CN114644681A - Nemadeivir isopropanol solvate crystal form and preparation method thereof - Google Patents

Abstract

The application provides a crystal form of a nemadefovir isopropanol solvate and a preparation method thereof, relating to the technical field of medicines. The crystal form of the nemadefovir isopropanol solvate comprises peaks at 2 theta values of 4.54 +/-0.2 °, 6.55 +/-0.2 °, 7.51 +/-0.2 °, 8.18 +/-0.2 °, 8.88 +/-0.2 °, 10.82 +/-0.2 °, 12.63 +/-0.2 °, 12.85 +/-0.2 °, 14.41 +/-0.2 °, 15.46 +/-0.2 °, 16.52 +/-0.2 °, 17.11 +/-0.2 °, 18.25 +/-0.2 °, 18.97 +/-0.2 °, 19.57 +/-0.2 °, 19.83 +/-0.2 °, 20.50 +/-0.2 °, 21.31 +/-0.2 °, 22.28 +/-0.2 °, 22.57 +/-0.2 ° and 23.13 +/-0.2 ° under the condition of Cu-Kalpha radiation. The crystal form of the Nemadelter isopropanol solvate has higher purity and stability, and is suitable for development of new drugs and industrial production.

Description

Nemadeivir isopropanol solvate crystal form and preparation method thereof

Technical Field

The application relates to the technical field of medicines, in particular to a crystal form of a nemadefovir isopropanol solvate and a preparation method thereof.

Background

Nemadevir (Nirmatrelvir), chemical name: (1R,2S,5S) -N- [ (1S) -1-cyano-2- (2-oxo-pyrrolidin-3-yl) ethyl ] -3- [ (S) -3, 3-dimethyl-2- (trifluoroacetamido) butanoyl ] -6, 6-dimethyl-3-azabicyclo [3.1.0] hexane-2-amide having the formula:

。

nemadevir is a 3CL protease inhibitor which plays an important role in the life cycle of various coronaviruses, and has the potential advantage of acting on all new coronaviruses at present. The inhibitory activity IC50 of the nemadefovir single drug on the new coronavirus at the molecular level is 19nM, and in human airway epithelial cells, HeLa and A549 cells capable of expressing ACE2 protein, the inhibitory activity EC50 of the nemadefovir single drug on the virus is 62 nM, 99 nM and 56nM respectively, so that the nemadefovir single drug has good antiviral activity.

Nemadefovir dipivoxil has broad-spectrum anti-new corona virus capability, which means that the potential is very large, and although the virus still may mutate to escape from the action mechanism theoretically, the actual chance is very small. Currently, the clinical trial of pfeiffer is a combination therapy of nemadevir and ritonavir.

The literature (An Oral SARS-CoV-2 Mpro Inhibitor Clinical could for the Treatment of COVID-19) discloses a preparation and separation method of nemadefovir, in particular discloses a nemadefovir methyl tert-butyl ether solvate crystal and a nemadefovir crystal.

The form of the solid compound includes a crystalline form and an amorphous form. The crystal form of the compound also comprises a plurality of crystal form types such as single crystal of compound molecules and eutectic formed by the compound molecules and various solvents. Different crystal forms of the same compound have obvious differences in the aspects of solubility, melting point, density, stability and the like, so that the stability and the uniformity of the compound are influenced to different degrees. Designing and using the proper specific crystal form in the process can improve the performance of the compound, such as better operability and stability. The ability to purify a compound during purification is a significant benefit by using specific amorphous solids or crystalline forms of crystallization. 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

In order to solve the technical problems, the application provides a crystal form of a nemadefovir isopropanol solvate and a preparation method thereof.

One of the purposes of the application is to provide a crystal form of a nemadefovir isopropanol solvate, which has good stability.

The crystal form of the nemadefovir isopropanol solvate provided by the application comprises peaks at 2 theta values of 4.54 +/-0.2 °, 6.55 +/-0.2 °, 7.51 +/-0.2 °, 8.18 +/-0.2 °, 8.88 +/-0.2 °, 10.82 +/-0.2 °, 12.63 +/-0.2 °, 12.85 +/-0.2 °, 14.41 +/-0.2 °, 15.46 +/-0.2 °, 16.52 +/-0.2 °, 17.11 +/-0.2 °, 18.25 +/-0.2 °, 18.97 +/-0.2 °, 19.57 +/-0.2 °, 19.83 +/-0.2 °, 20.50 +/-0.2 °, 21.31 +/-0.2 °, 22.28 +/-0.2 °, 22.57 +/-0.2 ° and 23.13 +/-0.2 ° under the condition of Cu-Kalpha radiation.

