Rosaxostat-piperazine eutectic crystal as well as preparation method and application thereof

Abstract

The invention provides a rosaxostat-piperazine eutectic crystal as well as a preparation method and application thereof. The rosaxostat-piperazine eutectic crystal is formed by combining rosaxostat and piperazine according to a molar ratio of 1: 1. As a novel crystal form, the rosaxostat-piperazine eutectic crystal provided by the invention can improve the dissolution rate of rosaxostat without changing the covalent bond of the rosaxostat, thereby enhancing the drug effect and bioavailability. The preparation method adopts a grinding reaction crystallization method to prepare the rosaxostat-piperazine eutectic crystal, and is simple in process, mild in condition, good in repeatability, green, environment-friendly and suitable for large-scale production.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of medicine, and relates to a roxadustat-piperazine co-crystal and a preparation method and application thereof. BACKGROUND

[0002] Chronic kidney disease (CKD) has posed a great threat to global public health, and there are about 120 million CKD patients in China, and the incidence rate in the adult population is as high as 10.8%. Anemia is a common complication of CKD patients, and the incidence rate increases with the decline of renal function. According to research, the anemia prevalence rate of CKD dialysis patients in China is as high as 98.2%, and the anemia prevalence rate of non-dialysis patients is 52.1%. At present, the dominant drug for treating CKD anemia is erythropoiesis stimulating agent (ESA), but the application of ESA is limited by serious adverse events such as cardiovascular events. In recent years, domestic and foreign researchers have begun to explore a new type of erythropoiesis stimulating agent, i.e., hypoxia-inducible factor prolyl hydroxylase (HIF-PHI) inhibitor.

[0003] Roxadustat is the first HIF-PHI drug for treating renal anemia in the world, and has been approved for marketing in multiple countries. Roxadustat is one of the substrates of prolyl hydroxylase (PH), which affects the role of PH enzyme in maintaining the balance of HIF generation and degradation, thereby endogenously correcting anemia. Roxadustat, chemically named N-[(4-hydroxy-1-methyl-7-phenoxy-3-isoquinoline) carbonyl] glycine, was first marketed in China by the original research company FibroGen on December 17, 2018, and the trade name is Auryon. Its structural formula is as shown below:

[0004]

[0005] Roxadustat has poor water solubility, and the poor physicochemical properties will seriously affect its bioavailability and thus affect the therapeutic effect. Many researchers have tried to find a method for improving the solubility of roxadustat through various free acid crystal forms, salts and solubility-related researches of roxadustat. For example, patents WO2019042641 and WO2019030711 disclose roxadustat-L-proline co-crystals, CN111320583A discloses a DL-proline co-crystal of roxadustat, but since the proline structure does not have ultraviolet absorption, it is not suitable for content determination by conventional liquid phase method. Therefore, CN114057643A discloses a roxadustat-L-tryptophan co-crystal.

[0006] WO2019030711 also discloses a co-crystal of Roxadustat with nicotinamide, urea, CN110218184A discloses a co-crystal of Roxadustat amide compound, including Roxadustat-cinnamamide co-crystal, Roxadustat-benzamide co-crystal, Roxadustat-nicotinamide co-crystal. IN201741007950A discloses a co-crystal form of Roxadustat with caffeine, adenine, thymine, saccharin. Although the above has improved the solubility of Roxadustat to a certain extent, more solutions are still needed to supplement the content of Roxadustat multi-component crystals.

[0007] It is an object of the present application to provide an improved solid state form of Roxadustat, in particular a co-crystal of Roxadustat, which is physically stable against temperature stress. It is a further object of the present application to provide an improved solid state form of Roxadustat, in particular a co-crystal of Roxadustat, which is chemically stable against photodegradation, essentially free of organic solvents, which is characterized by improved dissolution / solubility and / or which is characterized by improved powder properties, such as flowability, bulk density and compressibility.

