Method for detecting related substances of crisaborole raw material medicine

Abstract

The invention relates to a method for detecting related substances in a crisaborole bulk drug, and discloses a method for detecting the crisaborole bulk drug and impurities thereof by adopting a high performance liquid chromatography, the method adopts a chromatographic column as a silica gel bonded octadecylsilane column, and adopts a certain amount of acidic reagent solution-organic phase as a mobile phase; and carrying out separation and detection by adopting an isocratic elution mode. According to the method for detecting the related substances of the crisaborole bulk drug, five impurities of the bulk drug can be detected at a time, the specificity is high, the precision is high, the repeatability is good, and the quality of the crisaborole bulk drug and the impurities of the crisaborole bulk drug can be effectively controlled.

Description

Technical Field

[0001] The present invention relates to the technical field of chemical drug analysis, and in particular to an analytical method for analyzing and determining crisaborole raw material drugs and related impurities by liquid chromatography. Background Art

[0002] Crisaborole, whose English name is Crisaborole, has a chemical name of 4-[(1,3-dihydro-1-hydroxy-2,1-benzoxaborol-5-yl)oxy]benzonitrile. Its English name is 4-[(1,3-dihydro-1-hydroxy-2,1-benzoxaborol-5-yl)oxy]benzonitrile. Its molecular formula is C14H10BNO3 and its molecular weight is 251.05.

[0003] Its structural formula is as follows:

[0004] Criborole was first developed by Anacor Pharmaceuticals. Pfizer acquired Anacor in May 2016. In December of the same year, the US FDA approved Criborole ointment (2% or 5%) for the topical treatment of mild to moderate atopic dermatitis in patients over 2 years old. The trade name is Eucrisa.

[0005] While crisaborole impurities have similar structures, their polarities vary. This makes separation of crisaborole and the intermediates introduced during crisaborole synthesis extremely challenging. If these intermediates are not completely removed, they can cause side reactions that can affect the purity and quality of the drug. Therefore, separation of crisaborole and other related substances is crucial for quality control during crisaborole synthesis. Summary of the Invention

[0006] The present invention proposes a method for analyzing related substances of crisaborole raw material medicine, thereby being able to quickly, effectively and accurately monitor the purity of related substances of crisaborole raw material medicine, solving the problem of separation and detection of crisaborole and its impurities, thereby achieving quality control of its final product raw material medicine.

[0007] The present invention provides a method for analyzing related substances of a crisaborole raw material drug, which adopts high performance liquid chromatography, uses a silica gel-bonded octadecylsilane column as the chromatographic column, uses a certain amount of acidic reagent solution as mobile phase A and an organic phase as mobile phase B, and performs isocratic elution.

[0008] The present invention provides a method for analyzing related substances of crisaborole raw materials, wherein the isocratic elution procedure is:

[0009] Preferably, the certain amount of acidic reagent solution is 0.1% trifluoroacetic acid solution.

[0010] Preferably, the organic phase is acetonitrile.

[0011] Preferably, the chromatographic column provided is ShimNex HE C18 (4.6 mm×150 mm, 5 μm).

[0012] The separation and determination method of the present invention can be achieved by the following method: 1) Take an appropriate amount of crisaborole sample and dissolve it in a diluent to prepare a sample solution containing 0.2 mg of crisaborole per 1 ml; 2) Set the mobile phase flow rate to 0.5-1.5 mL / min, preferably 1.0 mL / min, the detection wavelength to 240-260 nm, with the optimal detection wavelength being 254 nm, and the column oven temperature to 20-40°C, with the optimal column oven temperature being 30°C. 3) Take 5-20 μl of the sample solution from 1) above and inject it into a liquid chromatograph to separate and determine crisaborole and related substances. High-performance liquid chromatograph: Agilent 1260 InfinityII; Column: ShimNex HE C18 (4.6 mm × 150 mm, 5 µm); Mobile phase A: 0.1% trifluoroacetic acid solution (1 ml of trifluoroacetic acid was added to 1000 ml of water, mixed, filtered, and ultrasonically degassed to obtain the result); Mobile phase B: acetonitrile; Elution was performed according to the following gradient:

[0013] Flow rate: 1.0 mL / min; Column temperature: 30°C; Detection wavelength: 254nm; Injection volume: 10 μl.

