Patent ID: 12227494

DETAILED DESCRIPTION

The following description includes information that may be useful in understanding the present invention. It is not an admission that any of the information provided herein is prior art or relevant to the presently claimed invention, or that any publication specifically or implicitly referenced is prior art.

The inventive subject matter provides a wide range of polymorphisms of HM30181 mesylate and methods for their preparation. The various polymorphisms are shown to be structurally distinct by X-ray diffraction and various physical properties. Polymorphs of HM30181 mesylate with improved pharmacokinetics, reduced incidence of side effects, reduced dosing schedules, etc. can be identified among these by conventional methods (e.g., animal studies, clinical studies, etc.).

One should appreciate that the disclosed techniques provide many advantageous technical effects including improving absorption of chemotherapeutic drugs while maintaining patency of the blood-brain barrier and reducing the incidence of developing drug resistance during cancer treatment.

The following discussion provides many example embodiments of the inventive subject matter. Although each embodiment represents a single combination of inventive elements, the inventive subject matter is considered to include 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 the inventive subject matter is also considered to include other remaining combinations of A, B, C, or D, even if not explicitly disclosed.

In some embodiments, the numbers expressing quantities of ingredients, properties such as concentration, reaction conditions, and so forth, used to describe and claim certain embodiments of the invention are to be understood as being modified in some instances by the term “about.” Accordingly, in some embodiments, the numerical parameters set forth in the written description and attached claims are approximations that can vary depending upon the desired properties sought 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 values set forth in the 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.

As used in the description herein and throughout the claims that follow, the meaning of “a,” “an,” and “the” includes plural reference unless the context clearly dictates otherwise. Also, as used in the description herein, the meaning of “in” includes “in” and “on” unless the context clearly dictates otherwise.

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 individual value is incorporated into the specification as if it were individually recited herein. All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g. “such as”) provided with respect to certain embodiments herein is intended merely to better illuminate the invention and does not pose a limitation on the scope of the invention otherwise claimed. No language in the specification should be construed as indicating any non-claimed element essential to the practice of the invention.

Groupings of alternative elements or embodiments of the invention disclosed herein are not to be construed as limitations. Each group member can 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 can 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 herein deemed to contain the group as modified thus fulfilling the written description of all Markush groups used in the appended claims.

Inventors have identified 1a number of polymorphisms of HM30181 mesylate, including Type A to Type N (see Table 1). Some of the polymorphisms can include metastable solvates. The Inventors believe that one or more of these polymorphs can have improved pharmacokinetics and/or bioavailability relative to prior art formulations of HM30181 mesylate. Inventors believe that such improvements can permit the use of lower doses that reduce or eliminate side effects associated with treatment using prior art formulations of HM30181 mesylate.

TABLE 0CrystalDSCformendothermBatch(XRPD)(° C.)TGA weight lossIdentificationB00505-05-CType A181.412.6% before 150° C.Monohydrate (hygroscopic(starting material)to 2.26% at 95% RH)6007235-16-A1-airdryType B159.921.9% before 170° C.Solvate/hydrate6004273-10-C-vdType C159.603.0% before 175° C.Monohydrate6007235-16-A21-airdryType D66.97 (peak),14.71% before 150° C.Solvate/hydrate114 (exotherm)6004273-10-E-vdType E154.425.247% before 168° C.DMA solvate6004273-10-G-vdType F148.415.123% weight lossDMF solvatebefore 180° C.6007235-16-B26-airdryType G69.021.451% before 150° C.Solvate/hydrate233.296007235-19-B10Type H126.527.444% weight lossSolvate/hydratebefore 200° C.6007235-19-E3Type INANAWeak solvate- converts toType J on air-drying6007235-19-E5Type JNA2.887% weight lossSolvate/hydratebefore 150° C.6004273-06-A4Type KInsufficientInsufficient samplePossible solvate/hydratesample6004273-06-A60Type LInsufficientInsufficient samplePossible solvate/hydratesample6004273-06-A56Type MInsufficientInsufficient samplePossible solvate/hydratesample6004273-10-B-vdType N159.282.128% weight lossPossible MeOH188.47before 180° C.solvate/hydrate
FIG.1provides a summary overlay of X-Ray Powder Diffraction (XRPD) results from HM30181 mesylate polymorphisms types A to N, as provided in Table 1.

A prior art HM30181 mesylate salt monohydrate material (i.e., a starting material) can be synthesized as described in PCT application publication number WO 2005/033097 or U.S. Pat. No. 9,283,218 which are incorporated herein by reference. This starting material was characterized by XRPD, TGA, DSC, and DVS (see below), and was identified as crystalline Type A by XRPD (seeFIG.2).

By DSC, HM30181 mesylate Type A displayed an endotherm at 181.41° C. (seeFIG.3). By TGA, HM30181 mesylate Type A showed 2.584% weight loss before 150° C., matching with a monohydrate weight loss (MW 802.849 for HM30181 mesylate salt monohydrate, 2.24% wt loss), followed by a 4.133% weight loss before 250° C., possibly due to disassociation and decomposition (seeFIG.3). By1H-NMR, HM30181 mesylate Type A was potentially a hydrate, as no solvents were detected other than water (seeFIG.4).

By DVS, HM30181 mesylate Type A was hygroscopic and absorbed 2.26% water from 0-95% RH, with no change in XRPD pattern observed (seeFIG.5andFIG.6).

Cyclic DSC to 110° C. of HM30181 mesylate Type A resulted in no XRPD change (seeFIG.7andFIG.8). Cyclic DSC of HM30181 mesylate Type A to 185° C. resulted in HM30181 mesylate Type A with reduced crystallinity (FIG.9andFIG.10). Cyclic DSC of HM30181 mesylate Type A to 200° C. resulted in amorphous material (seeFIG.11andFIG.12). Freebase HM3018-A (6004273-06-A) was also characterized by XRPD and was found to be amorphous (seeFIG.13). Based on1H-NMR data, decomposition of Type A initiated after heating to 200° C. (seeFIG.14).

Solubility of HM30181 mesylate starting material was estimated in solvents (see below), and results are listed in Table 2.

TABLE 2Dissolved afterSolubilityheating to 50° C.Experiment IDSolvent(mg/mL)for 2 hours?B00505-05-C-1MeOH1.7-2.0B00505-05-C-2EtOH<0.9AlmostB00505-05-C-3IPA<1.0NoB00505-05-C-4Acetone<1.0NoB00505-05-C-5MIBK<1.0NoB00505-05-C-6EtOAc<0.9NoB00505-05-C-7IPOAc<1.0NoB00505-05-C-8THF<1.0NoB00505-05-C-92-MeTHF<1.0NoB00505-05-C-101,4-Dioxane<1.0NoB00505-05-C-11MTBE<1.0NoB00505-05-C-12ACN<1.0YesB00505-05-C-13DCM<1.0YesB00505-05-C-14CHCl35.0-6.7B00505-05-C-15Toluene<1.0NoB00505-05-C-16n-Heptane<1.0NoB00505-05-C-17H2O<1.0NoB00505-05-C-18Cyclohexane<1.0NoB00505-05-C-19MEK<1.0NoB00505-05-C-20T-BuOH<1.0NoB00505-05-C-21NMP4.8-6.3B00505-05-C-22DMSO22-44B00505-05-C-23DMA3.2-4.8B00505-05-C-24n-Propanol<1.0NoB00505-05-C-25n-Propyl acetate<1.0NoB00505-05-C-26Cumene<1.0No

Slurry-based generation and/or screening for HM30181 mesylate polymorphs was performed by preparing slurries of starting material HM30181 mesylate Type A in a variety of solvents and under a variety of conditions as described below. The resulting solids were analyzed by XRPD and identified for physical state. Results are summarized in Table 3 and Table 4.