[0008] Therefore, it is of great significance to develop a new crystal form of Roxadustat to improve the stability, solubility, flowability and compressibility of Roxadustat. SUMMARY

[0009] The present application aims to provide a Roxadustat-piperazine co-crystal and a preparation method and application thereof. As a new crystal form, the Roxadustat-piperazine co-crystal can improve the stability, solubility and flowability of Roxadustat without changing the covalent bond of Roxadustat, thereby improving the drug efficacy and bioavailability. The Roxadustat-piperazine co-crystal is prepared by a grinding reaction crystallization method, which is simple, mild, reproducible, green and environmentally friendly, and suitable for large-scale production.

[0010] One of the objects of the present application is to provide a Roxadustat-piperazine co-crystal, which is formed by combining Roxadustat and piperazine at a molar ratio of 1:1.

[0011] Preferably, the roxadustat-piperazine co-crystal has an X-ray powder diffraction pattern, expressed in terms of diffraction angles 2θ, using Cu-kα radiation, with characteristic peaks at 3.50 ± 0.2°, 7.19 ± 0.2°, 10.18 ± 0.2°, 10.88 ± 0.2°, 11.76 ± 0.2°, 13.84 ± 0.2°, 14.36 ± 0.2°, 17.09 ± 0.2°, 18.68 ± 0.2°, 20.64 ± 0.2°, 21.86 ± 0.2°, 22.88 ± 0.2°, 23.32 ± 0.2°, 24.56 ± 0.2°, 26.57 ± 0.2°, 27.91 ± 0.2°, 29.77 ± 0.2°, 32.07 ± 0.2°, 33.64 ± 0.2°.

[0012] Preferably, the roxadustat-piperazine co-crystal has an X-ray powder diffraction pattern, expressed in terms of diffraction angles 2θ, using Cu-kα radiation, with characteristic peaks at 3.50 ± 0.2°, 7.19 ± 0.2°, 10.18 ± 0.2°, 10.88 ± 0.2°, 11.76 ± 0.2°, 13.84 ± 0.2°, 14.36 ± 0.2°, 17.09 ± 0.2°, 18.68 ± 0.2°, 20.64 ± 0.2°, 21.86 ± 0.2°, 22.88 ± 0.2°, 23.32 ± 0.2°, 24.56 ± 0.2°, 26.57 ± 0.2°, 27.91 ± 0.2°, 29.77 ± 0.2°, 32.07 ± 0.2°, 33.64 ± 0.2°.

[0013] The DSC pattern of the roxadustat-piperazine co-crystal has a characteristic endothermic peak at 213℃ ± 2℃;

[0014] Preferably, the TG pattern of the roxadustat-piperazine co-crystal starts losing weight at 110℃, decomposes while dissolving, and completely decomposes after 600℃.

[0015] The second object of the present application is to provide a preparation method of the roxadustat-piperazine co-crystal according to the first object, characterized in that the preparation method comprises:

[0016] The roxadustat-piperazine co-crystal is obtained by dissolving roxadustat and piperazine hexahydrate in a good solvent, stirring for 10-30 min.

[0017] Preferably, the good solvent comprises any one or a combination of at least two of acetone, dichloromethane, 1,4-dioxane, butanone, N-methyl pyrrolidone, acetonitrile, ethylene glycol methyl ether, DMF, DMSO, tetrahydrofuran, methanol, n-propanol or isopropanol.

[0018] Preferably, the molar ratio of the addition of roxadustat and piperazine hexahydrate is 1:2-2:1.

[0019] Preferably, the temperature of the grinding is 10-40℃, and the time of the grinding is 10-30min.

[0020] Preferably, the concentration of the Roxadustat in the good solvent is 5-100mg / mL.

[0021] Preferably, the preparation method further comprises solid-liquid separation and drying of the mixture obtained after stirring for 10-30min.

[0022] The third object of the present application is to provide a use of the Roxadustat-piperazine co-crystal according to any one of the objects in the preparation of a HIF modulating drug.

[0023] Compared with the prior art, the present application has the following beneficial effects:

[0024] The Roxadustat-piperazine co-crystal prepared by the grinding reaction crystallization method of the present application has the advantages of simple process, mild conditions, good repeatability, green environmental protection, and suitability for large-scale production.

