18-MC SALT FORMS

A composition including a salt of 18-MC, wherein the salt is chosen from gentisate, hydrobromide, besylate, napadisylate, hydrochloride, sulfate, oxalate, maleate, mesylate, and tosylate. A composition including a polymorph of 18-MC.

BACKGROUND OF THE INVENTION

1. Technical Field

The present invention relates to compositions of salts and polymorphs of 18-methoxycoronaridine.

2. Background Art

18-methoxycoronaridine (18-MC) is a derivative of ibogaine with the chemical formula of C22H28N2O3. The freebase 18-MC is a synthetic coronaridine congener and a specific negative allosteric modulator (antagonist) of α3β4 nicotinic cholinergic receptors; it indirectly modulates the dopaminergic mesolimbic pathway via blockade of α3β4 nicotinic receptors in the habenulo-interpeduncular pathway and the basolateral amygdala (Glick et al., 2008). Animal studies have demonstrated that 18-MC significantly reduces drug self-administration in a number of substance use models (nicotine, alcohol, morphine, cocaine and methamphetamine) at dosages as low as 10 mg/kg i.p. (Glick et al., 1994; Rezvani et al., 1995; Glick et al., 1996; Glick et al., 1998; Glick et al., 2000a). More recently, 18-MC has been shown in an animal model to attenuate effects of the environmental cues responsible for stimulating cocaine-seeking or “craving” behaviors (Polston et al., 2012, and U.S. Pat. Application No. 14/387,339 to Glick, et al.). This property of 18-MC could potentially help address the craving component of human addictive behaviors.

U.S. Pat. Application No. 14/387,339 to Glick, et al. discloses methods of preventing drug relapse, especially during cue inducement, by administering an effective amount of an α3β4 nicotinic antagonist (18- Methoxycoronaridine) to a mammal, after an initial period of drug use, and preventing a relapse of drug use. It was shown that rats conditioned with a musical cue show increased drug-seeking behaviors with cocaine when compared to control groups. Pharmaceutically acceptable HCl salts of 18-MC are mentioned, however, polymorphism is not disclosed. Preparation of the 18-MC HCl salt has been briefly described in the literature (Acta Crystallographica Section E, Structure Reports, ISSN 1600-5368, Acta Cryst. (2012). E68, o1041). However, few details are provided and conflicting information is provided regarding the solvent system. Understanding and control of polymorphism is required in order to develop a robust and scalable API manufacturing process resulting in a stable material that is suitable for drug product manufacturing.

Therefore, there remains a need for salts and polymorphs of 18-MC.

SUMMARY OF THE INVENTION

The present invention provides for a composition including a salt of 18-MC, wherein the salt is chosen from gentisate, hydrobromide, besylate, napadisylate, hydrochloride, sulfate, oxalate, maleate, mesylate, and tosylate.

The present invention provides for a composition including a polymorph of 18-MC, wherein the polymorph is chosen from HCl salt Type A, HCl salt Type B, HCl salt Type C, HCl salt Type D, HCl salt Type E, HCl salt Type F, HCl salt Type G, HCl salt Type H, HCl salt Type I, HCl salt Type J, HCl salt Type K, HCl salt Type L, HCl salt Type M, HCl salt Type N, HCl salt Type O, HCl salt Type P, HCl salt Type Q, HCl salt Type R, HCl salt Type S, HCl salt Type T, HCl salt Type U, HCl salt Type V, sulfate salt Type A, sulfate salt Type B, sulfate salt Type C, sulfate salt Type D, sulfate salt Type E, sulfate salt Type F, oxalate salt Type A, oxalate salt Type B, maleate salt Type A, mesylate salt Type A, mesylate salt Type B, mesylate salt Type C, HBr salt Type A, HBr salt Type B, HBr salt Type C, HBr salt Type D, tosylate salt Type A, tosylate salt Type B, tosylate salt Type C, tosylate salt Type D, tosylate salt Type E, tosylate salt Type F, tosylate salt Type G, tosylate salt Type H, tosylate salt Type I, besylate salt Type A, besylate salt Type B, besylate salt Type C, napadisylate salt Type A, napadisylate salt Type B, napadisylate salt Type C, napadisylate salt Type D, and gentisate salt Type A.

DETAILED DESCRIPTION OF THE INVENTION

The present invention provides for salts and polymorphs of 18-MC. The polymorphs can be crystalline or amorphous.

The polymorphs can include HCl salt Type A, HCl salt Type B, HCl salt Type C, HCl salt Type D, HCl salt Type E, HCl salt Type F, HCl salt Type G, HCl salt Type H, HCl salt Type I, HCl salt Type J, HCl salt Type K, HCl salt Type L, HCl salt Type M, HCl salt Type N, HCl salt Type O, HCl salt Type P, HCl salt Type Q, HCl salt Type R, HCl salt Type S, HCl salt Type T, HCl salt Type U, HCl salt Type V, sulfate salt Type A, sulfate salt Type B, sulfate salt Type C, sulfate salt Type D, sulfate salt Type E, sulfate salt Type F, oxalate salt Type A, oxalate salt Type B, maleate salt Type A, mesylate salt Type A, mesylate salt Type B, mesylate salt Type C, HBr salt Type A, HBr salt Type B, HBr salt Type C, HBr salt Type D, tosylate salt Type A, tosylate salt Type B, tosylate salt Type C, tosylate salt Type D, tosylate salt Type E, tosylate salt Type F, tosylate salt Type G, tosylate salt Type H, tosylate salt Type I, besylate salt Type A, besylate salt Type B, besylate salt Type C, napadisylate salt Type A, napadisylate salt Type B, napadisylate salt Type C, napadisylate salt Type D, and gentisate salt Type A.

Crystalline gentisate salt Type A can be characterized by an x-ray powder diffraction pattern having peaks expressed as 20 at about 10.3, about 11.1, about 16.3, about 20.6, about 21.0, and about 27.8. Crystalline HBr salt Type A can be characterized by an x-ray powder diffraction pattern having peaks expressed as 20 at about 8.6, about 13.1, about 19.1, about 19.9, about 26.1, and about 26.3. Crystalline HBr salt Type B can be characterized by an x-ray powder diffraction pattern having peaks expressed as 20 at about 7.5, about 15.0, about 21.2, about 21.9, about 24.1, and about 30.3. Crystalline besylate salt Type A can be characterized by an x-ray powder diffraction pattern having peaks expressed as 20 at about 7.4, about 8.1, about 14.3, about 14.7, about 19.7, and about 22.7. Crystalline besylate salt Type B can be characterized by an x-ray powder diffraction pattern having peaks expressed as 20 at about 8.3, about 9.8, about 16.6, about 17.7, about 18.4, and about 18.7. Crystalline napadisylate salt Type A can be characterized by an x-ray powder diffraction pattern having peaks expressed as 20 at about 7.6, about 8.1, about 12.2, about 12.7, about 14.6, and about 17.5.

Crystalline napadisylate salt Type B can be characterized by an x-ray powder diffraction pattern having peaks expressed as 20 at about 8.2, about 10.6, about 17.8, about 19.3, about 20.0, and about 21.3. Crystalline napadisylate salt Type C can be characterized by an x-ray powder diffraction pattern having peaks expressed as 20 at about 7.3, about 9.6, about 15.2, about 18.4, about 19.1, and about 24.5. Crystalline napadisylate salt Type D can be characterized by an x-ray powder diffraction pattern having peaks expressed as 20 at about 7.3, about 7.5, about 15.0, about 17.8, about 18.0, and about 22.5. Crystalline HCl salt Type A and is characterized by an x-ray powder diffraction pattern having peaks expressed as 20 at about 8.8, about 11.0, about 13.4, about 16.2, about 16.5, and about 16.8. Crystalline sulfate salt Type A can be characterized by an x-ray powder diffraction pattern having peaks expressed as 20 at about 5.2, about 10.5, about 13.8, about 15.7, about 18.3, and about 20.4.

Crystalline maleate salt Type A can be characterized by an x-ray powder diffraction pattern having peaks expressed as 20 at about 7.9, about 14.3, about 14.7, about 15.9, about 18.3, and about 19.1. Crystalline tosylate salt Type A can be characterized by an x-ray powder diffraction pattern having peaks expressed as 20 at about 8.6, about 10.4, about 11.8, about 17.9, about 18.2, and about 21.0. Crystalline tosylate salt Type B can be characterized by an x-ray powder diffraction pattern having peaks expressed as 20 at about 7.4, about 7.6, about 9.6, about 11.6, about 14.9, and about 15.3. Crystalline mesylate salt Type A can be characterized by an x-ray powder diffraction pattern having peaks expressed as 20 at about 8.1, about 9.2, about 13.0, about 16.9, about 18.2, and about 21.1. Crystalline oxalate salt Type A can be characterized by an x-ray powder diffraction pattern having peaks expressed as 20 at about 6.0, about 9.1, about 13.6, about 15.8, about 18.2, and about 21.8. Crystalline oxalate salt Type B can be characterized by an x-ray powder diffraction pattern having peaks expressed as 20 at about 7.7, about 11.8, about 13.7, about 16.7, about 17.7, and about 18.9. The freebase can be characterized by an x-ray powder diffraction pattern having peaks expressed as 20 at about 11.0, about 11.7, about 14.0, about 15.5, about 18.3, and about 21.3.

A formal, broad salt and polymorph screen was conducted resulting in at least 10 pharmaceutically relevant salts. Most of these exhibit polymorphism that was further characterized as described in the EXAMPLES below. The hydrochloride salt exhibits at least 22 different forms. Form A is the most thermodynamically stable form. Unexpectedly, however, Form J was isolated on up to ~100 g scale when isolated from HCl/EtOAc. Thus, appropriate control of the manufacturing process is critical to obtain the desired salt and polymorph.

The current standard process for making 18-MC (isolation from dioxane/HCI) ensures the controlled isolation of Form A.

Example 1 - Salt Screening of 18-MC

Applicant performed an extended salt screening and collected data of alternative salts.

18-MC Freebase was first isolated from 18-MC HCl salt and then used for salt screening. In the screening, a total of 100 experiments were performed using 20 acids and 5 solvents. Resulting solids were characterized by X-ray powder diffraction (XRPD). Based on XRPD comparison results, new forms were then characterized by thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), proton nuclear magnetic resonance (1H NMR) high-performance liquid chromatography (HPLC) and/or ion chromatography (IC). As the results showed, a total of 16 salt forms were discovered from screening and form re-preparation, including gentisate Type A, HBr salt Types A and B, besylate Types A and B and napadisylate Types A, B, C′ and D. XRPD patterns are shown inFIG.1and characterization results are summarized in TABLE 1A. Additional Salt screening results are summarized in TABLE 1B. A summary of all salts and salt forms produced during the study are in TABLE 5B.

To summarize, a total of 16 new salt forms were found and fully characterized in the extended salt screening.

Freebase Isolation

Characterization of Starting Material

Three batches of starting material of 18-MC HCl salt were characterized by XRPD, TGA, DSC and HPLC/IC. Characterization results are summarized in TABLE 2A and displayed fromFIGS.2A to21. As the characterization results showed, the three batches represent different HCl salt polymorphs.

As shown inFIG.2Aand TABLE 2B, starting material (824509-01-A) was crystalline and conformed to HCl salt Type A. TGA/DSC results inFIG.2Bshowed that a weight loss of 0.8% up to 150° C. and one endothermic signal at 207.8° C. (onset) was observed before decomposition. The Cl- content of the material was determined as 8.73% (theoretical Cl- content for mono HCl salt is 8.77%), and HPLC purity was 99.33 area% (FIG.2Cand TABLE 2C).

As shown inFIG.2D, the sample (824509-20-A,) was amorphous. TGA result inFIG.2Eshowed a weight loss of 7.7% up to 120° C. HPLC/IC results showed that the molar ratio of hydrochloric acid to 18-MC freebase was determined as 1.1:1 and HPLC purity was 97.85 area% (FIG.2Fand TABLE 2D).

TABLE 2DHPLC results of starting material (824509-20-A)#RRTArea(%)#RRTArea(%)10.730.1870.960.5120.880.2980.980.2930.900.0791.0097.8540.910.05101.070.2350.930.12111.080.0560.940.28121.100.09

As shown inFIG.2G, the sample (824509-20-B) was similar to HCl salt Type H, with extra peaks similar to HCl salt Type A and B (marked in red frame). TGA/DSC curves inFIG.2Hshowed a weight loss of 4.2% up to 130.0° C. and three endothermic signals at 94.5° C., 163.8° C. and 190.9° C. (peak temperature). HPLC/IC results showed that the molar ratio of acid/freebase was determined as 1.1:1 and HPLC purity was 97.02 area% (FIG.21and TABLE 2E).

TABLE 2EHPLC results of starting material (824509-20-B)#RRTArea (%)#RRTArea (%)10.730.2990.980.4820.870.05101.0097.0230.880.24111.070.2640.900.08121.080.0650.910.08131.100.1360.930.12141.110.1270.940.37151.140.0680.960.63------

Freebase Isolation

A detailed procedure for the preparation of 18-MC Freebase from 18-MC HCl Salt can be found below, and characterization results of prepared freebase batches are summarized in TABLE 2F. Additional data are shown inFIGS.2J to2Pand TABLE 2F to 21.

For freebase Type A (824509-03-A), the Cl- contents was determined to be less than 0.24%. TGA/DSC curves fromFIG.2Kshowed that up to 150° C., a TGA weight loss of 1.1 % was observed and one endothermic signal around 196.2° C. (onset) was detected. HPLC chromatograms fromFIG.2Ldisplayed the HPLC purity was around 99.31 area% (TABLE 2G).

