Fumagilol derivatives and their polymorphs

Abstract

To provide salts and polymorphs of aminoalkylfumagillol carbamates (e.g., fumagill-6-yl N-(trans-4-aminocyclohexyl)carbamate benzenesulfonic acid salts and fumagill-6-yl N-(trans-4-aminocyclohexyl)carbamate hydroxynaphthoic acid salts).SOLUTION: The polymorphs are characterized by X-ray powder diffraction, differential scanning calorimetry, thermogravimetric analysis, and other methods. The polymorphs and salts can be used as intermediates in production of fumagillol derivatives (e.g., polymer-conjugated fumagillol derivatives) and used for treatment of various diseases and conditions such as cancer.SELECTED DRAWING: None

Description

[Background technology]

[0001] Cross-reference of related applications

[0002] This application claims priority to U.S. Provisional Patent Application No. 62 / 265,675, filed on 10 December 2015.

[0003] This application relates to salts and polymorphs of aminoalkylfumagilol carbamates (e.g., fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamate benzenesulfonate and fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamate hydroxynaphthoethate). The salts and polymorphs of this application are useful as stable intermediates in the production of fumagilol derivatives (e.g., polymer-conjugated fumagilol derivatives) or as therapeutic agents.

[0004] Fumagiline is a low-molecular-weight substance used as an antimicrobial and antiprotozoan. The clinical applications of fumagiline and its derivatives have been limited by toxicity. The formation of fumagiline polymer conjugates for therapeutic purposes requires the presence of one or more stable, pure fumagiline-derived intermediates that do not readily self-condense or hydrolyze (e.g., by the reaction of amine-modified fumagilol with an epoxide on fumagilol). Furthermore, impurities covalently bonded to the polymer backbone are difficult to remove. [Overview of the project]

[0005] This disclosure relates to pure and stable fumagillol derivatives (e.g., formulas I and II). These derivatives can be used as synthetic intermediates for producing polymer-bound fumagillol conjugates. Alternatively, the derivatives can be used directly as therapeutic agents.

[0006] This application relates to salts of aminoalkylfumagilol carbamates, for example, fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamate benzenesulfonate, (1R,4R)-4-(((((3R,4S,5S,6R)-5-methoxy-4-((2R,3R)-2-methyl-3-(3-methylbuta-2-en-1-yl)oxiran-2-yl)-1-oxaspiro[2.5]octan-6-yl)oxy)carbonyl)amino)cyclohexane-1-aminiumbenzenesulfonate, and fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamate hydroxynaphthoate, (1R,4R)-4-(((((3R,4S,5S,6R)-5-methoxy-4- The subject is ((2R,3R)-2-methyl-3-(3-methylbuta-2-en-1-yl)oxiran-2-yl)-1-oxaspiro[2.5]octan-6-yl)oxy)carbonyl)amino)cyclohexane-1-aminium-1-hydroxy-2-naphthoate, also known as this disclosure. The disclosure also provides stable and pure crystalline polymorphs of both salts. The salts and polymorphs provided herein are stable and can be used as intermediates in the production of fumagilol derivatives (e.g., polymer-conjugated fumagilol derivatives). These derivatives are useful as therapeutic agents in the treatment of many diseases. Furthermore, the aforementioned salts and polymorphs can be used directly as therapeutic agents.

[0007] In one aspect, the present disclosure relates to fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate (Formula I): [ka] Write it down.

[0008] In one embodiment, the present disclosure provides fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate (Formula I) (e.g., Form I, Form II, Form III and Form IV).

[0009] In one embodiment, the disclosure provides a fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate polymorph (morph II) characterized by an X-ray powder diffraction pattern using Cu Kα irradiation, including peaks at approximately 6.0, 9.0, and 18.0°2θ. In one or more embodiments, the polymorph of morph II is characterized by the onset of an exothermic event at approximately 178°C and a peak at approximately 188°C, as measured by differential scanning calorimetry. In one or more embodiments, the polymorph of morph II is characterized by the onset of an exothermic event at approximately 181°C and a peak at approximately 189°C, as measured by thermogravimetric analysis / differential thermal analysis (TG / DTA). In one or more embodiments, morph II is characterized by an X-ray powder diffraction pattern substantially similar to that shown in Figure 5A or 6A.

[0010] In one embodiment, the disclosure provides a fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate polymorph (Form I) characterized by an X-ray powder diffraction pattern using Cu Kα irradiation, including peaks at approximately 6.1, 8.6, and 16.4°2θ. In some embodiments, Form I is characterized by an X-ray powder diffraction pattern substantially similar to that shown in Figure 5B or 6B.

[0011] In one embodiment, the disclosure provides a fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate polymorph (Form III) characterized by an X-ray powder diffraction pattern using Cu Kα irradiation, including peaks at approximately 8.3, 12.8, and 15.7°2θ. In some embodiments, Form III is characterized by an X-ray powder diffraction pattern substantially similar to that shown in Figure 5C or 6C.

[0012] In one embodiment, the disclosure provides a fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate polymorph (Form IV) characterized by an X-ray powder diffraction pattern using Cu Kα irradiation, including peaks at approximately 5.1, 7.3 and 14.0 °2θ. In some embodiments, Form IV is characterized by an X-ray powder diffraction pattern substantially similar to that shown in Figure 5D or 6D.

[0013] In another embodiment, the present disclosure relates to fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamate hydroxynaphthoethate (Formula II): [ka] Write it down.

[0014] In another embodiment, the disclosure describes fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamate hydroxynaphthoethate (Formula II) (e.g., Form A).

[0015] In one embodiment, the disclosure provides a fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamate hydroxynaphthoate polymorph (morph A) characterized by an X-ray powder diffraction pattern using Cu Kα irradiation, including peaks at approximately 5.6, 8.9, and 15.4 °2θ. In one or more embodiments, the polymorph of morph A is characterized by an endothermic onset at approximately 182 °C and a peak at approximately 186 °C when measured by differential scanning calorimetry. In some embodiments, the polymorph of morph A is characterized by an endothermic onset at approximately 181 °C with a peak at approximately 186 °C and exothermic at approximately 191 °C (peak) when measured by thermogravimetric / differential thermal analysis (TG / DTA).

[0016] In another embodiment, this disclosure is as follows: (i) Step 1: Dissolve fumaril-6-yl N-(trans-4-aminocyclohexyl)carbamate benzenesulfonate in methanol, optionally containing MTBE, to form a solution; (ii) Step 2: Optionally, add seed crystals of fumaril-6-yl N-(trans-4-aminocyclohexyl)carbamate benzenesulfonate (e.g., about 1% by weight) to the solution; (iii) Step 3: Optionally, separate the crystallized fumaril-6-yl N-(trans-4-aminocyclohexyl)carbamate benzenesulfonate from the solution. A method for preparing a polymorph of fumaril-6-yl N-(trans-4-aminocyclohexyl)carbamate benzenesulfonate (Formula I) is provided, which includes the above steps.

[0017] In some embodiments, the method optionally includes, after Step 1 but before Step 2, Step 1A and / or 1B: Step 1A: Cool the fumaril-6-yl N-(trans-4-aminocyclohexyl)carbamate benzenesulfonate solution obtained from the previous step; Step 1B: Add additional MTBE to the fumaril-6-yl N-(trans-4-aminocyclohexyl)carbamate benzenesulfonate solution obtained from the previous step; and further includes these steps.

[0018] In some embodiments, the method optionally includes, after Step 2 but before Step 3, Step 2A and / or 2B: Step 2A: Cool the fumaril-6-yl N-(trans-4-aminocyclohexyl)carbamate benzenesulfonate solution obtained from the previous step; Step 2B: Add MTBE to the fumaril-6-yl N-(trans-4-aminocyclohexyl)carbamate benzenesulfonate solution obtained from the previous step; and further includes these steps.

[0019] In one or more embodiments, the method includes steps 1, 2, and 3. In some embodiments, the method includes steps 1, 1A, 2, and 3. In some embodiments, the method includes steps 1, 1B, 2, and 3. In some embodiments, the method includes steps 1, 2, 2A, and 3. In some embodiments, the method includes steps 1, 2, 2B, and 3. In some embodiments, the method includes steps 1, 1A, 2, 2A, and 3. In some embodiments, the method includes steps 1, 1A, 2, 2B, and 3. In some embodiments, the method includes steps 1, 1B, 2, 2A, and 3. In some embodiments, the method includes steps 1, 1B, 2, 2B, and 3. In some embodiments, the method includes steps 1, 1A, 1B, 2, and 3. In some embodiments, the method includes steps 1, 1B, 1A, 2, and 3. In some embodiments, the method includes steps 1, 1A, 1B, 2, 2A, and 3. In some embodiments, the method includes steps 1, 1A, 1B, 2, 2B, and 3. In some embodiments, the method includes steps 1, 1B, 1A, 2, 2A, and 3. In some embodiments, the method includes steps 1, 1B, 1A, 2, 2B, and 3. In some embodiments, the method includes steps 1, 2A, 2B, and 3. In some embodiments, the method includes steps 1, 1A, 1B, 2, 2A, 2B, and 3. In some embodiments, the method includes steps 1, 1A, 1B, 2, 2B, 2A, and 3. In some embodiments, the method includes steps 1, 1A, 1B, 2, 2B, 2A, and 3. In some embodiments, the method includes steps 1, 1B, 2, 2A, 2B, and 3. In some embodiments, the method includes steps 1, 1B, 2, 2B, 2A, and 3.In some embodiments, the method includes Step 1, Step 1A, Step 1B, Step 2, Step 2A, Step 2B, and Step 3. In some embodiments, the method includes Step 1, Step 1A, Step 1B, Step 2, Step 2B, Step 2A, and Step 3. In some embodiments, the method includes Step 1, Step 1B, Step 1A, Step 2, Step 2A, Step 2B, and Step 3. In some embodiments, the method includes Step 1, Step 1B, Step 1A, Step 2, Step 2B, Step 2A, and Step 3.

[0020] In one or more embodiments, the present disclosure is as follows: (i) Step 1': Add benzenesulfonic acid (e.g., a solution of benzenesulfonic acid in tert-butyl methyl ether or ethyl acetate) to a solution of fumagil-6-yl N-(trans-4-aminocyclohexyl) carbamate benzenesulfonate in a solvent (e.g., tert-butyl methyl ether, methanol, a combination of tert-butyl methyl ether and methanol,or ethyl acetate). This addition can form a precipitate that is later isolated (e.g., by filtration). The precipitate containing fumagil-6-yl N-(trans-4-aminocyclohexyl) carbamate benzenesulfonate can be redissolved in a solvent later for further recrystallization of Step 1 above. Provided is a method for reaction crystallization of a polymorph of fumagil-6-yl N-(trans-4-aminocyclohexyl) carbamate benzenesulfonate, including the above.

[0021] In some embodiments, the step further includes one or more steps selected from the above Step 1A, Step 1B, Step 2, Step 2A, Step 2B, and Step 3.

[0022] In some embodiments, the method includes steps 1', 1A, and 3. In some embodiments, the method includes steps 1', 1A, 2, and 3. In some embodiments, the method includes steps 1', 2, 2A, and 3. In some embodiments, the method includes steps 1', 1B, 2, and 3. In some embodiments, the method includes steps 1', 2, 2B, and 3.

