CN117924163A

CN117924163A - Impurity compound of isaconazole onium sulfate and preparation method thereof

Info

Publication number: CN117924163A
Application number: CN202311833128.1A
Authority: CN
Inventors: 孟磊; 顾理群; 周西朋; 龚彦春; 刘云龙
Original assignee: Hubei Tianshu Pharmaceutical Co ltd; Nanjing Weikaier Biomedical Technology Co ltd
Current assignee: Hubei Tianshu Pharmaceutical Co ltd; Nanjing Weikaier Biomedical Technology Co ltd
Priority date: 2023-12-28
Filing date: 2023-12-28
Publication date: 2024-04-26

Abstract

The invention belongs to the technical field of medicines, and particularly relates to an impurity compound of isaconazole onium sulfate and a preparation method thereof. The invention discovers a new genotoxic impurity VI, has good warning effect on the process optimization of the step to reduce the generation of the impurity, is also beneficial to the detailed study on the removal and transmission of the impurity in the subsequent synthesis of the isaconazole onium sulfate bulk drug, and avoids the transmission of the impurity into the bulk drug. The invention also provides a preparation method of the impurity VI, which is used as an impurity reference substance of the isaconazole onium sulfate or the pharmaceutical composition thereof, and the preparation method of the invention solves the problem of mass production of the impurity reference substance, and has the advantages of high yield, low three wastes, low cost and high production efficiency.

Description

Impurity compound of isaconazole onium sulfate and preparation method thereof

Technical Field

The invention belongs to the technical field of medicines, and particularly relates to an impurity compound of isaconazole onium sulfate and a preparation method thereof.

Background

Adverse reactions occurring in clinical use of a drug are sometimes associated with impurities present in the drug, in addition to the pharmacological activity of the drug itself. Therefore, the impurity research is performed regularly, and the quality and the safety of the medicines on the market are directly related.

The isaconazole onium sulfate (BAL 8557-002) is a triazole antifungal drug isaconazole prodrug, which is metabolized into isaconazole to exert antifungal action mechanism after entering the body. The U.S. FDA approved priority for isaconazole onium sulfate (Isavuconazonium Sulfate) from An Si thai, japan, 3-month 6, commercially available under the trade name: cresemba for the treatment of invasive aspergillosis and mucormycosis.

The synthesis method of the isaconazole onium sulfate is generally that the isaconazole mother nucleus A and a side chain B are condensed, and then a series of reactions are carried out, wherein the structures of the mother nucleus A and the side chain B are as follows:

The compound of formula VII, as a key intermediate for the synthesis of side chain B, can be prepared by reacting a compound of formula VIII with a compound of formula IX:

structurally, compounds of formulas VII and VIII both have the type sa_28bis:Aromatic mono-AND DIALKYLAMINE suspected genotoxicity alerting structure described in Romualdo Benigni,et al.Development of structural alerts for the in vivo micronucleus assay in rodents.JRC Scientific and Technical Reports2009:

Although compounds of formulas VII and VIII have been studied as suspected genotoxic impurities, the risk of the presence of other genotoxic impurities that may be introduced into the product is still relatively large, and the presence of unidentified suspected genotoxic impurities can have a significant adverse effect on the quality of isaconazole onium sulfate or a pharmaceutical composition thereof. At present, no report is made in the prior art regarding genotoxic impurities that may be produced by formula VIII in the reaction for preparing the compounds of formula VII. Therefore, in order to ensure the product quality of the isaconazole onium sulfate, it is important to identify the reaction byproducts in the reaction and whether unknown suspected genotoxic impurities exist in the side reaction products.

Disclosure of Invention

The inventors found in the analytical study of the synthesis of the compound of formula VII,

LC-MS showed the presence of an impurity having a molecular weight of 429.3 in the product of the compound of formula VII.

The LC-MS method is as follows:

Chromatographic column: yuehuan Xtimate C, 50X 4.6mm,3 μm

Detection wavelength: column temperature of 210nm/254 nm: flow rate at 40 ℃): sample injection amount of 1.0 mL/min: 1 mu L

Mobile phase a: acetonitrile mobile phase B:0.1% formic acid water

The mobile phase elution procedure was as follows:

the mass spectrometry conditions were as follows:

mode: full scan

Molecular weight range: 100 to 1000

Fragment generation voltage: 70

Speed of: 1733 u/sec

Drying gas flow rate: 12.0L/min

Atomizer pressure: 35psig

Drying gas temperature: 350 DEG C

Capillary voltage: and (3) a positive electrode: 5000V, negative electrode: 3000V

The retention times of LC-MS are shown in Table 1 and the mass spectrum is shown in FIG. 1.

