Crystal of nitroimidazole compound and preparation method and application thereof

Abstract

The invention relates to a crystal of a nitroimidazole compound as well as a preparation method and application thereof, in particular to a crystal I, a crystal II and a crystal III of S-ornidazole disodium phosphate, and further relates to application of the crystals in preparation of anti-anaerobe infection drugs and anti-protozoan drugs.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical crystal forms, specifically relating to various crystals and uses of a nitroimidazole compound, and more specifically, to crystals I, II, and III of levonornidazole phosphate disodium, their preparation methods, and uses. Background Technology

[0002] Levo-ornidazole phosphate disodium is a prodrug of levonidazole, a potent anti-anaerobic and antiprotozoal drug. It is also a newly developed third-generation nitroimidazole derivative with higher efficacy, shorter treatment duration, better tolerability, and wider distribution in the body than metronidazole.

[0003] Patent CN101177433A mentions a levonornidazole disodium phosphate pentahydrate and its preparation method, but lacks data characterization; patent CN102731571A mentions a levonornidazole disodium phosphate hexahydrate and its preparation method; and patent CN109748934A mentions a levonornidazole disodium phosphate heptahydrate.

[0004] Because levonornidazole phosphate disodium is unstable under high temperature and high humidity, it is urgent to develop a more stable crystal form of levonornidazole phosphate disodium to meet clinical needs in order to improve the stability of the drug. Summary of the Invention

[0005] A first aspect of the present invention provides crystal I of levonornidazole phosphate disodium pentahydrate, wherein the levonornidazole phosphate disodium pentahydrate has the following structure:

[0006]

[0007] The X-ray powder diffraction pattern of crystal I includes a range of approximately 6.1 ± 0.2. o 15.1±0.2 o 18.3±0.2 o 23.9±0.2 o 24.3±0.2 o and 26.6±0.2 o The diffraction peak at the 2θ angle.

[0008] A second aspect of the present invention provides crystal II of levonornidazole phosphate disodium hexahydrate, wherein the levonornidazole phosphate disodium hexahydrate has the following structure:

[0009]

[0010] The X-ray powder diffraction pattern of crystal II includes approximately 6.0 ± 0.2 o 15.0±0.2 o15.7±0.2 o 16.9±0.2 o 24.0±0.2 o and 27.0±0.2 o The diffraction peak at the 2θ angle.

[0011] A third aspect of the present invention provides crystal III of levonornidazole phosphate disodium hexahydrate, wherein the levonornidazole phosphate disodium hexahydrate has the following structure:

[0012]

[0013] The X-ray powder diffraction pattern of crystal III includes approximately 6.0 ± 0.2 o 11.7±0.2 o 12.1±0.2 o 15.7±0.2 o 19.6±0.2 o and 31.9±0.2 o The diffraction peak at the 2θ angle.

[0014] A fourth aspect of the present invention provides a pharmaceutical composition comprising i) any one or more of the crystals of the present invention: crystal I of levonornidazole phosphate disodium pentahydrate, crystal II of levonornidazole phosphate disodium hexahydrate, and crystal III of levonornidazole phosphate disodium hexahydrate; ii) one or more pharmaceutically acceptable carriers.

[0015] A fourth aspect of the invention provides the use of crystal I, crystal II, crystal III, or any one or more of the pharmaceutical compositions of the invention in the preparation of anti-anaerobic bacterial infection drugs and anti-protozoal infection drugs.

[0016] The fifth aspect of the present invention provides a method for preparing crystal I of levonornidazole phosphate disodium pentahydrate, the method comprising the following steps:

[0017] Disodium levonornidazole phosphate and solvent I are mixed, heated to 40-50℃ to dissolve, filtered, and the filtrate is cooled to crystallize, thus obtaining crystal I;

[0018] The solvent I is selected from water, ethanol, isopropanol, acetone or acetonitrile.

[0019] A sixth aspect of the present invention provides a method for preparing crystal II of levonornidazole phosphate disodium hexahydrate, the method comprising the following steps:

[0020] Disodium levonornidazole phosphate and solvent II are mixed, heated to 40-50℃ to dissolve, filtered, and the filtrate is cooled to crystallize, thus obtaining crystal II;

[0021] The solvent II is selected from a mixture of at least two of water, ethanol, propanol, or acetone.

[0022] The seventh aspect of the present invention provides a method for preparing crystals III of levonornidazole phosphate disodium hexahydrate, the method comprising the following steps:

[0023] Disodium levonornidazole phosphate and solvent III are mixed, heated to 40-50℃ to dissolve, filtered, the filtrate is cooled to 30-35℃, then cooled to crystallize, filtered, and the filter cake is washed with acetone to obtain crystal III;

[0024] The solvent III is selected from a mixture of at least two of water, ethanol, propanol, or acetone. Attached Figure Description

[0025] Figure 1 XRPD pattern of crystal I of levonornidazole phosphate disodium pentahydrate.

[0026] Figure 2 XRPD pattern of crystal II of levonornidazole phosphate disodium hexahydrate.

[0027] Figure 3 XRPD pattern of crystal III of levonornidazole phosphate disodium hexahydrate.

[0028] Figure 4 This is a schematic diagram of the smallest asymmetric structural unit in the unit cell of crystal I of levonornidazole phosphate disodium pentahydrate.

[0029] Figure 5 This is a schematic diagram of the smallest asymmetric structural unit in the unit cell of crystal II of levonornidazole phosphate disodium hexahydrate.

[0030] Figure 6 This is a schematic diagram of the smallest asymmetric structural unit in the unit cell of crystal III of levonornidazole phosphate disodium hexahydrate.

[0031] Figure 7 Microscopic image of crystal I of levonornidazole phosphate disodium pentahydrate.

[0032] Figure 8 Microscopic image of crystal II of levonornidazole phosphate disodium hexahydrate.

[0033] Figure 9 Microscopic image of crystal III of levonornidazole phosphate disodium hexahydrate. Detailed Implementation

[0034] definition

[0035] Unless otherwise defined below, all technical and scientific terms used herein are intended to have the same meaning as commonly understood by one of ordinary skill in the art. References to technical terms herein refer to techniques commonly understood in the art, including variations or equivalent substitutions of techniques that are obvious to one of ordinary skill in the art. While it is believed that the following terms will be well understood by one of ordinary skill in the art, the following definitions are set forth to better explain the invention.

