Polymorphisms of HM30181 mesylate

Abstract

HM30181 (shown below) can be used to improve absorption of cancer chemotherapy drugs, such as paclitaxel. Herein are described various polymorphisms of HM30181, in particular polymorphisms B, D, E, F, G, H, I, J, K, L, and M as well as their physical properties and methods for their preparation and characterization.

Description

FIELD OF THE INVENTION

The field of the invention is P-glycoprotein inhibitors, in particular HM30181 mesylate.

BACKGROUND

The background 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.

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

Unfortunately, a number of drugs utilized in chemotherapy are substrates for P-gp. P-gp activity, therefore, can reduce bioavailability and effectiveness of chemotherapeutic drugs. In such instances administration of P-gp inhibitors can be useful in improving the response to chemotherapy. Accordingly, over the last 30 years a number of pharmaceutically useful P-gp inhibitors (such as amiodarone, clarithromycin, cyclosporin, colchicine, diltiazem, erythromycin, felodipine, ketoconazole, lansoprazole, omeprazole, nifedipine, paroxetine, reserpine, saquinavir, sertraline, quinidine, tamoxifen, verapamil, and duloxetine) have been developed.

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

The pharmacokinetics, bioavailability, and incidence of side effects of orally administered HM30181 mesylate are less than optimal, however.

Thus, there is still a need for polymorphisms of HM30181 mesylate that can provide improved absorption, improved pharmacokinetics, and/or reduced side effects upon administration.

SUMMARY OF THE INVENTION

The inventive subject matter provides polymorphisms of HM31081 mesylate, methods for their preparation and characterization, and methods for their use.

One embodiment of the inventive concept is a composition that includes a crystalline or partially crystalline form of HM30181 mesylate, where 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. In some of such embodiments the crystalline or partially crystalline form is polymorph B, and has an X-ray diffraction pattern corresponding to FIG. 40 and an endotherm at about 159.92° C. In some of such embodiments the crystalline or partially crystalline form is polymorph Type C, and has an X-ray diffraction pattern corresponding to FIG. 42 and an endotherm at about 159.6° C. In some of such embodiments the crystalline or partially crystalline form is polymorph Type C, and the crystalline form has an X-ray diffraction pattern with 2Theta maxima at about 6.4° and about 8.0°. In some of such embodiments the crystalline or partially crystalline form is polymorph Type C, and has a unit cell with a volume of about 1180 ų, where a is about 7 Å, b is about 15 Å, c is about 18 Å, alpha is about 52°, beta is about 62°, and gamma is about 90°. In some of such embodiments the crystalline or partially crystalline form is polymorph C, and has an X-ray diffraction pattern corresponding to FIG. 42 and an endotherm at about 159.60° C., and can be a monohydrate. In some of such embodiments the crystalline or partially crystalline form is polymorph D, and has an X-ray diffraction pattern corresponding to FIG. 45 and an endotherm at about 66.97° C. In some of such embodiments the crystalline or partially crystalline form is polymorph E, and has an X-ray diffraction pattern corresponding to FIG. 47 and an endotherm at about 154.42° C., and can include DMA. In some of such embodiments the crystalline or partially crystalline form is polymorph Type E, and has an X-ray diffraction pattern corresponding to FIG. 47 and an endotherm at about 154.4° C. In some of such embodiments the crystalline or partially crystalline form is polymorph Type E, and has an X-ray diffraction pattern comprising 2Theta 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 of such embodiments the crystalline or partially crystalline form is polymorph Type E, and has a unit cell with a volume of about 1620 ų, where a is about 8 Å, b is about 10 Å, c is about 24 Å, alpha is about 75°, beta is about 80°, and gamma is about 110°. In some of such embodiments the crystalline or partially crystalline form is polymorph F, and has an X-ray diffraction pattern corresponding to FIG. 50 has an endotherm at about 148.41° C., and can include DMF. In some of such embodiments the crystalline or partially crystalline form is polymorph G, and has an X-ray diffraction pattern corresponding to FIG. 53 and an endotherm at about 69.02° C. In some of such embodiments the crystalline or partially crystalline form is polymorph H, and has an X-ray diffraction pattern corresponding to FIG. 55 and an endotherm at about 126.52° C. In some of such embodiments the crystalline or partially crystalline form is polymorph I, and has an X-ray diffraction pattern corresponding to FIG. 57. In some of such embodiments the crystalline or partially crystalline form is polymorph J, and has an X-ray diffraction pattern corresponding to FIG. 58. In some of such embodiments the crystalline or partially crystalline form is polymorph K, and has an X-ray diffraction pattern corresponding to FIG. 60. In some of such embodiments the crystalline or partially crystalline form is polymorph L, and has an X-ray diffraction pattern corresponding to FIG. 61. In some of such embodiments the crystalline or partially crystalline form is polymorph M, and has an X-ray diffraction pattern corresponding to FIG. 62. In some of such embodiments the crystalline or partially crystalline form is polymorph N, has an X-ray diffraction pattern corresponding to FIG. 63 and an endotherms at about 159° C. and about 188° C., and can include methanol.

Another embodiment of the inventive concept is a method of inhibiting P-glycoprotein activity, by contacting P-glycoprotein with at least one crystalline or partially crystalline form of HM30181 mesylate as described above in an amount that is effective in inhibiting an activity of P-glycoprotein.

Another embodiment of the inventive concept is a method of treating cancer by administering a chemotherapeutic drug that is a P-glycoprotein substrate to an individual that in need of treatment for cancer and also administering a polymorph of HM30181 mesylate as described above in an amount that is effective to inhibit P-glycoprotein activity in the individual.

Another embodiment of the inventive concept is the use of a polymorph of HM30181 mesylate as described above in preparing a medicament for treating cancer. Such a medicament can also include a chemotherapeutic drug that is a P-glycoprotein substrate.

Another embodiment of the inventive concept is a formulation that includes a polymorph of HM30181 mesylate as described above and a therapeutic drug, where the therapeutic drug is a P-glycoprotein substrate. Such a therapeutic drug can be a chemotherapeutic drug that is used in the treatment of cancer.

Various objects, features, aspects and advantages of the inventive subject matter will become more apparent from the following detailed description of preferred embodiments, along with the accompanying drawing figures in which like numerals represent like components.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1: FIG. 1 shows an overlay of X-Ray Powder Diffraction (XRPD) results from HM30181 mesylate polymorphisms types A to N.

FIG. 2: FIG. 2 shows results from XRPD studies of HM30181 mesylate Type A starting material

FIG. 3: FIG. 3 shows an overlay of results of DSC/TGA studies of HM30181 mesylate Type A.

FIG. 4: FIG. 4 shows results of 1H-NMR studies of HM30181 mesylate Type A.

FIG. 5: FIG. 5 shows results of DVS of HM30181 mesylate Type A starting material.

FIG. 6: FIG. 6 shows an overlay of overlay of XRPD studies of Type A starting material post-DVS.

FIG. 7: FIG. 7 shows an overlay of results from Cyclic DSC studies of HM30181 mesylate Type A and HM30181 mesylate Type A heated to 110° C.

FIG. 8: FIG. 8 shows an overlay of results of XRPD studies of HM30181 mesylate Type A and HM30181 mesylate Type A heated to 110° C.

FIG. 9: FIG. 9 shows an overlay of cyclic DSC studies of HM30181 mesylate Type A starting material and HM30181 mesylate Type A heated to 185° C.

FIG. 10: FIG. 10 shows an overlay of XRPD studies of HM30181 mesylate Type A starting material and HM30181 mesylate Type A heated to 185° C.

FIG. 11: FIG. 11 shows an overlay of results of cyclic DSC studies of HM30181 mesylate Type A starting material and HM30181 mesylate Type A heated to 200° C.

FIG. 12: FIG. 12 shows an overlay of results of XRPD studies of HM30181 mesylate Type A starting material and HM30181 mesylate Type A heated to 200° C.

