CN115433132A

CN115433132A - Enconazole crystal form, preparation method and application

Info

Publication number: CN115433132A
Application number: CN202211286225.9A
Authority: CN
Inventors: 彭康洲; 郑春红; 刘祥; 张锐; 张鹤营; 黄金斌; 向光强
Original assignee: WUHAN HVSEN BIOTECHNOLOGY CO Ltd
Current assignee: Hubei Hesheng Biopharmaceutical Co.,Ltd.
Priority date: 2022-10-20
Filing date: 2022-10-20
Publication date: 2022-12-06

Abstract

The invention discloses an enilconazole crystal form, a preparation method and application, wherein the crystal form has characteristic peaks at 9.3045, 13.3046, 14.8313, 20.2166, 20.6254, 20.7572, 21.0752, 22.7064, 22.9297, 23.4409, 23.6543, 24.4229, 25.2438, 27.7342, 28.4334 and 28.5207 in an X-ray powder diffraction pattern expressed by Cu-K alpha radiation and 2 theta +/-0.2-degree diffraction angle. The enilconazole crystal form has high purity and stability, is suitable for development of new drugs and industrial production, and has high application value.

Description

Enconazole crystal form, preparation method and application

Technical Field

The invention relates to the technical field of enilconazole preparation, in particular to an enilconazole crystal form, a preparation method and application.

Background

Enilconazole (Enilconazole), also known as imazalil (imazalil), is an imidazole fungicide developed in 1973 by the company popsonian, belgium (Janssen), and formally approved for use in the united states in 1979. Enconazole is used as a broad-spectrum antifungal drug, is mainly used for treating dermatophyte infection of cows, horses and dogs in veterinary drugs, and is sold in various countries or regions all over the world at present. The chemical name of the compound is 1- [ (2 RS) -2- (2,4-dichlorophenyl) -2- (2-propenyloxy) ethyl ] -1H-imidazole, and the structural formula is as follows:

most of the enoconazole is used as an agricultural bactericide in a liquid form, and the existing synthetic enoconazole is yellow to brown oily liquid, contains a large amount of impurities which are difficult to remove in the synthesis process, is unstable during storage, and influences the prospect of the enoconazole serving as a bactericidal medicine for livestock.

The research and optimization of the solid form are known to better improve the performance of the compound, the improvement of the purity is of great significance for veterinary drugs, and at present, no literature report on the crystal form research of enilconazole exists temporarily.

The morphology of a solid compound includes a crystal form and an amorphous form, and different crystal forms of the same compound have significant differences in solubility, melting point, density, stability and the like, thereby affecting the stability and homogeneity of the compound to different degrees. The involvement and use of the appropriate specific crystalline form in the process can improve the properties of the compound, such as better handling and stability. Therefore, comprehensive systematic crystal form and amorphous screening are carried out in the research and development of the pharmaceutical technology, and the selection of the most suitable developed solid form is one of important research contents which cannot be ignored.

Disclosure of Invention

The invention aims to provide an enilconazole crystal form and a preparation method thereof, overcomes the defects of low purity, poor stability and the like of enilconazole liquid in the prior art, and promotes the development and industrial production process of enilconazole as a medicament.

In view of the above, the technical scheme of the invention is as follows:

an enilconazole crystalline form having characteristic peaks in an X-ray powder diffraction pattern expressed as Cu-K α radiation at 2 θ ± 0.2 ° diffraction angles at 9.3045, 13.3046, 14.8313, 20.2166, 20.6254, 20.7572, 21.0752, 22.7064, 22.9297, 23.4409, 23.6543, 24.4229, 25.2438, 27.7342, 28.4334, 28.5207.

Further, the crystalline form of eniconazole has an X-ray powder diffraction pattern as shown in fig. 1.

Further, a differential scanning calorimetry analysis curve of the enilconazole crystal form shows a first endothermic peak at 50-53 ℃, and the peak value of the first endothermic peak shows 53-55 ℃.

Further, the thermogravimetric analysis curve of the crystalline form of eniconazole starts to decompose at 150 ℃ and completely decomposes at 210 ℃.

The invention also aims to provide a method for preparing the enilconazole crystal form, which comprises the steps of adding a crude enilconazole product into a solvent, heating and stirring for the first time, and filtering; and (3) heating and stirring the filtered product for the second time, cooling after dissolving, stirring and crystallizing, filtering, and drying under reduced pressure to obtain the product.

Further, the solvent is one of dichloromethane, butyl acetate, methyl tert-butyl ether, n-hexane, ethyl acetate or toluene.

