阅读说明：本技术 与洛铂有关物质的检测方法 (Method for detecting lobaplatin-related substance ) 是由 窦啟玲 汪立冬 常新亮 于 2019-03-19 设计创作，主要内容包括：本发明涉及与铂类有关物质的检测方法。本发明提供了一种与洛铂有关物质的检测方法,其中,所述与洛铂有关物质包括式Ⅰ和Ⅱ结构的化合物,所述式Ⅰ结构为<Image he="225" wi="671" file="DDA0001999876170000011.GIF" imgContent="drawing" imgFormat="GIF" orientation="portrait" inline="no"></Image>所述式Ⅱ结构为<Image he="213" wi="562" file="DDA0001999876170000012.GIF" imgContent="drawing" imgFormat="GIF" orientation="portrait" inline="no"></Image>所述的检测方法为HPLC-MS法或者HPLC法,所述HPLC-MS法的检测条件为：用十八烷基硅烷键合硅胶为填充剂,以10-12mmol/L的乙酸铵为流动相A,甲醇：乙腈的体积比例＝1:(0.8-1.2)为流动相B,进行梯度洗脱。本发明建立了具有式Ⅰ和Ⅱ结构的铂类化合物作为洛铂质量标准中的有关物质进行检测的方法,以建立洛铂质量检测体系,该方法灵敏度高,专属性强,重复性好。(The invention relates to a method for detecting platinum-related substances. The invention provides a method for detecting a lobaplatin related substance, wherein the lobaplatin related substance comprises compounds with structures shown in formulas I and II, and the structure of the formula I is shown in the specification The structure of the formula II is The detection method is an HPLC-MS method or an HPLC method, and the detection conditions of the HPLC-MS method are as follows: octadecylsilane chemically bonded silica is used as a filling agent, 10-12mmol/L ammonium acetate is used as a mobile phase A, and methanol: acetonitrile with volume ratio of 1 (0.8-1.2) as mobile phase B, and gradient elution is carried out. The invention establishes platinum compounds with structures shown as formulas I and II as related substances in lobaplatin quality standard for detectionThe method is used for establishing a lobaplatin quality detection system, and has the advantages of high sensitivity, strong specificity and good repeatability.)

1. A method for detecting a lobaplatin-related substance, wherein the lobaplatin-related substance comprises compounds with structures shown in formulas I and II, and the structure of the formula I is shown in the specification

The structure of the formula II is

2. The detection method according to claim 1, wherein the detection method is an HPLC-MS method or an HPLC method.

3. The detection method according to claim 2, wherein the detection conditions of the HPLC-MS method are as follows: octadecylsilane chemically bonded silica is used as a filling agent, 10-12mmol/L ammonium acetate is used as a mobile phase A, and methanol: the volume ratio of acetonitrile is 1, (0.8-1.2) is used as a mobile phase B, and gradient elution is carried out; preferably, the mobile phase A is a 10mmol/L ammonium acetate solution, and the mobile phase B is methanol: the volume ratio of acetonitrile is 1:1.

4. The detection method according to claim 3, wherein the gradient elution pattern in the HPLC-MS method is as follows:

0-10 minutes: mobile phase a decreased from 95% to 40% by volume and mobile phase B increased from 5% to 60% by volume;

10-15 minutes: mobile phase a decreased from 40% to 10% by volume and mobile phase B increased from 60% to 90% by volume;

15-16 minutes: mobile phase a increased from 10 vol% to 95 vol%, and mobile phase B decreased from 90 vol% to 5 vol%;

16-24 minutes: 95 vol% mobile phase a: 5 vol% mobile phase B;

wherein, each time range of the gradient elution can be increased by 1-2 minutes or the time range of the gradient elution from 10-15 minutes can be decreased by 1-2 minutes.

5. The detection method according to any one of claims 2 to 4, wherein the MS condition in HPLC-MS is the use of an electrospray ion source.

6. The test method according to any one of claims 2 to 5, wherein the flow rate is 0.8-1.2ml, preferably 1.0ml per minute; preferably, the column temperature is 38-42 deg.C, preferably 40 deg.C.

7. The detection method according to any one of claims 1 to 6, wherein in a chromatogram of a system suitability test solution, a separation degree of a peak of the lobaplatin-related substance from its neighboring related substance is not less than 1.5; preferably, the relative standard deviation of the peak areas of the compound with the structure of the formula I and the compound with the structure of the formula II is not more than 10.0 percent.