Slurries of HM30181 mesylate Type A in MeOH generated HM30181 mesylate Type B after 1 week at temperatures of 4° C. to 50° C. (seeFIG.15). Slurries of HM30181 mesylate Type A in DCM at ambient temperature and acetonitrile at 4° C. to 50° C. generated HM30181 mesylate Type C (seeFIG.16). Slurries of HM30181 mesylate Type A in NMP at ambient temperature generated HM30181 mesylate Type D (seeFIG.17). Both HM30181 mesylate Type C and HM30181 mesylate Type D showed significant similarities to HM30181 mesylate Type A. Slurries of HM30181 mesylate Type A in DMA at ambient conditions generated HM30181 mesylate Type E (seeFIG.18). Slurries of HM30181 mesylate Type A in DMF generated HM30181 mesylate Type F at ambient temperature (seeFIG.19) and Type G at 50° C. (seeFIG.20).

Table 3 (continued in Table 4)APISolventCrystalExpMethodTemp.Solvent(mg)(mL)Type6007235-16-C10Slurry4°C.1,4-Dioxane25.90.2A6007235-16-B10Slurry50°C.1,4-Dioxane23.40.2A6007235-16-A10Slurryambient1,4-Dioxane210.2Atemperature6007235-16-A34Slurryambient1-Methyl-2-250.2Atemperaturepyrrolidinone/water(1:1)6007235-16-C9Slurry4°C.2-Methyl27.40.2Atetrahydrofuran6007235-16-B9Slurry50°C.2-Methyl24.50.2Atetrahydrofuran6007235-16-A9Slurryambient2-Methyl28.70.2Atemperaturetetrahydrofuran6007235-16-C4Slurry4°C.Acetone25.30.2A6007235-16-B4Slurry50°C.Acetone27.90.2A6007235-16-A4SlurryambientAcetone26.70.2Atemperature6007235-16-C13Slurry4°C.CHCl322.40.2A6007235-16-B13Slurry50°C.CHCl3290.1A6007235-16-A14SlurryambientCHCl325.60.2Atemperature6007235-16-C21Slurry4°C.Cumene22.10.2A6007235-16-B25Slurry50°C.Cumene26.30.2A6007235-16-A26SlurryambientCumene26.10.2Atemperature6007235-16-C17Slurry4°C.Cyclohexane24.20.2A6007235-16-B17Slurry50°C.Cyclohexane23.80.2A6007235-16-A18SlurryambientCyclohexane26.20.2Atemperature6007235-16-C24Slurry4°C.Cyclopentylmethyl20.80.2Aether6007235-16-B29Slurry50°C.Cyclopentylmethyl24.90.2Aether6007235-16-A30SlurryambientCyclopentylmethyl21.70.2Atemperatureether6007235-16-A22SlurryambientDMSO22.70.1Atemperature6007235-16-A31SlurryambientDMSO/water (1:1)25.10.2Atemperature6007235-16-C2Slurry4°C.Ethanol20.70.2A6007235-16-B2Slurry50°C.Ethanol26.90.2A6007235-16-A2SlurryambientEthanol21.70.2Atemperature6007235-16-C6Slurry4°C.Ethyl acetate26.20.2A6007235-16-B6Slurry50°C.Ethyl acetate25.50.2A6007235-16-A6SlurryambientEthyl acetate24.20.2Atemperature6007235-16-C22Slurry4°C.Ethyl formate24.80.2A6007235-16-B27Slurry50°C.Ethyl formate250.2A6007235-16-A28SlurryambientEthyl formate25.10.2Atemperature6007235-16-C23Slurry4°C.Isobutyl Acetate23.70.2A6007235-16-B28Slurry50°C.Isobutyl Acetate220.2A6007235-16-A29SlurryambientIsobutyl Acetate25.40.2Atemperature6007235-16-C3Slurry4°C.isopropanol26.80.2A6007235-16-B3Slurry50°C.isopropanol21.50.2A6007235-16-A3Slurryambientisopropanol23.60.2Atemperature6007235-16-C7Slurry4°C.isopropyl acetate25.90.2A6007235-16-B7Slurry50°C.isopropyl acetate21.90.2A6007235-16-A7Slurryambientisopropyl acetate28.80.2Atemperature6007235-16-C18Slurry4°C.methyl ethyl21.80.2Aketone6007235-16-B18Slurry50°C.methyl ethyl20.60.2Aketone6007235-16-A19Slurryambientmethyl ethyl27.10.2Atemperatureketone6007235-16-C5Slurry4°C.methyl isobutyl26.10.2Aketone6007235-16-B5Slurry50°C.methyl isobutyl270.2Aketone6007235-16-A5Slurryambientmethyl isobutyl27.90.2AtemperatureketoneTable 0 (continued from Table 3)APISolventCrystalExpMethodTemp.Solvent(mg)(mL)Type6007235-16-C11Slurry4°C.Methyl t-butyl ether27.10.2A6007235-16-B11Slurry50°C.Methyl t-butyl ether27.50.2A6007235-16-A11SlurryambientMethyl t-butyl ether26.80.2Atemperature6007235-16-A32SlurryambientN,N-26.90.2AtemperatureDimethylacetamide/water(1:1)6007235-16-A33SlurryambientN,N-270.2AtemperatureDimethylacetamide/water(1:1)6007235-16-C15Slurry4°C.n-Heptane26.80.2A6007235-16-B15Slurry50°C.n-Heptane22.70.2A6007235-16-A16Slurryambientn-Heptane22.80.2Atemperature6007235-16-C19Slurry4°C.n-Propanol22.30.2A6007235-16-B23Slurry50°C.n-Propanol21.80.2A6007235-16-A24Slurryambientn-Propanol23.30.2Atemperature6007235-16-C20Slurry4°C.n-Propyl acetate21.50.2A6007235-16-B24Slurry50°C.n-Propyl acetate27.40.2A6007235-16-A25Slurryambientn-Propyl acetate26.50.2Atemperature6007235-16-B19Slurry50°C.t-Butanol23.70.2A6007235-16-A20Slurryambientt-Butanol23.90.2Atemperature6007235-16-C8Slurry4°C.Tetrahydrofuran22.90.2A6007235-16-B8Slurry50°C.Tetrahydrofuran21.70.2A6007235-16-A8SlurryambientTetrahydrofuran20.90.2Atemperature6007235-16-C14Slurry4°C.Toluene24.20.2A6007235-16-B14Slurry50°C.Toluene25.70.2A6007235-16-A15SlurryambientToluene24.90.2Atemperature6007235-16-C16Slurry4°C.Water27.60.2A6007235-16-B16Slurry50°C.Water270.2A6007235-16-C28Slurry4°C.1-Methyl-2-23.90.2amorphous +pyrrolidinonee/waterA(1:1)6007235-16-C26Slurry4°C.N,N-26.40.2amorphous +Dimethylacetamide/waterA(1:1)6007235-16-C27Slurry4°C.N,N-240.2amorphous +Dimethylacetamide/waterA(1:1)6007235-16-A17SlurryambientWater220.2amorphous +temperatureA6007235-16-B20Slurry50°C.1-Methyl-2-pyrrolidinone23.70.1amorphous6007235-16-B21Slurry50°C.DMSO23.80.1amorphous6007235-16-C25Slurry4°C.DMSO/water (1:1)24.30.2amorphous6007235-16-A1SlurryambientMethanol24.90.2Btemperature6007235-16-B1Slurry50°C.Methanol270.1B6007235-16-C1Slurry4°C.Methanol23.20.2B6007235-16-A12SlurryambientAcetonitrile25.50.2Ctemperature6007235-16-A13SlurryambientDichloromethane21.90.2Ctemperature6007235-16-B12Slurry50°C.Acetonitrile22.70.1C6007235-16-C12Slurry4°C.Acetonitrile21.90.2C6007235-16-A21Slurryambient1-Methyl-2-pyrrolidinone24.60.2Dtemperature6007235-16-A23SlurryambientN,N-Dimethylacetamide25.60.1Etemperature6007235-16-A27SlurryambientN,N-Dimethylacetamide26.20.1Ftemperature6007235-16-B26Slurry50°C.N,N-Dimethylacetamide27.30.1G

