Crystalline forms of tegavivint, methods of preparation, and uses thereof

Abstract

A formulation of (9E,10E)-2,7-bis((3,5-dimethylpiperidin-1-yl)sulfonyl)anthracene-9,10-dione dioxime (tegavivint) with long-term physical stability is provided. Crystalline forms of tegavivint, pharmaceutical compositions containing the crystalline forms, processes for preparing the crystalline forms, and methods of use therefor are provided.

Description

[Technical Field]

[0001] The present invention relates to a crystalline form of tegavivint, also known as (9E,10E)-2,7-bis((3,5-dimethylpiperidin-1-yl)sulfonyl)anthracene-9,10-dione dioxime, pharmaceutical compositions containing the crystalline form, processes for preparing the crystalline form, and methods of use thereof. [Background technology]

[0002] Cancer is the second leading cause of death in the United States. It presents complex challenges for the development of new treatments. Cancer is characterized by the abnormal growth and metastasis of malignant cells that undergo a series of genetic changes that result in the growth of a tumor mass.

[0003] Beta-catenin (β-catenin) is part of a complex of proteins that constitute adherens junctions (AJs). AJs are required for the generation and maintenance of epithelial cell layers by regulating cell growth and cell-cell adhesion. β-catenin also anchors the actin cytoskeleton and may be involved in the transmission of contact inhibition signals that cause the cessation of cell division when the epithelial sheet is completed.

[0004] The Wnt / β-catenin pathway has been shown to play a role in cancer. Abnormal β-catenin signaling plays an important role in tumorigenesis. In particular, it is estimated that colorectal cancer has more than 80% mutations in the β-catenin pathway, resulting in deregulated oncogenic signaling. Abnormal β-catenin signaling has been shown to be involved in various types of cancer, including, but not limited to, melanoma, breast cancer, lung cancer, colon cancer, liver cancer, gastric cancer, myeloma, multiple myeloma, chronic myelogenous leukemia, chronic lymphocytic leukemia, T-cell non-Hodgkin's lymphoma, colorectal cancer, and acute myeloid leukemia (AML). Furthermore, abnormal Wnt / β-catenin signaling has been found in many other disorders, including, but not limited to, osteoporosis, osteoarthritis, polycystic kidney disease, diabetes, schizophrenia, vascular disease, heart disease, hyperproliferative disorders, neurodegenerative diseases, and fibrotic diseases, including idiopathic pulmonary fibrosis (IPF), Dupuytren's contracture, and nonalcoholic steatohepatitis (NASH). Myeloproliferative neoplasms (MPNs) are a group of closely related hematologic malignancies in which the body's blood-producing bone marrow cells develop and function abnormally. The three major myeloproliferative neoplasms are polycythemia vera (PV), essential thrombocythemia (ET), and primary myelofibrosis (PMF). JAK2 gene mutations are found in most PV patients and in 50% of ET and PMF patients. The β-catenin pathway is often activated in MPNs and is required for the survival of these cells.

[0005] Tegavivint and related compounds are described, for example, in U.S. Patent No. 8,129,519. Tegavivint has the following structural formula: [ka] The chemical name is (9E,10E)-2,7-bis((3,5-dimethylpiperidin-1-yl)sulfonyl)anthracene-9,10-dionedioxime.

[0006] The molecular formula of Tegavivin is C 28 H 36 It is N4O6S2.

[0007] The molecular weight of Tegavivint is 588.20763 amu.

[0008] The small-scale chemical synthesis of tegavivint is disclosed in U.S. Patent No. 8,129,519. The drug substance / active pharmaceutical ingredient (API) has good chemical and physical stability. However, there are significant concerns about the physical stability of nanoparticulate formulations of tegavivint over time, which may manifest as crystal growth (Oswald ripening) or polymorphic transformation, which may result in an increase in the number of large particles or the generation of undesirable particle morphologies during long-term storage of the formulation. Therefore, there remains a need to conduct crystallographic studies to explore suitable / related polymorphs of tegavivint that may be amenable to disruption and formulation development to obtain formulations with good long-term physical stability. The present invention advantageously addresses this need. [Prior art documents] [Patent documents]

[0009] [Patent Document 1] U.S. Patent No. 8,129,519 Summary of the Invention

[0010] This application discloses an invention that addresses the aforementioned problems and needs by providing a crystalline single polymorphic form of tegavivint, referred to throughout this application as Form IV. Current formulations of tegavivint are nanosuspensions produced using a milling process. While Form I (BC-2059 obtained directly from chemical synthesis) is currently utilized as the starting material for the milling process, and the final product obtained from milling is a nanosuspension of Form I, the inventors have unexpectedly discovered that there are certain advantages to utilizing Form IV (compared to Form I) as the starting material for the milling process to prepare a nanosuspension of tegavivint.