FIG. 13: FIG. 13 shows results of XRPD studies of freebase HM3018-A

FIG. 14: FIG. 14 shows an overlay of results of ¹H-NMR studies of HM30181 mesylate Type A heated to 200° C., freebase HM3018-A, and HM30181 mesylate Type A starting material.

FIG. 15: FIG. 15 shows an overlay of results of XRPD studies of HM30181 mesylate Type B from slurry experiments.

FIG. 16: FIG. 16 shows an overlay of results of XRPD studies of HM30181 mesylate Type C from slurry experiments.

FIG. 17: FIG. 17 shows an overlay of results of XRPD studies of HM30181 mesylate Type D from slurry experiments.

FIG. 18: FIG. 18 shows an overlay of results of XRPD studies of HM30181 mesylate Type E from slurry experiments.

FIG. 19: FIG. 19 shows an overlay of results of XRPD of HM30181 mesylate Type F from slurry experiments.

FIG. 20: FIG. 20 shows an overlay of results of XRPD studies of HM30181 mesylate Type G from slurry experiments.

FIG. 21: FIG. 21 shows an overlay of results of XRPD studies of HM30181 mesylate Type D from liquid vapor diffusion experiments.

FIG. 22: FIG. 22 shows an overlay of results of XRPD studies of HM30181 mesylate Type E from liquid vapor diffusion experiments.

FIG. 23: FIG. 23 shows an overlay of results of XRPD studies of HM30181 mesylate Type H from liquid vapor diffusion experiments.

FIG. 24: FIG. 24 shows an overlay of results of XRPD studies of HM30181 mesylate Type I and HM30181 mesylate Type J from liquid vapor diffusion experiments.

FIG. 25: FIG. 25 shows an overlay of results of XRPD studies of HM30181 mesylate Type C polymorph generated by cooling.

FIG. 26: FIG. 26 shows an overlay of results of XRPD studies of HM30181 mesylate Type C with samples from anti-solvent experiments.

FIG. 27: FIG. 27 shows an overlay of results of XRPD studies of HM30181 mesylate Type F with sample from anti-solvent experiments.

FIG. 28: FIG. 28 shows an overlay of results of XRPD studies of HM30181 mesylate Type J with samples from anti-solvent experiments.

FIG. 29: FIG. 29 shows an overlay of results of XRPD studies of HM30181 mesylate Type K from anti-solvent experiments.

FIG. 30: FIG. 30 shows an overlay of results of XRPD studies of HM30181 mesylate Type L from anti-solvent experiments.

FIG. 31: FIG. 31 shows an overlay of results of XRPD studies of HM30181 mesylate Type M from anti-solvent experiments.

FIG. 32: FIG. 32 shows an overlay of results of XRPD studies of HM30181 mesylate Type B large scale studies.

FIG. 33: FIG. 33 shows an overlay of results of XRPD studies of HM30181 mesylate Type C large scale studies.

FIG. 34: FIG. 34 shows an overlay of XRPD studies of HM30181 mesylate Type D large scale studies.

FIG. 35: FIG. 35 shows an overlay of results of XRPD studies of HM30181 mesylate Type E large scale studies.

FIG. 36: FIG. 36 shows an overlay of results of XRPD studies of HM30181 mesylate Type G large scale studies.

FIG. 37: FIG. 37 shows an overlay of results of XRPD studies of HM30181 mesylate Type F and HM30181 mesylate Type G large scale studies.

FIG. 38: FIG. 38 shows an overlay of results of 1H-NMR studies of HM30181 mesylate Type G large scale studies.

FIG. 39: FIG. 39 shows an overlay of results of 1H-NMR studies HM30181 mesylate polymorphisms large scale studies.

FIG. 40: FIG. 40 shows results of XRPD studies of HM30181 mesylate Type B.

FIG. 41: FIG. 41 shows an overlay of DSC and TGA results from HM30181 mesylate Type B.

FIG. 42: FIG. 42 shows results of XRPD of HM30181 mesylate Type C.

FIG. 43: FIG. 43 shows an overlay of DSC and TGA results of HM30181 mesylate Type C.

FIG. 44: FIG. 44 shows results of 1H-NMR of HM30181 mesylate Type C.

FIG. 45: FIG. 45 shows results of XRPD of HM30181 mesylate Type D.

FIG. 46: FIG. 46 shows an overlay of DSC and TGA results from HM30181 mesylate Type D.

FIG. 47: FIG. 47 shows results of XRPD of HM30181 mesylate Type E.

FIG. 48: FIG. 48 shows an overlay of results from DSC and TGA of HM30181 mesylate Type E.

FIG. 49: FIG. 49 shows results of 1H-NMR of HM30181 mesylate Type E.

FIG. 50: FIG. 50 shows results from XRPD of HM30181 mesylate Type F.

FIG. 51: FIG. 51 shows an overlay of results from DSC and TGA of HM30181 mesylate Type F.

FIG. 52: FIG. 52 shows results from 1H-NMR of HM30181 mesylate Type F

FIG. 53: FIG. 53 shows results from XRPD of HM30181 mesylate Type G.

FIG. 54: FIG. 54 shows an overlay of results from DSC and TGA of HM30181 mesylate Type G.

FIG. 55: FIG. 55 shows results from XRPD of HM30181 mesylate Type H.

FIG. 56: FIG. 56 shows an overlay of results from DSC and TGA of HM30181 mesylate Type H.

FIG. 57: FIG. 57 shows results from XRPD of HM30181 mesylate Type I.

FIG. 58: FIG. 58 shows results from XRPD of HM30181 mesylate Type J.

FIG. 59: FIG. 59 shows results from TGA of HM30181 mesylate Type J.

FIG. 60: FIG. 60 shows results from XRPD of HM30181 mesylate Type K.

FIG. 61: FIG. 61 shows results from XRPD of HM30181 mesylate Type L.

FIG. 62: FIG. 62 shows results from XRPD of HM30181 mesylate Type M.

FIG. 63: FIG. 63 shows results from XRPD of HM30181 mesylate Type N.

FIG. 64: FIG. 64 shows an overlay of DSC and TGA results from HM30181 mesylate Type N.

FIG. 65: FIG. 65 shows results of 1H-NMR of HM30181 mesylate Type N.

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 0
	Crystal	DSC
	form	endotherm
Batch	(XRPD)	(° C.)	TGA weight loss	Identification
B00505-05-C	Type A	181.41	2.6% before 150° C.	Monohydrate (hygroscopic
(starting material)				to 2.26% at 95% RH)
6007235-16-A1-airdry	Type B	159.92	1.9% before 170° C.	Solvate/hydrate
6004273-10-C-vd	Type C	159.60	3.0% before 175° C.	Monohydrate
6007235-16-A21-airdry	Type D	66.97 (peak),	14.71% before 150° C.	Solvate/hydrate
		114 (exotherm)
6004273-10-E-vd	Type E	154.42	5.247% before 168° C.	DMA solvate
6004273-10-G-vd	Type F	148.41	5.123% weight loss	DMF solvate
			before 180° C.
6007235-16-B26-airdry	Type G	69.02	1.451% before 150° C.	Solvate/hydrate
		233.29
6007235-19-B10	Type H	126.52	7.444% weight loss	Solvate/hydrate
			before 200° C.
6007235-19-E3	Type I	NA	NA	Weak solvate- converts to
				Type J on air-drying
6007235-19-E5	Type J	NA	2.887% weight loss	Solvate/hydrate
			before 150° C.
6004273-06-A4	Type K	Insufficient	Insufficient sample	Possible solvate/hydrate
		sample
6004273-06-A60	Type L	Insufficient	Insufficient sample	Possible solvate/hydrate
		sample
6004273-06-A56	Type M	Insufficient	Insufficient sample	Possible solvate/hydrate
		sample
6004273-10-B-vd	Type N	159.28	2.128% weight loss	Possible MeOH
		188.47	before 180° C.	solvate/hydrate

FIG. 1 provides 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 (see FIG. 2).