[0036] The terms “including,” “comprising,” “having,” “containing,” or “involving,” as used herein, and their other variations thereof, are inclusive or open-ended and do not exclude other unlisted elements or method steps.

[0037] As used herein, the terms “one or more” or “at least one” refer to one, two, three, four, five, six, seven, eight, nine or more.

[0038] If the number of components or parts of this invention is not previously specified, it indicates that there is no limitation on the number of times a component or part may appear (or be present). Therefore, it should be interpreted as including one or at least one, and the singular form of a component or part also includes the plural, unless the value clearly indicates a singular number.

[0039] As used herein, the terms “optional” or “optionally” mean that the event or situation described below may or may not occur, including both the occurrence and non-occurrence of the event or situation.

[0040] As used herein, the term "about" means within an acceptable standard error of a value, as would be considered by one of ordinary skill in the art, such as within a 95% confidence interval of the average or within ±10% of a specified value, or a wider range, such as ±0.05, ±0.1, ±0.2, ±0.3, ±1, ±2, or ±3. It should be understood herein that when multiple values ​​or ranges are listed, each listed value or range may be subject to the designation "about," even if only the first value or endpoint is designated as "about."

[0041] In this article, "disodium levonornidazole phosphate" refers to a compound having the following structure.

[0042]

[0043] The term "pharmaceutical composition" refers to an active ingredient that may optionally be combined with one or more pharmaceutically acceptable chemical components (e.g., but not limited to carriers and / or excipients). The active ingredient may be, for example, one or more of the crystals I, II, and III of the present invention.

[0044] The terms “administration” or “giving” refer to methods that enable the delivery of a compound or composition to a desired biological site of action. These methods include, but are not limited to, oral, parenteral (including intravenous, subcutaneous, intraperitoneal, intramuscular, intravascular injection or infusion), local, and rectal administration.

[0045] For pharmaceuticals or pharmacologically active agents, the term "effective amount" refers to a sufficient quantity of a drug or agent that is non-toxic but achieves the desired effect. For the oral dosage forms of this invention, the "effective amount" of one active substance in the composition can be the amount required to achieve the desired effect when used in combination with another active substance in the composition. The determination of the effective amount varies from person to person, depending on the recipient's age and general condition, as well as the specific active substance. A suitable effective amount in any given case can be determined by a person skilled in the art through routine testing.

[0046] The terms "active ingredient," "therapeutic agent," "active substance," or "activator" refer to a chemical entity that can effectively treat or prevent a target disorder, disease, or symptom. In this document, the term may refer to one or more of, for example, Crystal I, Crystal II, and Crystal III of the present invention.

[0047] As used herein, the term "pharmaceutically acceptable carrier" refers to carriers that do not cause significant irritation to the organism and do not impair the biological activity and properties of the active compound. This includes, but is not limited to, any flow aids, sweeteners, diluents, preservatives, dyes / colorants, flavor enhancers, surfactants, wetting agents, dispersants, disintegrants, suspending agents, stabilizers, isotonic agents, solvents, or emulsifiers acceptable for human or animal (e.g., livestock) use. Non-limiting examples of such carriers include calcium carbonate, calcium phosphate, various sugars and starches, cellulose derivatives, gelatin, vegetable oils, and polyethylene glycol. Further information on carriers can be found in Remington: The Science and Practice of Pharmacy, 21st Ed., Lippincott, Williams & Wilkins (2005), the contents of which are incorporated herein by reference.

[0048] As used herein, the term "amorphous" refers to any solid material that is not ordered in three dimensions. In some cases, amorphous solids can be characterized by known techniques, including XRPD crystallography, solid-state nuclear magnetic resonance (ssNMR) spectroscopy, DSC, or some combination of these techniques. As explained below, amorphous solids produce diffuse XRPD spectra, which typically include one or two broad peaks (i.e., peaks with a basis width of about 5°2θ or greater).

[0049] As used herein, the terms “crystal form,” “crystal,” and “crystal shape” have the same meaning and can be used interchangeably in this document. They refer to any solid material exhibiting a three-dimensional arrangement, as opposed to amorphous solid materials, which produce characteristic XRPD spectra with clearly defined peaks.

[0050] As used in the text, the term "solvent compound" refers to a solvent compound formed when a substance dissolves in a solvent, and solvent molecules combine with solute molecules or ions, causing the solute to change its original state.

[0051] As used in this text, the term "hydrate" refers to a salt of the compounds provided herein, which further includes stoichiometric or nonstoichiometric amounts of water bound together by noncovalent intermolecular forces.

[0052] The term "non-covalent form" used in this article refers to weak intermolecular interactions other than covalent bonds, including but not limited to hydrogen bonds, van der Waals forces, salt bonds, hydrophobic interactions, aromatic ring stacking, π-π stacking, halogen bonds, etc.

[0053] As used in this article, the term "X-ray powder diffraction pattern (XRPD pattern)" refers to an experimentally observed diffraction pattern or parameters derived from it. XRPD patterns are typically characterized by peak position (x-axis) and / or peak intensity (y-axis).

[0054] In X-ray powder diffraction (XRPD or XRD) spectroscopy, the diffraction patterns obtained from crystalline compounds are often characteristic of a specific crystal form. The relative intensities of bands (especially at low angles) can vary due to dominant orientation effects resulting from differences in crystallization conditions, grain size, and other measurement conditions. Therefore, the relative intensities of diffraction peaks are not characteristic of the target crystal form; when determining whether a crystal form is identical to a known crystal form, attention should be paid to the relative positions of the peaks rather than their relative intensities. Furthermore, slight errors in peak positions are possible for any given crystal form, which is well known in crystallography. For example, peak positions can shift due to variations in temperature during sample analysis, sample movement, or instrument calibration; the measurement error for 2θ values ​​can sometimes be approximately ±0.2°. Therefore, this error should be taken into account when determining the structure of each crystal form. If the crystal forms of the present invention are described substantially as shown in the specified figures, the term "substantially" is also intended to cover such differences in diffraction peak positions.