TABLE 2GHPLC results of freebase Type A (824509-03-A)#RRTArea(%)#RRTArea(%)10.920.1541.070.2720.970.0951.270.0531.0099.3161.310.13

For freebase Type A (824509-21-A), the Cl- contents was determined to be less than 0.15%. TGA/DSC curves fromFIG.2Mshowed that up to 180° C., a TGA weight loss of 1.1 % was observed and one endothermic signal around 196.7° C. (onset) were detected. HPLC chromatograms fromFIG.2Ndisplayed that the HPLC purity was determined to be 99.45 area% (TABLE 2H).

TABLE 2HHPLC results of freebase Type A (824509-21-A)#RRTArea (%)#RRTArea (%)10.730.0541.0099.4720.930.0951.070.1430.960.25------

For freebase Type A (824509-24-A), no Cl- residual in the sample was detected. TGA/DSC curves fromFIGS.20showed that up to 170° C., a TGA weight loss of 1.9% was observed and one endothermic signal around 192.7° C. (onset) was detected. HPLC chromatograms fromFIG.2Pdisplayed that the HPLC purity was determined to be 98.23 area% (TABLE 21).

TABLE 2IHPLC results of freebase Type A (824509-24-A)#RRTArea (%)#RRTArea (%)10.730.2670.960.4120.880.0780.980.3730.900.0791.0098.2340.910.06101.070.1550.930.11111.100.0560.940.23------

Salt Screening

Using prepared freebase Type A (batch 824509-03-A and 824509-24-A) as starting material, a total of 100 experiments of salt screening were conducted with 20 acids and different solvent systems. For the CHCl3system, a stock solution of freebase was first prepared by dissolving ~150 mg of freebase Type A sample in 3.75 mL of CHCl3. Then the corresponding acids (charge molar ratio of acid/freebase=1:1) were added in 0.5 mL of stock solutions and slurried at RT. For the remaining solvent systems (IPAc, MIBK, 1,4-dioxane, IPA, DCM/EtOAc (1:1, v/v), acetone/H20 (9:1, v/v) and ACN/THF (3:1, v/v)), about 20 mg of freebase Type A sample was weighed into each HPLC vial and mixed with corresponding solid acids (charge molar ratio of acid/freebase=1:1). About 0.5 mL of solvents were added into the vial and the mixture was transferred to slurry at RT. Liquid acids were first diluted with 0.25 mL solvent and then added into freebase solution (freebase in 0.25 mL solvent). After slurry for about 5 days, precipitates were separated by centrifugation and the resulting solids were vacuum-dried at RT overnight (4 to 15 hours). If oil or gel-like samples were obtained after slurry, the samples were transferred to temperature cycling (one cycle: ramp to 50° C. at a rate of 4.5° C./min, keep the temperature at 50° C. for 30 min; cool down to 5° C. at a rate of 0.1° C./min and keep the temperature at 5° C. for 30 min. 4 cycles were conducted). If clear solution was obtained, it was transferred to slurry at 5° C. If it was still clear, the solution was transferred to slurry at -20° C. If there was still no precipitate, the clear solution was transferred to evaporation at RT or anti-solvent addition to induce precipitation. After vacuum drying at RT, all the resulting dry solids were tested by XRPD.

As the XRPD comparison results in TABLE 3A and TABLE 3B shows, a total of 9 salt forms were discovered, including gentisate Type A, HBr salt Types A and B, besylate Types A and B and napadisylate Types A, B, C, and D. XRPD overlay was displayed inFIG.1. The characterization data of these salt hits was summarized in TABLE 1A.

Characterization of Salt Hits

Gentisate Type A (824511-01-C9) was obtained by stirring 8.3 mg gentisic acid in 0.5 mL stock solution of freebase (40 mg/mL, charge molar ratio of acid to freebase was 1:1) in CHCl3at RT for about 5 days. Resulting solids were isolated by centrifugation and vacuum dried at RT overnight. The XRPD pattern was displayed inFIG.3Aand TABLE 4A. TGA/DSC curves inFIG.3Bshowed that a weight loss of 2.7% up to 150° C. and one endothermic signal at 181.9° C. (peak) were detected.1H NMR spectrum inFIG.3Cshowed that peaks of gentisic acid and CHCl3were observed. The molar ratio of acid/freebase was 1.0:1. The molar ratio of CHCl3/API was 0.05:1 (theoretical weight=1.1 wt%). HPLC purity of the sample was determined as 99.79 area% (FIG.3Dand TABLE 4B).

TABLE 4BHPLC results of gentisate Type A (824511-01-C9)#RRTArea (%)10.970.0621.0099.7931.070.15

HBr Salt

HBr Salt Type A

HBr salt Type A (824511-01-E10) was obtained by diluting 11.0 µL HBr (~40% aqueous solution) in 0.25 mL IPA and suspending 19.8 mg freebase in 0.25 mL IPA at RT, then adding acid solution to freebase suspension (charge molar ratio of acid to freebase was 1:1) and slurry at RT for about 5 days. Resulting solids were isolated by centrifugation and vacuum dried at RT overnight. The XRPD pattern was displayed inFIG.3Eand TABLE 4C. As TGA/DSC curves inFIG.3Fshown, a weight loss of 1.5% up to 150° C. and one endothermic signal at 208.5° C. (peak) were detected.1H NMR spectrum inFIG.3Gshowed that peak of IPA was observed. The molar ratio of IPA/API was 0.05:1 (theoretical 0.6 wt%). HPLC/IC results showed that the molar ratio of acid/freebase was 1.0:1 and HPLC purity was 99.72 area% (FIG.3Hand TABLE 4D).

TABLE 4DHPLC results of HBr salt Type A (824511-01-E10)#RRTArea (%)10.970.0621.0099.7231.070.23

HBr Salt Type B

HBr salt Type B (824511-01-D10) was obtained by diluting 11.0 µL HBr (~40% aqueous solution) in 0.25 mL 1,4-dioxane and suspending 20.1 mg freebase in 0.25 mL 1,4-dioxane at RT, then adding acid solution to freebase suspension (charge molar ratio of acid to freebase was 1:1) and slurry at RT for about one week. Resulting solids were isolated by centrifugation and vacuum dried at RT overnight. Another batch of HBr salt Type B (824511-10-A1) was prepared using the same method, and the XRPD overlay was displayed inFIG.3Iwith XRPD peak list shown in TABLE 4E. As TGA/DSC curves inFIG.3Jshowed, a weight loss of 14.2% up to 150° C., two endothermic signals at 104.3° C. (peak), 140.3° C. (peak) and one exothermic signal at 177.4° C. (peak) were detected.1H NMR spectrum inFIG.3Kshowed that a peak of 1,4-dioxane was observed. The molar ratio of 1,4-dioxane/API was 0.6:1 (theoretical 10.8 wt%). HPLC/IC results showed that the molar ratio of acid/freebase was determined as 0.9:1 and HPLC purity was 99.27 area% (FIG.3Land TABLE 4F).

TABLE 4FHPLC results of HBr salt Type B (824511-10-A1)#RRTArea(%)#RRTArea(%)10.750.0950.950.0620.830.0560.970.1330.900.1371.0099.2740.930.0981.070.17

Napadisylate Type A

Napadisylate Type A (824511-30-B7) was obtained by slurry ~20.0 mg freebase and 16.2 mg naphthalene-1,5-disulfonic acid (charge molar ratio of acid to freebase was 1:1) in 0.5 mL acetone/H2O (9:1, v/v) at RT for about one week. Resulting solids were isolated by centrifugation and vacuum drying at RT overnight. The XRPD pattern was displayed inFIG.3Mand TABLE 4G. As TGA/DSC curves inFIG.3Nshowed, a weight loss of 3.5% up to 70° C., 3.6% from 70° C. up to 120° C. and three endothermic signals at 96.6° C., 163.0° C. and 198.5° C. (peak) were observed.1H NMR spectrum in FIGURE showed that the peak of naphthalene-I,5-disulfonic acid and acetone were observed. The molar ratio of acid/API was 0.6:1, the molar ratio of acetone/API was 0.02:1 (theoretical 0.26 wt%). HPLC purity was 99.21 area% (FIG.3Pand TABLE 4H).

TABLE 4HHPLC results of napadisylate Type A (824511-30-B7)#RRTArea (%)#RRTArea (%)10.880.0951.0099.2120.930.1061.070.2230.960.1971.100.0540.990.15------

Napadisylate Type B

Napadisylate Type B (824511-36-A7) was obtained by slurry ~20.0 mg freebase and 16.1 mg naphthalene-1,5-disulfonic acid (charge molar ratio 1:1) in 0.5 mL IPA at RT for about 3 days and then transfer to slurry at 50° C. for about 4 days. Resulting solids were isolated by centrifugation and vacuum drying at RT overnight. The XRPD pattern was displayed inFIG.3Qand TABLE 41. As TGA/DSC curves inFIG.3Rshowed, a weight loss of 6.0% up to 80° C. and two endothermic signals at 81.7° C. and 206.0° C. (peak) were observed.1H NMR spectrum inFIG.3Sshowed that the peaks of naphthalene-I,5-disulfonic acid and IPA were observed. The molar ratio of acid/API was 0.6:1, the molar ratio of IPA/API was 0.78:1 (theoretical 8.17 wt%). HPLC purity was 99.66 area% (FIG.3Tand TABLE 4J).

TABLE 4JHPLC results of napadisylate Type B (824511-36-A7)#RRTArea (%)10.970.0721.0099.6631.070.27

Napadisylate Type C

Napadisylate Type C (824511-36-B7) was obtained by slurry ~20.0 mg freebase and 16.1 mg naphthalene-1,5-disulfonic acid (charge molar ratio of acid to freebase was 1:1) in 0.5 mL 1,4-dioxane for about 3 days and then transfer to slurry at 50° C. for about 4 days. Resulting solids were isolated by centrifugation and vacuum drying at RT overnight. The XRPD overlay was displayed inFIG.3U, with XRPD peak list of (824511-44-B2) shown in TABLE 4K. As TGA/DSC curves inFIG.3Vshowed, a weight loss of 6.7% up to 100° C. and three endothermic signals at 71.2° C., 117.0° C. and 191.0° C. (peak) were observed.1H NMR spectrum inFIG.3Wshowed that the peak of naphthalene-I,5-disulfonic acid and 1,4-dioxane were observed. The molar ratio of acid/API was 0.7:1, the molar ratio of 1,4-dioxane/API was 0.50:1 (theoretical 8.91 wt%). HPLC purity was 98.42 area% (FIG.3Xand TABLE 4L).

TABLE 4LHPLC results of napadisylate Type C (824511-44-B2)#RRTArea (%)#RRTArea (%)10.910.4240.970.3420.950.3151.0098.4230.960.2461.070.28

Napadisylate Type D

Napadisylate Type D (824511-44-B1) was discovered in the re-preparation trial of napadisylate Type C by slurry 20.2 mg freebase and 16.0 mg naphthalene-1,5-disulfonic acid (charge molar ratio of acid to freebase was 1:1) in 0.5 mL 1,4-dioxane at RT for 8 days. Resulting solids were isolated by centrifugation and vacuum drying at RT overnight. The XRPD overlay was displayed inFIG.3Ywith peak list of (824511-44-B1) shown in TABLE 4 M. As TGA/DSC curves inFIG.3Zshowed, a weight loss of 6.9% up to 100° C. and three endothermic signals at 53.9° C., 89.0° C. and 178.3° C. (peak) were observed.1H NMR spectrum inFIG.3AAshowed that the peak of naphthalene-I,5-disulfonic acid was observed. The molar ratio of acid/API was 0.7:1. No obvious solvent residual was detected. HPLC purity was 99.56 area% (FIG.3ABand TABLE 4N).

TABLE 4NHPLC results of napadisylate Type D (824511-44-B1)#RRTArea (%)#RRTArea (%)10.700.0541.0099.5620.910.0851.070.2630.970.06------

Besylate Type A

Besylate Type A (824511-30-A10) was obtained by slurry ~20.0 mg freebase and 8.9 mg benzenesulfonic acid (charge molar ratio of acid to freebase was 1:1) in 0.5 mL DCM/EtOAc (1:1, v/v) at RT for about 4 days and transferred to stir at 50° C. for about 1 day, then transferred to stir at -20° C. for about 3 days. Resulting solids were isolated by centrifugation and vacuum drying at RT overnight. Another batch of besylate Type A (824511-35-A1) was prepared using the same method and characterized. The XRPD overlay was displayed inFIG.3ACwith peak list of (824511-35-A1) shown in TABLE 40. TGA/DSC curves inFIG.3ADshowed a weight loss of 1.1% up to 90.0° C. and a weight loss of 3.3% from 90° C. to 130° C., two endothermic signals were observed with a major peak at 117.4° C. and a minor peak131.8° C. (peak).1H NMR result inFIG.3AEshowed that the peak of benzenesulfonic acid was observed. The molar ratio of acid/API was 1.0:1. No obvious solvent residual was detected. HPLC purity was 99.71 area% (FIG.3AFand TABLE 4P).

