057] 1 No disproportionation or decomposition was detected in the C, E, F, and N polymorphs of HM30181 mesylate by 1 H-NMR (see Figure 39). 1 H-NMR revealed that form E of HM30181 mesylate contained DMA, form F of HM30181 mesylate contained DMF, and form N of HM30181 mesylate contained MeOH.

[0058] [Table 14]

[0059] The polymorphic properties of HM30181 mesylate, characterized by XRPD, TGA, DSC and DVS, are summarized below: Form A of HM30181 mesylate represents a prior art preparation of HM30181 mesylate that can be used as a starting material in the generation of novel polymorphs of such compounds.

[0060] Crystalline Form B of HM30181 mesylate was obtained by slurrying in methanol at 4°C to 50°C and characterized by XRPD (see Figure 40). By DSC, Form B of HM30181 mesylate exhibited an endotherm at 159.92°C (see Figure 41). By TGA, Form B of HM30181 mesylate exhibited a weight loss of 1.865% before 170°C, followed by possible dissociation and decomposition (see Figure 41).

[0061] The crystalline Form C polymorph of HM30181 mesylate was obtained by slurrying in acetonitrile at ambient temperature. Form C of HM30181 mesylate exhibited distinctive XRPD results. After overnight vacuum drying, no change in the crystallinity pattern was observed (see Figure 42). DSC revealed that Form C of HM30181 mesylate exhibited an endotherm at 159.60°C (see Figure 43). TGA revealed that Form C of HM30181 mesylate exhibited a 2.987% weight loss before 170°C, followed by dissociation and decomposition (see Figure 43). 1H-NMR confirmed that Form C of HM30181 mesylate contained only water and no acetonitrile (~2.07 ppm), suggesting that it is a monohydrate (see Figure 44).

[0062] Crystalline Form D HM30181 mesylate was obtained by slurrying in N-methylpyrrolidone at ambient temperature, and exhibits characteristic XRPD results. After air-drying overnight, no change in the crystalline pattern was observed (see Figure 45). By DSC / TGA, Form D HM30181 mesylate exhibited an endotherm at 66.97°C and a weight loss of 14.71% before 150°C, suggesting significant residual solvent content (see Figure 46).

[0063] Crystalline Form E of HM30181 mesylate was obtained by slurrying in N,N-dimethylacetamide at ambient temperature, and exhibited characteristic XRPD results. After overnight vacuum drying at 80°C, no change in the crystalline pattern was observed (see Figure 47). By DSC / TGA, Form E of HM30181 mesylate exhibited an endotherm at 154.42°C and a 5.247% weight loss before 168°C, suggesting that Form E of HM30181 mesylate is a solvate (see Figure 48). 1 H-NMR confirmed that Form E of HM30181 mesylate contained DMA (see Figure 49).

[0064] Crystalline Form F of HM30181 mesylate was obtained by slurrying in dimethylformamide at 50°C, and exhibits characteristic XRPD results (see Figure 50). After drying overnight under vacuum at 80°C, a significant decrease in crystallinity was observed, suggesting that Form F of HM30181 mesylate is a metastable solvate. By DSC / TGA, Form F of HM30181 mesylate exhibited an endotherm at 148.41°C and a weight loss of 5.123% before 180°C, consistent with the loss of DMF (see Figure 51). 1 H-NMR confirmed that Form F of HM30181 mesylate contained DMF (see Figure 52).

[0065] Crystalline Form G of HM30181 mesylate was obtained by slurrying in dimethylformamide at 50°C. Form G of HM30181 mesylate exhibited distinctive XRPD results. After air-drying overnight, no decrease in crystallinity was observed (see Figure 53). DSC / TGA showed endotherms at 69.02°C and 233.29°C, and a weight loss of 1.451% before 150°C, consistent with the loss of DMF (see Figure 54).

[0066] Crystalline Form H of HM30181 mesylate was obtained by liquid vapor diffusion of acetonitrile into a DMSO stock of HM30181 mesylate, and Form H of HM30181 mesylate exhibits distinctive XRPD results (see Figure 55). By DSC / TGA, Form H of HM30181 mesylate exhibited an endotherm at 126.52°C and a weight loss of 7.444% before 200°C, which may be due to residual ACN and DMSO or a solvate (see Figure 56).