[0055] In XRPD spectra, peak positions are typically represented by the 2θ angle or the interplanar distance d, with a simple conversion: d = λ / 2sinθ, where d represents the interplanar distance, λ represents the wavelength of the incident X-rays, and θ is the diffraction angle. For the same crystal form of the same compound, the peak positions in their XRPD spectra are generally similar, although the relative intensity error may be relatively large. It should also be noted that in the identification of mixtures, factors such as decreased content may cause the absence of some diffraction lines. In such cases, it is not necessary to rely on all bands observed in a high-purity sample; even a single band may be characteristic of a given crystal.

[0056] As used herein, the term "2θ" refers to the peak position in degrees, as defined in an experimental setup based on X-ray diffraction experiments, and is typically the horizontal axis unit in a diffraction pattern. If the reflection is diffracted when the incident beam forms an angle θ with a lattice plane, the experimental setup requires recording the reflected beam at a 2θ angle. It should be understood that a specific 2θ value for a particular crystal mentioned herein is intended to represent a 2θ value (in degrees) measured using the X-ray diffraction experimental conditions described herein. X-ray powder diffraction patterns can be obtained by detection under Cu-palladium irradiation conditions using an X-ray powder diffractometer (e.g., an X'Pert3 Powder diffractometer), for example, by detection using an absolute scan at room temperature.

[0057] As used herein, the term "Raman spectroscopy" refers to the signal peaks recorded by a Raman spectrometer. Raman spectroscopy can be performed using a Raman spectrometer (e.g., a Thermo Fisher Nicolet iS50 FT-Raman) at a specific laser wavelength (e.g., 1063 nm) and a specific resolution (e.g., 8 cm⁻¹). -1 The sample is obtained by focusing and scanning under specific sampling gain (e.g., 1.0), moving mirror speed (e.g., 0.3165), and aperture (e.g., 50.00) parameters.

[0058] As used herein, the term "substantially identical" for X-ray diffraction peak positions means taking into account representative peak positions and intensity variations. For example, those skilled in the art will understand that peak positions (2θ) will show some variation, typically up to 0.1–0.2 degrees, and that the instruments used to measure diffraction will also show some variation. Furthermore, those skilled in the art will understand that relative peak intensities will show variations between instruments as well as variations due to degree of crystallinity, preferred orientation, the prepared sample surface, and other factors known to those skilled in the art, and should be considered as qualitative measurements only.

[0059] As used in this article, the term "room temperature" refers to 20℃±5℃.

[0060] When describing methods, components, or steps, the use of letters or numbers for identification is for distinguishing purposes only and does not imply that these methods, components, or steps must be performed in the order or sequence indicated. Those skilled in the art can reasonably adjust these arrangements. For example, "Solvent I," "Solvent II," and "Solvent III" are used for distinguishing purposes only and do not indicate any sequential relationship; the different identifiers may refer to the same or different objects.

[0061] As used in this article, the term "minimum asymmetric unit" refers to the smallest independent part of a unit cell that, after all the space group symmetry operations, can be obtained, which is exactly the smallest part composed of all the atoms, ions, or molecules in the unit cell.

[0062] crystal

[0063] Crystalline form of levonornidazole phosphate disodium pentahydrate I

[0064] This invention relates to crystal I of levonornidazole phosphate disodium pentahydrate, wherein the levonornidazole phosphate disodium pentahydrate has the following structure:

[0065]

[0066] The X-ray powder diffraction pattern of crystal I includes a range of approximately 6.1 ± 0.2. o 15.1±0.2 o 18.3±0.2 o 23.9±0.2 o 24.3±0.2 o and 26.6±0.2 o The diffraction peak at the 2θ angle.

[0067] In a preferred embodiment, the X-ray powder diffraction pattern of crystal I includes approximately 6.1 ± 0.2 o 10.6±0.2 o 11.8±0.2 o 12.4±0.2 o 15.1±0.2 o 15.7±0.2 o 16.0±0.2 o 18.3±0.2 o 20.0±0.2 o 23.9±0.2 o 24.3±0.2 o 26.6±0.2 o and 27.5±0.2 o The diffraction peak at the 2θ angle.

[0068] In a preferred embodiment, the X-ray powder diffraction pattern of crystal I includes approximately 6.1 ± 0.2 o 10.6±0.2 o 11.8±0.2 o 12.4±0.2 o 15.1±0.2 o 15.7±0.2 o 16.0±0.2 o 16.9±0.2 o 17.2±0.2 o 18.3±0.2 o 20.0±0.2 o 23.9±0.2 o 24.3±0.2 o 26.6±0.2 o、 27.5±0.2 o and 30.4±0.2 o The diffraction peak at the 2θ angle.

[0069] In another preferred embodiment, the X-ray powder diffraction pattern of crystal I includes... Figure 1 The peaks at the 2θ angle shown are essentially the same.

[0070] In some embodiments, the X-ray powder diffraction pattern of crystal I includes diffraction peaks at the following 2θ angle:

[0071] <![CDATA[2θ( o )±0.2 o ]]> strength% <![CDATA[2θ( o )±0.2 o ]]> strength% <![CDATA[2θ( o )±0.2 o ]]> strength% <![CDATA[2θ( o )±0.2 o ]]> strength% 6.1 79.46 17.2 7.07 25.4 29.11 32.4 23.42 10.6 35.15 18.3 100.00 26.6 61.63 33.2 19.41 11.8 46.20 20.0 44.37 27.5 44.83 33.7 21.66 12.4 61.78 20.6 22.79 27.8 39.23 34.1 23.05 13.7 20.47 21.3 5.66 28.3 18.18 35.5 12.68 15.1 90.58 22.1 34.24 29.3 3.32 36.3 1.35 15.7 53.94 23.9 72.28 30.4 6.04 37.0 9.32 16.0 68.89 24.3 50.63 30.9 3.26 38.8 3.05 16.9 27.38 25.0 22.31 31.8 28.01 39.4 4.49

[0072] In a more preferred embodiment, the X-ray powder diffraction pattern of crystal I is as follows: Figure 1 As shown.

[0073] In some implementations, the moisture content of crystal form I is determined to be 21.1% using the Karl Fischer method.

