Billidofovir compound and preparation method thereof

Abstract

The invention provides a brilincidofovir compound and a preparation method thereof, and the method comprises the following steps: sequentially adding a 0.05-0.2 mol / L sodium hydroxide solution and brilincidofovir into a reaction pot, starting stirring, heating to 40-44 DEG C, stirring and dissolving; cooling to 20-24 DEG C, slowly adding a mixed solution of ethanol and trichloromethane while stirring, and continuously stirring after adding to obtain a material; and transferring to a crystallization kettle, cooling to 3-5 DEG C at a speed of 3-3.6 DEG C / min, filtering to obtain a filter cake, drying, and crushing the filter cake to obtain the brilincidofovir trihydrate. The inventor accidentally obtains the novel brilincidofovir compound and the preparation method thereof in a large number of research processes on brilincidofovir for a long time, and the brilincidofovir compound prepared by the method has extremely high quality stability, the hygroscopicity is reduced, and the solubility is improved.

Description

Technical Field

[0001] The present invention relates to the field of compounds, and in particular to a brincidofovir compound and a preparation method thereof. Background Art

[0002] Brincidofovir, chemical name: [[(1S)-2-(4-amino-2-oxo-1(2H)-pyrimidinyl)-1-(hydroxymethyl)ethoxy]methyl]phosphonic acid mono[3-(hexadecyloxy)propyl] ester, is a broad-spectrum antiviral drug with anti-double-stranded DNA (dsDNA) virus activity. It is a nucleotide analogue, effective orally, and is a phosphoesterified prodrug of cidofovir. Its chemical formula is C 27 H 52 N3O7P, due to the poor bioavailability of cidofovir and its serious toxic side effects, it can cause serious irreversible nephrotoxicity, making it difficult to be widely used. As a prodrug of cidofovir, brincidofovir is modified by phospholipidation to enable it to be degraded in cells to release the effective active ingredient cidofovir, thereby improving the activity of cidofovir and its oral bioavailability. Compared with cidofovir, brincidofovir has the advantages of being oral, less harmful to the kidneys, and safe. It has stronger in vitro and in vivo activity than CDV against certain herpes viruses, adenoviruses, and orthopoxviruses. It is an antiviral drug used to treat cytomegalovirus, adenovirus, smallpox, and Ebola virus infections. It is suitable for the treatment of human smallpox disease caused by smallpox virus in adult and pediatric patients (including newborns). On June 4, 2021, the U.S. Food and Drug Administration approved brincidofovir brincidofovir tablets and oral suspension for the treatment of smallpox. However, brincidofovir has poor solubility, which limits its application in the formulation process.

[0003] Therefore, in view of the above problems, it is necessary to develop a brincidofovir trihydrate with good stability and solubility, low hygroscopicity, and convenient preparation of pharmaceutical preparations. Summary of the invention

[0004] The present invention aims to provide a brincidofovir compound and a preparation method thereof. The compound is brincidofovir trihydrate, has good stability and solubility, low hygroscopicity, is convenient for the preparation of pharmaceutical preparations, and improves the safety of medication.

[0005] In order to achieve the above object, the present invention adopts the following technical solution:

[0006] In the first aspect of the present invention, a brincidofovir compound is provided, wherein the brincidofovir compound is brincidofovir trihydrate, and the molecular formula is C 27 H 52N3O7P·3H2O.

[0007] Furthermore, the brincidofovir compound is crystalline and has an absorption peak between 130 and 135°C.

[0008] In a second aspect of the present invention, there is provided a method for preparing brincidofovir trihydrate, the method comprising:

[0009] S1. Add 0.05-0.2 mol / L sodium hydroxide solution and brincidofovir to a reaction pot in sequence, start stirring, heat to 40-44° C., and stir to dissolve; wherein the weight ratio of brincidofovir to 0.1 mol / L sodium hydroxide solution is 1:(14-16);

[0010] S2, cooling to 20-24°C, slowly adding a mixed solution of ethanol and chloroform while stirring, and continuing to stir after adding to obtain a material; wherein the amount of the mixed solution added is 7-9 times the weight of the sodium hydroxide solution in S1; the weight ratio of the mixed solution of ethanol and chloroform is: ethanol: chloroform = (8-10): 1

[0011] S3, transferring the material obtained in S2 to a crystallization kettle, cooling the temperature to 3-5°C at 3-3.6°C / min, filtering to obtain a filter cake, and washing the filter cake with purified water;

[0012] S4, drying the filter cake obtained in S3 and then crushing the filter cake to obtain brincidofovir trihydrate.