[0011] The main advantage is that Form IV is sufficiently unstable that it converts to Form I when crushed at high temperature (60°C). Therefore, the system undergoes complete solvent-mediated recrystallization from Form IV to Form I. Because Form I crystals grow "bottom-up" as they are crushed, the possibility of obtaining larger uncrushed crystals will be significantly reduced. In other words, utilizing Form IV as the starting material is beneficial because it will eventually convert to Form I, and recrystallization from Form IV will only yield Form I crystals. Therefore, promoting a suspension with a single polymorph produced by crushing at high temperature will improve the stability of the suspension.

[0012] Thus, in one embodiment, the present invention provides a crystalline form of tegavivint designated Form IV, having an X-ray powder diffraction pattern (XRPD) comprising diffraction peaks having °2θ angles selected from the group consisting of 5.0±0.2°, 7.5±0.2°, 7.7±0.2°, 10.2±0.2°, 14.8±0.2°, 15.2±0.2°, 15.4±0.2°, 18.0±0.2°, 20.0±0.2°, 20.5±0.2°, and 22.2±0.2°.

[0013] In one embodiment, Form IV may be monocrystalline.

[0014] In one embodiment, Form IV is a trihydrate.

[0015] In another embodiment, Form IV has an endothermic peak at about 115.9°C.

[0016] In another embodiment, Form IV has an exothermic peak onset at about 147.1°C.

[0017] In yet another embodiment, the exothermic decomposition of Form IV begins at about 280°C.

[0018] In yet another embodiment, the present invention provides a nanosuspension of tegavivint, prepared by a process comprising using Form IV as a starting material and crushing Form IV at a temperature of about 40°C to about 60°C, most preferably about 60°C.

[0019] In one embodiment, if the crushing step is carried out at a temperature below about 60°C, the nanosuspension must further undergo an annealing step at 60°C or above.

[0020] In another embodiment of the present invention, a pharmaceutical composition is provided for use in the method, comprising a stable nanosuspension of Form I prepared using Form IV as a starting material and a pharmaceutically acceptable excipient.

[0021] In another embodiment of the present invention, there is provided herein a method for preventing, treating, or ameliorating cancer or tumor metastasis in a mammal in need thereof, comprising administering to said mammal an effective amount of a composition of the present invention. [Brief explanation of the drawings]

[0022] The patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the U.S. Patent and Trademark Office upon request and payment of the necessary fee.

