Nylestriol crystalline form c and process for its preparation

Abstract

The present application relates to a crystalline form C of nylestriol and a preparation method thereof, and belongs to the technical field of biological medicine. The DSC spectrum of the crystalline form C shows endothermic melting at 167.16 DEG C and reaches a melting peak at 168.85 DEG C. The preparation method comprises the following steps: dissolving nylestriol in dichloromethane or other solvents, heating to complete dissolution, cooling and crystallizing at a stirring speed of 200-350 rpm, incubating at 10-45 DEG C for 1-8 hours, filtering and drying to obtain the crystalline form C. The purity of the crystalline form C prepared by the present application is more than 99.4%, the stability is good, the preparation condition is mild, the operation is simple, the present application is suitable for industrial production, and the present application can be used for preparing long-acting estrogen drugs.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, specifically a method for preparing somatic estradiol crystal form C. Background Technology

[0002] During the perimenopausal and postmenopausal periods, women experience a continuous decline in estrogen levels due to ovarian dysfunction, which can easily lead to a series of menopausal-related syndromes, such as hot flashes, night sweats, insomnia, irritability, reproductive tract atrophy, osteoporosis, and cardiovascular and metabolic disorders, severely impacting their quality of life. Estrogen replacement therapy is the core clinical intervention for these conditions; however, traditional estrogen-based drugs have drawbacks such as excessively strong biological activity, short duration of action, high frequency of use, significant endometrial irritation, and poor long-term safety. Clinically, there is an urgent need for a long-acting, gentle, safe, and well-adhered-to-use estrogen replacement drug.

[0003] Ethylestradiol, a commonly used long-acting estrogen drug in clinical practice, is a synthetic derivative of estriol. Estriol itself is a natural metabolite of estradiol in the body. Its pharmacological effects are highly similar to those of estradiol, but its core advantage lies in its lower biological activity and weaker stimulation of endometrial proliferation. It is suitable for the clinical needs of perimenopausal women for long-term, gentle estrogen supplementation. Compared with potent estrogens such as estradiol and ethinylestradiol, it has a significant safety advantage and is one of the preferred drugs for postmenopausal estrogen replacement therapy.

[0004] Nereestrol's chemical name is 3-(cyclopentyloxy)-19-demethyl-17α-pregn-1,3,5(10)-trien-20-yn-16α,17α-diol, and its molecular structure is as follows: ; The molecular structure of estriol is specifically modified to possess unique pharmacokinetic advantages: the introduction of a cyclopentyl ether group at the 3 position of the parent estriol structure significantly enhances the drug's lipophilicity, making it easier for the intestines to absorb and allowing it to slowly accumulate and be continuously released in adipose tissue, achieving a long-lasting pharmacological effect; at the same time, the introduction of an ethynyl group at the 17 position moderately enhances the estrogen receptor binding activity, making up for the shortcoming of the low natural activity of estriol, and balancing the mildness of the drug with the effectiveness of its action.

[0005] At the pharmacological level, norestrol can specifically bind to estrogen receptors in the body, effectively supplementing estrogen deficiency in postmenopausal women, alleviating various perimenopausal discomfort symptoms, improving reproductive tract atrophy problems such as senile vaginitis and atrophic urethritis, while also inhibiting bone resorption, promoting bone formation, preventing postmenopausal osteoporosis, regulating lipid metabolism, and reducing the risk of postmenopausal cardiovascular disease. Because of its low irritant effect on the endometrium, it does not require routine use with progesterone antagonists, making clinical medication regimens simpler and applicable to a wider range of populations.

[0006] Currently, there are significant gaps in the systematic screening, preparation, characterization, and quality control of sonyestradiol crystal forms in existing publicly available technologies, pharmacopoeia standards, and commercially available sonyestradiol preparations both domestically and internationally. Existing industrial production processes also exhibit significant crystal form-related defects.

[0007] Therefore, conducting specialized research and development on the crystal form of norethisterone to obtain a stable, high-quality, and industrially producible new crystal form, and establishing supporting preparation and quality control methods, can not only break through existing technological bottlenecks and improve the overall quality of norethisterone raw materials and preparations, meeting the clinical demand for high-quality, highly stable estrogen replacement drugs, but also form a core technology patent barrier, fill the gaps in existing technology and patent layout, and have important pharmaceutical research value, clinical application value, and market economic benefits. This is also the core technological starting point and basis for the establishment of this patent application. Summary of the Invention