By DSC, HM30181 mesylate Type A displayed an endotherm at 181.41° C. (see FIG. 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 (see FIG. 3). By ¹H-NMR, HM30181 mesylate Type A was potentially a hydrate, as no solvents were detected other than water (see FIG. 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 (see FIG. 5 and FIG. 6).

Cyclic DSC to 110° C. of HM30181 mesylate Type A resulted in no XRPD change (see FIG. 7 and FIG. 8). Cyclic DSC of HM30181 mesylate Type A to 185° C. resulted in HM30181 mesylate Type A with reduced crystallinity (FIG. 9 and FIG. 10). Cyclic DSC of HM30181 mesylate Type A to 200° C. resulted in amorphous material (see FIG. 11 and FIG. 12). Freebase HM3018-A (6004273-06-A) was also characterized by XRPD and was found to be amorphous (see FIG. 13). Based on ¹H-NMR data, decomposition of Type A initiated after heating to 200° C. (see FIG. 14).

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

TABLE 2
			Dissolved after
		Solubility	heating to 50° C.
Experiment ID	Solvent	(mg/mL)	for 2 hours?
B00505-05-C-1	MeOH	1.7-2.0
B00505-05-C-2	EtOH	<0.9	Almost
B00505-05-C-3	IPA	<1.0	No
B00505-05-C-4	Acetone	<1.0	No
B00505-05-C-5	MIBK	<1.0	No
B00505-05-C-6	EtOAc	<0.9	No
B00505-05-C-7	IPOAc	<1.0	No
B00505-05-C-8	THF	<1.0	No
B00505-05-C-9	2-MeTHF	<1.0	No
B00505-05-C-10	1,4-Dioxane	<1.0	No
B00505-05-C-11	MTBE	<1.0	No
B00505-05-C-12	ACN	<1.0	Yes
B00505-05-C-13	DCM	<1.0	Yes
B00505-05-C-14	CHCl3	5.0-6.7
B00505-05-C-15	Toluene	<1.0	No
B00505-05-C-16	n-Heptane	<1.0	No
B00505-05-C-17	H2O	<1.0	No
B00505-05-C-18	Cyclohexane	<1.0	No
B00505-05-C-19	MEK	<1.0	No
B00505-05-C-20	T-BuOH	<1.0	No
B00505-05-C-21	NMP	4.8-6.3
B00505-05-C-22	DMSO	22-44
B00505-05-C-23	DMA	3.2-4.8
B00505-05-C-24	n-Propanol	<1.0	No
B00505-05-C-25	n-Propyl acetate	<1.0	No
B00505-05-C-26	Cumene	<1.0	No

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. (see FIG. 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 (see FIG. 16). Slurries of HM30181 mesylate Type A in NMP at ambient temperature generated HM30181 mesylate Type D (see FIG. 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 (see FIG. 18). Slurries of HM30181 mesylate Type A in DMF generated HM30181 mesylate Type F at ambient temperature (see FIG. 19) and Type G at 50° C. (see FIG. 20).