[0074] In some implementation schemes, the crystallographic structure data and structural refinement parameters of crystal I are as follows:

[0075]

[0076]

[0077] Crystals of levonornidazole phosphate disodium hexahydrate II

[0078] This invention provides crystal II of levonornidazole phosphate disodium hexahydrate, wherein the levonornidazole phosphate disodium hexahydrate has the following structure:

[0079] The X-ray powder diffraction pattern of crystal II includes approximately 6.0 ± 0.2 o 15.0±0.2 o 15.8±0.2 o 16.9±0.2 o 24.0±0.2 o and 27.0±0.2 o The diffraction peak at the 2θ angle.

[0080] In a preferred embodiment, the X-ray powder diffraction pattern of crystal II includes approximately 6.0 ± 0.2 nm. o 10.6±0.2 o 12.4±0.2 o 15.0±0.2 o 15.8±0.2 o 16.1±0.2 o 16.9±0.2 o 18.2±0.2 o 20.5±0.2 o 24.0±0.2 o 25.5±0.2 o and 27.0±0.2 o The diffraction peak at the 2θ angle.

[0081] In a preferred embodiment, the X-ray powder diffraction pattern of crystal II includes approximately 6.0 ± 0.2 nm. o 10.6±0.2 o 12.4±0.2 o 15.0±0.2 o 15.8±0.2 o 16.1±0.2 o 16.9±0.2 o 18.2±0.2 o 20.5±0.2 o 22.4±0.2 o 24.0±0.2 o 24.4±0.2 o 24.6±0.2 o 25.5±0.2 o 25.8±0.2 o 27.0±0.2 o and 35.9±0.2 o The diffraction peak at the 2θ angle.

[0082] In a preferred embodiment, the X-ray powder diffraction pattern of crystal II includes approximately 6.0 ± 0.2 nm. o10.6±0.2 o 12.4±0.2 o 15.0±0.2 o 15.8±0.2 o 16.1±0.2 o 16.9±0.2 o 18.2±0.2 o 20.5±0.2 o 22.4±0.2 o 24.0±0.2 o 24.4±0.2 o 24.6±0.2 o 25.5±0.2 o 25.8±0.2 o 27.0±0.2 o 31.1±0.2 o 31.5±0.2 o 35.9±0.2 o The diffraction peak at the 2θ angle.

[0083] In another preferred embodiment, the X-ray powder diffraction pattern of crystal II includes... Figure 2 The peaks at the 2θ angle are essentially the same as those shown.

[0084] In some embodiments, the X-ray powder diffraction pattern of crystal II includes diffraction peaks at the following 2θ angle:

[0085]

[0086]

[0087] In a more preferred embodiment, the X-ray powder diffraction pattern of crystal II is as follows: Figure 2 As shown.

[0088] In some implementations, the moisture content of crystal form II is determined to be 24.0% using the Karl Fischer method.

[0089] In some implementation schemes, crystal II obtains crystallographic structure data and structural refinement parameters through microcrystalline electron diffraction as follows:

[0090]

[0091]

[0092] Levonidazole phosphate disodium hexahydrate crystals III

[0093] This invention provides crystal III of levonornidazole phosphate disodium hexahydrate, wherein the levonornidazole phosphate disodium hexahydrate has the following structure:

[0094]

[0095] The X-ray powder diffraction pattern of crystal III includes approximately 6.0 ± 0.2 o 11.7±0.2 o 12.1±0.2 o 15.7±0.2 o 19.6±0.2 o and 31.9±0.2 o The diffraction peak at the 2θ angle.

[0096] In a preferred embodiment, the X-ray powder diffraction pattern of crystal III includes approximately 6.0 ± 0.2 nm. o 11.7±0.2 o 12.1±0.2 o 15.7±0.2 o 16.8±0.2 o 17.0±0.2 o 19.6±0.2 o 20.2±0.2 o 26.4±0.2 o 26.7±0.2 o 28.7±0.2 o and 31.9±0.2 o The diffraction peak at the 2θ angle.

[0097] In a preferred embodiment, the X-ray powder diffraction pattern of crystal III includes approximately 6.0 ± 0.2 nm. o 11.7±0.2 o 12.1±0.2 o 15.7±0.2 o 16.8±0.2 o 17.0±0.2 o 17.4±0.2 o 19.6±0.2 o 20.2±0.2 o 23.6±0.2 o 24.0±0.2 o 26.4±0.2 o 26.7±0.2 o 28.7±0.2 o and 31.9±0.2 o The diffraction peak at the 2θ angle.

[0098] In a preferred embodiment, the X-ray powder diffraction pattern of crystal III includes approximately 6.0 ± 0.2 nm. o 11.7±0.2 o 12.1±0.2 o 12.4±0.2 o 15.7±0.2 o 16.8±0.2 o 17.0±0.2 o 17.4±0.2 o 19.6±0.2 o 20.2±0.2 o 23.6±0.2 o 24.0±0.2 o 26.4±0.2 o 26.7±0.2 o 28.7±0.2 o 31.9±0.2 o 34.4±0.2 o and 34.8±0.2 o The diffraction peak at the 2θ angle.

[0099] In another preferred embodiment, the X-ray powder diffraction pattern of crystal III includes... Figure 3 The peaks at the 2θ angle are essentially the same as those shown.

[0100] In some embodiments, the X-ray powder diffraction pattern of crystal III includes diffraction peaks at the following 2θ angle:

[0101]

[0102]

[0103] In a more preferred embodiment, the X-ray powder diffraction pattern of crystal III is as follows: Figure 3 As shown.

[0104] In some implementations, the water content of crystal form III is determined to be 24.2% using the Karl Fischer method.

[0105] In some implementations, crystal III obtains crystallographic structure data and structural refinement parameters via microcrystalline electron diffraction (MicroED) as follows:

[0106]

[0107] Crystal preparation methods

[0108] Another object of the present invention is to provide a method for preparing the crystal of the present invention.

[0109] Preparation method of crystal I of levonornidazole phosphate disodium pentahydrate

[0110] This invention provides a method for preparing crystal I of levonornidazole phosphate disodium pentahydrate, comprising the following steps:

[0111] Disodium levonornidazole phosphate and solvent I are mixed, heated to 40-50℃ to dissolve, filtered, and the filtrate is cooled to crystallize, thus obtaining crystal I;

[0112] The solvent I is selected from water, ethanol, isopropanol, acetone or acetonitrile.