[0023] In some embodiments, the method includes steps 1', 2, and 3. In some embodiments, the method includes steps 1', 1A, 2, and 3. In some embodiments, the method includes steps 1', 1B, 2, and 3. In some embodiments, the method includes steps 1', 2, 2A, and 3. In some embodiments, the method includes steps 1', 2, 2B, and 3. In some embodiments, the method includes steps 1', 1A, 2, 2A, and 3. In some embodiments, the method includes steps 1', 1A, 2, 2B, and 3. In some embodiments, the method includes steps 1', 1B, 2, 2A, and 3. In some embodiments, the method includes steps 1', 1B, 2, 2B, and 3. In some embodiments, the method includes steps 1', 1B, 2, 2B, and 3. In some embodiments, the method includes steps 1', 1B, 1A, 2, and 3. In some embodiments, the method includes steps 1', 1A, 1B, 2, 2A, and 3. In some embodiments, the method includes steps 1', 1A, 1B, 2, 2B, and 3. In some embodiments, the method includes steps 1', 1B, 1A, 2, 2A, and 3. In some embodiments, the method includes steps 1', 1B, 1A, 2, 2B, and 3. In some embodiments, the method includes steps 1', 2, 2A, 2B, and 3. In some embodiments, the method includes steps 1', 2, 2B, and 3. In some embodiments, the method includes steps 1', 2, 2B, and 3. In some embodiments, the method includes steps 1', 1A, 2, 2B, 2A, and 3.In some embodiments, the method includes steps 1', 1B, 2, 2B, 2A, and 3. In some embodiments, the method includes steps 1', 1A, 1B, 2, 2A, 2B, and 3. In some embodiments, the method includes steps 1', 1A, 1B, 2, 2B, 2A, and 3. In some embodiments, the method includes steps 1', 1B, 1A, 2, 2A, 2B, and 3. In some embodiments, the method includes steps 1', 1B, 1A, 2, 2B, 2A, and 3.

[0024] In one or more embodiments, benzenesulfonic acid is added to a solution of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamate as a solution in a suitable solvent (e.g., MTBE, methanol, or a combination thereof).

[0025] In one or more embodiments, adding one or a combination of steps 1A, 1B, 2A, and 2B can improve the yield and / or purity of the resulting polymorphs.

[0026] In one embodiment, this disclosure relates to Formula III: [ka] (In the formula, AA is a naturally occurring or unnatural amino acid, x is an integer between 1 and approximately 450, y is an integer between 1 and approximately 30, and n is an integer between 1 and approximately 50.) The present invention provides a method for producing the compound shown by formula III. For example, the compound shown by formula III is compound 1: [ka] That's fine.

[0027] In some embodiments, the method involves a compound represented by formula (I) or formula (II): [ka] [ka] The compound represented by formula (IV): [ka] (wherein x, y, and n are as defined above, and LG is a leaving group (e.g., a para-nitrophenyl group), This includes making contact with it.

[0028] In another embodiment, the present disclosure provides the crystallization of compounds represented by formula I or formula II from ethyl acetate.

[0029] In other embodiments, the disclosure provides the use of compounds represented by formula I or formula II for the treatment of diseases.

[0030] In one or more embodiments, the Disclosure provides compositions comprising a compound represented by formula I or formula II in high purity.

[0031] In one or more embodiments, the disclosure provides compositions comprising a polymorph of formula I and a pharmaceutically acceptable carrier. In one or more embodiments, the disclosure provides compositions comprising polymorph II of formula I and a pharmaceutically acceptable carrier.

[0032] In one or more embodiments, the disclosure provides compositions comprising a polymorph of formula II and a pharmaceutically acceptable carrier. In one or more embodiments, the disclosure provides compositions comprising polymorph A of formula II and a pharmaceutically acceptable carrier.

[0033] The polymorphs of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamate benzenesulfonate and fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamate hydroxynaphthoethate provided herein are more stable than the corresponding free base, fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamate, and can be isolated with higher purity than the free base. Furthermore, the polymorphs (e.g., form II) are more stable than certain other salts of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamate, including hemitartrate. Due to their increased purity and improved stability, the salts and polymorphs provided herein can be used as intermediates in the synthesis of polymer conjugate fumagilol derivatives. The polymer conjugates can then be used as therapeutic agents for the treatment of diseases such as MetAP2-related diseases. Furthermore, the salts and polymorphs can be used as therapeutic agents.

[0034] In another embodiment, the disclosure provides the use of compounds of formula I or formula II, or polymorphs thereof, to inhibit or reduce the activity of MetAP2. In one or more embodiments, the activity of MetAP2 inhibitors may influence the onset and progression of diseases such as cancer.

[0035] In one aspect, the disclosure provides a method for treating a disease, comprising administering an effective amount of a compound of formula I, formula II, or formula III to a subject in need thereof.

[0036] In one aspect, the disclosure provides the use of a compound of formula I, formula II, or formula III in the manufacture of a pharmaceutical product for the treatment of a disease.

[0037] In one aspect, the present disclosure provides compounds of formula I, formula II, or formula III for the treatment of a disease.

[0038] The aforementioned disease may be, for example, cancer or a metabolic disease.

[0039] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as generally understood by those skilled in the art to which this application belongs. In this specification, the singular form includes the plural form unless otherwise explicitly indicated by the context. Similar or equivalent methods and materials may be used in the implementation or testing of this application, but suitable methods and materials are described below. All publications, patent applications, patents, and other references referenced herein are incorporated herein by reference. References cited herein are not considered prior art to this application. In case of any conflict, this specification, including definitions, shall prevail. Furthermore, materials, methods, and examples are illustrative and not intended to limit the scope. Further features and advantages of the technology disclosed herein will become apparent to those skilled in the art by reading the detailed description below. [Brief explanation of the drawing]

[0040] [Figure 1A] Figure 1A shows the XRPD spectrum (top) of a sample of the morph II polymorph of formula I (fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamate benzenesulfonate) along with the reference spectrum (bottom).

[0041] [Figure 1B] Figure 1B shows the DSC curve for a sample of morphological polymorph I.

[0042] [Figure 1C] Figure 1C shows the TG / DTA plot of a morphology II polymorph sample of formula I. The TG plot is the upper line, and the DTA plot is the lower line.

[0043] [Figure 2A] Figure 2A, similar to Figure 1A, shows the XRPD spectrum (top) of a morphology II polymorph sample of formula I, along with the reference spectrum (bottom).

[0044] [Figure 2B]Figure 2B shows the DSC curve for a sample of the morphological polymorph I.

[0045] [Figure 2C] Figure 2C shows the TG / DTA plot of a morphology II polymorph sample of formula I. The TG plot is the upper line, and the DTA plot is the lower line.

[0046] [Figure 3A-3C] Figure 3A shows the XRPD spectrum of a first sample of the morph II polymorph of formula I, prepared by the cooling method described herein as Cooling-1. Figure 3B shows the XRPD spectrum of a second sample of the morph II polymorph of formula I, prepared by the cooling method described herein as Cooling-1. Figure 3C shows the XRPD spectrum of a third sample of the morph II polymorph of formula I, prepared by the cooling method described herein as Cooling-1.

[0047] [Figure 4A-4C] Figure 4A is the XRPD spectrum of the first sample of the polymorph II of formula I, prepared by the poor solvent method described herein as Antisol-1. Figure 4B is the XRPD spectrum of the second sample of the polymorph II of formula I, prepared by the poor solvent method described herein as Antisol-1. Figure 4C is the XRPD spectrum of the third sample of the polymorph II of formula I, prepared by the poor solvent method described herein as Antisol-1.

[0048] [Figures 5A-5D] Figure 5A shows the XRPD spectrum of a sample of the morphology II polymorph of formula I. Figure 5B shows the XRPD spectrum of a sample of the morphology I polymorph of formula I. Figure 5C shows the XRPD spectrum of a sample of the morphology III polymorph of formula I. Figure 5D shows the XRPD spectrum of a sample of the morphology IV polymorph of formula I.

[0049] [Figure 6A]Figure 6A shows the XRPD spectrum of a morphology II polymorph sample of formula I, with selected peaks. The list of peaks is shown in Tables 2, 3, 4, 5, 6, and 7.

[0050] [Figure 6B] Figure 6B shows the XRPD spectrum of a morphology I polymorph sample of formula I, with selected peaks. A list of peaks is shown in Table 1.

[0051] [Figure 6C] Figure 6C shows the XRPD spectrum of a morphology III polymorph sample of formula I, with selected peaks. A list of peaks is shown in Table 9.

[0052] [Figure 6D] Figure 6D shows the XRPD spectrum of a morphological IV polymorph sample of formula I, with selected peaks. A list of peaks is shown in Table 10.

[0053] [Figure 7] Figure 7 shows the ¹H NMR spectrum of Equation I, along with the impurities in the solvent.

[0054] [Figure 8] Figure 8 shows a single molecule of the morphological II polymorph of formula I, as determined by SXRD analysis.

[0055] [Figure 9] Figure 9 shows the HPLC analysis of formula I.

[0056] [Figure 10A-10D]Figure 10A shows the XRPD spectrum of a morph A polymorph sample of formula II (fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamate hydroxynaphthoethate) crystallized from acetonitrile. Figure 10B shows the XRPD spectrum of a morph A polymorph sample of formula II crystallized from ethyl acetate. Figure 10C shows the XRPD spectrum of a morph A polymorph sample of formula II crystallized from methyl ethyl ketone. Figure 10D shows the XRPD spectrum of a morph A polymorph sample of formula II crystallized from tetrahydrofuran.

[0057] [Figure 10E] Figure 10E shows the XRPD spectrum of a sample of the morphological A polymorph of formula II, with selected peaks.

[0058] [Figure 11] Figure 11 shows the DSC curve for a sample of the morphological A polymorph of Equation II.

[0059] [Figure 12] Figure 12 shows the TG / DTA plot of the morphology II polymorph sample of formula I. The TG plot is a line that slopes downward from left to right, and the DTA plot is a line that slopes upward from left to right.

[0060] [Figure 13] Figure 13 shows the 1H NMR spectrum of Equation II.

[0061] [Figure 14A] Figure 14A shows the HPLC analysis of hydroxynaphthoate (Formula II) of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamate obtained from the solid compound. The same plot is shown in low resolution (top panel) and high resolution (bottom panel).

[0062] [Figure 14B]Figure 14B shows the HPLC analysis of hydroxynaphthoate (Formula II) of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamate obtained from the solid compound. The same plot is shown in low resolution (top panel) and high resolution (bottom panel).

[0063] [Figure 15A] Figure 15A shows the effect of Equation I on B-cell lymphoma cells as a concentration function.

[0064] [Figure 15B] Figure 15B shows the effect of Equation I on mantle cell lymphoma cells as a concentration function.

[0065] [Figure 15C] Figure 15C shows the effect of Equation I on diffuse large B-cell lymphoma cells as a concentration function.

