A brincidofovir compound and methods of making the same

Abstract

The application provides a brincidofovir compound and a preparation method thereof, and the method comprises the following steps: sequentially adding 0.05-0.2 mol / L of a sodium hydroxide solution and brincidofovir into a reaction kettle, starting stirring, heating to 40-44 DEG C, and stirring and dissolving; cooling to 20-24 DEG C, slowly adding a mixed solution of ethanol and chloroform while stirring, continuing to stir after the addition is completed, and obtaining a material; then transferring the material to a crystallization kettle, cooling to 3-5 DEG C at a cooling rate of 3-3.6 DEG C / min, filtering to obtain a filter cake, crushing the filter cake after drying, and obtaining brincidofovir trihydrate. The inventor of the application has unexpectedly obtained a new brincidofovir compound and a preparation method thereof in the process of long-term and large amount of research on brincidofovir. The brincidofovir compound prepared by the method has extremely high quality stability, and the hygroscopicity is reduced and the solubility is improved.

Description

TECHNICAL FIELD

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

[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, is a nucleotide analog, is orally effective, is a prodrug of cidofovir phosphate, and has the chemical formula C 27 H 52 N3O7P, due to the poor bioavailability of cidofovir and the serious toxic side effects, can cause serious irreversible nephrotoxicity, so that it is difficult to be widely used. Brincidofovir, as a prodrug of cidofovir, is modified by phosphatidation to allow it to be degraded in cells to release the active ingredient cidofovir, thereby improving the activity of cidofovir and improving its oral bioavailability. Compared with cidofovir, brincidofovir has the advantages of being orally available, less harmful to the kidneys, safe, etc. It has stronger in-vivo and in-vitro activity than CDV against some herpes viruses, adenoviruses and orthopoxviruses, and is an antiviral drug for treating cytomegalovirus, adenovirus, smallpox and Ebola virus infections. It is suitable for treating human smallpox disease caused by smallpox virus in adult and pediatric patients (including neonates). On June 4, 2021, the U.S. Food and Drug Administration approved brincidofovir tablets and oral suspensions of brincidofovir for the treatment of smallpox. However, the poor solubility of brincidofovir limits its use in the preparation process.

[0003] Therefore, in view of the above problems, it is necessary to develop a brincidofovir trihydrate which has good stability and solubility, low hygroscopicity, and facilitates the preparation of drug preparations. SUMMARY

[0004] The present application aims to provide a brincidofovir compound and a preparation method thereof, which is brincidofovir trihydrate, has good stability and solubility, low hygroscopicity, facilitates the preparation of drug preparations, and improves the safety of medication.

[0005] In order to achieve the above-mentioned purpose, the present application adopts the following technical solutions:

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

[0007] Further, the brincidofovir compound is a crystal, and has an absorption peak between 130 and 135 DEG C.

[0008] In a second aspect of the present application, a preparation method of brincidofovir trihydrate is provided, the method comprising:

[0009] S1, a 0.05-0.2 mol / L sodium hydroxide solution and brincidofovir are sequentially added into a reaction kettle, stirring is started, and heating is performed to 40-44 DEG C for dissolution; wherein the brincidofovir and the 0.1 mol / L sodium hydroxide solution are used in a weight ratio of 1:(14-16);

[0010] S2, cooling is performed to 20-24 DEG C, a mixed solution of ethanol and chloroform is slowly added while stirring, and after the addition is completed, stirring is continued to obtain a material; wherein the mixed solution is added in an amount of 7-9 times the weight of the sodium hydroxide solution in S1; and the mixed solution of ethanol and chloroform is used in a weight ratio of ethanol: chloroform = (8-10):1.

[0011] S3, the material obtained in S2 is transferred to a crystallization kettle, cooling is performed to 3-5 DEG C at a rate of 3-3.6 DEG C / min, filtration is performed to obtain a filter cake, and the filter cake is washed with purified water;

[0012] S4, the filter cake obtained in S3 is dried, the filter cake is crushed to obtain brincidofovir trihydrate.

[0013] Further, in S1, the brincidofovir and the 0.1 mol / L sodium hydroxide solution are used in a weight ratio of 1:15.