One embodiment of the present application provides a crystalline form of the nemadefovir isopropanol solvate with an X-ray powder diffraction pattern expressed in degrees 2 θ using Cu-K α radiation as shown in fig. 1.

According to the crystal form of the isopropanol solvate of the nemadefovir, an endothermic peak appears at 123.7 ℃ in a differential scanning calorimetry analysis curve, and the peak of the endothermic peak appears at 128.9 ℃.

The thermogravimetric analysis curve of the crystal form of the nemadefovir isopropanol solvate provided by the application begins to lose weight when heated to 120.0 ℃.

The thermogravimetric analysis curve of the crystal form of the nemadefovir isopropanol solvate provided by the application is 9.73% of weight loss when the crystal form is heated to 190.0 ℃.

The thermogravimetric analysis curve of the crystal form of the nemadefovir isopropanol solvate provided by the application is 1.49% of weight loss when the crystal form is heated to 250.0 ℃.

The crystal form of the nemadefovir isopropanol solvate provided by the application starts to decompose at 300.0 ℃ and is completely decomposed at 350 ℃ according to a thermogravimetric analysis curve.

One embodiment of the present application provides a crystalline form of the isopropanol solvate of nemadefovir, wherein a Differential Scanning Calorimetry (DSC) curve and a thermogravimetric analysis curve are shown in FIG. 2.

The melting point of the crystal form of the nimatvir isopropanol solvate provided by the application is 121.5-127.7 ℃ measured by a capillary method.

The second purpose of the application is to provide a preparation method of the crystal form of the nemadefovir isopropanol solvate, the preparation method has simple process route and low cost, and the prepared nemadefovir isopropanol solvate has higher yield and purity and is suitable for new drug development and industrial production.

The preparation method of the crystal form of the isopropanol solvate of the nemadefovir comprises the steps of dissolving a crude product of the nemadefovir with isopropanol, crystallizing at 0-5 ℃, filtering, and drying under reduced pressure to obtain the isopropanol solvate of the nemadefovir. Specifically, the dissolving of the crude nemadevir product in isopropanol means that: adding the crude product of the nemadevir into isopropanol, heating to 70-80 ℃, and stirring; wherein the volume ratio of the nemadefovir crude product to the isopropanol is 1 (5-10); the crystallization time is 8-16 h.

In the present application, the volume ratio of the crude nemadefovir product to isopropanol refers to the ratio between the mass of the crude nemadefovir product (in g) and the volume of isopropanol (in mL).

The crude nemadevir product used in the preparation method of the application is nemadevir methyl tert-butyl ether solvate.

In the present application, the purity of the crystalline form refers to the content of the crystalline form after removal of other crystalline or amorphous forms of the nemadefovir isopropanol solvate and other impurities, as measured by HPLC.

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 values provided cannot be taken as absolute values.

In summary, the present application has the following beneficial technical effects:

1. the crystal form of the isopropanol solvate of the nemadefovir provided by the application has higher purity and stability, and is suitable for development of new drugs and industrial production.

2. According to the preparation method of the crystal form of the isopropanol solvate of the nemadefovir, the yield of the prepared isopropanol solvate of the nemadefovir can reach more than 90%, the purity can reach more than 99.8%, and the single impurity content is less than 0.1%.

3. The preparation method of the crystal form of the isopropanol solvate of the nemadefovir provided by the application has the advantages of simple process route and low cost, and is suitable for industrial production.

Drawings

Figure 1 is an XRPD pattern of the crystalline form of the nemadefovir isopropanol solvate prepared in example 1;

figure 2 is a DSC diagram and TGA diagram of the crystalline form of the nemadefovir isopropanol solvate prepared in example 1;

figure 3 is a High Performance Liquid Chromatography (HPLC) chart of the nemadevir isopropanol solvate crystalline form prepared in example 1;

figure 4 is a hydrogen spectrum (H-NMR) plot of the crystalline form of the nemadefovir isopropanol solvate prepared in example 1.

Detailed Description

Embodiments of the present invention will be described in detail below with reference to examples, but those skilled in the art will appreciate that the following examples are only illustrative of the present invention and should not be construed as limiting the scope of the present invention. The 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.