[0008] To solve the above problems, the technical solution provided by the present invention is as follows: The crystalline form C of nystatin of the present invention, when subjected to Cu-Kα radiation, exhibits characteristic peaks in its 2θ X-ray powder diffraction pattern at the following positions: 3.4±0.2°, 6.6±0.2°, 8.2±0.2°, 10.7±0.2°, 13.7±0.2°, 14.6±0.2°, 15.0±0.2°, 15.8±0.2°, 16.6±0.2°, 1 7.0±0.2°, 17.3±0.2°, 19.0±0.2°, 19.2±0.2°, 20.3±0.2°, 20.5±0.2°, 21.4±0.2°, 21.6±0.2°, 23.0±0.2°, 23.4±0.2°, 24.6±0.2°, 25.0±0.2°, 25.6±0.2°, 26.7±0.2°.

[0009] Preferably, the DSC spectrum of soliestrol in crystal form C begins to undergo endothermic melting at 167.16 °C and reaches the peak melting rate at 168.85 °C.

[0010] A process for preparing crystalline form C of soliestrol includes the following steps. (1) Dissolve solenoid estradiol in dichloromethane or other solvent and heat until completely dissolved; (2) Stir and cool the solution obtained in step (1) until the material precipitates; (3) Keep the rotation speed and keep the material in step (2) warm until all the solid precipitates out. Then filter, collect the filter cake, and dry it to obtain crystalline form C of solenoid estradiol.

[0011] Preferably, the other solvents in step (1) include at least one of haloalkanes, alcohols, ketones, and esters.

[0012] Preferably, the halogenated hydrocarbon solvent includes at least one of dichloromethane, trichloromethane, and dichloroethane; the alcohol solvent includes at least one of methanol, ethanol, and isopropanol; the ketone solvent includes at least one of acetone, butanone, and cyclohexanone; and the ester solvent includes at least one of ethyl acetate and methyl acetate.

[0013] Preferably, in step (1), the mass-to-volume ratio of estradiol to dichloromethane or other solvents is 1:1 to 1:15.

[0014] Preferably, the dissolution temperature in step (1) is 50~80℃.

[0015] Preferably, the stirring speed in step (2) is in the range of 200 rpm to 350 rpm.

[0016] Preferably, the heat preservation temperature in step (3) is 10~45℃ and the stirring crystallization time is 1~8 hours.

[0017] Compared with the prior art, the technical solution provided by this invention has the following advantages: This invention discloses a nystatin crystalline form C and its preparation method. The X-ray powder diffraction pattern of crystalline form C, expressed as 2θ, obtained using Cu-Kα radiation, is shown at 3.4±0.2°, 6.6±0.2°, 8.2±0.2°, 10.7±0.2°, 13.7±0.2°, 14.6±0.2°, 15.0±0.2°, 15.8±0.2°, 16.6±0.2°, 17.0±0.2°, 17.3±0.2°, and 19... Characteristic peaks are observed at 0±0.2°, 19.2±0.2°, 20.3±0.2°, 20.5±0.2°, 21.4±0.2°, 21.6±0.2°, 23.0±0.2°, 23.4±0.2°, 24.6±0.2°, 25.0±0.2°, 25.6±0.2°, and 26.7±0.2°. The DSC spectrum shows endothermic melting at 167.16℃ and reaches the melting peak at 168.85℃. The preparation method includes: dissolving solenoid estradiol in dichloromethane or other solvent, heating until completely dissolved, cooling to crystallize while stirring at 200–350 rpm, maintaining the temperature at 10–45℃ for 1–8 hours, filtering, and drying to obtain the final product. The crystal form C obtained by this invention has a purity of over 99.4%, good stability, mild preparation conditions, and simple operation, making it suitable for industrial production and applicable to the preparation of long-acting estrogen drugs. Attached Figure Description

[0018] Figure 1 HPLC chromatogram of sample of soliestrol in crystal form C Figure 2HPLC chromatogram of a sample of soliestrol (crystal form C) after storage at 40±2℃ and 70±5°C for 6 months. Figure 3 X-ray powder diffraction (XRD) pattern of nystatin C crystal form Figure 4 Differential scanning thermal analysis (DSC) plot of soliestrol in crystal form C. Detailed Implementation

[0019] To enable those skilled in the art to better understand the present application, the technical solutions in the embodiments of the present application will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present application, and not all embodiments. Based on the embodiments in the present application, all other embodiments obtained by those of ordinary skill in the art without creative effort should fall within the scope of protection of the present application.

