Crystalline polymorphic forms of marimastat, amorphous forms of marimastat, methods for manufacturing the same, pharmaceutical compositions containing the same, and uses thereof

Abstract

The present invention relates to crystalline polymorphic forms I, II, and III of Marimastat, mixtures of crystalline polymorphic forms I and III of Marimastat, amorphous forms of Marimastat, and methods for preparing the same. The present invention also relates to a pharmaceutical composition comprising crystalline polymorphic forms I, II, or III of Marimastat, mixtures of crystalline polymorphic forms I and III of Marimastat, amorphous forms of Marimastat, or mixtures thereof. The present invention also relates to a pharmaceutical composition for use as a drug and for use in the prevention and treatment of diseases associated with the hyperactivity of extracellular matrix metalloproteinases, crystalline polymorphic forms I, II, or III of Marimastat, mixtures of crystalline polymorphic forms I and III of Marimastat, amorphous forms of Marimastat, or mixtures thereof.

Description

Technology Field

[0001] The present invention relates to a crystalline polymorphic form of marimastat, an amorphous form of marimastat, a method for manufacturing the same, a pharmaceutical composition containing the same, and the use of the same in the prevention and treatment of diseases associated with the hyperactivity of extracellular matrix metalloproteinases. Background Technology

[0002] Marimarstat or (2R,3S)-N1-((S)-3,3-dimethyl-1-methylamino-1-oxobutane-2-yl)-N4,3-dihydroxy-2-isobutylsuccinamide is a compound having the following structural formula:

[0003]

[0004] Marimastat is an inhibitor of extracellular matrix metalloproteinases (MMPs). Extracellular matrix metalloproteinases belong to a family of enzymes involved in the degradation of the extracellular matrix, which forms the tissue scaffold and enables cells to exchange information with the environment. Excessive activity of extracellular matrix metalloproteinases causes the destruction of the extracellular matrix, leading to impaired cellular function, which contributes to the development of many diseases. Excessive activity of extracellular matrix metalloproteinases occurs particularly in arthritis, periodontitis, atherosclerosis, and cardiovascular diseases [1, 2, 3]. Metalloproteinases play a significant role in the development and severity of vascular complications in diabetic patients [4]. They are believed to be involved in angiogenesis as well as cell differentiation, migration, and apoptosis [3]. Metalloproteinases play a significant role in carcinogenesis and metastasis. Their activity is correlated with the malignancy and metastatic tendency of tumors [1, 2, 3]. Increased extracellular matrix metalloproteinase activity is found in Alzheimer's disease, multiple sclerosis, amyotrophic lateral sclerosis, multiple myeloma, and malignant melanoma [10, 11]. It has been demonstrated that marimastat crosses the blood-brain barrier and exerts inhibitory effects on metalloproteinase activity in the brain. In mouse studies, marimastat reduced the duration and frequency of seizures in a caine-induced status epilepticus model [5]. Marimastat is also important for the treatment of advanced pancreatic cancer [6, 7, 8, 9]. Marimastat has been tested in over 400 patients in Phase I / II trials in various solid tumors. Most of these studies measured tumor-specific antigens (CEA for CRC, CA 19 / 9 for pancreatic cancer, CA 125 for ovarian cancer, and prostate-specific antigen for prostate cancer) as surrogate markers of biological activity.Meta-analysis of all these studies as well as analysis of individual studies showed that Marimastat treatment significantly reduced all four growth markers of tumor-specific antigens in a dose-dependent manner [9].

[0005] The beneficial effects of Marimastat (PKL-021, Fig. 1) described in the literature cited above promote more extensive research on the use of Marimastat in the treatment of numerous diseases and in-depth physicochemical research on Marimastat. A thorough literature analysis revealed that no information regarding the physicochemical properties of marimastat was provided, and reports on its synthesis were few (PL 174279 B1; Richard J Davenport, Robert J Watson: "An improved synthesis of the broad spectrum matrix metalloprotease inhibitor marimastat"; Tetrahedron Letters, Volume 41, Issue 41, 7.10.2000, pp. 7983-7986; Kai Jenssen, Katherina Sewald and Norbert Sewald: "Synthesis of Marimastat and a Marimastat Conjugate for Affinity Chromatography and Surface Plasmon Resonance Studies"; Bioconjugate Chem. 2004, 15, 594-600; Daniel E. Levy, France Lapierre, Weisheng Liang, Wenqing Ye, Christopher W. Lange, Xiaoyuan Li, Damian Grobelny, Marie Casabonne, David Tyrrell, Kevin Holme, Alex Nadzan and Richard E. Galardy: “Matrix Metalloproteinase Inhibitors: A Structure-Activity Study”; J. Med. Chem. 1998, 41, 199-223).The physicochemical properties of active substances used as drugs are crucial in the development of the synthesis process itself, play an important role in the manufacturing process of appropriate formulations, and finally influence the rates of release, absorption, and distribution within the body.

[0006] The development of synthesis processes for active substances possessing favorable physicochemical parameters, such as crystal size and its distribution, polymorphic forms and their stability, bulk density, and solubility, is crucial for the execution of filtration, washing, and drying processes and also influences the performance of pharmaceutical formulation processes. Throughout the entire technical process, the active substance can be converted into various polymorphic forms that are thermodynamically more stable but less soluble. Furthermore, phenomena such as the formation of so-called pseudopolymorphs—namely, solvates of the substance and solvent, and hydrates when the solvent incorporated into the crystal structure is water—may be observed.

[0007] When planning to obtain a high-quality active substance with a specific release profile and subsequently a pharmaceutical product, all of the above physicochemical properties of the substance must be taken into account. A properly characterized active substance will enable technicians to develop an optimal formulation production method and select appropriate equipment as well as the type of granulation.

[0008] The objective of the present invention is to produce crystalline polymorphic forms of marimastat and amorphous forms of marimastat, to determine their complete physicochemical properties, and to develop effective synthesis methods for them.

[0009] The subject of the present invention is a crystalline polymorphic form of marimastat, which is crystalline polymorphic form I, characterized by a powder X-ray diffraction pattern obtained by Cu Kα irradiation including main peaks at 2-theta° 6.11, 11.10, 12.19, 18.15, 19.57, 20.54, 20.74, 23.35, 24.50 and 26.20 ± 0.2.

[0010] Preferably, crystalline polymorphic form I is an anhydride.

[0011] Another subject of the present invention is a crystalline polymorphic form of marimastat that is crystalline polymorphic form II, which is characterized by a powder X-ray diffraction pattern obtained by Cu Kα irradiation including main peaks at 2-theta° 6.18, 12.26, 14.95, 19.74, 21.53, 24.55, 24.55, 27.73, 30.80 and 37.12 ± 0.2.

[0012] Preferably, crystalline polymorphic form II is anhydrous.

[0013] Another subject of the present invention is a crystalline polymorphic form of marimastat that is crystalline polymorphic form III, which is characterized by a powder X-ray diffraction pattern obtained by Cu Kα irradiation including main peaks at 2-theta° 6.32, 11.05, 12.60, 17.30, 18.36, 20.43, 20.93, 24.05, 26.83 and 40.30 ± 0.2.

[0014] Preferably, crystalline polymorphic form III is anhydrous.

[0015] Another object of the present invention is a mixture of crystalline polymorphic form I of marimastat and crystalline polymorphic form III of marimastat, which is characterized by a powder X-ray diffraction pattern obtained by Cu Kα irradiation including main peaks at 2-theta° 6.14, 6.30, 11.09, 12.22, 12.60, 18.43, 19.62, 20.51, 20.78 and 20.98 ± 0.2.

[0016] Another subject of the present invention is the amorphous form of marimastat.