Generation and/or screening of HM30181 mesylate polymorphs was also performed by preparing HM30181 mesylate Type A starting material for liquid and solid vapor diffusion as described below. Resulting solids were analyzed by XRPD and identified for physical state. Results are summarized in Table 5, Table 6, and Table 7. Liquid vapor diffusion of MTBE into NMP solution yielded HM30181 mesylate Type D (seeFIG.21). Liquid vapor diffusion of MEK into NMP or DMA solution yielded HM30181 mesylate Type D (seeFIG.21). Some loss of crystallinity was noted on air-dried HM30181 mesylate Type D, suggesting possible solvate. Liquid vapor diffusion of 2-MeTHF into DMA or DMF solution yielded HM30181 mesylate Type E (seeFIG.22). Liquid vapor diffusion of MTBE into DMA solution yielded HM30181 mesylate Type E, with a few additional diffraction peaks, which Inventors believe are attributable to HM30181 mesylate Type I (seeFIGS.4to8). Liquid vapor diffusion of ACN into DMSO solution yielded HM30181 mesylate Type H (seeFIG.23). Liquid vapor diffusion of Acetone, Ethyl Acetate, or isopropyl acetate into DMA solution yielded HM30181 mesylate Type I (seeFIG.24). Air-drying of HM30181 mesylate Type I yielded HM30181 mesylate Type J (seeFIG.24). Liquid vapor diffusion of isopropyl acetate into DMSO solution followed by air drying also yielded HM30181 mesylate Type J.

TABLE 5APISolventCrystalExpTemperatureSolvent(mg)(mL)Type6007235-17-A1ambientmethanol25.20.1Atemperature6007235-17-A2ambientethanol22.10.1Atemperature6007235-17-A3ambientisopropanol21.40.1Atemperature6007235-17-A4ambientacetone24.80.1Atemperature6007235-17-A5ambientmethyl isobutyl ketone25.80.1Atemperature6007235-17-A6ambientethyl acetate22.90.1Atemperature6007235-17-A7ambientisopropyl acetate23.80.1Atemperature6007235-17-A8ambienttetrahydrofuran24.50.1Atemperature6007235-17-A9ambient2-methyl tetrahydrofuran24.90.1Atemperature6007235-17-A10ambientmethyl t-butyl ether27.10.1Atemperature6007235-17-A11ambientacetonitrile27.70.1Atemperature6007235-17-A12ambientdichloromethane24.90.1Atemperature6007235-17-A13ambientCHCl3290.1Atemperature6007235-17-A14ambientmethyl ethyl ketone23.40.1Atemperature6007235-17-A15ambientt-butanol28.20.1Atemperature6007235-17-A16ambientn-propanol25.40.1Atemperature6007235-17-A17ambientethyl formate27.30.1Atemperature6007235-17-A18ambientcyclopentylmethyl ether25.10.1Atemperature6007235-17-A1925° C.25° C./60% RH22.90.1A6007235-17-A2040° C.40° C./75% RH23.50.1A6007235-17-A2140° C.100% RH(water)22.70.1A

Table 6 (continued on Table 7)Anti-APISolventsolventExpTemp.Solvent(mg)(mL)Anti-solvent(mL)Crystal Type6007235-19-A1ambientmethanol2216ethyl formate0.5No solidstemperature6007235-19-A2ambientmethanol2316dichloromethane0.5No solidstemperature6007235-19-B1ambientDMSO250.3ethanol2No solidstemperature6007235-19-B2ambientDMSO250.3isopropanol2No solidstemperature6007235-19-B3ambientDMSO250.3acetone2Amorphoustemperature6007235-19-B4ambientDMSO250.3MIBK2Amorphoustemperature6007235-19-B5ambientDMSO250.3ethyl acetate2Amorphoustemperature6007235-19-B7ambientDMSO250.3tetrahydrofuran2Amorphoustemperature6007235-19-B8ambientDMSO250.32-MeTHF2Amorphoustemperature6007235-19-B9ambientDMSO250.3methyl t-butyl2Amorphoustemperatureether6007235-19-B11ambientDMSO250.3dichloromethane2Amorphoustemperatureand A6007235-19-B12ambientDMSO250.3methyl ethyl2Amorphoustemperatureketone6007235-19-B13ambientDMSO250.3t-butanol2No solidstemperature6007235-19-C1ambientNMP303ethanol3No solidstemperature6007235-19-C2ambientNMP303isopropanol3No solidstemperature6007235-19-C3ambientNMP303acetone3No solidstemperature6007235-19-C4ambientNMP303MIBK3No solidstemperature6007235-19-C5ambientNMP303ethyl acetate3Amorphoustemperatureand A6007235-19-C6ambientNMP303isopropyl acetate3No solidstemperature6007235-19-C7ambientNMP303tetrahydrofuran3No solidstemperature6007235-19-C8ambientNMP3032-MeTHF3No solidstemperature6007235-19-C10ambientNMP303acetonitrile3No solidstemperature6007235-19-C11ambientNMP303dichloromethane3No solidstemperature6007235-19-C13ambientNMP303t-butanol3No solidstemperature6007235-19-D1ambientDMF303ethanol3No solidstemperature6007235-19-D2ambientDMF303isopropanol3Amorphoustemperature6007235-19-D4ambientDMF303MIBK3No solidstemperature6007235-19-D5ambientDMF303ethyl acetate3Amorphoustemperature6007235-19-D6ambientDMF303isopropyl acetate3Amorphoustemperature6007235-19-D7ambientDMF303tetrahydrofuran3No solidstemperature6007235-19-D9ambientDMF303methyl t-butyl3Amorphoustemperatureether6007235-19-D10ambientDMF303acetonitrile3No solidstemperature6007235-19-D11ambientDMF303dichloromethane3No solidstemperature6007235-19-D12ambientDMF303methyl ethyl3Amorphoustemperatureketone6007235-19-D13ambientDMF303t-butanol3No solidstemperature6007235-19-E1ambientDMA204ethanol8No solidstemperature6007235-19-E2ambientDMA204isopropanol8No solidstemperature6007235-19-E4ambientDMA204MIBK8No solidstemperature6007235-19-E7ambientDMA204tetrahydrofuran8No solidstemperature6007235-19-E10ambientDMA204acetonitrile8No solidstemperature6007235-19-E11ambientDMA204dichloromethane8No solidstemperature6007235-19-E13ambientDMA204t-butanol8No solidstemperatureTable 7 (continued from Table 6)Anti-XRPDAPISolventsolvent(wetXRPDExp.Temp.(mg)Solvent(mL)Anti-solvent(mL)cake)(air-dry)6007235-19-C12ambient30NMP3MEK3Type D—temperature6007235-19-E12ambient20DMA4MEK8Type DType D andtemperatureamorphous6007235-19-C9ambient30NMP3MTBE3Type D—temperature6007235-19-D8ambient30DMF32-MeTHF3Type EEtemperature6007235-19-E8ambient20DMA42-MeTHF8Type EEtemperature6007235-19-E9ambient20DMA4MTBE8Type EEtemperatureand I6007235-19-B10ambient25DMSO0.3ACN2Type H—temperature6007235-19-E3ambient20DMA4Acetone8Type IType Jtemperature6007235-19-E5ambient20DMA4EtOAc8Type IType Jtemperature6007235-19-E6ambient20DMA4iPrOAc8Type IType Jtemperature6007235-19-B6ambient25DMSO0.3iPrOAc3—Type Jtemperature