[0020] It should be noted that the terms "first," "second," etc., in the specification, claims, and accompanying drawings of this application are used to distinguish similar objects and are not necessarily used to describe a specific order or sequence. It should be understood that such data can be interchanged where appropriate for the embodiments of this application described herein. Furthermore, the terms "comprising" and "having," and any variations thereof, are intended to cover non-exclusive inclusion; for example, a process, method, system, product, or apparatus that comprises a series of steps or units is not necessarily limited to those steps or units explicitly listed, but may include other steps or units not explicitly listed or inherent to such processes, methods, products, or apparatus.

[0021] It should be noted that, unless otherwise specified, the embodiments and features described in this application can be combined with each other. This application will now be described in detail with reference to the accompanying drawings and embodiments.

[0022] Example 1 (1) Dissolve 5g of sori estradiol in 30ml of dichloromethane and heat to 50℃ until completely dissolved; (2) Stir the solution obtained in step (1) at a stirring speed of 200~210 rpm to cool it down until the material precipitates; (3) Keep the rotation speed and keep the material in step (2) at a temperature of 20~25℃ for 2 hours until all the solid is precipitated. Then filter, collect the filter cake, and dry it to obtain crystalline form C of solenoid estradiol.

[0023] The HPLC chromatogram of nystatin C was obtained by detection as follows: Figure 1 As shown, its purity is 99.45%.

[0024] The HPLC chromatogram of the sample after being stored at 40±2℃ and 70±5°C for 6 months is shown below. Figure 2 As shown, its purity is 99.40%, and the sample purity remains essentially unchanged.

[0025] The X-ray powder diffraction (XRD) pattern of crystalline form C of solitriol was obtained as follows: Figure 1 As shown, the feature is that Cu-Kα radiation is used, at 2θ The X-ray powder diffraction pattern shown has characteristic peaks at the following positions: 3.4±0.2°, 6.6±0.2°, 8.2±0.2°, 10.7±0.2°, 13.7±0.2°, 14.6±0.2°, 15.0±0.2°, 15.8±0.2°, 16.6±0.2°, 17.0±0.2°, 17.3±0.2°, 19.0±0.2°, 19.2±0.2°, 20.3±0.2°, 20.5±0.2°, 21.4±0.2°, 21.6±0.2°, 23.0±0.2°, 23.4±0.2°, 24.6±0.2°, 25.0±0.2°, 25.6±0.2°, 26.7±0.2°.

[0026] The differential scanning calorimetry (DSC) spectrum of nystatin C was obtained as follows: Figure 4 As shown, endothermic melting begins at 167.16 ℃ and reaches its peak melting rate at 168.85 ℃. The melting peak is sharp, symmetrical, and without tailing. The difference between the initial temperature and the peak temperature is only 1.69 ℃, indicating that the sample has a single crystal form, high crystallinity, uniform crystal structure, and excellent crystal purity.

[0027] Example 2 (1) Dissolve 5g of estradiol in 40ml of acetone and heat to 60℃ until completely dissolved; (2) Stir the solution obtained in step (1) at a stirring speed of 230~240 rpm to cool it down until the material precipitates; (3) Keep the rotation speed and keep the material in step (2) at a temperature of 15~20℃ for 3 hours until all the solid is precipitated. Then filter, collect the filter cake, and dry it to obtain crystalline form C of solenoid estradiol.

[0028] The X-ray powder diffraction pattern of the prepared crystal obtained by Cu-Kα ray measurement was the same as that in Example 1.

[0029] Example 3 (1) Dissolve 5g of sori estradiol in 35ml of ethyl acetate and heat to 55℃ until completely dissolved; (2) Stir the solution obtained in step (1) at a stirring speed of 260~270 rpm to cool it down until the material precipitates; (3) Keep the rotation speed and keep the material in step (2) at a temperature of 10~15℃ for 6 hours until all the solid is precipitated. Then filter, collect the filter cake, and dry it to obtain crystalline form C of solenoid estradiol.

[0030] The X-ray powder diffraction pattern of the prepared crystal obtained by Cu-Kα ray measurement was the same as that in Example 1.

[0031] Example 4 (1) Dissolve 5g of sori estradiol in 30ml of chloroform and heat to 55℃ until completely dissolved; (2) Stir the solution obtained in step (1) at a stirring speed of 220~230 rpm to cool it down until the material precipitates; (3) Keep the rotation speed and keep the material in step (2) at a temperature of 10~15℃ for 2 hours until all the solid is precipitated. Then filter, collect the filter cake, and dry it to obtain crystalline form C of solenoid estradiol.

[0032] The X-ray powder diffraction pattern of the prepared crystal obtained by Cu-Kα ray measurement was the same as that in Example 1.