[0067] Crystalline Form I HM30181 mesylate was obtained by liquid vapor diffusion of acetone, ethyl acetate, or isopropyl acetate into a DMA stock of HM30181 mesylate, and Form I HM30181 mesylate exhibits characteristic XRPD results (see Figure 57). Air-drying of Form I HM30181 mesylate gave Form J HM30181 mesylate.

[0068] Form J of HM30181 mesylate was obtained by liquid vapor diffusion of acetone, ethyl acetate, or isopropyl acetate into a DMA stock of HM30181 mesylate, or by liquid vapor diffusion of isopropyl acetate into a DMSO stock of HM30181 mesylate, followed by air drying. By XRPD, Form J of HM30181 mesylate is crystalline (see Figure 58). By TGA, Form J of HM30181 mesylate exhibited a weight loss of 2.887% before 150°C, suggesting that it is a solvate or hydrate (see Figure 59).

[0069] Form K of HM30181 mesylate was obtained by anti-solvent addition using DMF / IPA and several DMSO systems (ethanol, acetone, MIBK, THF, chloroform, t-butanol, n-propyl acetate, and n-propanol). By XRPD, Form K of HM30181 mesylate is partially crystalline (see Figure 60).

[0070] Form L of HM30181 mesylate was obtained by anti-solvent addition in DMF / n-propanol and DMA / isopropanol systems, and was partially crystalline by XRPD (see Figure 61).

[0071] Form M of HM30181 mesylate was obtained by anti-solvent addition in DMF / toluene and DMA / t-butanol systems, and was partially crystalline by XRPD (see Figure 62).

[0072] Crystalline Form N of HM30181 mesylate was obtained after treating Form A of HM30181 mesylate (starting material) as a slurry in methanol at ambient temperature for 14 days (see Figure 63). Form N of HM30181 mesylate showed some loss of crystallinity after drying under vacuum, suggesting it is a methanol solvate. By DSC, Form N of HM30181 mesylate exhibited endotherms at 159.28 °C and 188.47 °C, with a weight loss of 2.128% before 180 °C, possibly indicating dissociation and decomposition (see Figure 64). 1 H-NMR confirmed the presence of methanol (see Figure 65).

[0073] Forms C and E of HM30181 mesylate were further analyzed to determine unit cell dimensions. Unit cell parameters for the Form C polymorph of HM30181 mesylate were calculated using the cumulative XRPD spectrum, with peak identifications shown in Table 15. Particularly distinct peaks for Form C of HM30181 mesylate are shown in bold and italics in Table 15. Estimates of the unit cell parameters derived from the Form C polymorph of HM30181 mesylate are shown in Table 16, and are consistent with a triclinic simple (P) unit cell.

[0074] [Table 15]

[0075] [Table 16]

[0076] Characteristic XRPD peak values ​​for the Form E polymorph are provided in Table 17, with particularly distinct peaks shown in bold and italics, which are understood to be distinct and different from those of the Form C polymorph, indicating that the Forms C and E polymorphs are distinct and different from each other, and that both the Forms C and E polymorphs are distinct and different from the prior art Form A polymorph of HM30181 mesylate salt.

[0077] [Table 17]

[0078] JPEG2025165973000026.jpg117170

[0079] The unit cell parameters for the E polymorphic HM30181 mesylate salt were estimated using cumulative XRPD spectra. The estimated unit cell parameters derived from the E polymorphic HM30181 mesylate salt are shown in Table 18 and are consistent with a triclinic simple (P) unit cell.

[0080] [Table 18]

[0081] As described above, HM30181 is an inhibitor of P-glycoprotein, an efflux transporter protein that is effective in removing a wide range of therapeutic substances from cells and forms an important part of the blood-brain barrier. While this function is essentially protective, it can adversely affect the use of therapeutic agents that are substrates of P-glycoprotein. Examples of drugs transported by P-glycoprotein include, but are not limited to, antineoplastic agents (e.g., docetaxel, etoposide, vincristine), calcium channel blockers (e.g., amlodipine), calcineurin inhibitors (e.g., cyclosporine, tacrolimus), digoxin, macrolide antibiotics (e.g., clarithromycin), and protease inhibitors. Therefore, HM30181 mesylate can be used to alter the pharmacokinetics of therapeutic agents that are substrates of P-glycoprotein by reducing the efflux of such drugs from multiple cells in an individual receiving treatment.