[0066] [Figure 15D] Figure 15D shows the effect of formula I on myeloma cells as a concentration function. Detailed description of the invention Polymorph of this application

[0067] This disclosure relates to salts of aminoalkylfumagilol carbamates (e.g., benzenesulfonates and hydroxynaphthoates) and their polymorphs. Specifically, this disclosure describes crystalline polymorphs of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamate benzenesulfonate (Formula I) and fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamate hydroxynaphthoates (Formula II). The crystalline polymorph of Formula I may be type I, type II, type III, or type IV. In some embodiments, the polymorph is type II. In some embodiments, the polymorph is a solvate. This disclosure also provides polymorph A of Formula II. The crystalline polymorphs described herein can be used as intermediates in the synthesis of polymer conjugate derivatives of fumagilol or as therapeutic agents.

[0068] In one or more embodiments, the Disclosure provides compositions comprising a high-purity compound of formula I or formula II. In some embodiments, the compositions may further comprise a pharmaceutical carrier. In some embodiments, the compound of formula I or II is of high purity, for example, having a purity higher than about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, 99.5%, about 99.9%, or about 99.99%. Compounds of formula I or II (e.g., fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamate or benzenesulfonate of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamate hydroxynaphthoethate) can be obtained by crystallization in high purity (e.g., higher than about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, 99.5%, about 99.9%, or about 99.99%). While not intended to imply any constraints on theory, for free bases (i.e., fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamate free base) or certain other salts of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamate, achieving high-purity compounds such as formula I or II is difficult due to a considerable amount of impurities, such as products of the autocondensation of the free base.

[0069] In some embodiments, the polymorphs or formula I or II described herein include a solvate. The solvent present in the solvate may be, for example, an organic solvent in which the polymorph is crystallized, or water. In some embodiments, the solvent may be water or MTBE. In some embodiments, the polymorphs described herein include less than 10% of a solvent (e.g., less than 5%, less than 4%, less than 3%, less than 2%, or less than 1%) such as MTBE. In some embodiments, the polymorph includes less than 1% of water (e.g., less than 0.5%, less than 0.4%, less than 0.3%, less than 0.2%, or less than 0.1%). In addition to MTBE, the solvent may be an organic solvent used in a recrystallization process, such as ethyl acetate.

[0070] As defined herein, “XRPD” or “XPD” is understood to mean powder X-ray diffraction. The abbreviation “DSC” is understood to mean differential scanning calorimetry. The abbreviation “TG” is understood to mean thermogravimetric analysis. The abbreviation “DTA” is understood to mean differential thermal analysis, and the abbreviation “TG / DTA” is understood to mean thermogravimetric analysis / differential thermal analysis.

[0071] As defined herein, "GMP" means Good Manufacturing Practice, which is a standard for manufacturing and quality control.

[0072] As used herein, "MTBE" means methyl tert-butyl ether, also known as tBME.

[0073] As defined herein, “pure” is understood to mean that a compound is homogeneous in its chemical composition. A pure compound is understood to not contain large amounts of molecules of another chemical composition, such as less than 5%, less than 4%, less than 3%, less than 2%, or less than 1%. In some cases, purity is measured excluding the solvent (e.g., organic solvents such as MTBE and ethyl acetate, and inorganic solvents such as water).

[0074] As defined herein, “stable” or “stable” refers to a compound’s ability to remain pure for a certain period of time. A stable compound may maintain its purity (e.g., without molecules of undesirable chemical formulas) despite long-term storage (e.g., more than one month, more than six months, or more than one year). A stable compound may also remain pure despite conditions such as high temperature or high humidity. Formula I: Fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate Form I, Formula I

[0075] Form I of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate (Formula I) can be crystallized from acetonitrile, for example, at about 50°C, and from dichloromethane, for example, at about 5°C. In some embodiments, Form I of Formula I has peaks at about 6.1, 8.6 and 16.4°²θ as measured by XRPD. For example, Form I of Formula I can have peaks at about 6.1, 8.6, 12.3, 16.4, 17.4 and 18.4°²θ. For example, Form I of Formula I can have peaks at about 6.1, 8.6, 10.7, 12.3, 16.4, 17.4, 18.4, 20.5 and 23.8°²θ. For example, form I of formula I may have peaks at approximately 6.1, 8.6, 10.7, 12.3, 15.1, 16.4, 17.4, 18.4, 20.5, 22.4, 23.8 and 24.7°2θ. For example, form I of formula I may have the peaks listed in Table 1. In some embodiments, form I of formula I has an X-ray diffraction pattern substantially similar to that shown in Figure 5B or 6B. [Table 1] Form II, Formula I

[0076] Form II of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate (Formula I) can be crystallized from many solvents. These solvents include, but are not limited to, acetone, anisole, 1-butanol, cyclohexane, diisopropyl ether, 1,4-dioxane, ethyl acetate, heptane, hexane, isopropyl acetate, methyl ethyl ketone, methyl isobutyl ketone, methyl tetrahydrofuran, tert-butyl methyl ether, tetrahydrofuran, and toluene and mixtures thereof.

[0077] Various batches of morphological polymorphs are described below.

[0078] Batch "GMP-1" was manufactured on a 65-gram scale using Good Manufacturing Practice (GMP) standards. The crystals had a rod-like morphology, as characterized by polarized light microscopy (PLM).

[0079] Batch "GMP-2" was manufactured on a 60-gram scale using Good Manufacturing Practice (GMP) standards. The crystals had a rod-like morphology, as characterized by polarized light microscopy (PLM).

[0080] Batch "Cooling-1" was prepared on an 8-gram scale, as shown in Example 3 below. This batch was prepared by dissolving fumargyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate in 39 mL of a 50:50 mixture of methyl-tert-butyl ether (MTBE) and methanol. The compound was dissolved at 50°C and slowly cooled to 5°C before adding additional MTBE. Three samples were taken from batch Cooling-1 at (i) before the addition of additional MTBE, (ii) after the addition of 30 mL of MTBE, and (iii) after the addition of 60 mL of MTBE. The XRPD spectra of the three samples taken from batch Cooling-1 are shown in Figures 3A, 3B, and 3C, respectively.

[0081] Batch "Antisol-1" was prepared on an 8-gram scale, as shown in Example 4 below. This batch was prepared by dissolving fumargyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate in 39 mL of a 50:50 mixture of methyl-tert-butyl ether (MTBE) and methanol. The compound was dissolved at 50°C, and additional MTBE was added before cooling to 5°C. Three samples were taken from Antisol-1 at (i) before the addition of additional MTBE, (ii) after the addition of 30 mL of MTBE, and (iii) after the addition of 60 mL of MTBE. The XRPD spectra of the three samples taken from batch Antisol-1 are shown in Figures 4A, 4B, and 4C, respectively.

[0082] In one embodiment, the disclosure provides polymorphs of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate characterized by an X-ray powder diffraction pattern using Cu Kα irradiation, which includes peaks at approximately 6.0, 9.0, and 18.0 (e.g., approximately 5.96, 9.00, and 18.02)°2θ.

[0083] In one embodiment, form II further features an X-ray powder diffraction pattern using Cu Kα irradiation, including peaks at approximately 6.0, 9.0, and 18.0°2θ. In another embodiment, form II further features an X-ray powder diffraction pattern using Cu Kα irradiation, including peaks at approximately 6.0, 9.0, 12.3, 12.5, 16.1, and 18.0°2θ. In other embodiments, form II is further characterized by an X-ray powder diffraction pattern using Cu Kα irradiation, including peaks at approximately 5.4, 6.0, 9.0, 12.3, 12.5, 13.8, 16.1, 17.3, 18.0, 20.0, 20.8, and 25.8 °2θ. In other embodiments, form II is further characterized by an X-ray powder diffraction pattern using Cu Kα irradiation, including peaks at approximately 5.4, 6.0, 9.0, 12.3, 12.5, 13.8, 16.1, 17.3, 18.0, 20.0, 20.8, 21.5, 22.0, 24.3, and 25.8 °2θ. In other embodiments, form II of formula I is further characterized by an X-ray powder diffraction pattern including any of the peaks listed in Tables 2-7.

[0084] In one or more embodiments, Embodiment II further features an X-ray powder diffraction pattern substantially similar to that shown in Figure 1A. In one or more embodiments, Embodiment II further features an X-ray powder diffraction pattern substantially similar to that shown in Figure 2A. In one or more embodiments, Embodiment II further features an X-ray powder diffraction pattern substantially similar to that shown in Figures 3A, 3B, or 3C. In one or more embodiments, Embodiment II further features an X-ray powder diffraction pattern substantially similar to that shown in Figures 4A, 4B, or 4C. In one or more embodiments, Embodiment II further features an X-ray powder diffraction pattern substantially similar to that shown in Figure 5A. In one or more embodiments, Embodiment II further features an X-ray powder diffraction pattern substantially similar to that shown in Figure 6A.

[0085] In one or more embodiments, the polymorph of form II is further characterized by the onset of an exothermic event at approximately 178°C and a peak at approximately 188°C when measured by differential scanning calorimetry. Figures 1B and 2B show DSC plots of form II of formula I. In one or more embodiments, the polymorph of form II is further characterized by the onset of an exothermic event at approximately 181°C and a peak at approximately 189°C when measured by thermogravimetric analysis / differential thermal analysis. Figures 1C and 2C show TG / DTA plots of a sample of form II of formula I.

[0086] As shown in the tables and figures of this application, not all values ​​of the peaks (position 2θ) will necessarily match those of different lots of the polymorphs of this application. A person skilled in the art will understand that even different lots of polymorphs of the same form may produce slightly different characterization data, but not obviously different. For example, slight variations in the calibration of the instrument used to perform a given measurement, or slight variations in relative humidity between measurements, may result in data showing slight differences between lots. A person skilled in the art could therefore calibrate his or her instrument and perform repeated measurements to minimize inconsistencies between signals for properly characterizing the polymorphs of this application. However, despite slight differences between batches, the polymorphs of this application are identified and characterized by the characteristic peaks described above (e.g., about 6.0, 9.0 and 18.0°2θ, e.g., ± 0.2°2θ). XPRD [Table 2] [Table 3] [Table 4] [Table 5] [Table 6] [Table 7]

[0087] Table 8 shows the polarization analysis data for lots GMP-1 and GMP-2 of formula I. [Table 8] Single-crystal X-ray diffraction (SXRD)

[0088] The crystal structure of formula I is shown in Figure 8. All non-hydrogen atoms are shown as thermal ellipsoids set at a 30% occupancy level.

[0089] An asymmetric unit containing protonated fumagillol-6-yl N-(trans-4-aminocyclohexyl)carbamate of two complete formula units having only two related benzenesulfonate anions was found. During refinement, large thermal displacement ellipsoids were observed for the carbon atoms C(20B), C(21B), C(22B) and C(23B) of the alkene moiety of molecule “B”.

[0090] Further irregularities were observed for benzenesulfonate “molecule B” centered on sulfur atom S(1B) which also shows a large thermal displacement ellipsoid for the benzyl ring C(24B)<C(29B). A rotational disorder was observed for the sulfonate moiety of this molecule centered on S(1B), with an occupancy of 0.78:0.22 fitting well, and the distances -O(6B) and S(1B)-O(8C) were fixed at 1.48(1), and 1.44(1) for S(1B)-O(7C).