[0013] Furthermore, in S1, the weight ratio of brincidofovir to 0.1 mol / L sodium hydroxide solution is 1:15.

[0014] Furthermore, in S2, the weight ratio of the mixed solution of ethanol and chloroform is: ethanol:chloroform=9:1.

[0015] Furthermore, in S2, the stirring speed is maintained at 120-140 rpm.

[0016] Furthermore, in S3, during the rinsing, the filter cake is rinsed 1-3 times with 2-4 times the weight of purified water.

[0017] Furthermore, in S3, the drying condition is: drying at 60-66° C. for 6-8 hours.

[0018] In the third aspect of the present invention, a brincidofovir preparation is provided, wherein the preparation contains the brincidofovir compound.

[0019] One or more technical solutions in the embodiments of the present invention have at least the following technical effects or advantages:

[0020] 1. The present invention provides a brincidofovir trihydrate and a preparation method thereof. The inventors unexpectedly obtained a new brincidofovir trihydrate and a preparation method thereof during a long period of extensive research on brincidofovir. The brincidofovir trihydrate prepared by the method has extremely high quality stability, reduced hygroscopicity, and improved solubility, which is significantly superior to the prior art.

[0021] 2. In the preparation method of brincidofovir trihydrate provided by the present invention, the ratio of brincidofovir, 0.1 mol / L sodium hydroxide solution, ethanol, and chloroform, the reaction temperature, the cooling rate, and the time are crucial to obtaining the brincidofovir compound of the present invention. According to a large number of experiments by the applicant, the brincidofovir compound described in the present invention cannot be obtained by changing the relevant parameters of the present application. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the accompanying drawings required for use in the description of the embodiments will be briefly introduced below. Obviously, the accompanying drawings in the following description are some embodiments of the present invention. For ordinary technicians in this field, other accompanying drawings can be obtained based on these accompanying drawings without paying any creative work.

[0023] Figure 1 This is the X-ray powder diffraction pattern of the brincidofovir compound prepared in Example 1. DETAILED DESCRIPTION

[0024] The present invention will be described in detail below in conjunction with specific implementations and examples, and the advantages and various effects of the present invention will be more clearly presented. It should be understood by those skilled in the art that these specific implementations and examples are used to illustrate the present invention, rather than to limit the present invention.

[0025] Throughout the specification, unless otherwise specifically stated, the terms used herein should be understood as meanings commonly used in the art. Therefore, unless otherwise defined, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art to which the present invention belongs. In the event of a conflict, the present specification takes precedence.

[0026] Unless otherwise specified, various raw materials, reagents, instruments and equipment used in the present invention can be purchased from the market or obtained by existing methods.

[0027] The brincidofovir used can be synthesized according to known techniques, or purchased as a commercial product.

[0028] The present application will be described in detail below with reference to embodiments and experimental data.

[0029] Example 1. Preparation of Brincidofovir Compound

[0030] (1) 15 kg of 0.1 mol / L sodium hydroxide solution and 1 kg of brincidofovir were added to a reaction pot in sequence, stirred, heated to 40° C., and stirred to dissolve;

[0031] (2) cooling to 20° C., maintaining a stirring speed of 120 rpm, slowly adding 120 kg of a mixed solution of ethanol and chloroform while stirring, and continuing to stir for 30 minutes after the addition; the weight ratio of the mixed solution of ethanol and chloroform is: ethanol:chloroform = 9:1;

[0032] (3) transferring the material of step (2) to a crystallization kettle, cooling to 3°C at 3°C / min, filtering to obtain a filter cake, and washing the filter cake twice with 3 times the weight of purified water;

[0033] (4) placing the filter cake obtained in step (3) in a drying oven at 60° C., drying for 7 hours, and then crushing the filter cake to obtain the brincidofovir compound.