In the present application:

XRPD: x ray powder diffraction (also called XRD);

DSC: differential scanning calorimetry;

TGA: thermovirometric Analysis, Thermogravimetric Analysis;

the XRPD test uses an X-ray diffractometer of the Empyrean type from Panalytical corporation. Appropriate amount of samples were spread evenly on a single crystal silicon sample pan with the test parameters as shown in table 1.

TABLE 1XRPD test parameters

DSC and TGA spectra are respectively collected on a TA Discovery DSC 2500 differential scanning calorimeter and a TA Discovery TGA 5500 thermogravimetric analyzer, and the test parameters are shown in Table 2.

TABLE 2DSC and TGA test parameters

Example 1

Adding 10Kg of crude nemadevir into 90L of isopropanol, stirring and heating to 70 ℃ for complete dissolution, stopping heating, continuing stirring, cooling and crystallizing, cooling to 1 ℃, keeping the temperature for crystallizing for 16h, filtering, and drying under reduced pressure at 70 ℃ for 16h to obtain 9.1Kg of refined product, wherein the yield is as follows: 96%, purity (HPLC assay, as shown in fig. 3): 99.96 percent and less than 0.1 percent of single impurity. The refined product was subjected to XRPD, and the obtained XPRD pattern is shown in FIG. 1, and the peak information thereof is shown in Table 3. Typical DSC and TGA are shown in FIG. 2, and have melting points of 121.5 ℃ to 127.7 ℃ (the melting range is 123.7 ℃ to 140 ℃ as determined by DSC). The hydrogen spectrum is shown in figure 4, and the obtained refined product is the crystal form of the isopropanol solvate of the nemadevir. Specific information of the hydrogen spectrogram is as follows:

¹H NMR (400 MHz, DMSO)δ9.41 (d, J = 8.4 Hz, 1H), 9.02 (d, J = 8.5 Hz, 1H), 7.67 (s, 1H), 4.97 (ddd, J = 10.9, 8.6, 5.2 Hz, 1H), 4.42 (d, J = 8.5 Hz, 1H), 4.34 (d, J = 4.2 Hz, 1H), 3.91 (dd, J = 10.3, 5.5 Hz, 1H), 3.77 (dtd, J = 12.2, 6.1, 4.2 Hz, 1H), 3.70 (d, J = 10.4 Hz, 1H), 3.14 (t, J = 9.0 Hz, 1H), 3.04 (dt, J = 16.5, 8.3 Hz, 1H), 2.44 -2.34 (m, 1H), 2.21 -2.00 (m, 2H), 1.78 -1.65 (m, 2H), 1.57 (dd, J = 7.5, 5.5 Hz, 1H), 1.32 (d, J = 7.6 Hz, 1H), 1.05 (s, 3H), 1.03 (s, 6H), 0.98 (s, 9H), 0.85 (s, 3H)

TABLE 3 powder diffraction data Table for sample obtained in example 1

Example 2

Adding 1Kg of crude nemadevir product into 7L of isopropanol, stirring and heating to 75 ℃ for complete dissolution, stopping heating, continuing stirring, cooling and crystallizing, cooling to 2 ℃, keeping the temperature for crystallizing for 12h, filtering, and drying under reduced pressure at 70 ℃ for 12h to obtain 933g of refined product, wherein the yield is as follows: 98%, purity: 99.81 percent and less than 0.1 percent of single impurity. The refined product obtained was subjected to XRPD testing, and XPRD was substantially in accordance with the results of FIG. 1. The refined product is the crystal form of the isopropanol solvate of the nemadefovir dipivoxil.

Example 3

Adding 100 g of crude nemadevir into 500 mL of isopropanol, stirring and heating to 80 ℃ for complete dissolution, stopping heating, continuously stirring, cooling and crystallizing, cooling to 0 ℃, keeping the temperature for crystallizing for 8 hours, filtering, and drying under reduced pressure at 70 ℃ for 10 hours to obtain 90.5 g of refined product, wherein the yield is as follows: 95%, purity: 99.91 percent and less than 0.1 percent of single impurity. The refined product was subjected to XRPD testing, and XPRD was substantially in accordance with the results of fig. 1. The refined product is the crystal form of the isopropanol solvate of the nemadefovir dipivoxil.