[0091] Approximately 1.5% of the unit cell is occupied by two solvent voids measuring approximately 168 Å3 and 812 Å3, and the electron density was found to be equal to 74 and 137 electrons respectively. Although not indicative of being constrained by theory, these are most likely arising from disordered solvent molecules. Therefore, the electron density within these voids was modeled with a small mole fraction of solvent molecules. It contains 0.27 equivalents of water, 0.12 equivalents of methanol and 0.06 equivalents of MTBE. Of these, the partially occupied methanol and two partially occupied water molecules were refined at the unit cell origin.

[0092] To correct for the extinction effect of the observed data, an extinction correction of 0.00031(3) was applied, all hydrogen atoms were placed at calculated positions, and refinement was carried out using a riding model where 1.2 times Uiso was fixed for all CH, CH2, NH, NH3 groups and 1.5 times Uiso was fixed for all CH3 groups.

[0093] The highest residual Fourier peak is 0.80 e. Å. 3 Approximately 1.1 Å from C(30) 3 This was the result. While not constrained by theory, this appears to be due to further disorder present in the solvent voids occupied by the modeled MTBE molecule centered around O(9). The deepest Fourier hole was -0.41 e. Å, approximately 1.54 Å from H(9AA).

[0094] Polymorph III of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate can be crystallized from ethanol and isopropanol, for example, at about 50°C to about 5°C. In some embodiments, form III of formula I has XRPD peaks at about 8.3, 12.8, and 15.7°2θ. In some embodiments, form III of formula I has XRPD peaks at about 8.3, 12.8, 14.3, 15.7, 18.5, and 21.8°2θ. In some embodiments, form III of formula I has XRPD peaks at about 8.3, 12.8, 14.3, 15.7, 17.6, 18.5, 21.0, 21.8, and 23.5°2θ. In some embodiments, type III of formula I has XRPD peaks at approximately 8.3, 12.8, 14.3, 15.7, 17.6, 18.5, 19.3, 20.3, 21.0, 21.8, 23.5 and 24.1°2θ. In some embodiments, type III of formula I has XRPD peaks at approximately 8.3, 9.0, 12.8, 14.3, 15.7, 17.6, 18.5, 19.3, 20.3, 21.0, 21.8, 22.2, 23.5, 24.1 and 25.0°2θ. In some embodiments, form III has an X-ray diffraction pattern substantially similar to that shown in Figure 5C or 6C. In some embodiments, type III of formula I has XRPD peaks listed in Table 9. [Table 9] Formula I, Form IV

[0095] Polymorph IV of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate can be crystallized from a mixture of isopropanol and water (10%), for example, at about 0°C to about 5°C. In some embodiments, morph IV has XRPD peaks at about 5.1, 7.3 and 14.0°2θ. In some embodiments, morph IV has XRPD peaks at about 5.1, 7.3, 14.0, 18.2, 18.9 and 24.1°2θ. In some embodiments, morph IV has XRPD peaks at about 5.1, 7.3, 14.0, 17.1, 18.2, 18.9, 23.5, 24.1 and 24.7°2θ. In some embodiments, morph IV has an X-ray diffraction pattern substantially similar to that shown in Figure 5D or 6D. In some embodiments, morph IV of formula I has the XRPD peaks listed in Table 10. [Table 10]

[0096] The polymorphs of this application can have high purity. In one or more embodiments, the purity is determined by nuclear magnetic resonance (e.g., 1 It can be measured by 1H NMR. 1 The 1H NMR conditions are shown in the experimental section below. In some embodiments, fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate is used as shown in Figure 7. 1 It can have an H NMR spectrum. Fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate is as follows: 1 It can have an H NMR spectrum. 1H NMR (500 MHz, CDCl3) δ 7.89-7.87 (2H, m, ArH), 7.47-7.45 (3H, m, ArH), 5.45 (1H, br s, CH), 5.21 (1H, t, J = 6.8 Hz, CH), 3.63 (1H, dd, J = 11.3 2.8 Hz, CH), 3.45 (3H, s, OCH3), 3.36-3.30 (1H, br m), 2.97 (1H, d, J = 4.1 Hz), 2.92 (1H, t, J = 11.7 Hz), 2.59 (1H, t, J = 6.1 Hz, CH), 2.55 (1H, d, J = 4.1 Hz), 2.40-2.35 (1H, m), 2.20-2.14 (1H, m), 2.08-1.93 (8H, m), 1.84 (1H, d, J = 12.9), 1.74 (3H, s, =CCH3), 1.67 (3H, s, =CCH3), 1.48-1.38 (2H, m), 1.28-1.24 (1H, m), 1.22 (2H, s), 1.07 (3H, t, J = 12.6 Hz).

[0097] In one or more embodiments, purity can be measured by high-performance liquid chromatography (HPLC) using an evaporative light scattering detector (ELSD) or a charged aerosol detector (CAD). Typical HPLC experimental conditions are shown in the experimental section below. In some embodiments, fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate has a retention time of approximately 8.75 minutes.

[0098] A pair of HPLC plots are shown in Figure 9. As shown in Figures 9 and 11, the following peaks were observed using HPLC analysis, indicating that fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamate benzenesulfonate had a purity of over 99%. [Table 11]

[0099] In one or more embodiments, the salt (i.e., benzenesulfonate of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamate) and the polymorph of this application (e.g., form II) have a purity greater than about 90% (e.g., about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, about 99.5%, about 99.9%, or about 99.99%). In one or more embodiments, the salt (i.e., benzenesulfonate of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamate) and the polymorph of this application (e.g., form II) have a purity greater than about 95%. In one or more embodiments, the salt (i.e., benzenesulfonate of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamate) and the polymorph of this application (e.g., form II) have a purity higher than about 97%. In one or more embodiments, the salt (i.e., benzenesulfonate of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamate) and the polymorph of this application (e.g., form II) have a purity higher than about 98%. In one or more embodiments, the salt (i.e., benzenesulfonate of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamate) and the polymorph of this application (e.g., form II) have a purity higher than about 99.1% (e.g., about 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, or 99.9%).

[0100] In one or more embodiments, the salt (i.e., benzenesulfonate of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamate) and the polymorph of this application (e.g., form II) contain less than 0.5% aminocyclohexyl impurities. In one or more embodiments, the salt (i.e., benzenesulfonate of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamate) and the polymorph of this application (e.g., form II) contain less than 0.4%, less than 0.3%, less than 0.2%, or less than 0.1% aminocyclohexyl impurities. Form II: Fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamate hydroxynaphthoate

[0101] In some embodiments, the present disclosure provides polymorphs of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamate hydroxynaphthoethate. Form A, Formula II

[0102] Form A of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamate hydroxynaphthoethate can be crystallized from many solvents. These solvents include, but are not limited to, acetonitrile, ethyl acetate, methyl ethyl ketone, and tetrahydrofuran.

[0103] In one aspect, the disclosure provides a polymorph (morph A) of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamate hydroxynaphthoethate (formula II) characterized by X-ray powder diffraction (X-ray powder diffraction) patterns at approximately 5.6, 8.9, and 15.4°2θ. In one aspect, the polymorph is characterized by patterns at approximately 5.6, 8.9, 11.5, 12.1, 15.4, and 20.9°2θ. In one aspect, the polymorph is characterized by patterns at approximately 5.6, 8.9, 11.5, 12.1, 15.4, 15.9, 19.7, 20.9, and 23.3°2θ. In one embodiment, the polymorph is characterized by patterns at approximately 5.6, 8.9, 11.5, 12.1, 14.6, 15.4, 15.9, 17.4, 18.1, 19.7, 20.9, and 23.3°2θ. In one embodiment, the polymorph is characterized by an X-ray powder diffraction pattern including the peaks listed in Table 12 or 13.

[0104] In one or more of the above embodiments, Form A of fumagillin-6-yl N-(trans-4-aminocyclohexyl)carbamate hydroxynaphthoate is further characterized by an X-ray diffraction pattern substantially similar to that described in any one of FIGS. 10A to 10E.

[0105] In one or more of the above embodiments, Form A of fumagillin-6-yl N-(trans-4-aminocyclohexyl)carbamate hydroxynaphthoate is further characterized by an endothermic onset at about 182° C. and a peak at about 186° C., measured by DSC. FIG. 11 shows a differential scanning calorimetry curve of a sample of Form A of Formula II.

[0106] In one or more of the above embodiments, Form A of fumagillin-6-yl N-(trans-4-aminocyclohexyl)carbamate hydroxynaphthoate is characterized by an endothermic onset at about 186° C., a peak at about 187° C., and an exotherm at about 191° C. (peak), measured by thermogravimetric analysis / differential thermal analysis. FIG. 12 shows a thermogravimetric analysis / differential thermal analysis plot of a sample of Form A of Formula II. XPRD [Table 12] [Table 13]

[0107] In one or more of the above embodiments, the compound of Formula II can have an 1 H NMR spectrum as shown in FIG. 13. In some embodiments, the compound of Formula II can have the following 1 H NMR spectrum: 1H NMR (500 MHz, CDCl3) δ 8.39 (1H, d, J = 8.2 Hz, ArH), 7.83 (1H, d, J = 8.8 Hz, ArH), 7.76 (1H, d, J = 8.2 Hz, ArH), 7.54-7.46 (2H, td, J = 27.2 7.3 Hz), 7.23 (1H, d, J = 8.5 Hz), 5.45 (1H. br s), 5.19 (1H, t, J = 6.8 Hz, CH), 3.69 (1H, dd, J = 11.7 2.2 Hz, CH), 3.48 (3H, s, OCH3), 3.19 (1H, t, J = 10.9 Hz), 2.91 (1H, d, J = 3.8 Hz), 2.84 (1H, t, J = 6.3 Hz), 2.46 (1H, d, J = 4.1 Hz, CH), 2.42 (1H, t, J = 7.9 Hz), 2.29 (1H, d, J = 9.8 Hz), 2.20-2.03 (7H, m), 1.98-1.91 (2H, m), 1.77 (3H, s, =CCH3), 1.68 (3H, s, =CCH3), 1.52-1.36 (4H, m), 1.20 (3H, s, CCH3), 0.88 (1H, d, J = 13.2 Hz).

[0108] In one or more embodiments, the compound of formula II may have HPLC plots as shown in Figures 14A and 14B. As shown in Figure 14A, the upper plot is a low-resolution plot of the mother liquor, and the lower plot shows the same analysis at a higher resolution. The peaks corresponding to Figure 14A are shown in Table 14A below. As shown in Figure 14B, the upper plot is a low-resolution plot of the solid formula II product, and the lower plot shows the same analysis at a higher resolution. The peaks corresponding to Figure 14B are shown in Table 14B below. [Table 14] [Table 15]

[0109] In one or more embodiments, the salts presented herein (i.e., benzenesulfonate and hydroxynaphthoate of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamate) and the polymorphs of this application (e.g., Form II and Form A) contain less than 0.5% of the following compound 2: [ka] In one or more embodiments, the salt (i.e., benzenesulfonate of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamate) and the polymorph of this application (e.g., form II) contain less than 0.4%, less than 0.3%, less than 0.2%, or less than 0.1% of compound 2.