Further, the volume ratio of the crude enilconazole product to the solvent is 1: (0.5-10).

Further, the crystallization process time is 2-8h.

The invention also provides application of the enilconazole crystal form in preparation of bactericidal or bacteriostatic medicines.

Still another object of the present invention is to provide a method for detecting the crystalline form of enilconazole, wherein the suspected crystal is subjected to X-ray powder diffraction detection, and the obtained X-ray powder diffraction pattern is compared with the X-ray powder diffraction pattern shown in fig. 1.

The beneficial effects of the present invention include but are not limited to:

1. the eniconazole crystal form provided by the invention has higher purity and stability, is suitable for new drug development and industrial production, and has higher application value.

2. According to the preparation method of the enilconazole crystal form, the yield of the prepared enilconazole can reach more than 90%, the purity of the enilconazole crystal can reach more than 99.9%, and the single impurity content of the enilconazole crystal form is less than 0.1% through multiple times of thermal stirring and filtering. And the preparation method has simple process route and low cost and is suitable for industrial production.

3. The enilconazole crystal form provided by the invention can be used for detecting enilconazole suspected crystals, and a conclusion can be obtained according to the comparison of characteristic peaks in X-ray powder, so that the method is convenient and rapid.

Drawings

Fig. 1 is an XRPD pattern of the crystalline form of eniconazole prepared in example 1 of the present invention.

Fig. 2 is a DSC diagram of the crystalline form of eniconazole prepared in example 1 of the present invention.

Fig. 3 is a TGA diagram of an eniconazole crystalline form prepared in example 1 of the present invention.

FIG. 4 is a H-NMR chart of an eniconazole crystal form prepared in example 1 of the present invention.

FIG. 5 is an XRPD pattern versus crystalline forms of eniconazole prepared in examples 1-6 of the present invention.

FIG. 6 is a DSC of crystalline forms of eniconazole prepared in examples 1-6 of the present invention in comparison.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

In one embodiment of the invention, the crystalline form of eniconazole has characteristic peaks at 9.3045, 13.3046, 14.8313, 20.2166, 20.6254, 20.7572, 21.0752, 22.7064, 22.9297, 23.4409, 23.6543, 24.4229, 25.2438, 27.7342, 28.4334, 28.5207 in an X-ray powder diffraction pattern expressed by Cu-K alpha radiation and 2 theta +/-0.2 DEG diffraction angle, has high purity and stability, is suitable for new drug development and industrial production, and has high application value.

In a preferred embodiment, the crystalline form of eniconazole has an X-ray powder diffraction pattern as shown in figure 1.

In one embodiment, the differential scanning calorimetry curve of the crystalline form of eniconazole exhibits a first endotherm at 50-53 ℃ and the peak of the first endotherm occurs at 53-55 ℃; preferably, the sample has a differential scanning calorimetry curve and a characteristic peak as shown in FIG. 2.

In one embodiment, the thermogravimetric analysis curve of the crystalline form of eniconazole begins to decompose at 150 ℃ and is completely decomposed at 210 ℃; preferably with a thermogravimetric analysis curve as shown in figure 3.

In another embodiment of the present invention, a method for preparing the crystalline form of enilconazole described above is provided, wherein a crude enilconazole is added to a solvent, heated and stirred for the first time, and filtered; and (3) heating and stirring the filtered product for the second time, cooling after dissolving, stirring and crystallizing, filtering, and drying under reduced pressure to obtain the product.

Based on the knowledge of those skilled in the art, the solvents used for crystallization in the prior art are various, and the mixed solvents composed of solvents with different types and proportions cannot be counted, and the design of crystallization is based on experience, so that the obtained crystal form cannot be predicted according to crystallization conditions.

In one embodiment, in the preparation method of the enilconazole crystal form, the solvent is one of dichloromethane, butyl acetate, methyl tert-butyl ether, n-hexane, ethyl acetate or toluene. In a preferred embodiment, the volume ratio of the crude enilconazole to the solvent is 1: (0.5-10). And the solvent lost due to the first filtration is supplemented during the secondary heating and stirring, so that the impurity precipitation capacity is ensured. In the process, the stirring time, the heating temperature and the heating time are determined by fully dissolving the crude product in the solvent and fully separating out impurities, the crystallization process is carried out in a cooling process, the crystallization time influences the yield of the crystal form, and the preferable time is 2-8h. The yield of the prepared enoconazole crystal form can reach more than 90%, the purity reaches more than 99.9%, and the single impurity is less than 0.1%.

