Micafungin sodium medicine impurity as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of biological medicine, particularly relates to a novel micafungin sodium drug impurity compound, and further discloses a preparation method and application thereof. The micafungin sodium drug impurity has a structure as shown in a formula (I), and the purity of the micafungin sodium drug impurity is greater than 95%. The high-purity impurity can serve as a reference substance for establishing an analysis method to be used for qualitative and quantitative analysis of impurities in micafungin sodium medicine, and technical support is provided for quality research of micafungin sodium.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, specifically relating to a new micafungin sodium drug impurity compound, and further disclosing its preparation method and application. Background Technology

[0002] In recent years, with the increasing incidence of cancer chemotherapy, organ transplantation, and AIDS, the incidence of fungal infections has risen significantly, especially the incidence of deep fungal infections and the resulting mortality rate, which continues to rise, seriously threatening human health and becoming one of the most challenging clinical problems. Echinocandins B are novel drugs developed in the early 21st century, possessing a completely new mechanism of action, a broad antifungal spectrum, no cross-resistance, and no significant adverse reactions caused by their mechanism of action. They are among the safest antifungal drugs to date and are currently considered ideal for treating deep fungal infections.

[0003] Micafungin sodium is a new generation of echinocandin antifungal drug, primarily used to treat fungemia, respiratory fungal infections, and gastrointestinal fungal infections caused by Aspergillus and Candida. It is also used to prevent Aspergillus and Candida infections in hematopoietic stem cell patients. As a type of echinocandin antifungal drug, it inhibits the synthesis of β(1,3)-glucan in filamentous fungi and yeasts. In recent years, the incidence and mortality rates of fungal infections have shown a year-on-year upward trend, posing a fatal threat, especially to critically ill patients. Micafungin sodium, as a echinocandin, has good antifungal activity and is the first-line drug for treating infections caused by Candida or Aspergillus. It has good therapeutic effects, minimal impact on human cells, and exhibits low toxicity and high efficacy in clinical practice.

[0004] In existing technologies, micafungin sodium is a semi-synthetic product, obtained by fermentation to obtain the parent nucleus FR179642, which is then prepared by chemical synthesis through a condensation reaction with the active side chain. Because FR179642 has a polypeptide parent ring structure containing multiple amino and hydroxyl functional groups, various byproducts are generated during the reaction with the active side chain. Ohigashi et al. (Journal of Synthetic Organic Chemistry, vol 64, No. 12, 2006) reported in 2006 that due to the large number of active groups in the micafungin parent nucleus, a side reaction of monoacylation could be detected during the acylation reaction to prepare echinocandin, potentially indicating the presence of diacylated impurities. However, the structures of possible diacylated impurities were not reported, nor were related preparation and purification methods reported. The parent nucleus FR179642 (Formula II) contains 10 reactive sites (hydroxyl and amino groups), resulting in 90 possible diacylated impurities. CN202411642606.5 discloses a diacylated impurity of micafungin sodium, having the structure shown in formula (Ⅳ), but does not disclose whether the numerous other hydroxyl functional groups on the parent core FR179642 structure can also generate diacylated impurities.

[0005]

[0006] Impurity analysis is crucial in drug development. Impurities are closely related to the quality, safety, and stability of drugs. Preparing and structurally confirming impurities helps us understand their formation pathways, providing a basis for improving drug synthesis routes and manufacturing processes. Impurities can also be used in drug quality control, serving as reference standards for establishing analytical methods. For pharmaceutical manufacturers, strict control over impurities is essential.

[0007] Therefore, research on impurities in micafungin sodium can be used for the qualitative and quantitative analysis of impurities in the production of micafungin sodium, thereby providing technical support for subsequent quality research on micafungin sodium.

[0008] Therefore, there is a need in the field to develop an efficient and convenient method for preparing high-purity micafungin sodium drug impurities, which would be of positive significance for the development of micafungin sodium. Summary of the Invention

[0009] The first technical problem this invention aims to solve is to provide a micafungin sodium drug impurity with an HPLC purity of over 95%, which can be used for the quality control of micafungin sodium and as a reference standard for establishing analytical methods. The second technical problem this invention aims to solve is to provide a method for preparing the aforementioned impurity.