[0023] [Figure 1] 1 shows the X-ray powder diffraction (XRPD) pattern of Tegavivint Form I. [Figure 2] 1 shows the differential scanning calorimetry (DSC) and thermogravimetric analysis (TGA) curves of Form I. [Figure 3] Polarized light microscope (PLM) image of Form I is shown. [Figure 4] 1 shows an XRPD pattern overlay of Form II formulation. [Figure 5] 1 shows an XRPD pattern overlay of Form III formulation. [Figure 6]1 shows the DSC and TGA curves of Form III. [Figure 7] PLM image of Form III is shown. [Figure 8] 1 shows an XRPD pattern overlay of a Form III sample after drying. [Figure 9A] 1 shows the XRPD pattern of a Form IV sample. [Figure 9B] 1 shows the DSC and TGA curves of Form IV. [Figure 9C] 1 shows the dynamic vapor sorption (DVS) profile of Form IV. [Figure 9D] PLM images of Form IV are shown. [Figure 9E] 1 shows the variable temperature X-ray powder diffraction (VT-XRPD) profile of Form IV. [Figure 9F] 1 shows VT-XRPD and XRPD pattern overlays of a Form IV sample after exposure to ambient conditions. [Figure 10] 1 shows the XRPD pattern of Form V. [Figure 11] 1 shows the XRPD pattern of Form VI. [Figure 12A] The XRPD pattern of the amorphous sample is shown. [Figure 12B] 1 shows the modulated DSC (mDSC) curve of an amorphous sample. [Figure 13A] 1 shows the XRPD pattern of the solid obtained from the slurry competition in water. [Figure 13B] Figure 1 shows the XRPD pattern of the solid obtained from the slurry competition in ACN / water (1:1, v / v). [Figure 13C] Figure 1 shows the XRPD pattern of the solid obtained from the slurry competition in ACN / water (1:3, v / v). [Figure 14A] 1 is a particle size distribution (PSD) plot of Form I samples crushed at 5° C. [Figure 14B] 1 is a PSD plot of Form I samples crushed at room temperature. [Figure 14C] 1 is a PSD plot of Form I samples crushed at 60° C. [Figure 14D]1 is a PSD plot of Form IV samples crushed at 5° C. [Figure 14E] 1 is a PSD plot of Form IV sample crushed at room temperature. [Figure 14F] 1 is a PSD plot of Form IV samples crushed at 60° C. [Figure 14G] PLM images of Form I and Form IV. [Figure 14H] 10 is a PLM image of Form I sample crushed at 5° C. [Figure 14I] 10 is a PLM image of Form I sample fractured at room temperature. [Figure 14J] 16 is a PLM image of Form I sample fractured at 60°C. [Figure 14K] 16 is a PLM image of Form IV sample fractured at 5° C. [Figure 14L] 13 is a PLM image of Form IV sample fractured at room temperature. [Figure 14M] 16 is a PLM image of Form IV sample fractured at 60° C. [Figure 14N] 1 shows an XRPD pattern overlay of a Form I sample crushed at 5° C. [Figure 14O] 1 shows an XRPD pattern overlay of a Form I sample crushed at room temperature. [Figure 14P] 1 shows an XRPD pattern overlay of a Form I sample crushed at 60° C. [Figure 14R] 1 shows an XRPD pattern overlay of a Form IV sample crushed at 5° C. [Figure 14S] 1 shows an XRPD pattern overlay of a Form IV sample crushed at room temperature. [Figure 14T] 1 shows an XRPD pattern overlay of a Form IV sample crushed at 60° C. DETAILED DESCRIPTION OF THE INVENTION

[0024] The present invention relates to a crystalline form of tegavivint. In particular, the present invention relates to a crystalline form of tegavivint designated Form IV, pharmaceutical compositions containing the crystalline form, processes for preparing the crystalline form, and methods of use thereof.

[0025] In one embodiment, the crystalline form of tegavivint is referred to as Form IV and has an X-ray powder diffraction pattern (XRPD) comprising diffraction peaks having °2θ angle values ​​independently selected from the group consisting of 5.0±0.2°, 7.5±0.2°, 14.8±0.2°, 15.2±0.2°, 15.4±0.2°, 20.0±0.2°, and 22.2±0.2°.

[0026] In one embodiment, Form IV has an XRPD comprising diffraction peaks having °2θ angle values ​​independently selected from the group consisting of 5.0±0.2°, 7.5±0.2°, 7.7±0.2°, 14.8±0.2°, 15.2±0.2°, 15.4±0.2°, 20.0±0.2°, and 22.2±0.2°.

[0027] In one embodiment, Form IV has an XRPD comprising diffraction peaks having °2θ angle values ​​independently selected from the group consisting of 5.0±0.2°, 7.5±0.2°, 7.7±0.2°, 10.2±0.2°, 14.8±0.2°, 15.2±0.2°, 15.4±0.2°, 20.0±0.2°, and 22.2±0.2°.

[0028] In another embodiment, Form IV has an XRPD comprising diffraction peaks having °2θ angle values ​​independently selected from the group consisting of 5.0±0.2°, 7.5±0.2°, 7.7±0.2°, 10.2±0.2°, 14.8±0.2°, 15.2±0.2°, 15.4±0.2°, 18.0±0.2°, 20.0±0.2°, and 22.2±0.2°.

[0029] In another embodiment, Form IV has an XRPD comprising diffraction peaks having °2θ angle values ​​independently selected from the group consisting of 5.0±0.2°, 7.5±0.2°, 7.7±0.2°, 10.2±0.2°, 14.8±0.2°, 15.2±0.2°, 15.4±0.2°, 18.0±0.2°, 20.0±0.2°, 20.5±0.2°, and 22.2±0.2°.