[0034] X-ray powder diffraction pattern is attached Figure 1 .

[0035] Example 2 Preparation of Brincidofovir Compound

[0036] (1) 15 kg of 0.1 mol / L sodium hydroxide solution and 1 kg of brincidofovir were added to a reaction pot in sequence, stirred, heated to 44° C., and stirred to dissolve;

[0037] (2) cooling to 24° C., maintaining a stirring speed of 140 rpm, slowly adding 120 kg of a mixed solution of ethanol and chloroform while stirring, and continuing to stir for 30 minutes after the addition; the weight ratio of the mixed solution of ethanol and chloroform is: ethanol:chloroform = 9:1;

[0038] (3) transferring the material of step (2) to a crystallization kettle, cooling the temperature to 5°C at 3.6°C / min, filtering to obtain a filter cake, and washing the filter cake twice with 3 times the weight of purified water;

[0039] (4) placing the filter cake obtained in step (3) in a drying oven at 66° C., drying for 7 hours, and then crushing the filter cake to obtain the brincidofovir compound.

[0040] The X-ray powder diffraction pattern was consistent with that of Example 1.

[0041] Example 3 Preparation of Brincidofovir Compound

[0042] (1) 15 kg of 0.1 mol / L sodium hydroxide solution and 1 kg of brincidofovir were added to a reaction pot in sequence, stirred, heated to 42° C., and stirred to dissolve;

[0043] (2) cooling to 22° C., maintaining a stirring speed of 130 rpm, slowly adding 120 kg of a mixed solution of ethanol and chloroform while stirring, and continuing to stir for 30 minutes after the addition; the weight ratio of the mixed solution of ethanol and chloroform is: ethanol: chloroform = 9:1;

[0044] (3) transferring the material of step (2) to a crystallization kettle, cooling the temperature to 4°C at 3.3°C / min, filtering to obtain a filter cake, and washing the filter cake twice with 3 times the weight of purified water;

[0045] (4) placing the filter cake obtained in step (3) in a drying oven at 63° C., drying for 7 hours, and then crushing the filter cake to obtain the brincidofovir compound.

[0046] The X-ray powder diffraction pattern was consistent with that of Example 1.

[0047] Comparative Example 1

[0048] (1) Add 15 kg of 0.1 mol / L sodium hydroxide solution and 1 kg of brincidofovir into a reaction pot in sequence, start stirring, heat to 40° C., and stir to dissolve;

[0049] (2) cooling to 20° C., maintaining a stirring speed of 120 rpm, slowly adding 120 kg of a mixed solution of ethanol and chloroform while stirring, and continuing to stir for 30 minutes after the addition; the weight ratio of the mixed solution of ethanol and chloroform is: ethanol: chloroform = 8:1;

[0050] (3) transferring the material of step (2) to a crystallization kettle, cooling to 3°C at 3°C / min, filtering to obtain a filter cake, and washing the filter cake twice with 3 times the weight of purified water;

[0051] (4) placing the filter cake obtained in step (3) in a drying oven at 60° C., drying for 7 hours, and then crushing the filter cake to obtain the brincidofovir compound.

[0052] Comparative Example 2

[0053] (1) Add 15 kg of 0.1 mol / L sodium hydroxide solution and 1 kg of brincidofovir into a reaction pot in sequence, start stirring, heat to 40° C., and stir to dissolve;

[0054] (2) cooling to 22° C., maintaining a stirring speed of 150 rpm, slowly adding 120 kg of a mixed solution of ethanol and chloroform while stirring, and continuing to stir for 30 minutes after the addition; the weight ratio of the mixed solution of ethanol and chloroform is: ethanol:chloroform = 9:1;

[0055] (3) transferring the material of step (2) to a crystallization kettle, cooling the temperature to 6°C at 4°C / min, filtering to obtain a filter cake, and washing the filter cake twice with 3 times the weight of purified water;

[0056] (4) placing the filter cake obtained in step (3) in a drying oven at 60° C., drying for 7 hours, and then crushing the filter cake to obtain the brincidofovir compound.