[0010] To address the aforementioned technical problems, this invention discloses a micafungin sodium drug impurity compound or an acceptable salt thereof, wherein the micafungin sodium drug impurity compound has the structure shown in formula (I):

[0011]

[0012] The chemical name of the micafungin sodium drug impurity compound with the structure shown in formula (Ⅰ) provided by this invention is: 5-[(1S,2S)-2-[(3S,6S,9S,11R,15S,18S,20R,21R,24S,25S,26S)-3-[(R)-2-carbamoyl-1-hydroxyethyl]-11,20,21,25-tetrahydroxy-15-[(R)-1-hydroxyethyl]-26-methyl-2,5,8,14,17,23-hexaoxo-18-[4-[5-(4-pentoxyphenyl)isoxazolyl-3-yl]benzoylamino]-1,4,7,13,16,22-hexaazatricyclo[22.3.0.0] 9,13 Sodium hexahexa-6-yl]-1,2-dihydroxyethyl]-2-[4-[5-(4-pentoxyphenyl)isoxazolyl-3-yl]benzoyloxy]phenyl sulfate.

[0013] The present invention also discloses a high-purity micafungin sodium drug impurity, comprising the micafungin sodium drug impurity compound of formula (I) or an acceptable salt thereof, wherein the HPLC purity reaches 95% or higher.

[0014] This invention also discloses a method for preparing the high-purity micafungin sodium drug impurity, comprising the following steps:

[0015] (1) The compound FR179642, which has the following formula (Ⅱ), is added to an organic solvent and mixed.

[0016]

[0017] (2) Continue to add organic base and active ester of micafungin sodium side chain as shown in formula (III) to obtain micafungin sodium drug impurity with the structure shown in formula (I) after reaction;

[0018]

[0019] Specifically, in the method for preparing the high-purity micafungin sodium drug impurity, the organic solvent in step (1) includes at least one of N,N-dimethylformamide, N,N-dimethylacetamide, and dimethyl sulfoxide; the volume of the organic solvent used is 8 to 10 times the weight of compound FR179642, in ml / g.

[0020] Specifically, in the method for preparing the high-purity micafungin sodium drug impurity, the organic base in step (2) includes at least one of triethylamine, N,N-diisopropylethylamine, and 4-dimethylaminopyridine.

[0021] Specifically, in the method for preparing the high-purity micafungin sodium drug impurity, the amount of organic base added in step (2) is 1.6 to 2.0 equivalents of the molar amount of compound FR179642; preferably 1.6, 1.8 or 2.0 equivalents.

[0022] Specifically, in the method for preparing the high-purity micafungin sodium drug impurity, the amount of micafungin sodium side-chain active ester added in step (2) is 1.5 to 2.0 equivalents of the molar amount of compound FR179642; preferably 1.5, 1.7, 1.9 or 2.0 equivalents.

[0023] Specifically, in the method for preparing the high-purity micafungin sodium drug impurity, the reaction temperature in step (2) is 0-10℃.

[0024] In previous research on micafungin sodium, this invention detected diacylated impurities during the condensation reaction, but their low concentration made purification and enrichment difficult. With optimized reaction conditions, the proportion of this impurity compound in the HPLC chromatogram of the reaction solution continuously increased, providing favorable conditions for impurity purification and enrichment.

[0025] This invention is the first to prepare micafungin sodium drug impurities with a purity of 95-99% or higher using the structure of formula (Ⅰ), which can be applied to the quality control of micafungin sodium and can be used as a reference standard for analytical methods. Attached Figure Description

[0026] Figure 1 This is the HPLC chromatogram of the drug impurities in micafungin sodium in Example 4.

[0027] Figure 2 This is the 1H NMR spectrum of the drug impurities in micafungin sodium from Example 4. 1 H-NMR).

[0028] Figure 3 This is the carbon NMR spectrum of the drug impurities in micafungin sodium in Example 4. 13 C-NMR).

[0029] Figure 4 This is a high-resolution mass spectrum (HRMS) of the drug impurities in micafungin sodium in Example 4. Detailed Implementation

[0030] The following embodiments further illustrate the present invention. However, it should be noted that the following embodiments are only used to describe the content of the invention and do not constitute a limitation on the scope of protection of the present invention. The scope of protection of the present invention shall be determined by the claims.

[0031] In the following examples, unless otherwise specified, all temperatures are in Celsius; unless otherwise specified, all starting materials and reagents are commercially available and used directly without further purification; unless otherwise specified, all solvents are industrial grade solvents and used directly without further treatment.

[0032] The nuclear magnetic resonance spectrometer involved in this invention is the BRUKER 400MHz model.