Table 3 (continued in Table 4)
	API	Solvent	Crystal
Exp	Method	Temp.	Solvent	(mg)	(mL)	Type
6007235-16-C10	Slurry	4°	C.	1,4-Dioxane	25.9	0.2	A
6007235-16-B10	Slurry	50°	C.	1,4-Dioxane	23.4	0.2	A
6007235-16-A10	Slurry	ambient	1,4-Dioxane	21	0.2	A
		temperature
6007235-16-A34	Slurry	ambient	1-Methyl-2-	25	0.2	A
		temperature	pyrrolidinone/water
			(1:1)
6007235-16-C9	Slurry	4°	C.	2-Methyl	27.4	0.2	A
				tetrahydrofuran
6007235-16-B9	Slurry	50°	C.	2-Methyl	24.5	0.2	A
				tetrahydrofuran
6007235-16-A9	Slurry	ambient	2-Methyl	28.7	0.2	A
		temperature	tetrahydrofuran
6007235-16-C4	Slurry	4°	C.	Acetone	25.3	0.2	A
6007235-16-B4	Slurry	50°	C.	Acetone	27.9	0.2	A
6007235-16-A4	Slurry	ambient	Acetone	26.7	0.2	A
		temperature
6007235-16-C13	Slurry	4°	C.	CHCl3	22.4	0.2	A
6007235-16-B13	Slurry	50°	C.	CHCl3	29	0.1	A
6007235-16-A14	Slurry	ambient	CHCl3	25.6	0.2	A
		temperature
6007235-16-C21	Slurry	4°	C.	Cumene	22.1	0.2	A
6007235-16-B25	Slurry	50°	C.	Cumene	26.3	0.2	A
6007235-16-A26	Slurry	ambient	Cumene	26.1	0.2	A
		temperature
6007235-16-C17	Slurry	4°	C.	Cyclohexane	24.2	0.2	A
6007235-16-B17	Slurry	50°	C.	Cyclohexane	23.8	0.2	A
6007235-16-A18	Slurry	ambient	Cyclohexane	26.2	0.2	A
		temperature
6007235-16-C24	Slurry	4°	C.	Cyclopentylmethyl	20.8	0.2	A
				ether
6007235-16-B29	Slurry	50°	C.	Cyclopentylmethyl	24.9	0.2	A
				ether
6007235-16-A30	Slurry	ambient	Cyclopentylmethyl	21.7	0.2	A
		temperature	ether
6007235-16-A22	Slurry	ambient	DMSO	22.7	0.1	A
		temperature
6007235-16-A31	Slurry	ambient	DMSO/water (1:1)	25.1	0.2	A
		temperature
6007235-16-C2	Slurry	4°	C.	Ethanol	20.7	0.2	A
6007235-16-B2	Slurry	50°	C.	Ethanol	26.9	0.2	A
6007235-16-A2	Slurry	ambient	Ethanol	21.7	0.2	A
		temperature
6007235-16-C6	Slurry	4°	C.	Ethyl acetate	26.2	0.2	A
6007235-16-B6	Slurry	50°	C.	Ethyl acetate	25.5	0.2	A
6007235-16-A6	Slurry	ambient	Ethyl acetate	24.2	0.2	A
		temperature
6007235-16-C22	Slurry	4°	C.	Ethyl formate	24.8	0.2	A
6007235-16-B27	Slurry	50°	C.	Ethyl formate	25	0.2	A
6007235-16-A28	Slurry	ambient	Ethyl formate	25.1	0.2	A
		temperature
6007235-16-C23	Slurry	4°	C.	Isobutyl Acetate	23.7	0.2	A
6007235-16-B28	Slurry	50°	C.	Isobutyl Acetate	22	0.2	A
6007235-16-A29	Slurry	ambient	Isobutyl Acetate	25.4	0.2	A
		temperature
6007235-16-C3	Slurry	4°	C.	isopropanol	26.8	0.2	A
6007235-16-B3	Slurry	50°	C.	isopropanol	21.5	0.2	A
6007235-16-A3	Slurry	ambient	isopropanol	23.6	0.2	A
		temperature
6007235-16-C7	Slurry	4°	C.	isopropyl acetate	25.9	0.2	A
6007235-16-B7	Slurry	50°	C.	isopropyl acetate	21.9	0.2	A
6007235-16-A7	Slurry	ambient	isopropyl acetate	28.8	0.2	A
		temperature
6007235-16-C18	Slurry	4°	C.	methyl ethyl	21.8	0.2	A
				ketone
6007235-16-B18	Slurry	50°	C.	methyl ethyl	20.6	0.2	A
				ketone
6007235-16-A19	Slurry	ambient	methyl ethyl	27.1	0.2	A
		temperature	ketone
6007235-16-C5	Slurry	4°	C.	methyl isobutyl	26.1	0.2	A
				ketone
6007235-16-B5	Slurry	50°	C.	methyl isobutyl	27	0.2	A
				ketone
6007235-16-A5	Slurry	ambient	methyl isobutyl	27.9	0.2	A
		temperature	ketone
Table 0 (continued from Table 3)
	API	Solvent	Crystal
Exp	Method	Temp.	Solvent	(mg)	(mL)	Type
6007235-16-C11	Slurry	4°	C.	Methyl t-butyl ether	27.1	0.2	A
6007235-16-B11	Slurry	50°	C.	Methyl t-butyl ether	27.5	0.2	A
6007235-16-A11	Slurry	ambient	Methyl t-butyl ether	26.8	0.2	A
		temperature
6007235-16-A32	Slurry	ambient	N,N-	26.9	0.2	A
		temperature	Dimethylacetamide/water
			(1:1)
6007235-16-A33	Slurry	ambient	N,N-	27	0.2	A
		temperature	Dimethylacetamide/water
			(1:1)
6007235-16-C15	Slurry	4°	C.	n-Heptane	26.8	0.2	A
6007235-16-B15	Slurry	50°	C.	n-Heptane	22.7	0.2	A
6007235-16-A16	Slurry	ambient	n-Heptane	22.8	0.2	A
		temperature
6007235-16-C19	Slurry	4°	C.	n-Propanol	22.3	0.2	A
6007235-16-B23	Slurry	50°	C.	n-Propanol	21.8	0.2	A
6007235-16-A24	Slurry	ambient	n-Propanol	23.3	0.2	A
		temperature
6007235-16-C20	Slurry	4°	C.	n-Propyl acetate	21.5	0.2	A
6007235-16-B24	Slurry	50°	C.	n-Propyl acetate	27.4	0.2	A
6007235-16-A25	Slurry	ambient	n-Propyl acetate	26.5	0.2	A
		temperature
6007235-16-B19	Slurry	50°	C.	t-Butanol	23.7	0.2	A
6007235-16-A20	Slurry	ambient	t-Butanol	23.9	0.2	A
		temperature
6007235-16-C8	Slurry	4°	C.	Tetrahydrofuran	22.9	0.2	A
6007235-16-B8	Slurry	50°	C.	Tetrahydrofuran	21.7	0.2	A
6007235-16-A8	Slurry	ambient	Tetrahydrofuran	20.9	0.2	A
		temperature
6007235-16-C14	Slurry	4°	C.	Toluene	24.2	0.2	A
6007235-16-B14	Slurry	50°	C.	Toluene	25.7	0.2	A
6007235-16-A15	Slurry	ambient	Toluene	24.9	0.2	A
		temperature
6007235-16-C16	Slurry	4°	C.	Water	27.6	0.2	A
6007235-16-B16	Slurry	50°	C.	Water	27	0.2	A
6007235-16-C28	Slurry	4°	C.	1-Methyl-2-	23.9	0.2	amorphous +
				pyrrolidinonee/water			A
				(1:1)
6007235-16-C26	Slurry	4°	C.	N,N-	26.4	0.2	amorphous +
				Dimethylacetamide/water			A
				(1:1)
6007235-16-C27	Slurry	4°	C.	N,N-	24	0.2	amorphous +
				Dimethylacetamide/water			A
				(1:1)
6007235-16-A17	Slurry	ambient	Water	22	0.2	amorphous +
		temperature				A
6007235-16-B20	Slurry	50°	C.	1-Methyl-2-pyrrolidinone	23.7	0.1	amorphous
6007235-16-B21	Slurry	50°	C.	DMSO	23.8	0.1	amorphous
6007235-16-C25	Slurry	4°	C.	DMSO/water (1:1)	24.3	0.2	amorphous
6007235-16-A1	Slurry	ambient	Methanol	24.9	0.2	B
		temperature
6007235-16-B1	Slurry	50°	C.	Methanol	27	0.1	B
6007235-16-C1	Slurry	4°	C.	Methanol	23.2	0.2	B
6007235-16-A12	Slurry	ambient	Acetonitrile	25.5	0.2	C
		temperature
6007235-16-A13	Slurry	ambient	Dichloromethane	21.9	0.2	C
		temperature
6007235-16-B12	Slurry	50°	C.	Acetonitrile	22.7	0.1	C
6007235-16-C12	Slurry	4°	C.	Acetonitrile	21.9	0.2	C
6007235-16-A21	Slurry	ambient	1-Methyl-2-pyrrolidinone	24.6	0.2	D
		temperature
6007235-16-A23	Slurry	ambient	N,N-Dimethylacetamide	25.6	0.1	E
		temperature
6007235-16-A27	Slurry	ambient	N,N-Dimethylacetamide	26.2	0.1	F
		temperature
6007235-16-B26	Slurry	50°	C.	N,N-Dimethylacetamide	27.3	0.1	G

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 (see FIG. 21). Liquid vapor diffusion of MEK into NMP or DMA solution yielded HM30181 mesylate Type D (see FIG. 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 (see FIG. 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 (see FIGS. 4 to 8). Liquid vapor diffusion of ACN into DMSO solution yielded HM30181 mesylate Type H (see FIG. 23). Liquid vapor diffusion of Acetone, Ethyl Acetate, or isopropyl acetate into DMA solution yielded HM30181 mesylate Type I (see FIG. 24). Air-drying of HM30181 mesylate Type I yielded HM30181 mesylate Type J (see FIG. 24). Liquid vapor diffusion of isopropyl acetate into DMSO solution followed by air drying also yielded HM30181 mesylate Type J.

TABLE 5
			API	Solvent	Crystal
Exp	Temperature	Solvent	(mg)	(mL)	Type
6007235-17-A1	ambient	methanol	25.2	0.1	A
	temperature
6007235-17-A2	ambient	ethanol	22.1	0.1	A
	temperature
6007235-17-A3	ambient	isopropanol	21.4	0.1	A
	temperature
6007235-17-A4	ambient	acetone	24.8	0.1	A
	temperature
6007235-17-A5	ambient	methyl isobutyl ketone	25.8	0.1	A
	temperature
6007235-17-A6	ambient	ethyl acetate	22.9	0.1	A
	temperature
6007235-17-A7	ambient	isopropyl acetate	23.8	0.1	A
	temperature
6007235-17-A8	ambient	tetrahydrofuran	24.5	0.1	A
	temperature
6007235-17-A9	ambient	2-methyl tetrahydrofuran	24.9	0.1	A
	temperature
6007235-17-A10	ambient	methyl t-butyl ether	27.1	0.1	A
	temperature
6007235-17-A11	ambient	acetonitrile	27.7	0.1	A
	temperature
6007235-17-A12	ambient	dichloromethane	24.9	0.1	A
	temperature
6007235-17-A13	ambient	CHCl3	29	0.1	A
	temperature
6007235-17-A14	ambient	methyl ethyl ketone	23.4	0.1	A
	temperature
6007235-17-A15	ambient	t-butanol	28.2	0.1	A
	temperature
6007235-17-A16	ambient	n-propanol	25.4	0.1	A
	temperature
6007235-17-A17	ambient	ethyl formate	27.3	0.1	A
	temperature
6007235-17-A18	ambient	cyclopentylmethyl ether	25.1	0.1	A
	temperature
6007235-17-A19	25° C.	25° C./60% RH	22.9	0.1	A
6007235-17-A20	40° C.	40° C./75% RH	23.5	0.1	A
6007235-17-A21	40° C.	100% RH(water)	22.7	0.1	A

