Ensifentrine-saccharin solid form, preparation method therefor, and use thereof

Abstract

The present invention relates to an Ensifentrine-saccharin solid form having a structure as shown in formula I. The solid form can effectively improve the solubility of Ensifentrine to enhance bioavailability and achieve good physicochemical stability, and is widely applicable to the development of improved formulations.

Description

Encerfentin - Saccharin Solid Form, Preparation Method and Uses

[0001] This application requires the applicant to have:

[0002] Priority rights to the earlier application filed with the China National Intellectual Property Administration on December 20, 2024, with patent application number 202411889726.5 and entitled "Ensefentin - Saccharin Solid Form and Preparation Method and Use thereof";

[0003] The full text of the prior application is incorporated herein by reference. Technical Field

[0004] This invention belongs to the field of pharmaceutical compounds, specifically relating to the solid form of ensefenine-saccharin, its preparation method, and its uses. Background Technology

[0005] Ensifentrine (trade name: Ohtuvayre) is a small molecule drug approved by the FDA on June 26, 2024, for the treatment of chronic obstructive pulmonary disease (COPD) in adults. This drug has a dual inhibitory effect on PDE3 and PDE4; inhibition of PDE3 modulates airway smooth muscle, thereby dilating the airways; while inhibition of PDE4 participates in the inflammatory cell activation and migration of bronchial epithelial cells, and activates transmembrane transduction regulators in cystic fibrosis to reduce mucus viscosity and improve mucociliary clearance. This dual inhibition shows a synergistic effect in airway smooth muscle contraction and inflammation control, thus providing a new non-steroidal anti-inflammatory treatment option for COPD patients. Ohtuvayre is currently administered via inhalation after being nebulized as a suspension.

[0006] Existing literature (Ananya Kar, et al., CrystEngComm, 2024, 26, 3783) reports that ensifentrine exists in three crystal forms: crystal form I, crystal form II, and crystal form III. Among them, crystal form I is the crystal form used in ensifentrine nebulized inhalation formulations; however, the water solubility of this crystal form is only 29 μg / mL, which greatly limits the bioavailability of ensifentrine. Therefore, it is necessary to develop a solid form of ensifentrine that is more suitable for pharmaceutical use. Summary of the Invention

[0007] To address the problems existing in the prior art, on the one hand, the present invention provides an encefentin-saccharin solid form, which has the structure shown in Formula I below:

[0008] In some embodiments, the solid form is encefenstein-saccharin solid form A, and the X-ray powder diffraction pattern of encefenstein-saccharin solid form A, expressed as a diffraction angle of 2θ±0.2°, shows diffraction peaks at 10.29°, 11.12°, and 19.81°; furthermore, the X-ray powder diffraction pattern of encefenstein-saccharin solid form A, expressed as a diffraction angle of 2θ±0.2°, also shows diffraction peaks at one or more of the following locations: 8.34°, 10.89°, 21.92°, 22.45°, and 23.61°. Furthermore, the X-ray powder diffraction pattern of the encefenstein-saccharin solid form A, expressed as a diffraction angle of 2θ±0.2°, also has diffraction peaks at one or more of the following locations: 11.69°, 13.30°, 15.03°, 15.50°, 19.10°, 25.59°, and 28.38°. Furthermore, the encefenstein-saccharin solid form A has an XRPD pattern basically as shown in Figure 1. Furthermore, the encefenstein-saccharin solid form A has one or more of the diffraction peaks listed in Table A-1.

[0009] In some embodiments, the encefentin-saccharin solid form A has one or more of the following characteristics:

[0010] (1) The TGA curve of ensifine-saccharin solid form A showed a weight loss of 0-5% at 180.39±5℃, for example, a weight loss of 3.34%;

[0011] (2) The DSC curve of Ensefentin-saccharin solid form A shows that melting occurs at the onset point of 209.29±5℃, and the peak melting temperature is 212.77±5℃.

[0012] In some embodiments, the TGA / DSC plot of the encefentin-saccharin solid form A is basically as shown in Figure 3; the encefentin-saccharin solid form A... 1 The basic H NMR spectrum is shown in Figure 4.

[0013] In some embodiments, the encefenstein-saccharin solid form A has the following crystallographic parameters: chiral space group P21 / n; and unit cell parameters: α=90°, β=91.662(3)°, γ=90°, The number of molecules (Z) in a unit lattice is 4, and the density (D) is 1.374 g / cm³. 3 Furthermore, the crystallographic parameters of the encefentin-saccharin solid morphology A are shown in Table A-2.

[0014] According to an embodiment of the present invention, the encefentin-saccharin solid form A is a solvent-free substance, wherein one molecule of encefentin and one molecule of saccharin are combined to form a solid form.