[0082] Conventional processes for the production of HM30181 provide the Form A polymorph. The inventors have produced and identified several other forms of such compounds, including Forms B, C, D, E, F, G, H, I, J, K, L, M, and N polymorphs of HM30181. As indicated above, these are different and distinct from the prior art Form A polymorph and from each other. The inventors believe that these new polymorphs of HM30181 may provide stability and / or pharmacokinetics (e.g., absorption rate, etc.) that differ from those provided by the prior art Form A polymorph.

[0083] Thus, another embodiment of the inventive concept is the application of one or more of the B, C, D, E, F, G, H, I, J, K, L, M, and / or N polymorphs of HM30181 to inhibit P-glycoprotein, thereby altering the pharmacokinetics of drugs that are substrates of P-glycoprotein. In some such embodiments, the drugs can be chemotherapeutic drugs used in cancer treatment.

[0084] In such embodiments, one or more of the B, C, D, E, F, G, H, I, J, K, L, M, and / or N polymorphs of HM30181 can be administered in conjunction with a drug that is a P-glycoprotein substrate to an individual in need of treatment for a disease or condition responsive to such a drug. In some embodiments, the B, C, D, E, F, G, H, I, J, K, L, M, and / or N polymorphs of HM30181 can be provided as separate formulations. Alternatively, one or more of the B, C, D, E, F, G, H, I, J, K, L, M, and / or N polymorphs of HM30181 can be formulated in combination with a drug that is a P-glycoprotein substrate. In a preferred embodiment, the disease is cancer and the agent that is a P-glycoprotein substrate is a chemotherapy agent used to treat cancer.

[0085] method As described above, multiple polymorphs of HM30181 mesylate were prepared by processing a conventional preparation of HM30181 mesylate Form A with various solvents and using various techniques. To test the solubility of HM30181 mesylate Form A in various solvents, a solid sample (∼2 mg) was transferred into a 4 mL glass vial. Solvent was added to the vial in 50 μL increments up to a total volume of 100 μL, followed by 100 μL increments until the concentration was less than 1.0 mg / mL. After each addition, the sample was thoroughly mixed by ultrasound for 2 minutes and vortexed for 1 minute. The solvent volumes (V1 and V2) were recorded and used to estimate solubility. The solvents used are summarized in Table 19 below.

[0086] [Table 19]

[0087] Screening for multiple polymorphs of HM30181 mesylate can involve preparing a slurry. Typically, a slurry was prepared by suspending 5 mg to 20 mg of sample in 0.1 mL to 0.5 mL of solvent in a 1.5 mL or 3.0 mL glass vial. The suspension was stirred at 200 rpm at the target temperature (e.g., 4°C, ambient temperature, 50°C). Solids for powder X-ray diffraction (XRPD) analysis were isolated by centrifugation at 14,000 rpm for 5 minutes at ambient temperature. If a solid or gel is not obtained, the slurry can be transferred to a fume hood for solvent evaporation.

[0088] In some embodiments, anti-solvent addition was used. In such a method, a concentrated stock of the compound in a solvent was prepared. The anti-solvent was then added quickly to the concentrated solution while stirring to induce precipitation. Filtration or centrifugation can be used to isolate the solid for XRPD analysis.

[0089] In some embodiments, reverse anti-solvent addition was used. In this method, a concentrated stock of the compound in a solvent is prepared and rapidly added to an anti-solvent with stirring to induce precipitation. Filtration or centrifugation can be used to isolate the solid for XRPD analysis.

[0090] In some embodiments, slow cooling was used. In this method, a concentrated suspension of the compound in a solvent was prepared. The solution was heated to 50°C and held at 50°C for at least 30 minutes. The resulting solution or suspension was filtered at 50°C using a 0.45 micron PTFE filter, and the filtrate was collected in a clean vial. The resulting clear solution was cooled to 5°C to induce precipitation. Filtration or centrifugation was used to separate the solid for XRPD analysis.

[0091] In some embodiments, rapid cooling was used. In such a method, a concentrated suspension of the compound in a solvent was prepared. The suspension was heated to 50°C and held at 50°C for at least 30 minutes. The heated solution or suspension was filtered at 50°C using a 0.45 micron PTFE filter, and the filtrate was collected in a clean vial. The clear solution was cooled to -20°C to induce precipitation. Filtration or centrifugation was used to separate the solid for XRPD analysis.