[0110] In one or more embodiments, the salt (i.e., benzenesulfonate of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamate) and the polymorphs of this application (e.g., Form II and Form A) contain less than 0.5% of compound 3. [ka] In one or more embodiments, the salt (i.e., benzenesulfonate of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamate) and the polymorph of this application (e.g., form II) contain less than 0.4%, less than 0.3%, less than 0.2%, or less than 0.1% of compound 3.

[0111] In one or more embodiments, the salt (i.e., benzenesulfonate of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamate) and the polymorphs of this application (e.g., Form II and Form A) contain less than 0.5% of compound 4. [ka] In one or more embodiments, the salt (i.e., benzenesulfonate of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamate) and the polymorph of this application (e.g., form II) contain less than 0.4%, less than 0.3%, less than 0.2%, or less than 0.1% of compound 4.

[0112] In one or more embodiments, the salt (i.e., benzenesulfonate of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamate) and the polymorphs of this application (e.g., Form II and Form A) contain less than 0.5% of compound 5. [ka] In one or more embodiments, the salt (i.e., benzenesulfonate of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamate) and the polymorph of this application (e.g., form II) contain less than 0.4%, less than 0.3%, less than 0.2%, or less than 0.1% of compound 5.

[0113] In one or more embodiments, the salt (i.e., benzenesulfonate of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamate) and the polymorphs of this application (e.g., Form II and Form A) contain less than 0.5% of compound 6. [ka] In one or more embodiments, the salt (i.e., benzenesulfonate of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamate) and the polymorph of this application (e.g., form II) contain less than 0.4%, less than 0.3%, less than 0.2%, or less than 0.1% of compound 6.

[0114] In one or more embodiments, the salt (i.e., benzenesulfonate of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamate) and the polymorphs of this application (e.g., Form II and Form A) are highly stable. Stability as defined herein is understood to mean that the compound is reliably maintained at a high level of purity over a long period of time. In some embodiments, stability can be defined as remaining at a purity of more than 95% for at least six months. In some embodiments, the polymorphs of this application can maintain their purity for long periods of time, and even at high temperatures and / or relative humidity. In some embodiments, the polymorphs of this application can maintain their purity for more than one month, more than two months, more than three months, or more than six months, and even at high temperatures (e.g., more than 25°C, more than 30°C, more than 35°C) and / or relative humidity (e.g., more than 50% RH, more than 60% RH, more than 75% RH, or more than 90% RH). In some embodiments, the compounds disclosed (e.g., form II of formula I and form A of formula II) also retain their purity over long periods even at low temperatures. For example, the compounds disclosed can retain their purity for 6 months or longer at -20°C, or for 6 months or longer at about 0-5°C.

[0115] In some embodiments, the polymorphs of this application can maintain their purity for at least one month; at least two months; at least three months; at least four months; at least five months; at least six months; at least one year; and at least two years. In some embodiments, the polymorphs of this application can maintain their purity at about 25 °C; about 30 °C; about 35 °C; about 40 °C; about 45 °C; and about 50 °C. In some embodiments, the polymorphs of this application can maintain their purity at about 60% relative humidity; about 65% relative humidity; about 70% relative humidity; about 75% relative humidity; about 80% relative humidity; about 85% relative humidity; about 90% relative humidity; about 90% relative humidity; about 95% relative humidity; and about 100% relative humidity.

[0116] In some embodiments, the compounds of the present disclosure are stable at approximately 5 °C ± 3 °C, approximately 25 °C ± 2 °C and 60% relative humidity ± 5% relative humidity, approximately 30 °C ± 2 °C and 65% relative humidity ± 5% relative humidity, and approximately 40 °C ± 2 °C and 75% relative humidity ± 5% relative humidity.

[0117] Table 15 below shows the purity data for Form II of Formula I after storage at 25°C and 60% relative humidity for 6 months, and Table 16 below shows the purity data for Form II of Formula I after storage at 40°C and 75% relative humidity for 6 months. As shown below, the compound can maintain a purity of over 99% after being stored at 25°C and 60% relative humidity for 6 months. The compound can maintain a purity of over 95% after being stored at 40°C and 75% relative humidity for 6 months. Table 15. Stability of Form II of Compound I at 25 °C and 60% relative humidity (RH). [Table 16] Table 16. Stability of Form II of Compound I at 40°C and 75% relative humidity (RH) [Table 17] Method for preparing polymorphs

[0118] Polymorph II of Formula I can be prepared by recrystallizing fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate from a number of different solvents. In some preferred embodiments, the polymorph of form II is prepared from a mixture of MTBE and / or methanol. In some embodiments, the ratio of MTBE to methanol is 50:50. Two methods for producing the polymorph of form II by recrystallizing fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate are described below.

[0119] In any of the embodiments described below, the methanol to methyl tert-butyl ether ratio can be any ratio between 1:99 and 99:1. In any of the embodiments described below, step 1 can be carried out at any temperature (e.g., between about 25°C and about 60°C; or between about 40°C and about 60°C). In any of the embodiments described below, step 1A may include cooling the solution to about 25°C (e.g., a temperature of about 40°C). In some embodiments, step 1A may include cooling the solution between about 0°C and 10°C (e.g., between about 2°C and about 8°C). In some embodiments, step 2A may include further cooling of the solution. For example, step 1A may cool the solution to about 25°C, and step 2A may further cool the solution to about 2°C and about 8°C. In some embodiments, step 1A may be omitted, and step 2A may include cooling the solution between about 40°C and about 25°C or between about 2°C and 8°C.

[0120] In some embodiments, this disclosure is as follows: (i) Step 1: Dissolve fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate in methanol, optionally containing MTBE, to form a solution; (ii) Step 2: If applicable, add a seed crystal of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate (e.g., about 1% by weight) to the solution; (iii) Step 3: If applicable, separate the crystallized fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate from the solution. The present invention provides a method for preparing a polymorph (form II) of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate (form I), including the steps involved.

[0121] In some embodiments, the method may be performed after step 1 but before step 2, with steps 1A and / or 1B: Step 1A: Cool the fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate solution obtained from the previous step; Step 1B: Add additional MTBE to the fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate solution obtained from the previous step; It also includes.

[0122] In some embodiments, the method may be performed after step 2 but before step 3, with steps 2A and / or 2B: Step 2A: Cool the fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate solution obtained from the previous step; Step 2B: Add MTBE to the fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate solution obtained from the previous step; It also includes.

[0123] In one embodiment, step 1 is carried out at ambient temperature or room temperature (e.g., about 20°C to about 25°C). In one embodiment, step 1 includes heating the fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate in methanol, optionally containing MTBE (e.g., to about 38 to 50°C or about 40°C) to promote dissolution.

[0124] Polymorphs of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate (formula I) can be prepared by carrying out steps 1, 2 and 3 and any combination of steps 1A, 1B, 2A and 2B as described above.

[0125] In one embodiment, this disclosure is as follows: (1) Add fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate to a certain amount of methanol to form a solution; (1B) Add a certain amount of tert-butylmethyl ether to the solution; (1A) Cool the solution; (2) Add a seed crystal of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate (for example, about 1% by weight) to the solution; (2A) Cool the solution; (2B) Add an additional tert-butyl methyl ether to the solution to produce a crystalline form of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate; and (3) Filter the solution to isolate the crystalline fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate. The present invention provides a method for preparing polymorphs of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate (e.g., form II), including the above.

[0126] In one or more embodiments of the above configuration, the ratio of methanol to fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamate benzenesulfonate in step (1) is approximately 5:3 (liters / mol). In one or more embodiments, approximately 1 liter of methanol is used to dissolve approximately 0.6 moles of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamate benzenesulfonate in step (i). In one or more embodiments of the above method, the mixture in step (1) is heated to approximately 38-50°C to form a solution. In one or more embodiments of the above method, the mixture in step (1) is heated to, for example, approximately 40°C to form a solution.

[0127] In one or more embodiments, the volume of tert-butyl methyl ether added in step (1B) is approximately 1 to 1.5 times the volume of methanol used in step (1). In one or more embodiments, the volume of tert-butyl methyl ether added in step (1B) is approximately 1.25 times the volume of methanol used in step (1). In one or more embodiments, the solution becomes turbid (cloudy) after the addition of tert-butyl methyl ether.

[0128] In one or more embodiments, step (1A) includes cooling the solution to about 30-36°C. In one or more embodiments, step (1A) includes cooling the solution to about 35°C. In one or more embodiments, step (1A) is performed slowly. In one or more embodiments, the cooling is completed in about 1 hour.

[0129] In one or more embodiments, the seed crystal of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate (e.g., about 1% by weight) added in step (2) is pulverized before being added.

[0130] In one or more embodiments, step (2A) includes cooling the solution to about 2–8°C. In one or more embodiments, step (2A) further includes maintaining the temperature of the solution (e.g., at about 2–8°C) for a certain period of time (e.g., about 5–20 minutes). In one or more embodiments, step (2A) further includes maintaining the temperature of the solution (e.g., at about 2–8°C) for about 10 minutes.

[0131] In one or more embodiments, the volume of additional tert-butyl methyl ether added in step (2B) is about 4 to 8 times the volume of methanol used in step (1). In one or more embodiments, the volume of additional tert-butyl methyl ether added in step (2B) is about 6.5 times the volume of methanol used in step (1). In one or more embodiments, the temperature in step (2B) is maintained at about 2 to 8°C. In one or more embodiments, the temperature in step (2B) is maintained at about 2 to 8°C for a certain period of time (e.g., about 1 hour).

[0132] In one or more embodiments, the crystalline form of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate is further washed with tert-butyl methyl ether after step (3). In one or more embodiments, the crystalline form of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate can be further dried under vacuum. In one or more embodiments, the crystalline form of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate can be further dried under vacuum at about 40°C.

[0133] In one or more embodiments, the steps provided herein, or combinations thereof (e.g., step 1, step 1', step 1A, step 1B, step 2, step 2A, step 2B, step 3) for producing Form II and Form A can result in increased purity and / or increased yield. In some embodiments, the application of the steps provided herein can improve the yield compared to simply applying steps 1, 2 and 3. For example, the application of the steps herein can improve the yield by about 10%, about 20%, about 30%, 40%, about 50%, about 60%, about 70%, about 80%, about 90%, or about 100%. The yield of Form II or Form A can be about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98%, about 99%, about 99.5%, or about 99.9% or higher. Furthermore, the purity of Form II or Form A can be improved by applying the methods provided herein (e.g., Step 1, Step 1', Step 1A, Step 1B, Step 2, Step 2A, Step 2B, Step 3). In some embodiments, the application of the steps provided herein can improve purity more than simply applying Steps 1, 2, and 3. For example, the application of the steps herein can improve purity by about 10%, about 20%, about 30%, 40%, about 50%, about 60%, about 70%, about 80%, about 90%, or about 100%. The purity of Form II or Form A is about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98%, about 99%, about 99.5%, or about 99.9% or higher.