In one embodiment, a bactericidal or bacteriostatic drug is provided, which comprises the crystal of the above embodiment and a pharmaceutically acceptable excipient, preferably consists of the crystalline form of enilconazole of the above embodiment and a pharmaceutically acceptable excipient. Pharmaceutically acceptable excipients include, but are not limited to, common fillers, stabilizers, diluents, adjuvants or other formulation excipients, based on routine selection in the art.

In one embodiment, a method for detecting the crystalline form of enilconazole is provided, wherein an X-ray powder diffraction detection is performed on a suspected crystal, and the obtained X-ray powder diffraction pattern is compared with an X-ray powder diffraction pattern shown in figure 1. Those skilled in the art can obtain the diffraction pattern of the crystal using X-ray powder diffraction detection equipment, which is commercially available. The detection conditions may also be well known to those skilled in the art, such as voltage: current, cu-ka radiation, etc. According to parameters such as line position, line height, relative abundance and/or interplanar distance d (usually expressed) of the spectrum, a person skilled in the art can compare whether the suspected crystal is the enilconazole crystal form provided by the invention.

The following examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products commercially available.

XRPD: x-ray powder diffraction (also known as XRPD);

DSC: differential Scanning Calorimetry, differential Scanning Calorimetry;

TGA: thermal gravimetric analysis, thermogravimetric analysis;

XRPD testing used an EMPYREAN X-ray diffractometer from PANalytical b.v. netherlands. Appropriate amount of samples were spread evenly on the single crystal silicon sample plate and the test parameters are shown in table 1.

Table 1: XRPD test conditions

DSC and TGA spectra were collected on a Diamond DSC differential scanning calorimeter of PerkinElmer, USA, and a Pyris1TGA thermogravimetric analyzer of PerkinElmer, USA, and the test parameters are shown in Table 2.

Table 2: DSC and TGA test parameters

	DSC	TGA
			Sample plate	Hollow AI crucible (tablet)	Air AI crucible (tablet)
Temperature range (. Degree.C.)	30-60	30-500
			Rate of temperature rise (. Degree. C./min)	5	5
Protective gas	High purity N ₂	High purity N ₂

Crude enilconazole as used herein refers to the compound as an oil, without crystallization. In the application, because the eniconazole cannot be completely dissolved in the solvent, heating and stirring are required to remove impurities contained in the eniconazole crude product, so that the eniconazole is crystallized again, and the eniconazole crystal form provided by the application is prepared through subsequent steps.

In the present application, the purity of the crystalline form refers to the content of the crystalline form after removing crystalline or amorphous enoconazole and other impurities, and the determination method is determined by GC.

It will be appreciated that slightly different melting point readings may be given with different types of equipment or with different test conditions. The exact values of the melting points of the different crystal forms will be influenced by the purity of the compound, the weight of the sample, the heating rate, the particle size and the calibration and maintenance of the test equipment. The values provided cannot be taken as absolute values.

It should be understood that slightly different XRPD patterns and peaks may be given with different types of equipment or with different test conditions. The spectra, peak values and relative intensities of the various diffraction peaks of the different crystal forms will be affected by the purity of the compound, the pre-treatment of the sample, the scanning speed, the particle size and the calibration and maintenance of the test equipment. The numerical values provided cannot be taken as absolute values.

Example 1

Adding 20g of the oily crude product of enilconazole into 10mL of ethyl acetate, stirring and heating to 38 ℃, preserving heat and stirring for 1.0h, performing heat filtration, supplementing ethyl acetate lost by suction filtration to 10mL, heating and stirring to 36 ℃, preserving heat and stirring for 1.0h, cooling to 5 ℃, preserving heat and stirring for 4h, performing crystallization, filtering, and drying at 30 ℃ under reduced pressure for 24h to obtain 18g of refined product, wherein the yield is as follows: 90%, purity (GC assay): 99.95 percent and less than 0.1 percent of single impurity. The obtained refined product was subjected to XRPD, and the obtained XRPD pattern is shown in FIG. 1, and the peak information thereof is shown in Table 3. A typical DSC diagram is shown in fig. 2, with a melting point of 53 ℃ and a melting range of 52..0-54.0 ℃ as interpreted from fig. 2. A typical TGA profile is shown in figure 3. The resulting enilconazole crystals were free of water or other solvents as evidenced by the H-NMR plot of fig. 4. The obtained refined product is the enilconazole crystal form.