[0015] In some embodiments, the solid form is encefenstein-saccharin solid form B, and the X-ray powder diffraction pattern of encefenstein-saccharin solid form B, expressed as a diffraction angle of 2θ±0.2°, shows diffraction peaks at 10.32°, 11.12°, and 19.58°; furthermore, the X-ray powder diffraction pattern of encefenstein-saccharin solid form B, expressed as a diffraction angle of 2θ±0.2°, also shows diffraction peaks at one or more of 8.19°, 15.13°, and 23.23°; even further, the encefenstein-saccharin solid form B... The X-ray powder diffraction pattern of the saccharin-saccharin solid form B, expressed as a diffraction angle of 2θ±0.2°, also shows diffraction peaks at one or more of the following locations: 11.69°, 17.00°, 18.62°, 20.97°, 21.69°, 22.34°, 25.09°, and 25.50°. Furthermore, the saccharin-saccharin solid form B has an XRPD pattern similar to that shown in Figure 7. Even further, the saccharin-saccharin solid form B has one or more of the diffraction peaks listed in Table A-3.

[0016] In some embodiments, the solid form is encefenstein-saccharin solid form C, and the X-ray powder diffraction pattern of encefenstein-saccharin solid form C, expressed as a diffraction angle of 2θ±0.2°, has diffraction peaks at 15.14°, 19.36°, and 22.96°; furthermore, the X-ray powder diffraction pattern of encefenstein-saccharin solid form C, expressed as a diffraction angle of 2θ±0.2°, also has diffraction peaks at one or more of 10.29°, 11.08°, and 22.43°; even further, the X-ray powder diffraction pattern of encefenstein-saccharin solid form C, expressed as a diffraction angle of 2θ±0.2°, also has peaks at 8°. The encefenstein-saccharin solid form C exhibits diffraction peaks at one or more of the following angles: 0.7°, 13.14°, 18.67°, and 22.18°. Furthermore, the X-ray powder diffraction pattern of the encefenstein-saccharin solid form C, expressed as a diffraction angle of 2θ ± 0.2°, also exhibits diffraction peaks at one or more of the following angles: 11.65°, 19.70°, 20.95°, 21.17°, 21.50°, 25.07°, and 25.41°. Further, the encefenstein-saccharin solid form C has an XRPD pattern essentially as shown in Figure 11. Even further, the encefenstein-saccharin solid form C has one or more diffraction peaks listed in Table A-4.

[0017] In some embodiments, the solid form is encefenstein-saccharin solid form D, and the X-ray powder diffraction pattern of encefenstein-saccharin solid form D, expressed as a diffraction angle of 2θ±0.2°, shows diffraction peaks at 9.20°, 10.06°, and 20.85°; furthermore, the X-ray powder diffraction pattern of encefenstein-saccharin solid form D, expressed as a diffraction angle of 2θ±0.2°, also shows diffraction peaks at 13.76°, 23.66°, and 25.69°; even further... Furthermore, the X-ray powder diffraction pattern of the encefenstein-saccharin solid form D, expressed as a diffraction angle of 2θ±0.2°, also shows diffraction peaks at 6.38°, 11.09°, 14.60°, 18.03°, 18.46°, and 24.45°; furthermore, the encefenstein-saccharin solid form D has an XRPD pattern basically as shown in Figure 14; further still, the encefenstein-saccharin solid form D has one or more of the diffraction peaks listed in Table A-5.

[0018] In some embodiments, the encefenstein-saccharin solid form D has one or more of the following characteristics:

[0019] (1) The TGA curve of ensifine-saccharin solid form D showed a weight loss of 5-10% at 224.54±3℃, for example, a weight loss of 5.15%;

[0020] (2) The DSC curve of ensefenine-saccharin solid form D shows that melting occurs at the onset point of 224.54±3℃, and the peak melting temperature is 227.55±3℃.

[0021] According to an embodiment of the present invention, the solid form comprises a eutectic or salt of the compound shown in Formula I.

[0022] On the other hand, the present invention also provides a method for preparing the solid form of encefentin-saccharin, characterized in that the solid form of encefentin-saccharin is prepared by wet milling.

[0023] The preparation method includes the following steps:

[0024] a) Mix encefentin and saccharin in a molar ratio of 1:0.9 to 1:1.2 (preferably, mix in equimolar amounts);

[0025] b) Add solvent to the mixture obtained in the previous step and grind. Preferably, the solvent is selected from acetone, ethanol, and N,N-dimethylformamide.

[0026] Preferably, the mass-to-volume ratio of encefentin to the solvent is 3 mg:(1.5–2.5) μL.

[0027] Preferably, the grinding can be performed using a vibratory grinder or a planetary ball mill.