TABLE 4PHPLC results of besylate Type A (I824511-35-A1)#RRTArea (%)#RRTArea (%)10.910.0531.0099.7120.970.0541.070.18

Besylate Type B

Besylate Type B (824511-36-A10) was obtained by slurry ~20.0 mg freebase and 8.7 mg benzenesulfonic acid (charge molar ratio of acid to freebase was 1:1) in 0.5 mL IPA at RT overnight and then transferred to slurry at 50° C. for about 4 days. Resulting solids were isolated by centrifugation and vacuum drying at RT overnight. The XRPD result was displayed inFIG.3AGand TABLE 4Q. TGA/DSC curves inFIG.3AHshowed a weight loss of 0.4% up to 80.0° C. and two endothermic signals at 177.5° C. and 179.3° C. (peak).1H NMR result inFIG.3AIshowed that the peak of benzenesulfonic acid was observed. The molar ratio of acid/API was 1.0:1. No obvious solvent residual was detected. HPLC purity was 99.73 area% (FIG.3AJand TABLE 4R).

TABLE 4RHPLC results of besylate Type B (824511-36-A10)#RRTArea (%)11.0099.7321.070.27

Procedure of Freebase Isolation

Detailed procedure of freebase isolation is summarized in TABLE 5A.

TABLE 5ADetailed procedure of freebase isolationScaleProcedure9-g (824509-03-A)1. Dissolve 9.0 g of HCl salt Type A (824509-01-A) in 120 mL of water, and add 120 mL of DCM.2. Add 13.5 mL of aqueous ammonium hydroxide solution (25%~28%).3. Slurry at RT for ~17 hrs to obtain a pH of 10.35, then separate layers.4. Add additional 150 mL of water to wash the organic layer.5. Add additional 150 mL of DCM to wash the aqueous layer.6. Collect the organic layers and dry organic layer over anhydrous sodium sulfate.7. Filter and concentrate organic layer to dryness.8. Vacuum dry the solids at RT for ~1 hr.9. Collect the 18-MC freebase (7.98 g, yield*=97.5%).

Summary of Salts and Salt Forms

TABLE 5B summarizes the salts and salt forms produced and the corresponding reports with additional details.

Instruments and Methods

For XRPD analysis, a PANalytical Empyrean and X′ Pert3 X-ray powder diffract meter was used. The XRPD parameters used are listed in TABLE 5C.

TGA and DSC

TGA data was collected using a TA Q5000/Discovery TGA 5500 from TA Instruments. DSC was performed using a Discovery DSC 2500 from TA Instruments. Detailed parameters used are listed in TABLE 5D.

Agilent 1260 with DAD detector was utilized and detailed chromatographic condition is listed in TABLE 5E.

Thermo Scientific™ Dionex™ Aquion™ Ion Chromatography (IC) System 1100 with conductivity detector was utilized and detailed chromatographic condition is listed in TABLE 5F.

Solution1H NMR was collected on a Bruker 400 MHz NMR Spectrometer using DMSO-d6as the solvent.

Conclusion

An extended salt screening was performed for compound 18-MC using prepared freebase material. As the results showed, a total of 9 salt forms were found and characterized, including gentisate Type A, HBr salt Types A and B, besylate Types A and B and napadisylate Types A, B, C, and D.

Example 2 - Polymorph Screening of HCl Salt

Starting from HCl salt materials with different forms, a total of 100 polymorph screening experiments was performed using different crystallization methods including salt formation by liquid vapor diffusion/slurry at elevated temperature, evaporation at RT/high temperatures, slurry and reverse anti-solvent addition at different temperatures, slow/crash cooling, polymer induced crystallization and grinding. All the resulting solids were isolated for XRPD test, and new forms were further characterized by TGA, DSC,1H NMR and HPLC/ IC. As the results showed, 11 new forms (HCl salt Types L to V) were discovered. Further identification results showed that HCl salt Types M, P, Q, R, S, T, and U were solvates (which converted to HCl salt Type A after desolvation), HCl salt Types L, N, O,, and V were metastable forms (which converted to HCl salt Type A after air drying or room temperature (RT) storage). Characterization results of different HCl salt forms were summarized in TABLE 6A to 6C. Inter-conversion relationship among different forms were displayed inFIG.4.

To summarize, an additional polymorph screening was performed for 18-MC HCl salt. As the results showed, 11 new forms were obtained. Among the forms obtained, HCl salt Type A was thermodynamically more stable than other anhydrates at RT.

TABLE 6CCharacterization summary of HCl salt forms- metastable formsSolid form (ID)CrystallinityTGA loss, %Endothermic peak, °C.Form after treatmentIdentified resultHCl salt Type D (819246-46-A13)Low----HCl salt Type A**MetastableHCl salt Type E (819246-43-A5)High----HCl salt Type A**MetastableHCl salt Type G (819246-48-A2)High5.2 (up to 120° C.)108.7, 215.9HCl salt Type A#MetastableHCl salt Type J (819246-49-A6)High7.6 (up to 140° C.)87.6, 195.6HCl salt Type F#MetastableHCl salt Type V (824509-25-A4)High----HCl salt Type A**MetastableHCl salt Type L* (824509-05-A1)High----HCl salt Type A#MetastableHCl salt Type N* (824509-10-A1)High----HCl salt Type A#MetastableHCl salt Type O* (824509-10-A2)High----HCl salt Type A#Metastable--: No data collected due to form change after storage/drying.*: Only the new form mixed with HCl salt Type A was obtained in the screening and re-preparation.**: Air drying.#: RT storage.

Polymorph Screening

Using freebase Type A (824509-03-A), HCl salt Type A (824509-01-A and 824509-11-B), the low crystallinity HCl salt (824509-12-E) and amorphous (824509-20-A) as the starting material, a total of 100 polymorph screening experiments were conducted via various crystallization methods. Results of polymorph screening is summarized in TABLE 7A, TABLE 7B and TABLE 7C. XRPD results showed that a total of 11 forms (HCl salt Types L to V) were obtained in polymorph screening and characterization. The screening details are further detailed below.

HCl Salt Type M

HCl salt Type M (824509-05-A4) was obtained by salt formation through liquid vapor diffusion in 1,4-dioxane/MTBE. The detailed procedure was as follows: dissolve 20.0 mg freebase in 1.0 mL 1,4-dioxane at RT in a 4-mL vial. Dilute 1 mL HCŀEtOAc solution (conc. of HCl was 2 mol/L) by 3 mL MTBE in a 20-mL vial. Put the uncapped 4-mL vial into the 20-mL vial and keep the capped 20-mL vial at RT for about 4 days. Solids were isolated by air dried for characterization. The XRPD result is displayed inFIG.5Aand TABLE 7D. TGA/DSC results inFIG.5Bshowed a weight loss of 2.1% up to 100° C. and a weight loss of 12.6% from 100° C. to 180° C., DSC result showed one endothermic signal at 170.3° C. (peak).1H NMR result inFIG.5Cshowed that the peak of 1,4-dioxane was observed. The molar ratio of 1,4-dioxane/API was 0.5:1 (theoretical weight=8.92 wt%). HPLC/IC results showed that the molar ratio of acid/freebase was determined as 1.0:1 and HPLC purity was 99.45 area% (FIG.5Dand TABLE 7E).

InFIG.5E, XRPD overlay showed that after storing HCl salt Type M at RT for ~3 days, the extra peaks of HCl salt Type A (marked in red frame) were observed. VT-XRPD results inFIG.5Fshowed that after N2-drying for 20 min at 30° C., no form change was observed for HCl salt Type M. After heating sample to 100° C. under N2protection, peaks of HCl salt Type A were observed. After heating to 180° C. under N2protection, most diffraction peaks were consistent with HCl salt Type A with extra peak similar to HCl salt Type K. After cooling back to 30° C. under N2protection, most diffraction peaks were consistent with HCl salt Type A. Considering the obvious solvent amount in the sample, HCl salt Type M was speculated as a 1,4-dioxane solvate that converted to Type A upon desolvation.

TABLE 7EHPLC results of HCl salt Type M (824509-05-A4)#RRTArea (%)#RRTArea (%)10.910.0541.070.3520.970.1051.200.0531.0099.45------

HCl Salt Type P

HCl salt Type P (824509-10-A3) was obtained by adding 1.0 mL 2,2,2-trifluoroethanol solution (conc. of HCl salt was ~20 mg/mL) in 9.0 mL MIBK directly at RT and stir at RT for 4 days. Since no solids precipitated after RT stirring, the clear solution was transferred to stir at 5° C. overnight, -20° C. overnight and evaporation at RT for about 3 weeks. The resulting solids were centrifuged and air dried for characterization. The XRPD result is displayed inFIG.6Aand TABLE 8A. TGA/DSC results inFIG.6Bshowed a weight loss of 1.8% up to 80° C. and a weight loss of 21.9% from 80° C. to 150° C., DSC results showed one exothermic signal at 122.7° C. (peak) and one endothermic signal at 202.3° C. (peak).1H NMR spectrum inFIG.6Cshowed that the peak of 2,2,2-trifluoroethanol was observed. The molar ratio of 2,2,2-trifluoroethanol/API was 1.1:1 (theoretical weight=21.1 wt%). HPLC/IC results showed that the molar ratio of acid/freebase was determined as 0.9:1 and HPLC purity was 98.71 area% (FIG.6Dand TABLE 8B).

The VT-XRPD result inFIG.6Eshowed that after N2-drying for 20 min at 30° C., no form change was observed for HCl salt Type P. After heating to 100° C. and 150° C. under N2protection, form change to HCl salt Type A was observed. After heating to 210° C. under N2protection, amorphous sample was observed. XRPD overlay inFIG.6Fshowed that after heating HCl salt Type P to 80° C. and cooling back to RT, no obvious form change was observed. After 150° C. heating, HCl salt Type P converted to HCl salt Type A.1H NMR result inFIG.6Gshowed that the peak of 2,2,2-trifluoroethanol was still observed after 80° C. heating, and molar ratio of 2,2,2-trifluoroethanol/API was 1.0:1 (theoretical weight=19.2 wt%). Combined with the results of heating experiments and VT-XRPD, HCl salt Type P was speculated to be a 2,2,2-trifluoroethanol solvate that converted to Type A upon desolvation.

TABLE 8BHPLC results of HCl salt Type P (824509-10-A3)#RRTArea (%)#RRTArea (%)10.730.1081.070.2620.750.2091.170.1530.910.10101.220.0640.920.05111.240.1250.970.06121.270.0761.0098.71131.340.0971.060.05------

HCl Salt Type Q and R

HCl salt Type Q (824509-16-A4) was obtained by slurrying 20.3 mg HCl salt Type A (824509-12-E) in 0.5 mL isopentanol at 60° C. for about 4 days. Resulting solids were isolated by centrifugation and air drying. HCl salt Type R (824509-16-A3) was obtained from 2-BuOH via the same method. XRPD results were shown inFIGS.7A to7C, TABLE 9A and TABLE 9B. The two forms showed similar XRPD patterns with differences marked inFIG.7Cand peak list of (824509-16-A4) was shown in TABLE 9A.

For HCl salt Type Q (824509-16-A4), TGA/DSC results inFIG.7Dshowed a weight loss of 15.3% up to 130° C. and two endothermic signals at 82.9° C. and 141.9° C. (peak).1H NMR result inFIG.7Eshowed that the peak of isopentanol was observed. The molar ratio of isopentanol/API was 0.7:1 (theoretical weight=11.6 wt%). HPLC/IC results showed that the molar ratio of acid/freebase was determined as 1.0:1 and HPLC purity was 99.74 area% (FIG.7Fand TABLE 9C). VT-XRPD results inFIG.7Gshowed that after N2-drying for 20 min at 30° C., no form change was observed for HCl salt Type Q. After heating HCl salt Type Q to 100° C., 130° C., 160° C. and cooling back to 30° C. under N2protection, form change to a mixture of HCl salt Type A and freebase Type A (FIG.7H, peak of freebase Type A was marked in red frame) was observed. InFIG.7I,1H NMR result showed that after VT-XRPD test, no signal of isopentanol was observed. Combined with the VT-XRPD results, HCl salt Type Q was speculated as an isopentanol solvate.

For HCl salt Type R (824509-16-A3), TGA/DSC results inFIG.7Jshowed a weight loss of 15.0% up to 150° C. and one endothermic signal at 141.8° C. (peak).1H NMR result inFIG.7Kshowed that the peak of 2-BuOH was observed, the molar ratio of 2-BuOH/API was 0.8:1 (theoretical weight=13.4 wt%). HPLC/IC results showed that the molar ratio of acid/freebase was determined as 1.0:1 and HPLC purity was 99.64 area% (FIG.7Land TABLE 9D). XRPD results inFIG.7Mshowed that after heating to 170° C. and cooling back to ambient condition, HCl salt Type R converted to HCl salt Type A. InFIG.7N,1H NMR result showed that after 170° C. heating, no peak of 2-BuOH was observed. Combined with the results of heating experiment, HCl salt Type R was speculated as a 2-BuOH solvate that converted to Type A upon desolvation.

TABLE 9CHPLC results of HCl salt Type Q (824509-16-A4)#RRTArea (%)10.920.0721.0099.7431.070.20

TABLE 9DHPLC results of HCl salt Type R (824509-16-A3)#RRTArea (%)10.920.0821.0099.6431.070.2141.160.07

HCl Salt Type S

HCl salt Type S (824509-25-A2) was obtained by slurrying 19.7 mg amorphous HCl salt in 0.5 mL cyclohexanone at -20° C. for about 4 days. The resulting solids were isolated by centrifugation and the wet cake was tested by XRPD. The Type S sample turned to be gel like after air drying at RT It was re-prepared (824509-29-A3) by slurrying amorphous HCl salt in cyclohexanone at -20° C. for 7 days, and vacuum drying at RT for ~4 hours. The XRPD results are displayed inFIG.8Aand TABLE 10A. TGA/DSC results inFIG.8Bshowed a weight loss of 2.8% up to 150° C., DSC result showed two endothermic signals at 98.0° C. and 208.0° C. (peak).1H NMR results inFIG.8Cshowed that the peak of cyclohexanone was observed. The molar ratio of cyclohexanone/API was 0.15:1 (theoretical weight=3.51 wt%). HPLC/IC results showed that the molar ratio of acid/freebase was determined as 1.0:1 and HPLC purity was 99.20 area% (FIG.8Dand TABLE 10B).