[0025] The Roxadustat-piperazine co-crystal prepared by the present application, as a new crystal form, can improve the stability, solubility and flowability of Roxadustat without changing the covalent bond of Roxadustat, thereby improving the drug efficacy and bioavailability. BRIEF DESCRIPTION OF DRAWINGS

[0026] Figure 1 The powder X-ray diffraction (PXRD) pattern of the Roxadustat-piperazine co-crystal prepared in Example 1.

[0027] Figure 2 The differential scanning calorimetry (DSC) pattern of the Roxadustat-piperazine co-crystal prepared in Example 1.

[0028] Figure 3 The Fourier transform infrared spectroscopy (FT-IR) pattern of the Roxadustat-piperazine co-crystal prepared in Example 1. DETAILED DESCRIPTION

[0029] It should be noted that the following detailed description is exemplary in nature and is intended to provide further description of the present application. 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 the present application belongs. The technical solutions of the present application are further illustrated below through specific embodiments. Those skilled in the art should understand that the embodiments are only to help understand the present application and should not be regarded as specific limitations of the present application.

[0030] All alternatives, modifications, and equivalents of the methods and materials disclosed herein can be made within the scope of the invention, which is defined by the following claims. One skilled in the art can recognize, or be able to ascertain using no more than routine experimentation, many equivalents for the specific embodiments described herein. The invention is not intended to be limited to the embodiments described herein, but is to be accorded the full scope that the claims afford to the invention. In the event that any one or more of the incorporated literature, patents, and similar materials differs from or contradicts this application, including but not limited to defined terms, term application, described techniques, and the like, this application controls.

[0031] It should be further recognized that certain of the inventors' features, while described in the context of a number of separate embodiments, can also be provided in combination in a single embodiment. Conversely, various features of the inventors' invention, while described in the context of a single embodiment, can also be provided separately or in any appropriate subcombination.

[0032] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as those commonly understood by one of ordinary skill in the art to which this invention belongs. All patents and publications referred to in this specification are incorporated herein by reference in their entirety.

[0033] Unless defined otherwise, the following definitions shall apply for the purposes of this application. For the purposes of this application, the chemical elements are identified in accordance with the Periodic Table of the Elements, CAS version, and the elements are identified by their atomic number as specified in the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 75thEd., inside cover, and specific gravity is as defined therein. Unless otherwise indicated, all chemicals used in the preparation and practice of this invention were obtained from commercial suppliers such as Sigma-Aldrich, Acros, Fisher Scientific, VWR, and Alfa Aesar unless otherwise noted. The practice of this invention is not limited to the materials, reagents, conditions, or the like, set forth in the following examples.

[0034] The terms "comprising" or "including" or "having" are intended to be open-ended terms that include both the recited elements and other elements. The terms "consisting of and "consisting essentially of are intended to be closed terms that do not include other elements.

[0035] The term "co-crystal" or "co-crystalline" refers to a composition of two or more phases; phase refers to something that is identical in composition, crystal structure, and properties.

[0036] The term "XRD pattern" or "PXRD" refers to an X-ray powder diffraction pattern.

[0037] The term "substantially as shown" means that a substantially pure "crystal form" has at least 50%, or at least 60%, or at least 70%, or at least 80%, or at least 90%, or at least 95%, or at least 99% of its peaks appearing in the given X-ray powder diffraction pattern. As the content of a certain crystal form in a sample gradually decreases, some diffraction peaks attributable to that crystal form in its X-ray powder diffraction pattern may decrease due to factors such as the instrument's detection sensitivity.

[0038] The term "relative intensity" refers to the ratio of the intensity of the other peaks to the intensity of the first strongest peak in a set of diffraction peaks belonging to a certain crystal form, when the intensity of the first strongest peak is defined as 100%.

[0039] In the context of this invention, the 2θ (also known as 2theta or diffraction peak) values ​​in X-ray powder diffraction patterns are expressed in degrees (°).

[0040] When referring to spectra and / or data in figures, the term "diffraction peak" refers to a feature that a person skilled in the art would not attribute to background noise.