[0028] In some embodiments, the grinding time is 1.5 to 2.5 hours (e.g., 2 hours). In some embodiments, solid form A can be obtained by grinding with a vibratory mill for 1.5 to 2.5 hours;

[0029] In some embodiments, the grinding time is 2.5-3.5 hours (e.g., 3 hours). In some embodiments, solid form B can be obtained by grinding with a planetary ball mill for 2.5-3.5 hours;

[0030] In some embodiments, the grinding time is 3.5-4.5 hours (e.g., 4 hours). In some embodiments, a solid form D can be obtained by grinding with a planetary ball mill for 3.5-4.5 hours;

[0031] In some embodiments, after obtaining solid form B, solid form C can be obtained by further steps as follows: placing solid form B in a reaction vessel and adding 15-25 times the volume (e.g., 20 times the volume) of water to suspend it for 4.5-5.5 hours (e.g., 5 hours).

[0032] In some embodiments, after obtaining solid form A, solid form B can be obtained by further steps as follows: placing solid form A in a reaction vessel and adding 15-25 times the volume (e.g., 20 times the volume) of ethanol to suspend it for 4.5-5.5 hours (e.g., 5 hours).

[0033] In some embodiments, after obtaining solid form A, solid form C can be obtained by further steps as follows: placing solid form A in a reaction vessel and adding 15-25 times the volume (e.g., 20 times the volume) of water to suspend it for 4.5-5.5 hours (e.g., 5 hours).

[0034] In some embodiments, in step b), a planetary ball mill is used to grind the material for 4.5-5.5 hours (e.g., 5 hours) to obtain a co-amorphous form; thereafter, the co-amorphous form is placed in a reaction vessel and suspended in water at 15-25 times the volume of the solids (e.g., 20 times the volume) for 4.5-5.5 hours (e.g., 5 hours) to obtain encefentin-saccharin solid form C.

[0035] In some embodiments, in step b), a planetary ball mill is used to grind the material for 4.5-5.5 hours (e.g., 5 hours) to obtain a co-amorphous form; thereafter, the co-amorphous form is placed in a reaction vessel and suspended in 15-25 times the volume (e.g., 20 times the volume) of ethanol for 4.5-5.5 hours (e.g., 5 hours) to obtain encefentin-saccharin solid form B.

[0036] In some embodiments, after obtaining solid forms B and C, solid form A can be further obtained by heating solid forms B and C to above 50°C. In another aspect, the present invention provides a pharmaceutical composition comprising one or more of the encefentin-saccharin solid forms. According to embodiments of the present invention, the encefentin-saccharin solid form in the pharmaceutical composition is a therapeutically effective amount.

[0037] In some embodiments, the dosage form of the pharmaceutical composition is selected from, but is not limited to, suspensions and dry powder inhalers for nebulized inhalation.

[0038] In another aspect, the present invention provides the use of the encefentin-saccharin solid form or the pharmaceutical composition thereof in the preparation of a medicament for the prevention and / or treatment of adult chronic obstructive pulmonary disease, noncystic bronchiectasis, asthma, cystic fibrosis, allergic rhinitis, etc.

[0039] According to an embodiment of the present invention, the encefentin-saccharin solid form can be used in combination with one or more other active therapeutic agents.

[0040] The present invention also relates to methods for preventing and / or treating diseases such as chronic obstructive pulmonary disease, noncystic bronchiectasis, asthma, cystic fibrosis, and allergic rhinitis in adults, said methods comprising administering to a patient a therapeutically effective amount of the encefentin-saccharin solid form or the pharmaceutical composition thereof.

[0041] Terminology Definitions and Explanations

[0042] Unless otherwise stated, the definitions of groups and terms recorded in this application specification and claims, including their definitions as examples, exemplary definitions, preferred definitions, definitions recorded in tables, and definitions of specific compounds in the examples, can be arbitrarily combined and combined with each other. Such combinations and combinations of group definitions and compound structures should fall within the scope of this application specification.

[0043] The term "patient" refers to any animal, including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, pigs, cattle, sheep, horses, or primates, with humans being the most preferred. For example, patients include those with chronic obstructive pulmonary disease, noncystic bronchiectasis, asthma, cystic fibrosis, allergic rhinitis, and other similar conditions.

[0044] The term "therapeutic effective dose" refers to the range of effective doses commonly used in clinical practice. It lies between the minimum effective dose and the maximum effective dose, achieving both good therapeutic effect and relative safety.

[0045] The term "pharmaceutical acceptable" refers to a set of standards and conditions that a drug must meet in its research, development, and clinical application to ensure its safety and efficacy.

[0046] The term "encefentin-saccharin solid form" refers to a crystal containing encefentin and saccharin bonded together in the lattice through non-covalent interactions. The solid form can be a eutectic or a salt, and can encompass both non-hydrated and hydrated forms. The term "X-ray powder diffraction pattern substantially as shown" means that 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 the major peaks shown in the X-ray powder diffraction pattern appear in the pattern; the major peaks refer to peaks with a relative intensity greater than 10%, preferably greater than 20%, and more preferably greater than 30%, with the highest peak as a reference (the relative intensity of the highest peak is specified as 100%).

