Preparation method of sugammadex double-oxidation impurity

Abstract

The invention relates to a preparation method and application of sugammadex double-oxide impurities, and the method comprises the following steps: under the conditions of illumination and an oxidizing agent, sugammadex is subjected to degradation reaction in a solvent, and chromatographic column separation is adopted to prepare the sugammadex double-oxide impurities. The preparation method disclosed by the invention is simple and easy to operate, mild in condition and high in reaction product purity, and an impurity reference substance meeting requirements can be provided for quality control of sugammadex, so that the safety and effectiveness of a sugammadex preparation are ensured.

Description

Technical Field

[0001] The invention belongs to the field of medicine, and in particular relates to a method for preparing sugammadex double-oxidized impurities. Background Art

[0002] The main active ingredient of Sugammadex Sodium Injection is a γ-cyclodextrin derivative, chemical name: 6 A ,6 B ,6 C ,6 D ,6 E ,7 F ,6 G ,6 H -Octane-S-(2-carboxyethyl)-6 A ,6 B ,6 C ,6 D ,6 E ,7 F ,6 G ,6 H -Octathio-γ-cyclodextrin octasodium salt, with the structural formula as Formula II, can form a water-soluble complex with the neuromuscular blocking drug rocuronium bromide at a ratio of 1:1, thereby blocking the effect of muscle relaxants, and has no adverse reactions related to anticholinesterase drugs. As the world's first selective muscle relaxant antagonist for reversing neuromuscular blockers, it can help patients with general anesthesia accurately and quickly reverse deep and moderate muscle relaxation in clinical surgical anesthesia, promote patients to restore their spontaneous breathing and limb activity ability, and help improve patients' postoperative outcomes. On July 25, 2008, the European Medicines Agency approved the marketing of Sugammadex Sodium Injection in Europe, and on October 20, 2010, Sugammadex Sodium Injection was approved for marketing in Japan. In November 2015, the FDA approved the drug for marketing in the United States. At present, on April 26, 2017, CFDA has approved the marketing of Sugammadex Sodium Injection in China, and the drug has a good market prospect.

[0003]

[0004] Since sugammadex sodium has a ring structure, there are many active sites in the side chains formed by the eight substitution reactions, which are very susceptible to oxidative degradation, posing potential risks to clinical injection administration.

[0005] Patent CN110105469A reports a method for preparing a monooxide impurity of sugammadex sodium, and a dioxide impurity, whose structural formula is as Formula I, is a potential degradation product of sugammadex sodium injection, and its research and content control are particularly important. At present, there is no literature reporting the preparation of dioxide impurities of sugammadex sodium injection. Therefore, it is urgent to develop a method for preparing disulfoxide impurities of sugammadex sodium injection to provide an impurity reference substance that meets the requirements for drug quality control. Summary of the invention

[0006] The technical problem solved by the present invention is to provide a method for preparing sugammadex double oxidized impurities, and to provide an impurity reference substance that meets the requirements for the quality control of sugammadex. The method is simple and easy to operate, has mild conditions, and has a high purity of the reaction product.

[0007] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions.

[0008] The invention discloses a method for preparing a sugammadex double-oxidized impurity, which is characterized in that: under the conditions of light and an oxidant, sugammadex undergoes a degradation reaction in a solvent, and the compound of formula I is obtained after separation and purification by preparative chromatography.

[0009] In the above-mentioned preparation method of the present invention, the solvent is water, methanol, acetonitrile, ethanol or a mixture thereof in any proportion, and the preferred solvent is water.

[0010] In the above-mentioned preparation method of the present invention, the lighting condition is natural light, ultraviolet light or fluorescent light, preferably fluorescent light, and more preferably a light intensity of 4500 Lux.

[0011] In the above-mentioned preparation method of the present invention, the illumination time is 2h to 5 days, preferably 2h.

[0012] In the above preparation method of the present invention, the oxidant is selected from oxygen, a gas mixture containing oxygen or hydrogen peroxide.

[0013] In the above-mentioned preparation method of the present invention, the degradation reaction solution is diluted with an acetonitrile aqueous solution before preparative chromatography separation and purification, preferably a 50% acetonitrile solution, and the pH value is adjusted to 3-5 with phosphoric acid, and the pH value is preferably 4.

[0014] In the preparation method of the present invention, the preparative chromatography, the preparative column is a C18 chromatographic column, the mobile phase is composed of 0.1~1% phosphoric acid solution-organic phase, wherein the volume ratio of phosphoric acid solution to organic phase is 75~85:25~15, the organic phase is selected from methanol, acetonitrile or a mixture of the two, preferably acetonitrile; the mobile phase flow rate is 5~40ml / min, preferably 20ml / min; the detection wavelength is 200~230nm, preferably 200m, and the column temperature is 15~65°C, preferably 30°C; The preparation method of the present invention further comprises collecting the target fraction of the preparative chromatography, blowing off the organic phase under nitrogen, desalting with a preparative column, collecting the fraction washed with 50% acetonitrile solution, blowing off the organic phase under nitrogen, and freeze-drying with a freeze dryer to obtain the sugammadex dioxidation impurity compound.

[0015] After a series of experimental studies, the inventors have developed the preparation method provided by the present invention, which has the following advantages: (1) Simple operation and high reaction efficiency. Sugammadex dioxidized impurities can be obtained by simply irradiating the sample solution with light and oxidative degradation.