[0113] In some embodiments, solvent I is selected from ethanol, such as 90-95% ethanol, 90% ethanol, 93% ethanol, and 95% ethanol.

[0114] In some embodiments, the cooling crystallization is achieved by cooling to -10°C to room temperature, for example by cooling to room temperature and stirring to induce crystallization, or by cooling to -10°C and allowing the crystals to stand to induce crystallization.

[0115] In some embodiments, the mass-to-volume ratio of disodium levonornidazole phosphate to solvent I, in g / mL, is 1:5-50, for example 1:5-30, 1:5-25, 1:10-25, 1:10-20, 1:10, 1:12, 1:15, 1:18, 1:20, 1:22, 1:25, 1:30, 1:35, 1:40, 1:45, 1:50.

[0116] Preparation method of crystals of levonornidazole phosphate disodium hexahydrate II

[0117] This invention provides a method for preparing crystal II of levonornidazole phosphate disodium hexahydrate, comprising the following steps:

[0118] Disodium levonornidazole phosphate and solvent II are mixed, heated to 40-50℃ to dissolve, filtered, and the filtrate is cooled to crystallize, thus obtaining crystal II;

[0119] The solvent II is selected from a mixture of at least two of water, ethanol, propanol, or acetone.

[0120] In some embodiments, solvent II is selected from a mixture of water / acetone / ethanol.

[0121] In some embodiments, solvent II is selected from a mixed solvent of water / acetone / ethanol with a volume ratio of 1-5:5-10:5-10, for example, a mixed solvent of water / acetone / ethanol with a volume ratio of 2:9:9.

[0122] In some embodiments, the mass-to-volume ratio of the disodium levonornidazole phosphate to solvent II, in g / mL, is 1:5-50, for example 1:5-30, 1:5-25, 1:10-25, 1:10-20, 1:5, 1:8, 1:10, 1:12, 1:15, 1:18, 1:20, 1:22, 1:25, 1:28, or 1:30.

[0123] In some embodiments, the cooling crystallization is achieved by cooling to -10°C to room temperature, for example by cooling to room temperature and stirring to induce crystallization, or by cooling to -10°C and allowing the crystals to stand to induce crystallization.

[0124] Preparation method of crystals III of levonornidazole phosphate disodium hexahydrate

[0125] This invention provides a method for preparing crystal III of levonornidazole phosphate disodium hexahydrate, comprising the following steps:

[0126] Disodium levonornidazole phosphate and solvent III are mixed, heated to 40-50℃ to dissolve, filtered, the filtrate is cooled to 30-35℃, then cooled to crystallize, filtered, and the filter cake is washed with acetone to obtain crystal III;

[0127] The solvent III is selected from a mixture of at least two of water, ethanol, propanol, or acetone.

[0128] In some embodiments, solvent III is selected from a mixture of water / acetone / ethanol.

[0129] In some embodiments, solvent III is selected from a mixed solvent of water / acetone / ethanol with a volume ratio of 1-5:5-10:5-10, for example, a mixed solvent of water / acetone / ethanol with a volume ratio of 2:9:9.

[0130] In some embodiments, the mass-to-volume ratio of the disodium levonornidazole phosphate to solvent III, in g / mL, is 1:5-50, for example 1:5-30, 1:5-25, 1:10-25, 1:10-20, 1:5, 1:8, 1:10, 1:12, 1:15, 1:18, 1:20, 1:22, 1:25, 1:28, or 1:30.

[0131] In some embodiments, the cooling crystallization is achieved by cooling to -10°C to room temperature, for example by cooling to room temperature and stirring to induce crystallization, or by cooling to -10°C and allowing the crystals to stand to induce crystallization.

[0132] In some embodiments, the preparation method includes the following steps:

[0133] Disodium levonidazole phosphate and solvent III are mixed, heated to 40-50℃ to dissolve, filtered, and seed crystals are added after the filtrate is cooled to 30-35℃. Crystals are then precipitated by cooling, filtered, and the filter cake is washed with acetone to obtain crystal III.

[0134] Pharmaceutical Compositions and Uses

[0135] Another object of the present invention is to provide a pharmaceutical composition comprising i) any one or more of the crystals of the present invention: crystal I of levonornidazole phosphate disodium pentahydrate, crystal II of levonornidazole phosphate disodium hexahydrate, crystal III of levonornidazole phosphate disodium hexahydrate, and ii) one or more pharmaceutically acceptable carriers.

[0136] Another object of the present invention is to provide the use of the crystals of the present invention (e.g., crystal I of levonornidazole phosphate disodium pentahydrate, crystal II of levonornidazole phosphate disodium hexahydrate, crystal III of levonornidazole phosphate disodium hexahydrate, or any combination thereof) or pharmaceutical compositions thereof in the preparation of drugs for anti-anaerobic bacterial infections and drugs for antiprotozoal infections.

[0137] Another object of the present invention is to provide the crystals (e.g., crystal I of levonornidazole phosphate disodium pentahydrate, crystal II of levonornidazole phosphate disodium hexahydrate, crystal III of levonornidazole phosphate disodium hexahydrate, or any combination thereof) or pharmaceutical compositions of the present invention for use against anaerobic bacterial infections and antiprotozoal infections.

[0138] Another object of the present invention is to provide a method for treating anaerobic bacterial infections and antiprotozoal infections, comprising administering to an individual in need a therapeutically effective amount of the crystals of the present invention (e.g., crystal I of levonornidazole phosphate disodium pentahydrate, crystal II of levonornidazole phosphate disodium hexahydrate, crystal III of levonornidazole phosphate disodium hexahydrate, or any combination thereof).

[0139] The crystals of the present invention can be administered, either in their pure form or as a suitable pharmaceutical composition, via any acceptable mode of administration that provides a medicine for similar use. The pharmaceutical compositions of the present invention can be prepared by combining the crystal form of the present invention with a suitable pharmaceutically acceptable carrier.

[0140] The pharmaceutical compositions of the present invention can be manufactured using methods well known in the art, such as conventional mixing methods, etc.

[0141] Typical routes of administration for the crystals or pharmaceutical compositions of the present invention include, but are not limited to, oral, rectal, transmucosal, enteral, or topical, transdermal, inhalation, parenteral, sublingual, intravaginal, intranasal, intraocular, intraperitoneal, intramuscular, subcutaneous, and intravenous administration.