TABLE 1

Retention time (min)	[M+1]	Remarks
			3.55	310.1	Compounds of formula VII
4.25	430.3	Unknown impurities

The applicant has carried out research and analysis on the reaction mechanism, and has presumed that the generation mechanism of the unknown impurity with the molecular weight of 429.3 is as follows: the resulting product of the reaction, the compound of formula VII, continues to react with the starting compound of formula VIII, thereby producing the impurity of formula VI.

The present invention has been completed based on the above-described studies.

In one aspect, the invention provides an impurity VI of isaconazole onium sulfate:

In another aspect, there is provided the use of impurity VI as an impurity control in quality control of isaconazole onium sulfate or a pharmaceutical composition thereof.

The structure of impurity VI is considered to have a suspected genotoxicity warning structure of SA_28bis:Aromatic mono-AND DIALKYLAMINE type described in Romualdo Benigni,et al.Development of structural alerts for the in vivo micronucleus assay in rodents.JRC Scientific and Technical Reports 2009:

The applicant therefore further carried out genotoxicity studies carried out with the experimental kit using the genetics toxicity Ames kit-5 version of Hui Zhi and Source biotechnology (Suzhou) limited, the strains TA97a, TA98, TA100, WP2uvrA pKM101 and TA1535 respectively, the experimental methods were carried out with reference to the guidelines of the economic Cooperation and development organization "OECD 471:Bacterial Reverse Mutation Test" and GB15193.4-2014, the experimental results showed that: at a dose concentration of 0.5ug/plate, the test results of the five strains were positive, proving that impurity VI was genotoxic impurity.

Considering the genotoxic impurity property of the impurity VI, in the drug research, the transfer and removal process of the impurity VI in the synthesis of the bulk drug needs to be examined, a certain amount of impurity VI is needed, and the impurity VI is simply enriched through the preparation liquid phase, so that the problems of low efficiency, large amount of waste liquid and the like are solved, and therefore, the invention also provides a preparation method of the impurity VI of the isaconazole onium sulfate:

Step (3): removing Boc protecting groups from the compound of the formula III under an acidic condition to prepare a compound of the formula IV;

step (4): the compound of formula IV reacts with the compound of formula V under the action of an alkaline reagent, a catalyst and a condensing agent to obtain an impurity VI.

Further, the acidic conditions in step (3) are selected from hydrochloric acid, trifluoroacetic acid or methylsulfonic acid; the reaction solvent is selected from methanol, ethanol, ethyl acetate or 1, 4-dioxane; the reaction temperature is 20-40 ℃.

Further, in step (4), the basic reagent is selected from triethylamine or N, N-diisopropylethylamine; the catalyst is selected from 4-dimethylaminopyridine; the condensing agent is selected from 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride or 2- (7-azobenzotriazole) -N, N, N ', N' -tetramethyl urea hexafluorophosphate; the reaction temperature is 0-5 ℃.

Still further, the compound of formula III may be prepared by a compound of formula I:

Step (1): reacting the compound of the formula I with Boc anhydride under alkaline conditions to prepare a compound of the formula II;

step (2): and (3) carrying out heating reaction on the compound of the formula II and manganese dioxide in 1, 2-dichloroethane to prepare the compound of the formula III.

Further, the basic conditions in step (1) are selected from triethylamine, N-diisopropylethylamine, potassium carbonate or sodium hydroxide; the reaction solvent is selected from dichloromethane, tetrahydrofuran, 1, 4-dioxane, toluene, methanol, ethanol or N, N-dimethylformamide. Still further, a catalyst, 4-dimethylaminopyridine, is preferably used in the step (1).

Further, the heating reaction temperature in the step (2) is 60-65 ℃; the heating reaction time is 15-20 hours.

The preparation method of the impurity VI of the isaconazole onium sulfate is preferably as follows:

Step (2): heating a compound of the formula II and manganese dioxide in 1, 2-dichloroethane to react and prepare a compound of the formula III;

The basic conditions in step (1) are selected from triethylamine, N-diisopropylethylamine, potassium carbonate or sodium hydroxide; the reaction solvent is selected from dichloromethane, tetrahydrofuran, 1, 4-dioxane, toluene, methanol, ethanol or N, N-dimethylformamide. In step (1) it is also preferred to use a catalyst which is 4-dimethylaminopyridine.

The heating reaction temperature in the step (2) is 60-65 ℃; the heating reaction time is 15-20 hours.