[0017] Another object of the present invention is a method for preparing crystalline polymorphic form I of marimastat according to the present invention, wherein the method comprises the step of macerating an amorphous form of marimastat in a solvent selected from the group comprising 2-methyltetrahydrofuran, isopropyl ether, methyl tert-butyl ether, methylene chloride, ethanol, cyclohexane, 1,2-dichloroethane, trichloroethylene, isopropyl acetate, 2-butanone, pentane, and acetonitrile.

[0018] Preferably, the method comprises a) a step of maceration of the amorphous form of marimastat in a solvent while stirring at room temperature for 2 to 4 days; and b) a step of isolating the formed crystals, wherein the solvent is selected from the group comprising 2-methyltetrahydrofuran, isopropyl ether, methyl tert-butyl ether, methylene chloride, ethanol, cyclohexane, 1,2-dichloroethane, trichloroethylene, isopropyl acetate, 2-butanone, pentane, and acetonitrile.

[0019] Another object of the present invention is a method for preparing crystalline polymorphic form I of marimastat according to the present invention, wherein the method comprises the steps of preparing a saturated solution of marimastat in dimethyl sulfoxide and precipitating crystals using acetonitrile, ethyl acetate, or acetone.

[0020] Preferably, the method comprises a) preparing a saturated solution of marimastat in dimethyl sulfoxide at a temperature of 40 to 90°C; b) optionally filtering the solution and washing it with dimethyl sulfoxide; c) adding acetonitrile, ethyl acetate, or acetone to the solution to precipitate crystals and stirring the formed suspension at room temperature for 1 to 72 hours; and d) isolating the formed crystals.

[0021] Another object of the present invention is a method for producing crystalline polymorphic form I of marimastat according to the present invention, wherein the method comprises crystallization from ethanol.

[0022] Preferably, the method comprises a) a step of preparing a saturated solution of marimastat in 96% v / v ethanol or anhydrous ethanol; b) a step of stirring at room temperature for 5 to 7 days; and c) a step of isolating the formed crystals.

[0023] Another object of the present invention is a method for producing a crystalline polymorphic form II of marimastat according to the present invention, wherein the method comprises the step of macerating an amorphous form of marimastat in a solvent selected from chloroform or diethyl ether.

[0024] Preferably, the method comprises a) a step of maceration of the amorphous form of marimastat in a solvent while stirring at room temperature for 2 to 5 days; and b) a step of isolating the formed crystals, wherein the solvent is selected from chloroform or diethyl ether.

[0025] Another object of the present invention is a method for preparing crystalline polymorphic form II of marimastat according to the present invention, the method being characterized by comprising: a) preparing a saturated solution of marimastat in boiling methanol; b) adding 2-methyltetrahydrofuran, 1,2-dimethoxyethane, or chloroform to the solution; c) cooling the solution to a temperature of 30 to 40°C; and d) isolating the formed crystals.

[0026] Another object of the present invention is a method for preparing crystalline polymorphic form II of marimastat according to the present invention, the method being characterized by comprising: a) preparing a saturated solution of marimastat in methanol at a temperature of 55 to 65°C; b) mixing the solution with ethyl acetate at a temperature of 55 to 65°C; c) cooling the mixture to room temperature; and d) isolating the formed crystals.

[0027] Another object of the present invention is a method for preparing crystalline polymorphic form II of marimastat according to the present invention, the method comprising: a) preparing a saturated solution of marimastat in dimethyl sulfoxide at a temperature of 40 to 90°C; b) adding chloroform, methylene chloride, 2-methyltetrahydrofuran, or 1,2-dimethoxyethane to the solution; c) cooling the solution to a temperature of room temperature to 40°C; and d) isolating the formed crystals.

[0028] Another object of the present invention is a method for preparing crystalline polymorphic form II of marimastat according to the present invention, the method comprising: a) preparing a saturated solution of marimastat in dimethyl sulfoxide at a temperature of 50 to 65°C; b) mixing the solution with ethyl acetate at a temperature of 55 to 65°C; c) cooling the mixture to room temperature; and d) isolating the formed crystals.

[0029] Another object of the present invention is a method for preparing crystalline polymorphic form II of marimastat according to the present invention, the method being characterized by comprising: a) reacting (R)-N-((S)-3,3-dimethyl-1-methylamino-1-oxobutane-2-yl)-2-((S)-2,2-dimethyl-5-oxo-1,3-dioxolan-4-yl)-4-methylpentanamide with a 50% aqueous solution of hydroxylamine in a mixture of 2-methyltetrahydrofuran or dimethyl sulfoxide and chloroform at room temperature to boiling point; b) removing water by performing azeotropic distillation after the reaction is completed; and c) isolating the formed crystals.

[0030] Another object of the present invention is a method for preparing a crystalline polymorphic form III of marimastat according to the present invention, wherein the method comprises the steps of preparing a saturated solution of marimastat in methanol and precipitating crystals using ethyl acetate.

[0031] Preferably, the method comprises a) preparing a saturated solution of marimastat in boiling methanol; b) cooling the solution to 50 to 60°C; c) adding ethyl acetate to precipitate crystals; d) stirring the suspension while boiling for 20 to 60 minutes and azeotropically distilling the methanol; e) cooling the suspension to room temperature and stirring for 15 to 24 hours; and f) isolating the formed crystals.

[0032] Another object of the present invention is a method for preparing a crystalline polymorphic form III of marimastat according to the present invention, the method being characterized by comprising the steps of preparing a saturated solution of marimastat in dimethyl sulfoxide and precipitating crystals using a methanol solution in water.

[0033] Preferably, the method comprises a) preparing a saturated solution of marimastat in dimethyl sulfoxide at a temperature of 60 to 75°C; b) optionally filtering the solution; c) mixing the solution with a methanol solution in water at a ratio of 1:7 v / v; d) letting the solution stand until crystals precipitate; and e) isolating the formed crystals.

[0034] Another object of the present invention is a method for preparing a mixture of crystalline polymorphic form I of marimastat and crystalline polymorphic form III of marimastat according to the present invention, wherein the method comprises the step of macerating an amorphous form of marimastat in a solvent selected from the group comprising tetrahydrofuran, 1,2-dimethoxyethane, 2-propanol, 1-propanol, isoamyl alcohol, acetone, and heptane.

[0035] Preferably, the method comprises a) a step of maceration of the amorphous form of marimastat in a solvent while stirring at room temperature for 2 to 5 days; and b) a step of isolating the formed crystals, wherein the solvent is selected from the group comprising tetrahydrofuran, 1,2-dimethoxyethane, 2-propanol, 1-propanol, isoamyl alcohol, acetone, and heptane.

[0036] Another object of the present invention is a method for preparing a mixture of crystalline polymorphic form I of marimastat and crystalline polymorphic form III of marimastat according to the present invention, wherein the method comprises the steps of preparing a saturated solution of marimastat in methanol and precipitating crystals using acetonitrile, acetone, isopropyl acetate, or 2-methyltetrahydrofuran.

[0037] Preferably, the method comprises a) preparing a saturated solution of marimastat in methanol at a temperature of 60 to 65°C; b) mixing the solution with acetonitrile, isopropyl acetate, or boiling acetone; c) stirring the formed suspension at room temperature for 15 to 24 hours; and d) isolating the formed crystals.

[0038] Preferably, the method comprises a) preparing a saturated solution of marimastat in boiling methanol; b) cooling the solution to 50 to 60°C; c) adding 2-methyltetrahydrofuran to precipitate crystals; d) stirring the formed suspension while boiling for 20 to 60 minutes and azeotropically distilling the methanol; e) cooling the suspension to room temperature and stirring for 15 to 24 hours; and f) isolating the formed crystals.