[0142] In a preferred embodiment, the pharmaceutical composition is in oral form. For oral administration, the pharmaceutical composition can be formulated by mixing the active compound with a pharmaceutically acceptable carrier, excipient, and / or medium well known in the art. These carriers, excipients, and media enable the compounds of the present invention to be formulated into tablets, pills, lozenges, sugar-coated tablets, capsules, liquids, gels, pastes, suspensions, etc., for oral administration to patients.

[0143] Solid oral compositions can be prepared using conventional mixing, filling, or tableting methods. For example, they can be obtained by mixing the active compound with a solid excipient, optionally milling the resulting mixture, adding other suitable excipients if necessary, and then processing the mixture into granules to obtain the core of a tablet or sugar-coated formulation.

[0144] Example

[0145] The present invention will be further illustrated by the following embodiments. The embodiments of the present invention are only used to illustrate the technical solutions of the present invention and are not intended to limit the scope of the present invention. Those skilled in the art can make some non-essential improvements and adjustments, which still fall within the protection scope of the present invention.

[0146] Information on the testing instruments and methods used in the experiment:

[0147] The testing conditions for X-ray powder diffraction (XRPD) are as follows:

[0148] An X'Pert3 powder diffractometer was used, which employs Cu-palladium irradiation and Absolutescan detection at room temperature. The detection range was 3.5° to 40°, with a step size of 0.013, a dwell time of 50 s, and one scan.

[0149] The Raman spectroscopy testing conditions were as follows: A Thermo Fisher Nicoleti S50 FT-Raman instrument was used, which directly focused and scanned the sample using a laser wavelength of 1063 nm, with a resolution of 8 cm⁻¹. -1 The number of scans was 128, the sampling gain was 1.0, the moving mirror speed was 0.3165, and the aperture was 50.00.

[0150] The liquid chromatography instrument used for detection was an Agilent 1260 HPLC.

[0151] The HPLC experimental conditions are as follows:

[0152] Column: Capcell Pak MGⅡC18, 4.6mm × 250mm, 5μm

[0153] Flow rate: 1.0 ml / min, column temperature: 25±5℃, detection wavelength: 321 nm;

[0154] Mobile phase A: 6.8 g / L potassium dihydrogen phosphate solution (pH 6.5);

[0155] Mobile phase B: Methanol

[0156] Gradient elution: 0-50 min, volume percentage of mobile phase A% : mobile phase B% = (90-40) : (10-60); 50-70 min, volume percentage of mobile phase A% : mobile phase B% = (40-90) : (60-10).

[0157] All solvents used in this invention are commercially available and can be used without further purification.

[0158] Example 1. Preparation of crystal I of levonornidazole phosphate disodium pentahydrate

[0159] Weigh 20g of disodium levonornidazole phosphate, add 500ml of 95% ethanol, stir and heat to 45℃ to dissolve. After the solution is clear, filter, slowly cool to -10℃, stir to precipitate crystals for 4 hours, filter again, collect the filter cake, dry at room temperature for 2 hours, and collect the solid. Perform XRPD analysis on the solid. The obtained XRPD spectrum is shown below. Figure 1 As shown, this is crystal I of levonornidazole phosphate disodium pentahydrate.

[0160] Example 2. Preparation of single crystal I of levonornidazole phosphate disodium pentahydrate

[0161] Weigh 1g of disodium levonornidazole phosphate, add 10ml of 93% ethanol, stir and heat to 50℃ to dissolve, filter through a filter membrane, and allow the filtrate to crystallize at room temperature for 1 day to obtain crystal I single crystal sample. Single crystal diffraction tests were performed on the sample, and its single crystal structure schematic diagram is shown below. Figure 4 The sample was subjected to XRPD detection, and the obtained XRPD spectrum was basically consistent with that of Example 1, which is crystal I of levonornidazole phosphate disodium pentahydrate.

[0162] Example 3. Preparation of crystal II of levonornidazole phosphate disodium hexahydrate

[0163] Weigh 10g of disodium levonornidazole phosphate and add it to 200ml of a water / acetone / ethanol (2:9:9, v / v / v) mixed solvent. Stir and heat to 45℃ to dissolve. After the solution is clear, filter and cool to -10℃ to crystallize for 4 hours. Filter again, collect the filter cake, and dry it at 30℃ for 2 hours. Collect the solid. Perform XRPD analysis on the solid. The obtained XRPD spectrum is shown below. Figure 2 As shown, this is crystal II of levonornidazole phosphate disodium hexahydrate.

[0164] Example 4. Preparation of crystals III of levonornidazole phosphate disodium hexahydrate

[0165] Weigh 10g of disodium levonornidazole phosphate and add it to 200ml of a water / acetone / ethanol (2:9:9, v / v / v) mixture. Stir and heat to 45℃ to dissolve. After the solution is clear, filter. Cool to 33℃, add seed crystals, and continue cooling to -10℃ to crystallize for 4 hours. Filter and wash the filter cake with acetone. Collect the filter cake and dry it at 30℃ for 2 hours. Perform XRPD analysis on the solid. The obtained XRPD spectrum is shown below. Figure 3 As shown, this is crystal III of levonornidazole phosphate disodium hexahydrate.

[0166] Experimental Example 1. Crystal Form Stability Test

[0167] The crystal I obtained in Example 1, the crystal II obtained in Example 3, and the crystal III obtained in Example 4, as well as the crystal obtained in Example 1 according to CN201910115156.7, were subjected to stability tests under the conditions shown in the table below. The results show that crystal I, crystal II, and crystal III of the present invention are relatively stable at both room temperature and low temperature. The results are shown in Tables 1-1 and 1-2 below:

[0168] Table 1-1: Stability test results of crystal form I

[0169]

[0170]

[0171] Table 1-2: Stability test results of crystal form II and crystal form III

[0172]

[0173] Experimental Example 3. Powder Properties Test

[0174] Microscopic observation of the crystal morphology of crystals I, II, and III is shown in the following microscopic images. Figure 7-9 As shown, crystal I is rod-shaped, crystal II is plate-shaped, and crystal III is a larger cuboid crystal. Based on crystal morphology, crystal III exhibits better physical properties.