[0134] In one embodiment, this disclosure is as follows: (1) Dissolve fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate in a mixture of tert-butyl methyl ether and methanol to form a solution; (1A) Cool the solution; (2) Add a seed crystal of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate (for example, about 1% by weight) to the solution; A certain amount of tert-butyl methyl ether is added to the solution; (2A) Continue to cool the solution; (2B) Add an additional amount of tert-butyl methyl ether to the solution in order to produce the crystalline form of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate; and (3) Filter the solution to isolate the crystalline form of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate. The present invention provides a method for preparing polymorphs of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate (e.g., form II), including the above.

[0135] In one or more embodiments, the ratio of tert-butyl methyl ether to methanol in step (1) is between approximately 50:50 and approximately 20:80. In one or more embodiments, the ratio is approximately 50:50. In one or more embodiments, dissolution is carried out at approximately 40 to 60°C. In one or more embodiments, dissolution is carried out at approximately 50°C. In one or more embodiments, the solution of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate prepared in step (i) is stirred for approximately 15 minutes to 1 hour. In one or more embodiments, the solution of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate prepared in step (1) is stirred at 50°C for approximately 30 minutes to ensure complete dissolution before cooling.

[0136] In one or more embodiments, step (1A) includes cooling at a rate of approximately 0.1 to 0.5°C / min. In one or more embodiments, step (1A) includes cooling at a rate of approximately 0.2°C / min. In one or more embodiments, step (ii) includes cooling to approximately 30 to 45°C. In one or more embodiments, step (1A) includes cooling to approximately 37°C.

[0137] In one or more embodiments, the seed crystal of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate (e.g., about 1% by weight) added in step (2) is pulverized before being added.

[0138] In one or more embodiments, step (2A) includes cooling at a rate of approximately 0.1 to 0.5°C / min. In one or more embodiments, step (2A) includes cooling at a rate of approximately 0.2°C / min. In one or more embodiments, step (2A) includes a step of cooling to less than 10°C. In one or more embodiments, step (2A) includes cooling to approximately 5°C.

[0139] In one or more embodiments, the amount of tert-butyl methyl ether added in step (2B) is about 10-50% of the amount of the tert-butyl methyl ether and methanol mixture used in step (1). In one or more embodiments, the amount of tert-butyl methyl ether added in step (2B) is about 15% of the amount of the tert-butyl methyl ether and methanol mixture used in step (1). In one or more embodiments, the amount of tert-butyl methyl ether added in step (2B) is about 30% of the amount of the tert-butyl methyl ether and methanol mixture used in step (1).

[0140] In one or more embodiments, the crystalline form of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate is further washed with tert-butyl methyl ether after step (3). In one or more embodiments, the crystalline form of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate can be further dried under vacuum. In one or more embodiments, the crystalline form of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate can be further dried under vacuum at about 40°C.

[0141] In one or more embodiments, this disclosure is as follows: (1) Dissolve fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate in a mixture of tert-butyl methyl ether and methanol to form a solution; (1B) Add tert-butylmethyl ether to the solution; (2) Add a seed crystal of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate (for example, about 1% by weight) to the solution; (2B) Add additional tert-butylmethyl ether to the solution; (2A) Cool the solution to produce a crystalline form of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate; and (3) Filter the solution to isolate the crystalline form of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate. The present invention discloses a method for preparing a polymorph (e.g., form II) of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate, including the above.

[0142] In one or more embodiments, the ratio of tert-butyl methyl ether to methanol in step (1) is between approximately 50:50 and approximately 20:80. In one or more embodiments, the ratio is approximately 50:50. In one or more embodiments, the dissolution is carried out at approximately 40 to 60°C. In one or more embodiments, the dissolution is carried out at approximately 50°C. In one or more embodiments, the solution of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate produced in step (1) is stirred for approximately 15 minutes to 1 hour. In one or more embodiments, the solution of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate produced in step (1) is stirred at 50°C for approximately 30 minutes to ensure complete dissolution before cooling.

[0143] In one or more embodiments, the volume of tert-butyl methyl ether added in step (1B) is approximately 1 to 1.5 times the volume of methanol used in step (1). In one or more embodiments, the volume of tert-butyl methyl ether added in step (1B) is approximately 1.25 times the volume of methanol used in step (1). In one or more embodiments, the solution becomes turbid (cloudy) after the addition of tert-butyl methyl ether.

[0144] In one or more embodiments, the seed crystal of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate (e.g., about 1% by weight) added in step (2) is pulverized before being added.

[0145] In one or more embodiments, the amount of tert-butyl methyl ether added in step (2B) is about 10-50% of the amount of the tert-butyl methyl ether and methanol mixture used in step (1). In one or more embodiments, the amount of tert-butyl methyl ether added in step (2B) is about 15% of the amount of the tert-butyl methyl ether and methanol mixture used in step (1). In one or more embodiments, the amount of tert-butyl methyl ether added in step (2B) is about 30% of the amount of the tert-butyl methyl ether and methanol mixture used in step (1).

[0146] In one or more embodiments, step (2A) includes cooling at a rate of approximately 0.1 to 0.5°C / min. In one or more embodiments, step (2A) includes cooling at a rate of approximately 0.2°C / min. In one or more embodiments, step (2A) includes a step of cooling to less than 10°C. In one or more embodiments, step (2A) includes cooling to approximately 5°C.

[0147] In one or more embodiments, the crystalline form of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate is further washed with tert-butyl methyl ether after step (3). In one or more embodiments, the crystalline form of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate can be further dried under vacuum. In one or more embodiments, the crystalline form of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate can be further dried under vacuum at about 40°C. cooling recrystallization

[0148] In some embodiments, polymorph II of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate can be crystallized from a mixture of MTBE and methanol by mixing (trans-4-aminocyclohexyl)carbamatebenzenesulfonate in MTBE and methanol at about 50°C, cooling the resulting mixture, and then adding MTBE to the mixture to complete crystallization. In some embodiments, the mixture of MTBE and methanol is a 50:50 mixture. The ratio of MTBE to methanol can be any ratio between 1:99 and 99:1. In some preferred embodiments, the ratio of MBTE to methanol is less than about 8:2. In some embodiments, the ratio of MBTE to methanol is about 8:2 to about 9:1. In some embodiments, the purity of the resulting fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate may be adversely affected if significantly larger amounts of MBTE are used. In some embodiments, the purity of the resulting fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate may be adversely affected if the ratio of MBTE to methanol is greater than about 9:1. In some embodiments, fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate is dissolved at a temperature of about 50°C. Fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate can also be dissolved at any specific temperature between about 40°C, about 35°C, about 25°C, or 50°C–5°C. The XRPD spectra of the material produced by the cooling recrystallization procedure are shown in Figure 3. Figure 3A shows the XRPD spectrum of the material from the first sample (i.e., before the addition of further MTBE), and Figure 3B shows the material from the second sample (i.e., after the addition of 30 mL of MTBE). Figure 3C shows the XRPD spectrum of the dried final material. Recrystallization using a poor solvent

[0149] In some embodiments, polymorph II of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate can be crystallized from a mixture of MTBE and methanol by adding solid fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate to MTBE and methanol at about 50°C, and adding additional MTBE before cooling the resulting mixture to complete crystallization. The ratio of MTBE to methanol can be any ratio between 1:99 and 99:1. In some embodiments, fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate is dissolved at a temperature of about 50°C. Fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate can also be dissolved at any specific temperature between 50°C and 40°C, such as about 40°C, about 35°C, about 25°C, or 50°C. Figure 4A shows the XRPD spectrum of the substance from the first sample (i.e., after the addition of approximately 30 mL of MTBE), and Figure 4B shows the XRPD spectrum of the substance from the second sample (i.e., after the addition of 60 mL of MTBE). Figure 4C shows the XRPD spectrum of the dried final substance. Crystallization from ethyl acetate

[0150] In some embodiments, formulas I and / or II can be crystallized directly from ethyl acetate (for example, as described in Example 5 below). In some embodiments, the free bases of formulas I and / or II can be crystallized directly from ethyl acetate without acidifying the aqueous layer and extracting the free bases back into the organic solvent, before basicizing the aqueous layer and extracting the free bases back into the organic solvent. This has the advantage of being easier to operate than recrystallization from methanol and / or in some cases. It may also have the advantage of using less organic solvent while still maintaining the purity of the final product (i.e., formulas I and / or II).

[0151] As a result, in some embodiments, this disclosure is as follows: (i) Dissolve fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamate in ethyl acetate to form a solution; (ii) Add benzenesulfonic acid to the ethyl acetate solution of the fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamate; and, (iii) The obtained fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate is filtered. The present invention provides a method for preparing polymorphs (e.g., form II) of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate, including the above.

[0152] In some embodiments, benzenesulfonic acid is added as a solution in ethyl acetate. In some embodiments, benzenesulfonic acid is added at a temperature of about 8–12°C. In some embodiments, this method further includes cooling the mixture of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamate and benzenesulfonic acid to about 0–5°C. Thus, in some embodiments, crystallization from ethyl acetate is reactive crystallization. In some embodiments, the water content in the ethyl acetate free base solution before adding the benzenesulfonic acid / ethyl acetate solution is in the range of 1.0–1.3%.

[0153] Synthesis of Fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate (Formula I) As shown in Scheme 1 below, fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamate benzenesulfonate can be produced by treating fumagyl-6-yl, p-nitrophenyl carbonate with an excess amount of trans-1,4-diaminocyclohexane. The free base product is finally extracted in MTBE and treated with benzenesulfonic acid as shown in Example 1 below. Scheme 1. Synthesis of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate from fumagyl-6-yl, p-nitrophenyl carbonate. [ka]

[0154] While not intended to imply theoretical constraints, the free base form of formula I, i.e., fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamate, is inherently unstable due to the presence of both the primary amine and the two epoxides. Furthermore, the hemi-tartrate of formula I (not shown) was found to contain significant amounts of the autocondensation products, compounds 2 and 3, as well as the hydrolysis product, compound 4, as shown above. The hemi-tartrate and benzenesulfonate also contained impurities associated with compound 5 (shown above), which is the starting material for fumagiline (a natural product). Extensive experimental studies have not been able to reduce the impurity levels in the hemi-tartrate to pharmaceutically acceptable levels. Therefore, the fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamate benzenesulfonate and its polymorphs (e.g., form II) of this disclosure are understood to be more stable than the corresponding hemi-tartrate and free base.

[0155] In contrast to the free base and other salt forms described above, the polymorphs described herein (e.g., form II) are stable at standard temperatures and pressures established, for example, by ICH standards for the evaluation of pharmaceutical substances. Therefore, the polymorphs described herein can be used as stable chemical intermediates, as shown below.

[0156] In some embodiments, the present disclosure provides the reactive crystallization of compounds of formula I from the corresponding free bases described in Example 1 and / or Example 5. This method avoids the isolation of free base epoxides that undergo autocondensation in some embodiments. Use of Fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate

[0157] Fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate (Formula I) is useful as an intermediate in the synthesis of polymer conjugate derivatives of fumagilol or fumagiline, such as the compound of Formula III: [ka]

[0158] In one or more embodiments, this disclosure relates to compound 1 [ka] A method for producing a compound represented by the following formula (I) or formula (II): [ka] [ka] The compound represented by the following formula (IV): [ka] (wherein x is an integer between 1 and about 450; y is an integer between 1 and about 30; n is an integer between 1 and about 50; and LG is a leaving group, and in one embodiment, the leaving group LG is, [ka] (is) The present invention provides a method that includes bringing into contact with [something].