Table 3: powder diffraction data of samples obtained in example 1

Example 2

Adding 100g of oily enoconazole crude product into 200mL of dichloromethane, stirring and heating to 32 ℃, preserving heat and stirring for 0.5h, performing heat filtration, supplementing dichloromethane lost by suction filtration to 200mL, heating and stirring to 30 ℃, preserving heat and stirring for 0.5h, cooling to 8 ℃, preserving heat and stirring for 8h, crystallizing, filtering, and drying at 30 ℃ under reduced pressure for 12h to obtain 85g of refined product, yield: 85%, purity (GC assay): 99.92 percent and less than 0.1 percent of single impurity. The obtained refined product was subjected to XRPD, XPRD as shown in FIG. 5 (EKZ-1), and information on the peak value of the diffraction pattern as shown in Table 4, which was substantially in accordance with the results of FIG. 1. The obtained refined product is the enilconazole crystal form.

Table 4: example 2 powder diffraction data of samples

Example 3

Adding 500g of oily enoconazole crude product into 1500mL of butyl acetate, stirring and heating to 38 ℃, preserving heat and stirring for 1.0h, carrying out heat filtration, supplementing butyl acetate lost by suction filtration to 1500mL, heating and stirring to 40 ℃, preserving heat and stirring for 0.5h, cooling to 0 ℃, preserving heat and stirring for 5h, crystallizing, filtering, and drying at 30 ℃ under reduced pressure for 30h to obtain 1380g of refined product, yield: 92%, purity (GC assay): 99.91 percent and less than 0.1 percent of single impurity. The refined product thus obtained was subjected to XRPD (EKZ-2, FIG. 5), and the peak information of XPRD pattern was substantially in accordance with the results shown in FIG. 1, as shown in Table 5. The obtained refined product is the enilconazole crystal form.

Table 5: powder diffraction data of sample obtained in example 3

Example 4

Adding 200g of oily crude enilconazole into 2000mL of methyl tert-butyl ether, stirring and heating to 52 ℃, preserving heat and stirring for 1.0h, performing heat filtration, supplementing the lost methyl tert-butyl ether to 2000mL, heating and stirring to 55 ℃, preserving heat and stirring for 1.0h, cooling to 22 ℃, preserving heat and stirring for 5h, performing crystallization, filtering, and drying under reduced pressure at 30 ℃ for 10h to obtain 188g of refined product, wherein the yield is as follows: 94%, purity (GC assay): 99.93 percent and less than 0.1 percent of single impurity. The obtained refined product was subjected to XRPD (EKZ-3, FIG. 5), and the information on the peak value of the diffraction pattern is shown in Table 6, and substantially agrees with the result of FIG. 1. The obtained refined product is the enilconazole crystal form of the application.

Table 6: powder diffraction data of sample obtained in example 4

Example 5

Adding 30g of the oily crude product of enilconazole into 600mL of n-hexane, stirring and heating to 67 ℃, keeping the temperature and stirring for 1.0h, performing heat filtration, supplementing the n-hexane lost by suction filtration to 600mL, heating and stirring to 69 ℃, keeping the temperature and stirring for 1.0h, cooling to 30 ℃, keeping the temperature and stirring for 2h for crystallization, filtering, and drying at 30 ℃ under reduced pressure for 10h to obtain 28.5g of refined product, wherein the yield is as follows: 95%, purity (GC assay): 99.90 percent and less than 0.1 percent of single impurity. The resulting refined product was subjected to XRPD (EKZ-4, FIG. 5), and the information on the peak value of the diffraction pattern is shown in Table 7, which is substantially in accordance with the results shown in FIG. 1. The obtained refined product is the enilconazole crystal form.

Table 7: powder diffraction data sheet of sample obtained in example 5

Example 6

Adding 1000g of oily enoconazole crude product into 2000mL of toluene, stirring and heating to 55 ℃, preserving heat and stirring for 0.5h, performing heat filtration, supplementing toluene lost by suction filtration to 2000mL, heating and stirring to 60 ℃, preserving heat and stirring for 0.5h, cooling to 30 ℃, preserving heat and stirring for 5h, performing crystallization, filtering, and drying at 30 ℃ under reduced pressure for 36h to obtain 890g of refined product, wherein the yield is as follows: 89%, purity (GC assay): 99.95 percent and less than 0.1 percent of single impurity. The obtained refined product was subjected to XRPD (EKZ-6, FIG. 5), and the information on the peak value of the diffraction pattern is shown in Table 8, and substantially agrees with the result of FIG. 1. The obtained refined product is the enilconazole crystal form of the application.