[0175] The flowability of crystals I, II and III of compound (I) was evaluated according to the "Determination of bulk density and tap density" in Pharmacopoeia 0093 of 2020.

[0176] Analysis shows that the crystals I, II, and III of the compound of formula (I) of this invention have excellent flowability, which is effective in improving industrial operability and productivity. The product quality is uniform, and there is less adhesion to the grinding mill during crystallization and crushing. The charge is low, and it has excellent operation performance.

[0177] The above detailed embodiments provide a further description of the present invention. However, it should not be construed as limiting the scope of the present invention to the listed embodiments; all technical solutions implemented based on the content of the present invention fall within the scope of the present invention.

Claims

1. Crystalline form III of levonornidazole phosphate disodium hexahydrate, characterized in that, The levonornidazole phosphate disodium hexahydrate has the following structure: The X-ray powder diffraction pattern of crystal III includes approximately 6.0 ± 0.2 o 11.7±0.2 o 12.1±0.2 o 15.7±0.2 o 19.6±0.2 o and 31.9±0.2 o The diffraction peak at the 2θ angle; Preferably, the X-ray powder diffraction pattern of crystal III includes a range of approximately 6.0 ± 0.

2. o 11.7±0.2 o 12.1±0.2 o 15.7±0.2 o 16.8±0.2 o 17.0±0.2 o 19.6±0.2 o 20.2±0.2 o 26.4±0.2 o 26.7±0.2 o 28.7±0.2 o and 31.9±0.2 o The diffraction peak at the 2θ angle; Preferably, the X-ray powder diffraction pattern of crystal III includes a range of approximately 6.0 ± 0.

2. o 11.7±0.2 o 12.1±0.2 o 15.7±0.2 o 16.8±0.2 o 17.0±0.2 o 17.4±0.2 o 19.6±0.2 o 20.2±0.2 o 23.6±0.2 o 24.0±0.2 o 26.4±0.2 o 26.7±0.2 o 28.7±0.2 o and 31.9±0.2 o The diffraction peak at the 2θ angle; Preferably, the X-ray powder diffraction pattern of crystal III includes a range of approximately 6.0 ± 0.

2. o 11.7±0.2 o 12.1±0.2 o 12.4±0.2 o 15.7±0.2 o 16.8±0.2 o 17.0±0.2 o 17.4±0.2 o 19.6±0.2 o 20.2±0.2 o 23.6±0.2 o 24.0±0.2 o 26.4±0.2 o 26.7±0.2 o 28.7±0.2 o 31.9±0.2 o 34.4±0.2 o and 34.8±0.2 o The diffraction peak at the 2θ angle.

2. The crystal III of claim 1, characterized in that, The X-ray powder diffraction pattern of the crystal III includes a peak at the 2θ angle that is essentially the same as that shown in Figure 3; Preferably, the X-ray powder diffraction pattern of crystal III includes the following diffraction peaks at the 2θ angle: Preferably, the X-ray powder diffraction pattern of crystal III is shown in Figure 3; Preferably, the crystallographic structure data and structural refinement parameters of crystal form III obtained by microcrystalline electron diffraction are as follows: Crystal system: monoclinic; Space group: P21 (No. 4); Cell parameters: α=90°, β=92(2)°, γ=90°.

3. Crystalline form I of levonornidazole phosphate disodium pentahydrate, characterized in that, The levonornidazole phosphate disodium pentahydrate has the following structure: The X-ray powder diffraction pattern of crystal I includes a range of approximately 6.1 ± 0.

2. o 15.1±0.2 o 18.3±0.2 o 23.9±0.2 o 24.3±0.2 o and 26.6±0.2 o The diffraction peak at the 2θ angle; Preferably, the X-ray powder diffraction pattern of crystal I includes a range of approximately 6.1 ± 0.2 nm. o 10.6±0.2 o 11.8±0.2 o 12.4±0.2 o 15.1±0.2 o 15.7±0.2 o 16.0±0.2 o 18.3±0.2 o 20.0±0.2 o 23.9±0.2 o 24.3±0.2 o 26.6±0.2 o and 27.5±0.2 o The diffraction peak at the 2θ angle; Preferably, the X-ray powder diffraction pattern of crystal I includes a range of approximately 6.1 ± 0.2 nm. o 10.6±0.2 o 11.8±0.2 o 12.4±0.2 o 15.1±0.2 o 15.7±0.2 o 16.0±0.2 o 16.9±0.2 o 17.2±0.2 o 18.3±0.2 o 20.0±0.2 o 23.9±0.2 o 24.3±0.2 o 26.6±0.2 o、 27.5±0.2 o and 30.4±0.2 o The diffraction peak at the 2θ angle.

4. The crystal I according to claim 3, characterized in that, The X-ray powder diffraction pattern of crystal I includes a peak at the 2θ angle that is essentially the same as that shown in Figure 1. Preferably, the X-ray powder diffraction pattern of crystal I includes the following diffraction peaks at the 2θ angle: Preferably, the X-ray powder diffraction pattern of crystal I is shown in Figure 1.

5. Crystal form I according to claim 3 or 4, wherein the crystallographic structure data and structural refinement parameters of crystal I are as follows: the crystal system is orthorhombic, the space group is P212121, and the unit cell size is as follows: α = 90° β = 90° γ = 90°; Z value is 4, density is 1.555 g / cm³ 3 The number of electrons F in a single cell is 896.

0.

6. Crystalline form II of levonornidazole phosphate disodium hexahydrate, characterized in that, The levonornidazole phosphate disodium hexahydrate has the following structure: The X-ray powder diffraction pattern of crystal II includes approximately 6.0 ± 0.2 o 15.0±0.2 o 15.8±0.2 o 16.9±0.2 o 24.0±0.2 o and 27.0±0.2 o The diffraction peak at the 2θ angle; Preferably, the X-ray powder diffraction pattern of crystal II includes a range of approximately 6.0 ± 0.

2. o 10.6±0.2 o 12.4±0.2 o 15.0±0.2 o 15.8±0.2 o 16.1±0.2 o 16.9±0.2 o 18.2±0.2 o 20.5±0.2 o 24.0±0.2 o 25.5±0.2 o and 27.0±0.2 o The diffraction peak at the 2θ angle; Preferably, the X-ray powder diffraction pattern of crystal II includes a range of approximately 6.0 ± 0.