Generation and/or screening of HM30181 mesylate polymorphs by cooling was carried out by treating HM30181 mesylate Type A starting material using gradual or rapid (i.e. crash) cooling as described below. The resulting solids were analyzed by XRPD and identified for physical state. Results are summarized in Table 8. Cooling experiments in acetonitrile and DCM yielded HM30181 mesylate Type C; no significant change was noted on air-drying (seeFIG.25).

TABLE 8Solvent/Solvent/Anti-Anti-APIsolventXRPDXRPDExp.Temp.Tempsolvent(mg)(mL)(wet cake)(air-dry)6007235-17-C6Slow Cooling50° C.-->4° C.NMP20.94No solids6007235-17-C14Crash Cooling50° C.-->−20° C.NMP26.24Amorphous6007235-17-C2Slow Cooling50° C.-->4° C.CHCl325.44Amorphousand A6007235-17-C10Crash Cooling50° C.-->−20° C.CHCl325.44No solids6007235-17-C13Crash Cooling50° C.-->−20° C.DCM22.110No solids6007235-17-C3Slow Cooling50° C.-->4° C.EtOH520No solids6007235-17-C11Crash Cooling50° C.-->−20° C.EtOH520No solids6007235-17-C9Crash Cooling50° C.-->−20° C.MeOH22.910No solids6007235-17-C1Slow Cooling50° C.-->4° C.MeOH24.720No solids6007235-17-C7Slow Cooling50° C.-->4° C.DMA254Amorphousand A6007235-17-C15Crash Cooling50° C.-->−20° C.DMA26.94Amorphous6007235-17-C8Slow Cooling50° C.-->4° C.DMF25.14No solids6007235-17-C16Crash Cooling50° C.-->−20° C.DMF25.14Amorphous6007235-17-C4Slow Cooling50° C.-->4° C.ACN12.610Type CType C6007235-17-C5Slow Cooling50° C.-->4° C.DCM26.810Type C +Type Cpeaks6007235-17-C12Crash cooling50° C.-->−20° C.ACN12.610Type CType C

HM30181 mesylate was also subjected to evaporation methods by treating HM30181 mesylate Type A starting material as described below. The resulting solids were analyzed by XRPD and identified for physical state. Results are shown in Table 9.

TABLE 9APISolventExpTemp.Solvent(mg)(mL)Crystal Type6007235-17-B6ambient temperaturemethanol1210Amorphous6007235-17-B1ambient temperaturemethanol1210Amorphous6007235-17-B850° C.ethanol520Amorphous and A6007235-17-B350° C.ethanol520Amorphous6007235-17-B550° C.dichloromethane21.220Amorphous6007235-17-B1050° C.dichloromethane2820Amorphous6007235-17-B7ambient temperatureCHCl327.47Amorphous6007235-17-B2ambient temperatureCHCl3267Amorphous6007235-17-B950° C.acetonitrile10.540Amorphous6007235-17-B450° C.acetonitrile10.540Amorphous

Generation and/or screening of HM30181 mesylate polymorphs by treatment with anti-solvents was performed by treating HM30181 mesylate Type A starting material as described below. The resulting solids were analyzed by XRPD and identified for physical state. Results are summarized in Table 10, Table 11, Table 12, and Table 13. Anti-solvent studies in DMSO yielded HM30181 mesylate Type C (seeFIG.26). Anti-solvent studies in N,N-dimethylacetamide with methyl t-butyl ether yielded a HM30181 mesylate Type F polymorphism (seeFIG.27). Anti-solvent addition and reverse anti-solvent addition in DMSO/EtOAc, DMA/MIBK, DMA/toluene and NMP/t-BuOH yielded primarily amorphous content and a polymorphism with some similarity to HM30181 mesylate Type J (seeFIG.28). Other anti-solvent studies in DMSO and DMF yielded a HM30181 mesylate Type K polymorphism (or possibly a mixture of types, seeFIG.29). Anti-solvent experiments in DMF/n-propanol and DMA/isopropanol yielded a HM30181 mesylate Type L polymorphism (seeFIG.30). Anti-solvent addition DMF/Toluene and DMA/t-BuOH were mostly amorphous, but also generated HM30181 mesylate Type M (seeFIG.31).