[0047] The term "X-ray powder diffraction" or "XRPD" refers to a structural analysis method that uses X-rays to irradiate crystalline powder and utilizes the X-ray diffraction formed by the crystalline powder to analyze the spatial distribution of atoms within a substance.

[0048] The term "scanning differential calorimetry" or "DSC" refers to a thermal analysis method that measures the change in energy difference between a test sample and a reference under programmed temperature conditions. Unless otherwise specified, the reference in this article refers to an empty sample tray of the same model as the one used to load the test sample.

[0049] The term "thermogravimetric analysis" or "TGA" refers to a thermal analysis method that measures the change in sample mass with ambient temperature under programmed temperature conditions.

[0050] The term "high performance liquid chromatography" or "HPLC" refers to the method and technique of separating each component in a quantitative mixture using the principles of liquid chromatography.

[0051] The term "nuclear magnetic resonance" or "NMR" refers to the analysis of the composition and structure of matter by studying the absorption of radio frequency radiation by atomic nuclei.

[0052] The term "nuclear magnetic resonance hydrogen spectrum" or " 1 "H NMR" refers to the method and technique of using the response of hydrogen nuclei in a magnetic field to determine the position and number of hydrogen atoms on the carbon skeleton of organic molecules, thereby inferring the carbon skeleton structure of organic compounds.

[0053] The term "orthogonally polarized light microscopy" or "PLM" refers to the microscopic imaging of a material under orthogonally polarized light sources. Generally speaking, crystalline materials emit light under orthogonally polarized light, while amorphous materials do not.

[0054] The term "API" refers to the free state of encefenstein. Beneficial effects

[0055] Compared with the prior art, the present invention has the following advantages:

[0056] 1) The solid form of encefentin-saccharin of the present invention, compared with the free crystal form, can effectively improve the solubility of encefentin (and thus improve bioavailability), and has good physicochemical stability, and can be widely used in the development of better formulations;

[0057] 2) In addition to nebulized inhalers and dry powder inhalers, the solid form of the present invention, saccharin-saccharin, can be widely used in other drug dosage forms. Attached Figure Description

[0058] Figure 1 shows the XRPD diagram of Ensefentin-saccharin solid form A;

[0059] Figure 2 shows the XRPD comparison of Ensefentin-saccharin solid form A, API after grinding, saccharin after grinding, and their physical mixture;

[0060] Figure 3 shows the TGA and DSC overlay images of Ensefin-Saccharin solid form A;

[0061] Figure 4 shows the solid form A of enfleurone-saccharin. 1 H NMR spectrum;

[0062] Figure 5 shows the PLM diagram of Ensefentine-saccharin solid form A;

[0063] Figure 6 shows API (bottom), Enfertin-saccharin solid form A (middle), and saccharin (top). 1 H NMR overlay;

[0064] Figure 7 shows the XRPD pattern of Ensefentin-saccharin solid form B;

[0065] Figure 8 shows a comparison of the solid form of ensefentin-saccharin (B), XRPD after grinding saccharin, and XRPD after grinding API.

[0066] Figure 9 shows the XRPD pattern of Ensefentin-saccharin solid form B after heating and crystallization into solid form A.

[0067] Figure 10 shows the solid form of enfleurone-saccharin B. 1 H NMR spectrum;

[0068] Figure 11 shows the XRPD pattern of Ensefentin-saccharin solid form C;

[0069] Figure 12 shows the XRPD pattern of solid form A obtained after drying of solid form C of ensefenine-saccharin;

[0070] Figure 13 shows the TGA and DSC spectra of Ensefentin-saccharin solid form A after heating and drying;

[0071] Figure 14 shows the XRPD diagram of the solid form D of ensefenine-saccharin;

[0072] Figure 15 shows the TGA and DSC overlay images of Ensefentin-saccharin solid form D;

[0073] Figure 16 is a PLM diagram of Ensefin-Saccharin solid form D;

[0074] Figure 17 shows the ORTEP diagram of Ensefentine-saccharin solid form A; Detailed Implementation

[0075] The technical solution of the present invention will be further described in detail below with reference to specific embodiments. It should be understood that the following embodiments are merely illustrative and explanatory of the present invention, and should not be construed as limiting the scope of protection of the present invention. All technologies implemented based on the above content of the present invention are covered within the scope of protection intended by the present invention.

[0076] Unless otherwise stated, the raw materials and reagents used in the following examples are commercially available products or can be prepared by known methods.