[0033] Example 5 (1) Dissolve 5g of sori estradiol in 40ml of dichloroethane and heat to 60℃ until completely dissolved; (2) Stir the solution obtained in step (1) at a stirring speed of 240~250 rpm to cool it down until the material precipitates; (3) Keep the rotation speed and keep the material in step (2) at a temperature of 15~20℃ for 2 hours until all the solid is precipitated. Then filter, collect the filter cake, and dry it to obtain crystalline form C of solenoid estradiol.

[0034] The X-ray powder diffraction pattern of the prepared crystal obtained by Cu-Kα ray measurement was the same as that in Example 1.

[0035] Example 6 (1) Dissolve 5g of soreethrin in 40ml of methanol and heat to 50℃ until completely dissolved; (2) Stir the solution obtained in step (1) at a stirring speed of 210~220 rpm to cool it down until the material precipitates; (3) Keep the rotation speed and keep the material in step (2) at a temperature of 15~20℃ for 4 hours until all the solid is precipitated. Then filter, collect the filter cake, and dry it to obtain crystalline form C of solenoid estradiol.

[0036] The X-ray powder diffraction pattern of the prepared crystal obtained by Cu-Kα ray measurement was the same as that in Example 1.

[0037] Example 7 (1) Dissolve 5g of estradiol in 15ml of ethanol and heat to 70℃ until completely dissolved; (2) Stir the solution obtained in step (1) at a stirring speed of 300~310 rpm to cool it down until the material precipitates; (3) Keep the rotation speed and keep the material in step (2) at a temperature of 25~30℃ for 4 hours until all the solid is precipitated. Then filter, collect the filter cake, and dry it to obtain crystalline form C of solenoid estradiol.

[0038] The X-ray powder diffraction pattern of the prepared crystal obtained by Cu-Kα ray measurement was the same as that in Example 1.

[0039] Example 8 (1) Dissolve 5g of estradiol in 30ml of isopropanol and heat to 75℃ until completely dissolved; (2) Stir the solution obtained in step (1) at a stirring speed of 280~290 rpm to cool it down until the material precipitates; (3) Keep the rotation speed and keep the material in step (2) at a temperature of 30~35℃ for 4 hours until all the solid is precipitated. Then filter, collect the filter cake, and dry it to obtain crystalline form C of solenoid estradiol.

[0040] The X-ray powder diffraction pattern of the prepared crystal obtained by Cu-Kα ray measurement was the same as that in Example 1.

[0041] The detection instruments and testing methods used in this invention 1) Instrument: Bruker D2 PHASER Voltage: 30kV, Current: 10mA; Sola slit: 4.0° / 4.0°; Diverging slit: 1.0mm; Scan range: 3°-40°; Step size: 0.02°; Rate: 2° / min; Test method: Take an appropriate amount of sample on the sample tray, ensuring that the sample is flush with the sample tray, and scan according to the method.

[0042] (2) Instrument: DSC250 Test method: A 1.4 mg sample was placed in a non-sealed aluminum dish, kept at 25 °C and then heated to 350 °C at a scan rate of 10 °C / min.

[0043] In addition, in step (1), the mass-volume ratio of nystatin to one or more of the following solvents, such as halogenated hydrocarbon solvents, alcohol solvents, ketone solvents, and ester solvents, is 1:1 to 1:15, preferably 1:2 to 1:10. If the ratio is lower than 1:1, the material cannot be completely dissolved, which affects the product yield. If the ratio is higher than 1:15, the material cannot be fully crystallized, and the product yield decreases.

[0044] The dissolution temperature in step (1) is 50~80℃. If the dissolution temperature is below 50℃, the material will not be completely dissolved, affecting the product yield. If the dissolution temperature is above 80℃, some of the solvent will boil violently, posing a safety risk.

[0045] In step (2), the stirring speed range is 200 rpm to 350 rpm. If the stirring speed is lower than 200 rpm, the C-crystal form of nystatin provided by this patent cannot be obtained. If the stirring speed is higher than 350 rpm, the product obtained will have large impurities due to excessive crystallization.

[0046] The heat preservation temperature in step (3) is 10~45℃, and the stirring crystallization time is 1~8 hours. If the crystallization temperature is higher than 45℃, the material cannot be fully crystallized and the product yield will decrease.

[0047] The C-crystal form of nystatin provided by this invention has the following advantages After multiple experiments, it has been verified that the C-crystal form of nystatin provided by this invention has high purity and good yield, with a purity of over 99.4%, and good stability. After the sample was stored for 6 months at a temperature of 40±2℃ and a humidity of 70±5°, the quality of the sample did not change significantly.

