Sodium deoxycholate crystal form and preparation method and application thereof

Abstract

The invention relates to a sodium deoxycholate crystal form and a preparation method and application thereof, the sodium deoxycholate crystal form is selected from any one of a crystal form A and a crystal form B, the crystal form A has an X-ray powder diffraction pattern basically the same as that in a figure 1, and the crystal form B has an X-ray powder diffraction pattern basically the same as that in a figure 2. In addition, the invention also discloses a preparation method and application of the sodium deoxycholate crystal form.

Description

Technical Field

[0001] The present invention relates to, but is not limited to, the technical field of drug crystal forms, and in particular to a crystal form of sodium deoxycholate, a preparation method thereof and an application thereof. Background Art

[0002] Polymorphism refers to the ability of a compound to assume at least two crystalline arrangements in the solid state. In the pharmaceutical industry, the polymorphic form of a compound is of great interest because polymorphism can affect the solubility and stability of the compound. The study of polymorphism also helps reduce the risk and cost of problems during large-scale production.

[0003] Sodium Deoxycholate is the sodium salt of deoxycholic acid. It is a white or off-white powder at room temperature, with a bile-like odor and a strong bitter taste. Sodium Deoxycholate is an ionic detergent that can be used to lyse cells and dissolve proteins that are poorly soluble in water. Sodium Deoxycholate can also be used in bile lysis experiments. The principle is that bile or bile salts have surface activity, which can quickly activate autolytic enzymes and accelerate the self-lysis of bacteria such as Streptococcus pneumoniae. Sodium Deoxycholate is a pharmaceutical excipient included in the Chinese Pharmacopoeia and can be used as a surfactant and emulsifier.

[0004] The chemical name of sodium deoxycholate is: 3α,12α-dihydroxy-5β-cholestane-24-acid sodium, and the specific structural formula is as follows:

[0005] Summary of the invention

[0006] The inventors have found through investigation that the sodium deoxycholate currently sold on the market is amorphous and has a hygroscopicity problem, which is not conducive to better storage and industrial production of sodium deoxycholate.

[0007] The present application provides a crystal form of sodium deoxycholate and a preparation method thereof, which can be placed under conditions of high temperature, high humidity and light for a period of time, and both crystal forms A and B remain unchanged, and both have good stability and low hygroscopicity, which is conducive to better storage and industrial production of sodium deoxycholate.

[0008] In a first aspect, the present application provides a crystalline form of sodium deoxycholate represented by formula (I);

[0009]

[0010] The crystal form remains stable under high temperature, high humidity and light conditions.

[0011] In some embodiments, the crystalline form of sodium deoxycholate provided herein is sodium deoxycholate crystalline form A, which shows characteristic peaks at 5.1, 10.2, 11.6, 12.7, 13.6, 15.5, 18.6, 20.1 and 22.0 in its X-ray powder diffraction spectrum represented by a diffraction angle of 2θ±0.2°.

[0012] In some embodiments, the crystalline form of sodium deoxycholate provided herein is sodium deoxycholate crystalline form A, which shows characteristic peaks at 5.1, 10.2, 11.6, 12.7, 13.6, 15.5, 17.9, 18.6, 19.4, 20.1, 22.0 and 23.8 in its X-ray powder diffraction spectrum represented by a diffraction angle of 2θ±0.2°.

[0013] In some embodiments, the crystalline form of sodium deoxycholate provided in the present application is sodium deoxycholate crystalline form A, which has the same Figure 1 Essentially the same X-ray powder diffraction pattern.

[0014] In some embodiments, the crystalline form of sodium deoxycholate provided herein is sodium deoxycholate crystalline form A, which has a water content of 0.1% w / w to 5% w / w, preferably, 0.5% w / w to 4.9% w / w, or 0.7% w / w to 4.5% w / w, or 0.8% w / w to 4% w / w, or 0.8% w / w to 3.8% w / w.

[0015] In some embodiments, the crystalline form of sodium deoxycholate provided herein is sodium deoxycholate crystalline form B, which shows characteristic peaks at 7.1, 7.8, 9.4, 11.7, 13.0, 14.4, 15.6, 17.9, 21.1 and 23.2 in its X-ray powder diffraction spectrum represented by a diffraction angle of 2θ±0.2°.

[0016] In some embodiments, the crystalline form of sodium deoxycholate provided herein is sodium deoxycholate crystalline form B, which shows characteristic peaks at 7.1, 7.8, 9.4, 11.7, 13.0, 14.4, 15.6, 17.9, 19.0, 21.1, 21.6, 23.2 and 23.7 in its X-ray powder diffraction spectrum represented by a diffraction angle of 2θ±0.2°.