[0077] I. The instruments and testing methods used in this invention are as follows:

[0078] 1.1 X-ray powder diffraction (XRPD)

[0079] 1.2 TGA Detection Method

[0080] 1.3 DSC Detection Method

[0081] 1.4 HPLC Detection Method

[0082] 1.5 X-ray single crystal diffraction

[0083] II. Reagent Abbreviation Reference Table

[0084] Table of Chinese and English names of solvents used in the experiment

[0085] Unless otherwise stated, the "API" or "encefentin raw material" used in the embodiments of the present invention refer to the encefentin free state compound, which can be purchased from Shanghai Bohou Biotechnology Co., Ltd. or synthesized according to the method disclosed in patent CN100415743C. Its crystal form is the encefentin free state crystal form I reported in the literature (Ananya Kar, et al., CrystEngComm, 2024, 26, 3783).

[0086] Example 1: Preparation of Encerfentin in solid form

[0087] The experimental steps are as follows:

[0088] Approximately 100 mg of sample and the corresponding ligand were weighed equimolarly into the grinding jar of a vibratory grinder. Ethanol or water was added as a solvent. The vibration frequency of the vibratory grinder was set to approximately 400 Hz. After ball milling for 2 hours, the solid material was vacuum dried at 50°C. The obtained sample was subjected to XRPD, TGA, DSC, and other analytical techniques. 1 Characterization by HNMR and other methods, the results are shown in Table 1. Only saccharin and encefentin formed a new solid form.

[0089] Table 1

[0090] Example 2: Preparation of Encerfentin-Saccharin Solid Form

[0091] a) Weigh 20 mg of API sample into a centrifuge tube and mix with an appropriate amount of saccharin at a molar ratio of 1:1.

[0092] b) Add 15 μL of EtOH and place it in a vibratory mill. After milling for 2 hours, solid form A is obtained.

[0093] Crystal form identification results: XRPD results (Figure 1) show that encefentin-saccharin solid form A has good crystallinity, and the X-ray diffraction peak positions of encefentin-saccharin solid form A are different from those of encefentin free state and saccharin (Figure 2). TGA / DSC results are shown in Figure 3. TGA results show that the sample loses 3.34% weight before 97.12℃; DSC results show that the sample has a single melting point. The TGA / DSC results after heating and drying solid form A are shown in Figure 13. TGA results show that the sample loses less weight before the melting point; DSC results show that the sample has a single melting point. 11H NMR showed that the peak positions of the 1H NMR spectra of the solid forms of encefentin and saccharin were shifted compared to those of the free forms (Figures 4 and 6). Polarizing microscopy revealed that solid form A consisted of bulk crystalline particles (Figure 5). The crystallographic parameters of the encefentin-saccharin solid form A prepared in this invention are (Table A-2): chiral space group P21 / n; cell parameters are: α=90°, β=91.662(3)°, γ=90°, The number of molecules (Z) in a unit lattice is 4, and the density (D) is 1.374 g / cm³. 3 The ORTEP plot of the encefentin-saccharin solid form A of the present invention shows that one molecule of encefentin combines with one molecule of saccharin to form a solid form, which is solvent-free, as shown in Figure 17. The XRPD analysis data of the encefentin-saccharin solid form A are shown in Table A-1 below:

[0094] Table A-1 XRPD diffraction peak data of Ensefinitin-Saccharin solid form A

[0095] Table A-2 Main crystallographic data of Enfertin-Saccharin solid form A

[0096] Example 3: Preparation of Encerfentin-Saccharin Solid Form

[0097] a) Weigh 300 mg of API sample into a grinding jar, mix with an appropriate amount of saccharin at a molar ratio of 1:1, and add 300 μL of EtOH;

[0098] b) The grinding jar was placed in a planetary ball mill and ground for 4 hours to obtain enfrenzyme-saccharin solid form B (Figure 7). The diffraction peak positions of solid form B are different from those of enfrenzyme free state and saccharin alone (Figure 8). Heating experiments on solid form B revealed that solid form B can be converted into solid form A after heating (Figure 9). 1 Figure 10 shows the 1H NMR spectra of solids A, B, and C in solution. 1 (The H NMR values ​​are consistent);

[0099] c) Place Ensefentin-saccharin solid form B in a glass reaction flask, add water at 20 times the volume of the solid, stir for 5 hours to obtain Ensefentin-saccharin solid form C (wet product, Figure 11), and dry to obtain Ensefentin-saccharin solid form A (Figure 12).

[0100] The above experiments show that encefentin-saccharin in solid form is suspended in ethanol solution as solid form B. After losing ethanol, the solid form can quickly transform into solid form A. Therefore, it is particularly suitable for the preparation of materials in ethanol system. Encefentin-saccharin in aqueous solution is suspended in solid form C. After losing water, the solid form can quickly transform into solid form A. Therefore, solid form C is particularly suitable for the development of aqueous suspension formulations.