Table 10 (continued on Table 11)Anti-Anti-solventEvapNBSolventsolvent(mL)Precipitation?solids?Identification6004273-06-A1DMSOEtOH20NYType K6004273-06-A2DMSOIPA10Yinsufficientsolids/gel6004273-06-A3DMSOAcetone10YType K6004273-06-A4DMSOMIBK10YType K6004273-06-A5DMSOEtOAc10Yinsufficientsolids/gel6004273-06-A6DMSOiPrOAc10Yinsufficientsolids/gel6004273-06-A7DMSOTHF10YSimilar toType K6004273-06-A8DMSO2-MeTHF10YAmorphous6004273-06-A9DMSO1,4-Dioxane20Yinsufficientsolids/gel6004273-06-A10DMSOMTBE20YAmorphous6004273-06-A11DMSOACN20NYType C6004273-06-A12DMSODCM20NYSimilar toType C6004273-06-A13DMSOCHCl320NYType K6004273-06-A14DMSOToluene10YAmorphous6004273-06-A15DMSOWater20NNinsufficientsolids/gel6004273-06-A16DMSOMEK10Yinsufficientsolids/gel6004273-06-A17DMSOt-Butanol10YType K6004273-06-A18DMSOn-Propanol10YType K6004273-06-A19DMSOn-Propyl10YType Kacetate6004273-06-A20CHCl3Ethanol10YType A6004273-06-A21CHCl3IPA10YType A6004273-06-A22CHCl3Acetone10YType A6004273-06-A23CHCl3MIBK10YType A6004273-06-A24CHCl3EtOAc10YType A6004273-06-A25CHCl3iPrOAc10YType A6004273-06-A26CHCl3THF10YType A6004273-06-A27CHCl32-MeTHF10YType A6004273-06-A28CHCl31,4-Dioxane10Yinsufficientsolids/gel6004273-06-A29CHCl3MTBE10YType A6004273-06-A30CHCl3ACN10YType A6004273-06-A31CHCl3DCM10YType A6004273-06-A32CHCl3Toluene10YType A6004273-06-A33CHCl3n-Heptane10YType ATable 11 (continued from Table 10 and on Table 12)Anti-solventEvapNBSolventAnti-solvent(mL)Precipitation?solids?Identification6004273-06-A34CHCl3MeOAc10YType A6004273-06-A35CHCl3Cyclohexane10YType A6004273-06-A36CHCl3MEK10YType A6004273-06-A37CHCl3t-Butanol10YType A6004273-06-A38CHCl3n-Propanol10YType A6004273-06-A39CHCl3n-Propyl10YType Aacetate6004273-06-A40CHCl3Ethyl formate10YType A6004273-06-A41CHCl3iBuOAc10YType A6004273-06-A42CHCl3CPME10YType A6004273-06-A43DMFEtOH20NYAmorphous6004273-06-A44DMFIPA10YType K6004273-06-A45DMFAcetone20NYinsufficientsolids/gel6004273-06-A46DMFMIBK10YAmorphous6004273-06-A47DMFEtOAc10YType A6004273-06-A48DMFiPrOAc10Yinsufficientsolids/gel6004273-06-A49DMFTHF20NYinsufficientsolids/gel6004273-06-A50DMF2-MeTHF10Yinsufficientsolids/gel6004273-06-A51DMF1,4-Dioxane20NYinsufficientsolids/gel6004273-06-A52DMFMTBE10YType A6004273-06-A53DMFACN20NYinsufficientsolids/gel6004273-06-A54DMFDCM20NYinsufficientsolids/gel6004273-06-A55DMFCHCl320NYinsufficientsolids/gel6004273-06-A56DMFToluene10YType M6004273-06-A57DMFWater20NYinsufficientsolids/gel6004273-06-A58DMFMEK20NYinsufficientsolids/gel6004273-06-A59DMFt-Butanol10Yinsufficientsolids/gel6004273-06-A60DMFn-Propanol20NYType L +amorphous6004273-06-A61DMFn-Propyl10YType Aacetate6004273-06-A62NMPEtOH20NNinsufficientsolids/gel6004273-06-A63NMPIPA10Yinsufficientsolids/gel6004273-06-A64NMPAcetone20NNinsufficientsolids/gel6004273-06-A65NMPMIBK10Yinsufficientsolids/gel6004273-06-A66NMPEtOAc10Yinsufficientsolids/gelTable 12 (continued from Table 11)Anti-solventEvapNBSolventAnti-solvent(mL)Precipitation?solids?Identification6004273-06-A67NMPiPrOAc10Yinsufficientsolids/gel6004273-06-A68NMPTHF20NNinsufficientsolids/gel6004273-06-A69NMP2-MeTHF10Yinsufficientsolids/gel6004273-06-A70NMP1,4-Dioxane20NNType A6004273-06-A71NMPMTBE10Yinsufficientsolids/gel6004273-06-A72NMPACN20NNinsufficientsolids/gel6004273-06-A73NMPDCM20NNinsufficientsolids/gel6004273-06-A74NMPCHCl320NNinsufficientsolids/gel6004273-06-A75NMPToluene10Yinsufficientsolids/gel6004273-06-A76NMPWater20NNinsufficientsolids/gel6004273-06-A77NMPMEK20NYAmorphous6004273-06-A78NMPt-BuOH20hazyYAmorphous +Type J6004273-06-A79NMPn-Propanol20NNinsufficientsolids/gel6004273-06-A80NMPn-Propyl10Yinsufficientacetatesolids/gel6004273-06-A81DMAEtOH20NNinsufficientsolids/gel6004273-06-A82DMAIPA20hazyYType L +amorphous6004273-06-A83DMAAcetone20NYinsufficientsolids/gel6004273-06-A84DMAMIBK10YAmorphous +Type J6004273-06-A85DMAEtOAc10Yinsufficientsolids/gel6004273-06-A86DMAiPrOAc10Yinsufficientsolids/gel6004273-06-A87DMATHF20NNinsufficientsolids/gel6004273-06-A88DMA2-MeTHF10Yinsufficientsolids/gel6004273-06-A89DMA1,4-Dioxane20NNinsufficientsolids/gel6004273-06-A90DMAMTBE10YAmorphous +Type F6004273-06-A91DMAACN20NNinsufficientsolids/gel6004273-06-A92DMADCM20NNinsufficientsolids/gel6004273-06-A93DMACHCl320NNinsufficientsolids/gel6004273-06-A94DMAToluene10YAmorphous +Type J6004273-06-A95DMAWater20NYinsufficientsolids/gel6004273-06-A96DMAMEK20NNinsufficientsolids/gel6004273-06-A97DMAt-Butanol20NYType M6004273-06-A98DMAn-Propanol20NYinsufficientsolids/gel6004273-06-A99DMAn-Propyl10Yinsufficientacetatesolids/gel

TABLE 13Anti-solventEvapNBSolventAnti-solvent(mL)Precipitation?solids?Identification6004273-06-A100NMP1,4-Dioxane20Ninsufficientsolids/gel6004273-06-A101NMPMIBK10Yinsufficientsolids/gel6004273-06-A102CHCl3EtOH10YType A6004273-06-A103CHCl3MTBE10YType A6004273-06-A104CHCl3n-Heptane10YType A6004273-06-A105DMSOEtOAc10Yinsufficientsolids/gel6004273-06-A106DMSOToluene10YAmorphous6004273-06-A107DMSOWater20Ninsufficientsolids/gel6004273-06-A108DMAAcetone20Ninsufficientsolids/gel6004273-06-A109DMAiPrOAc10Yinsufficientsolids/gel6004273-06-A110DMFIPA10YAmorphous +Type K6004273-06-A111DMFTHF20Ninsufficientsolids/gel

Large scale studies were performed with HM30181 mesylate Type A starting material on a 200 mg scale as described below. Results are summarized in Table 14. Solids were isolated by vacuum filtration. The wet cakes from filtration from DMA, DMF, and NMP slurries were washed with 1 mL to 2 mL methanol to remove solvents. Solids were then vacuum dried at 80° C. overnight.

At large scale, a slurry of HM30181 mesylate Type A starting material in methanol at ambient conditions generated a mixture of HM30181 mesylate Type B and HM30181 mesylate Type A after 9 days (seeFIG.32). After 14 days, the slurry in methanol generated a HM30181 mesylate Type N polymorph. HM30181 mesylate Type N showed some loss of crystallinity after vacuum-drying, suggesting a methanol solvate (seeFIG.32). A slurry of HM30181 mesylate Type A starting material in acetonitrile at ambient conditions generated HM30181 mesylate Type C after 14 days; no loss of crystallinity was detected after vacuum-drying (seeFIG.33). A scaled-up slurry of HM30181 mesylate Type A starting material in NMP provided a primarily amorphous material (seeFIG.34). A scaled-up slurry of HM30181 mesylate Type A starting material in DMA yielded HM30181 mesylate Type E after 6 days (seeFIG.35). HM30181 mesylate Type E showed no loss of crystallinity after vacuum drying. A scaled-up slurry of HM30181 mesylate Type A starting material in DMF at 50° C. yielded HM30181 mesylate Type F rather than the expected HM30181 mesylate Type G after 9 days (seeFIG.36). Some change of pattern was noted after vacuum-drying. A scaled-up slurry of HM30181 mesylate Type A starting material in DMF at ambient temperature initially showed no change from Type A (seeFIG.37). This slurry was heated in an attempt to generate HM30181 mesylate Type G. A scaled-up slurry of HM30181 mesylate Type A starting material in DMF at 100° C. yielded HM30181 mesylate Type F after 2 days (seeFIG.37). A scaled-up slurry of HM30181 mesylate Type A starting material in DMF at 150° C. yielded a previously unobserved XRPD pattern after 5 days (seeFIG.37). 1H-NMR results in DMSO-d6suggested that the DMF slurry at 150° C. resulted in degradation, as the 1H-NMR spectrum did not match either the starting material or freebase (seeFIG.38).