Table 8: example 6 powder diffraction data sheet of the sample

In the above examples 1 to 6, the crystalline forms of eniconazole prepared by different solvents were numbered as shown in table 9, and XRPD diffraction pattern peak information and DSC curve characteristics of the crystalline forms were compared as shown in fig. 5 and 6, respectively. The high consistency of the diffraction pattern peak value information can be easily seen; in a DSC curve, the first endothermic peak appears in the enilconazole crystal form at 50-53 ℃, the peak value of the first endothermic peak appears at 53-55 ℃, and the enilconazole crystal form has consistency.

Table 9: sample information

Crystallization solvent	Sample numbering	Corresponding embodiment
			Methylene dichloride	EKZ-1	Example 2
Acetic acid butyl ester	EKZ-2	Example 3
			Methyl tert-butyl ether	EKZ-3	Example 4
N-hexane	EKZ-4	Example 5
			Ethyl acetate	EKZ-5	Example 1
Toluene	EKZ-6	Example 6

Stability test of Crystal form

Stability tests were performed on the crystalline forms of eniconazole prepared in examples 1 to 6, and the relevant indexes were measured by leaving for different times, and the test results are shown in table 10.

Table 10: results of stability test

Compared with the initial (0 month), the stability test result of the Enconazole crystal form of the application shows that the indexes of the key investigation items of the Enconazole crystal form are not obviously changed compared with the initial test time (0 month), the purity is more than 99.0%, the single impurity content is less than 0.1%, and the stability is relatively stable.

In table 9:

"Total hetero" means: the total amount and the mass percentage of impurities contained in the enilconazole crystal form;

"moisture" means: the amount and the mass percentage of the water contained in the eniconazole crystal form;

"content" means: the content (taking a standard as a reference) and the mass fraction of the effective components of the enilconazole in the enilconazole crystal form;

melting point determination method: adding the eniconazole prepared by the method into a capillary tube to form a compact column with the height of about 3mm, and testing the melting point according to the four-part general rule 0612 of the 2020 edition of Chinese pharmacopoeia;

the water content measuring method comprises the following steps: taking a proper amount of eniconazole prepared by the application, and measuring according to a moisture determination method (0823 first method 1 of the four ministry of general rules of China pharmacopoeia 2020); the single impurity and total impurity content determination method comprises the following steps: measured according to gas chromatography (China pharmacopoeia 2020 edition four-part general rules 0512).

Finally, although the present invention has been described in detail by way of general description and specific examples, the above examples are only intended to illustrate the technical solutions of the present invention, but not to limit the same; while the invention has been described in detail and with reference to the foregoing embodiments, it will be understood by those skilled in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some or all of the technical features may be equivalently replaced; and the modifications or the substitutions do not make the essence of the corresponding technical solutions depart from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. An enilconazole crystalline form characterized by characteristic peaks in an X-ray powder diffraction pattern expressed as Cu-K α radiation at 2 θ ± 0.2 ° diffraction angles at 9.3045, 13.3046, 14.8313, 20.2166, 20.6254, 20.7572, 21.0752, 22.7064, 22.9297, 23.4409, 23.6543, 24.4229, 25.2438, 27.7342, 28.4334, 28.5207.

2. The crystalline form of eniconazole according to claim 1, characterized by the X-ray powder diffraction pattern as shown in figure 1.

3. The crystalline form of eniconazole according to claim 1, wherein a differential scanning calorimetry curve shows a first endothermic peak at 50-53 ℃ and the peak of the first endothermic peak at 53-55 ℃.

4. The crystalline form of eniconazole according to claim 1, wherein the thermogravimetric analysis curve begins to decompose at 150 ℃ and is completely decomposed at 210 ℃.

5. A method for preparing the crystalline form of enilconazole as defined in any one of claims 1 to 4, wherein a crude enilconazole is added to a solvent, stirred under heating for a first time and filtered; and (3) heating and stirring the filtered product for the second time, cooling after dissolving, stirring and crystallizing, filtering, and drying under reduced pressure to obtain the product.

6. The method of claim 5, wherein the solvent is one of dichloromethane, butyl acetate, methyl tert-butyl ether, n-hexane, ethyl acetate, or toluene.

7. The preparation method of claim 5, wherein the volume ratio of the crude enilconazole to the solvent is 1: (0.5-10).

8. The method according to claim 5, wherein the time of the crystallization process is 2 to 8 hours.

9. Use of the crystalline form of enilconazole of any one of claims 1 to 4 in the preparation of a bactericidal or bacteriostatic medicament.

10. A method for detecting the crystalline form of enilconazole as defined in claim 1 or 2, wherein the suspected crystals are subjected to an X-ray powder diffraction detection and the X-ray powder diffraction pattern obtained is compared with an X-ray powder diffraction pattern shown in fig. 1.