[0039] Another object of the present invention is a method for preparing a mixture of crystalline polymorphic form I of marimastat and crystalline polymorphic form III of marimastat according to the present invention, wherein the method comprises the step of crystallizing in a boiling solvent selected from ethanol, n-propanol, or isopropanol.

[0040] Preferably, the method comprises a) preparing a saturated solution of marimastat in a boiling solvent selected from ethanol, n-propanol, or isopropanol; b) cooling the solution to room temperature and stirring for 15 to 24 hours; and c) isolating the formed crystals.

[0041] Another object of the present invention is a method for preparing a mixture of crystalline polymorphic form I of marimastat and crystalline polymorphic form III of marimastat according to the present invention, wherein the method comprises the steps of preparing a saturated solution of marimastat in boiling ethanol and precipitating crystals using ethyl acetate.

[0042] Preferably, the method comprises a) preparing a saturated solution of marimastat in boiling ethanol; b) adding ethyl acetate to precipitate crystals; c) stirring the formed suspension at room temperature for 15 to 24 hours; and d) isolating the formed crystals.

[0043] Another object of the present invention is a method for preparing a mixture of crystalline polymorphic form I of marimastat and crystalline polymorphic form III of marimastat according to the present invention, wherein the method is characterized by comprising the steps of preparing a saturated solution of marimastat in dimethylformamide and precipitating crystals using isopropyl acetate.

[0044] Preferably, the method comprises a) preparing a saturated solution of marimastat in dimethylformamide at a temperature of 70 to 85°C; b) adding isopropyl acetate to precipitate crystals; c) stirring the formed suspension at room temperature for 15 to 24 hours; and d) isolating the formed crystals.

[0045] Another object of the present invention is a pharmaceutical composition, characterized by comprising crystalline polymorphic forms I, II, or III of marimastat according to the present invention, a mixture of crystalline polymorphic forms I and III of marimastat according to the present invention, an amorphous form of marimastat according to the present invention or a mixture thereof, and a pharmaceutically acceptable carrier.

[0046] The pharmaceutical composition according to the present invention is administered by a route suitable for the type of drug form: intestinal (e.g., oral, sublingual, rectal), parenteral (e.g., intravenous, arterial, intramuscular, subcutaneous, by inhalation), or topically (e.g., intraocular, epidermal, vaginal).

[0047] The pharmaceutical composition according to the present invention may be provided in various pharmaceutical forms well known to a person skilled in the art, for example, known in the literature [Remington's Pharmaceutical Sciences, ed. 18, Mack Publ.Co.].

[0048] Preferably, the pharmaceutical composition is in the form of a tablet, modified-release tablet, pill, capsule, powder, granule, pellet, suspension, emulsion, solution, oral liquid form, e.g., syrup, injectable solution or suspension, infusion solution, eye drops, ointment, gel, suppository, globule, or therapeutic system, e.g., implant, vaginal ring, nanofiber.

[0049] Pharmaceutical forms for oral administration include, for example, tablets, pills, powders, granules, pellets, or capsules comprising a solid pharmaceutically acceptable carrier such as corn starch, lactose, sucrose, sorbitol, talc, stearic acid, magnesium stearate, dicalcium phosphate, or gum. Tablets or granules may be coated or processed to provide a dosage unit that offers the benefit of sustained action.

[0050] Various materials, including various polymeric acids and mixtures of polymeric acids and materials such as shellac, ethyl alcohol, or cellulose acetate, can be used to form these protective layers or coating layers.

[0051] Pharmaceutical forms for injection and infusion include sterile aqueous solutions, aqueous-organic solutions and non-aqueous solutions, suspensions, dry materials, and solutions or tablets for implantation. To prepare suspensions, excipients such as polysorbates, lecithin, copolymers of polyoxyethylene and polyoxypropylene, peptizers such as phosphates, polyphosphates, and citrates, water-soluble polymers such as carboxymethylcellulose, methylcellulose, and polyvinylpyrrolidone, and gums or gelatin are used to ensure that the medicinal substance is uniformly dispersed in the liquid phase. Injectable formulations may include pharmaceutically acceptable excipients such as pH adjusters and buffers, isotonic agents, and preservatives. Dry materials can be prepared as solutions or suspensions immediately by diluting with a suitable solvent ( ex tempore It is intended to manufacture.

[0052] Another object of the present invention is a pharmaceutical composition according to the present invention for use as a medicine.

[0053] Another object of the present invention is a pharmaceutical composition according to the present invention for use in the prevention and treatment of diseases associated with the overactivity of extracellular matrix metalloproteinases selected from the group including epilepsy after stroke, epilepsy after trauma, epilepsy after brain surgery, hypoxic-ischemic encephalopathy, malignant neoplasms, vascular malformations, amyotrophic lateral sclerosis, multiple sclerosis, snake venom poisoning, endometriosis, hemorrhoids, arthritis, neurological diseases, circulatory diseases, diabetes, and diabetic complications selected from the group including diabetic retinopathy and diabetic foot.

[0054] Another subject of the present invention is a crystalline polymorphic form I, II, or III of marimastat according to the present invention for use as a pharmaceutical agent, a mixture of crystalline polymorphic form I of marimastat according to the present invention and crystalline polymorphic form III of marimastat according to the present invention, an amorphous form of marimastat according to the present invention, or a mixture thereof.

[0055] Another subject of the present invention is a crystalline polymorphic form I, II, or III of Marimarstat according to the present invention for use in the prevention and treatment of diseases associated with the overactivity of extracellular matrix metalloproteinases selected from the group including epilepsy after stroke, epilepsy after trauma, epilepsy after brain surgery, hypoxic-ischemic encephalopathy, malignant neoplasm, vascular malformation, amyotrophic lateral sclerosis, multiple sclerosis, snake venom poisoning, endometriosis, hemorrhoids, arthritis, neurological diseases, circulatory diseases, diabetes, and diabetic complications selected from the group including diabetic retinopathy and diabetic foot, a mixture of crystalline polymorphic form I of Marimarstat according to the present invention and crystalline polymorphic form III of Marimarstat according to the present invention, an amorphous form of Marimarstat according to the present invention, or a mixture thereof.

[0056] The selection of drug dosage and administration regimen depends on the type of disease, the patient's age, weight, and health, and may be determined by an expert based on known therapeutic regimens. In the treatment of epilepsy after stroke, epilepsy after trauma, epilepsy after brain surgery, hypoxic-ischemic encephalopathy, malignant neoplasms, vascular malformations, amyotrophic lateral sclerosis, multiple sclerosis, snake venom poisoning, endometriosis, hemorrhoids, arthritis, neurological diseases, circulatory diseases, diabetes, and diabetic complications such as diabetic retinopathy and diabetic foot, the appropriate dose of the crystalline polymorphic form of Marimastat according to the present invention, a mixture of crystalline polymorphic form I and crystalline polymorphic form III of Marimastat according to the present invention, the amorphous form of Marimastat according to the present invention, or a mixture thereof may be 0.05 to 50 mg / kg per day, preferably 0.1 to 10 mg / kg per day. An appropriate dose may be administered to the patient once or several times a day, either alone or in combination with other medicinal substances. These substances may be administered simultaneously or sequentially in the form of a single preparation or separate preparations, or in an order and at time intervals determined by a specialist. Brief explanation of the drawing