XRPD results inFIG.8Eshowed that after heating Type S sample (824509-39-A2) to 150° C., most diffraction peaks were consistent with HCl salt Type A. Using DMSO-d6as solvent,1H NMR result inFIG.8Fshowed that after 150° C. heating, no peak of cyclohexanone was observed. Combined with the results of heating experiment and no obvious form transition signal in DSC, it was only obtained in a cyclohexanone system, HCl salt Type S was possibly a cyclohexanone solvate. Since the molar ratio of solvent to API was a bit low, it could be a channel solvate which may have a nonstoichiometric amount of solvent.

TABLE 10BHPLC results of HCl salt Type S (824509-29-A3)#RRTArea (%)#RRTArea (%)10.690.1150.950.1320.880.2061.0099.2030.910.0971.070.1840.930.09------

HCl Salt Type T

HCl salt Type T (824509-39-A1) was obtained by slurrying 40 mg HCl salt amorphous in 0.5 mL propionic acid at -20° C. for 4 days and drying at RT with silica gel. The XRPD results are displayed inFIG.9A,FIG.9Band TABLE 11A. TGA/DSC results inFIG.9Cshowed a weight loss of 11.1% up to 150° C., DSC results showed five exothermic signals at 91.6° C., 112.9° C., 135.5° C., 190.8° C. and 207.4° C. (peak).1H NMR spectrum inFIG.9Dshowed that the peak of propionic acid was observed. The molar ratio of propionic acid/API was 0.63:1 (theoretical weight=10.29 wt%). HPLC/IC results showed that the molar ratio of acid/freebase was determined as 1.0:1 and HPLC purity was 99.38 area% (FIG.9Eand TABLE 11B).

XRPD results inFIG.9Fshowed that after heating HCl salt Type T sample to 150° C. and cooling back to RT, most diffraction peaks were consistent with HCl salt Type A.1H NMR result inFIG.9Gshowed that amount of propionic acid decreased significantly (molar ratio of propionic acid/API was 0.06:1, theoretical weight=1.09 wt%). Combined with the results of heating experiment, HCl salt Type T was speculated as a propionic acid solvate that desolvated to Type A.

TABLE 11BHPLC results of HCl salt Type T (824509-39-A1)#RRTArea (%)#RRTArea (%)10.890.1450.990.0620.910.0961.0099.3830.960.1571.070.1440.970.05------

HCl Salt Type U

HCl salt Type U (824509-29-B4) was obtained by slurry 40 mg HCl salt amorphous in 0.5 mL benzylalcohol/methyl acetate (1:1, v/v) at -20° C. for 3 days, and isolated by centrifugation and vacuum dried at 50° C. ~3 hrs, and this sample was used for characterization. The XRPD results were displayed inFIG.10A, and TABLE 12A. TGA/DSC results inFIG.10Bshowed a weight loss of 2.5% up to 70° C. and a weight loss of 16.4% from 70° C. up to 120° C., DSC result showed one endothermic signal at 132.9° C. (peak).1H NMR spectrum inFIG.10Cshowed that the peak of benzyl alcohol was observed. The molar ratio of benzyl alcohol/API was 1.0:1 (theoretical weight=21.08 wt%). HPLC/IC results showed that the molar ratio of acid/freebase was determined as 0.9:1 and HPLC purity was 99.67 area% (FIG.10Dand TABLE 12B).

XRPD results inFIG.10Eshowed that after heating HCl salt Type U to 120° C., an extra peak representative of HCl salt Type A was observed, and after heating to 150° C., most diffraction peaks were consistent with HCl salt Type A.1H NMR result inFIG.10Fshowed that after heating to 150° C., the peak of benzyl alcohol in the sample was decreased significantly (molar ratio of benzyl alcohol/API was 0.03:1, theoretical weight=0.79 wt%). Combined with the results of heating experiment, HCl salt Type U was speculated to be a benzyl alcohol solvate that converted to Type A upon desolvation.

TABLE 12BHPLC results of HCl salt Type U (824509-29-B4)#RRTArea (%)10.730.0620.910.0730.970.0641.0099.6751.070.15

Metastable Forms

HCl Salt Type V

HCl salt Type V (824509-25-A4) was obtained by slurry 20.0 mg HCl salt amorphous in 0.5 mL trifluoroethanol/m-xylene (1:1, v/v) at -20° C. overnight, then the clear solution was transferred to anti-solvent addition (EtOAc, 4.5 mL) at RT and stir at 5 and -20° C. to produce more solids for 5 days. Resulting solids were isolated by centrifugation and air dried in desiccator at RT with silica gel for ~3 hrs. The XRPD overlay of the wet sample was displayed inFIG.11Aand peak list of (824509-25-A4) was shown in TABLE 13. XRPD result inFIG.11Bshowed that after air drying the wet cake for 3 hrs, the sample converted to HCl salt Type A completely. Since HCl salt Type V was not obtained in the re-preparation trials (details refer to Section 4.6), no more characterization data was collected.

HCl Salt Type L/N/O Mixtures

Sample (824509-05-A1,FIG.12Aand TABLE 14A) was obtained by salt formation through liquid vapor diffusion in MIBK/n-pentane system. The detailed procedure was as follows: dissolve 19.9 mg freebase in 1.0 mL MIBK at RT in a 4-mL vial. Dilute 1 mL HCŀEtOAc solution (conc. of HCl was 2 mol/L) by 3 mL n-pentane in a 20-mL vial. Put the uncapped 4-mL vial into the 20-mL vial and keep the capped 20-mL vial at RT for about 4 days. Solids were isolated by centrifugation and air dried for characterization.

XRPD pattern inFIG.12Bshowed that the sample (824509-05-A1) had strong extra peaks compared with HCl salt Type A, which was assigned as HCl salt Type A+L. After storing the sample (824509-05-A1) at RT for ~22 days, it converted to HCl salt Type A completely. Since no pure Type L was obtained in the screening, no more characterization was performed.

HCl salt Type A+N (824509-10-A1,FIG.12Cand TABLE 14B) was obtained by adding 1.0 mL 2,2,2-trifluoroethanol solution (conc. of HCl salt was ~20 mg/mL) in 9.0 mL toluene directly at RT and stir at RT for 1 days. Since no solids precipitated after RT stirring, the clear solution was transferred to stir at 5° C. overnight, -20° C. overnight and evaporation at RT for about 5 days to dryness. XRPD pattern inFIG.12Dshowed that the sample (824509-10-A1) had some strong extra peaks compared with HCl salt Type A, which was assigned as HCl salt Type A+N. After storage the sample (824509-10-A1) at RT for ~21 days, it converted to HCl salt Type A completely. Since pure Type N was not obtained, additional characterization was not performed.

HCl salt Type A+O (824509-10-A2,FIG.12Eand TABLE 14C) was obtained by reverse anti-solvent addition in 2,2,2-trifluoroethanol/THF systems. XRPD result showed that the sample (824509-10-A2) had some strong extra peaks compared with HCl salt Type A, assigned as HCl salt Type A+O. After storing the sample (824509-10-A2) at RT for ~19 days, it converted to HCl salt Type A completely (FIG.12F). Since pure Type O was not obtained, additional characterization was not performed.

Characterization Information of HCl Salt Type A to K

A summary of information about HCl salt Type A to K is shown in TABLE 15.

Conclusion

An additional polymorph screening was performed for 18-MC HCl salt. As the characterization results showed, 11 new forms were obtained. Among the forms obtained, HCl salt Type A was still the most thermodynamically stable form at RT based on desolvation and physical form conversion data.

18-MC sulfate Type A was first obtained as described in Example 1. Polymorph screening of 18-MC sulfate was performed to better understand polymorphism of the salt.

Sulfate material was first prepared using 18-MC freebase and used as starting material for polymorph screening. In the screening, different crystallization methods including temperature cycling and slurry conversion at different temperatures were used, and a total of 30 experiments were conducted. Solids from screening were isolated for XRPD. New forms were further characterized by TGA, DSC and HPLC/IC. As the characterization and identification results showed, five new forms (sulfate Type B-F) were discovered. Identification results indicated that sulfate Type A and Type F were anhydrates, Type D was a hydrate, Type E was a hydrate or anhydrate, Type B was an ACN solvate (converted to sulfate Type E after storage) and Type C was a metastable form (converted to sulfate Type A after air drying). Characterization results are summarized in TABLE 15 and XRPD patterns of different forms are displayed inFIG.13.

To summarize, a brief polymorph screen was performed and a total of six forms of the sulfate were discovered.

Preparation of Sulfate Starting Material

The preparation procedure of sulfate Type A (824511-04-A) was as follows: 2 g of freebase Type A (824509-01-A was weighed, 20 mL of EtOAc was added to prepare a suspension. 1.36 mL of 4 M H2SO4was diluted by 20 mL of EtOAc. The diluted H2SO4solution was added into the suspension under magnetic stirring dropwise. The sample was transferred to temperature cycling (50° C.~5° C., 3 cycles, one cycle: heat to 50° C. at 4.5° C./min, isothermal at 50° C. for 30 min; cool to 5° C. at 0.1° C./min, isothermal at 5° C. for 30 min; keep slurry at 5° C. at last). After XRPD confirmation, the solids were isolated by vacuum filtration and vacuum dried at RT for one day. As a result, about 1.77 g of sulfate Type A (824511-04-A) was obtained.

The XRPD pattern of prepared sulfate Type A inFIG.14Awas consistent with sulfate Type A reference (819246-23-A2). TGA/DSC curves inFIG.14Bshowed a weight loss of 1.6% up to 120.0° C. and overlapped endothermic peaks around 176.3° C. (peak).1H NMR (DMSO-d6as solvent) result inFIG.14Cshowed that EtOAc was not observed. The molar ratio of acid/freebase was determined as 0.97:1 by HPLC/IC and HPLC purity was tested to be 99.77 area% (FIG.14Dand TABLE 16A).

VT-XRPD results inFIG.14Eshowed that after N2-drying sulfate Type A (824511-04-A) for 20 min at 30° C. or heating sample to 120° C. and cooling back to 30° C. under N2protection, no obvious form change was observed, indicating sulfate Type A was an anhydrate.

Approximate solubility values of sulfate Type A (824511-04-A) were estimated in 10 solvents to guide the solvent selection in the polymorph screening, with results summarized in TABLE 16B.

TABLE 16AHPLC results of sulfate Type A (824511-04-A)#RRTArea (%)10.970.0521.0099.7731.070.18

TABLE 16BApproximate solubility of sulfate Type A (824511-04-A) at RTSolventSolubility (mg/mL)SolventSolubility (mg/mL)MeOHS>44.0DCMS<2.1EtOH20.0<S<40.0EtOAcS<2.0ACN20.0<S<40.0THFS<1.9Acetone1.8<S<6.0CHCl3S<1.9TolueneS<2.2n-HeptaneS<1.8Procedure: weigh ~2 mg solids into each 3-mL glass vial, add in corresponding solvent stepwise and sonicate or oscillate to see if solids dissolved completely. Stop adding solvent till the solids dissolves or total volume reaches 1.0 mL. Calculate the approximate solubility based on solvent volume.

XPRD data for sulfate Type A is in TABLE 16C.

Polymorph Screening

Using prepared sulfate Type A (824511-04-A, anhydrate) as the starting material, a total of 30 polymorph screening experiments were conducted via various crystallization methods. Results of polymorph screening is summarized in TABLE 17A. Results showed that a total of 6 forms (sulfate Type A~F) were obtained in the polymorph screening, including 2 anhydrates (sulfate Type A/F), 1 hydrate (sulfate Type D), 1 solvate (sulfate Type B), and 1 metastable form (sulfate Type C). Sulfate Type E was a hydrate or anhydrate, while no further identification was conducted due to the failure of re-preparation trials. Characterization data of obtained forms was summarized in TABLE 15 and the XRPD overlays of these forms were displayed inFIG.13.

TABLE 17ASummary of polymorph screening experiments of sulfateMethodNo. of ExperimentResultsTemperature Cycling5Sulfate Type A/B/CSlurry at RT20Sulfate Type A/B/D, amorphous, gelSlurry at 50° C.5Sulfate Type A/BTotal30Sulfate Type A/B/C/D, amorphous, gelNote: Sulfate Type E and F were found during form identification.

Anhydrate and Hydrate

Sulfate Type D and F

Sulfate Type D (824511-12-A17) was obtained by slurrying 20.3 mg sulfate Type A (824511-04-A) in THF/H2O (981:19, v/v, aw≈0.2) at RT for about one week and isolating solids by centrifugation and air drying. The XRPD result is displayed inFIG.15Aand Table 17B. TGA/DSC results inFIG.15Bshowed a weight loss of 2.3% up to 100.0° C. and two endothermic peaks at 76.1° C. and 194.9° C. (peak). Using MeOH-d4 as solvent,1H NMR result inFIG.15Cshowed that no peak of THF was observed. HPLC purity of the sample was determined as 99.79 area% (FIG.15Dand TABLE 17C). InFIG.15E, XRPD result showed that after drying sulfate Type D by N2for 20 min at 30° C., a new form was observed and assigned as sulfate Type F (FIG.15Fand TABLE 17D). After heating to 100° C. and cooling back to 30° C. under N2protection, the sample remained sulfate Type F. After exposure to ambient condition for ~30 min, sulfate Type F converted back to sulfate Type D. Thus, sulfate Type D was speculated to be a hydrate (molar ratio of water to salt was 0.6, calculation based on TGA loss) and sulfate Type F was an anhydrate. Since sulfate Type F was not physically stable at ambient conditions, no more characterization was conducted.