[0159] Compound III can be prepared from compound I according to Scheme 2 below. Formula I is treated with HPMA copolymer (which itself is bonded to an amino acid chain) as shown in Scheme 2 below. The amino acid chain has a suitable leaving group (LG, e.g., para-nitrophenyl), and the reaction is carried out in the presence of a suitable solvent (e.g., DMF). Scheme 2. Example of preparation of compound I from formulas I to III. [ka]

[0160] Certain impurities during the synthesis of compounds of formula I or II, such as compounds 2, 3, 4, and 5 mentioned above, can also be reacted in the same manner as those of formula I or II and bound to the polymer. This can interfere with the reaction and reduce the purity of formula III because polymer-bound impurities are difficult to remove. The polymorphs of this application, as described above, have high purity and contain particularly small amounts of compounds 2, 3, 4, and 5. Therefore, the polymorphs of this application are particularly suitable as intermediates for the preparation of the compounds and can be used to produce formula III in good yield and high purity. Not to imply being bound by theory, the use of salts presented herein (e.g., formula I or formula II) as intermediates for compounds of formula III may provide higher yield and higher purity compared to the use of the corresponding free bases.

[0161] Compounds of formula I or formula II and the polymorphs of this application may be useful for inhibiting MetAP2. In some embodiments, compounds of formula I or formula II or the polymorphs of this application can be used as MetAP2 inhibitors. Thus, compounds of formula I or formula II or the polymorphs of this application (e.g., morph II, morph A) can be used to treat diseases or conditions in which MetAP2 is involved (e.g., onset, regulation, and / or progression of disease or symptoms), such as cancer (e.g., hematological cancers such as lymphoma). For example, Figure 15 shows the effects of the compounds of this disclosure on cancer cells. Figure 15A shows the effect of formula I on B-cell lymphoma cells as a concentration function. Figure 15B shows the effect of formula I on mantle cell lymphoma cells as a concentration function. Figure 15C shows the effect of formula I on diffuse large B-cell lymphoma cells as a concentration function. Figure 15D shows the effect of formula I on myeloma cells as a concentration function.

[0162] Another aspect of this disclosure relates to a pharmaceutical composition comprising a compound of formula I and / or formula II and a pharmaceutically acceptable carrier or excipient.

[0163] Another aspect of the present disclosure relates to a pharmaceutical composition comprising a polymorph of the present application (e.g., polymorphic form II of formula I) and a pharmaceutically acceptable carrier.

[0164] When providing one or more compounds or polymorphs of this application to a mammal, the dosage of the compound or polymorph administered depends on the mammal's age, weight, height, sex, general medical condition, prior medical history, disease progression, tumor burden, route of administration, prescription, etc. For example, an appropriate dose of the compounds of this disclosure for a mammal requiring the treatment described herein ranges from about 0.01 mg to about 2000 mg of the compound per kilogram of body weight. Furthermore, in the case of Formula II, due to the effect of binding to the polymer, the drug may be administered at a lower dose than typically used in the treatment of a particular disorder. Surprisingly, in some embodiments, the polymer conjugates of this disclosure are more active on a weight / weight basis than the corresponding low molecules.

[0165] This disclosure is further illustrated by the following examples, but should not be construed as limiting the scope or idea of ​​the specific procedures described herein. It should be understood that these examples are provided to illustrate specific embodiments and do not limit the scope of this disclosure. It should be further understood that various other embodiments, modifications, and equivalents that can be suggested to those skilled in the art without departing from the intent of this disclosure and / or the appended claims may be given to the means. All solvents and other chemical reagents were obtained from commercial sources (e.g., Sigma-Aldrich (St. Louis, MO)) and used without further purification unless otherwise stated. Abbreviation

[0166] [Table 18] XRPD analysis

[0167] Unless otherwise specified, XRPD analysis was performed using a panalytical X'pert pro, scanning the sample between 3 and 35°²θ. The material was gently ground and supported on a multiwell plate with a Mylar polymer film. The multiwell plate was then loaded into a diffractometer operating in transmission mode and analyzed under the following experimental conditions: Pre-processing raw data: XRD measurement (*.XRDML) Scan Axis: Goniometer Start Position [°2θ]: 3.0066 End Position [°2θ]: 34.9866 Step size [°2θ]: 0.0130 Scan step time [s]: 18.8700 Scan type: Continuous PSD Mode: Scanning PSD length [°2θ]: 3.35 Offset [°2θ]: 0.0000 Divergence slit type: Fixed Divergence slit size [°]: 1.0000 Measurement temperature [°C]: 25.00 Anode material: Cu K-α1[Å]: 1.54060 K-α2[Å]: 1.54443 K-β[Å]: 1.39225 K-A2 / K-A1 ratio: 0.50000 Generator Setting: 40 mA, 40 kV Goniometer radius [mm]: 240.00 Illumination distance - Divergence slit [mm]: 91.00 Incident Beam Monochromator: No Spinning: No Thermogravimetric analysis and differential thermal analysis (TGA / DTA)

[0168] Unless otherwise specified, TG / DTA was performed according to the following procedure. Approximately 3 mg of the material was weighed into an open aluminum pan and placed in a thermogravimetric / differential thermal analyzer (TG / DTA), where it was kept at room temperature. The sample was then heated from 25°C to 300°C at a rate of 10°C / min, during which the change in sample weight was recorded along with any differential thermal events (DTA). 300 cm³ was used as the packing gas. 3 Nitrogen was used at a flow rate of [number] minutes. Differential Scanning Calorimetry (DSC)

[0169] Unless otherwise specified, DSC was performed using the following procedure. Approximately 2 mg of material was weighed into an aluminum DSC pan and sealed airtight with a perforated aluminum lid. The sample pan was then cooled and loaded into a Seiko DSC6200 (equipped with a cooler) maintained at 20°C. Once a stable heat flow response was obtained, the sample and reference material were heated to 220°C at a rate of 10°C / min, and the resulting heat flow response was monitored. The sample was cooled back to 20°C, reheated to 220°C, and the resulting heat flow response was monitored. Polarized light microscopy (PLM)

[0170] Approximately 1 mg of the solid was added to a glass microscope slide. The solid was suspended in a drop of silicone oil and covered with a glass coverslide. The sample was analyzed at 20x magnification using both polarized and unpolarized light. polarimetry

[0171] A 10 mg / mL methanol solution was prepared for each sample. 100 μL aliquots were taken from each solution and used to prepare a 1 mg / mL methanol solution. The diluted solutions were used to confirm the results of the tests performed on the 10 mg / mL solution.

[0172] The analysis was performed using a Schmidt and Haensch Polartronic H532 polarimeter. Samples were analyzed using a 100 mm cell. The samples were analyzed using light at a wavelength of 589 nm and a temperature of 26 °C. Specific rotation was calculated using the following formula.

[0173] αD26=(measurement angle x 100) / (concentration x transmission length) X-ray single crystal analysis (SXRD)

[0174] SXRD analysis was performed using a monochromatic MoKα (λ=0.71073Å) diffractometer with an Agilent Technologies (Dual Source) SuperNova diffractometer. The diffractometer was equipped with an Oxford Cryosystems cryogenic device, allowing data acquisition at 120(10)K. The acquired data were corrected for absorption effects based on a Gaussian integration software package using a polyhedral crystal model implemented as part of CrysAlisPro (Agilent Technologies, 2014).

[0175] The aforementioned structure was analyzed by the direct method (SHELXS97)1 and unfolded by full least-squares refinement using F2(SHELXL97)1, interfaced via the OLEX2 software package. Images created for this report were produced via a local program.

[0176] Data were collected, elucidated, and refined in the trigonal space group R3. Searching for higher symmetries using the ADDSYMM2 routine in PLATON3 did not reveal any higher symmetries.

[0177] All fully occupied non-hydrogen atoms were placed within the Fourier map, and their positions were refined before being refined as an anisotropic ellipsoid to describe their thermal behavior. 1 H nuclear magnetic resonance ( 1 (H NMR)

[0178] ¹H NMR spectra were recorded using a Bruker PRO500 (500 MHz) or Bruker AVA500 (500 MHz) spectrometer. Chemical shifts (δ) are cited in parts per million (ppm) of tetramethylsilane under a downfield, with residual protonated solvent as an internal standard (7.26 ppm CDCl3). Experiments were conducted in deuterated chloroform, with each sample prepared at a concentration of approximately 10 mM. High-performance liquid chromatography (HPLC)

[0179] Unless otherwise specified, HPLC was performed under the following experimental conditions: Column: Agilent Eclipse XDB C18 5μm, 150x4.6 mm column Mobile phase A: 0.1% Trifluoroacetic Acid Mobile phase B: 0.1% Trifluoroacetic Acid in Acetonitrile Diluent 1:1 water:acetonitrile Flow rate: 1.0 mL / min Runtime: 24 minutes Detector: Evaporative Light Scattering Detector (ELSD) Detection pressure: 3.6 bar Detection gain: 8 Detection temperature: 50 °C Detection gas: nitrogen Column temperature: 25°C Autosampler temperature: 5 °C Injection volume: 10 μL (sample volume may be adjusted as needed) Concentration gradient program:

Table 19

[0180] Synthesis and Reaction Crystallization of Fumagillin-6-yl N-(trans-4-aminocyclohexyl) Carbamate Benzenesulfonate in Example 1

Chemical formula

[0181] A solution of trans-1,4-diaminocyclohexane (102.2 g, 895 mmol, 4.9 equivalents) dissolved in ethyl acetate (2.8 L) was cooled to 0-5°C. Fumagyl-6-yl p-nitrophenyl carbonate (80.8 g, 181 mmol, 1 equivalent) was dissolved in ethyl acetate (3.2 L) and added to the stirred trans-1,4-diaminocyclohexane solution over 30-90 minutes while maintaining the temperature at 0-5°C. Under in-process control by HPLC-UV, the reaction mixture was stirred at 0-5°C for at least 120 minutes until the carbonate concentration was less than 2% of the starting concentration. The batch was filtered to remove solid by-products, and the solid was washed with ethyl acetate (2 × 1.2 L). The ethyl acetate solution was washed with water (0.5 L), and the aqueous phase was discarded. Ethyl acetate was extracted with a solution of water (1.6 L) and 10% citric acid (0.24 L). The pH of the aqueous phase was approximately 6, and the ethyl acetate was discarded. The aqueous solution was adjusted to pH 10.3 using 1N sodium carbonate (0.87 L). The product in the form of a free base was extracted into MTBE (4 × 1.6 L). The organic phase was dried with sodium sulfate (4 × 128 g) as a drying agent. The organic extracts were combined and filtered to remove the drying agent. The drying agent was washed with MTBE (1.4 L). Fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamate (free base) in MTBE was cooled to 0-5°C, and fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamate benzenesulfonate (0.8 g) was added as a seed. While maintaining the batch at 0-5°C, a solution of benzenesulfonic acid (17.7 g, 0.62 equivalents) in MTBE (0.43 L) was slowly added. The batch was cooled to -20°C. The solid was collected by filtration, washed with MTBE (0.5 L), and dried under reduced pressure at 35°C or below to obtain fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate (68.3 g, 62%, formula I). Example 2: Recrystallization of Fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate

[0182] A stirred suspension of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate (68.3 g, 118 mmole, Formula I) in methanol (200 mL) was heated at 40°C until a clear solution was obtained. While maintaining the batch temperature at 40°C, MTBE (250 mL) was slowly added until the solution became cloudy. The batch was cooled to 35°C over 1 hour. A seed of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate (684 mg, 1.2 mmol, 1 wt%) was added. The batch was cooled in 5°C steps, with 30 minutes allowed at each step and held at 2–8°C for 10 minutes until the batch reached 2–8°C. Additional MTBE (total 8.8 volume) was added over 60 minutes, and the batch was held at 2–8°C for at least 60 minutes. The solid was collected by filtration, washed with MTBE (410 mL), and dried under reduced pressure at 35°C or below to obtain the desired product (64.6 g, 95%) as a polymorph of form II. Example 3. Cooling crystallization of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate

[0183] Fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate (8.1992 g) was dissolved in 39 mL of MTBE:methanol (50:50) to obtain an initial concentration of approximately 210 mg / mL. The solution was stirred at 50°C for 30 minutes and monitored by focused beam reflectance measurement (FBRM) to ensure dissolution. The solution was cooled to 5°C at a rate of 0.2°C / min. At 37°C, 0.08 g (approximately 1%) of the seed material was gently ground using a mortar and pestle and added to the mixture. A sample was taken when the solution temperature reached 5°C, and MTBE was added at a rate of 30 mL / hour until 30 mL of MTBE had been added. A second sample was taken, and the addition of MTBE was continued until 60 mL had been added. Crystallization was then stirred for a further 15 minutes. The final slurry was filtered, and the obtained material was vacuum-dried at 40°C for approximately 16 hours.

[0184] Figure 3 shows the XRPD spectra of the substances produced by the above procedure. Figure 3A shows the XRPD spectrum of the substance from the first sample (i.e., before the addition of further MTBE), and Figure 3B shows the XRPD spectrum of the substance from the second sample (i.e., after the addition of 30 mL of MTBE). Figure 3C shows the XRPD spectrum of the dried final product. Example 4: Antisolvent crystallization of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate

[0185] Fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate (8.0514 g) was dissolved in 39 mL of MTBE:methanol (50:50) to obtain an initial concentration of approximately 210 mg / mL. The solution was stirred at 50°C for 30 minutes and monitored by focused beam reflectance measurement (FBRM) to ensure dissolution. The MTBE was added to the container at a rate of 30 mL / hour. After adding 10.4 mL of MTBE, 0.08 g (approximately 1%) of the seed material was gently ground in a mortar and pestle and added to the mixture. A sample was taken after adding 30 mL of MTBE. The addition of MTBE was continued, and a second sample was taken after adding 60 mL of MTBE. The solution was then cooled to 5°C at a rate of 0.2 °C / min. The crystallized product was then stirred for a further 15 minutes. The final slurry was filtered, and the resulting material was vacuum-dried at 40°C for approximately 16 hours.

[0186] Figure 4 shows the XRPD spectra of the substances produced by the above procedure. Figure 4A shows the XRPD spectrum of the substance from the first sample (i.e., after the addition of approximately 30 ml of MTBE), and Figure 4B shows the XRPD spectrum of the substance from the second sample (i.e., after the addition of approximately 60 ml of MTBE). Figure 4C shows the XRPD spectrum of the dried final product. Example 5: Synthesis of Fumagyl-6-yl N-(Trans-4-aminocyclohexyl)carbamatebenzenesulfonate and crystallization from ethyl acetate

[0187] Fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate was prepared as shown below. The procedure described in Table 17 below demonstrates that the compound of formula I can be directly purified without first acidifying the free base of formula I and extracting it into the aqueous layer, as described in Example 1, before extracting the free base returned to the organic phase by basicizing the aqueous layer. [Table 20]

[0188] As shown in Table 17 above, the water content in step 18 is maintained at 1–1.3%. While not intended to imply theoretical constraints, maintaining the water content within this range helps to promote the direct crystallization of the compound of formula I from ethyl acetate with good purity (e.g., over 95%). As shown in step 29, this method increased the yield by approximately 10–15% compared to the method of Example 1.

[0189] The quantities of the substances used in the above procedure are shown in Table 18 below. Table 18. Amount of substance in the synthesis of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate [Table 21]

[0190] As shown in Table 18, benzenesulfonic acid monohydrate (i.e., approximately 12-13% water and approximately 1-2% impurities) was used in step 20. Although not constrained by theory, it was found that formula I was obtained with the highest yield and highest purity when approximately 0.78-0.8 equivalents of benzenesulfonic acid were used.

[0191] While not intended to imply theoretical constraints, the trituration and washing of ethyl acetate in steps 26 and 27 were performed to reduce the water content to an acceptable level before drying, minimizing the risk of water splitting of formula I. In-Process Control for the Synthesis of Fumagillol-6-yl N-(trans-4-aminocyclohexyl)carbamate Benzenesulfonate

[0192] The HPLC conditions used in the above examples are specified below: [Table 22]

[0193] Table 19 below shows the results under in-process control as high performance liquid chromatography (HPLC) measurements for various batches of fumagillol-6-yl N-(trans-4-aminocyclohexyl)carbamate benzenesulfonate (Formula I). Formula I had a retention time of 16.18 minutes and a relative retention time of 1. Various impurities including Compound 2 and Compound 4 were defined by the retention times in Table 19.

[0194] As shown in Table 19, Lots 1-3 are control standards. Lot 1 was manufactured by the method of Example 1. Lots 2 and Lot 3 are comparative batches, and Lot 3 was recrystallized using methanol and methyl tert-butyl ether.

[0195] Lots 4-10 are in-process controls measured during the process described in Example 5 above. Lot 4 gives HPLC data taken 1 hour after Step 11 in Table 17. Lot 5 gives HPLC data after drying with Na2SO4 in Step 17. Lot 6 gives HPLC data after filtration in Step 25. Lot 7 gives HPLC data after washing Formula I once with ethyl acetate. Lot 8 gives HPLC data after washing Formula I twice with ethyl acetate. Lot 9 gives HPLC data after filtering and drying Formula I for 10 hours. Lot 10 gives HPLC data after filtering and drying Formula I for 24 hours. As described in the analysis of Lot 10, the purity of Formula I after 24 hours of drying was the same as that of Lot 1 manufactured by the method of Example 1 without recrystallization from methanol / methyl tert-butyl ether. [Table 23-1] [Table 23-2] Example 7 - Inhibition of hematological malignancy cell lines of formula I

[0196] The maximum median inhibitory concentration (IC) of formula I as a single agent against a panel of human hematological tumor cell lines. 50 The following was investigated: All cell lines were grown in their respective appropriate growth media supplemented with 5-10% FBS and cultured at 37°C in a 5% CO2 atmosphere. Single-drug research Cell viability assay - 72 hours of continuous exposure

[0197] 1.0 hour: Cells were seeded in 200 μL of growth medium in a 96-well microtiter plate. The cells were incubated in a humidified incubator at 37°C for 24 hours.

[0198] 2.24 hours: The drug dose of the test agent was obtained using an HPD300 digital dispenser. Briefly, the digital dispenser added the test agent in the exact volume of DMSO solvent to the culture medium containing the wells. The control wells were given the same volume of DMSO as the test agent wells. After drug administration, the cells were exposed in a humidified incubator at 37°C for 72 hours at the concentrations and dilutions listed in Table 20.

[0199] 3. 72 hours: After 72 hours of exposure, 100 μl of a 1:1 mixture of sterile water and CellTiter-Glo® reagent was added to each well. The plate was incubated at room temperature for 60 minutes. After incubation, luminescence was recorded using a light meter. The results are shown in Table 21 and Figures 15A, 15B, 15C, and 15D below. [Table 24] [Table 25] endpoint

[0200] I C 50 Decision: Data are expressed as the cell proliferation rate of the untreated (vehicle) control, calculated from the luminescence signal. Cell viability was determined by dividing the average luminescence value of the test drug by the average luminescence value of the untreated control. Inhibitory concentrations of the test drug and control were estimated by curve fitting of the data using nonlinear regression analysis with Prism 6 software (GraphPad Software, Inc.).

[0201] Evenly While this disclosure has been described in relation to the specific embodiments described above, many alternatives, modifications, and other variations will be apparent to those skilled in the art. All such alternatives, modifications, and variations will fall within the intent and scope of this disclosure.

Claims

1. A method for producing crystals of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamate 1-hydroxy-2-naphthoate, characterized by peaks at about 5.6, about 8.9, about 11.5, about 12.1, about 15.4, and about 20.9°2θ as measured by X-ray powder diffraction, comprising crystallizing fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamate 1-hydroxy-2-naphthoate from acetonitrile, ethyl acetate, methyl ethyl ketone, or tetrahydrofuran.

2. A method for producing a crystal according to claim 1, further characterized by peaks at approximately 5.6, approximately 8.9, approximately 11.5, approximately 12.1, approximately 15.4, approximately 15.9, approximately 19.7, approximately 20.9, and 23.3°2θ, as measured by X-ray powder diffraction.

3. A method for producing a crystal according to claim 1, further characterized by peaks at approximately 5.6, 8.9, 11.5, 12.1, 14.6, 15.4, 15.9, 17.4, 18.1, 19.7, 20.9, and 23.3°2θ, as measured by X-ray powder diffraction.

4. The method for producing a crystal characterized by the X-ray powder diffraction pattern shown in Figure 10A, 10B, 10C, or 10D, according to claim 1.

5. A method for producing a crystal according to claim 1, further characterized by the onset of an endothermic event at approximately 181°C and a peak at approximately 186°C, and an exothermic event at approximately 191°C, as measured by thermogravimetric analysis / differential thermal analysis.

6. The method for producing a crystal according to claim 1, characterized by the onset of an endothermic event at approximately 182°C and a peak at approximately 186°C, as measured by differential scanning calorimeter (DSC).

7. A composition for treating any disease selected from B-cell lymphoma, mantle cell lymphoma, diffuse large B-cell lymphoma, and myeloma, comprising crystals of fumagyl-6-yl N-(trans-4-aminocyclohexyl)carbamatebenzenesulfonate, characterized by an X-ray powder diffraction pattern using Cu Kα irradiation with peaks at approximately 6.0, 9.0, 12.3, 12.5, 13.8, 16.1, 18.0, 20.0, and 25.8 °2θ.

8. The composition according to claim 7, wherein the disease is B-cell lymphoma.

9. The composition according to claim 7, wherein the disease is mantle cell lymphoma.

10. The composition according to claim 7, wherein the disease is diffuse large B-cell lymphoma.

11. The composition according to claim 7, wherein the disease is myeloma.

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