[0017] In some embodiments, the crystalline form of sodium deoxycholate provided in the present application is sodium deoxycholate crystalline form B, which has the same Figure 3 Essentially the same X-ray powder diffraction pattern.

[0018] In some embodiments, the crystalline form of sodium deoxycholate provided herein is sodium deoxycholate crystalline form B, which has a water content of 0.1% w / w to 3% w / w, preferably, 0.2% w / w to 3% w / w, or 0.1% w / w to 1% w / w, or 0.2% w / w to 1% w / w, or 0.2% w / w to 2% w / w.

[0019] Due to different measurement conditions, the 2θ angle and relative intensity of each peak on the X-ray powder diffraction pattern will vary. Generally, the 2θ angle changes within ± 0.2 and the relative intensity within ± 0.2% are considered to be reasonable errors. It should also be understood that the relative intensity of the peak may fluctuate with experimental conditions and sample preparation such as the preferred orientation of particles in the sample. The use of automatic or fixed divergence slits will also affect the calculation of relative intensity. The intensity shown in the XRPD curve included here is only exemplary and cannot be used as an absolute comparison.

[0020] In a second aspect, the present application provides a method for preparing the above-mentioned crystalline form of sodium deoxycholate, wherein the crystalline form is crystalline form A, and the preparation method comprises:

[0021] Mix deoxycholic acid and the first solvent, add sodium hydroxide, heat to 50°C to 65°C to dissolve, cool to 40°C to 55°C, filter to remove mechanical impurities; heat to 55°C to 65°C, add the second solvent, cool to 10°C to 30°C, crystallize for 1 to 3 hours, filter, and dry to obtain Form A;

[0022] The first solvent is selected from at least one of water, acetone, tetrahydrofuran, 1,4-dioxane, methanol, ethanol or a combination thereof; the second solvent is selected from at least one of water, acetone, tetrahydrofuran, 1,4-dioxane, methanol, ethanol or a combination thereof.

[0023] In some embodiments, the present application provides a method for preparing Form A of sodium deoxycholate, wherein the first solvent is water; the weight volume ratio of deoxycholic acid to water is (1-10):(10-1), expressed in g / ml; optionally, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4 or 1:5.

[0024] In some embodiments, the preparation method of Form A of sodium deoxycholate provided herein, wherein the first solvent is a mixed solution of water and acetone; in some examples, the volume ratio of water to acetone is (1-50):(50-1), optionally, the volume ratio of water to acetone is 3:1, 2:1, 1:1, 1:2, 1:3 or 1:4; in some examples, the weight volume ratio of deoxycholic acid to the mixed solution of water and acetone is (1-10):(10-1), the unit is g / ml; optionally, it is 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9 or 1:10.

[0025] In some embodiments, the preparation method of Form A of sodium deoxycholate provided herein, wherein the first solvent is a mixed solution of water and tetrahydrofuran; in some examples, the volume ratio of water to tetrahydrofuran is (1-50):(50-1), optionally, the volume ratio of water to tetrahydrofuran is 3:1, 2:1, 1:1, 1:2, 1:3 or 1:4; in some examples, the weight volume ratio of deoxycholic acid to the mixed solution of water and tetrahydrofuran is (1-10):(10-1), the unit is g / ml; optionally, it is 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7 or 1:8.

[0026] In some embodiments, the present application provides a method for preparing Form A of sodium deoxycholate, wherein the second solvent is acetone or tetrahydrofuran.

[0027] In some embodiments, the preparation method of Form A of sodium deoxycholate provided herein, wherein the volume ratio of the second solvent to the first solvent is (1-50):(50-1), optionally, 1:3, 1:2, 1:1, 7:6, 2:1, 5:1, 10:1, 15:1, 20:1, 25:1, 30:1, 35:1, 40:1, 45:1, or 50:1.

[0028] In some embodiments, the present application provides a method for preparing Form A of sodium deoxycholate, wherein the first solvent is water and the second solvent is acetone.

[0029] In some embodiments, the present application provides a method for preparing Form A of sodium deoxycholate, wherein the first solvent is a mixed solution of water and acetone, and the second solvent is acetone.

[0030] In some embodiments, the present application provides a method for preparing Form A of sodium deoxycholate, wherein the first solvent is a mixed solution of water and tetrahydrofuran, and the second solvent is tetrahydrofuran.