TABLE 17CHPLC results of sulfate Type D (824511-12-A17)#RRTArea (%)10.950.0821.0099.7931.070.13

Sulfate Type E

Sulfate Type E (824511-11-A3-0315) was obtained by storing sulfate Type B (824511-11-A3) at RT in a sealed vial for ~12 days. The XRPD result was displayed inFIG.15Gand TABLE 17E. TGA/DSC curves inFIG.15Hshowed a weight loss of 3.0% up to 120.0° C., three endothermic signals at 74.1° C., 116.8° C. and 147.0° C. (peak). XRPD overlay inFIG.15lshowed that after heating to 100° C. and cooling back to ambient conditions, no form change was observed. Using DMSO-d6as solvent,1H NMR result of sulfate Type E from heating experiment inFIG.15Jshowed that the peak of ACN was observed. The molar ratio of ACN/API was 0.03:1 (theoretical weight=0.3 wt%). Combined with the results of heating experiment, sulfate Type E was speculated to be a hydrate or anhydrate. Since sulfate Type E was not obtained in the re-preparation trials, no more characterization data were collected.

Sulfate Type B (824511-11-A3) was obtained by slurrying sulfate Type A (824511-04-A) in ACN with temperature cycling (suspend 20.3 mg sulfate in 0.5 mL ACN at RT and then transfer to temperature cycling (3 cycle): ramp to 50° C. at a rate of 1° C./min, isothermal for 120 min, cool to 5° C. at a rate of 0.1° C./min, isothermal for 120 min). Resulting solids were isolated by centrifugation and air drying. The XRPD result is displayed inFIG.15Kand TABLE 17F. TGA/DSC results inFIG.15Lshowed a weight of 6.6% up to 100.0° C., four endothermic signals at 65.4° C., 115.9° C., 146.8° C. and 188.4° C. (peak). Using DMSO-d6as solvent,1H NMR result inFIG.15Mshowed that the peak of ACN was observed. The molar ratio of ACN/API was 0.6:1 (theoretical weight=5.4 wt%, similar to weight loss in TGA before 100.0° C.). HPLC purity of the sample was determined as 99.64 area% (FIG.15Nand TABLE 17G). The molar ratio of acid/freebase was determined as 0.9:1 by HPLC/IC. XRPD overlay inFIG.15Oshowed that after storage sulfate Type B at RT for ~12 days, form change to sulfate Type E was observed and weight loss before 100.0° C. decreased from 6.6% to 2.6% (FIG.15P). Considering that sulfate Type B was only obtained in ACN conditions in the screening experiments, sulfate Type B was possibly an ACN solvate.

TABLE 17GHPLC results of sulfate Type B (824511-11-A3)#RRTArea (%)#RRTArea (%)10.890.0941.0099.6420.930.0751.070.1130.950.08------

Metastable Form

Sulfate Type C (824511-11-A4) was obtained by slurrying sulfate Type A (824511-04-A) in 1,4-dioxane with temperature cycling (suspend 20.3 mg sulfate in 0.5 mL 1,4-dioxane at RT and then transfer to temperature cycling (3 cycle): ramp to 50° C. at a rate of 1° C./min, isothermal for 120 min, cool to 5° C. at a rate of 0.1° C./min, isothermal for 120 min). Resulting solids were isolated by centrifugation and air drying. The XRPD result is displayed inFIG.15Qand TABLE 17H. The XRPD overlay inFIG.15Rshowed that after air drying at RT for ~30 min, sulfate Type C converted to sulfate Type A, which indicated that sulfate Type C was a metastable form. Since sulfate Type C was not obtained in the re-preparation trials, no more characterization data were collected.

Conclusion

A brief polymorph screening of 18-MC sulfate was performed and a total of six salt forms were obtained.

18-MC oxalate Type A was first obtained as described in Example 1. Polymorph screening of 18-MC oxalate was performed to better understand polymorphism of the salt.

Oxalate material was prepared using 18-MC freebase and used as starting material for polymorph screening. X-ray powder diffraction (XRPD) result showed that the prepared oxalate displayed a different XRPD pattern from oxalate Type A, which was assigned as oxalate Type B, and further characterized by TGA, DSC and HPLCIC. Based on the limited weight loss in TGA and neat DSC before decomposition, both oxalate Type A and oxalate Type B were speculated as anhydrates.

Using prepared oxalate Type B as the starting material, a total of 30 polymorph screening experiments were conducted with different crystallization methods including temperature cycling and slurry conversion at different temperatures. Solids from the screening experiments were isolated for XRPD testing. As the results showed, only oxalate Type B was obtained. Characterization results of oxalate Type A and B were summarized in TABLE 18 and XRPD patterns of the two forms were displayed inFIG.16A.

To summarize, a brief polymorph screening experiments were performed and only oxalate Type B was observed.

Preparation of Oxalate Starting Material

Preparation procedure of oxalate was as follows: weigh 2.0 g of freebase Type A (824509-03-A) and 0.7 g of oxalic acid dihydrate into 500 mL of EtOAc and transfer the suspension to slurry at RT for ~5 days. Resulting solids were isolated by vacuum filtration and vacuum drying at RT overnight. About 2.4 g of oxalate sample (824511-04-C) was obtained. XRPD comparison results inFIG.16Ashowed that the obtained oxalate displayed a different pattern than oxalate Type A (819246-23-A20), and it was named as oxalate Type B, with XRPD peak list shown in TABLE 19A. TGA/DSC curves inFIG.16Bshowed a weight loss of 1.1% up to 150.0° C. and one endothermic peak at 167.1° C. (onset). Proton nuclear magnetic resonance (1H NMR) result inFIG.16Cshowed that peak of EtOAc was observed. The molar ratio of EtOAc/API was 0.08:1 (theoretical weight=1.5 wt%). HPLC/IC results showed that the molar ratio of acid/freebase was determined as 1.08:1 and HPLC purity was 98.95 area% (FIG.16Dand TABLE 19B). Considering the limited weight loss and neat DSC curve before decomposition, oxalate Type B was speculated to be an anhydrate.

Approximate solubility of oxalate Type B (824511-04-C) was estimated in 10 solvents to guide the solvent selection in polymorph screening of oxalate, and the results were summarized in TABLE 19C.

TABLE 19BHPLC results of oxalate Type B (824511-04-C)#RRTArea (%)#RRTArea (%)10.290.3250.930.0620.830.0560.970.0830.890.1071.0098.9540.930.1681.070.28

TABLE 19CApproximate solubility of oxalate Type B (824511-04-C) at RTSolventSolubility (mg/mL)SolventSolubility (mg/mL)MeOHS>44.0THF6.3<S<19.0H2OS>38.0CHCl32.3<S<7.7ACN7.7<S<23.0DCM2.1<S<7.0EtOH7.0<S<21.0n-HeptaneS<2.1Acetone7.0<S<21.0EtOAcS<1.8

Procedure: weigh ~2 mg solids into each 3-mL glass vial, add in corresponding solvent stepwise and sonicate or oscillate to see if solids dissolved completely. Stop adding solvent till the solids dissolves or total volume reaches 1.0 mL. Calculate the approximate solubility based on solvent volume.

Polymorph Screening

Using oxalate Type B (824511-04-C) as the starting material, a total of 30 experiments were conducted using different crystallization methods. XRPD results showed that oxalate Type B was the only form obtained from the screen. The results of polymorph screening are summarized in TABLE 20A, and only oxalate Type B was observed.

TABLE 20ASummary of polymorph screening experiments of oxalateMethodNo. of ExperimentResultsTemperature Cycling5Oxalate Type BSlurry at RT20Oxalate Type BSlurry at 50° C.5Oxalate Type BTotal30Oxalate Type B

Temperature cycling experiments were conducted in 5 solvent systems. About 20 mg of starting material (824511-04-C) was suspended in an HPLC vial with 0.5 mL of corresponding solvents listed in TABLE 20B. Cycling procedure: ramped to 50° C. at a rate of 4.5° C./min, kept the temperature at 50° C. for 30 min; cooled down to 5° C. at a rate of 0.1° C./min and kept the temperature at 5° C. for 30 min. After three cycles, the obtained solids were isolated by centrifugation and air dried for XRPD analysis. Results were summarized in TABLE 20B, and only oxalate Type B was observed.

TABLE 20BSummary of temperature cycling experiments of oxalateExperiment IDSolventSolid form824511-14-A1EtOHOxalate Type B824511-14-A2AcetoneOxalate Type B824511-14-A3ACNOxalate Type B824511-14-A42-MeTHFOxalate Type B824511-14-A5CHCl3Oxalate Type B

Slurry Conversion at RT

Slurry conversion experiments were conducted at RT in 20 different solvent systems. About 20 mg of starting material (824511-04-C) was suspended in an HPLC vial with 0.5 mL of corresponding solvents listed in TABLE 20C. If the solid dissolved then more solids were added until a suspension was obtained. The suspensions were stirred at RT using magnetic stirring with a speed of ~750 rpm. After about 5 days, the remaining solids were isolated by centrifugation and air dried for XRPD analysis. If a clear solution was obtained, the sample was cooled to 5° C. If it was still clear, the solution was cooled to -20° C. If there was still no precipitate, the solution was allowed to evaporate in an open vial at RT to obtain solids. Results are summarized in TABLE 20C, and only oxalate Type B was observed.

Slurry conversion experiments were conducted at 50° C. in 5 solvent systems. About 20 mg of starting material (824511-04-C) was suspended in an HPLC glass vial with 0.5 mL of corresponding solvents listed in TABLE 20D. Samples were stirred at 50° C. for 5 days. If there was no solid after the slurry, the sample was cooled to 5° C. Resulting solids were isolated by centrifugation and air dried for XRPD analysis. Results are summarized in TABLE 20D, and only oxalate Type B was observed.

TABLE 20DSummary of slurry conversion experiments at 50° C. of oxalateExperiment IDSolventSolid form824511-16-A1IPAOxalate Type B824511-16-A2AcetoneOxalate Type B824511-16-A3TolueneOxalate Type B824511-16-A4EtOAcOxalate Type B824511-16-A5ACNOxalate Type B

XPRD data for oxalate Type A is shown in TABLE 20E.

Conclusion

A brief polymorph screening of 18-MC oxalate was performed and one new form was obtained.

18-MC mesylate Type A was first obtained as described in Example 1. Polymorph screening of 18-MC mesylate was performed to better understand polymorphism of the salt.

Mesylate material was first prepared using 18-MC freebase and used as starting material for polymorph screening. In the screening, different crystallization methods including temperature cycling and slurry conversion at different temperatures were used, and a total of 30 experiments were conducted. Solids from the screening were isolated for XRPD testing. As the results showed, two new forms, assigned as mesylate Types B and C, with weak crystallinity were observed. Attempts were made to reprepare the two new forms for further characterization. However, they were not found to be physically stable at ambient conditions and easily converted to gels/oils. Additional data were not collected. XRPD patterns of different forms are displayed inFIG.17, and characterization results are summarized in TABLE 21.

To summarize, brief polymorph screening experiments were performed and a total of three forms of mesylate were discovered.

TABLE 21Characterization summary of mesylate salt formsSolid form (ID: 824511-)TGA loss (%)Endotherm (°C., peak)Solvent residual (wt%)#Molar ratio#(acid/FB)HPLC purity (area%)Speculated formMesylate Type A (23-B)1.7 up to 80° C.89.2, 171.50.5 (EtOAc)1.098.86AnhydrateMesylate Type B (32-A1)----------Metastable*Mesylate Type C (32-A2)----------Metastable*FB: Freebase.--: No data collected due to limited sample amount and failure of re-preparation trials.#: Calculation based on1H NMR result.*: The solid turn to gel-like after air drying at RT in a few minutes.

Preparation of Mesylate Starting Material

Mesylate Type A (824511-23-B) was prepared as follows: 1.0 g of freebase (824509-21-A) was weighed into a 20-mL glass vial. Approximately 5 mL of EtOAc was added to prepare a suspension. Methanesulfonic acid (264.6 mg) was mixed with 5 mL of EtOAc, and the acid solution was added to the freebase suspension dropwise while stirring with a magnetic stirrer. The sample became gel like, which was then stirred at RT for 3 days. The sample was then used for a temperature cycling experiment to improve crystallinity (50° C.~5° C., 2 cycles, one cycle: heat to 50° C. at 4.5° C./min, isothermal at 50° C. for 30 min; cool to 5° C. at 0.1° C./min, isothermal at 5° C. for 30 min). After XRPD confirmation on a slurry sample, the sample was centrifuged and vacuum dried at RT overnight. As a result, 1.11 g of mesylate Type A (824511-23-B) was obtained. The XRPD pattern of prepared mesylate Type A and reference is shown inFIG.18AandFIG.18B. XRPD peak list of (824511-23-B) is shown in TABLE 22A. Thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC) curves inFIG.18Cshowed a weight loss of 1.7% up to 80.0° C. and two endothermic signals at 89.2° C. and 171.5° C. (peak). Proton nuclear magnetic resonance (1H NMR) result inFIG.18Dshowed that the peak of methanesulfonic acid and EtOAc was observed. The molar ratio of methanesulfonic acid/API was 1:1, the molar ratio of EtOAc/API was 0.027:1(theoretical weight=0.5 wt%). HPLC purity was 98.86 area% (FIG.18Eand TABLE 22B).