Table 6 (continued on Table 7)
						Anti-
			API	Solvent		solvent
Exp	Temp.	Solvent	(mg)	(mL)	Anti-solvent	(mL)	Crystal Type
6007235-19-A1	ambient	methanol	22	16	ethyl formate	0.5	No solids
	temperature
6007235-19-A2	ambient	methanol	23	16	dichloromethane	0.5	No solids
	temperature
6007235-19-B1	ambient	DMSO	25	0.3	ethanol	2	No solids
	temperature
6007235-19-B2	ambient	DMSO	25	0.3	isopropanol	2	No solids
	temperature
6007235-19-B3	ambient	DMSO	25	0.3	acetone	2	Amorphous
	temperature
6007235-19-B4	ambient	DMSO	25	0.3	MIBK	2	Amorphous
	temperature
6007235-19-B5	ambient	DMSO	25	0.3	ethyl acetate	2	Amorphous
	temperature
6007235-19-B7	ambient	DMSO	25	0.3	tetrahydrofuran	2	Amorphous
	temperature
6007235-19-B8	ambient	DMSO	25	0.3	2-MeTHF	2	Amorphous
	temperature
6007235-19-B9	ambient	DMSO	25	0.3	methyl t-butyl	2	Amorphous
	temperature				ether
6007235-19-B11	ambient	DMSO	25	0.3	dichloromethane	2	Amorphous
	temperature						and A
6007235-19-B12	ambient	DMSO	25	0.3	methyl ethyl	2	Amorphous
	temperature				ketone
6007235-19-B13	ambient	DMSO	25	0.3	t-butanol	2	No solids
	temperature
6007235-19-C1	ambient	NMP	30	3	ethanol	3	No solids
	temperature
6007235-19-C2	ambient	NMP	30	3	isopropanol	3	No solids
	temperature
6007235-19-C3	ambient	NMP	30	3	acetone	3	No solids
	temperature
6007235-19-C4	ambient	NMP	30	3	MIBK	3	No solids
	temperature
6007235-19-C5	ambient	NMP	30	3	ethyl acetate	3	Amorphous
	temperature						and A
6007235-19-C6	ambient	NMP	30	3	isopropyl acetate	3	No solids
	temperature
6007235-19-C7	ambient	NMP	30	3	tetrahydrofuran	3	No solids
	temperature
6007235-19-C8	ambient	NMP	30	3	2-MeTHF	3	No solids
	temperature
6007235-19-C10	ambient	NMP	30	3	acetonitrile	3	No solids
	temperature
6007235-19-C11	ambient	NMP	30	3	dichloromethane	3	No solids
	temperature
6007235-19-C13	ambient	NMP	30	3	t-butanol	3	No solids
	temperature
6007235-19-D1	ambient	DMF	30	3	ethanol	3	No solids
	temperature
6007235-19-D2	ambient	DMF	30	3	isopropanol	3	Amorphous
	temperature
6007235-19-D4	ambient	DMF	30	3	MIBK	3	No solids
	temperature
6007235-19-D5	ambient	DMF	30	3	ethyl acetate	3	Amorphous
	temperature
6007235-19-D6	ambient	DMF	30	3	isopropyl acetate	3	Amorphous
	temperature
6007235-19-D7	ambient	DMF	30	3	tetrahydrofuran	3	No solids
	temperature
6007235-19-D9	ambient	DMF	30	3	methyl t-butyl	3	Amorphous
	temperature				ether
6007235-19-D10	ambient	DMF	30	3	acetonitrile	3	No solids
	temperature
6007235-19-D11	ambient	DMF	30	3	dichloromethane	3	No solids
	temperature
6007235-19-D12	ambient	DMF	30	3	methyl ethyl	3	Amorphous
	temperature				ketone
6007235-19-D13	ambient	DMF	30	3	t-butanol	3	No solids
	temperature
6007235-19-E1	ambient	DMA	20	4	ethanol	8	No solids
	temperature
6007235-19-E2	ambient	DMA	20	4	isopropanol	8	No solids
	temperature
6007235-19-E4	ambient	DMA	20	4	MIBK	8	No solids
	temperature
6007235-19-E7	ambient	DMA	20	4	tetrahydrofuran	8	No solids
	temperature
6007235-19-E10	ambient	DMA	20	4	acetonitrile	8	No solids
	temperature
6007235-19-E11	ambient	DMA	20	4	dichloromethane	8	No solids
	temperature
6007235-19-E13	ambient	DMA	20	4	t-butanol	8	No solids
	temperature
Table 7 (continued from Table 6)
						Anti-	XRPD
		API		Solvent		solvent	(wet	XRPD
Exp.	Temp.	(mg)	Solvent	(mL)	Anti-solvent	(mL)	cake)	(air-dry)
6007235-19-C12	ambient	30	NMP	3	MEK	3	Type D	—
	temperature
6007235-19-E12	ambient	20	DMA	4	MEK	8	Type D	Type D and
	temperature							amorphous
6007235-19-C9	ambient	30	NMP	3	MTBE	3	Type D	—
	temperature
6007235-19-D8	ambient	30	DMF	3	2-MeTHF	3	Type E	E
	temperature
6007235-19-E8	ambient	20	DMA	4	2-MeTHF	8	Type E	E
	temperature
6007235-19-E9	ambient	20	DMA	4	MTBE	8	Type E	E
	temperature						and I
6007235-19-B10	ambient	25	DMSO	0.3	ACN	2	Type H	—
	temperature
6007235-19-E3	ambient	20	DMA	4	Acetone	8	Type I	Type J
	temperature
6007235-19-E5	ambient	20	DMA	4	EtOAc	8	Type I	Type J
	temperature
6007235-19-E6	ambient	20	DMA	4	iPrOAc	8	Type I	Type J
	temperature
6007235-19-B6	ambient	25	DMSO	0.3	iPrOAc	3	—	Type J
	temperature

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 (see FIG. 25).

TABLE 8
					Solvent/
			Solvent/		Anti-
			Anti-	API	solvent	XRPD	XRPD
Exp.	Temp.	Temp	solvent	(mg)	(mL)	(wet cake)	(air-dry)
6007235-17-C6	Slow Cooling	50° C.-->4° C.	NMP	20.9	4	No solids
6007235-17-C14	Crash Cooling	50° C.-->−20° C.	NMP	26.2	4	Amorphous
6007235-17-C2	Slow Cooling	50° C.-->4° C.	CHCl3	25.4	4	Amorphous
						and A
6007235-17-C10	Crash Cooling	50° C.-->−20° C.	CHCl3	25.4	4	No solids
6007235-17-C13	Crash Cooling	50° C.-->−20° C.	DCM	22.1	10	No solids
6007235-17-C3	Slow Cooling	50° C.-->4° C.	EtOH	5	20	No solids
6007235-17-C11	Crash Cooling	50° C.-->−20° C.	EtOH	5	20	No solids
6007235-17-C9	Crash Cooling	50° C.-->−20° C.	MeOH	22.9	10	No solids
6007235-17-C1	Slow Cooling	50° C.-->4° C.	MeOH	24.7	20	No solids
6007235-17-C7	Slow Cooling	50° C.-->4° C.	DMA	25	4	Amorphous
						and A
6007235-17-C15	Crash Cooling	50° C.-->−20° C.	DMA	26.9	4	Amorphous
6007235-17-C8	Slow Cooling	50° C.-->4° C.	DMF	25.1	4	No solids
6007235-17-C16	Crash Cooling	50° C.-->−20° C.	DMF	25.1	4	Amorphous
6007235-17-C4	Slow Cooling	50° C.-->4° C.	ACN	12.6	10	Type C	Type C
6007235-17-C5	Slow Cooling	50° C.-->4° C.	DCM	26.8	10	Type C +	Type C
						peaks
6007235-17-C12	Crash cooling	50° C.-->−20° C.	ACN	12.6	10	Type C	Type 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 9
			API	Solvent
Exp	Temp.	Solvent	(mg)	(mL)	Crystal Type
6007235-17-B6	ambient temperature	methanol	12	10	Amorphous
6007235-17-B1	ambient temperature	methanol	12	10	Amorphous
6007235-17-B8	50° C.	ethanol	5	20	Amorphous and A
6007235-17-B3	50° C.	ethanol	5	20	Amorphous
6007235-17-B5	50° C.	dichloromethane	21.2	20	Amorphous
6007235-17-B10	50° C.	dichloromethane	28	20	Amorphous
6007235-17-B7	ambient temperature	CHCl3	27.4	7	Amorphous
6007235-17-B2	ambient temperature	CHCl3	26	7	Amorphous
6007235-17-B9	50° C.	acetonitrile	10.5	40	Amorphous
6007235-17-B4	50° C.	acetonitrile	10.5	40	Amorphous