By 1H-NMR, no disproportionation or degradation was detected in HM30181 mesylate Types C, E, F and N polymorphs (seeFIG.39). By 1H-NMR, HM30181 mesylate Type E contained DMA, HM30181 mesylate Type F contains DMF, and HM30181 mesylate Type N contains MeOH.

TABLE 14Anti-SolventConc.Anti-solventDesiredNBMethodSolvent(mL)(mg/mL)solvent(mL)TypeObservations6004273-10-BSlurry atMeOH0.2480BType Nambienttemperature6004273-10-CSlurry atMeCN0.2545CType Cambienttemperature6004273-10-DSlurry atNMP0.2550DAmorphousambienttemperature6004273-10-ESlurry atDMA0.2576EType Eambienttemperature6004273-10-FSlurry atDMF0.2595FType Aambienttemperature6004273-10-Slurry atDMF0.2595GType FF-100C100° C.(for 2 days)6004273-10-Slurry atDMF0.2595GDecompositionF-150C150° C.(for 5 d)6004273-10-GSlurry atDMF0.2576GType F50° C.6004273-10-HLiquidDMSO560MeCN3HNo precipitation,vaporevaporated- nosorptionsolids due toDMSO6004273-10-JLiquidDMSO1060Acetone3JNo precipitation,vaporevaporated- nosorptionsolids due toDMSO6004273-10-KAnti-DMSO1060MIBK200KAmorphoussolventaddition6004273-10-LAnti-DMF205n-propanol200LAmorphoussolventaddition6004273-10-MAnti-DMA205t-BuOH200MAmorphoussolventaddition6004273-10-M1Anti-DMF205Toluene300MAmorphoussolventaddition

Characteristics of HM30181 mesylate salt polymorphisms as characterized by XRPD, TGA, DSC, and DVS are summarized below:HM30181 mesylate Type A represents a prior art preparation of HM30181 mesylate that can be used as a starting material in generation of novel polymorphs of this compound.Crystalline HM30181 mesylate Type B was obtained through slurrying in methanol at 4° C. to 50° C. and is distinct by XRPD (seeFIG.40). By DSC, HM30181 mesylate Type B displayed an endotherm at 159.92° C. (seeFIG.41). By TGA, HM30181 mesylate Type B showed 1.865% weight loss before 170° C., followed by possible disassociation and decomposition (seeFIG.41).Crystalline HM30181 mesylate Type C polymorph was obtained through slurrying in acetonitrile at ambient temperature and shows characteristic results on XRPD. No change in crystalline pattern was noted after vacuum drying overnight (seeFIG.42). By DSC, HM30181 mesylate Type C displays an endotherm at 159.60° C. (seeFIG.43). By TGA, HM30181 mesylate Type C showed 2.987% weight loss before 170° C., followed by a disassociation and decomposition (seeFIG.43). By 1H-NMR, HM30181 mesylate Type C is confirmed to contain only water and no acetonitrile, (˜2.07 ppm) suggesting a monohydrate (seeFIG.44).Crystalline HM30181 mesylate Type D was obtained through slurrying in N-methyl pyrrolidone at ambient temperature and shows characteristic results by XRPD. No change in crystalline pattern was noted after air drying overnight (seeFIG.45). By DSC/TGA, HM30181 mesylate Type D displayed an endotherm at 66.97° C. and a 14.71% weight loss before 150° C., suggesting significant residual solvent content (seeFIG.46).Crystalline HM30181 mesylate Type E was obtained through slurrying in N,N-dimethylacetamide at ambient temperature and shows characteristic results by XRPD. No change in crystalline pattern was noted after vacuum drying overnight at 80° C. (seeFIG.47). By DSC/TGA, HM30181 mesylate Type E displayed an endotherm at 154.42° C. and a 5.247% weight loss before 168° C., suggesting HM30181 mesylate Type E was a solvate (seeFIG.48). By1H-NMR, HM30181 mesylate Type E was confirmed to contain DMA (seeFIG.49).Crystalline HM30181 mesylate Type F was obtained through slurrying in dimethylformamide at 50° C. and shows characteristic results by XRPD (seeFIG.50). A significant reduction in crystallinity was noted after vacuum drying overnight at 80° C., suggesting HM30181 mesylate Type F is a metastable solvate. By DSC/TGA, HM30181 mesylate Type F displayed an endotherm at 148.41° C. and a 5.123% weight loss before 180° C., consistent with loss of DMF (seeFIG.51). By1H-NMR, HM30181 mesylate Type F was confirmed to contain DMF (seeFIG.52).Crystalline HM30181 mesylate Type G was obtained through slurrying in dimethylformamide at 50° C. and shows characteristic results by XRPD. No reduction in crystallinity was noted after air drying overnight (seeFIG.53). By DSC/TGA, HM30181 mesylate Type G displayed an endotherm at 69.02° C. and at 233.29° C. and a 1.451% weight loss before 150° C., which was consistent with loss of DMF (seeFIG.54).Crystalline HM30181 mesylate Type H was obtained through liquid vapor diffusion of acetonitrile into a DMSO stock of HM30181 mesylate and shows characteristic results by XRPD (seeFIG.55). By DSC/TGA, HM30181 mesylate Type H displayed an endotherm at 126.52° C. and a 7.444% weight loss before 200° C., potentially from residual ACN and DMSO or from a solvate (seeFIG.56).Crystalline HM30181 mesylate Type I was obtained through liquid vapor diffusion of acetone, ethyl acetate, or isopropyl acetate into a DMA stock of HM30181 mesylate and shows characteristic results by XRPD (seeFIG.57). Air-drying of HM30181 mesylate Type I yielded HM30181 mesylate Type J.HM30181 mesylate Type J was obtained through liquid vapor diffusion of acetone, ethyl acetate, or isopropyl acetate into a DMA stock or isopropyl acetate into a DMSO stock of HM30181 mesylate followed by air-drying. By XRPD, HM30181 mesylate Type J is crystalline (seeFIG.58). By TGA, HM30181 mesylate Type J displayed a 2.887% weight loss before 150° C., suggesting a solvate or hydrate (seeFIG.59).HM30181 mesylate type K was obtained through anti-solvent addition using DMF/IPA and multiple DMSO systems (ethanol, acetone, MIBK, THF, chloroform, t-butanol, n-propyl acetate, and n-propanol). By XRPD, HM30181 mesylate Type K is partially crystalline (seeFIG.60).HM30181 mesylate Type L was obtained through anti-solvent addition in DMF/n-propanol and DMA/isopropanol systems. By XRPD, HM30181 mesylate Type L is partially crystalline (seeFIG.61).HM30181 mesylate type M was obtained through anti-solvent addition in DMF/toluene and DMA/t-butanol systems. By XRPD, HM30181 mesylate Type M is partially crystalline (seeFIG.62).