[0031] In a second aspect, the present application provides a method for preparing a crystalline form of sodium deoxycholate, wherein the crystalline form is crystalline form B, and the preparation method comprises:

[0032] The sodium deoxycholate crystal form A is mixed with the third solvent, and the obtained solution is concentrated under reduced pressure to obtain a solid, which is then dried to obtain the crystal form B.

[0033] In some embodiments, the preparation method of Form B of sodium deoxycholate provided herein, wherein the third solvent is selected from at least one of anhydrous methanol, anhydrous ethanol, isopropanol, n-propanol, n-butanol or a combination thereof.

[0034] In some embodiments, the present application provides a method for preparing Form B of sodium deoxycholate, wherein the third solvent is anhydrous ethanol.

[0035] In some embodiments, the present application provides a method for preparing Form B of sodium deoxycholate, wherein the third solvent is anhydrous methanol.

[0036] In some embodiments, the present application provides a method for preparing Form B of sodium deoxycholate, wherein the third solvent is isopropanol.

[0037] In some embodiments, the present application provides a method for preparing Form B of sodium deoxycholate, wherein the weight volume ratio of Form A of sodium deoxycholate to the third solvent is (10-1):(1-50), expressed in g / ml; optionally, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19, 1:20, 1:30 or 1:40.

[0038] In some embodiments, the present application provides a method for preparing Form B of sodium deoxycholate, wherein the sodium deoxycholate Form A can be prepared using the above-mentioned preparation method.

[0039] In a third aspect, the present application provides the use of the above-mentioned crystalline form of sodium deoxycholate as a detergent, emulsifier or pharmaceutical excipient, wherein the crystalline form is selected from any one of crystalline form A and crystalline form B.

[0040] Compared with the existing sodium deoxycholate, the present application provides a new crystal form of sodium deoxycholate and a preparation method thereof. After being placed under high temperature, high humidity and light conditions for a period of time, both crystal forms A and B remain unchanged in crystal form, and both have good stability and low hygroscopicity, which is conducive to better storage and industrial production of sodium deoxycholate.

[0041] Other features and advantages of the present application will be described in the following description, and partly become apparent from the description, or be understood by implementing the present application. Other advantages of the present application can be realized and obtained by the schemes described in the description and the drawings. BRIEF DESCRIPTION OF THE DRAWINGS

[0042] In order to more clearly illustrate the technical solutions of the embodiments of the present application, the drawings required for use in the description of the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present application. For ordinary technicians in this field, other embodiments can also be obtained based on these drawings.

[0043] Figure 1 is the X-ray powder diffraction pattern of sodium deoxycholate form A of Example 1;

[0044] Figure 2 is the X-ray powder diffraction pattern of sodium deoxycholate form A of Example 2;

[0045] Figure 3 is the X-ray powder diffraction pattern of sodium deoxycholate form B of Example 4;

[0046] Figure 4 This is the X-ray powder diffraction pattern of sodium deoxycholate form B of Example 5. DETAILED DESCRIPTION

[0047] The present invention will be further described below by specific examples, but the specific examples are not intended to limit the scope of protection of the present invention. Those skilled in the art may make improvements to the preparation method and the apparatus used within the scope of the claims, and these improvements should also be regarded as the scope of protection of the present invention. Therefore, the scope of protection of the patent of the present invention shall be subject to the attached claims.

[0048] The X-ray powder diffraction pattern of the present invention is collected on a Bruker D2 phaser X-ray powder diffractometer.

[0049] The method parameters of X-ray powder diffraction described in the present invention are as follows:

[0050] Voltage: 30KV

[0051] Current: 10mA

[0052] Scanning range: 3°~60°

[0053] Step size: 0.02°

[0054] Step speed: 0.5s / step

[0055] Example 1: Preparation of Form A

[0056] Add 460g of deoxycholic acid, 920ml of purified water and 920ml of acetone to the reaction bottle, stir and disperse evenly, and slowly add 43.6g of NaOH in batches. Heat to 50-60℃ to dissolve, turn off the heating, cool naturally to 45±5℃, and filter the membrane to remove mechanical impurities. The filtrate is transferred to the reaction bottle, heated to 60±5℃, and 3680ml of acetone is slowly added dropwise. After the addition is completed, keep warm at 60±5℃ and stir for 0.5h. Cool to 15±5℃ for crystallization for 1h, filter the filter cake and send it to a blast oven at 65℃ to dry 378g of product, with a yield of 78%, a purity of 99%, and a water content of 0.89% w / w (according to the drying loss method (Chinese Pharmacopoeia 2020 Edition Four General Rules 0831), the same below). The product was proved to be Form A of sodium deoxycholate by X-ray powder diffraction, see Attachment Figure 1 .