[0041] The measurement of the 2θ or diffraction peaks in the X-ray powder diffraction pattern of the crystal is subject to experimental error. The measurement of the 2θ or diffraction peaks in the X-ray powder diffraction pattern may vary slightly between different machines and between different samples. The experimental error or difference may be + / - 0.2 units, + / - 0.1 units, or + / - 0.05 units. Therefore, the value of the 2θ or diffraction peaks cannot be considered absolute.

[0042] The differential scanning calorimetry (DSC) curve of the crystal has experimental errors. The position and peak value of the endothermic peak may vary slightly between one machine and another, and between one sample and another. The experimental error or difference may be less than or equal to 5°C, or less than or equal to 4°C, or less than or equal to 3°C, or less than or equal to 2°C, or less than or equal to 1°C. Therefore, the peak position or peak value of the DSC endothermic peak cannot be regarded as absolute.

[0043] The thermogravimetric analysis (TGA) curves of the crystals are subject to experimental error. The endothermic curves or weight loss rates may vary slightly between different machines and between different samples. The experimental error or difference may be less than or equal to 0.004%, 0.003%, 0.002%, or 0.001%. Therefore, the thermogravimetric analysis curves or their weight loss rates cannot be considered absolute.

[0044] In the context of this invention, all figures disclosed herein are approximate values, regardless of whether the words "approximately" or "about" are used. The value of each figure may vary by 1%, 2%, or 5%, etc. When "approximately" is used to describe the 2θ (also known as 2theta or diffraction peak) value of an X-ray powder diffraction peak, "approximately" means that the 2θ value may vary by + / - 0.2 units, + / - 0.1 units, or + / - 0.05 units.

[0045] Specific test results:

[0046] The following analytical methods and equipment were used to characterize the roxadustat-piperazine cocrystal of the present invention:

[0047] (1) Instruments for XRPD testing: X-ray powder diffractometer;

[0048] Instrument model: Rigaku D / max-2500 (Japan);

[0049] Test method: Cu target Ka, voltage 40KV, current 100mA, test angle 2-40°, step size 8° / min, exposure time 0.2s, light tube slit width 1mm, detector slit width 2.7mm.

[0050] (2) DSC testing instrument: differential calorimeter

[0051] Instrument Model: Mettler Toledo DSC1 / 500

[0052] Test method: heating rate 10℃ / min, nitrogen flow rate of protective gas 50mL / min.

[0053] (3) Instruments for TGA testing: thermogravimetric analyzer;

[0054] Instrument model: Mettler Toledo TGA / DSC1 (Switzerland);

[0055] Test method: The atmosphere is nitrogen, and the heating rate is 10℃ / minute;

[0056] (4) Instrument used for in vitro dissolution rate testing: liquid chromatography;

[0057] Instrument model: Waters 2695 series high performance liquid chromatography system;

[0058] Test method: The absorbance of the solution was measured at the maximum wavelength of 300 nm. The concentration of the drug in the solution was calculated by measuring the RST concentration from 2 μg / mL to 300 μg / mL using a standard curve. 2 =0.9999.

[0059] The specific implementation method is as follows:

[0060] Example 1

[0061] Weigh 50 mg of piperazine hexahydrate and dissolve it in 7 mL of mixed solvent (dichloromethane:methanol volume ratio of 1:1) at room temperature. Add 90 mg of roxadustat, suspend and stir for 15 min, filter and dry to obtain the product.

[0062] The samples obtained above were analyzed using X-ray powder diffraction. The results are shown in the figure. Figure 1 ,from Figure 1 It can be seen that the X-ray powder diffraction pattern of the roxadustat-piperazine eutectic, expressed as a diffraction angle 2θ, has characteristic peaks at 3.50, 6.68, 7.19, 10.18, 10.88, 11.76, 13.10, 13.68, 13.84, 14.36, 14.62, 16.5, 17.09, 18.36, 18.68, 20.01, 20.64, 21.44, 21.86, 22.88, 23.32, 24.56, 25.31, 26.57, 26.94, 27.91, 28.7, 29.15, 29.77, 31.27, 32.07, 33.64, 38.38, and 39.12.