[0030] In another embodiment, Form IV has an XRPD pattern substantially as shown in Figure 9A.

[0031] In another embodiment, Form IV is characterized by differential scanning calorimetry (DSC) as having an endotherm with a peak maximum at about 115.9°C.

[0032] In another embodiment, Form IV is characterized by DSC as having an exothermic peak onset at about 147.1°C.

[0033] In one embodiment, Form IV is characterized by having a DSC thermogram substantially as shown in Figure 9B.

[0034] In one embodiment, Form IV is a trihydrate isolated from a solvent with high water activity. The crystalline form is needle-like. The trihydrate thermally dehydrates at a halfway point of dehydration at about 60°C.

[0035] In yet another embodiment, the present invention provides a nanosuspension of tegavivint, wherein the nanosuspension is prepared by a process comprising using Form IV as a starting material and crushing Form IV at a temperature of about 40°C to about 60°C, most preferably about 60°C.

[0036] In one embodiment, if the crushing step is carried out at a temperature below about 60°C, the nanosuspension must further undergo an annealing step at 60°C or above.

[0037] In another embodiment of the present invention, there is provided a pharmaceutical composition for use in the method, comprising a stable nanosuspension of Form I prepared using Form IV as a starting material, and a pharmaceutically acceptable excipient.

[0038] In another embodiment of the present invention, provided herein is a method for preventing, treating, or ameliorating cancer or tumor metastasis in a mammal in need thereof, comprising administering to said mammal an effective amount of a composition of the present invention.

[0039] The crystalline form of tegavivint may be formulated by any method known in the art and prepared for administration by any route, including, but not limited to, parenteral, oral, sublingual, transdermal, topical, nasal, intratracheal, or rectal. In certain embodiments, the crystalline form of tegavivint is administered intravenously in a hospital. In one embodiment, it may be administered orally.

[0040] The characteristics of the carrier depend on the route of administration. As used herein, the term "pharmaceutically acceptable" refers to a non-toxic material that is compatible with a biological system, such as a cell, cell culture, tissue, or organism, and does not interfere with the effectiveness of the biological activity of the active ingredient. Thus, in addition to the inhibitor, the composition may contain diluents, fillers, salts, buffers, stabilizers, solubilizers, and other materials known in the art. The preparation of pharmaceutically acceptable formulations is described, for example, in Remington's Pharmaceutical Sciences, 18th Edition, ed. A. Gennaro, Mack Publishing Co., Easton, Pa., 1990.

[0041] Pharmaceutical compositions containing crystalline forms of tegavivint may be used in the methods of use described herein.

[0042] The active compound is included in a pharmaceutically acceptable carrier or diluent in an amount sufficient to deliver a therapeutically effective amount to the patient without causing serious toxic effects to the treated patient. The effective dosage range of the pharmaceutically acceptable derivative can be calculated based on the weight of the parent compound to be delivered. If the derivative exhibits activity in itself, the effective dosage can be estimated using the weight of the derivative as described above, or by other means known to those skilled in the art.

[0043] In some embodiments of any of the methods described herein, prior to treatment with a composition or method of the invention, the patient may have been treated with one or more of chemotherapy, targeted anti-cancer agents, radiation therapy, and surgery, optionally where the previous treatment was unsuccessful; and / or the patient has undergone surgery, optionally where the surgery was unsuccessful; and / or the patient has been treated with a platinum-based chemotherapy agent, optionally where the patient has previously been determined to be unresponsive to treatment with the platinum-based chemotherapy agent; and / or the patient has been treated with a kinase inhibitor, optionally where the previous treatment with the kinase inhibitor was unsuccessful; and / or the patient has been treated with one or more therapeutic agents.

[0044] definition Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. All patents, patent applications, and publications mentioned herein are incorporated by reference.

[0045] As used herein, "tegavivint" refers to (9E,10E)-2,7-bis((3,5-dimethylpiperidin-1-yl)sulfonyl)anthracene-9,10-dione dioxime.

[0046] As used herein, the term "Form IV" or "crystalline Form IV" when used alone refers to crystalline Form IV of (9E,10E)-2,7-bis((3,5-dimethylpiperidin-1-yl)sulfonyl)anthracene-9,10-dionedioxime.