[0092] In some embodiments, liquid vapor diffusion was used. In this method, a concentrated suspension of the compound in a solvent is prepared. This concentrated stock is transferred to an inner vial that is sealed within a larger vial containing an antisolvent. Filtration or centrifugation is used to separate the solid for XRPD analysis.

[0093] In some embodiments, solid vapor diffusion was used. In such a method, 5-15 mg of sample was weighed into a small (e.g., 3 mL) vial. The vial was placed inside a larger (e.g., 20 mL) vial containing 3-4 mL of a volatile solvent. The outer vial was then sealed. The assembly was kept at ambient temperature for 7 days to allow the solvent vapor to interact with the solid, and the resulting product was characterized by XRPD.

[0094] The unique polymorphic HM30181 mesylate salt was characterized by a variety of techniques, including X-ray powder diffraction (XRPD), NMR, and calorimetry, as follows. XRPD was performed using a Panalytical X'Pert3® powder XRPD on a Si zero-background holder. 2θ positions were calibrated against a Panalytical® 640 Si powder standard. Details of the XRPD used in the experiments are listed in Table 20 below.

[0095] [Table 20]

[0096] Differential scanning calorimetry (DSC) was performed using a TA Instruments TA Q2000® DSC. With the pan crimped, the temperature was increased from ambient to the desired temperature at a heating rate of 10°C / min using N2 as the purge gas (see Table 21).

[0097] [Table 21]

[0098] Some tests used a cyclic DSC method in which the temperature was increased from ambient to 150° C. at a heating rate of 10° C. / min using N2 as a purge gas, followed by cooling from 10° C. to 25° C. Such a temperature cycle was repeated twice (see Table 22).

[0099] [Table 22]

[0100] Thermogravimetric analysis (TGA) was performed using a TA Instruments TA Q500® TGA. With the pan open, the temperature was increased from ambient to the desired temperature at a heating rate of 10°C / min using N2 as the purge gas (see Table 23).

[0101] [Table 23]

[0102] Dynamic Vapor Sorption (DVS) was measured using an SMS (Surface Measurement Systems®) DVS Intrinsic. The parameters for the DVS test are listed in Table 24 below.

[0103] [Table 24]

[0104] Proton NMR was obtained in deuterated DMSO (DMSO-d6) using a Varian 200M® NMR. It will be apparent to those skilled in the art that numerous modifications beyond those already described are possible without departing from the inventive concepts herein. The inventive subject matter is therefore not limited except in the spirit of the appended claims. Furthermore, in interpreting both the specification and the claims, all terms shall be understood in the broadest possible manner consistent with the context. In particular, the terms "comprises" and "comprising" shall be interpreted as referring to elements, components, or steps in a non-exclusive manner, indicating that a referenced element, component, or step may be present in, utilized with, or combined with other elements, components, or steps not specifically referenced. When a claim herein recites one or more of anything selected from the group consisting of A, B, C, and N, the document shall be interpreted as requiring only one element from the group, not A+N or B+N, etc.

Claims

1. 1. A composition comprising HM30181 mesylate in a crystalline or partially crystalline form, wherein the crystalline or partially crystalline form comprises one or more of polymorph B, polymorph C, polymorph D, polymorph E, polymorph F, polymorph G, polymorph H, polymorph I, polymorph J, polymorph K, polymorph L, polymorph M, and polymorph N.

2. 10. The composition of claim 1, wherein the crystalline or partially crystalline form is polymorph B, and the crystalline form has an X-ray diffraction pattern corresponding to Figure 40 and an endotherm at about 159.92°C.

3. 10. The composition of claim 1, wherein the crystalline or partially crystalline form is the Form C polymorph, and the crystalline form has an X-ray diffraction pattern corresponding to Figure 42 and an endotherm at about 159.6°C.

4. 10. The composition of claim 1, wherein the crystalline or partially crystalline form is the Form C polymorph, and the crystalline form has an X-ray diffraction pattern with 2-theta maxima at about 6.4° and about 8.0°.

5. The crystalline or partially crystalline form is the Form C polymorph, and the crystalline form has a molecular weight of about 1.180 nm. ３ (1180 Å ３ 2. The composition of claim 1, comprising a unit cell having a volume of about 0.7 nm (7 Å), b is about 1.5 nm (15 Å), c is about 1.8 nm (18 Å), α is about 52°, β is about 62, and γ is about 90°.