[0063] Differential scanning calorimetry (DSC) was performed on the samples obtained above, and the results are shown in the figure. Figure 2 ,from Figure 2 It can be seen that the DSC spectrum of dolutegravir-m-hydroxybenzoic acid cocrystal has a characteristic endothermic peak at 214℃.

[0064] Thermogravimetric analysis of the samples obtained above revealed that they began to lose weight at approximately 110°C, subsequently melting and decomposing simultaneously, and completely decomposing after 600°C.

[0065] An experiment was designed for the samples obtained above according to the current Chinese Pharmacopoeia guidelines for solubility testing. The solubility of the samples in a phosphate buffer solution with pH = 6.8 at 37℃ was measured, and the results are shown in Table 1. The results indicate that the solubility of the roxadustat-piperazine cocrystal is 2.8 times that of roxadustat.

[0066] The angle of repose of the sample obtained above was measured, and the angle of repose of the product was 44°.

[0067] Table 1

[0068] Substance Solubility (pg / mL) Roxadustat 295.70 Roxadustat-piperazine co-crystal 824.26

[0069] Example 2

[0070] Weigh 50 mg of piperazine hexahydrate, dissolve it in 14 mL of acetone at room temperature, add 90 mg of roxadustat, suspend and stir for 15 min, filter and dry to obtain the product.

[0071] The same characterization method was used on Example 2 as on Example 1, and it can be seen that the crystal form obtained is the same as that of Example 1, and the XRD and TG results are also the same as those of Example 1.

[0072] The product obtained in Example 2 was subjected to the same tests as in Example 1, and the same characterization results were obtained, thus demonstrating the robustness of the process.

[0073] The product obtained in Example 2 was subjected to the same solubility and angle of repose tests as in Example 1. The results showed that the solubility of the roxadustat-piperazine eutectic was 3 times that of roxadustat, and the angle of repose of the roxadustat-piperazine eutectic was 40°.

[0074] Example 3

[0075] Weigh 100 mg of piperazine hexahydrate, dissolve it in 7 mL of acetone at room temperature, add 90 mg of roxadustat, suspend and stir for 15 min, filter and dry to obtain the product.

[0076] The same characterization method was used on Example 3 as on Example 1. It can be seen that the crystal form obtained is the same as that of Example 1, and the XRD and TG results are also the same as those of Example 1.

[0077] The product obtained in Example 3 was subjected to the same solubility and angle of repose tests as in Example 1. The results showed that the solubility of the roxadustat-piperazine eutectic was 2.8 times that of roxadustat, and the angle of repose of the roxadustat-piperazine eutectic was 41°.

[0078] Example 4

[0079] Weigh 25 mg of piperazine hexahydrate, dissolve it in 18 mL of dichloromethane at room temperature, add 90 mg of roxadustat, suspend and stir for 15 min, filter and dry to obtain the product.

[0080] The same characterization method was used on Example 4 as on Example 1. It can be seen that the crystal form obtained is the same as that of Example 1, and the XRD and TG results are also the same as those of Example 1.

[0081] The product obtained in Example 4 was subjected to the same solubility and angle of repose tests as in Example 1. The results showed that the solubility of the roxadustat-piperazine eutectic was 3.1 times that of roxadustat, and the angle of repose of the roxadustat-piperazine eutectic was 40°.

[0082] Example 5

[0083] Weigh 100 mg of piperazine hexahydrate, dissolve it in 9 mL of DMSO at room temperature, add 90 mg of roxadustat, suspend and stir for 15 min, filter and dry to obtain the product.

[0084] The same characterization method was used on Example 5 as on Example 1. It can be seen that the crystal form obtained is the same as that of Example 1, and the XRD and TG results are also the same as those of Example 1.

[0085] The product obtained in Example 5 was subjected to the same solubility and angle of repose tests as in Example 1. The results showed that the solubility of the roxadustat-piperazine eutectic was 2.6 times that of roxadustat, and the angle of repose of the roxadustat-piperazine eutectic was 38°.

