Promestriene single crystal and preparation method thereof

Abstract

The invention provides a preparation method of a promestriene single crystal. The preparation method comprises the following steps: dissolving promestriene powder with isopropanol, filtering, transferring filtrate into a single crystal growth bottle, sealing the single crystal growth bottle with a plug, pricking the plug, and volatilizing under the conditions that the environment temperature is 30 + / -5 DEG C and the humidity is 50 + / -5% to obtain the promestriene single crystal. The invention provides a diffractable order single crystal for the first time, and the absolute configuration is directly analyzed through X-ray diffraction. The preparation method is simple to operate, only needs three steps of dissolving, filtering and volatilizing, and consumes 5-8 days. The prepared promestriene single crystal belongs to an orthorhombic crystal system and a P212121 space group, and the unit cell parameters of the promestriene single crystal are as follows: alpha is equal to 6.2174 (2), b is equal to 8.8913 (3), c is equal to 34.215 I (10), alpha is equal to 90 degrees, beta is equal to 90 degrees, gamma is equal to 90 degrees, and v is equal to 1891.44 (10).

Description

TECHNICAL FIELD

[0001] The present application relates to the field of medicine, in particular to a promestriene single crystal and a preparation method thereof. BACKGROUND

[0002] Promestriene is a local estrogen supplement, which is developed by Monaco Dalim S.p.A. (fully acquired by Merck & Co. in 1999), and approved by the United States in 1977. Its preparation mainly includes Colpotrophine marketed in France in 1975 and Delipoderm marketed in France in 1978. Promestriene, Colpotrophine (promestriene soft capsules and cream) and Delipoderm (chloroquine / promestriene vaginal tablets) have been collected in the Martindale Pharmaceutical Handbook. They have been clinically used in more than 40 countries for more than 40 years, and are still the first-line drugs for treating vaginal infections and atrophic vaginitis in many countries. Clinical practice shows that they are safe and effective, and well tolerated by patients.

[0003] The chemical structure is as follows: The synthesis process reported in the patent (GB1337198A) declared by Monaco Dalim S.p.A. in 1971 is as follows: In the first step, sodium metal is reacted with anhydrous ethanol to prepare sodium ethoxide to synthesize the intermediate 3-propoxy-17β-hydroxy-1,3,5(10)-estratriene. In the second step, sodium hydride and dimethyl sulfoxide are used to prepare dimethyl sulfoxide sodium solution, and then 3-propoxy-17β-hydroxy-1,3,5(10)-estratriene is treated with dimethyl sulfoxide sodium solution, and then methylated with dimethyl sulfate methyl ether to obtain promestriene. Finally, the promestriene product is obtained by repeated recrystallization with ethanol, and the molar yield is 78%.

[0004] Chinese patent CN110003298A discloses a synthesis method of promestriene. In the synthesis process, ethanol is used for dissolution, activated carbon is used for decolorization, cooling crystallization is used, filtration is used, and drying is used to obtain promestriene.

[0005] At present, a large number of literatures and patents have studied the synthesis process of promestriene. The absolute configuration of promestriene structure with five chiral carbon atoms is very important. Since some carbon atoms do not have hydrogen atoms, the relative configuration cannot be confirmed by two-dimensional nuclear magnetic resonance such as NOESY, so the only way to confirm the absolute configuration is to culture single crystals. The single crystal culture method and single crystal diffraction pattern of promestriene have not been found in the related literatures. Therefore, the prior art needs to be further developed. SUMMARY

[0006] In view of various deficiencies of the prior art, in order to solve the above problems, the present application provides a progestin single crystal and a preparation method thereof, and provides the following technical scheme: A preparation method of a progestin single crystal, comprising the following steps: dissolving progestin powder in isopropanol, filtering, transferring the filtrate to a single crystal growth bottle, sealing the single crystal growth bottle with a plug, punching a hole in the plug, and performing volatilization under the condition of an ambient temperature of 30±5 DEG C and a humidity of 50±5% to obtain the progestin single crystal.

[0007] Further, the ratio of isopropanol to progestin is 30-200 mL / g.

[0008] Further, the ratio of isopropanol to progestin is 50 mL / g.

[0009] Further, the progestin powder is dissolved in isopropanol and then subjected to ultrasonic treatment before filtering.

[0010] Further, a syringe filter head is used to filter into the single crystal growth bottle.

[0011] Further, the syringe filter head has a PTFE filter membrane with a pore size of 0.45 µm.

[0012] Further, the volatilization time is 5-8 days.

[0013] Further, the volatilization is performed under the condition of an ambient temperature of 30 DEG C and a humidity of 50%.

[0014] In addition, the present application also provides a progestin single crystal obtained by the above preparation method, wherein the progestin single crystal is in a rod-like form.