[0057] The subject of the present invention is described in more detail in the embodiments and drawings. Figure 1 shows the structural formula of Marimarstat (PKL-021). Figure 2 shows the XRPD pattern for the amorphous form. Figure 3 shows the XRPD pattern for polymorphic form I. Figure 4 shows the XRPD pattern for polymorphic form II. Figure 5 shows the XRPD pattern for polymorphic form III. Figure 6 shows the XRPD pattern for a mixture of polymorphic forms I and III. Figure 7 shows a thermogram from TG analysis for polymorphic form I, where TGA is indicated by the top line, heat flow by the middle line, and first derivative by the bottom line. Figure 8 shows a thermogram from TG analysis for polymorphic form II, where TGA is indicated by the top line, heat flow by the middle line, and first derivative by the bottom line. Figure 9 shows a thermogram from TG analysis for polymorphic form III, where TGA is indicated by the top line, heat flow by the middle line, and first derivative by the bottom line. Figure 10 shows a photograph of polymorphic form I from an optical microscope (polarized, x40 magnification). Figure 11 shows a photograph of polymorphic form II from an optical microscope (x400 magnification). Figure 12A shows a photograph of polymorphic form III from an electron microscope. Figure 12B shows a photograph of polymorphic form III from an optical microscope (x100 magnification). Figure 13 shows the solid state of polymorphic form I. 13This shows the C CPMAS NMR spectrum. Fig. 14 shows the solid state of polymorphic form II. 13 This shows the C CPMAS NMR spectrum. Fig. 15 shows the solid state of polymorphic form III. 13 This shows the C CPMAS NMR spectrum. Figure 16 shows the solid state of a mixture of polymorphic forms I and III. 13 This shows the C CPMAS NMR spectrum. Specific details for implementing the invention

[0058] The obtained polymorphic forms of marimastat were characterized by X-ray powder diffraction (XRPD). Tables 1, 2, 3, and 4 contain the positions and intensities of the diffraction lines of the obtained forms I, II, III, and mixtures of forms I and III. Characteristic reflections used to identify the three polymorphic forms and mixtures of forms I and III, as well as to determine the degree of their crystalline purity, are indicated in bold in Tables 1, 2, 3, and 4.

[0059] Characteristic reflections of form I occur at 2-theta° angles of 6.11, 11.10, 12.19, 18.15, 19.57, 20.54, 20.74, 23.35, 24.50, and 26.20 ± 0.2.

[0060] For type II, characteristic reflections occur at 2-theta° angles of 6.18, 12.26, 14.95, 19.74, 21.53, 24.55, 24.55, 27.73, 30.80, and 37.12 ± 0.2.

[0061] For form III, characteristic reflections occur at 2-theta° angles of 6.32, 11.05, 12.60, 17.30, 18.36, 20.43, 20.93, 24.05, 26.83, and 40.30 ± 0.2.

[0062] The mixture of types I and III has characteristic reflections at 2-theta° angles of 6.14, 6.30, 11.09, 12.22, 12.60, 18.43, 19.62, 20.51, 20.78, and 20.98 ± 0.2.

[0063]

[0064]

[0065]

[0066]

[0067]

[0068]

[0069]

[0070]

[0071] Example 1

[0072] Manufacturing in amorphous form

[0073] 2 g of marimastat was dissolved in water (500 ml) at room temperature. The clear solution was frozen in an acetone / dry ice bath and freeze-dried for 48 hours. An amorphous material was obtained in quantitative yield.

[0074] Example 2

[0075] Manufacturing in amorphous form

[0076] 50 mg of marimastat was ground in a ball mill at 30 Hz for 1.5 to 3 hours. An amorphous material was obtained in quantitative yield.

[0077] Example 3

[0078] Preparation of Crystalline Polymorphic Form I

[0079] The amorphous form was suspended in a solvent selected from the group consisting of 2-methyltetrahydrofuran (2-MeTHF), isopropyl ether, methyl tert-butyl ether (MTBE), methylene chloride, ethanol (EtOH), cyclohexane, 1,2-dichloroethane, trichloroethylene, isopropyl acetate, 2-butanone, pentane, and acetonitrile, and then stirred at room temperature for 2 to 4 days. The white suspension was placed in an Eppendorfer. Polymorphic form I was centrifuged and dried at room temperature under high vacuum.

[0080] Example 4

[0081] Preparation of Crystalline Polymorphic Form I

[0082] Marimastert was dissolved in dimethyl sulfoxide (DMSO) at a temperature of 50°C, filtered through a Schott filter, and washed with DMSO. Then, acetonitrile was added to the clear solution, and the resulting suspension was stirred at room temperature for 3 days. The crystals were filtered, washed with acetonitrile, and dried at room temperature under high vacuum to obtain Form I with a yield of 70%.

[0083] Example 5

[0084] Preparation of Crystalline Polymorphic Form I

[0085] Marimastert was heated in anhydrous ethanol or 96% v / v ethanol until dissolved, and then stirred at room temperature for 7 days. The crystals were filtered, washed with anhydrous ethanol or 96% v / v ethanol, and dried at room temperature and under high vacuum to obtain Form I with yields of 75% and 65%, respectively.

[0086] Example 6

[0087] Preparation of Crystalline Polymorphic Form I

[0088] Marimastert was dissolved in DMSO at a temperature of 50°C. Then, ethyl acetate was added to the clear solution, and the resulting suspension was stirred at room temperature for 2 hours. The crystals were filtered, washed with ethyl acetate, and dried at room temperature under high vacuum to obtain Form I with a yield of 60%.

[0089] Example 7

[0090] Preparation of Crystalline Polymorphic Form I

[0091] Marimastert was dissolved in DMSO at a temperature of 50°C. Then, acetone was added to the clear solution, and the resulting suspension was stirred at room temperature for 2 hours. The crystals were filtered, washed with acetone, and dried at room temperature under high vacuum to obtain Form I with a yield of 46%.

[0092] Example 8

[0093] Preparation of Crystalline Polymorphic Form II

[0094] The amorphous form was suspended in chloroform or diethyl ether and stirred at room temperature for 2 to 4 days. The white suspension was placed in an Eppendorfer. Polymorphic form II was centrifuged and dried at room temperature under high vacuum.

[0095] Example 9

[0096] Preparation of Crystalline Polymorphic Form II

[0097] Marimastert was dissolved in boiling methanol (MeOH), and 2-methyltetrahydrofuran (2-MeTHF) was added. After cooling to room temperature, the formed suspension was stirred for 24 hours. The crystals were filtered, washed with 2-MeTHF, and dried at room temperature under high vacuum to obtain Form II with a yield of 60%.

[0098] Example 10

[0099] Preparation of Crystalline Polymorphic Form II

[0100] Marimastert was dissolved in boiling methanol (MeOH), and 2-methyltetrahydrofuran (2-MeTHF) was added. After cooling to a temperature of 40°C, the crystals were filtered, washed with 2-MeTHF, and dried at room temperature under high vacuum to obtain Form II with a yield of 48%.

[0101] Example 11

[0102] Preparation of Crystalline Polymorphic Form II

[0103] Marimastert was dissolved in boiling methanol (MeOH), and 1,2-dimethoxyethane was added. After cooling to 40°C, the crystals were filtered, washed with 1,2-dimethoxyethane, and dried at room temperature under high vacuum to obtain Form II with a yield of 61%.

[0104] Example 12

[0105] Preparation of Crystalline Polymorphic Form II

[0106] Marimastert was dissolved in boiling methanol (MeOH), and chloroform was added. After cooling to 40°C, the crystals were filtered, washed with chloroform, and dried at room temperature under high vacuum to obtain Form II with a yield of 40%.

[0107] Example 13

[0108] Preparation of Crystalline Polymorphic Form II

[0109] Marimastert was dissolved in dimethyl sulfoxide (DMSO) at a temperature of 50°C, and chloroform was added. After cooling to 40°C, the crystals were filtered, washed with chloroform, and dried at room temperature under high vacuum to obtain Form II with a yield of 89%.