[0057] Example 2: Preparation of Form A

[0058] Add 10g of deoxycholic acid and 5ml of purified water to the reaction bottle, stir and disperse evenly, and slowly add 0.95g of NaOH in batches. Heat to 50-55℃ to dissolve, turn off the heating, cool naturally to 45±5℃, and filter to remove mechanical impurities. Transfer the filtrate to the reaction bottle, heat to 60±5℃, slowly add 200ml of acetone, and after the addition is complete, keep warm at 60±5℃ and stir for 0.5h. Cool to 15±5℃ for crystallization for 2h, filter the filter cake and send it to a forced air oven to dry at 65℃ to obtain 8g of product, with a yield of 76%, a purity of 98%, and a water content of 0.96% w / w. The product was confirmed to be Form A of sodium deoxycholate by X-ray powder diffraction. See attached Figure 2 .

[0059] Example 3: Preparation of Form A

[0060] Add 100g deoxycholic acid, 300ml purified water and 300ml tetrahydrofuran to the reaction bottle, stir and disperse evenly, and slowly add 9.5g NaOH in batches. Heat to 60-65℃ to dissolve, turn off the heating, cool naturally to 45-55℃, and filter to remove mechanical impurities. The filtrate is transferred to the reaction bottle, heated to 60±5℃, and 700ml tetrahydrofuran is slowly added dropwise. After the addition is completed, keep warm at 60±5℃ and stir for 0.5h. Cool to 10-15℃ for crystallization for 3h, filter the filter cake and send it to a blast oven at 65℃ to dry 85g of product, with a yield of 80%, a purity of 98%, and a water content of 1.20% w / w. The product was confirmed to be Form A of sodium deoxycholate by X-ray powder diffraction.

[0061] Example 4: Preparation of Form B

[0062] Add 5g of sodium deoxycholate (Crystal Form A of Example 1) to the reaction flask, stir and disperse with 60ml of anhydrous ethanol, heat to 50-60°C and concentrate to dryness under reduced pressure to obtain 5g of product with a yield of 100%, a purity of 99%, and a water content of 0.52% w / w. The product was confirmed to be Crystal Form B of sodium deoxycholate by X-ray powder diffraction. See attached Figure 3 .

[0063] Example 5: Preparation of Form B

[0064] 6 g of sodium deoxycholate (Crystal Form A of Example 2) was added to the reaction flask, and 60 ml of isopropanol was used for stirring and dispersion, and the temperature was raised to 60°C to 70°C, and the product was concentrated to dryness under reduced pressure to obtain 6 g of product with a yield of 100%, a purity of 98%, and a water content of 0.49% w / w. The product was confirmed to be Crystal Form B of sodium deoxycholate by X-ray powder diffraction. See attached Figure 4 .

[0065] Example 6: Preparation of Form B

[0066] 10 g of sodium deoxycholate (Crystal Form A of Example 3) was added to the reaction flask, dissolved in 100 ml of anhydrous methanol, heated to 45-55° C., and concentrated to dryness under reduced pressure to obtain 10 g of the product with a yield of 100%, a purity of 98%, and a water content of 0.63% w / w. The product was confirmed to be Crystal Form B of sodium deoxycholate by X-ray powder diffraction.

[0067] Example 7: Stability of Sodium Deoxycholate Crystalline Form

[0068] The stability of sodium deoxycholate crystalline form A and crystalline form B provided in the present application was investigated. Specifically, the changes in the crystalline form were investigated by placing each crystalline form under different storage conditions to measure the stability of the crystalline form to conditions such as high temperature, high humidity, and light. The storage conditions and results are shown in the following table:

[0069]

[0070] From the data in the table, it can be seen that sodium deoxycholate crystal form A and crystal form B can maintain good stability under high temperature, high humidity and light conditions. The X-ray diffraction patterns of crystal form A and crystal form B are consistent with the initial data, and no crystal transformation occurs, indicating that the crystal form provided by the present invention has good stability.

[0071] In order to further verify the chemical stability of Form A and Form B, the changes of related substances under different storage conditions were evaluated. The storage conditions and impurity contents are shown in the following table:

[0072]

[0073] The data in the table show that sodium deoxycholate crystal form A and crystal form B can maintain good chemical stability under the influencing factors, indicating that the crystal form provided by the present invention has good stability and is conducive to storage.

[0074] Example 8: Solubility study of sodium deoxycholate crystals

[0075] Take an appropriate amount of sodium deoxycholate (about 10-20 mg) and place it in a 25°C constant temperature mixer, slowly add the test solvent and shake, record the amount of solvent before and after dissolution, and calculate the solubility.