8. The detection method according to any one of claims 1 to 7, wherein the mass spectrum of the test solution contains the compounds having the structures of formula I and formula II, and the peak areas of the compounds are not larger than the peak areas of the compounds having the structures of formula I and formula II in the control solution.

9. The detection method according to any one of claims 1 to 8, wherein the lobaplatin comprises either one or both of lobaplatin diastereomer I and lobaplatin diastereomer II.

Technical Field

The invention relates to the field of medicines, in particular to a method for detecting substances related to lobaplatin, belonging to the technical field of medicine analysis quality control.

Background

Lobaplatin (Lobaplatin, D19466), also known as Lobaplatin, is a third-generation platinum-based antitumor drug following cisplatin and carboplatin, and its chemical name is: cis- [ trans-1, 2-cyclobutanebis (methylamine) -N, N']- [ (2S) -lactic acid-O1, O2]-platinum (II), formula C₉H₁₈N₂O₃Pt has a molecular weight of 397.34 and a chemical structural formula shown in the following formula (a):

lobaplatin has alkylating effect, belongs to alkylating agent (broad sense), and has good antitumor effect, such as inhibiting in vitro AH 135-tumor, B16-melanoma, colon cancer 115, and in vivo mouse P338 leukemia. Lobaplatin is characterized by strong anticancer activity, low toxicity, no accumulative toxicity and renal toxicity and less toxicity to bone marrow, and currently marketed lobaplatin for injection is mainly used for treating breast cancer, small cell lung cancer and chronic myelogenous leukemia.

Disclosure of Invention

In order to ensure the safety, effectiveness and controllable quality of the medicine, the research on related substances and detection methods of the related substances is very important. Aiming at the drug, due to the existence of three chiral carbons and related substances generated in the preparation process, finding a suitable detection method for controlling the product quality of the drug is a technical problem which is urgently needed to be solved in the field.

The technical problem to be solved by the invention is to provide a new detection method to establish the detection of related substances in the lobaplatin so as to carry out quality control on the lobaplatin compound.

One skilled in the art will recognize that any substance that affects the purity of a drug is collectively referred to as a related substance. Research on related substances is an important part of drug development, and comprises selecting a proper analysis method, accurately distinguishing and determining the content of the related substances, and determining the reasonable limit of the related substances by combining the results of pharmaceutical, toxicological and clinical researches. This study is throughout the entire process of drug development.

Specifically, the present invention is realized by the following technical means.

The invention provides a method for detecting a lobaplatin related substance, wherein the lobaplatin related substance comprises compounds with structures shown in formula I and formula II, and the structure of the formula I is shown in the specification

The structure of the formula II is

Preferably, the detection method is an HPLC-MS method or an HPLC method.

Preferably, in the detection method, the detection conditions of the HPLC-MS method are: octadecylsilane chemically bonded silica is used as a filling agent, 10-12mmol/L ammonium acetate is used as a mobile phase A, and methanol: the volume ratio of acetonitrile is 1, (0.8-1.2) is used as a mobile phase B, and gradient elution is carried out; preferably, the mobile phase A is a 10mmol/L ammonium acetate solution, and the mobile phase B is methanol: the volume ratio of acetonitrile is 1:1.

Preferably, in the detection method, the gradient elution pattern in the HPLC-MS method is as follows:

0-10 minutes: mobile phase a decreased from 95% to 40% by volume and mobile phase B increased from 5% to 60% by volume;

10-15 minutes: mobile phase a decreased from 40% to 10% by volume and mobile phase B increased from 60% to 90% by volume;

15-16 minutes: mobile phase a increased from 10 vol% to 95 vol%, and mobile phase B decreased from 90 vol% to 5 vol%;

16-24 minutes: 95 vol% mobile phase a: 5 vol% mobile phase B;

wherein, each time range of the gradient elution can be increased by 1-2 minutes or the time range of the gradient elution from 10-15 minutes can be decreased by 1-2 minutes;

for example, the time range corresponding to gradient elution may be 0 to 11 minutes (or 0 to 12 minutes), 11 to 16 minutes (or 12 to 17 minutes), 16 to 17 minutes (or 17 to 18 minutes), 17 to 25 minutes (or 18 to 26 minutes); the time may be 0 to 10 minutes, 9 to 14 minutes (or 8 to 13 minutes), 14 to 15 minutes (or 13 to 14 minutes), or 15 to 23 minutes (or 14 to 22 minutes).

Preferably, in the detection method, an electrospray ion source is used as the MS condition in the HPLC-MS.