[0110] Example 14

[0111] Preparation of Crystalline Polymorphic Form II

[0112] Marimastert was dissolved in methanol (MeOH) at 60°C, and the solution was added to ethyl acetate (AcOEt) at 60°C. After cooling to room temperature, the crystals were filtered, washed with AcOEt, and dried at room temperature under high vacuum to obtain Form II with a yield of 83%.

[0113] Example 15

[0114] Preparation of Crystalline Polymorphic Form II

[0115] Marimastert was dissolved in dimethyl sulfoxide (DMSO) at 60°C, and the solution was added to ethyl acetate (AcOEt) at 60°C. After cooling to room temperature, the crystals were filtered, washed with AcOEt, and dried at room temperature under high vacuum to obtain Form II with a yield of 55%.

[0116] Example 16

[0117] Preparation of Crystalline Polymorphic Form II

[0118] Marimastert was dissolved in dimethyl sulfoxide (DMSO) at a temperature of 50°C, and methylene chloride was added. After cooling to room temperature, the crystals were filtered, washed with methylene chloride, and dried at room temperature under high vacuum to obtain Form II with a yield of 90%.

[0119] Example 17

[0120] Preparation of Crystalline Polymorphic Form II

[0121] 2-methyltetrahydrofuran (2-MeTHF, 900 ml) was poured into (R)-N-((S)-3,3-dimethyl-1-methylamino-1-oxobutane-2-yl)-2-((S)-2,2-dimethyl-5-oxo-1,3-dioxolane-4-yl)-4-methylpentanamide (45 g) (obtained by the method disclosed in Tetrahedron Letters

[12] ) and heated to a temperature of 40 to 45°C to obtain a clear solution. Then, a 50% aqueous solution of hydroxylamine (50% NH2OH, 29 ml) was added, and the reaction was carried out at a temperature of 40 to 45°C for 1 to 2 hours. After the reaction was completed, acetone (90 ml) was added, and the reaction mixture was stirred for 20 to 30 minutes while boiling. Subsequently, water was removed by azeotropic distillation, and once distillation was complete, the suspension was stirred at room temperature for 15 to 20 hours. The crystals were filtered, washed with 2-MeTHF, and dried at room temperature to obtain Form II with a yield of 88% (37 g).

[0122] Example 18

[0123] Preparation of Crystalline Polymorphic Form II

[0124] (R)-N-((S)-3,3-dimethyl-1-methylamino-1-oxobutane-2-yl)-2-((S)-2,2-dimethyl-5-oxo-1,3-dioxolane-4-yl)-4-methylpentanamide (10 g, 28 mmol, 1 equivalent) (obtained by the method disclosed in Tetrahedron Letters

[12] ) was dissolved in 14 mL of dimethyl sulfoxide (DMSO) and 14 mL of chloroform at room temperature. Then, NH2OH (5.2 mL, 50% aqueous solution, 84 mmol, 3.0 equivalents) was added. The suspension was stirred at room temperature for 1 hour (TLC monitoring: 10% CH3OH / CH2Cl2). Then, 9 mL of acetone was added dropwise. The reaction mixture was stirred for 30 minutes while boiling. Then, water was removed from the reaction mixture by azeotropic distillation from a mixture of DMSO, chloroform, and acetone. The volume of the distilled solvent was replenished by successively adding additional amounts of chloroform. Then, the entire volume of chloroform was distilled off, and 185 ml of chloroform was added dropwise while maintaining boiling to obtain a clear solution of marimastat in DMSO. Then, the reaction mixture was cooled to room temperature and stirred for 17 hours. The crystals were filtered, washed with chloroform (15 ml) and acetone (3 × 15 ml), and dried to obtain Form II with an 83% yield (7.71 g).

[0125] Example 19

[0126] Preparation of Crystalline Polymorphic Form III

[0127] A Marimastert suspension (202 g) was added to boiling methanol (1.2 L) and stirred until dissolved. Then, the clear solution was cooled to 60°C, and ethyl acetate (2 L) was added. The suspension was stirred while boiling for 20 minutes, and then the methanol was removed by azeotropic distillation. The suspension was cooled to room temperature and stirred for 24 hours. The crystals were filtered, washed with ethyl acetate, and dried at room temperature to obtain 192 g of Form III with a yield of 95%.

[0128] Example 20

[0129] Preparation of Crystalline Polymorphic Form III

[0130] Marimastert was dissolved in DMSO at a temperature of 72°C. The solution was filtered through a Schott filter and washed with DMSO. The clear solution was added to an H2O / MeOH (7:1 v / v) mixture, and the resulting suspension was stirred at room temperature for 24 hours. The crystals were filtered, washed with H2O and MeOH, and dried at room temperature under high vacuum to obtain Form III with a yield of 60%.

[0131] Example 21

[0132] Preparation of a mixture of crystalline polymorphic forms I and III

[0133] The amorphous form was suspended in a solvent selected from the group consisting of tetrahydrofuran, 1,2-dimethoxyethane, 2-propanol, 1-propanol, isoamyl alcohol, acetone, and heptane, and then stirred at room temperature for 4 days. The white suspension was placed in an Eppendorfer. The mixture of forms I and III was centrifuged and dried at room temperature under high vacuum.

[0134] Example 22

[0135] Preparation of a mixture of crystalline polymorphic forms I and III

[0136] Marimastert was dissolved in MeOH at a temperature of 65°C, and the clear solution was added to acetonitrile at room temperature. The formed suspension was stirred at room temperature for 24 hours. The crystals were filtered, washed with acetonitrile, and dried at room temperature under high vacuum to obtain a mixture of forms I and III in a yield of 80%.

[0137] Example 23

[0138] Preparation of a mixture of crystalline polymorphic forms I and III

[0139] Marimastert was dissolved in MeOH at a temperature of 65°C, and the clear solution was added to boiling acetone. The formed suspension was stirred at room temperature for 24 hours. The crystals were filtered, washed with acetone, and dried at room temperature under high vacuum to obtain a mixture of forms I and III in a yield of 60%.

[0140] Example 24

[0141] Preparation of a mixture of crystalline polymorphic forms I and III

[0142] Marimastert was dissolved in MeOH under boiling conditions, and then isopropyl acetate was added. The resulting suspension was stirred at room temperature for 24 hours. The crystals were filtered, washed with isopropyl acetate, and dried at room temperature under high vacuum to obtain a mixture of forms I and III in a yield of 89%.

[0143] Example 25

[0144] Preparation of a mixture of crystalline polymorphic forms I and III

[0145] Marimastert was suspended in n-propanol and stirred until dissolved while boiling. The clear solution was cooled to room temperature and stirred for 24 hours. The crystals were filtered, washed with n-propanol, and dried at room temperature under high vacuum to obtain a mixture of forms I and III with a yield of 83%.

[0146] Example 26

[0147] Preparation of a mixture of crystalline polymorphic forms I and III

[0148] Marimastert was dissolved in boiling anhydrous ethanol (EtOH). The clear solution was cooled to room temperature and stirred for 24 hours. The crystals were filtered, washed with EtOH, and dried at room temperature under high vacuum to obtain a mixture of forms I and III in a yield of 67%.

[0149] Example 27

[0150] Preparation of a mixture of crystalline polymorphic forms I and III

[0151] Marimastert was suspended in isopropanol and stirred until dissolved while boiling. The clear solution was cooled to room temperature and stirred for 24 hours. The crystals were filtered, washed with isopropanol, and dried at room temperature under high vacuum to obtain a mixture of forms I and III in a yield of 70%.

[0152] Example 28

[0153] Preparation of a mixture of crystalline polymorphic forms I and III

[0154] Marimastert was dissolved in ethanol under boiling conditions, and then ethyl acetate was added. The resulting suspension was stirred at room temperature for 24 hours. The crystals were filtered, washed with ethyl acetate, and dried at room temperature under high vacuum to obtain a mixture of forms I and III with a yield of 77%.