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

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

[0016] Further, in S3, in the washing, the filter cake is washed with 2-4 times the weight of purified water for 1-3 times.

[0017] Further, in S3, the drying condition is that drying is performed at 60-66 DEG C for 6-8 hours.

[0018] In a third aspect of the present application, a brincidofovir preparation is provided, the preparation containing the brincidofovir compound.

[0019] The one or more technical solutions in the embodiments of the present application have at least the following technical effects or advantages:

[0020] 1.The brincidofovir trihydrate and the preparation method thereof, the brincidofovir trihydrate and the preparation method thereof are obtained accidentally by the inventors in the long-term research on brincidofovir, the brincidofovir trihydrate prepared by the method has extremely high quality stability, and the hygroscopicity is reduced, the solubility is improved, and the brincidofovir trihydrate is obviously superior to the prior art.

[0021] 2.In the preparation method of the brincidofovir trihydrate, the ratio of brincidofovir, 0.1mol / L sodium hydroxide solution, ethanol and chloroform, the reaction temperature, the cooling rate and the time are crucial to obtain the brincidofovir compound of the application, according to a large number of tests of the applicant, the brincidofovir compound of the application cannot be obtained by changing the related parameters of the application. BRIEF DESCRIPTION OF DRAWINGS

[0022] In order to more clearly illustrate the technical solutions in the embodiments of the application, the drawings needed in the embodiment description will be briefly introduced. Obviously, the drawings in the following description are some embodiments of the application, and other drawings can be obtained by those skilled in the art without creative labor.

[0023] Figure 1 X-ray powder diffraction pattern of the brincidofovir compound prepared for example 1. DETAILED DESCRIPTION

[0024] The advantages and various effects of the application will be more clearly presented by the following specific embodiments and examples. Those skilled in the art should understand that these specific embodiments and examples are used to illustrate the application, not to limit the application.

[0025] Throughout the specification, unless otherwise specifically indicated, the terms used herein are understood as having the meanings commonly used in the art. Therefore, unless otherwise defined, all technical and scientific terms used herein have the same meaning as generally understood by those skilled in the art to which the application belongs. If there is a contradiction, the specification takes precedence.

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

[0027] The brincidofovir used can be synthesized according to known technology, or can be purchased commercially.

[0028] The application will be described in detail below in combination with examples and experimental data.

[0029] Example 1, Preparation of Brincidofovir compound

[0030] (1) 15 kg of 0.1 mol / L sodium hydroxide solution, 1 kg of Brincidofovir were added into the reaction pot in turn, the stirring was started, and heated to 40℃, and dissolved by stirring;

[0031] (2) The temperature was lowered to 20℃, the stirring speed was kept at 120 rpm, 120 kg of mixed solution of ethanol and chloroform was slowly added while stirring, and after the addition was completed, the stirring was continued for 30 minutes; the amount of the mixed solution of ethanol and chloroform was ethanol: chloroform = 9:1 by weight;

[0032] (3) The material of step (2) was transferred to a crystallization kettle, and cooled to 3℃ at a rate of 3℃ / min, filtered to obtain a filter cake, and the filter cake was washed with 3 times the weight of purified water for 2 times;

[0033] (4) The filter cake obtained in step (3) was placed in a drying box, dried at 60℃ for 7 hours, and then the filter cake was crushed to obtain the Brincidofovir compound.

[0034] The X-ray powder diffraction pattern is shown in the attached Figure 1 .

[0035] Example 2, Preparation of Brincidofovir compound

[0036] (1) 15 kg of 0.1 mol / L sodium hydroxide solution, 1 kg of Brincidofovir were added into the reaction pot in turn, the stirring was started, and heated to 44℃, and dissolved by stirring;

[0037] (2) The temperature was lowered to 24℃, the stirring speed was kept at 140 rpm, 120 kg of mixed solution of ethanol and chloroform was slowly added while stirring, and after the addition was completed, the stirring was continued for 30 minutes; the amount of the mixed solution of ethanol and chloroform was ethanol: chloroform = 9:1 by weight;

[0038] (3) The material of step (2) was transferred to a crystallization kettle, and cooled to 5℃ at a rate of 3.6℃ / min, filtered to obtain a filter cake, and the filter cake was washed with 3 times the weight of purified water for 2 times;

[0039] (4) The filter cake obtained in step (3) was placed in a drying box, dried at 66℃ for 7 hours, and then the filter cake was crushed to obtain the Brincidofovir compound.