[0101] Example 4: Preparation of Encerfentin-Saccharin Solid Form

[0102] a) Weigh 300 mg of API sample into a grinding jar, mix with an appropriate amount of saccharin at a molar ratio of 1:1, and add 300 μL of EtOH;

[0103] b) Place the grinding jar in a planetary ball mill and grind for 5 hours to obtain amorphous material;

[0104] c) The amorphous material was placed in a glass reaction flask, and water with a volume 20 times the solid amount was added and the mixture was suspended for 5 hours to obtain Ensefentin-saccharin solid form C (wet product). After drying, Ensefentin-saccharin solid form A was obtained.

[0105] Example 5: Preparation of Encerfentin-Saccharin Solid Form

[0106] a) Weigh 300 mg of API sample into a grinding jar, mix with an appropriate amount of saccharin at a molar ratio of 1:1, and add 300 μL of EtOH;

[0107] b) Place the grinding jar in a planetary ball mill and grind for 5 hours to obtain amorphous material;

[0108] c) The amorphous material was placed in a glass reaction flask, and EtOH with a volume of 20 times the solid amount was added and the mixture was suspended for 5 hours to obtain Ensefentine-saccharin solid form B (wet product). After drying, Ensefentine-saccharin solid form A was obtained.

[0109] The XRPD analysis data of the solid forms B and C of ensefenine-saccharin obtained above are shown in Tables A-3 and A-4 below, respectively:

[0110] Table A-3 XRPD diffraction peak data of Ensefinitin-Saccharin solid form B

[0111] Table A-4 XRPD diffraction peak data of Ensefinitin-Saccharin solid form C

[0112] Example 6: Preparation of Encerfentin-Saccharin Solid Form

[0113] a) Weigh 4g of API sample into a grinding jar, add 1.5g of saccharin at a molar ratio of 1:1 and mix well;

[0114] b) Add 4 mL of EtOH; place the grinding jar in a planetary ball mill and grind for 4 hours to obtain ensefenine-saccharin solid form D.

[0115] Crystal form identification results: XRPD results (Figure 14) show that encefentin-saccharin solid form D has good crystallinity. TGA / DSC results are shown in Figure 15. TGA results show that the sample lost 5.15% weight before reaching 224.54℃; DSC results show that the sample has a single melting point. Polarizing microscopy reveals that the crystal morphology of solid form D is irregular fragmented (Figure 16). The XRPD analysis data for encefentin-saccharin solid form D are shown in the table below:

[0116] Table A-5 XRPD diffraction peak data of Ensefinitin-Saccharin solid form D

[0117] Example 7: Solubility test of the solid form of the present invention

[0118] The saturated solubility (37°C) of the free crystal form I and the encefentin-saccharin solid form in water was measured, and the results are shown in the table below. The solid form of the present invention has a significantly improved solubility compared to the free crystal form.

[0119] Example 8: Stability test of enstatin-saccharin solid form A

[0120] The experimental results above show that Ensefentin-saccharin solid form A can exist stably in a solid state under low humidity conditions, making solid form A more advantageous for the development of dry powder inhalers; solid form B is stable under higher humidity conditions and in ethanol suspension systems, making solid form B more suitable for the development of ordinary oral solid dosage forms (such as tablets) or the preparation of materials in ethanol systems; solid form C can exist stably in an aqueous suspension system, making solid form C more advantageous for the development of suspensions for nebulized inhalation.

[0121] Example 9: Stability Study of Encerfentin-Saccharin Solid Suspension

[0122] Take 50 mg each of solid form B and solid form C, add 20 times the volume of solvent by weight of the solid at 25 °C to suspend them, and test the stability of the solid forms at 7 days and 14 days.

[0123] Example 10: Stability test of enfleurone-saccharin solid form D

[0124] The stability of the solid form of encefenidine-saccharin in solid form D was investigated, and the experimental results are as follows:

[0125] Example 11: Determination of the solubility curve of encefentanil-saccharin solid form D as a function of pH

[0126] Weigh approximately 20 mg of sample into a 1.5 mL glass vial; add 1 mL of different pH buffer solutions to the vial to form a suspension; place the resulting suspension on a shaker at 37°C (150 rpm); after 24 hours, take 100 μL of the supernatant, dilute it with 400 μL of methanol, send it for testing, and test the final pH of the suspension. The solubility of each solid form at different pH values ​​is obtained, as shown in the table below.

[0127] Example 12 Competitive interconversion relationship between encefentin-saccharin solid form A and encefentin-saccharin solid form D

[0128] 50 mg of each of encefentin-saccharin solid form A and encefentin-saccharin solid form D were weighed into 1.5 mL glass vials. 1 mL of different solvents (water and ethanol) were added to the vials to prepare aqueous suspensions of form A and D and ethanol suspensions of form A and D. After stirring for four days, the solids were separated and detected. Encefentin-saccharin solid form D was obtained in both different suspension systems (water and ethanol).