[0047] As used herein, the terms "subject," "individual," or "patient" are used interchangeably and refer to any animal, including mammals such as mice, rats, other rodents, rabbits, dogs, cats, pigs, cows, sheep, horses, primates, and humans. In some embodiments, the patient is human. In some embodiments, the subject is experiencing and / or exhibiting at least one symptom of the disease or disorder being treated and / or prevented. In some embodiments, the subject is suspected of suffering from a multi-tyrosine kinase-associated cancer.

[0048] As used herein, a "therapeutically effective amount" of a crystalline form of tegavivint is an amount sufficient to alleviate or in any way relieve the symptoms, or to halt or reverse the progression of a condition, or to negatively regulate or inhibit the activity of a multi-tyrosine kinase. Such an amount may be administered in a single dose or according to a regimen such that it is effective.

[0049] As used herein, "treatment" means any manner in which the symptoms or pathology of a condition, disorder or disease are ameliorated or otherwise beneficially altered. Treatment also includes any pharmaceutical use of the compositions herein.

[0050] As used herein, amelioration of symptoms of a particular disorder by administration of a particular pharmaceutical composition refers to any permanent or temporary, sustained or transient relief that may result from or be associated with administration of the composition.

[0051] As used herein, when used to modify a numerically defined parameter (e.g., a dose of a crystalline form of tegavivint or a pharmaceutically acceptable salt thereof detailed herein, or a length of treatment time described herein), the term "about" means that the parameter can vary by up to 10% below or above the numerical value stated for that parameter. For example, a dose of about 5 mg / kg can vary from 4.5 mg / kg to 5.5 mg / kg. When used at the beginning of a list of parameters, "about" is meant to modify the respective parameter. For example, about 0.5 mg, 0.75 mg, or 1.0 mg means about 0.5 mg, about 0.75 mg, or about 1.0 mg. Similarly, about 5% or more, 10% or more, 15% or more, 20% or more, and 25% or more mean about 5% or more, about 10% or more, about 15% or more, about 20% or more, and about 25% or more.

[0052] As used herein, the term "about" when used in reference to XRPD peak positions refers to the inherent variability of the peak depending on the calibration of the instrument, the process used to prepare the crystalline forms of the present invention, the age of the crystalline form, and the type of instrument used for the analysis. The variability of the instrument used for the XRPD analysis is about ±0.2 o It was 2θ.

[0053] As used herein, when used in reference to the appearance of a DSC endothermic peak, the term "about" refers to the inherent variability of the peak depending on the calibration of the instrument, the method used to prepare the samples of the present invention, and the type of instrument used for the analysis. The variability of the instrument used for the DSC analysis was about ±2°C.

[0054] General Method The general methods outlined below were used in the illustrated examples unless otherwise stated.

[0055] The crystalline forms of the present invention may be prepared using a variety of methods well known to those skilled in the art, including crystallization or recrystallization from a suitable solvent or sublimation. A wide variety of techniques can be employed, including evaporation of a water-miscible or water-immiscible solvent, seeding with crystals in a supersaturated solvent mixture, reducing the temperature of the solvent mixture, or freeze-drying the solvent mixture for crystallization or recrystallization.

[0056] In the present invention, crystallization can be carried out with or without crystal seeds. Crystal seeds can be derived from any previous batch of the desired crystalline form. The addition of crystal seeds may not affect the preparation of the crystalline form in the present invention.

[0057] Samples were withdrawn after completion of the isotherm and reanalyzed by XRPD. Abbreviations and Acronyms [Table 1]

[0058] The following examples are intended to illustrate further specific embodiments of the present invention and are not intended to limit the scope of the invention. [Example]

[0059] Example 1 Investigation of Tega Vivint Form I This example describes the investigation of Form I of Tegavivint.

[0060] The starting material (Tegavivint Form I) was characterized by XRPD, TGA, DSC, and PLM. The XRPD pattern, shown in Figure 1, indicated that the starting material was crystalline and confirmed to be Form I. The TGA and DSC curves are shown in Figure 2. A weight loss of 0.4% was observed up to 150 °C in the TGA curve, and the DSC results showed no melting endotherm before decomposition. Based on the characterization results, Form I was inferred to be anhydrous. The PLM image, shown in Figure 3, showed irregular particulates with partial aggregation for the Form I sample.