Example 4

Stability test

Stability tests were performed on the crystalline forms of the nemadefovir isopropanol solvate prepared in examples 1-3, with the test results shown in 4:

TABLE 4 stability test results

From the stability test results in table 4, it can be seen that the stability of the crystal form of the nemadefovir isopropanol solvate of the present application is mainly examined and compared with the initial time (0 month), the comparison results are all no obvious change, the purity is greater than 99%, the single impurity is less than 0.1%, and the stability is relatively stable.

In table 4:

"Total hetero" means: the total amount and the mass percentage of impurities contained in the nemadevir isopropanol solvate crystal form are as follows;

"moisture" means: the water content and mass percentage of the crystal form of the Nemadeivir isopropanol solvate are shown in the specification;

"content" means: the crystal form of the Nemadelter isopropanol solvate contains the amount (by taking a standard product as a reference) and the mass fraction of the effective components of the Nemadelter isopropanol solvate;

melting point determination method: adding the nerametevir isopropyl alcohol solvate 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 China pharmacopoeia 2020 edition;

the water content measuring method comprises the following steps: an appropriate amount of the isopropanol solvate of the nemadefovir prepared by the application is determined according to a moisture determination method (0832 first method 1 of the four general rules of the Chinese pharmacopoeia 2020 edition);

the method for measuring the content of single impurity and total impurity comprises the following steps: measuring by high performance liquid chromatography (China pharmacopoeia 2020 edition four-part general regulation 0512).

The above embodiments are preferred embodiments of the present application, and the protection scope of the present application is not limited by the above embodiments, so: all equivalent changes made according to the structure, shape and principle of the present application shall be covered by the protection scope of the present application.

Claims (10)

1. A crystalline form of nemadefovir isopropanol solvate characterized by an X-ray powder diffraction pattern comprising peaks at 2 Θ values of 4.54 ± 0.2 °, 6.55 ± 0.2 °, 7.51 ± 0.2 °, 8.18 ± 0.2 °, 8.88 ± 0.2 °, 10.82 ± 0.2 °, 12.63 ± 0.2 °, 12.85 ± 0.2 °, 14.41 ± 0.2 °, 15.46 ± 0.2 °, 16.52 ± 0.2 °, 17.11 ± 0.2 °, 18.25 ± 0.2 °, 18.97 ± 0.2 °, 19.57 ± 0.2 °, 19.83 ± 0.2 °, 20.50 ± 0.2 °, 21.31 ± 0.2 °, 22.28 ± 0.2 °, 22.57 ± 0.2 °, 23.13 ± 0.2 ° in the presence of Cu-ka radiation.

2. The crystalline form of nemadefovir isopropanol solvate according to claim 1, characterized in that it has an X-ray powder diffraction pattern as shown in figure 1.

3. The crystalline form of nemadefovir isopropanol solvate according to claim 1, characterized in that it has an endothermic peak at 123.7 ℃ and a peak at 128.9 ℃ in its differential scanning calorimetry curve.

4. The crystalline form of nemadefovir isopropanol solvate according to claim 1, characterized in that its thermogravimetric analysis curve starts to lose weight when heated to 120.0 ℃.

5. The crystalline form of nemadefovir isopropanol solvate according to claim 1, characterized in that its thermogravimetric analysis curve loses 9.73% of its weight when heated to 190.0 ℃;

its thermogravimetric analysis curve shows that when it is heated to 250.0 deg.C, it has 1.49% of weight loss.

6. The crystalline form of nemadefovir isopropanol solvate according to claim 1, characterized in that its thermogravimetric analysis curve starts to decompose at 300.0 ℃ and completely at 350 ℃.

7. The crystalline form of nemadefovir isopropanol solvate according to any of claims 3-6, characterized in that its differential scanning calorimetry curve and its thermogravimetric analysis curve are shown in figure 2.

8. The crystalline form of nemadefovir isopropanol solvate according to claim 1, characterized in that the melting point as measured by capillary method is in the range of 121.5-127.7 ℃.

9. A method for preparing a crystalline form of the nelamavir isopropanol solvate according to any one of claims 1 to 8, characterized in that a crude product of the nelamavir is dissolved in isopropanol, crystallized at 0 to 5 ℃, filtered, and dried under reduced pressure to obtain the nelamavir isopropanol solvate.

10. The method according to claim 9, wherein the dissolving of the crude nemadevir product in isopropanol means: adding the crude product of the nemadevir into isopropanol, heating to 70-80 ℃, and stirring;

wherein the volume ratio of the nemadefovir crude product to the isopropanol is 1 (5-10);

the crystallization time is 8-16 h.

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