[0015] Further, the progestin single crystal belongs to an orthorhombic system and a P212121 space group, and has a unit cell parameter of: a = 6.2174(2) Å, b = 8.8913(3) Å, c = 34.2151(10) Å, α = 90°, β = 90°, γ = 90°, and v = 1891.44(10) Å.

[0016] Advantages: 1. The present application first provides a diffractive level single crystal, and the absolute configuration is directly analyzed by X-ray diffraction.

[0017] 2. The preparation method of the present application is simple to operate, and only three steps of dissolving, filtering and volatilizing are required, and the time consumption is 5-8 days. DETAILED DESCRIPTION

[0018] Figure 1 is a schematic diagram of a progestin single crystal system according to Embodiment 1 of the present application; Figure 2 is a schematic diagram of the asymmetric unit of the crystal structure of promegestone; Figure 3 is a schematic diagram of the unit cell of the crystal structure of promegestone. DETAILED DESCRIPTION

[0019] In order for those skilled in the art to better understand the technical solutions of the present application, the technical solutions of the present application will be described clearly and completely below in combination with the embodiments of the present application. Based on the embodiments in the present application, other similar embodiments obtained by those skilled in the art without making creative efforts should all belong to the scope of protection of the present application.

[0020] According to the embodiments of the present application, a preparation method of a promegestone single crystal is provided, which comprises the following steps: dissolving promegestone powder in isopropanol and then filtering, transferring the filtrate to a single crystal growth bottle, sealing the single crystal growth bottle with a plug and punching a hole on the plug, and then performing volatilization under the condition of an ambient temperature of 30±5°C and a humidity of 50±5% to obtain a promegestone single crystal. The present application first provides a diffractable single crystal, and the absolute configuration is directly analyzed by X-ray diffraction. The preparation method is simple to operate, only three steps of dissolving, filtering and volatilizing are needed, and the time consumption is 5-8 days. It is found in the present application that only isopropanol can be used to dissolve promegestone to obtain a diffractable promegestone single crystal. The prepared promegestone single crystal belongs to an orthorhombic system, a P212121 space group, and the unit cell parameters are as follows: a = 6.2174(2) Å, b = 8.8913(3) Å, c = 34.2151(10) Å, α = 90°, β = 90°, γ = 90°, and v = 1891.44(10) Å. The asymmetric unit of the promegestone single crystal structure is shown in Figure 2 , and the unit cell of the promegestone crystal structure is shown in Figure 3 .

[0021] Embodiment 1 100 mg of a compound promegestone sample was weighed into a 10 mL glass bottle, and 5 mL of isopropanol was added. After the solid sample was dissolved by ultrasonic oscillation (a frequency of 35 KHz and a power of 200 W), the sample was filtered by using a syringe filter head (a PTFE filter membrane with a pore size of 0.45 µm) and a 10 mL syringe, and the filtrate was transferred to a clean 10 mL single crystal growth bottle. The single crystal growth bottle was sealed with a PE single crystal bottle plug, a needle hole was punched on the plug, and then the single crystal growth bottle was placed in an environment with a temperature of 30°C and a humidity of 50% for slow volatilization. After slow volatilization for 7 days, rod-shaped single crystals and hairball-shaped crystals were observed to precipitate in the system, as shown in Figure 1A single crystal was selected from the rod-like crystal sample obtained from the above culture and subjected to single crystal X-ray diffraction characterization and crystal structure analysis. Single crystal diffraction data were collected at a temperature of 120 K using a Bruker D8 VENTURE single crystal diffractometer. Crystallographic data and refinement parameters are shown in Table 1.

[0022] Table 1 Crystallographic data and refinement parameters Example 2 A 100 mg sample of compound progestin was weighed into a 10 mL glass bottle, and 3 mL of isopropyl alcohol was added. After the solid sample was dissolved by ultrasonic oscillation (frequency of 35 KHz, power of 200 W), filtration was performed using a syringe filter head (PTFE filter membrane of 0.45 µm) and a 10 mL syringe, and the filtrate was transferred to a clean 10 mL single crystal growth bottle. The single crystal growth bottle was sealed with a PE single crystal bottle plug and a pinhole was punched in the plug, and then the single crystal growth bottle was placed in an environment with a temperature of 25 °C and a humidity of 45% for slow evaporation experiment. After 5 days of slow evaporation, rod-like single crystals and hairball-like crystals were observed to precipitate in the system.