VT-XRPD results inFIG.18Fshowed that after N2drying mesylate Type A (824511-23-B) for 20 min at 30° C., no form change was observed. After heating mesylate Type A sample to 100° C. and cooling back to 30° C. under N2protection, no obvious form change was observed, indicating that mesylate Type A was an anhydrate.

The approximate solubility of mesylate Type A (824511-23-B) was estimated in 10 solvents to guide the solvent selection in polymorph screening of the mesylate, with results summarized in TABLE 22C.

TABLE 22BHPLC results of mesylate Type A (824511-23-B)#RRTArea (%)#RRTArea (%)10.900.1361.0098.8620.910.0571.070.2130.930.1481.090.0540.960.3691.100.0550.980.15------

TABLE 22CApproximate solubility of mesylate Type A (824511-23-B) at RTSolventSolubility (mg/mL)SolventSolubility (mg/mL)CHCl3S>42.0*EtOH20.0<S<40.0H2OS>42.0*AcetoneS<2.3MeOHS>40.0EtOAcS<2.2DCM23.0<S<46.0*n-HeptaneS<2.0ACN21.1<S<42.0*THFS<2.0Procedure: weigh ~2 mg solids into each 3-mL glass vial, add in corresponding solvent stepwise and sonicate or oscillate to see if solids dissolve completely. Stop adding solvent when the solids dissolves or total volume reaches 1.0 mL. Calculate the approximate solubility based on solvent volume.*: most of the solids were dissolved with very limited solids on the wall of the vial (not dissolved when adding more solvent). The actual solubility might be different from the test value.

Polymorph Screening

Using mesylate Type A (8245511-23-B) as the starting material, a total of 30 experiments of polymorph screening were conducted using different methods. Characterization results showed that a total of 3 forms (mesylate Types A, B, and C) were obtained, including one anhydrate (mesylate Type A) and two metastable forms (mesylate Types B and C). Since mesylate Types B and C were metastable forms and could not be re-prepared, additional data could not be collected. Results of the screening experiments are summarized in TABLE 23A. Characterization data of obtained forms was summarized in TABLE 21 and the XRPD overlays of these forms were displayed inFIG.17.

Temperature cycling experiments were conducted in 5 solvent systems. About 20 mg of starting material (824511-23-B) was suspended in an HPLC vial with 0.5 mL of corresponding solvents listed in TABLE 23B. Cycling procedure: ramped to 50° C. at a rate of 4.5° C./min, kept the temperature at 50° C. for 30 min; cooled down to 5° C. at a rate of 0.1 C/min and kept the temperature at 5° C. for 30 min. After three cycles, store the samples at 5° C. before isolation by centrifugation and air drying for XRPD analysis. Results are summarized in TABLE 23B. Results showed that mesylate Type A and amorphous were obtained.

TABLE 23BSummary of temperature cycling experiments of mesylateExp. IDSolventResults824511-26-A1EtOHAmorphous*824511-26-A2lPAcMesylate Type A824511-26-A3ACNAmorphous*824511-26-A4AcetoneAmorphous#824511-26-A5THFMesylate Type A*: Clear solution after temperature cycle→slurry at -20° C.→evaporation at RT→vacuum dried.#: Solid was obtained after -20° C. stirring and it dissolved quickly during solid isolation for XRPD test at RT.

Slurry Conversion at RT

Slurry conversion experiments were conducted at RT in 20 different solvent systems. About 20 mg of starting material (824511-23-B) was suspended in an HPLC vial with 0.5 mL of corresponding solvents listed in TABLE 23C. If the solid was dissolved then more solids were added until a suspension was obtained. The suspension was slurried at RT using magnetic stirring with the speed of ~750 rpm. After ~5 days, the remaining solids were isolated by centrifugation and air dried for XRPD analysis. If a clear solution were obtained, the sample was slurried at 5° C. If it remained clear, the sample was slurried at -20° C. If there was still no precipitate, the solution was evaporated in an open vial at RT. Results are summarized in TABLE 23C. Results showed that mesylate Type A, Type B, Type C, amorphous, gel and oil were obtained.

Slurry conversion experiments were conducted at 50° C. in 5 solvent systems. About 20 mg of starting material (824511-23-B) was suspended in an HPLC glass vial with 0.5 mL of corresponding solvents listed in TABLE 23D and stirred at 50° C. If there was no solid observed, the sample was slurried at 5° C. The resulting solids were isolated by centrifugation and air dried for XRPD analysis. Results are summarized in TABLE 23D. Results showed that mesylate Type A, amorphous, oil/gel were obtained.

TABLE 23DSummary of slurry conversion experiments at 50° C. of mesylateExperiment IDSolvent, v:vResults824511-28-A1CHCl3/n-heptane, 1:1Oil/gel**824511-28-A2ACNAmorphous*824511-28-A3IPAMesylate Type A#824511-28-A4TolueneMesylate Type A824511-28-A52-MeTHFMesylate Type A*: Clear solution from 50° C. → slurry at 5° C. → slurry at -20° C. → evaporation at RT.**: Turn to gel when slurry at 50° C. → transfer to temperature cycling.#: Solid was obtained after -20° C. stirring and it dissolved quickly during solid isolation for XRPD test at RT.

Re-Preparation Trials of Mesylate Forms

The detailed procedures and the results of re-preparation trials of mesylate are summarized in TABLE 23E.

TABLE 23ESummary of re-preparation trials of mesylate formsTarget FormExp. ID (824511-)ProcedureResultsMesylate Type A44-A1Dissolve 26.5 mg methanesulfonic acid with 0.5 mL EtOAc, add the acid solution into the vial with 99.7 mg freebase Type A (824509-03-A) dissolved in 0.5 mL EtOAc dropwise, transfer it to slurry at RT for 2 days.Mesylate Type AMesylate Type B32-A1Weigh ~50 mg of mesylate Type A (824511-23-B) in 0.5 mL of acetone and slurry at -20° C. for 5 days.Mesylate Type B*32-B1Dissolve ~8.3 mg methanesulfonic acid with 0.25 mL acetone, add the acid solution into the vial with ~30 mg freebase Type A (824509-24-A, refer to report CP827U04-01) dissolved in 0.25 mL acetone drop by drop, transfer it to slurry at -20° C., then clear solution was formed and transferred to evaporate at RT to obtain solids.Gel47-A1Mesylate Type A (824511-44-A1, 100-mg scale) was obtained by slurrying methanesulfonic acid and freebase (824509-03-A, refer to report CP827U04-01) in EtOAc (charge molar ratio 1:1). Weigh ~50 mg of the obtained mesylate Type A in 0.5 mL of acetone. Transfer the sample to slurry at -20° C. for 1 week.Mesylate Type AMesylate Type C32-A2Weigh ~50 mg of mesylate Type A (824511-23-B) in 0.5 mL of THF:H2O (981:19, v/v). Transfer the sample to slurry at -20° C. for 5 days.Mesylate Type C*32-B2Dissolve ~8.3 mg methanesulfonic acid with 0.25 mL THF:H2O (v:v,981:19), add the acid solution into the vial with ~30 mg freebase Type A (824509-24-A) dissolved in 0.25 mL acetone dropwise quickly (completed in a few seconds), transfer it to slurry at 20° C. for 2 weeks.Freebase Type A47-A2Mesylate Type A (824511-44-A1, 100-mg scale) was obtained by slurrying methanesulfonic acid and freebase (824509-03-A) in EtOAc (charge molar ratio 1:1). Weigh ~50 mg of the obtained mesylate Type A in 0.5 mL of THF:H2O (981:19, v/v). Transfer the sample to slurry at -20° C. for 1 week.Mesylate Type A+C

Conclusion

A brief polymorph screening of 18-MC mesylate was performed and a total of three salt forms were obtained.

18-MC HBr salt Type A and Type B were obtained as described in Example 1. Polymorph screening of 18-MC HBr salt was performed to have better understand polymorphism of the salt.

The HBr salt was prepared using 18-MC freebase and used as the starting material for polymorph screening. In the screening, different crystallization methods including temperature cycling and slurry conversion at different temperatures were used, and a total of 30 experiments were conducted. Solids from screening were isolated for XRPD testing. New forms were further characterized by TGA, DSC,1H NMR and HPLC/IC. As the characterization results showed, two new forms (HBr salt Types C and D) were discovered. Identification results indicated that HBr salt Type A was an anhydrate, HBr salt Type B was a hydrate or solvate, HBr salt Type C was a hydrate or anhydrate and HBr salt Type D was a solvate. HBr salt Types B, C, and D could convert to HBr salt Type A after storage or heating experiments. Characterization results are summarized in TABLE 24 and XRPD patterns of different forms are displayed inFIG.19.

To summarize, a brief polymorph screening experiments were performed and a total of four forms of HBr salt were discovered.

Preparation of HBr Salt Starting Material

Preparation procedure of HBr salt (824511-29-B) was as follows: 1.0 g of freebase (824509-24-A) was weighed into a 20-mL glass vial, and 7 mL of IPA were added to prepare a suspension. 554.8 mg of 40% HBr was added to 5 mL of IPA. The acid solution was added to the freebase suspension dropwise while stirring with a magnetic stirrer. The clear solution was slurried at RT for 3 days. The resulting sample was centrifuged and vacuum dried at RT overnight. About 1.08 g of HBr salt (824511-29-B) was obtained and it was consistent with HBr salt Type A reference (824511-01-E10), described in EXAMPLE 1. The XRPD result is shown inFIG.20A, with an overly inFIG.20B. TGA/DSC curves inFIG.20Cshowed a weight loss of 3.9% up to 170.0° C. and one endothermic peak at 203.8° C. (peak). HPLC/IC results showed that the molar ratio of acid/freebase was determined as 1.0:1 and HPLC purity was 99.43 area% (FIG.20Dand TABLE 25A). Based on the VT-XRPD results inFIG.20E, no form change of HBr salt Type A was observed when drying or heating the sample under N2to higher temperatures and cooling back to RT. Combining with neat DSC curve, HBr salt was speculated to be an anhydrate.

Approximate solubility of HBr salt Type A (824511-29-B) was estimated in 10 solvents to guide the solvent selection in polymorph screening of HBr salt, with data summarized in TABLE 25B.

TABLE 25AHPLC results of HBr salt Type A (824511-29-B)#RRTArea (%)#RRTArea (%)10.880.0840.980.0620.900.1151.0099.4330.950.2361.070.09

TABLE 25BApproximate solubility of HBr salt Type A (824511-29-B) at RTSolventSolubility (mg/mL)SolventSolubility (mg/mL)MeOHS>42.0EtOAcS<2.1EtOH7.0<S<21.0CHCl3S<2.1 *Acetone2.2<S<7.3DCMS<2.1 *H2OS<2.1n-HeptaneS<2.0ACNS<2.1 *THFS<2.0*: limited solids observed after 1.0 mL solvent addition.Procedure: weigh ~2 mg solids into each 3-mL glass vial, add in corresponding solvent stepwise and sonicate or oscillate to see if solids dissolved completely. Stop adding solvent till the solids dissolves or total volume reaches 1.0 mL. Calculate the approximate solubility based on solvent volume.

Polymorph Screening

Using HBr salt Type A (824511-29-B) as the starting material, a total of 30 polymorph screening experiments were conducted via various crystallization methods. Results of the polymorph screening are summarized in TABLE 25C. The XRPD results showed that two new forms (HBr salt Types C and D) were obtained. Characterization data of the forms are summarized in TABLE 24 and the XRPD overlay of these forms are displayed inFIG.19.

TABLE 25CSummary of polymorph screening experiments of HBr saltMethodNo. of ExperimentResultsTemperature cycling5HBr salt Type ASlurry at RT13HBr salt Type A/C/D, freebase Type A+extra peakSlurry at 50° C.12HBr salt Type ATotal30HBr salt Type A/C/D, freebase Type A+extra peak

HBr Salt Type A

HBr salt Type A was identified as an anhydrate. Characterization data on Type A reference was shown above.

HBr Salt Type B

HBr salt Type B (824511-01-D10) was obtained as described in Example 1 by slurring freebase and HBr (charge molar ratio 1:1) in 1,4-dioxane at RT. Another batch of HBr salt Type B (824511-10-A1) was prepared using the same method for characterization, and the XRPD results are displayed inFIG.20FandFIG.20G. As TGA/DSC curves inFIG.20Hshowed, a weight loss of 14.2% up to 150° C., two endothermic signals at 104.3° C., 140.3° C. (peak) and one exothermic signal at 177.4° C. (peak) were detected.1H NMR spectrum in FIGURE showed that peak of 1,4-dioxane was observed. The molar ratio of 1,4-dioxane/API was 0.6:1 (theoretical 10.8 wt%). HPLC/IC results showed that the molar ratio of acid/freebase was determined as 0.9:1 and HPLC purity was 99.27 area% (FIG.20Jand TABLE 25D). Considering that HBr salt Type B partially converted to HBr salt anhydrate Type A (peak marked) after RT storage for about 20 days (FIG.20K), and amount of solvent detected in1H NMR was similar to TGA loss, HBr salt Type B was possibly a solvate or hydrate which could convert to HBr salt Type A after desolvation or dehydration during storage.