2. o 10.6±0.2 o 12.4±0.2 o 15.0±0.2 o 15.8±0.2 o 16.1±0.2 o 16.9±0.2 o 18.2±0.2 o 20.5±0.2 o 22.4±0.2 o 24.0±0.2 o 24.4±0.2 o 24.6±0.2 o 25.5±0.2 o 25.8±0.2 o 27.0±0.2 o and 35.9±0.2 o The diffraction peak at the 2θ angle; Preferably, the X-ray powder diffraction pattern of crystal II includes a range of approximately 6.0 ± 0.

2. o 10.6±0.2 o 12.4±0.2 o 15.0±0.2 o 15.8±0.2 o 16.1±0.2 o 16.9±0.2 o 18.2±0.2 o 20.5±0.2 o 22.4±0.2 o 24.0±0.2 o 24.4±0.2 o 24.6±0.2 o 25.5±0.2 o 25.8±0.2 o 27.0±0.2 o 31.1±0.2 o 31.5±0.2 o 35.9±0.2 o The diffraction peak at the 2θ angle.

7. The crystal II according to claim 6, characterized in that... The X-ray powder diffraction pattern of crystal II includes a peak at the 2θ angle that is essentially the same as that shown in Figure 2; Preferably, the X-ray powder diffraction pattern of crystal II includes the following diffraction peaks at the 2θ angle: Preferably, the X-ray powder diffraction pattern of crystal II is shown in Figure 2; Preferably, the crystallographic structure data and structural refinement parameters for crystal form II obtained by microcrystalline electron diffraction are as follows: Crystal system: orthorhombic; Space group: P212121 (No. 19); Cell parameters: α=90°, β=90°, γ=90°.

8. The method for preparing crystal I of levonornidazole phosphate disodium pentahydrate according to any one of claims 3-5, characterized in that, Includes the following steps: Disodium levonornidazole phosphate and solvent I are mixed, heated to 40-50℃ to dissolve, filtered, and the filtrate is cooled to crystallize, thus obtaining crystal I; The solvent I is selected from water, ethanol, acetone or acetonitrile.

9. The method for preparing crystal I of levonornidazole phosphate disodium pentahydrate according to claim 8, characterized in that... One or more of the following: (1) The solvent I is selected from ethanol, such as 90-95% ethanol, 90% ethanol, 93% ethanol, and 95% ethanol; (2) The cooling crystallization is cooling to -10℃ to room temperature, for example, cooling to room temperature and stirring to crystallize, or cooling to -10℃ and letting it stand to crystallize; (3) The mass-volume ratio of disodium levonidazole phosphate to solvent I, in g / mL, is 1:5-50, for example 1:5-30, 1:5-25, 1:10-25, 1:10-20.

10. The method for preparing crystal II of levonornidazole phosphate disodium hexahydrate according to claim 6 or 7, characterized in that, Includes the following steps: Disodium levonornidazole phosphate and solvent II are mixed, heated to 40-50℃ to dissolve, filtered, and the filtrate is cooled to crystallize, thus obtaining crystal II; The solvent II is selected from a mixture of at least two of water, ethanol, propanol, or acetone.

11. The method for preparing crystal II of levonornidazole phosphate disodium hexahydrate according to claim 10, characterized in that... One or more of the following: (1) Solvent II is selected from a mixed solvent of water / acetone / ethanol; (2) The solvent II is selected from a mixed solvent of water / acetone / ethanol with a volume ratio of 1-5:5-10:5-10, for example, a mixed solvent of water / acetone / ethanol with a volume ratio of 2:9:

9. (3) The mass-volume ratio of the disodium levonidazole phosphate to solvent II, in g / mL, is 1:5-50, for example 1:5-30, 1:5-25, 1:10-25, 1:10-20; (4) The cooling crystallization is cooling to -10℃ to room temperature, for example cooling to room temperature and stirring to crystallize, or cooling to -10℃ and letting it stand to crystallize.

12. A method for preparing crystal III of levonornidazole phosphate disodium hexahydrate according to claim 1 or 2, comprising the following steps: Disodium levonornidazole phosphate and solvent III are mixed, heated to 40-50℃ to dissolve, filtered, the filtrate is cooled to 30-35℃, then cooled to crystallize, filtered, and the filter cake is washed with acetone to obtain crystal III. The solvent III is selected from a mixture of at least two of water, ethanol, propanol, or acetone.

13. The method for preparing crystal III of levonornidazole phosphate disodium hexahydrate according to claim 12, characterized in that... One or more of the following: (1) Solvent III is selected from a mixed solvent of water / acetone / ethanol; (2) The solvent III is selected from a mixed solvent of water / acetone / ethanol with a volume ratio of 1-5:5-10:5-10, for example, a mixed solvent of water / acetone / ethanol with a volume ratio of 2:9:

9. (3) The mass-volume ratio of the disodium levonidazole phosphate to solvent III, in g / mL, is 1:5-50, for example 1:5-30, 1:5-25, 1:10-25, 1:10-20; (4) The cooling crystallization is cooling to -10℃ to room temperature, for example, cooling to room temperature and stirring to crystallize, or cooling to -10℃ and letting it stand to crystallize; (5) The preparation method includes the following steps: mixing disodium levofloxacin phosphate and solvent III, heating to 40-50℃ to dissolve, filtering, cooling the filtrate to 30-35℃ and adding seed crystals, then cooling to precipitate crystals, filtering, and washing the filter cake with acetone to obtain crystal III.

14. A pharmaceutical composition comprising crystal I of levonornidazole phosphate disodium pentahydrate as claimed in any one of claims 3-5, crystal II of levonornidazole phosphate disodium hexahydrate as claimed in claim 6 or 7, or crystal III of levonornidazole phosphate disodium hexahydrate as claimed in claim 1 or 2, and one or more pharmaceutically acceptable carriers.

15. Use of crystal I of levonornidazole phosphate disodium pentahydrate according to any one of claims 3-5, crystal II of levonornidazole phosphate disodium hexahydrate according to claim 6 or 7, crystal III of levonornidazole phosphate disodium hexahydrate according to claim 1 or 2, or the pharmaceutical composition according to claim 14, in the preparation of drugs for anti-anaerobic bacterial infection and anti-protozoal infection.

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