[0155] Example 29

[0156] Preparation of a mixture of crystalline polymorphic forms I and III

[0157] Marimastert was dissolved in dimethylformamide (DMF) at a temperature of 85°C, and then isopropyl acetate was added dropwise. The formed suspension was stirred at room temperature for 24 hours. The crystals were filtered, washed with isopropyl acetate, and dried at room temperature under high vacuum to obtain a mixture of forms I and III in a yield of 95%.

[0158] Example 30

[0159] Preparation of a mixture of crystalline polymorphic forms I and III

[0160] The marimastat suspension was stirred in boiling methanol until dissolved. Then, the clear solution was cooled to 60°C, and 2-methyltetrahydrofuran (2-MeTHF) was added. The suspension was stirred while boiling for 20 minutes, and then the methanol was removed by azeotropic distillation. The suspension was cooled to room temperature and stirred for 24 hours. The crystals were filtered, washed with 2-MeTHF, and dried at room temperature under high vacuum to obtain a mixture of forms I and III in a yield of 98%.

[0161] Example 31

[0162] X-ray Powder Diffraction (XRPD) Analysis

[0163] Powder diffraction patterns were collected using a Rigaku MiniFlex600 instrument with radiation of Cu Kα1 wavelength and λ = 1.54 Å for crystalline polymorphic forms I, II, and mixtures of forms I and III, and using a Bruker D8 Advance instrument with radiation of Cu Kα1 wavelength and λ = 1.54 Å for crystalline polymorphic form III and amorphous forms. XRPD patterns for the amorphous form, crystalline polymorphic forms I, II, III, and mixtures of forms I and III are shown in Figs. 2, 3, 4, 5, and 6, respectively.

[0164] Example 32

[0165] Thermogravimetric Analysis (TGA)

[0166] Thermogravimetric analysis was performed on crystalline polymorphism forms I, II, and III using a Mettler Toledo TGA / DSC1 instrument. Samples were weighed into aluminum pans sealed with a perforated aluminum lid.

[0167] Crystalline polymorphic form I

[0168] TG analysis was performed at a rate of 10°C / min in the range of 25°C to 320°C. The TG profile showed a weight loss of 0.3% at temperatures from 25°C to 195°C. Decomposition occurred above 200°C. TG analysis confirms that crystalline polymorphic form I of marimastat exists in an anhydrous form. A thermogram from the TG analysis for crystalline polymorphic form I is shown in Fig. 7.

[0169] Crystalline Polymorphic Form II

[0170] TG analysis was performed at a rate of 10°C / min in the range of 25°C to 320°C. A weight loss of 0.2% was observed at temperatures from 25°C to 195°C. Decomposition occurred above 200°C. TG analysis confirms that crystalline polymorphic form II of marimastat exists in an anhydrous form. A thermogram from TG analysis for crystalline polymorphic form II is shown in Fig. 8.

[0171] Crystalline polymorphic form III

[0172] This sample was analyzed by TGA-EGA. The analysis was performed at a rate of 10°C / min at temperatures ranging from 25°C to 320°C. A weight loss of 0.4% was observed at temperatures ranging from 25°C to 195°C. Decomposition occurred above 200°C. TG analysis confirms that crystalline polymorphic form III of marimastat exists in an anhydrous form. A thermogram from TG analysis for crystalline polymorphic form III is shown in Fig. 9.

[0173] conclusion

[0174] Analysis of X-ray powder diffraction and thermogravimetric analysis results indicates that three crystalline polymorphic forms of pure anhydride marimastat and a mixture of crystalline polymorphic forms I and III were obtained and characterized.

[0175] Example 33

[0176] Stability study

[0177] Stability studies of the amorphous form, pure crystalline polymorphic forms I, II, and III, and a mixture of crystalline polymorphic forms I and III were conducted under the following conditions: 25°C / 100% RH (relative humidity) and 40°C / 75% RH (relative humidity) for 7 days, 30°C / 100 mbar for 1 hour, room temperature for 1 year, and also under increased temperature (above 150°C). XRPD analysis of the tested samples showed no changes in the diffraction line images, which proves that all three polymorphic forms of marimastat, the mixture of forms I and III, and the amorphous form are stable.

[0178] A stability study in various solvents was also performed on the crystalline polymorphic forms of marimastat obtained by stirring forms I, II, and III in a suitable solvent at room temperature for 4 days. XRPD analysis of the samples indicated that the most thermodynamically stable form is form I (Table 5).

[0179]

[0180] XRPD analysis of the samples showed that in six solvents, a mixture of the three polymorphic forms was converted to pure Form I (Items 1–6, Table 5), and in three solvents, it was converted to a mixture of Forms I and III (Items 14–16, Table 5). In the remaining seven solvents, a mixture of the initial three polymorphic forms was obtained. Therefore, when a mixture of all three forms was used for maceration in different solvents, it was observed that Forms II and III were completely lost, Form II was completely lost, or all three forms remained in the mixture. Complete loss of Form I was not observed in any experiment. The above experiments demonstrate that both Forms III and II are kinetic forms, and that Form I of Marimastert is the most thermodynamically stable form.

[0181] The stability of Form II in various solvents at room temperature was tested. Form II was suspended in 1 ml of solvent, and the resulting suspension was stirred at room temperature of 25°C for 4 days to conduct the experiment. Then, the precipitate was filtered, the sample was dried at 30°C / 100 mbar for 1 hour, and XRPD analysis was performed. The results obtained are presented in Table 6.

[0182]

[0183] XRPD analysis of the samples showed a complete conversion from Form II to Form I in two solvents, acetonitrile and methanol (Items 2 and 8, Table 6). In other solvents, Form II is the stable form. The formation of Form III was not observed. The above experiments confirmed that Form I of marimastat possesses the highest thermodynamic stability.

[0184] Example 34

[0185] Microscopic examination

[0186] Microscopic examinations were performed using an optical microscope or an electron microscope. Microscopic examinations of the three pure forms of marimastat, as well as mixtures of forms I and III, revealed that these forms possess different crystal morphologies (Figs. 10, 11, and 12) and different bulk densities, which is particularly important in the formulation manufacturing process. Generally, forms I and II have the highest and most comparable bulk densities.

[0187] Example 35

[0188] Solubility test

[0189] Solubility tests were performed by dissolving samples of the pure form of marimastat in water. The results of the solubility tests for crystalline polymorphic forms I, II, and III showed that form III dissolved the fastest in water, followed by form II, and form I dissolved the slowest.

[0190] Example 36

[0191] 13 C CPMAS NMR and 15 Analysis of N CPMAS NMR Spectra

[0192] All solid-state spectra were recorded on Bruker Avance III 600 MHz and Bruker Avance III 400 MHz spectrometers. Samples of polymorphic forms I, II, and III, as well as mixtures of polymorphic forms I and III, were placed in a 4 mm zirconium oxide rotor and rotated at the magic angle at 12 kHz (600 MHz) and 8 kHz (400 MHz). Using a Cross Polarization Magic Angle Spinning (CPMAS) pulse sequence 13 C and 15 N spectra were acquired. The spectral quality, defined by the Full Width at Half Height (FWHH), suggests that each tested sample is a regular and well-organized material with a high degree of crystallinity. No amorphous phase was observed in the tested materials. Based on the measurements performed, it was confirmed that three polymorphic forms exist in a pure form. NMR analysis of the mixture of forms I and III was also performed. Forms I, II, and III, and the mixture of forms I and III 13 The C CPMAS NMR spectra are shown in Figs. 13, 14, 15, and 16.