[0129] The embodiments of the present invention have been described above. However, the present invention is not limited to the above embodiments. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. An encefenstein-saccharin solid form having the structure shown in Formula I:

2. The encefentin-saccharin solid form according to claim 1, characterized in that, The solid form is encefenstein-saccharin solid form A. The X-ray powder diffraction pattern of encefenstein-saccharin solid form A, expressed as a diffraction angle of 2θ±0.2°, shows diffraction peaks at 10.29°, 11.12°, and 19.81°. Furthermore, the X-ray powder diffraction pattern of encefenstein-saccharin solid form A, expressed as a diffraction angle of 2θ±0.2°, also shows diffraction peaks at one or more of the following locations: 8.34°, 10.89°, 21.92°, 22.45°, and 23.61°. Even further... Furthermore, the X-ray powder diffraction pattern of the encefentin-saccharin solid form A, expressed as a diffraction angle of 2θ±0.2°, also has diffraction peaks at one or more of the following locations: 11.69°, 13.30°, 15.03°, 15.50°, 19.10°, 25.59°, and 28.38°; furthermore, the encefentin-saccharin solid form A has an XRPD pattern basically as shown in Figure 1; further still, the encefentin-saccharin solid form A has one or more of the diffraction peaks listed in Table A-1.

3. The encefentin-saccharin solid form according to claim 2, characterized in that, The encefenidine-saccharin solid form A has one or more of the following characteristics: (1) The TGA curve of ensifine-saccharin solid form A showed a weight loss of 0-5% at 180.39±5℃, for example, a weight loss of 3.34%; (2) The DSC curve of Ensefentin-saccharin solid form A shows that melting occurs at the onset point of 209.29±5℃, and the peak melting temperature is 212.77±5℃. Preferably, the TGA / DSC chart of the encefenstein-saccharin solid form A is basically as shown in Figure 3; the encefenstein-saccharin solid form A... 1 The basic H NMR spectrum is shown in Figure 4.

4. The encefentin-saccharin solid form A according to claim 1, characterized in that, Its crystallographic parameters are: chiral space group P2I / n; unit cell parameters are: α=90°, β=91.662(3)°, γ=90°, The number of molecules (Z) in a unit lattice is 4, and the density (D) is 1.374 g / cm³. 3 Furthermore, the crystallographic parameters of the encefentin-saccharin solid morphology A are shown in Table A-2.

5. The encefentin-saccharin solid form according to claim 1, characterized in that, The solid form is encefenidine-saccharin solid form B. The X-ray powder diffraction pattern of encefenidine-saccharin solid form B, expressed as a diffraction angle of 2θ±0.2°, shows diffraction peaks at 10.32°, 11.12°, and 19.58°. Furthermore, the X-ray powder diffraction pattern of encefenidine-saccharin solid form B, expressed as a diffraction angle of 2θ±0.2°, also shows diffraction peaks at one or more of 8.19°, 15.13°, and 23.23°. Even further, the encefenidine-saccharin solid form B... The X-ray powder diffraction pattern of solid form B, expressed as a diffraction angle of 2θ±0.2°, also shows diffraction peaks at one or more of the following locations: 11.69°, 17.00°, 18.62°, 20.97°, 21.69°, 22.34°, 25.09°, and 25.50°. Furthermore, the encefentin-saccharin solid form B has an XRPD pattern basically as shown in Figure 7. Even further, the encefentin-saccharin solid form B has one or more of the diffraction peaks listed in Table A-3.

6. The encefentin-saccharin solid form according to claim 1, characterized in that, The solid form is encefenstein-saccharin solid form C. The X-ray powder diffraction pattern of encefenstein-saccharin solid form C, expressed as a diffraction angle of 2θ±0.2°, shows diffraction peaks at 15.14°, 19.36°, and 22.96°. Furthermore, the X-ray powder diffraction pattern of encefenstein-saccharin solid form C, expressed as a diffraction angle of 2θ±0.2°, also shows diffraction peaks at one or more of 10.29°, 11.08°, and 22.43°. Even further, the X-ray powder diffraction pattern of encefenstein-saccharin solid form C, expressed as a diffraction angle of 2θ±0.2°, also shows a peak at 8.07°. The encefenstein-saccharin solid form C exhibits diffraction peaks at one or more of the following locations: 13.14°, 18.67°, and 22.18°. Furthermore, the X-ray powder diffraction pattern of the encefenstein-saccharin solid form C, expressed as a diffraction angle of 2θ ± 0.2°, also exhibits diffraction peaks at one or more of the following locations: 11.65°, 19.70°, 20.95°, 21.17°, 21.50°, 25.07°, and 25.41°. Further, the encefenstein-saccharin solid form C has an XRPD pattern essentially as shown in Figure 11. Even further, the encefenstein-saccharin solid form C has one or more diffraction peaks listed in Table A-4.