Crystalline HM30181 mesylate Type N was obtained after 14-days treatment of HM30181 mesylate Type A starting material as a slurry in methanol at ambient temperature (seeFIG.63). HM30181 mesylate Type N showed some loss of crystallinity after vacuum-drying, suggesting a methanol solvate. By DSC, HM30181 mesylate Type N displayed endotherms at 159.28° C. and 188.47° C. with a 2.128% weight loss before 180° C., followed by possible disassociation and decomposition (seeFIG.64). 1H-NMR confirmed the presence of methanol (seeFIG.65).

HM30181 mesylate Type C and E forms were further analyzed to determine unit cell dimensions. Unit cell parameters for the Type C polymorph of HM30181 mesylate were calculated using cumulative XRPD spectra, peak identifications for which are shown in Table 15. Notably distinct peaks for HM30181 mesylate Type C are shown in bold and italicized in Table 15. Estimated values of unit cell parameters derived from the Type C polymorph of HM30181 mesylate are shown in Table 16 and are consistent with triclinic P unit cells.

TABLE 15FWHMRel.Pos.HeightLeftd-spacingInt.[°2Th.][cts][°2Th.][Å][%]3.534.60.0525.51.85.195.50.11517.55.05.630.20.0815.81.66.4419.60.0813.722.28.01247.30.1211.065.99.937.70.208.92.011.7305.90.207.616.212.1322.70.107.317.112.81892.00.106.9100.013.9205.80.136.410.915.0609.30.135.932.215.5113.80.825.76.016.1253.90.155.513.417.871.20.465.03.819.2439.50.204.623.219.7590.30.204.531.220.0362.70.204.419.221.260.70.104.23.222.120.00.104.01.022.9309.00.133.916.323.499.70.183.85.324.537.20.203.62.025.163.60.313.63.426.11066.20.313.456.426.9130.00.203.36.928.1144.20.233.27.629.415.40.313.00.832.419.00.202.81.034.325.90.052.61.435.5013.20.412.50.736.516.10.152.50.937.312.00.082.40.6

TABLE 16Reflection ConditionsCrystal SystemTriclinicBravais TypePrimitive (P)Space GroupInstrument SettingsGoniometer Radius (mm)240.00Unit Cella (Å)7.34 (2)b (Å)14.6 (1)c (Å)17.5 (8)Alpha (°)51.8 (6)Beta (°)62.3 (2)Gamma (°)90.4 (3)Volume (Å3)1179.70Refinement ResultsNo. Unindexed Lines0No. Indexed Lines17Total No.5110Calculated LinesChi Square3.777537E−0006Snyder's FOM2.0791

Characteristic XRPD peak values for the Type E polymorph are provided in Table 17, where notably distinct peaks are indicated by bolded and italicized numerals. It should be appreciated that these are distinct and different from those of polymorph Type C, indicating that the Type C and Type E polymorphs are distinct and different from one another and that both Type C and Type E polymorphs are distinct and different from the prior art Type A polymorph of HM30181 mesylate.

TABLE 17FWHMRel.Pos.HeightLeftd-spacingInt.[°2Th.][cts][°2Th.][Å][%]4.2458.00.1521.243.75.24.10.0517.00.46.016.00.03814.71.56.213.50.05114.31.36.322.10.03814.02.18.21047.30.1510.8100.09.035.80.0389.93.49.129.70.159.72.89.913.90.059.01.310.4344.20.138.532.910.7430.10.158.341.111.6136.80.207.613.112.3114.70.187.211.012.9170.30.156.916.313.630.80.156.52.914.7685.30.266.065.415.4227.70.135.721.815.9801.80.235.676.616.346.50.155.44.416.8827.70.135.379.017.7281.70.205.026.918.3200.10.234.919.118.9298.00.204.728.419.3164.20.134.615.719.6106.50.134.510.220.0103.10.154.49.820.4153.20.134.314.621.0842.30.224.280.421.3320.90.134.230.621.6243.70.104.123.322.0111.10.204.010.622.4128.70.154.012.322.7418.20.263.939.923.8272.60.263.726.024.1116.90.103.711.224.6174.50.203.616.724.9214.90.183.620.526.2994.90.153.495.026.6965.70.153.392.227.7343.30.203.232.828.394.30.183.29.028.820.20.153.11.929.362.10.153.05.930.745.90.202.94.432.1117.50.182.811.233.340.20.152.73.833.942.70.312.64.134.733.40.262.63.235.722.60.202.52.237.029.80.152.42.8
Unit cell parameters for the Type E polymorph of HM30181 mesylate were calculated using cumulative XRPD spectra. Estimated values of unit cell parameters derived from the Type E polymorph of HM30181 mesylate are shown in Table 18 and are consistent with triclinic P unit cells.

TABLE 18Reflection ConditionsCrystal SystemTriclinicBravais TypePrimitive (P)Space GroupInstrument SettingsGoniometer Radius (mm)240.00Unit Cella (Å)8.2(2)b (Å)9.8(2)c (Å)23.7(4)Alpha (°)75.2(2)Beta (°)78.66(3)Gamma (°)111.69(3)Volume (Å3)1618.45Refinement ResultsNo. Unindexed Lines0No. Indexed Lines32Total No.6088Calculated LinesChi Square8.802423E−0007Snyder's FOM5.7186

As noted above, HM30181 is an inhibitor of P-glycoprotein, an efflux transport protein that is effective at removing a wide range of therapeutic from cells and forms an important part of the blood brain barrier. While this function is essentially protective, it can adversely impact the use of therapeutic drugs that P-glycoprotein substrates. Examples of drugs that are transported by P-glycoprotein include, but are not limited to, antineoplastic drugs (e.g., docetaxel, etoposide, vincristine), calcium channel blockers (e.g., amlodipine), calcineurin inhibitors (e.g., cyclosporin, tacrolimus), digoxin, macrolide antibiotics (e.g., clarithromycin), and protease inhibitors. Accordingly, HM30181 mesylate can be used to alter the pharmacokinetics of therapeutic drug substrates of P-glycoprotein by reducing efflux of such drugs from the cells of an individual undergoing treatment.

Conventional process for the production of HM30181 provide the Type A polymorph. Inventors have produced and identified a number of other forms of this compound, including Type B, Type C, Type D, Type E, Type F, Type G, Type H, Type I, Type J, Type K, Type L, Type M, and Type N polymorphs of HM30181. As shown above, these are different and distinct from the prior art Type A polymorph and from each other. Inventors believe that these new polymorphs of HM30181 can provide different stabilities and/or pharmacokinetics (e.g., rate of absorption, etc.) than those provided by the prior art Type A polymorph.

Accordingly, another embodiment of the inventive concept is the application of one or more of a Type B, Type C, Type D, Type E, Type F, Type G, Type H, Type I, Type J, Type K, Type L, Type M, and/or Type N polymorph of HM30181 to inhibit P-glycoprotein, and in turn alter the pharmacokinetics of a drug that is a substrate of P-glycoprotein. In some of such embodiments the drug can be a chemotherapeutic drug used in the treatment of cancer.