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 (see FIG. 26). Anti-solvent studies in N,N-dimethylacetamide with methyl t-butyl ether yielded a HM30181 mesylate Type F polymorphism (see FIG. 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 (see FIG. 28). Other anti-solvent studies in DMSO and DMF yielded a HM30181 mesylate Type K polymorphism (or possibly a mixture of types, see FIG. 29). Anti-solvent experiments in DMF/n-propanol and DMA/isopropanol yielded a HM30181 mesylate Type L polymorphism (see FIG. 30). Anti-solvent addition DMF/Toluene and DMA/t-BuOH were mostly amorphous, but also generated HM30181 mesylate Type M (see FIG. 31).

Table 10 (continued on Table 11)
			Anti-
		Anti-	solvent		Evap
NB	Solvent	solvent	(mL)	Precipitation?	solids?	Identification
6004273-06-A1	DMSO	EtOH	20	N	Y	Type K
6004273-06-A2	DMSO	IPA	10	Y		insufficient
						solids/gel
6004273-06-A3	DMSO	Acetone	10	Y		Type K
6004273-06-A4	DMSO	MIBK	10	Y		Type K
6004273-06-A5	DMSO	EtOAc	10	Y		insufficient
						solids/gel
6004273-06-A6	DMSO	iPrOAc	10	Y		insufficient
						solids/gel
6004273-06-A7	DMSO	THF	10	Y		Similar to
						Type K
6004273-06-A8	DMSO	2-MeTHF	10	Y		Amorphous
6004273-06-A9	DMSO	1,4-Dioxane	20	Y		insufficient
						solids/gel
6004273-06-A10	DMSO	MTBE	20	Y		Amorphous
6004273-06-A11	DMSO	ACN	20	N	Y	Type C
6004273-06-A12	DMSO	DCM	20	N	Y	Similar to
						Type C
6004273-06-A13	DMSO	CHCl3	20	N	Y	Type K
6004273-06-A14	DMSO	Toluene	10	Y		Amorphous
6004273-06-A15	DMSO	Water	20	N	N	insufficient
						solids/gel
6004273-06-A16	DMSO	MEK	10	Y		insufficient
						solids/gel
6004273-06-A17	DMSO	t-Butanol	10	Y		Type K
6004273-06-A18	DMSO	n-Propanol	10	Y		Type K
6004273-06-A19	DMSO	n-Propyl	10	Y		Type K
		acetate
6004273-06-A20	CHCl3	Ethanol	10	Y		Type A
6004273-06-A21	CHCl3	IPA	10	Y		Type A
6004273-06-A22	CHCl3	Acetone	10	Y		Type A
6004273-06-A23	CHCl3	MIBK	10	Y		Type A
6004273-06-A24	CHCl3	EtOAc	10	Y		Type A
6004273-06-A25	CHCl3	iPrOAc	10	Y		Type A
6004273-06-A26	CHCl3	THF	10	Y		Type A
6004273-06-A27	CHCl3	2-MeTHF	10	Y		Type A
6004273-06-A28	CHCl3	1,4-Dioxane	10	Y		insufficient
						solids/gel
6004273-06-A29	CHCl3	MTBE	10	Y		Type A
6004273-06-A30	CHCl3	ACN	10	Y		Type A
6004273-06-A31	CHCl3	DCM	10	Y		Type A
6004273-06-A32	CHCl3	Toluene	10	Y		Type A
6004273-06-A33	CHCl3	n-Heptane	10	Y		Type A
Table 11 (continued from Table 10 and on Table 12)
			Anti-
			solvent		Evap
NB	Solvent	Anti-solvent	(mL)	Precipitation?	solids?	Identification
6004273-06-A34	CHCl3	MeOAc	10	Y		Type A
6004273-06-A35	CHCl3	Cyclohexane	10	Y		Type A
6004273-06-A36	CHCl3	MEK	10	Y		Type A
6004273-06-A37	CHCl3	t-Butanol	10	Y		Type A
6004273-06-A38	CHCl3	n-Propanol	10	Y		Type A
6004273-06-A39	CHCl3	n-Propyl	10	Y		Type A
		acetate
6004273-06-A40	CHCl3	Ethyl formate	10	Y		Type A
6004273-06-A41	CHCl3	iBuOAc	10	Y		Type A
6004273-06-A42	CHCl3	CPME	10	Y		Type A
6004273-06-A43	DMF	EtOH	20	N	Y	Amorphous
6004273-06-A44	DMF	IPA	10	Y		Type K
6004273-06-A45	DMF	Acetone	20	N	Y	insufficient
						solids/gel
6004273-06-A46	DMF	MIBK	10	Y		Amorphous
6004273-06-A47	DMF	EtOAc	10	Y		Type A
6004273-06-A48	DMF	iPrOAc	10	Y		insufficient
						solids/gel
6004273-06-A49	DMF	THF	20	N	Y	insufficient
						solids/gel
6004273-06-A50	DMF	2-MeTHF	10	Y		insufficient
						solids/gel
6004273-06-A51	DMF	1,4-Dioxane	20	N	Y	insufficient
						solids/gel
6004273-06-A52	DMF	MTBE	10	Y		Type A
6004273-06-A53	DMF	ACN	20	N	Y	insufficient
						solids/gel
6004273-06-A54	DMF	DCM	20	N	Y	insufficient
						solids/gel
6004273-06-A55	DMF	CHCl3	20	N	Y	insufficient
						solids/gel
6004273-06-A56	DMF	Toluene	10	Y		Type M
6004273-06-A57	DMF	Water	20	N	Y	insufficient
						solids/gel
6004273-06-A58	DMF	MEK	20	N	Y	insufficient
						solids/gel
6004273-06-A59	DMF	t-Butanol	10	Y		insufficient
						solids/gel
6004273-06-A60	DMF	n-Propanol	20	N	Y	Type L +
						amorphous
6004273-06-A61	DMF	n-Propyl	10	Y		Type A
		acetate
6004273-06-A62	NMP	EtOH	20	N	N	insufficient
						solids/gel
6004273-06-A63	NMP	IPA	10	Y		insufficient
						solids/gel
6004273-06-A64	NMP	Acetone	20	N	N	insufficient
						solids/gel
6004273-06-A65	NMP	MIBK	10	Y		insufficient
						solids/gel
6004273-06-A66	NMP	EtOAc	10	Y		insufficient
						solids/gel
Table 12 (continued from Table 11)
			Anti-
			solvent		Evap
NB	Solvent	Anti-solvent	(mL)	Precipitation?	solids?	Identification
6004273-06-A67	NMP	iPrOAc	10	Y		insufficient
						solids/gel
6004273-06-A68	NMP	THF	20	N	N	insufficient
						solids/gel
6004273-06-A69	NMP	2-MeTHF	10	Y		insufficient
						solids/gel
6004273-06-A70	NMP	1,4-Dioxane	20	N	N	Type A
6004273-06-A71	NMP	MTBE	10	Y		insufficient
						solids/gel
6004273-06-A72	NMP	ACN	20	N	N	insufficient
						solids/gel
6004273-06-A73	NMP	DCM	20	N	N	insufficient
						solids/gel
6004273-06-A74	NMP	CHCl3	20	N	N	insufficient
						solids/gel
6004273-06-A75	NMP	Toluene	10	Y		insufficient
						solids/gel
6004273-06-A76	NMP	Water	20	N	N	insufficient
						solids/gel
6004273-06-A77	NMP	MEK	20	N	Y	Amorphous
6004273-06-A78	NMP	t-BuOH	20	hazy	Y	Amorphous +
						Type J
6004273-06-A79	NMP	n-Propanol	20	N	N	insufficient
						solids/gel
6004273-06-A80	NMP	n-Propyl	10	Y		insufficient
		acetate				solids/gel
6004273-06-A81	DMA	EtOH	20	N	N	insufficient
						solids/gel
6004273-06-A82	DMA	IPA	20	hazy	Y	Type L +
						amorphous
6004273-06-A83	DMA	Acetone	20	N	Y	insufficient
						solids/gel
6004273-06-A84	DMA	MIBK	10	Y		Amorphous +
						Type J
6004273-06-A85	DMA	EtOAc	10	Y		insufficient
						solids/gel
6004273-06-A86	DMA	iPrOAc	10	Y		insufficient
						solids/gel
6004273-06-A87	DMA	THF	20	N	N	insufficient
						solids/gel
6004273-06-A88	DMA	2-MeTHF	10	Y		insufficient
						solids/gel
6004273-06-A89	DMA	1,4-Dioxane	20	N	N	insufficient
						solids/gel
6004273-06-A90	DMA	MTBE	10	Y		Amorphous +
						Type F
6004273-06-A91	DMA	ACN	20	N	N	insufficient
						solids/gel
6004273-06-A92	DMA	DCM	20	N	N	insufficient
						solids/gel
6004273-06-A93	DMA	CHCl3	20	N	N	insufficient
						solids/gel
6004273-06-A94	DMA	Toluene	10	Y		Amorphous +
						Type J
6004273-06-A95	DMA	Water	20	N	Y	insufficient
						solids/gel
6004273-06-A96	DMA	MEK	20	N	N	insufficient
						solids/gel
6004273-06-A97	DMA	t-Butanol	20	N	Y	Type M
6004273-06-A98	DMA	n-Propanol	20	N	Y	insufficient
						solids/gel
6004273-06-A99	DMA	n-Propyl	10	Y		insufficient
		acetate				solids/gel