[0040] The X-ray powder diffraction pattern is 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, 1 kg of brincidofovir were sequentially added into a reaction kettle, stirring was started, and heating was performed to 42°C, and dissolution was performed under stirring;

[0043] (2) The temperature was lowered to 22°C, the stirring speed was kept at 130 rpm, 120 kg of a mixed solution of ethanol and chloroform was slowly added under stirring, and stirring was continued for 30 minutes after the addition was completed; the mixed solution of ethanol and chloroform was used in a weight ratio of ethanol: chloroform = 9: 1;

[0044] (3) The material of step (2) was transferred to a crystallization kettle, the temperature was lowered to 4°C at a rate of 3.3°C / min, and filtration was performed, and a filter cake was obtained, and the filter cake was washed twice with 3 times the weight of purified water;

[0045] (4) The filter cake obtained in step (3) was placed in a drying box, dried at 63°C for 7 hours, and then the filter cake was crushed to obtain a brincidofovir compound.

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

[0047] Comparative Example 1

[0048] (1) 15 kg of 0.1 mol / L sodium hydroxide solution, 1 kg of brincidofovir were sequentially added into a reaction kettle, stirring was started, and heating was performed to 42°C, and dissolution was performed under stirring;

[0049] (2) The temperature was lowered to 22°C, the stirring speed was kept at 130 rpm, 120 kg of a mixed solution of ethanol and chloroform was slowly added under stirring, and stirring was continued for 30 minutes after the addition was completed; the mixed solution of ethanol and chloroform was used in a weight ratio of ethanol: chloroform = 9: 1;

[0050] (3) The material of step (2) was transferred to a crystallization kettle, the temperature was lowered to 4°C at a rate of 3.3°C / min, and filtration was performed, and a filter cake was obtained, and the filter cake was washed twice with 3 times the weight of purified water;

[0051] (4) The filter cake obtained in step (3) was placed in a drying box, dried at 63°C for 7 hours, and then the filter cake was crushed to obtain a brincidofovir compound.

[0052] Comparative Example 2

[0053] (1) 15 kg of 0.1 mol / L sodium hydroxide solution, 1 kg of brincidofovir were sequentially added into a reaction kettle, stirring was started, and heating was performed to 42°C, and dissolution was performed under stirring;

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

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

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

[0057] Comparative Example 3

[0058] (1) sequentially adding 15 kg of a 0.1 mol / L sodium hydroxide solution and 1 kg of brincidofovir to a reaction kettle, turning on the stirring, and heating to 40℃ and dissolving while stirring;

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

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

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

[0062] Experimental Example 1, Characteristic Study of Brincidofovir Trihydrate

[0063] 1. Elemental Analysis

[0064] The brincidofovir trihydrate prepared by the method of the present application 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 was consistent with the theoretical value of the brincidofovir trihydrate, and the theoretical value of the brincidofovir trihydrate was: 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 application is subjected to differential thermal analysis, and compared with the brincidofovir trihydrate raw material brincidofovir. The results show that the brincidofovir trihydrate has an absorption peak between 130-135°C, indicating that the sample contains crystalline water or crystalline solvent, while the control sample has no absorption peak, indicating that the sample contains no crystalline water or crystalline solvent.

[0067] 3. Moisture analysis

[0068] The moisture content of the brincidofovir trihydrate described in the present application is determined according to the Karl Fischer moisture titration method, and the result is between 8.75-8.80%, which is consistent with the theoretical water content of the brincidofovir trihydrate, 8.78%. This shows that the sample only contains water, and no other solvent.

[0069] Based on the results of elemental analysis, differential thermal analysis, and moisture analysis, it can be proved that the brincidofovir of the present application contains 3 molecules of crystalline water.

[0070] 4. Comparative analysis of tests

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

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

[0073] 4.1 Characteristic study:

[0074] The samples 1-2 are subjected to characteristic study according to the test method of the present application, and the results are shown in Table 1.