[0033] The high-resolution mass spectrometer involved in this invention is an Aglient 1260LC-6540Q-TOF MS.

[0034] The HPLC detection instrument involved in this invention is the Aglient 1200 model.

[0035] The FR179642 and micafungin sodium side chain active ester used in this invention are both produced by Zhejiang Hisun Pharmaceutical Co., Ltd.

[0036] The HPLC method used in this invention for detecting purity is as follows:

[0037] Column: Agilent SB-C18, 4.6×150mm, 5μm or equivalent column.

[0038] Mobile phase: 0.1 mol / L potassium dihydrogen phosphate solution: acetonitrile = 50:50.

[0039] Flow rate: 2.0 ml / min.

[0040] Detection wavelength: 273nm.

[0041] Running time: Approximately 20 minutes.

[0042] The preparative column purification and separation method used in this invention is as follows:

[0043] Preparation column: DAC100.

[0044] Packing material: C18 silica gel packing material.

[0045] Mobile phase: Acetonitrile: 100 mmol sodium chloride aqueous solution = 75: 25.

[0046] Detection wavelength: 270nm.

[0047] Flow rate: 280 ml / min.

[0048] Example 1

[0049] Take 1.0 g of FR179642 and add it to a reaction flask containing 8 ml of N,N-dimethylformamide. Stir and cool to 0-10 °C, continue stirring until dissolved. Add 232 μl (1.6 equivalents) of triethylamine and stir for about 5 min. Then add 0.83 g (1.7 equivalents) of the active ester of micafungin sodium side chain and continue stirring at 0-10 °C for 10 h. HPLC detection showed that micafungin sodium drug impurity with the structure of formula (I) was generated.

[0050] The above reaction solution was filtered, and 48 ml of ethyl acetate was added dropwise to the filtrate under stirring. The mixture was stirred for 1 hour and then filtered again. The filter cake was dissolved in water and purified by preparative column chromatography. The collected solution was concentrated to remove acetonitrile, and impurities precipitated out. The solution was filtered and dried to obtain 151 mg of micafungin sodium drug impurity white powder with the structure of formula (I). The purity was determined by HPLC to be 97.28%.

[0051] Example 2

[0052] Take 1.0 g of FR179642 and add it to a reaction flask containing 10 ml of N,N-dimethylacetamide. Stir and cool to 0-10 °C, continue stirring until dissolved. Add 363 μl (2.0 equivalent) of N,N-diisopropylethylamine and stir for about 5 min. Then add 0.73 g (1.5 equivalent) of the active ester of micafungin sodium side chain and continue stirring at 0-10 °C for 8 h. HPLC detection showed that micafungin sodium drug impurity with the structure of formula (I) was generated.

[0053] The above reaction solution was filtered, and 60 ml of ethyl acetate was added dropwise to the filtrate under stirring. The mixture was stirred for 1 hour and then filtered again. The filter cake was dissolved in water and purified by preparative column chromatography. The collected solution was concentrated to remove acetonitrile, and impurities precipitated out. The solution was filtered and dried to obtain 138 mg of micafungin sodium drug impurity white powder with the structure of formula (I). The purity was 99.14% as determined by HPLC.

[0054] Example 3

[0055] Take 1.0 g of FR179642 and add it to a reaction flask containing 10 ml of dimethyl sulfoxide. Stir and cool to 0-10 °C, continue stirring until dissolved. Add 0.23 g (1.8 equivalents) of 4-dimethylaminopyridine and stir for about 5 min. Then add about 0.98 g (2.0 equivalents) of the active ester of micafungin sodium side chain and continue stirring at 0-10 °C for 8 h. HPLC detection showed that micafungin sodium drug impurities with the structure of formula (I) were generated.

[0056] The above reaction solution was filtered, and 60 ml of ethyl acetate was added dropwise to the filtrate under stirring. The mixture was stirred for 1 hour and then filtered again. The filter cake was dissolved in water and purified by preparative column chromatography. The collected solution was concentrated to remove acetonitrile, and impurities precipitated out. The solution was filtered and dried to obtain 218 mg of micafungin sodium drug impurity white powder with the structure of formula (I). The purity was determined by HPLC to be 98.31%.