[0057] Comparative Example 3

[0058] (1) Add 15 kg of 0.1 mol / L sodium hydroxide solution and 1 kg of brincidofovir into a reaction pot in sequence, start stirring, heat to 40° C., and stir to dissolve;

[0059] (2) cooling to 20° C., maintaining a stirring speed of 110 rpm, slowly adding 110 kg of a mixed solution of ethanol and chloroform while stirring, and continuing to stir for 30 minutes after the addition; the weight ratio of the mixed solution of ethanol and chloroform is: ethanol:chloroform = 9:1;

[0060] (3) transferring the material of step (2) to a crystallization kettle, cooling to 3°C at 3°C / min, filtering to obtain a filter cake, and washing the filter cake twice with 3 times the weight of purified water;

[0061] (4) placing the filter cake obtained in step (3) in a drying oven at 60° C., drying for 7 hours, and then crushing the filter cake to obtain the brincidofovir compound.

[0062] Experimental Example 1: Study on the properties of brincidofovir trihydrate

[0063] 1. Elemental analysis

[0064] The brincidofovir trihydrate prepared by the method of the embodiment of the present invention was subjected to elemental analysis, and the results were: C: 52.66%, H: 9.45%, N: 6.85%, P: 5.02% (accurate to two decimal places), which is consistent with the theoretical value of brincidofovir trihydrate. The theoretical value of brincidofovir trihydrate is: C: 52.68%, H: 9.43%, N: 6.82%, P: 5.04% (accurate to two decimal places).

[0065] 2. Differential thermal analysis

[0066] The brincidofovir trihydrate prepared by the method of the present invention was subjected to differential thermal analysis and compared with brincidofovir trihydrate preparation raw material brincidofovir. The results showed that brincidofovir trihydrate had an absorption peak between 130 and 135°C, indicating that there was crystal water or crystallization solvent in the sample, while the reference substance had no absorption peak, indicating that there was no crystal water or crystallization solvent in the sample.

[0067] 3. Moisture analysis

[0068] The water content of the brincidofovir trihydrate of the present invention was measured by Karl Fischer water determination method, and the result was between 8.75% and 8.80%, which was consistent with the theoretical water content of brincidofovir trihydrate of 8.78%, and the two results were consistent, indicating that the sample contained only water and no other solvents.

[0069] Comprehensive elemental analysis, differential thermal analysis, and moisture analysis results can prove that brincidofovir of the present invention contains 3 molecules of crystal water.

[0070] 4. Experimental comparative analysis

[0071] Sample 1: the brincidofovir compound prepared in Example 1 of the present invention;

[0072] Sample 2: the brincidofovir raw material of Comparative Example 1;

[0073] 4.1 Characteristic Study:

[0074] The characteristics of samples 1-2 were studied according to the test method of this application, and the results are shown in Table 1.

[0075] Table 1 Results of the study on the properties of brincidofovir

[0076] sample characteristic Sample 1 Trihydrate Sample 2 No crystal water

[0077] 4.2 Stability and high humidity test:

[0078] Samples 1-2 were subjected to an influencing factor test and placed under high temperature (60°C), high humidity (RH92.5%±5%), and light (4500lx) conditions for 10 days. Samples were taken on the 10th day to examine the content and moisture content. The results were compared with those on day 0, see Table 2.

[0079] Table 2 Test results of factors affecting brincidofovir trihydrate

[0080]

[0081] The test results in Table 2 show that brincidofovir trihydrate has good stability and low hygroscopicity. The same test was carried out in other embodiments of the present invention and similar results were obtained.

[0082] 4.3 Solubility analysis

[0083] The solubility of samples 1-2 in water (25°C ± 2°C) is compared, and the results are shown in Table 3:

[0084] Table 3 Solubility comparison

[0085] sample Solubility (μg / ml) Sample 1 232 Sample 2 81

[0086] The test results in Table 3 show that the solubility of the brincidofovir compound prepared by the present invention is significantly improved. The same test was also carried out in other embodiments of the present invention, and similar results were obtained.