[0061] Example 2 Preparation of Tegavivint Form II Attempts to prepare Form II of tegavivint were carried out under four conditions, with detailed results shown in Figure 4 and Table 1. The results suggested that Form II was very difficult to reconstitute or was metastable.

[0062] [Table 2]

[0063] Example 3 Preparation of Tegavivint Form III Form III samples (803759-03-A and 803759-05-A1) were prepared via anti-solvent addition in MeOH / HO and EtOH / HO systems, and the XRPD results are shown in Figure 5. The TGA / DSC results for Form III (803759-05-A1) are shown in Figure 6. An 8.2% weight loss by 100°C was observed on TGA. One endotherm at 64.5°C and one exotherm at 158.6°C were observed before decomposition on DSC. The PLM image shown in Figure 7 showed needles and fine particles with aggregation for the Form III sample (803759-05-A1).

[0064] Form III may be isomorphous because Form III samples were obtained from different solvent systems and converted to Form IV after drying under vacuum at room temperature or to Form V after exposure to ambient conditions at room temperature (Figure 8).

[0065] Example 4 Preparation of Tegavivint Form IV A sample of Form IV (803759-13-B) was reprepared on a 2 g scale, the detailed procedure being as follows:

[0066] 1. Weigh 2.0 g of 803759-01-A sample into a 1 L reactor. 2. Add 200 mL of EtOH and stir at 300 rpm at room temperature to obtain a clear solution. 3. Inject 100 mL of water. 4. 90.2 mg of Form IV seeds was added and the suspension was observed. 5. Inject 100 mL of water over 1 hour. 6. Keep the slurry for 2 hours. 7. Filter and test the XRPD of the wet solid. 8. Transfer the wet solids to a 1 L reactor and inject with 200 mL of water and slurry overnight. 9. Filter and vacuum dry for 4 hours to give 1.9 g of solid (803759-13-B, Form IV).

[0067] XRPD results for Form IV are shown in Figure 9A. TGA / DSC results are shown in Figure 9B. In TGA, an 8.4% weight loss was observed up to 150°C. In DSC, one endotherm at 115.9°C (peak) and one exotherm at 147.1°C (exotherm) were observed before decomposition. DVS results, shown in Figure 9C, indicated the following: 1) Two platforms were observed, indicating the presence of two potential hydrate forms. 2) A 9.1% water uptake was observed at 25°C / 80% RH, consistent with the TGA weight loss of Form IV. The PLM image displayed in Figure 9D indicated that needle-like particles were observed for the Form IV sample (803759-13-B).

[0068] VT-XRPD testing was employed to further investigate Form IV, and the results, shown in Figures 9E and 9F, indicated that 1) the Form IV sample partially converted to a new form at 30°C under N2, which was designated Form VI; 2) pure Form VI was observed after heating to 75°C and 120°C; 3) Form VI was still observed after cooling to 30°C; and 4) after exposure to ambient conditions, Form VI rapidly converted to Form IV. Therefore, Form IV is believed to be a hydrate.

[0069] Example 5 Preparation of Tegavivint Form V Form V sample (803759-03-A April 22) was obtained by drying Form III sample (803759-03-A) at ambient conditions, and the XRPD pattern is displayed in Figure 10. Due to limited solids, TGA and DSC data were not collected.

[0070] Example 6 Preparation of Tegavivint Form VI During VT-XRPD testing of the Form IV sample (803759-02-B), a Form VI sample (803759-02-B_N2 back_30.0°C) was obtained. The XRPD pattern is shown in Figure 11. After exposure to ambient conditions, Form VI rapidly converted to Form IV (Figure 9F). Because Form VI was unstable under ambient conditions, no further characterization data was collected on Form VI, and it was assumed to be an anhydrate.

[0071] Example 7 Preparation of amorphous form of tegavivint An amorphous sample (803759-04-B3 dried) was prepared by adding an antisolvent in a DMSO / HO system and drying under vacuum at room temperature. The XRPD pattern is shown in Figure 12A. The mDSC results shown in Figure 12B indicated that the Tg of the amorphous sample was 65.0 °C (medium temperature).

[0072] Example 8 Slurry competition experiment To determine the most stable form under high water activity at room temperature, slurry competitions of Forms I, III, IV, and V were carried out in three solvent systems with high water activity at room temperature: water, ACN / HO (1:1, v / v), and ACN / HO (1:3, v / v).