[0193] Polymorphic form I:

[0194] 13 C NMR (solid state) , d (C=O) = 175.7ppm, d (C=O) = 171.1ppm, δ (C=O) = 169.8ppm, d (C-H) = 73.1ppm, d (C-H) = 61.0ppm, d (C-H) = 50.3ppm, d (C-H) = 43.3ppm, d (Cquat) = 37.5ppm, d (CH2) = 29.6ppm, d (C-H3) = 27.4ppm, d (C-H) = 26.9ppm, d (C-H3) = 26.5ppm, d (C-H3) = 24.9ppm, d (C-H3) = 23.9ppm.

[0195] 15 N NMR (solid state) , δ = 168.4ppm, δ = 117.2ppm, δ = 116.5ppm.

[0196] Polymorphic Form II:

[0197] 13 C NMR (solid state) , d (C=O) = 176.0ppm, d (C=O) = 175.3ppm, d (C=O) = 171.1ppm, δ (C=O) = 169.7ppm, d (C-H) = 73.9ppm, d (C-H) = 72.3ppm, d (C-H) = 61.0ppm, d (C-H) = 50.7ppm, d (C-H) = 49.2ppm, d (C-H) = 43.3ppm, d (C-H2) = 37.6ppm, d (Cquat) = 37.4ppm, d (CH2) = 30.6ppm, d (C-H3) = 27.2ppm, δ (C-H) = 26.6ppm, d (C-H3) = 25.5ppm, d (C-H3) = 25.0ppm, d(C-H3) = 23.6ppm.

[0198] 15 N NMR (solid state) , δ = 168.3ppm, δ = 117.1ppm, δ = 116.5ppm.

[0199] Polymorphic Form III:

[0200] 13 C NMR (solid state) , d (C=O) = 174.6ppm, d (C=O) = 171.4ppm, d (C=O) = 169.6ppm, d (C-H) = 69.2ppm, d (C-H) = 60.6ppm, d (C-H) = 48.6ppm, d (C-H) = 41.0ppm, d (C-quat) = 38.0ppm, d (C-H2) = 27.8ppm, d (C-H3) = 27.8ppm, d (C-H) = 26.3ppm, d (C-H3) = 25.6ppm, d (C-H3) = 25.4ppm, d (C-H3) = 24.9ppm.

[0201] 15 N NMR (solid state) , δ = 166.6ppm, δ = 117.1ppm, δ = 116.2ppm.

[0202] Mixture of polymorphic forms I and III:

[0203] 13 C NMR (solid state) , d (C=O) = 175.7ppm, d (C=O) = 174.6ppm, d (C=O) = 171.4ppm, δ (C=O) = 171.1ppm, δ (C=O) = 169.7ppm, d (C-H) = 73.0ppm, d (C-H) = 69.1ppm, d (C-H) = 60.7ppm, d (C-H) = 50.2ppm, d (Cquat)= 48.7ppm, δ (C-H) = 43.2ppm, δ (C-H) = 41.0ppm, δ (Cquat) = 37.9ppm, δ (Cquat) = 37.5ppm, δ (CH2) = 29.2ppm, δ (C-H3) = 27.6ppm, δ (C-H3) = 27.3ppm, δ (C-H) = 26.4ppm, δ (C-H) = 26.3ppm, δ (C-H3) = 25.5ppm, δ (C-H3) = 25.3ppm, δ (C-H3) = 24.8ppm, δ (C-H3) = 23.9ppm.

[0204] 15 N NMR (solid state) , δ = 168.4ppm, δ = 166.6ppm, δ = 117.2ppm, δ = 116.4ppm.

[0205] Example 37

[0206] Preparation of tablets containing a mixture of crystalline polymorphic forms I and III

[0207] Tablets containing a mixture of crystalline polymorphic forms I and III were prepared by the wet granulation method according to the following steps:

[0208] 1. Preparation of granulated solution

[0209] 2. Granulation

[0210] 3. Granule size calibration before drying

[0211] 4. Drying

[0212] 5. Adjustment of granule particle size after drying

[0213] 6. Mixing of granules and external components

[0214] 7. Tableting

[0215] Step 1: Preparation of granulated solution

[0216] The binder was added to water and stirred until completely dissolved.

[0217] Step 2: Granulation

[0218] The mixture of internal phase components was granulated in a high-speed granulator with an aqueous solution of the binder obtained in Step 1.

[0219] Step 3: Adjusting Granule Particle Size Before Drying

[0220] Before the drying process, the wet granules were adjusted in particle size using a conical mill or a vibrating granulator. The wet granules can also be adjusted in particle size manually using a steel sieve with an appropriate mesh size.

[0221] Step 4: Drying

[0222] The granules were dried using a fluidized bed dryer or a tray dryer at a target temperature of approximately 55°C (40 to 65°C) and dried until a required weight loss rate (0.2 to 3.0%) was reached.

[0223] Step 5: Adjustment of granule particle size after drying

[0224] The particle size of the dried granules was adjusted manually using a steel sieve, a conical mill, or a vibrating granulator.

[0225] Step 6: Mixing of granules and external components

[0226] All external components (excluding the lubricant) were mixed with the granules in a low-speed mixer, and the speed was adjusted according to the size of the mixer tank and its filling amount. The lubricant was added at the end of the mixing step.

[0227] Step 7: Tableting

[0228] Tableting was performed for each dosage of tablets containing a mixture of crystalline polymorphic forms I and III (the dosage is obtained proportionally by compressing to match a specific target tablet weight).

[0229] Method for preparing tablets containing a mixture of crystalline polymorphic forms I and III

[0230] Concentration of a mixture of crystalline polymorphic forms I and III in the product: 10 to 40%.

[0231] Drug dosage: Obtained proportionally through the target tablet weight.

[0232]

[0233] References:

[0234]

Claims

Claim 1 A crystalline polymorphic form of marimastat, wherein the crystalline polymorphic form is crystalline polymorphic form I, characterized by a powder X-ray diffraction pattern obtained by Cu Kα irradiation including main peaks at 2-theta° 6.11, 11.10, 12.19, 18.15, 19.57, 20.54, 20.74, 23.35, 24.50, and 26.20 ± 0.

2. Claim 2 A crystalline polymorphic form of marimastat, characterized as being anhydrous in claim 1. Claim 3 A crystalline polymorphic form of marimastat, wherein the crystalline polymorphic form is crystalline polymorphic form II, characterized by a powder X-ray diffraction pattern obtained by Cu Kα irradiation including main peaks at 2-theta° 6.18, 12.26, 14.95, 19.74, 21.53, 24.55, 24.55, 27.73, 30.80, and 37.12 ± 0.

2. Claim 4 A crystalline polymorphic form of Marimarstat, characterized in that, in paragraph 3, the crystalline polymorphic form II is an anhydrous. Claim 5 A crystalline polymorphic form of marimastat, wherein the crystalline polymorphic form is crystalline polymorphic form III, characterized by a powder X-ray diffraction pattern obtained by Cu Kα irradiation including main peaks at 2-theta° 6.32, 11.05, 12.60, 17.30, 18.36, 20.43, 20.93, 24.05, 26.83 and 40.30 ± 0.

2. Claim 6 A crystalline polymorphic form of marimastat characterized as being anhydrous in paragraph 5. Claim 7 A mixture of crystalline polymorphic form I of marimastat and crystalline polymorphic form III of marimastat, characterized by a powder X-ray diffraction pattern obtained by Cu Kα irradiation including main peaks at 2-theta° 6.14, 6.30, 11.09, 12.22, 12.60, 18.43, 19.62, 20.51, 20.78 and 20.98 ± 0.