7. The encefentin-saccharin solid form according to claim 1, characterized in that, The solid form is encefenstein-saccharin solid form D. The X-ray powder diffraction pattern of encefenstein-saccharin solid form D, expressed as a diffraction angle of 2θ±0.2°, shows diffraction peaks at 9.20°, 10.06°, and 20.85°. Furthermore, the X-ray powder diffraction pattern of encefenstein-saccharin solid form D, expressed as a diffraction angle of 2θ±0.2°, also shows diffraction peaks at 13.76°, 23.66°, and 25.69°. Even further, the... The X-ray powder diffraction pattern of the encefenstein-saccharin solid form D, expressed as a diffraction angle of 2θ±0.2°, also shows diffraction peaks at 6.38°, 11.09°, 14.60°, 18.03°, 18.46°, and 24.45°; furthermore, the encefenstein-saccharin solid form D has an XRPD pattern basically as shown in Figure 14; further still, the encefenstein-saccharin solid form D has one or more of the diffraction peaks listed in Table A-5; Preferably, the encefenstein-saccharin solid form D has one or more of the following characteristics: (1) The TGA curve of ensifine-saccharin solid form D showed a weight loss of 5-10% at 224.54±3℃, for example, a weight loss of 5.15%; (2) The DSC curve of ensefenine-saccharin solid form D shows that melting occurs at the onset point of 224.54±3℃, and the peak melting temperature is 227.55±3℃. Preferably, the TGA / DSC plot of the encefentin-saccharin solid form D is basically as shown in Figure 15.

8. The method for preparing encerfentin-saccharin in solid form according to any one of claims 1-7, characterized in that, The preparation method includes the following steps: a) Mix encefentin and saccharin in a molar ratio of 1:0.9 to 1:1.2 (preferably, mix in equimolar amounts); b) Add solvent to the mixture obtained in the previous step and grind; preferably, the solvent is selected from acetone, ethanol and N,N-dimethylformamide; Preferably, the mass-to-volume ratio of encefentin to the solvent is 3 mg:(1.5–2.5) μL; Preferably, the grinding can be performed using a vibratory grinder or a planetary ball mill; Preferably, the mass-to-volume ratio of solid encefentin to the solvent is 3 mg:(1.5–2.5) μL; Preferably, the grinding can be performed using a vibratory grinder or a planetary ball mill; Preferably, solid form A can be obtained by grinding with a vibratory grinder for 1.5 to 2.5 hours; Preferably, solid form B can be obtained by grinding with a planetary ball mill for 2.5-3.5 hours; Preferably, solid form D can be obtained by grinding with a planetary ball mill for 3.5-4.5 hours; Preferably, after obtaining solid form B, the following steps can be used to obtain solid form C: place solid form B in a reaction vessel and add 15-25 times the volume of water to suspend it for 4.5-5.5 hours; Preferably, after obtaining solid form A, solid form B can be obtained by further adopting the following steps: placing solid form A in a reaction vessel and adding 15-25 times the volume of solid ethanol for 4.5-5.5 hours; Preferably, after obtaining solid form A, the following steps can be used to obtain solid form C: place solid form A in a reaction vessel and add 15-25 times the volume of water to suspend it for 4.5-5.5 hours; Preferably, in step b), a planetary ball mill is used to grind the material for 4.5-5.5 hours to obtain a co-amorphous form; thereafter, the co-amorphous form is placed in a reaction vessel, and water with a volume of 15-25 times the solid amount is added and the mixture is suspended for 4.5-5.5 hours to obtain encefentin-saccharin solid form C. Preferably, in step b), a planetary ball mill is used to grind the material for 4.5-5.5 hours to obtain a co-amorphous form; thereafter, the co-amorphous form is placed in a reaction vessel and suspended in ethanol at 15-25 times its volume of solids for 4.5-5.5 hours to obtain encefentin-saccharin solid form B. Preferably, after obtaining solid forms B and C, the following steps can be used to obtain solid form A: heating solid forms B and C to above 50°C to obtain solid form A.

9. A composition comprising one or more of the ensifine-saccharin solid forms according to any one of claims 1-7; Preferably, the dosage form of the pharmaceutical composition is selected from nebulized suspensions and dry powder inhalers; preferably, the encefentin-saccharin solid form can be used in combination with one or more other active therapeutic drugs.

10. Use of the encefentin-saccharin solid form according to any one of claims 1-8 in the preparation of a medicament, wherein the medicament is used for the prevention and / or treatment of adult chronic obstructive pulmonary disease, noncystic bronchiectasis, asthma, cystic fibrosis, allergic rhinitis, etc.

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