The acidic conditions in step (3) are selected from hydrochloric acid, trifluoroacetic acid or methanesulfonic acid; the reaction solvent is selected from methanol, ethanol, ethyl acetate or 1, 4-dioxane; the reaction temperature is 20-40 ℃.

The basic reagent in step (4) is selected from triethylamine or N, N-diisopropylethylamine; the catalyst is selected from 4-dimethylaminopyridine; the condensing agent is selected from 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride or 2- (7-azobenzotriazole) -N, N, N ', N' -tetramethyl urea hexafluorophosphate; the reaction temperature is 0-5 ℃.

The beneficial effects are that:

The invention discovers a new genotoxic impurity VI, has good warning effect on the process optimization of the step to reduce the generation of the impurity, is also beneficial to the detailed study on the removal and transmission of the impurity in the subsequent synthesis of the isaconazole onium sulfate bulk drug, and avoids the transmission of the impurity into the bulk drug.

The invention also provides a preparation method of the impurity VI, which solves the problem of mass production of the impurity control product, and has the advantages of high yield, low three wastes, low cost and high production efficiency.

Drawings

Mass spectra of fig. 1, compounds of formula VII and impurity VI.

FIG. 2 shows ¹ H-NMR nuclear magnetic spectrum of impurity VI.

Fig. 3, high performance liquid chromatogram of impurity VI.

FIG. 4, system adaptation diagram of methods for liquid phase purity detection of compounds of formula VII and impurity VI.

Detailed Description

The invention is described in more detail below by means of examples. However, these specific descriptions are only for illustrating the technical scheme of the present invention, and do not limit the present invention in any way.

Example 1

Genotoxicity study experiment of impurity VI

The experimental kit adopts a genetics toxicity Ames kit-5 bacteria plate of Hui Zhi and Source biotechnology (Suzhou) limited company, strains are TA97a, TA98, TA100, WP2uvrApKM101 and TA1535 strains respectively, and the experimental method is carried out by referring to guidelines of economic Cooperation and development organization (OECD 471:Bacterial Reverse Mutation Test) and GB 15193.4-2014: the experiment adopts a flat plate infiltration method, adopts DMSO to dissolve impurity VI, and designs the following groups of experiments with dosages respectively: 0. 0.1, 0.25, 0.5, 1, 2.5, 5, 7.5ug/plate. The concentrations of impurity VI for each dose were 0, 0.04, 0.09, 0.19, 0.37, 0.93, 1.85, 2.78ug/mL, respectively, as shown in table 2.

TABLE 2

Dosage (ug/plate)	Impurity VI concentration (ug/mL)
		0	0
0.1	0.04
		0.25	0.09
0.5	0.19
		1	0.37
2.5	0.93
		5	1.85
7.5	2.78

The experimental results show that: at a dose concentration of 0.5ug/plate, the test results of the five strains were positive, proving that impurity VI was genotoxic impurity.

Example 2

Synthesis of Compounds of formula II

To the reaction flask were added the compound of formula I (200 g), methylene chloride (2L), 4-dimethylaminopyridine (53 g) and triethylamine (439.5 mL), followed by slow dropwise addition (Boc) ₂ O (379 g) at 0℃and stirring at room temperature for 4 hours. After the reaction, water (2L) was added, the layers were separated, the aqueous phase was extracted once with dichloromethane (1L), the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give crude compound of formula II as a yellow oil, 345g, which was directly used in the next reaction.

Example 3

Synthesis of Compound of formula III

To the reaction flask were added the compound of formula II (345 g), 1, 2-dichloroethane (3.45L) and manganese dioxide (1259 g), and the reaction was stirred at 60 to 65℃for 16 hours. After the reaction, cooling to room temperature, filtering with diatomite, flushing a filter cake with methanol (1L), concentrating the filtrate under reduced pressure to obtain a crude product, and purifying the crude product by column chromatography (PE: EA=5:1) to obtain 63g of pure compound of the formula III, wherein the yield is 12.14%.

Example 4

Synthesis of Compound of formula IV

To the reaction flask were added the compound of formula III (63 g) and methanol (630 mL), and hydrochloric acid (25.6 g) was added dropwise at 0℃under controlled temperature, and the mixture was stirred at room temperature for 1 hour. After the reaction, the pH is regulated to 7-8 by using a sodium hydroxide aqueous solution, dichloromethane (1L) and water (0.8L) are added, the layers and the liquid are separated, the aqueous phase is extracted for 2 times by using dichloromethane, the organic phases are combined, dried by using anhydrous sodium sulfate and concentrated under reduced pressure to obtain 43g of the compound shown in the formula IV, and the yield is 94.70%.