2. Claim 8 Amorphous form of Marimastat. Claim 9 A method for preparing crystalline polymorphic form I of marimastat as defined in claim 1 or 2, comprising the step of macerating the amorphous form of marimastat in a solvent for 2 to 4 days, wherein the solvent is selected from the group comprising 2-methyltetrahydrofuran, isopropyl ether, methyl tert-butyl ether, methylene chloride, ethanol, cyclohexane, 1,2-dichloroethane, trichloroethylene, isopropyl acetate, 2-butanone, pentane, and acetonitrile. Claim 10 A method for preparing crystalline polymorphic form I of marimastat as defined in claim 1 or 2, characterized by comprising the steps of preparing a saturated solution of marimastat in dimethyl sulfoxide and precipitating crystals using acetonitrile, ethyl acetate, or acetone. Claim 11 A method for preparing crystalline polymorphic form I of marimastat as defined in claim 1 or 2, characterized by including a step of crystallizing from ethanol. Claim 12 A method for preparing a crystalline polymorphic form II of marimastat as defined in claim 3 or 4, characterized by comprising the step of macerating the amorphous form of marimastat in a solvent selected from chloroform or diethyl ether. Claim 13 A method for preparing crystalline polymorphic form II of marimastat as defined in claim 3 or 4, comprising: a) preparing a saturated solution of marimastat in boiling methanol; b) adding 2-methyltetrahydrofuran, 1,2-dimethoxyethane, or chloroform to the solution; c) cooling the solution to a temperature of 30 to 40°C; and d) isolating the formed crystals. Claim 14 A method for preparing crystalline polymorphic form II of marimastat as defined in claim 3 or 4, comprising: a) preparing a saturated solution of marimastat in methanol at a temperature of 55 to 65°C; b) mixing the solution with ethyl acetate at a temperature of 55 to 65°C; c) cooling the mixture to room temperature; and d) isolating the formed crystals. Claim 15 A method for preparing a crystalline polymorphic form II of marimastat as defined in claim 3 or 4, comprising: a) preparing a saturated solution of marimastat in dimethyl sulfoxide at a temperature of 40 to 90°C; b) adding chloroform, methylene chloride, 2-methyltetrahydrofuran, or 1,2-dimethoxyethane to the solution; c) cooling the solution to a temperature of room temperature to 40°C; and d) isolating the formed crystals. Claim 16 A method for preparing a crystalline polymorphic form II of marimastat as defined in claim 3 or 4, comprising: a) preparing a saturated solution of marimastat in dimethyl sulfoxide at a temperature of 50 to 65°C; b) mixing the solution with ethyl acetate at a temperature of 55 to 65°C; c) cooling the mixture to room temperature; and d) isolating the formed crystals. Claim 17 A method for preparing a crystalline polymorphic form II of marimastat as defined in claim 3 or 4, comprising: a) reacting (R)-N-((S)-3,3-dimethyl-1-methylamino-1-oxobutane-2-yl)-2-((S)-2,2-dimethyl-5-oxo-1,3-dioxolan-4-yl)-4-methylpentanamide with a 50% aqueous solution of hydroxylamine in a mixture of 2-methyltetrahydrofuran or dimethyl sulfoxide and chloroform at room temperature to boiling point; b) removing water by performing azeotropic distillation after the reaction is completed; and c) isolating the formed crystals. Claim 18 A method for preparing crystalline polymorphic form III of marimastat as defined in claim 5 or 6, characterized by comprising the steps of preparing a saturated solution of marimastat in methanol and precipitating crystals using ethyl acetate. Claim 19 A method for preparing crystalline polymorphic form III of marimastat as defined in claim 5 or 6, characterized by comprising the steps of preparing a saturated solution of marimastat in dimethyl sulfoxide and precipitating crystals using a methanol solution in water. Claim 20 A method for preparing a mixture of crystalline polymorphic form I of marimastat and crystalline polymorphic form III of marimastat as defined in claim 7, comprising the step of macerating the amorphous form of marimastat in a solvent for 2 to 5 days, wherein the solvent is selected from the group comprising tetrahydrofuran, 1,2-dimethoxyethane, 2-propanol, 1-propanol, isoamyl alcohol, acetone, and heptane. Claim 21 A method for preparing a mixture of crystalline polymorphic form I of marimastat and crystalline polymorphic form III of marimastat as defined in claim 7, characterized by comprising the steps of preparing a saturated solution of marimastat in methanol and precipitating crystals using acetonitrile, acetone, isopropyl acetate, or 2-methyltetrahydrofuran. Claim 22 A method for preparing a mixture of crystalline polymorphic form I of marimastat and crystalline polymorphic form III of marimastat as defined in claim 7, characterized by comprising the step of crystallizing in a boiling solvent selected from ethanol, n-propanol, or isopropanol. Claim 23 A method for preparing a mixture of crystalline polymorphic form I of marimastat and crystalline polymorphic form III of marimastat as defined in claim 7, characterized by comprising the steps of preparing a saturated solution of marimastat in boiling ethanol and precipitating crystals using ethyl acetate. Claim 24 A method for preparing a mixture of crystalline polymorphic form I of marimastat and crystalline polymorphic form III of marimastat as defined in claim 7, characterized by comprising the steps of preparing a saturated solution of marimastat in dimethylformamide and precipitating crystals using isopropyl acetate. Claim 25 A pharmaceutical composition characterized by comprising a crystalline polymorphic form of marimastat defined in any one of claims 1 to 6, a mixture of crystalline polymorphic form I of marimastat defined in claim 7 and crystalline polymorphic form III of marimastat, an amorphous form of marimastat defined in claim 8 or a mixture thereof, and a pharmaceutically acceptable carrier. Claim 26 A pharmaceutical composition according to claim 25, characterized in that it is in the form of a tablet, modified-release tablet, pill, capsule, powder, granule, pellet, suspension, emulsion, solution, oral liquid form, e.g., syrup, injectable solution or suspension, infusion solution, eye drops, ointment, gel, suppository, globule, or therapeutic system, e.g., implant, vaginal ring, nanofiber. Claim 27 A pharmaceutical composition for use as a medicine, according to paragraph 25 or 26. Claim 28 A pharmaceutical composition for use in the prevention and treatment of diseases associated with the hyperactivity of extracellular matrix metalloproteinases selected from the group including epilepsy after stroke, epilepsy after trauma, epilepsy after brain surgery, hypoxic-ischemic encephalopathy, malignant neoplasm, vascular malformation, amyotrophic lateral sclerosis, multiple sclerosis, snake venom poisoning, endometriosis, hemorrhoids, arthritis, neurological diseases, circulatory diseases, diabetes mellitus, and diabetic complications selected from the group including diabetic retinopathy and diabetic foot. Claim 29 A crystalline polymorphic form of marimastat defined in any one of claims 1 to 6 for use as a pharmaceutical agent, a mixture of crystalline polymorphic form I of marimastat defined in claim 7 and crystalline polymorphic form III of marimastat, an amorphous form of marimastat defined in claim 8, or a mixture thereof. Claim 30 A crystalline polymorphic form of Marimastat defined in any one of claims 1 to 6, a mixture of crystalline polymorphic form I of Marimastat defined in claim 7 and crystalline polymorphic form III of Marimastat defined in claim 8, or an amorphous form of Marimastat defined therein, for use in the prevention and treatment of diseases associated with the hyperactivity of extracellular matrix metalloproteinases selected from the group including epilepsy after stroke, epilepsy after trauma, epilepsy after brain surgery, hypoxic-ischemic encephalopathy, malignant neoplasm, vascular malformation, amyotrophic lateral sclerosis, multiple sclerosis, snake venom poisoning, endometriosis, hemorrhoids, arthritis, neurological diseases, circulatory diseases, diabetes, and diabetic complications selected from the group including diabetic retinopathy and diabetic foot.

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