TABLE 13
			Anti-
			solvent		Evap
NB	Solvent	Anti-solvent	(mL)	Precipitation?	solids?	Identification
6004273-06-A100	NMP	1,4-Dioxane	20	N		insufficient
						solids/gel
6004273-06-A101	NMP	MIBK	10	Y		insufficient
						solids/gel
6004273-06-A102	CHCl3	EtOH	10	Y		Type A
6004273-06-A103	CHCl3	MTBE	10	Y		Type A
6004273-06-A104	CHCl3	n-Heptane	10	Y		Type A
6004273-06-A105	DMSO	EtOAc	10	Y		insufficient
						solids/gel
6004273-06-A106	DMSO	Toluene	10	Y		Amorphous
6004273-06-A107	DMSO	Water	20	N		insufficient
						solids/gel
6004273-06-A108	DMA	Acetone	20	N		insufficient
						solids/gel
6004273-06-A109	DMA	iPrOAc	10	Y		insufficient
						solids/gel
6004273-06-A110	DMF	IPA	10	Y		Amorphous +
						Type K
6004273-06-A111	DMF	THF	20	N		insufficient
						solids/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 (see FIG. 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 (see FIG. 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 (see FIG. 33). A scaled-up slurry of HM30181 mesylate Type A starting material in NMP provided a primarily amorphous material (see FIG. 34). A scaled-up slurry of HM30181 mesylate Type A starting material in DMA yielded HM30181 mesylate Type E after 6 days (see FIG. 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 (see FIG. 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 (see FIG. 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 (see FIG. 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 (see FIG. 37). 1H-NMR results in DMSO-d₆ suggested that the DMF slurry at 150° C. resulted in degradation, as the 1H-NMR spectrum did not match either the starting material or freebase (see FIG. 38).

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

TABLE 14
						Anti-
			Solvent	Conc.	Anti-	solvent	Desired
NB	Method	Solvent	(mL)	(mg/mL)	solvent	(mL)	Type	Observations
6004273-10-B	Slurry at	MeOH	0.2	480			B	Type N
	ambient
	temperature
6004273-10-C	Slurry at	MeCN	0.2	545			C	Type C
	ambient
	temperature
6004273-10-D	Slurry at	NMP	0.2	550			D	Amorphous
	ambient
	temperature
6004273-10-E	Slurry at	DMA	0.2	576			E	Type E
	ambient
	temperature
6004273-10-F	Slurry at	DMF	0.2	595			F	Type A
	ambient
	temperature
6004273-10-	Slurry at	DMF	0.2	595			G	Type F
F-100C	100° C.
	(for 2 days)
6004273-10-	Slurry at	DMF	0.2	595			G	Decomposition
F-150C	150° C.
	(for 5 d)
6004273-10-G	Slurry at	DMF	0.2	576			G	Type F
	50° C.
6004273-10-H	Liquid	DMSO	5	60	MeCN	3	H	No precipitation,
	vapor							evaporated- no
	sorption							solids due to
								DMSO
6004273-10-J	Liquid	DMSO	10	60	Acetone	3	J	No precipitation,
	vapor							evaporated- no
	sorption							solids due to
								DMSO
6004273-10-K	Anti-	DMSO	10	60	MIBK	200	K	Amorphous
	solvent
	addition
6004273-10-L	Anti-	DMF	20	5	n-propanol	200	L	Amorphous
	solvent
	addition
6004273-10-M	Anti-	DMA	20	5	t-BuOH	200	M	Amorphous
	solvent
	addition
6004273-10-M1	Anti-	DMF	20	5	Toluene	300	M	Amorphous
	solvent
	addition

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 (see FIG. 40). By DSC, HM30181 mesylate Type B displayed an endotherm at 159.92° C. (see FIG. 41). By TGA, HM30181 mesylate Type B showed 1.865% weight loss before 170° C., followed by possible disassociation and decomposition (see FIG. 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 (see FIG. 42). By DSC, HM30181 mesylate Type C displays an endotherm at 159.60° C. (see FIG. 43). By TGA, HM30181 mesylate Type C showed 2.987% weight loss before 170° C., followed by a disassociation and decomposition (see FIG. 43). By 1H-NMR, HM30181 mesylate Type C is confirmed to contain only water and no acetonitrile, (˜2.07 ppm) suggesting a monohydrate (see FIG. 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 (see FIG. 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 (see FIG. 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. (see FIG. 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 (see FIG. 48). By ¹H-NMR, HM30181 mesylate Type E was confirmed to contain DMA (see FIG. 49).
  • Crystalline HM30181 mesylate Type F was obtained through slurrying in dimethylformamide at 50° C. and shows characteristic results by XRPD (see FIG. 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 (see FIG. 51). By ¹H-NMR, HM30181 mesylate Type F was confirmed to contain DMF (see FIG. 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 (see FIG. 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 (see FIG. 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 (see FIG. 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 (see FIG. 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 (see FIG. 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 (see FIG. 58). By TGA, HM30181 mesylate Type J displayed a 2.887% weight loss before 150° C., suggesting a solvate or hydrate (see FIG. 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 (see FIG. 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 (see FIG. 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 (see FIG. 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 (see FIG. 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 (see FIG. 64). 1H-NMR confirmed the presence of methanol (see FIG. 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 15
		FWHM		Rel.
Pos.	Height	Left	d-spacing	Int.
[°2Th.]	[cts]	[°2Th.]	[Å]	[%]
3.5	34.6	0.05	25.5	1.8
5.1	95.5	0.115	17.5	5.0
5.6	30.2	0.08	15.8	1.6
6.4	419.6	0.08	13.7	22.2
8.0	1247.3	0.12	11.0	65.9
9.9	37.7	0.20	8.9	2.0
11.7	305.9	0.20	7.6	16.2
12.1	322.7	0.10	7.3	17.1
12.8	1892.0	0.10	6.9	100.0
13.9	205.8	0.13	6.4	10.9
15.0	609.3	0.13	5.9	32.2
15.5	113.8	0.82	5.7	6.0
16.1	253.9	0.15	5.5	13.4
17.8	71.2	0.46	5.0	3.8
19.2	439.5	0.20	4.6	23.2
19.7	590.3	0.20	4.5	31.2
20.0	362.7	0.20	4.4	19.2
21.2	60.7	0.10	4.2	3.2
22.1	20.0	0.10	4.0	1.0
22.9	309.0	0.13	3.9	16.3
23.4	99.7	0.18	3.8	5.3
24.5	37.2	0.20	3.6	2.0
25.1	63.6	0.31	3.6	3.4
26.1	1066.2	0.31	3.4	56.4
26.9	130.0	0.20	3.3	6.9
28.1	144.2	0.23	3.2	7.6
29.4	15.4	0.31	3.0	0.8
32.4	19.0	0.20	2.8	1.0
34.3	25.9	0.05	2.6	1.4
35.50	13.2	0.41	2.5	0.7
36.5	16.1	0.15	2.5	0.9
37.3	12.0	0.08	2.4	0.6