[0086] Example 6

[0087] Weigh 50 mg of piperazine hexahydrate, dissolve it in 14 mL of methanol at room temperature, add 90 mg of roxadustat, suspend and stir for 15 min, filter and dry to obtain the product.

[0088] The same characterization method was used on Example 6 as on Example 1. It can be seen that the crystal form obtained is the same as that of Example 1, and the XRD and TG results are also the same as those of Example 1.

[0089] The product obtained in Example 6 was subjected to the same solubility and angle of repose tests as in Example 1. The results showed that the solubility of the roxadustat-piperazine eutectic was 2.7 times that of roxadustat, and the angle of repose of the roxadustat-piperazine eutectic was 42°.

[0090] The applicant declares that the above description is only a specific embodiment of the present invention, but the protection scope of the present invention is not limited thereto. Those skilled in the art should understand that any changes or substitutions that can be easily conceived by those skilled in the art within the technical scope disclosed in the present invention fall within the protection and disclosure scope of the present invention.

Claims

1. A roxadustat-piperazine co-crystal, characterized by, The roxadustat-piperazine co-crystal is combined from roxadustat and piperazine in a molar ratio of 1:

1.

2. The roxadustat-piperazine co-crystal of claim 1, characterized in that, The X-ray powder diffraction pattern of the roxadustat-piperazine co-crystal, expressed in terms of diffraction angle 2θ, has characteristic peaks at 3.50±0.2°, 7.19±0.2°, 10.18±0.2°, 10.88±0.2°, 11.76±0.2°, 13.84±0.2°, 14.36±0.2°, 17.09±0.2°, 18.68±0.2°, 20.64±0.2°, 21.86±0.2°, 22.88±0.2°, 23.32±0.2°, 24.56±0.2°, 26.57±0.2°, 27.91±0.2°, 29.77±0.2°, 32.07±0.2°, 33.64±0.2°, using Cu-kα radiation.

3. The roxadustat-piperazine co-crystal of claim 2, wherein, The X-ray powder diffraction pattern of the roxadustat-piperazine co-crystal, expressed in terms of diffraction angle 2θ, has characteristic peaks at 6.68±0.2°, 13.10±0.2°, 13.68±0.2°, 14.62±0.2°, 16.50±0.2°, 18.36±0.2°, 20.01±0.2°, 21.44±0.2°, 25.31±0.2°, 26.94±0.2°, 28.70±0.2°, 29.15±0.2°, 31.27±0.2°, 38.38±0.2°, 39.12±0.2°, using Cu-kα radiation.

4. The roxadustat-piperazine co-crystal of claim 1, wherein, The DSC pattern of the roxadustat-piperazine co-crystal has a characteristic endothermic peak at 213℃±2℃; Preferably, the TG pattern of the roxadustat-piperazine co-crystal starts losing weight at 110℃, decomposes while dissolving, and completely decomposes after 600℃.

5. A process for preparing roxadustat-piperazine co-crystal according to any one of claims 1-4, characterized in that, The preparation method comprises: dissolving roxadustat and piperazine hexahydrate in a good solvent, stirring for 10-30 min to obtain the roxadustat-piperazine co-crystal.

6. The production method according to claim 5, wherein The good solvent comprises any one or a combination of at least two of acetone, dichloromethane, 1,4-dioxane, butanone, N-methyl pyrrolidone, acetonitrile, ethylene glycol methyl ether, DMF, DMSO, tetrahydrofuran, methanol, n-propanol or isopropanol.

7. The preparation method according to claim 5, characterized in that, The addition molar ratio of the roxadustat and piperazine hexahydrate is 1:2-2:

1.

8. The preparation method according to claim 5, characterized in that, The concentration of the roxadustat in the good solvent is 5-100 mg / mL.

9. The preparation method according to claim 5, characterized in that, The preparation method further comprises solid-liquid separation and drying of the mixture obtained after stirring for 10-30 min.

10. Use of the roxadustat-piperazine co-crystal according to any one of claims 1-4 in the preparation of a HIF modulating drug.

Images