[0023] Example 3 A 25 mg sample of compound progestin was weighed into a 10 mL glass bottle, and 5 mL of isopropyl alcohol was added. After the solid sample was dissolved by ultrasonic oscillation (frequency of 35 KHz, power of 200 W), filtration was performed using a syringe filter head (PTFE filter membrane of 0.45 µm) and a 10 mL syringe, and the filtrate was transferred to a clean 10 mL single crystal growth bottle. The single crystal growth bottle was sealed with a PE single crystal bottle plug and a pinhole was punched in the plug, and then the single crystal growth bottle was placed in an environment with a temperature of 35 °C and a humidity of 55% for slow evaporation experiment. After 8 days of slow evaporation, rod-like single crystals and hairball-like crystals were observed to precipitate in the system.

[0024] Comparative Example 1 Single crystal culture experiment using other solvents An appropriate amount of progestin sample was weighed into a 10 mL glass bottle, and a certain amount of corresponding solvent was added. After ultrasonic oscillation (frequency of 35 KHz, power of 200 W) to dissolve the sample, filtration was performed using a syringe filter head (PTFE filter membrane of 0.45 µm) and a 10 mL syringe, and the filtrate was transferred to a 10 mL bottle, which was sealed with a single crystal bottle plug and a small hole was punched in the plug. The environment was controlled at a temperature of 30 °C and a humidity of 50%, and the above system was placed for slow evaporation. According to the general experimental method, the following experiments were performed, and the specific experimental conditions and results are shown in Table 2.

[0025] Table 2 Results of single crystal culture experiment using other solvents The crystals obtained in Comparative Example 1 under various conditions were mostly needle-shaped or could not be obtained, and could not meet the size requirements of single crystal X-ray diffraction.

[0026] Comparative Example 2 Single crystal culture experiment by slow cooling method An appropriate amount of progestin sample was weighed into a 10 mL glass bottle, 0.5 mL of the corresponding solvent was added, and after ultrasonic treatment (frequency 35 KHz, power 200 W), the system was heated at 50°C for 30 minutes. If the sample was dissolved, an appropriate amount of sample was added to ensure that there was solid sample in the system. Then, while hot, filtration was performed using a syringe filter head (0.45 μm PTFE filter membrane) and a 10 mL syringe, and the hot filtrate was transferred to a 10 mL single crystal growth bottle. Finally, it was placed in an incubator at an initial temperature of 50°C for slow cooling. The specific experimental conditions and results are shown in Table 3.

[0027] Table 3 Results of single crystal culture experiment by slow cooling method The crystals obtained in Comparative Example 2 under 5 conditions were mostly needle-shaped or could not be obtained, and could not meet the size requirements of single crystal X-ray diffraction.

[0028] In addition, it should be understood that although the present specification is described in terms of embodiments, not every embodiment contains only one independent technical solution, and the description of the specification is only for the sake of clarity. Those skilled in the art should consider the specification as a whole, and the technical solutions in each embodiment can be appropriately combined to form other embodiments that those skilled in the art can understand.

Claims

1. A process for the preparation of monocrystalline crystals of promegestone, characterized in that, The method comprises the following steps: After dissolving the progestin powder in isopropanol, filtering, transferring the filtrate to a single crystal growth bottle, sealing the single crystal growth bottle with a plug, punching a hole on the plug, and volatilizing at ambient temperature of 30±5℃ and humidity of 50±5% to obtain the progestin single crystal.

2. The method for preparing prostaglandin single crystals according to claim 1, characterized in that, The ratio of isopropanol to progestin is 30-200 mL / g.

3. The method for preparing prostaglandin single crystals according to claim 2, characterized in that, The ratio of isopropanol to progestin is 50 mL / g.

4. The method for preparing prostaglandin single crystals according to claim 1, characterized in that, After dissolving the progestin powder in isopropanol, performing ultrasonic treatment, and then filtering.

5. The method for preparing prostaglandin single crystals according to claim 1, characterized in that, Filtering to a single crystal growth bottle by using a syringe filter head.

6. The method for preparing prostaglandin single crystals according to claim 5, characterized in that, The syringe filter head has a PTFE filter membrane with a pore size of 0.45 µm.

7. The method for preparing prostaglandin single crystals according to claim 1, characterized in that, The volatilization time is 5-8 days.

8. The method for preparing prostaglandin single crystals according to claim 1, characterized in that, The volatilization is performed at ambient temperature of 30℃ and humidity of 50%.

9. Progestin monocrystal crystals obtained by the process according to any one of claims 1 to 8, characterized in that, The progestin single crystal is in a rod-like shape.

10. The prosteglene monocrystalline crystal according to claim 9, characterized in that, The progestin single crystal belongs to an orthorhombic system and a P212121 space group, and has a unit cell parameter of α = 6.2174(2) Å, b = 8.8913(3) Å, c = 34.2151(10) Å, α = 90°, β = 90°, γ = 90°, and v = 1891.44(10) Å.

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