[0048] Its preparation method is simple, easy to operate, and mild, requiring no special reaction conditions, thus making it suitable for industrial production.

[0049] The C-type solitriol provided in this invention differs significantly from the α-type solitriol provided in patent CN 117143167 A, because the former is naturally crystallized from one or more solvents, while the preparation method provided in this patent involves rapid stirring and crystallization after dissolution. This results in a faster material heat transfer rate and a dramatic increase in the product nucleation rate. Specifically, using Cu-Kα radiation, the X-ray powder diffraction pattern, expressed as 2θ, exhibits characteristic peaks at positions 3.4±0.2°, 6.6±0.2°, 8.2±0.2°, 10.7±0.2°, 13.7±0.2°, 14.6±0.2°, 15.0±0.2°, 16.6±0.2°, 17.0±0.2°, 19.0±0.2°, 20.3±0.2°, 20.5±0.2°, 21.4±0.2°, 23.0±0.2°, 23.4±0.2°, 24.6±0.2°, 25.6±0.2°, and 26.7±0.2°. The α peaks provided in patent CN 117143167 A... No characteristic peaks were observed at the positions described for crystalline solitriol: 12.1+0.2°, 12.8+0.2°, 13.4+0.2°, 13.9+0.2°, 15.2+0.2°, 16.4+0.2°, 17.9+0.2°, 21.1+0.2°, 22.3+0.2°, and 24.2+0.2°.

[0050] The above-described embodiments are merely illustrative of certain implementations of the present invention, and are described in a relatively specific and detailed manner. However, they should not be construed as limiting the scope of the present invention. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these modifications and improvements are all within the scope of protection of the present invention. Therefore, the scope of protection of the present invention should be determined by the appended claims.

Claims

1. A crystalline form C of soliestrol, characterized in that: Using Cu-Kα radiation, the X-ray powder diffraction pattern, expressed as 2θ, has characteristic peaks at the following positions: 3.4±0.2°, 6.6±0.2°, 8.2±0.2°, 10.7±0.2°, 13.7±0.2°, 14.6±0.2°, 15.0±0.2°, 15.8±0.2°, 16.6±0.2°, and 17.0±0.2°. °, 17.3±0.2°, 19.0±0.2°, 19.2±0.2°, 20.3±0.2°, 20.5±0.2°, 21.4±0.2°, 21.6±0.2°, 23.0±0.2°, 23.4±0.2°, 24.6±0.2°, 25.0±0.2°, 25.6±0.2°, 26.7±0.2°.

2. The crystal form C of soliestrol according to claim 1, characterized in that: The DSC spectrum of crystalline form C of solitriol shows that endothermic melting begins at 167.16 °C and reaches the peak melting rate at 168.85 °C.

3. The method for preparing crystal form C of soliestrol according to claim 1 or 2, characterized in that: Includes the following steps (1) Dissolve solenoid estradiol in dichloromethane or other solvent and heat until completely dissolved; (2) Stir and cool the solution obtained in step (1) until the material precipitates; (3) Keep the rotation speed and keep the material in step (2) warm until all the solid precipitates out. Then filter, collect the filter cake, and dry it to obtain crystalline form C of solenoid estradiol.

4. The method for preparing crystal form C of soliestrol according to claim 3, characterized in that: Other solvents in step (1) include at least one of haloalkanes, alcohols, ketones, and esters.

5. The method for preparing crystal form C of soliestrol according to claim 4, characterized in that: The halogenated hydrocarbon solvents include at least one of dichloromethane, trichloromethane, and dichloroethane; the alcohol solvents include at least one of methanol, ethanol, and isopropanol; the ketone solvents include at least one of acetone, butanone, and cyclohexanone; and the ester solvents include at least one of ethyl acetate and methyl acetate.

6. The method for preparing crystal form C of soliestrol according to claim 3, characterized in that: In step (1), the mass-to-volume ratio of estradiol to dichloromethane or other solvents is 1:1 to 1:

15.

7. The method for preparing crystal form C of soliestrol according to claim 3, characterized in that: The dissolution temperature in step (1) is 50~80℃.

8. The method for preparing crystal form C of soliestrol according to claim 3, characterized in that: The stirring speed in step (2) is in the range of 200 rpm to 350 rpm.

9. The method for preparing crystal form C of soliestrol according to claim 3, characterized in that: The heat preservation temperature in step (3) is 10~45℃, and the stirring crystallization time is 1~8 hours.

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