[0014] The invention uses a ShimNex HE C18 (150×4.6 mm, 5μm) chromatographic column to effectively separate crisaborole and its related substances. This solves the problem of separating and determining crisaborole and its related impurities, ensuring the quality control of crisaborole. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] Figure 1 This is the HPLC chart of the blank solvent of crisaborole in Example 1.

[0016] Figure 2This is the HPLC chart of crisaborole and its related impurities in Example 1.

[0017] Figure 3 This is the HPLC chart of the reproducibility of crisaborole in Example 1. DETAILED DESCRIPTION The following examples are provided to further understand the present invention, but are not intended to limit the scope of this invention.

[0019] Example 1 Instruments and conditions High-performance liquid chromatograph: Agilent 1260 InfinityII; Column: ShimNex HE C18 (4.6 mm × 150 mm, 5 µm); Mobile phase A: 0.1% trifluoroacetic acid solution (1 ml of trifluoroacetic acid was added to 1000 ml of water, mixed, filtered, and ultrasonically degassed to obtain the result); Mobile phase B: acetonitrile; Elution was performed according to the following gradient:

[0020] Flow rate: 1.0 mL / min; Column temperature: 30°C; Detection wavelength: 254nm; Injection volume: 10 μl; The HPLC detection steps of related substances of crisaborole raw materials are as follows: Take an appropriate amount of crisaborole and its impurity reference substances, dissolve the sample with a diluent, and prepare a sample solution containing 0.2 mg / mL of crisaborole and about 2 µg / mL of its impurities.

[0021] Take 10µl of the above solution and inject it into the liquid chromatograph to record the chromatogram.

[0022] Specificity test results:

[0023] Repeatability test results:

[0024] In summary, the present invention can separate crisaborole from its impurities and can accurately perform quantitative detection, thereby effectively controlling the product quality of crisaborole.

Claims

1. A method for detecting related substances of crisaborole raw materials, using high performance liquid chromatography, the chromatographic column is a silica gel bonded octadecylsilane column, a certain amount of acidic reagent solution-organic phase is used as the mobile phase, and gradient elution is performed.

2. According to the method for analyzing and detecting related substances of crisaborole raw material drug according to claim 1, the organic phase is selected from one of the following compounds: methanol and acetonitrile.

3. According to the method for analyzing and detecting related substances in the crisaborole raw material drug of claim 1, a certain amount of acidic reagent solution is selected from one of the following inorganic salts: perchloric acid, trifluoroacetic acid.

4. According to the method for analyzing and detecting related substances of the crisaborole raw material drug of claim 1, the isocratic process is a gradient elution as follows:

5. According to the method for analyzing and detecting related substances of crisaborole raw materials according to claim 1, the chromatographic column preferably has a length of 150 mm, a diameter of 4.6 mm, and a filler particle size of 5 µm.

6. According to the method for analyzing and detecting related substances of crisaborole raw material drug according to claims 1 and 3, the certain amount of acidic reagent solution is preferably 0.1% trifluoroacetic acid solution.

7. According to the method for analyzing and detecting related substances of the crisaborole raw material drug according to claims 2 and 4, the organic phase is preferably acetonitrile.

8. The method for analyzing and detecting related substances of the crisaborole raw material drug according to claim 1, comprising the following steps: ① setting the flow rate to 0.5-1.5 ml / min; ② setting the detection wavelength to 240-260 nm; ③ setting the column temperature to 20-40°C; ④ setting the injection volume to 5-20 µl.

9. According to the method for analyzing and detecting related substances of crisaborole raw materials as described in claim 8, the chromatographic conditions are as follows: ① the flow rate is preferably 1.0 ml / min; ② the wavelength is preferably 254 nm; ③ the column temperature is preferably 30°C; ④ the injection volume is preferably 10 µl.

10. According to the method for analyzing related substances of crisaborole raw material medicine in claim 1, the related substance impurities are:

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