TABLE 25DHPLC results of HBr salt Type B (824511-10-A1)#RRTArea (%)#RRTArea (%)10.750.0950.950.0620.830.0560.970.1330.900.1371.0099.2740.930.0981.070.17

HBr Salt Type C

HBr salt Type C (824511-39-A3) was obtained by slurrying 20.3 mg HBr salt Type A (824511-29-B) in 0.5 mL DCM at RT overnight. Since no solids precipitated, the clear solution was transferred to stir at 5° C. for about two weeks and then transferred to evaporation to dryness in desiccator at RT with silica gel. The XRPD result is displayed inFIG.20Land TABLE 25E. As TGA/DSC curves inFIG.20Mshowed, a weight loss of 2.0% up to 100° C. and two endothermic signals at 142.0° C. and 208.0° C. (peak) were observed.1H NMR spectrum inFIG.20Nshowed that peak of DCM was observed. The molar ratio of DCM/API was 0.06:1 (theoretical weight=1.12 wt%). HPLC/IC results showed that the molar ratio of acid/freebase was determined as 1.0:1 and HPLC purity was 99.43 area% (FIG.20Oand TABLE 25F).

XRPD results inFIG.20Pshowed that after storage and heating HBr salt Type C to 100° C., the peak of HBr salt Type A was observed, and after heating sample to 150° C., most diffraction peaks were consistent with HBr salt Type A.1H NMR result inFIG.20Qshowed that the peak of DCM was observed. The molar ratio of DCM/API was 0.005:1 (theoretical weight=0.09 wt%). Combined with the results of heating experiment and limited solvent amount in the sample, HBr salt Type C was speculated to be a hydrate (theoretical water content for a hemi-hydrate is 1.96%) or anhydrate.

TABLE 25FHPLC results of HBr salt Type C (824511-39-A3)#RRTArea (%)#RRTArea (%)10.910.0840.980.1420.920.0551.0099.4330.960.1761.070.14

HBr Salt Type D

HBr salt Type D (824511-39-A12) was obtained by slurrying 20.0 mg HBr salt Type A (824511-29-B) in 0.5 mL THF:H2O (924:76, v/v, aw≈0.6) at RT overnight. Since no solids precipitated, the clear solution was transferred to stir at 5° C. for about two weeks and then transferred to evaporation to dryness in desiccator at RT with silica gel. The XRPD result is displayed inFIG.20Rand TABLE 25G. As shown inFIG.20S, TGA weight losses of 2.3% up to 90° C., 7.6% from 90° C. to 170° C. were observed, and three DSC endothermic signals at 70.1° C., 132.1° C. and 200.8° C. (peak) were observed.1H NMR spectrum inFIG.20Tshowed that the peak of THF was observed. The molar ratio of THF/API was 0.51:1 (theoretical weight=7.58 wt%). HPLC/IC results showed that the molar ratio of acid/freebase was determined as 1.1:1 and HPLC purity was 99.48 area% (FIG.20Uand TABLE 25H).

XRPD results inFIG.20Vshowed that after 100° C. heating of HBr salt Type D, the extra peak of HBr salt Type A was observed. After 150° C. heating, most diffraction peaks of the sample were consistent with HBr salt Type A.1H NMR result inFIG.20Wshowed that no obvious peak of THF was observed. Combined with the results of heating experiment, HBr salt Type D was speculated as a THF solvate.

TABLE 25HHPLC results of HBr salt Type D (824511-39-A12)#RRTArea (%)#RRTArea (%)10.900.1241.0099.4820.950.1851.070.1430.980.08------

Conclusion

A brief polymorph screening of 18-MC HBr salt was performed and a total of four salt forms were obtained.

18-MC tosylate Type A and Type B were first obtained as described in Example 1. Polymorph screening of 18-MC tosylate was performed to better understand polymorphism of the salt.

Tosylate material was first prepared using 18-MC freebase and used as the starting material for the polymorph screening. In the screening, different crystallization methods including temperature cycling and slurry conversion at different temperatures were used, and a total of 30 experiments were conducted. Solids from screening were isolated for XRPD testing. New forms were further characterized by TGA, DSC,1H NMR and HPLC. As the characterization and identification results showed, seven new forms (tosylate Types C, D, E, F, G, H, and I) were discovered. Results indicated that tosylate Types A, B, and C were hydrates, Types D and I were anhydrates, Types E, F, and G were solvates and Type H was a metastable form. Characterization results are summarized in TABLE 26 and XRPD patterns of different forms are displayed inFIG.21.

To summarize, a brief polymorph screen was performed and a total of nine forms of the tosylate salt were discovered.

Preparation of Tosylate Starting Material

Preparation procedure of tosylate sample was as follows: 1.0 g of freebase (824509-21-A) was weighed into a 20-mL glass vial. 4 mL of THF was added to prepare a suspension. 527.5 mg of p-toluenesulfonic acid was dissolved in 4 mL of THF. The acid solution was added into the freebase suspension dropwise with stirring. The resulting clear solution was allowed to stir at 5° C. for 4 days. Solids were isolated from suspension by centrifugation and vacuum dried at RT overnight. As a result, about 1.23 g of tosylate (824511-23-A) was obtained and it showed different XRPD pattern from tosylate Type A or B, and was assigned as tosylate Type C. After storing tosylate Type C (824511-23-A) at RT for about 2.5 month, a form change to tosylate Type B (renamed as 824511-23-A-0628) was observed (FIG.22A). The HPLC purity of tosylate Type B (824511-23-A-0628) was determined to be 99.43 area% (FIG.22Band TABLE 27A) and it was used as starting material of polymorph screening experiments.

TABLE 27AHPLC results of tosylate Type B (824511-23-A-0628)#RRTArea (%)#RRTArea (%)10.880.0540.980.0620.930.1151.0099.4330.960.1861.070.18

Approximate solubility of tosylate Type B (824511-23-A-0628) was estimated in 40 solvents to guide the solvent selection in polymorph screening of tosylate, with data summarized in TABLE 27B.

TABLE 27BApproximate solubility of tosylate Type B (824511-23-A-0628) at RT and 50° C.RT (mg/mL)50° C. (mg/mL)SolventSolubilitySolvent (v:v)SolubilitySolvent (v:v)SolubilityMeOHS>44.0ACN/toluene (4:1)S>42.0n-Butanol22.0<S<44.0EtOHS>40.0EtOH/n-hexane (3:1)S>42.0IPA/EtOAc (1:4)21.0<S<42.0CHCl3S>40.0THF/H2O (87:13)S>42.0ACN/EtOAc (1:2)19.0<S<38.0ACN22.0<S<44.0ACN/H2O (1:3)20.0<S<40.0CHCl3/n-hexane (1:1)7.3<S<22.0DCM7.3<S<22.0ACN/toluene (1:1)19.0<S<38.0Toluene/IPA (9:1)6.3<S<19.0Acetone6.7<S<20.0Acetone/n-heptane (9:1)6.3<S<19.02-MeTHF2.1<S<7.0&MEK6.7<S<20.0THF/H2O (981:19)2.1<S<7.0MIBK2.0<S<6.7&THF6.7<S<20.0EtOH/n-hexane (1:2)2.1<S<7.01,4-Dioxane/n-heptane (9:1)2.2<S<7.3IPA6.3<S<19.0CHCl3/toluene (1:1)1.9<S<6.3n-Heptane/EtOH (5:1)2.2<S<7.3EtOAcS<2.1&MeOH/H2O (1:3)1.9<S<6.3Anisole1.9<S<6.32-MeTHFS<2.1Acetone/n-heptane (4:1)S<2.0IPAcS<2.2n-HeptaneS<2.0CHCl3/toluene (1:4)S<1.9MTBES<2.1H2OS<2.0----n-HeptaneS<2.0--------TolueneS<1.9--------1,4-Dioxane/n-heptane (1:2)S<1.9&: little solid in the vial. Procedure: weigh ~2 mg solids into each 3-mL glass vial, add in corresponding solvent stepwise and sonicate or oscillate to see if solids dissolved completely. Stop adding solvent till the solids dissolves or total volume reaches 1.0 mL. The above procedure was conducted at corresponding temperature. Calculate the approximate solubility based on solvent volume.

Polymorph Screening

Using tosylate Type B (824511-23-A-0628) as the starting material, a total of 30 polymorph screening experiments were conducted via various crystallization methods. Results of polymorph screening are summarized in TABLE 27C. XRPD results showed that a total of nine forms (tosylate Types A to I) were obtained from the polymorph screening and characterization, including three hydrates (tosylate Types A, B, and C), two anhydrate (tosylate Types D and I), three solvates (tosylate Types E, F, and G) and one metastable form (tosylate Type H). Characterization data of obtained forms is summarized in TABLE 26 and the XRPD overlays of these forms are displayed inFIG.21.

TABLE 27CSummary of polymorph screening experiments of tosylateMethodNo. of ExperimentResultsTemperature Cycling5Tosylate Type BSlurry at RT20Tosylate Type B/C/B+F/B+C/B+C+extra peak/C+extra peakSlurry at 50° C.5Tosylate Type B/E/F/G/H/A+HTotal30Tosylate Type A/B/C/D, amorphous, gelNote: Tosylate Type I was observed in form identification.

Tosylate Type A was first obtained in a previous experiment. For batch 824528-06-A3-0809, it was obtained by air drying of tosylate Type H (824528-06-A3, slurry 40.2 mg tosylate (824511-23-A-0628) in 0.5 mL IPA/EtOAc (1:4, v/v) at 50° C. for 3 days) at RT for ~3 hours. The XRPD result is displayed inFIG.22Cand TABLE 27D. TGA/DSC results inFIG.22Dshowed a weight loss of 1.7% up to 100.0° C. and two endothermic peaks at 67.5° C. and 146.1° C. (peak). Using DMSO-d6as solvent,1H NMR results inFIG.22Eshowed that the peak of p-toluenesulfonic acid and EtOAc were observed. The molar ratio of p-toluenesulfonic acid/API was 1:1, the molar ratio of EtOAc/API was 0.51:1 (theoretical weight=7.79 wt%, which was higher than TGA loss and speculated to be caused by inhomogeneity of the sample). HPLC purity of the sample was determined as 99.75 area% (FIG.22Fand TABLE 27E).

VT-XRPD results inFIG.22Gshowed that after drying tosylate Type A (824528-06-A3) under N2for 30 min at 30° C., no form change was observed. After heating the sample to 100° C. under N2protection, a form change was observed, which was assigned as tosylate Type I (FIGURE and TABLE 27F). After cooling back to 30° C. under N2protection, no form change was observed for Type I. After open dish for ~3 hrs, tosylate Type I converted back to tosylate Type A with an extra peak of tosylate Type B. Thus, tosylate Type A was speculated as a hydrate and tosylate Type I was speculated as an anhydrate.

XRPD results inFIG.22Hshowed that after storing tosylate Type A at RT for ~7 days, the peak of tosylate Type B was observed. After storage for 3 weeks, the tosylate Type A sample was totally converted to tosylate Type B, which indicated Type B was possibly a hydrate or anhydrate.

TABLE 27EHPLC results of tosylate Type A (824528-06-A3)#RRTArea (%)10.960.0621.0099.7531.060.19

Tosylate Types B, C, and D

Tosylate Type C (824511-23-A) was obtained by slurrying 1.0 g freebase and 527.5 mg p-toluenesulfonic acid (charge molar ratio 1:1) in 8 mL THF at 5° C. for ~4 days. Resulting solids were isolated by centrifugation and vacuum drying at RT overnight. The XRPD result is displayed inFIG.22Jand TABLE 27G. TGA/DSC results inFIG.22Kshowed a weight loss of 4.6% up to 110° C., two endothermic signals at 93.8° C. and 128.8° C. (peak).1H NMR result inFIG.22Lshowed that the peak of p-toluenesulfonic acid and THF were observed. The molar ratio of p-toluenesulfonic acid/API was 1:1, the molar ratio of THF/API was 0.01:1 (theoretical weight=0.46 wt%). HPLC purity of the sample was determined as 99.33 area% (FIG.22Mand TABLE 27H).

VT-XRPD result inFIG.22Nshowed that after N2-drying for 30 min at 30° C., form change was observed for tosylate Type C. The new form was assigned as tosylate Type D (FIG.22Pand TABLE 271). After heating tosylate Type D to 100° C. under N2protection, no form change was observed. By heating the sample to 140° C. under N2protection, an amorphous pattern was observed. Considering limited solvent residual (much less than TGA loss), tosylate Type C was speculated to be a hydrate (theoretical water content for sesqui-hydrate is 4.76%) and tosylate Type D was an anhydrate.

XRPD result inFIG.22Oshowed that after exposing tosylate Type D to ambient conditions for ~30 min, it converted to tosylate Type B. Thus, tosylate Type B was speculated as a hydrate.

TABLE 27HHPLC results of tosylate Type C (824511-23-A)#RRTArea (%)#RRTArea (%)10.730.0550.980.0620.900.0761.0099.3330.930.0971.070.1640.960.24------

TABLE 271XRPD peak list of tosylate Type D (824511-23-A-RE_N2_60min_30.0° C.)Pos. [°2θ]Height [cts]FWHM Left [°2θ]d-spacing [Å]Rel. Int. [%]6.99711459.400.100412.63100.008.2791173.970.066910.6811.929.3038773.290.10049.5152.999.5935233.110.10049.2215.9710.0866104.330.20078.777.1511.9658202.130.13387.4013.8512.5219730.670.10047.0750.0713.7374224.690.16736.4515.4014.5331604.500.08366.1041.4216.1172668.930.10045.5045.8416.6536893.210.15065.3261.2017.0632495.190.16735.2033.9317.9862356.240.13384.9324.4118.4147511.280.16734.8235.0319.2992273.210.16734.6018.7219.9076297.000.20074.4620.3520.3050319.360.20074.3721.8821.9044404.010.11714.0627.6822.7485188.410.33463.9112.9124.0668195.890.16733.7013.4224.9065198.660.16733.5813.6125.5856107.060.26763.487.34Tosylate Type B XPRD data is shown in TABLE 27J.