6. 7. The composition of claim 6, wherein the crystalline form comprises a monohydrate of HM30181 mesylate.

7. 10. The composition of claim 1, wherein the crystalline or partially crystalline form is polymorph D, and the crystalline form has an X-ray diffraction pattern corresponding to Figure 45 and an endotherm at about 66.97°C.

8. 10. The composition of claim 1, wherein the crystalline or partially crystalline form is polymorph E, and the crystalline form has an X-ray diffraction pattern corresponding to Figure 47 and an endotherm at about 154.42°C.

9. 10. The composition of claim 9, wherein the crystalline or partially crystalline form comprises DMA.

10. 10. The composition of claim 1, wherein the crystalline or partially crystalline form is the Form E polymorph, and the crystalline form has an X-ray diffraction pattern with 2θ maxima at about 4.2°, about 10.4°, about 10.7°, about 14.7°, about 16.8°, about 21°, about 23.8°, about 26.6°, and about 27.7°.

11. The crystalline or partially crystalline form is the E polymorph, and the crystalline form has a molecular weight of about 1.620 nm. ３ (1620 Å ３ ), a is about 0.8 nm (8 Å), b is about 1.0 nm (10 Å), c is about 2.4 nm (24 Å), α is about 75°, β is about 80°, and γ is about 110°.

12. 10. The composition of claim 1, wherein the crystalline or partially crystalline form is polymorph F, and the crystalline form has an X-ray diffraction pattern corresponding to Figure 50 and an endotherm at about 148.41°C.

13. 13. The composition of claim 12, wherein the crystalline or partially crystalline form comprises DMF.

14. 10. The composition of claim 1, wherein the crystalline or partially crystalline form is polymorph G, and the crystalline form has an X-ray diffraction pattern corresponding to Figure 53 and an endotherm at about 69.02°C.

15. 10. The composition of claim 1, wherein the crystalline or partially crystalline form is polymorph H, and the crystalline form has an X-ray diffraction pattern corresponding to Figure 55 and an endotherm at about 126.52°C.

16. 10. The composition of claim 1, wherein the crystalline or partially crystalline form is polymorph I, and the crystalline form has an X-ray diffraction pattern corresponding to Figure 57.

17. 10. The composition of claim 1, wherein the crystalline or partially crystalline form is polymorph J, and the crystalline form has an X-ray diffraction pattern corresponding to Figure 58.

18. 10. The composition of claim 1, wherein the crystalline or partially crystalline form is polymorph K, and the crystalline form has an X-ray diffraction pattern corresponding to Figure 60.

19. 10. The composition of claim 1, wherein the crystalline or partially crystalline form is polymorph L, and the crystalline form has an X-ray diffraction pattern corresponding to Figure 61.

20. 10. The composition of claim 1, wherein the crystalline or partially crystalline form is polymorph M, and the crystalline form has an X-ray diffraction pattern corresponding to Figure 62.

21. 10. The composition of claim 1, wherein the crystalline or partially crystalline form is polymorph N, and the crystalline form has an X-ray diffraction pattern corresponding to Figure 63 with endotherms at about 159°C and about 188°C.

22. 22. The composition of claim 21, wherein the crystalline or partially crystalline form comprises methanol.

23. 23. A method of inhibiting P-glycoprotein activity, comprising contacting P-glycoprotein with one or more crystalline or partially crystalline forms of HM30181 mesylate according to claims 1-22 in an amount effective to inhibit the activity of P-glycoprotein.

24. 1. A method of treating cancer, comprising: administering a chemotherapeutic agent to an individual in need of treatment; administering to the individual in need thereof a polymorph of HM30181 mesylate according to at least one of claims 1 to 22 in an amount effective to inhibit P-glycoprotein activity in said individual; The method wherein said chemotherapeutic agent is a substrate of P-glycoprotein.

25. Use of a polymorph of HM30181 mesylate according to at least one of claims 1 to 22 in preparing a medicament for the treatment of cancer.

26. 26. The use of claim 25, wherein the agent further comprises a chemotherapeutic agent that is a substrate of P-glycoprotein.

27. A formulation comprising the polymorph of HM30181 mesylate salt according to at least one of claims 1 to 22 and a therapeutic agent, wherein the therapeutic agent is a substrate of P-glycoprotein.

28. 28. The formulation of claim 27, wherein the therapeutic agent is a chemotherapy agent used in the treatment of cancer.

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