[0087] It can be seen from Table 2-3 that the brincidofovir trihydrate prepared by the present invention has better solubility and lower hygroscopicity than the brincidofovir anhydrous compound; in general, the solubility and dissolution rate of drugs in water are in the order of hydrate < anhydrous < organic solvate, and compounds with higher hygroscopicity have better solubility. The brincidofovir compound prepared by the present invention has achieved unexpected technical effects.

[0088] Test Example 2: Comparison of groups

[0089] The brincidofovir compounds prepared in Examples 1-3 of the present invention and Comparative Examples 1-3 were investigated for properties, hygroscopicity and solubility in water. The results are shown in Tables 4-6.

[0090] Table 4 Characteristic study

[0091] sample characteristic Example 1 Trihydrate Example 2 Trihydrate Example 3 Trihydrate Comparative Example 1 No crystal water Comparative Example 2 No crystal water Comparative Example 3 No crystal water

[0092] Table 5 Moisture absorption test (RH 92.5% ± 5% for 10 days)

[0093]

[0094] Table 6 Solubility comparison (25℃±2℃)

[0095] sample Solubility (μg / ml) Example 1 232 Example 2 231 Example 3 232 Comparative Example 1 79 Comparative Example 2 80 Comparative Example 3 82

[0096] It can be seen from the results in Tables 4-6 that the brincidofovir compound prepared by the present invention has low hygroscopicity and good solubility, and has obvious advantages over the prior art. The process parameters for preparing the brincidofovir compound of the present invention are crucial, and even a slight change will not result in the brincidofovir compound of the present invention.

[0097] Finally, it should be noted that the terms "comprises," "includes," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that includes a series of elements includes not only those elements, but also other elements not explicitly listed, or also includes elements that are inherent to such process, method, article, or apparatus.

[0098] Although the preferred embodiments of the present invention have been described, those skilled in the art may make other changes and modifications to these embodiments once they have learned the basic creative concept. Therefore, the appended claims are intended to be interpreted as including the preferred embodiments and all changes and modifications that fall within the scope of the present invention.

[0099] Obviously, those skilled in the art can make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include these modifications and variations.

Claims

1. A brincidofovir compound, characterized in that The brincidofovir compound is brincidofovir trihydrate, and the molecular formula is C 27 H 52 N3O7P·3H2O.

2. The brincidofovir compound according to claim 1, characterized in that The brincidofovir compound is crystalline and has an absorption peak between 130 and 135°C.

3. A method for preparing a brincidofovir compound according to claim 1 or 2, characterized in that: The method comprises: S1. Add 0.1 mol / L sodium hydroxide solution and brincidofovir to the reaction pot in sequence, start stirring, heat to 40-44° C., and stir to dissolve; wherein the weight ratio of brincidofovir to sodium hydroxide solution is 1:(14-16); S2, cooling to 20-24°C, slowly adding a mixed solution of ethanol and chloroform while stirring, and continuing to stir after the addition to obtain a material; wherein the amount of the mixed solution added is 7-9 times the weight of the sodium hydroxide solution in S1; the weight ratio of the mixed solution of ethanol and chloroform is ethanol: chloroform = (8-10): 1; S3, transferring the material obtained in S2 to a crystallization kettle, cooling the temperature to 3-5°C at 3-3.6°C / min, filtering to obtain a filter cake, and washing the filter cake with purified water; S4, drying the filter cake obtained in S3 and then crushing the filter cake to obtain brincidofovir trihydrate.

4. The preparation method according to claim 3, characterized in that: In the S1, the weight ratio of brincidofovir to 0.1 mol / L sodium hydroxide solution is 1:

15.

5. The preparation method according to claim 3, characterized in that: In S2, the weight ratio of the mixed solution of ethanol and chloroform is ethanol:chloroform=9:

1.

6. The preparation method according to claim 3, characterized in that: In S2, the stirring speed is maintained at 120-140 rpm.

7. The preparation method according to claim 3, characterized in that: In S3, during the washing, the filter cake is washed 1-3 times with 2-4 times the weight of purified water.

8. The preparation method according to claim 3, characterized in that: In S3, the drying condition is: drying at 60-66° C. for 6-8 hours.

9. A brincidofovir preparation, characterized in that The preparation contains the brincidofovir compound according to claim 1 or 2.

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