[0073] The detailed steps are as follows: 1) Prepare saturated solutions of Form I sample (803759-01-A) in three solvent systems. 2) Add approximately 10 mg of each form to the corresponding 1 mL of saturated solution. 3) The wet solids after 1 and 6 days were slurried and examined by XRPD.

[0074] The results are shown in Table 2 and Figures 13A, 13B and 13C. The results showed the following: 1) Form IV was obtained from an aqueous system. 2) A new form was obtained from the ACN / H2O (1:1, v / v) system, which was presumed to be an ACN solvate. 3) Form I was obtained from the ACN / H2O (1:3, v / v) system.

[0075] [Table 3]

[0076] Example 9 Ball milling experiment Previously, Form I was crushed to a very small particle size and began to convert to Form IV in aqueous suspension during storage, with unacceptable particle size growth. Therefore, Form I and Form IV were ball milled to evaluate form stability and particle size growth. The detailed ball milling procedure is as follows: 1. Form I and Form IV samples are separately suspended in 1% aqueous Poloxamer 188 solution (50 mg / mL). 2. Add approximately 12 mL of the suspension to a 50 mL tube containing the crushing beads (the volume of the crushing beads is approximately 30 mL). After adding 12 mL of the suspension, the liquid surface just covered the beads. 3. Rotate the 50 mL tube (containing the beads and suspension) at 30 rpm at 5°C, room temperature, and 60°C. Using a 4.1-mL syringe, approximately 0.8 mL of the suspension is sampled at 1, 2, 4, and 24 hours. 5. Test the suspension for XRPD, PSD and PLM.

[0077] The results shown in Table 3 and Figures 14A-14T indicated the following: 1) The morphological transformation of Form IV was observed after 24 hours at room temperature and after 2 hours at 60°C. 2) The particle size decreased during crushing. 3) Aggregation was observed for Form I at 5°C and for Form IV at 5°C and at room temperature after 24 hours. Therefore, ball milling of Form I samples at elevated temperature (60°C) was recommended to reduce particle size. Additionally, the expected result is that crushing Form I at temperatures above 60°C will prevent the formation of undesirable Form IV crystal seeds and result in a highly crystalline crushed material that is annealed, free of energetic particles, and free of amorphous material. Conversion to Form I was confirmed by ball milling Form IV at elevated temperature (60° C.).

[0078] [Table 4] [Table 5]

[0079] In summary, five crystalline forms of Tegavivint and an amorphous sample were obtained, as summarized in Table 4.

[0080] [Table 6]

[0081] appendix Apparatus and method XRPD For XRPD analysis, PANalytical X'Pert 3 An X-ray powder diffractometer was used, and the XRPD parameters used are listed in Table 5.

[0082] [Table 7]

[0083] TGA, DSC and mDSC TGA data were collected using a TA Instruments TA Discovery 5500 / Q5000TGA. DSC and mDSC were performed using a TA Instruments TA Discovery 2500 DSC. The detailed parameters used are shown in Tables 6 and 7.

[0084] [Table 8]

[0085] [Table 9]

[0086] DVS DVS was measured by a DVS Intrinsic manufactured by SMS (Surface Measurement Systems). Relative humidity at 25°C was calibrated against the decomposition points of LiCl, Mg(NO3)2, and KCl. The DVS test parameters are listed in Table 8.

[0087] [Table 10]

[0088] PLM PLM images were captured using a Carl Zeiss (Germany) Axio Scope A1 microscope.

[0089] PSD A Microtrac S3500 equipped with an SDC (sample delivery controller) was used for the PSD test, and the method is shown in Table 9.

[0090] [Table 11]

[0091] While the invention has been described in relation to particular embodiments thereof, it will be understood that it is capable of further modifications, and this application is intended to cover generally any variation, use, or adaptation of the invention in accordance with its principles, including departures from the present disclosure within known or customary practice in the art to which this invention pertains, provided the essential features described herein are applicable and in accordance with the scope of the appended claims.

Claims

1. The following formula: 【Chemistry 1】 A crystal of form IV of a compound having, wherein the crystal of form IV exhibits endothermic properties before decomposition in differential scanning calorimetry (DSC), and the endothermic properties have a peak maximum value of 115.9°C.

2. The crystal according to claim 1, wherein the crystal of form IV further generates heat before decomposition in differential scanning calorimetry (DSC).

