# Patent

## Title
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 polymorphism C, as well as their physical properties and methods for their preparation and characterization.

## Background
This application claims the benefit of U.S. Provisional Patent Application No. 63/107,720 filed on Oct. 30, 2020, and Provisional Patent Application No. 63/107,792 filed on Oct. 30, 2020. These and all other referenced extrinsic materials are incorporated herein by reference in their entirety. Where a definition or use of a term in a reference that is incorporated by reference is inconsistent or contrary to the definition of that term provided herein, the definition of that term provided herein is deemed to be controlling.


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 Å3, 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 Å3, 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









Table 1

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 4 (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.
 
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 (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
 
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


 
 
 
 
 


 
 
 
 
 


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]
[%]







 
 
 
 
 


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


 
 
 
 
 


 
 
 
 
 


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


 
 
 
 
 


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


 
 
 
 
 


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


 
 
 
 
 


21.3
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


 
 
 
 
 


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


  
 
 
 
 


 
 
 
 
 


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'Pert3TM Powder XRPD and on a Si zero-background holder. The 2θ position was calibrated against a PanalyticalTM 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 Q2000TM 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 Q500TM 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 SystemsTM) 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 200MTM 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 wherein the crystalline form is polymorph C, and wherein the crystalline form has an X-ray diffraction pattern corresponding to FIG. 42 and has an endotherm at about 159.6° C.

2. A method of inhibiting P-glycoprotein activity, comprising contacting P-glycoprotein with the crystalline form of HM30181 mesylate of claim 1 in an amount effective to inhibit an activity of P-glycoprotein.