[0076] Dissolution conditions Crystalline Form A Crystalline Form B Amorphous (Sigma) water Soluble Soluble Soluble Anhydrous methanol Soluble Soluble Dissolve Anhydrous ethanol Soluble Soluble Dissolve

[0077] As can be seen from the above table, the sodium deoxycholate crystalline form A and crystalline form B provided in the present application show better solubility in various solutions than the amorphous form.

[0078] Example 9: Hygroscopicity of Sodium Oxycholate Crystalline Forms

[0079] Take an appropriate amount of sodium deoxycholate and place it in an open environment at a specific humidity for half an hour to test the moisture content of the quick-drying product.

[0080]

[0081] It can be seen from the above table that the hygroscopicity of sodium deoxycholate crystalline form A and crystalline form B provided in the present application is weaker than that of the amorphous form and has good hygroscopicity.

Claims

1. A crystalline form of sodium deoxycholate represented by formula (I); wherein the crystalline form of sodium deoxycholate remains stable under high temperature, high humidity and light conditions:

2. The crystal form according to claim 1, wherein The crystalline form is sodium deoxycholate crystalline form A, which shows characteristic peaks at 5.1, 10.2, 11.6, 12.7, 13.6, 15.5, 18.6, 20.1 and 22.0 in its X-ray powder diffraction spectrum expressed at a diffraction angle of 2θ±0.2°; optionally, it shows characteristic peaks at 5.1, 10.2, 11.6, 12.7, 13.6, 15.5, 17.9, 18.6, 19.4, 20.1, 22.0 and 23.8; optionally, it has an X-ray powder diffraction pattern substantially the same as that in FIG. 1; Optionally, the sodium deoxycholate form A has a water content of 0.1% w / w to 5% w / w.

3. The crystal form according to claim 1, wherein The crystalline form is sodium deoxycholate crystalline form B, which has characteristic peaks at 7.1, 7.8, 9.4, 11.7, 13.0, 14.4, 15.6, 17.9, 21.1 and 23.2 in its X-ray powder diffraction spectrum expressed at a diffraction angle of 2θ±0.2°; optionally, characteristic peaks are shown at 7.1, 7.8, 9.4, 11.7, 13.0, 14.4, 15.6, 17.9, 19.0, 21.1, 21.6, 23.2 and 23.7; optionally, it has an X-ray powder diffraction pattern substantially the same as that in FIG. 3; Optionally, the sodium deoxycholate form B has a water content of 0.1% w / w to 3% w / w.

4. The method for preparing the crystal form according to claim 1 or 2, wherein: The preparation method comprises the following steps: Mix deoxycholic acid and the first solvent, add sodium hydroxide, heat to 50°C-65°C to dissolve, cool to 40°C-55°C, filter to remove mechanical impurities; heat to 55°C-65°C, add the second solvent, cool to 10°C-30°C to crystallize for 1-3 hours, filter, and dry to obtain Form A.

5. The preparation method according to claim 4, wherein The first solvent is selected from at least one of water, acetone, tetrahydrofuran, 1,4-dioxane, methanol, ethanol or a combination thereof; optionally, the first solvent is water, a mixed solution of water and acetone, or a mixed solution of water and tetrahydrofuran.

6. The preparation method according to claim 4, wherein The second solvent is selected from at least one of water, acetone, tetrahydrofuran, 1,4-dioxane, methanol, ethanol or a combination thereof; optionally, the second solvent is acetone or tetrahydrofuran.

7. The preparation method according to claim 4, wherein The volume ratio of the second solvent to the first solvent is (1-50):(50-1), optionally, 1:3, 1:2, 1:1, 7:6, 2:1, 5:1, 10:1, 15:1, 20:1, 25:1, 30:1, 35:1, 40:1, 45:1, or 50:

1.

8. The method for preparing the crystal form according to claim 1 or 3, wherein: The preparation method comprises the following steps: The sodium deoxycholate crystal form A is mixed with the third solvent, and the obtained solution is concentrated under reduced pressure to obtain a solid, which is then dried to obtain the crystal form B.

9. The preparation method according to claim 8, wherein: The third solvent is selected from at least one of anhydrous methanol, anhydrous ethanol, isopropanol, n-propanol, n-butanol or a combination thereof; optionally, the third solvent is anhydrous methanol, anhydrous ethanol or isopropanol.

10. Use of the crystal form according to any one of claims 1 to 3 as a detergent, emulsifier or pharmaceutical excipient.

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