[0057] Example 4

[0058] Take 5.0 g of FR179642 and add it to a reaction flask containing 50 ml of N,N-dimethylformamide. Stir and cool to 0-10 °C, continue stirring until dissolved. Add 1.3 mL (1.8 equivalents) of triethylamine and stir for about 5 min. Then add about 4.6 g (1.9 equivalents) of the active ester of micafungin sodium side chain and continue stirring at 0-10 °C for 10 h. HPLC detection showed that micafungin sodium drug impurities with the structure of formula (I) were generated.

[0059] The above reaction solution was filtered, and 300 ml of ethyl acetate was added dropwise to the filtrate under stirring. The mixture was stirred for 1 hour and then filtered again. The filter cake was dissolved in water and purified by preparative column chromatography. The collected liquid was concentrated to remove acetonitrile, and impurities precipitated out. After filtration and drying, 1.2 g of micafungin sodium drug impurity white powder with the structure of formula (I) was obtained. The purity was determined by HPLC to be 99.55% (see Appendix). Figure 1 ).

[0060] The micafungin sodium drug impurity obtained in this embodiment, with the structure of formula (Ⅰ), was confirmed by nuclear magnetic resonance spectroscopy and mass spectrometry, and the data are as follows:

[0061] 1H-NMR (DMSO-d6): 0.91 (t, 6H), 0.98 (d, 3H), 1.10 (d, 3H), 1.39 (m, 8H), 1.75 (m, 4H), 1.95 (m, 3H), 2.28 (m, 2H), 2.38 (m, 1H), 2.57 (m, 1H), 3.22 (t, 1H), 3.79 (m, 2H), 3.95 (m, 1H), 4.06 (m, 7H), 4.26 (m, 2H), 4.32 (s, 1H), 4.40 (m, 4H), 4.47 (m, 1H), 4.84 (m, 2H), 4.94 (m, 1H), 5.13 (m, 5H), 5.19 (d, 1H), 5.46 (d, 1H), 5.56 (d, 1H), 6.90 (s, 1H), 7.03 (d, 1H), 7.12 (dd, 4H), 7.20 (d, 1H), 7.30 (s, 1H), 7.37 (d, 1H), 7.48 (d, 1H), 7.56 (s, 1H), 7.58 (s, 1H), 7.61 (s, 1H), 7.8​​​​​​​​​​​​The peak (see appendix) Figure 4 ).

Claims

1. A compound that is a micafungin sodium pharmaceutical impurity, or an acceptable salt thereof, characterized in that, The micafungin sodium drug impurity has the following formula (I) structure:

2. A high purity micafungin sodium drug impurity characterized in that, The micafungin sodium drug impurity compound or its acceptable salt as claimed in claim 1 has an HPLC purity of more than 95%.

3. A process for the preparation of a micafungin sodium pharmaceutical impurity as claimed in claim 1 or 2, characterized by, The method comprises the following steps: (1) adding a compound FR179642 having the following formula (II) into an organic solvent mixture; (2) continuously adding an organic base and a micafungin sodium side chain active ester having the following formula (III) to obtain the micafungin sodium drug impurity having the structure of formula (I) as claimed in claim 1 through reaction; 4. The production method according to claim 3, characterized by, The organic solvent in the step (1) comprises at least one of N,N-dimethylformamide, N,N-dimethylacetamide and dimethyl sulfoxide; the volume of the organic solvent is 8-10 times the weight of the compound FR179642, in units of ml / g.

5. The process for the preparation of high purity micafungin sodium drug impurities according to claim 3 or 4, characterized in that, The organic base in the step (2) comprises at least one of triethylamine, N,N-diisopropyl ethylamine and 4-dimethylamino pyridine.

6. The method of any one of claims 3-5, wherein, The addition amount of the organic base in the step (2) is 1.6-2.0 equivalents of the molar amount of the compound FR179642; preferably 1.6, 1.8 or 2.0 equivalents.

7. The method of any one of claims 3-6, wherein, The addition amount of the micafungin sodium side chain active ester in the step (2) is 1.5-2.0 equivalents of the molar amount of the compound FR179642; preferably 1.5, 1.7, 1.9 or 2.0 equivalents.

8. The process for the preparation of high purity micafungin sodium drug impurities according to any one of claims 3 to 7, characterized in that, The reaction temperature in the step (2) is 0-10℃.

9. Use of the micafungin sodium pharmaceutical impurity of the structure of formula (I) according to claim 1 in pharmaceutical quality control, characterized in that The micafungin sodium drug impurity having the structure of formula (I) is used as a control for establishing an analysis method.

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