Tosylate Type E

Tosylate Type E (824528-05-A9) was obtained by slurrying 19.9 mg tosylate Type B (824511-23-A-0618) in 0.5 mL 1,4-dioxane/n-heptane (9:1, v/v) at 50° C. for ~4 days. Resulting solids were isolated by centrifugation and air drying. The XRPD result is displayed inFIG.23Aand TABLE 28A. TGA/DSC results inFIG.23Bshowed a weight loss of 2.8% up to 80° C., 11.7% from 80° C. up to 130° C., and two endothermic signals at 100.2° C. and 105.4° C. (peak). Using DMSO-d6as solvent,1H NMR result inFIG.23Cshowed that the peaks of p-toluenesulfonic acid and 1,4-dioxane were observed. The molar ratio of p-toluenesulfonic acid/API was 0.9:1, the molar ratio of 1,4-dioxane/API was 0.8:1 (theoretical weight=11.84 wt%). HPLC purity of the sample was determined as 99.85 area% (FIG.23Dand TABLE 28B).

XRPD results inFIG.23Eshowed that after heating tosylate Type E to 80° C., cooling back to RT under N2 (10° C./min heating and cooling rate) and re-exposing to ambient conditions for XRPD test, no form change was observed. After heating tosylate Type E to 101° C. and cooling back to RT, a sample with weak crystallinity and similar to tosylate Type B was obtained.1H NMR result inFIG.23Fshowed that the amount of 1,4-dioxane decreased obviously (molar ratio of 1,4-Dioxane/API was 0.05:1, theoretical weight=0.88 wt%). Combined with the results of heating experiments, tosylate Type E was speculated as a 1,4-dioxane solvate.

TABLE 28BHPLC results of tosylate Type E (824528-05-A9)#RRTArea (%)10.960.0721.0099.8531.060.09

Tosylate Type F

Tosylate Type F (824528-06-B1) was prepared by slurrying 40.3 mg tosylate Type B (824511-23-A-0618) in 0.5 mL CHCl3/n-hexane (1:1, v/v) at 50° C. for ~6 days. Resulting solids were isolated by centrifugation and air drying. The XRPD results are displayed inFIG.23G,FIG.23Hand TABLE 28C. TGA/DSC results inFIG.23Ishowed a weight loss of 17.4% up to 100° C., three endothermic signals at 74.1° C., 96.8° C. and 122.5° C. (peak). Using DMSO-d6as solvent,1H NMR result inFIG.23Jshowed that the peak of p-toluenesulfonic acid and CHCl3were observed. The molar ratio of p-toluenesulfonic acid/API was 1:1, the molar ratio of CHCl3/API was 0.41:1 (theoretical weight=8.22 wt%). HPLC purity of the sample was determined as 99.52 area% (FIG.23Kand TABLE 28C).

XRPD results inFIG.23Lshowed that after heating tosylate Type F to 90° C., cooling back to RT under N2 (10° C./min heating and cooling rate) and re-exposing to ambient conditions for XRPD test, form change to tosylate Type B was observed.1H NMR result inFIG.23Mshowed that no obvious peak of CHCl3was observed. Combined with the results of heating experiment, tosylate Type F was speculated as a CHCl3solvate.

Tosylate Type G

Tosylate Type G (824528-06-A1) was obtained by slurrying 50.2 mg tosylate Type B (824511-23-A-0618) in 0.5 mL anisole at 50° C. for ~3 days. Resulting solids were isolated by centrifugation and air drying. The XRPD results are displayed inFIG.23N,FIG.23Oand TABLE 28E. TGA/DSC results inFIG.23Pshowed a weight loss of 1.2% up to 70° C., 7.1% from 70° C. up to 120° C. and one endotherm ic signal at 107.9° C. (peak). Using DMSO-d6as solvent,1H NMR result inFIG.23Qshowed that the peak of p-toluenesulfonic acid and anisole were observed. The molar ratio of p-toluenesulfonic acid/API was 0.9:1, the molar ratio of anisole/API was 0.47:1 (theoretical weight=8.70 wt%). HPLC purity of the sample was determined as 99.63 area% (FIG.23Rand TABLE 28F).

XRPD results inFIG.23Sshowed that after heating tosylate Type G to 80° C., cooling back to RT under N2 (10° C./min heating and cooling rate) and re-exposing to ambient conditions for XRPD test, no form change was observed. After heating tosylate Type G to 90° C. and 100° C., crystallinity of the sample decreased significantly (90° C.), and amorphous was obtained (100° C.).1H NMR result inFIG.23Tshowed that along with the temperature increased, the solvent content decreased when the crystallinity of the sample decreased. The molar ratio of anisole/API was decreased to 0.26:1 (theoretical weight=5.10 wt%) after 100° C. heating. Tosylate Type G was speculated as an anisole solvate.

Metastable Form

Tosylate Type H (824528-05-A12) was obtained by slurrying 23.5 mg tosylate Type B (824511-23-A-0618) in 0.5 mL IPA/EtOAc (1:4, v/v) at 50° C. for ~4 days. Resulting solids were isolated by centrifugation and air drying. The XRPD result is displayed inFIG.24Aand TABLE 29. The XRPD overlay inFIG.24Bshowed that after air drying of tosylate Type H (824528-06-A3) for ~3 hours, a form change to tosylate Type A was observed, which indicated that tosylate Type H was a metastable form and no more characterization data were collected.

Conclusion

A brief polymorph screening of 18-MC tosylate was performed and a total of nine salt forms were obtained.

18-MC besylate Type A and Type B were obtained as described in Example 1. Polymorph screening of 18-MC besylate was performed to better understand polymorphism of the salt.

The besylate material was first prepared using 18-MC freebase and then used as starting material for polymorph screening. In the screen, different crystallization methods, including temperature cycling and slurry conversion at different temperatures, were used, and a total of 30 experiments were conducted. Solids from screening were isolated for X-ray powder diffraction (XRPD) testing. From the results of characterization and form identification, a total of three besylate forms were obtained, including two anhydrates, besylate Types B and C, and one hydrate, besylate Type A. Characterization results are summarized in TABLE 30 and XRPD patterns of different forms are displayed inFIG.25.

To summarize, a brief polymorph screen was performed and a total of three forms of besylate were discovered.

Preparation of Besylate Starting Material

Preparation procedure of the besylate (824511-44-C2) was as follows: 1.0 g of freebase (824509-24-A) was weighed into a 20-mL glass vial along with 436.6 mg of benzenesulfonic acid. Then, 6 mL of IPA was added to the vial to produce a suspension which was slurried at RT for ~7 days. The resulting sample was centrifuged and vacuum dried at RT for 6 hours. As the results (FIGURE ) showed, about 1.15 g of besylate Type B (824511-44-C2) was obtained.

Approximate solubility of besylate Type B (824511-44-C2) was estimated in 38 solvents to guide the solvent selection in polymorph screening of besylate, with results shown in TABLE 31.

TABLE 31Approximate solubility of besylate Type B (824511-44-C2) at RT and 50° C.RT (mg/mL)50° C. (mg/mL)SolventSolubilitySolvent (v:v)SolubilitySolvent (v:v)SolubilityMeOHS>44.0THF/H2O (87:13, aW0.8)S>42.0ACN/EtOAc (1:2)20.0<S<40.0DCMS>40.0IPA/H2O (847:153, aw0.8)S>40.0IPA/EtOAc (1:2)7.0<S<21.0CHCl3S>40.0THF/H2O (981:19, aw0.2)S>40.0IPAc/acetone (1:1)2.2<S<7.3ACNS>40.0DCM/EtOAc (1:1)7.7<S<23.0n-Heptane /EtOH (1:1)2.2<S<7.3Acetone21.0<S<42.0EtOH/n-hexane (2:1)7.0<S<21.01,4-Dioxane/ n-heptane (9:1)2.1<S<7.0EtOH20.0<S<40.0ACN/toluene (1:4)6.3<S<19.0MTBE/CHCl3(1:1)2.0<S<6.7THF7.0<S<21.0MeOH/H2O (2:1, aw0.6)2.0<S<6.7Anisole2.0<S<6.7MEK6.3<S<19.02-MeTHF/DCM (4:1)2.0<S<6.7Toluene/IPA (9:1)2.0<S<6.7IPA2.0<S<6.7IPA/H2O (982:18, aw0.2)2.0<S<6.7CHCl3/ n-hexane (1:1)1.9<S<6.3EtOAcS<2.0CHCl3/toluene (1:1)2.0<S<6.7MIBK1.9<S<6.3n-HeptaneS<2.0Acetone/ n-heptane (4:1)1.9<S<6.3n-HeptaneS<2.0H2OS<2.0ACN/H2O (1:2, aw0.9)S<1.9*2-MeTHFS<1.9*2-MeTHFS<1.9----TolueneS<1.9*: Little solids in the vial.Procedure: weigh ~2 mg solids into each 3-mL glass vial, add in corresponding solvent stepwise and sonicate or oscillate to see if solids dissolved completely. Stop adding solvent till the solids dissolves or total volume reaches 1.0 mL. Calculate the approximate solubility based on solvent volume.Procedure: weigh ~2 mg solids into each 3-mL glass vial, add in corresponding solvent stepwise and sonicate or oscillate to see if solids dissolved completely. Stop adding solvent till the solids dissolves or total volume reaches 1.0 mL. Calculate the approximate solubility based on solvent volume.

Polymorph Screening

Using besylate Type B (824511-44-C2) as the starting material, a total of 30 polymorph screening experiments were conducted via various crystallization methods. Results of polymorph screening is summarized in TABLE 32A. XRPD results showed that a total of 3 forms (besylate Types A, B, and C) were obtained from the screening and characterization studies, including two anhydrates (besylate Types B and C) and one hydrate (besylate Type A). Characterization data of obtained forms is summarized in TABLE 30 and the XRPD overlays of these forms are displayed inFIG.25.

TABLE 32ASummary of polymorph screening experiments of besylateMethodNo. of ExperimentResultsTemperature cycling5Besylate Type A, Type BSlurry at RT13Besylate Type A, Type B, Type A+BSlurry at 50° C.12Besylate Type A, Type B, Type A+B+extra peakTotal30Besylate Type A, Type B, Type A+B, Type A+B+extra peakNote: Besylate Type C was discovered in form identification of besylate Type A.

Besylate Type A and C

Besylate Type A (824511-35-A1) was obtained by slurrying 40.2 mg freebase and 17.4 mg benzenesulfonic acid (charge molar ratio of acid to freebase was 1:1) in 0.5 mL DCM/EtOAc (1:1, v/v) at RT for ~2 days. Resulting solids were isolated by centrifugation and vacuum drying at RT overnight. The XRPD results are displayed inFIG.26AandFIG.26B. Thermogravimetric analysis (TGA)/differential scanning calorimetry (DSC) curves inFIG.26Cshowed a weight loss of 1.1% up to 90.0° C. and a weight loss of 3.3% from 90° C. to 130° C., and two endothermic DSC signals at 117.4° C. and 131.8° C. (peak). Using DMSO-d6as the solvent, proton nuclear magnetic resonance (1H NMR) results inFIG.26Dshowed that the peak of benzenesulfonic acid was observed. The molar ratio of acid/API was 1.0:1. No obvious solvent residual was detected. High performance liquid chromatography (HPLC) purity was 99.71 area% (FIG.26Eand TABLE 32B).

VT-XRPD test was performed using another batch of besylate Type A (824529-04-A5). As the results displayed inFIG.26F, after heating besylate Type A to 100° C. and cooling back to 30° C. under N2protection, a new form was observed and assigned as besylate Type C (FIG.26Gand TABLE 32C). After exposure to ambient condition for ~30 min besylate Type C converted back to besylate Type A. Thus, besylate Type A was speculated as a hydrate (theoretical water content for monohydrate is 3.73%) and besylate Type C was speculated as an anhydrate.

TABLE 32BHPLC results of besylate Type A (824511-35-A1)#RRTArea (%)10.910.0520.970.0531.0099.7141.070.18

Besylate Type B

Besylate Type B (824511-44-C2) was prepared by slurrying 1.0 g freebase and 436.6 mg benzenesulfonic acid (charge molar ratio of acid to freebase was 1:1) in 6.0 mL IPA at RT for ~7 days. Resulting solids were isolated by centrifugation and vacuum drying at RT for about 6 hrs. The XRPD result of besylate Type B (824511-44-C2) is shown inFIG.26H, FIGURE and TABLE 32D. TGA/DSC curves inFIG.26Jshowed a weight loss of 2.4% up to 150.0° C. and one endothermic peak at 181.1° C. (peak).1H NMR result inFIG.26Kshowed that the peak of benzenesulfonic acid and IPA were observed. The molar ratio of benzenesulfonic acid/API was 1:1, the molar ratio of IPA/API was 0.22:1 (theoretical weight=2.50 wt%). HPLC purity was 99.73 area% (FIG.26Land TABLE 32E).

VT-XRPD results inFIG.26Mshowed that after drying besylate Type B (824511-44-C2) under N2 for 20 min at 30° C., or heating sample to 100° C. and cooling back to 30° C. under N2protection, no obvious form change was observed, indicating besylate Type B was an anhydrate.

Conclusion

A brief polymorph screen of 18-MC besylate was performed and a total of three polymorphs were obtained.

Throughout this application, various publications, including United States patents, are referenced by author and year and patents by number. Full citations for the publications are listed below. The disclosures of these publications and patents in their entireties are hereby incorporated by reference into this application in order to more fully describe the state of the art to which this invention pertains.