3. The crystal according to claim 2, wherein the exothermic reaction has a peak onset temperature of 147.1°C.

4. The crystal according to any one of claims 1 to 3, wherein the crystal of form IV further has exothermic decomposition that begins at 280°C.

5. The crystal according to any one of claims 1 to 4, wherein the crystal of form IV further has a weight loss of 8.4% up to 150°C in thermogravimetric analysis (TGA).

6. The crystal according to any one of claims 1 to 5, wherein the crystal of form IV further has a moisture absorption of 9.1% at 25°C and a relative humidity (RH) of 80% in dynamic water vapor sorption (DVS).

7. The crystal according to any one of claims 1 to 6, wherein the crystal of form IV has an X-ray powder diffraction pattern (XRPD) including diffraction peaks having °2θ angular values ​​of 14.8 ± 0.2°, 15.2 ± 0.2°, and 15.4 ± 0.2°.

8. The crystal according to any one of claims 1 to 6, wherein the crystal of the above-mentioned form IV has an X-ray powder diffraction pattern (XRPD) including diffraction peaks having °2θ angular values ​​consisting of 5.0±0.2°, 7.5±0.2°, 14.8±0.2°, 15.2±0.2°, 15.4±0.2°, 20.0±0.2°, and 22.2±0.2°.

9. The crystal of the above-mentioned form IV has an X-ray powder diffraction pattern (XRPD) including diffraction peaks having °2θ angular values ​​consisting of 5.0±0.2°, 7.5±0.2°, 7.7±0.2°, 14.8±0.2°, 15.2±0.2°, 15.4±0.2°, 20.0±0.2°, and 22.2±0.2°, according to any one of claims 1 to 6.

10. The crystal of the above-mentioned form IV has an X-ray powder diffraction pattern (XRPD) including diffraction peaks having °2θ angular values ​​consisting of 5.0±0.2°, 7.5±0.2°, 7.7±0.2°, 10.2±0.2°, 14.8±0.2°, 15.2±0.2°, 15.4±0.2°, 20.0±0.2°, and 22.2±0.2°, according to any one of claims 1 to 6.

11. The crystal of the above-mentioned form IV has an X-ray powder diffraction pattern (XRPD) including diffraction peaks having °2θ angular values ​​consisting of 5.0±0.2°, 7.5±0.2°, 7.7±0.2°, 10.2±0.2°, 14.8±0.2°, 15.2±0.2°, 15.4±0.2°, 18.0±0.2°, 20.0±0.2°, and 22.2±0.2°, according to any one of claims 1 to 6.

12. The crystal of the above-mentioned form IV has an X-ray powder diffraction pattern (XRPD) including diffraction peaks having °2θ angular values ​​consisting of 5.0±0.2°, 7.5±0.2°, 7.7±0.2°, 10.2±0.2°, 14.8±0.2°, 15.2±0.2°, 15.4±0.2°, 18.0±0.2°, 20.0±0.2°, 20.5±0.2°, and 22.2±0.2°, according to any one of claims 1 to 6.

13. The crystal according to any one of claims 1 to 12, wherein the crystal of form IV is a hydrate.

14. The crystal according to claim 13, wherein the crystal of form IV is a trihydrate.

15. The crystal according to claim 14, wherein the trihydrate is thermally dehydrated, and the temperature at which the dehydration rate is halved is 60°C.

16. The crystal according to any one of claims 1 to 15, wherein the crystal form is needle-shaped.

17. As a starting material, use a crystal of form IV as described in any one of claims 1 to 16, Crushing the crystal of form IV at a temperature of 60°C. A method for preparing a nano-suspension of morph I of tegavivint, including the above.

18. To prepare the nanosuspension of claim 17, and Combining a therapeutically effective amount of the nano-suspension with a pharmaceutically acceptable excipient and / or diluent. A method for preparing a pharmaceutical composition, including [a specific compound / substance].

19. A method for preparing a pharmaceutical composition for treating or improving cancer or tumor metastasis in mammals requiring treatment or improvement of cancer or tumor metastasis, To prepare the nano-suspension of claim 17, and Combining a therapeutically effective amount of the nano-suspension with a pharmaceutically acceptable excipient and / or diluent. Methods that include...

20. The method according to claim 19, wherein the cancer is acute myeloid leukemia (AML).

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