[0016] (2) Easy purification and high purity. After the degradation solution is prepared by chromatography, freeze-dried and other operations, the purity of the obtained impurities can reach 95%. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 HPLC chromatogram of impurity purity of sugammadex double oxide; Figure 2 Mass spectrum of sugammadex dioxidized impurity; Figure 3 Sugammadex Dioxide Impurity 1 HNMR spectra; Figure 4 Sugammadex Dioxide Impurity 13 CNMR image. DETAILED DESCRIPTION

[0018] The present invention is further described in conjunction with the following examples to enable those skilled in the art to better understand the present invention, but the scope of the present invention is not limited in any way. Any equivalent changes or modifications made according to the spirit of the invention should be included in the protection scope of the present invention.

[0019] Example 1 Take an appropriate amount of 100 mg / ml sodium sugammadex solution, place it in an open container, and place it under a light intensity of 4500 Lux for 5 days. Dilute the resulting degradation solution with acetonitrile aqueous solution (adjust the pH value to 4 with phosphoric acid), and filter. Take the filtrate and continuously inject it under the following chromatographic conditions for preparative separation.

[0020] Chromatographic conditions: Collect all target fractions, blow off the organic phase under nitrogen, desalt with a preparative column, collect the fraction washed with 50% acetonitrile solution, blow off the organic phase under nitrogen. Prefreeze with a freeze dryer at -40°C, dry once at -10°C, and dry twice at 10°C to obtain the compound shown in Formula I with a chromatographic purity of 95.6%. The purity chromatogram is shown in Figure 1 ; Mass spectrum see Figure 2 , mass-to-charge ratio is 1015.1923, molecular formula is C 72 H112 O 50 S 8 .

[0021] Example 2 Take an appropriate amount of 10 mg / ml sodium sugammadex solution, add an appropriate amount of hydrogen peroxide, and place it under 4500 Lux light intensity for 2 hours. Dilute the resulting degradation solution with acetonitrile aqueous solution (adjust the pH value to 5 with phosphoric acid) and filter. Take the filtrate and continuously inject it under the following chromatographic conditions for preparative separation.

[0022] Chromatographic conditions: Collect all target fractions, blow off the organic phase under nitrogen, desalt with a preparative column, collect the fraction washed with 50% acetonitrile solution, blow off the organic phase under nitrogen. Prefreeze with a freeze dryer at -40°C, dry once at -10°C, and dry twice at 10°C to obtain the compound shown in Formula I with a chromatographic purity of 93.9%. 1 H-NMR Figure 3 , 13 C-NMR Figure 4 .

[0023] Example 3 Take an appropriate amount of 50 mg / ml sodium sugammadex solution and place it in an open container at 69.5 μW / cm 2 Place under UV conditions for 2 days. Dilute the resulting degradation solution with acetonitrile aqueous solution (adjust pH to 3 with phosphoric acid), filter. Take the filtrate and continuously inject it under the following chromatographic conditions for preparative separation.

[0024] Chromatographic conditions: Collect all target fractions, blow off the organic phase under nitrogen, desalt with a preparative column, collect the fraction washed with 50% acetonitrile solution, blow off the organic phase under nitrogen, pre-freeze with a freeze dryer at -40°C, dry once at -10°C, and dry twice at 10°C to obtain the compound shown in Formula I with a chromatographic purity of 95.1%.

Claims

1. A method for preparing a sugammadex dioxidized impurity compound of formula I, characterized in that The method comprises: under the conditions of light and an oxidant, sugammadex undergoes a degradation reaction in a solvent, and the compound of formula I is obtained after separation and purification by preparative chromatography, 。 2. The preparation method according to claim 1, wherein the solvent is water, methanol, acetonitrile, ethanol or a mixture thereof in any proportion, preferably water.

3. The preparation method according to claim 1, wherein the illumination condition is natural illumination, ultraviolet light illumination or fluorescent light illumination, preferably fluorescent light illumination, more preferably a light intensity of 4500 Lux, and the illumination time is 2 h to 5 days, preferably 2 h.

4. The preparation method according to claim 1, wherein the oxidant is selected from oxygen, a gas mixture containing oxygen or hydrogen peroxide.

5. The preparation method according to claim 1, wherein the degradation reaction solution is diluted with an acetonitrile aqueous solution before the preparative chromatography separation and purification, and the pH value is adjusted to 3-5 with phosphoric acid, and the pH value is preferably 4.

6. The preparation method according to claim 5, wherein the preparative chromatography column is a C18 chromatographic column, the mobile phase consists of 0.1-1% phosphoric acid solution-organic phase, the mobile phase flow rate is 5-40 ml / min, the detection wavelength is 200-230 nm, and the column temperature is 15-65°C.

7. The preparation method according to claim 6, wherein the mobile phase flow rate is 20 ml / min, the detection wavelength is 200 nm, and the column temperature is 30°C.

8. The preparation method according to claim 6, wherein the volume ratio of the phosphoric acid solution to the organic phase is 75-85:25-15, preferably 80:

20.

9. The preparation method according to claim 8, wherein the organic phase is selected from methanol, acetonitrile or a mixture of the two, preferably acetonitrile.

10. The preparation method according to claims 1 to 9, further comprising collecting the target fraction of the preparative chromatography, blowing off the organic phase under nitrogen, desalting with a preparative column, collecting the fraction washed with 50% acetonitrile solution, blowing off the organic phase under nitrogen, and freeze-drying with a freeze dryer to obtain the sugammadex dioxidation impurity compound.

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