TABLE 16
Reflection Conditions
Crystal System         Triclinic
Bravais Type           Primitive (P)
Space Group
Instrument Settings
Goniometer Radius (mm) 240.00
Unit Cell
a (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)            1179.70
Refinement Results
No. Unindexed Lines    0
No. Indexed Lines      17
Total No.              5110
Calculated Lines
Chi Square             3.777537E−0006
Snyder's FOM           2.0791

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 17
		FWHM		Rel.
Pos.	Height	Left	d-spacing	Int.
[°2Th.]	[cts]	[°2Th.]	[Å]	[%]
4.2	458.0	0.15	21.2	43.7
5.2	4.1	0.05	17.0	0.4
6.0	16.0	0.038	14.7	1.5
6.2	13.5	0.051	14.3	1.3
6.3	22.1	0.038	14.0	2.1
8.2	1047.3	0.15	10.8	100.0
9.0	35.8	0.038	9.9	3.4
9.1	29.7	0.15	9.7	2.8
9.9	13.9	0.05	9.0	1.3
10.4	344.2	0.13	8.5	32.9
10.7	430.1	0.15	8.3	41.1
11.6	136.8	0.20	7.6	13.1
12.3	114.7	0.18	7.2	11.0
12.9	170.3	0.15	6.9	16.3
13.6	30.8	0.15	6.5	2.9
14.7	685.3	0.26	6.0	65.4
15.4	227.7	0.13	5.7	21.8
15.9	801.8	0.23	5.6	76.6
16.3	46.5	0.15	5.4	4.4
16.8	827.7	0.13	5.3	79.0
17.7	281.7	0.20	5.0	26.9
18.3	200.1	0.23	4.9	19.1
18.9	298.0	0.20	4.7	28.4
19.3	164.2	0.13	4.6	15.7
19.6	106.5	0.13	4.5	10.2
20.0	103.1	0.15	4.4	9.8
20.4	153.2	0.13	4.3	14.6
21.0	842.3	0.22	4.2	80.4
21.3	320.9	0.13	4.2	30.6
21.6	243.7	0.10	4.1	23.3
22.0	111.1	0.20	4.0	10.6
22.4	128.7	0.15	4.0	12.3
22.7	418.2	0.26	3.9	39.9
23.8	272.6	0.26	3.7	26.0
24.1	116.9	0.10	3.7	11.2
24.6	174.5	0.20	3.6	16.7
24.9	214.9	0.18	3.6	20.5
26.2	994.9	0.15	3.4	95.0
26.6	965.7	0.15	3.3	92.2
27.7	343.3	0.20	3.2	32.8
28.3	94.3	0.18	3.2	9.0
28.8	20.2	0.15	3.1	1.9
29.3	62.1	0.15	3.0	5.9
30.7	45.9	0.20	2.9	4.4
32.1	117.5	0.18	2.8	11.2
33.3	40.2	0.15	2.7	3.8
33.9	42.7	0.31	2.6	4.1
34.7	33.4	0.26	2.6	3.2
35.7	22.6	0.20	2.5	2.2
37.0	29.8	0.15	2.4	2.8

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 18
	Reflection Conditions
	Crystal System	Triclinic
	Bravais Type	Primitive (P)
	Space Group
	Instrument Settings
	Goniometer Radius (mm)	240.00
	Unit Cell
	a (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)	1618.45
	Refinement Results
	No. Unindexed Lines	0
	No. Indexed Lines	32
	Total No.	6088
	Calculated Lines
	Chi Square	8.802423E−0007
	Snyder's FOM	5.7186

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 19
Abbreviation	Solvent	Abbreviation	Solvent
MeOH	Methanol	THF	Tetrahydrofuran
EtOH	Ethanol	2-MeTHF	2-Methyltetrahydrofuran
IPA	Isopropyl	DMF	Dimethyl formamide
	alcohol
ACN	Acetonitrile	DMSO	Dimethyl sulfoxide
MIBK	Methyl isobutyl	CHCl3	Chloroform
	ketone
EtOAc	Ethyl acetate	DCM	Dichloromethane
iPrOAc	Isopropyl	DMAc	Dimethylacetamide
	acetate
MTBE	Methyl tert-butyl	t-BuOH	t-Butanol
	ether

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 20
Parameters for Reflection Mode
X-Ray wavelength   Cu, kα, Kα1 (Å): 1.540598,
                   Kα2 (Å): 1.544426
                   Kα2/Kα1 intensity ratio: 0.50
X-Ray tube setting 45 kV, 40 mA
Divergence slit    Automatic
Scan mode          Continuous
Scan range (°2TH)  3°-40°
Step size (°2TH)   0.0131
Scan 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 N₂ as the purge gas, with pan crimped (see Table 21).

TABLE 21
Parameters  DSC
Pan Type    Aluminum pan, closed
Temperature ambient temperature-300° C.
Ramp rate   10° C./min
Purge gas   N2

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 N₂ as the purge gas, then cooled by 10° C. to 25° C. This temperature cycle repeated twice (see Table 22).

TABLE 22
	Parameters	DSC
	Pan Type	Aluminum pan, closed
	Temperature	25-150°	C.
	Ramp rate	10°	C./min
	Purge gas	N2
	Heat/cool Cycles	2

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 N₂ as the purge gas, with pan open (see Table 23).

TABLE 23
	Parameters	TGA
	Pan Type	Platinum plate, open
	Temperature	300°	C.
	Ramp rate	10°	C./min
	Purge gas	N2
	Sample purge flow	15	mL/min
	Balance purge flow	25	mL/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 24
Parameters	Values
Temperature	25°	C.
Sample size	10-20	mg
Gas and flow rate	N2, 200 mL/min
dm/dt	0.002%/min
Min. dm/dt stability duration	10	min
Max. equilibrium time	360	min
RH range	40% RH-95% RH-0% RH-95% RH
RH step size	10%	(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.

Claims

1. A composition comprising a crystalline form of HM30181 mesylate, where the crystalline form is polymorph B, and wherein the crystalline form has an X-ray diffraction pattern corresponding to FIG. 40 and has an endotherm at about 159.92° C.

2. A method of inhibiting P-glycoprotein activity, comprising contacting P-glycoprotein with a compound of claim 1 in an amount effective to inhibit an activity of P-glycoprotein.

3. A method of treating cancer, comprising: administering a chemotherapeutic drug to an individual in need of treatment; andadministering the crystalline form of HM30181 mesylate of claim 1 to the individual in need of treatment in an amount effective to inhibit P-glycoprotein activity in the individual,wherein the chemotherapeutic drug is a P-glycoprotein substrate.