Example 5

Synthesis of Compounds of formula VI

To the reaction flask were added the compound of formula IV (43 g), the compound of formula V (157 mL), 4-dimethylaminopyridine (6.1 g), and methylene chloride (430 mL), N-diisopropylethylamine (64.5 g) was added dropwise to the reaction at 0℃and then 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (38.3 g). After the reaction, an ammonium chloride aqueous solution (300 mL) was added, the layers were separated, the organic phase was washed with water for 1 time, the aqueous phase was combined, extracted with dichloromethane for 1 time, the organic phase was combined, dried over anhydrous sodium sulfate, concentrated under reduced pressure to give 80g of a crude compound of formula VI, purified by preparative HPLC, and the preparation was lyophilized to give 22.5g of a pure compound of formula VI in a yield of 31.47% and a purity of 97.24%.

Example 6

Impurity VI was developed as an analytical method for impurity controls for compounds of formula VII

The HPLC analysis method is as follows:

Chromatographic column: YMC TRIART PFPC.times.18X14.6mm, 5 μm

Ghost Sniper 50×4.6μm

Detection wavelength: column temperature 240 nm: flow rate at 25 ℃): sample injection amount of 1.0 mL/min: 10 μl mobile phase a: acetonitrile: methanol=9:1

Mobile phase B:5mMol Potassium hexafluorophosphate (pH 2.6.+ -. 0.1)

The mobile phase elution procedure was as follows:

The retention times for HPLC are shown in table 3 and the system applicability profile is shown in fig. 4.

TABLE 3 Table 3

The impurity VI and the compound of the formula VII have better separation degree by adopting the analysis method.

Claims

1. An impurity (VI) of isaconazole onium sulfate,

2. A preparation method of impurities (VI) of isaconazole onium sulfate,

The method is characterized in that:

Step (3): removing Boc protecting groups from the compound of the formula (III) under an acidic condition to prepare a compound of the formula (IV);

step (4): the compound of formula (IV) reacts with the compound of formula (V) under the action of an alkaline reagent, a catalyst and a condensing agent to obtain the impurity (VI).

3. The process for the preparation of impurities (VI) of isaconazole onium sulphate according to claim 2, wherein: the acidic conditions in step (3) are selected from hydrochloric acid, trifluoroacetic acid or methanesulfonic acid; the reaction solvent is selected from methanol, ethanol, ethyl acetate or 1, 4-dioxane; the reaction temperature is 20-40 ℃.

4. The process for the preparation of impurities (VI) of isaconazole onium sulphate according to claim 2, wherein: the basic reagent in step (4) is selected from triethylamine or N, N-diisopropylethylamine; the catalyst is selected from 4-dimethylaminopyridine; the condensing agent is selected from 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride or 2- (7-azobenzotriazole) -N, N, N ', N' -tetramethyl urea hexafluorophosphate; the reaction temperature is 0-5 ℃.

5. The process for the preparation of impurities (VI) of isaconazole onium sulphate according to claim 2, wherein: the compound of formula (III) is prepared from a compound of formula (I):

Step (1): reacting a compound of formula (I) with Boc anhydride under alkaline conditions to prepare a compound of formula (II);

Step (2): the compound of the formula (II) and manganese dioxide are heated and reacted in 1, 2-dichloroethane to prepare the compound of the formula (III).

6. The process for producing the impurity (VI) of isaconazole onium sulfate according to claim 5, wherein: the basic conditions in step (1) are selected from triethylamine, N-diisopropylethylamine, potassium carbonate or sodium hydroxide; the reaction solvent is selected from dichloromethane, tetrahydrofuran, 1, 4-dioxane, toluene, methanol, ethanol or N, N-dimethylformamide.

7. The process for the preparation of impurities (VI) of isaconazole onium sulfate according to claim 6, wherein: and (2) a catalyst is also used in the step (1), and the catalyst is 4-dimethylaminopyridine.

8. The process for producing the impurity (VI) of isaconazole onium sulfate according to claim 5, wherein: the heating reaction temperature in the step (2) is 60-65 ℃; the heating reaction time is 15-20 hours.

9. The process for the preparation of impurities (VI) of isaconazole onium sulphate according to any one of claims 2-8, wherein:

Step (2): heating a compound of formula (II) and manganese dioxide in 1, 2-dichloroethane to obtain a compound of formula (III);

10. The use of impurity (VI) for quality control in isaconazole sulphate or a pharmaceutical composition thereof,

The method is characterized in that: the impurity (VI) is used as an impurity reference substance in the isaconazole sulfate or the pharmaceutical composition thereof.