In such embodiments one or more of a Type B, Type C, Type D, Type E, Type F, Type G, Type H, Type I, Type J, Type K, Type L, Type M, and/or Type N polymorph of HM30181 can be administered in concert with a drug that is a P-glycoprotein substrate to an individual that is in need of treatment for a disease or condition that is responsive to such a drug. In some embodiments a Type B, Type C, Type D, Type E, Type F, Type G, Type H, Type I, Type J, Type K, Type L, Type M, and/or Type N polymorph of HM30181 can be provided as a separate formulation. Alternatively, one or more of a Type B, Type C, Type D, Type E, Type F, Type G, Type H, Type I, Type J, Type K, Type L, Type M, and/or Type N polymorph 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 drug that is a P-glycoprotein substrate is a chemotherapeutic drug used to treat cancer.

Methods

As noted above, polymorphs of HM30181 mesylate were provided by treatment of a conventional HM30181 mesylate Type A preparation with a variety of solvents, and using a range of techniques. For solubility studies of HM30181 mesylate Type A in a variety of solvents a sample (˜2 mg) of the solid was transferred into a 4-mL glass vial. Solvent was added to the vial in a stepwise fashion, 50 μL per step until 100 μL total volume followed by 100 μL per step until concentration was less than 1.0 mg/mL. Samples were mixed thoroughly after each addition by sonication for 2 minutes and vortexing for 1 minute. Volumes of solvent (V1 and V2) were recorded and used to estimate solubility. Solvents used are summarized below in Table 19.

TABLE 19AbbreviationSolventAbbreviationSolventMeOHMethanolTHFTetrahydrofuranEtOHEthanol2-MeTHF2-MethyltetrahydrofuranIPAIsopropylDMFDimethyl formamidealcoholACNAcetonitrileDMSODimethyl sulfoxideMIBKMethyl isobutylCHCl3ChloroformketoneEtOAcEthyl acetateDCMDichloromethaneiPrOAcIsopropylDMAcDimethylacetamideacetateMTBEMethyl tert-butylt-BuOHt-Butanolether

Screening of polymorphisms of HM30181 mesylate can include preparation of a slurry. Typically, a slurry was prepared by suspending 5 mg to 20 mg of sample in 0.1 mL to 0.5 mL solvent in a 1.5 mL or 3.0 mL glass vial. The suspension a was stirred at target temperature (e.g. 4° C., ambient temperature, 50° C.) at 200 rpm. Solids for X-ray powder diffraction (XRPD) analysis were separated by centrifuging at 14,000 rpm for 5 minutes at ambient temperature. If no solid or gel is obtained, the slurry can be move to a fume hood for evaporation of the solvent.

In some embodiments anti-solvent addition was used. In this method a concentrated stock of compound in solvent is provided and an anti-solvent quickly added to the concentrated solution while stirring to induce precipitation. Solids can be isolated for XRPD analysis using filtration or centrifugation.

In some embodiments reverse anti-solvent addition was used. In this method a concentrated stock of compound in solvent is provided and quickly added to an anti-solvent with stirring to induce precipitation. Solids can be isolated for XRPD analysis using filtration or centrifugation.

In some embodiments slow cooling was used. In this method a concentrated suspension of compound in solvent is provided. This 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 collected into clean vials. The resulting clear solution was cooled to 5° C. to induce precipitation. Solids were isolated solids for XRPD analysis using filtration or centrifugation.

In some embodiments crash cooling was used. In this method a concentrated suspension of compound in solvent is provided. 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 collected into clean vials. The clear solution was cooled to −20° C. to induce precipitation. Solids were isolated for XRPD analysis using filtration or centrifugation.

In some embodiments liquid vapor diffusion was used. In this method a concentrated stock of compound in solvent is provided. This concentrated stock is transferred to an inner vial that is sealed within a larger vial containing anti-solvent. Solids were isolated for XRPD analysis using filtration or centrifugation.

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

Unique HM30181 mesylate polymorphisms were characterized by a variety of techniques, including X-ray powder diffraction (XRPD), NMR, and calorimetry. These were performed as follows.

XRPD was performed using a Panalytical X'Pert3™ Powder XRPD and on a Si zero-background holder. The 2θ position was calibrated against a Panalytical™ 640 Si powder standard. Details of XRPD used in the experiments are listed below in Table 20.

TABLE 20Parameters for Reflection ModeX-Ray wavelengthCu, kα, Kα1 (Å): 1.540598,Kα2 (Å): 1.544426Kα2/Kα1 intensity ratio: 0.50X-Ray tube setting45 kV, 40 mADivergence slitAutomaticScan modeContinuousScan range (°2TH)3°-40°Step size (°2TH)0.0131Scan speed (°/s)0.16

Differential Scanning calorimetry (DSC) was performed using a TA Q2000™ DSC from TA Instruments. Temperature was ramped from ambient temperature to desired temperature at a heating rate of 10° C./min using N2as the purge gas, with pan crimped (see Table 21).

TABLE 21ParametersDSCPan TypeAluminum pan, closedTemperatureambient temperature-300° C.Ramp rate10° C./minPurge gasN2
In some studies, a cyclic DSC method was used. In such cycling DSC methods temperature was ramped from ambient to 150° C. at a heating rate of 10° C./min using N2as the purge gas, then cooled by 10° C. to 25° C. This temperature cycle repeated twice (see Table 22).

TABLE 22ParametersDSCPan TypeAluminum pan, closedTemperature25-150°C.Ramp rate10°C./minPurge gasN2Heat/cool Cycles2

Thermogravimetric Analysis (TGA) was performed using a TA Q500™ TGA from TA Instruments. Temperature was ramped from ambient to desired temperature at a heating rate of 10° C./min using N2as the purge gas, with pan open (see Table 23).

TABLE 23ParametersTGAPan TypePlatinum plate, openTemperature300°C.Ramp rate10°C./minPurge gasN2Sample purge flow15mL/minBalance purge flow25mL/min

Dynamic Vapor Sorption (DVS) was measured using a SMS (Surface Measurement Systems™) DVS Intrinsic. Parameters for DVS test are listed below in Table 24.

TABLE 24ParametersValuesTemperature25°C.Sample size10-20mgGas and flow rateN2, 200 mL/mindm/dt0.002%/minMin. dm/dt stability duration10minMax. equilibrium time360minRH range40% RH-95% RH-0% RH-95% RHRH step size10%(0% RH-90% RH)5%(90% RH-95% RH)

Proton NMR were obtained using a Varian 200M™ NMR in deuterated DMSO (DMSO-d6).

It should be apparent to those skilled in the art that many more modifications besides those already described are possible without departing from the inventive concepts herein. The inventive subject matter, therefore, is not to be restricted except in the spirit of the appended claims. Moreover, in interpreting both the specification and the claims, all terms should be interpreted in the broadest possible manner consistent with the context. In particular, the terms “comprises” and “comprising” should be interpreted as referring to elements, components, or steps in a non-exclusive manner, indicating that the referenced elements, components, or steps may be present, or utilized, or combined with other elements, components, or steps that are not expressly referenced. Where the specification claims refer to at least one of something selected from the group consisting of A, B, C . . . and N, the text should be interpreted as requiring only one element from the group, not A plus N, or B plus N, etc.