Preferably, for the detection method described above, wherein the flow rate is 0.8-1.2ml per minute, preferably 1.0 ml; preferably, the column temperature is 38-42 deg.C, preferably 40 deg.C.

Preferably, in the detection method described above, in which, in a chromatogram of the system suitability test solution, a degree of separation of a peak of the related substance from a peak of the related substance adjacent thereto is not less than 1.5; preferably, the relative standard deviation of the peak areas of the compound with the structure of the formula I and the compound with the structure of the formula II is not more than 10.0 percent.

Preferably, in the detection method, if the mass spectrum of the test solution contains the compounds with the structures of formula i and formula ii, the peak areas of the compounds with the structures of formula i and formula ii are not larger than the peak areas of the compounds with the structures of formula i and formula ii in the control solution.

Preferably, for the detection method described above, wherein said lobaplatin comprises either one or both of lobaplatin diastereomer i and lobaplatin diastereomer ii.

The invention has the following beneficial effects:

the invention establishes a method for detecting platinum compounds with structures shown in formulas I and II as related substances in a lobaplatin quality standard so as to establish a lobaplatin quality detection system.

Drawings

FIG. 1 is a typical spectrum of a compound of formula I and a compound of formula II in example 1;

FIG. 2-1A is a total ion chromatogram in a white solution specificity experiment in example 4;

FIG. 2-1B is an MS chromatogram from a white solution specificity experiment in example 4;

FIGS. 2-2A are total ion chromatograms from a positional solution specificity experiment for compounds of formula I in example 4;

FIGS. 2-2B are MS chromatograms of compounds of formula I in example 4 with retention time t ═ 1.616min in a positional solution specificity experiment;

FIGS. 2-3A are total ion chromatograms from a positional solution specificity experiment for compounds of formula II in example 4;

FIGS. 2-3B are MS chromatograms with retention times t 10.984min in a positional solution specificity experiment for compounds of formula II in example 4;

FIGS. 2-4A are total ion chromatograms obtained in experiments specific for the lobaplatin test solutions of example 4;

FIGS. 2-4B are MS chromatograms of the lobaplatin test solutions of example 4 with retention time t 5.165 min;

Detailed Description

The invention provides a method for detecting a lobaplatin related substance, wherein the lobaplatin related substance comprises compounds with structures shown in formulas I and II, and the structure of the formula I is shown in the specification

The structure of the formula II is

In a preferred embodiment of the present invention, wherein the detection method is HPLC-MS method or HPLC method; preferably, the detection conditions of the HPLC-MS method are as follows: octadecylsilane chemically bonded silica is used as a filling agent, 10-12mmol/L ammonium acetate is used as a mobile phase A, and methanol: the volume ratio of acetonitrile is 1, (0.8-1.2) is used as a mobile phase B, and gradient elution is carried out; preferably, the mobile phase A is a 10mmol/L ammonium acetate solution, and the mobile phase B is methanol: the volume ratio of acetonitrile is 1: 1; preferably, the flow rate is 0.8-1.2ml, preferably 1.0ml per minute; preferably, the column temperature is 38-42 ℃, preferably 40 ℃; preferably, in the chromatogram of the system suitability test solution, the degree of separation of the peak of the substance of interest from its neighboring substances is not less than 1.5; preferably, the relative standard deviation of the peak areas of the compound with the structure of the formula I and the compound with the structure of the formula II is not more than 10.0 percent.

Preferably, for the detection method, if the compound with the structure of formula i and the compound with the structure of formula ii are present in the mass spectrum of the test solution, the peak areas of the compounds should not be larger than the peak areas (0.05%) of the compound with the structure of formula i and the compound with the structure of formula ii in the reference solution, and the 0.05% indicates that the concentration of the compound with the structure of formula i and the compound with the structure of formula ii in the reference solution is 0.05% of that of the test solution.

Herein, in the present invention, any substance affecting the purity of the drug is collectively referred to as "related substance affecting the quality of lobaplatin" or "related substance affecting the quality", and is simply referred to as "related substance", for example, a peak of related substance affecting the quality of lobaplatin appearing in an HPLC chromatogram peak for detecting the quality of lobaplatin, is simply referred to as "related substance peak"; the "related substance" in the present invention is sometimes an "impurity" known to those skilled in the art to affect the purity of the drug, however, the "related substance" in the present invention is not limited to the category of "impurity" but also includes substances having a certain anticancer activity even higher than that of lobaplatin, which belong to the category of substances related to lobaplatin with respect to the active molecule "lobaplatin", and the principles of their anticancer activity or other positive effects and functions in developing new drugs have not been fully studied.