[0075] Table 1: Results of brincidofovir characteristic study

[0076] Sample Properties Sample 1 Trihydrate Sample 2 Without crystallization water

[0077] 4.2 Stability and moisture absorption test under high humidity:

[0078] The samples 1-2 are subjected to influence factor test to investigate the content and moisture after being placed under high temperature (60°C), high humidity (RH 92.5%±5%), and light (4500lx) for 10 days, respectively. The results are compared with those at 0 days, and are shown in Table 2.

[0079] Table 2: Results of brincidofovir trihydrate influence factor test

[0080]

[0081] The test results in Table 2 show that the brincidofovir trihydrate has good stability and low moisture absorption. The same test is also conducted on other examples of the present application, and similar results are obtained.

[0082] 4.3 Solubility analysis

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

[0084] Table 3 Solubility comparison

[0085] Sample Solubility (pg / 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 application is significantly improved. The same test was also performed on other embodiments of the present application, and similar results were obtained.

[0087] As can be seen from Tables 2-3, the brincidofovir trihydrate prepared by the present application has better solubility and lower hygroscopicity than the brincidofovir anhydrous compound. Generally, the solubility and dissolution rate of drugs in water follow the order: hydrate < anhydrous < organic solvate, and compounds with higher hygroscopicity have better solubility. The brincidofovir compound prepared by the present application achieves unexpected technical effects.

[0088] Test Example 2: Comparison of each group

[0089] The brincidofovir compounds prepared by the present application examples 1-3 and comparative examples 1-3 were subjected to property research, hygroscopicity and water solubility investigation, and the results are shown in Tables 4-6;

[0090] Table 4 Property research

[0091] Sample Properties Example 1 Trihydrate Example 2 Trihydrate Example 3 Trihydrate Comparative Example 1 Without crystallization water Comparative Example 2 Without crystallization water Comparative Example 3 Without crystallization water

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

[0093]

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

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

[0096] As can be seen from the results in Tables 4-6, the brincidofovir compound prepared by the present application has low hygroscopicity and good solubility, and has obvious advantages compared with the prior art. The process parameters for preparing the brincidofovir compound of the present application are crucial, and slight changes cannot obtain the brincidofovir compound described in the present application.

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

[0098] While the preferred embodiments of the application have been described, additional variations and modifications can be made to the preferred embodiments by those of skill in the art once they have the benefit of the present disclosure without departing from the spirit and scope of the application. Accordingly, it is intended that the appended claims encompass all such variations and modifications as falling within the scope of the application.

[0099] Obviously, numerous modifications and variations of the present application are possible in light of the above teachings. It is therefore to be understood that within the scope of the appended claims and their equivalents, the application can be practiced otherwise than as specifically described.

Claims

1. A brincidofovir compound, characterized by, The brincidofovir compound is brincidofovir trihydrate having the formula: 27 H 52 N3O7P-3H2O, the brincidofovir compound is crystalline and has an absorption peak between 130 and 135 °C, and the crystal has an X-ray powder diffraction pattern as shown in Figure 1.

2. The method for preparing a brincidofovir compound according to claim 1, characterized in that, The method comprises: S1, 0.1 mol / L of sodium hydroxide solution, Brincidofovir is added to the reaction pot in turn, open the stirring, heating to 40~44℃, stirring dissolution; wherein, the Brincidofovir, sodium hydroxide solution dosage weight ratio is 1: (14-16) ; S2, cooling to 20~24℃, slowly add the mixed solution of ethanol and chloroform while stirring, continue to stir after adding, get material; wherein, the mixed solution is 7~9 times of the weight of sodium hydroxide solution in S1; the mixed solution of ethanol and chloroform is ethanol: chloroform = 9: 1; in S2, the stirring speed is kept at 120~140 r / min; S3, the material obtained in S2 is transferred to the crystallization kettle, and the temperature is reduced to 3~5℃ at a rate of 3~3.6℃ / min, and then filtered to obtain a filter cake, which is washed with purified water; S4, the filter cake obtained in S3 is dried, and then the filter cake is crushed to obtain Brincidofovir trihydrate.

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

4. The production method according to claim 2, characterized by, In S4, the drying conditions are: drying at 60~66℃ for 6-8 hours.

5. A brincidofovir formulation, characterized in that, The preparation contains the Brincidofovir compound of claim 1.

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