Venoclara impurity reference substance as well as preparation method and application thereof

Abstract

The invention provides a chemical compound as shown in a formula (I) of a vonaka impurity reference substance and a synthetic route thereof. According to the invention, a brand-new impurity possibly generated in a Vickers synthesis process is obtained based on reaction conditions and reaction products of a condensation reaction through research on a synthesis route of Vickers, an impurity spectrum of Vickers is enriched and perfected, comprehensive control on the quality of Vickers is facilitated, and the quality of Vickers is improved. The yield of the vonakula is improved, and the method has important significance for optimizing the production process of the vonakula.

Description

Technical Field

[0001] The invention relates to the technical field of chemical drug analysis and drug synthesis, and in particular to a venetoclax impurity reference substance, a preparation method and an application thereof. Background Art

[0002] Venetoclax (Venetoclax), CAS registration number: 1257044-40-8, chemical name: 2-((1H-pyrrolo[2,3-B]pyridin-5-yl)oxy)-4-(4-((4'-chloro-5,5-dimethyl-3,4,5,6-tetrahydro-[1,1'-biphenyl]-2-yl)methyl)piperazin-1-yl)-N-((3-nitro-4-(((tetrahydro-2H-pyran-4-yl)methyl)amino)phenyl)sulfonyl)benzamide, its structural formula is:

[0003]

[0004] Venetoclax is a breakthrough anticancer drug developed by Abbvie and Roche. Currently, the drug is in the highest stage of development for approval and is used to treat acute myeloid leukemia and chronic lymphocytic leukemia. It is an oral B-cell lymphoma factor-2 (BCL-2) inhibitor. BCL-2 plays an important role in cell apoptosis (programmed cell death), can prevent apoptosis of some cells (including lymphocytes), and is overexpressed in certain types of cancer and is associated with the formation of drug resistance. Venetoclax is designed to selectively inhibit the function of BCL-2, restore the cell's communication system, and allow cancer cells to self-destruct to achieve the purpose of treating tumors. The drug has good clinical experience in treating various types of blood cancers, including CLL, NHL diffuse large B-cell lymphoma (DLBCL), acute myeloid leukemia (AML) and multiple myeloma (MM).

[0005] At present, the synthesis route of venetoclax is mostly based on 2-fluoro-4-bromo-1-iodobenzene as the initial raw material, and venetoclax is obtained through Grignard reaction, nucleophilic reaction, Buchwald coupling, hydrolysis, and condensation reaction. The synthesis route is as follows:

[0006]

[0007] Although some venetoclax impurities have been reported in existing literature and data, further research on other impurities that may be produced in the venetoclax synthesis process or subsequent preparation and storage processes can not only enrich and improve the impurity spectrum of venetoclax, but also has great significance for the comprehensive control of venetoclax quality. Summary of the invention

[0008] The technical problem to be solved by the present invention is to overcome the defects of the prior art. The present invention proposes a venetoclax impurity reference substance, a preparation method and an application thereof, enriches and improves the impurity spectrum of venetoclax, optimizes the venetoclax synthesis process, and improves the overall control of the quality of venetoclax.

[0009] In order to solve the above technical problems, the technical solution adopted by the present invention is: a method for preparing a venetoclax impurity reference substance, wherein the venetoclax impurity structure is shown in formula (I):

[0010]

[0011] The specific synthetic route of the venetoclax impurity is as follows:

[0012]

[0013] Furthermore, the compound of formula (II) reacts with the compound of formula (III) in the presence of a solvent and a condensing agent to obtain a compound of formula (I), wherein the molar ratio of the compound of formula (II), the compound of formula (III) and the condensing agent is 1:1-3:1.1-3.

[0014] Furthermore, the reaction temperature of the condensation reaction is 10-50°C, and the reaction time is 6-24h.

[0015] Furthermore, the condensing agent is any one of N,N'-dicyclohexylcarbodiimide, diisopropylcarbodiimide, and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride.

[0016] Furthermore, the solvent is any one of dichloromethane, dichloroethane, N,N-dimethylformamide, N,N-dimethylacetamide or tetrahydrofuran, or a mixture of several of them.

[0017] Furthermore, an activator is included, and the molar ratio between the activator and the condensing agent is 1.1-3:1.2-3, and the activator is any one of 4-dimethylaminopyridine, 1-hydroxybenzotriazole, N-hydroxysuccinimide, and pentafluorophenol.

[0018] Furthermore, a venetoclax impurity prepared according to the preparation method of the venetoclax impurity reference substance, wherein the venetoclax impurity structure is shown in formula (I):

[0019]

[0020] Furthermore, the application of a venetoclax impurity reference substance is used to enrich and improve the impurity spectrum of venetoclax and optimize the synthesis process of venetoclax.

[0021] Compared with the prior art, the beneficial effects of the present invention include: through the study of the synthesis route of venetoclax, based on the reaction conditions and reaction products of the condensation reaction, a new impurity that may be produced in the venetoclax synthesis process is obtained, which enriches and improves the impurity spectrum of venetoclax, helps to comprehensively control the quality of venetoclax, improves the yield of venetoclax, and is of great significance for optimizing the production process of venetoclax. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] The disclosure of the present invention is described with reference to the accompanying drawings. It should be understood that the drawings are for illustrative purposes only and are not intended to limit the scope of protection of the present invention. In the accompanying drawings, the same reference numerals are used to refer to the same components. Among them:

[0023] Figure 1 Schematic diagram of the synthetic route of the compound of formula (I);

[0024] Figure 2 is the HPLC spectrum of the compound of formula (I);

[0025] Figure 3 is the MS spectrum of the compound of formula (I);

[0026] Figure 4 A compound of formula (I) 13 C NMR spectrum;

[0027] Figure 5 A method for synthesizing a venetola intermediate 1 HNMR spectrum;

[0028] Figure 6 The HPLC spectrum of the crude product of Venetoclax obtained according to the synthetic route of Venetoclax;

[0029] Figure 7 HPLC spectrum of the product obtained by Buchwald coupling reaction according to the synthetic route of Venetoclax;

[0030] Figure 8 The HPLC spectrum of the crude product of Venetoclax obtained after purifying the intermediate II in the synthetic route of Venetoclax. DETAILED DESCRIPTION

[0031] It is easy to understand that according to the technical solution of the present invention, without changing the essential spirit of the present invention, a person skilled in the art can propose a variety of interchangeable structural modes and implementation modes. Therefore, the following specific implementation modes and drawings are only exemplary descriptions of the technical solution of the present invention, and should not be regarded as the whole of the present invention or as a limitation or restriction to the technical solution of the present invention.

[0032] A venetoclax impurity control substance, the synthesis route of which is as follows Figure 1 As shown, it has a structure as shown in formula (I):

[0033]

[0034] The preparation of the venetoclax impurity is described in detail below with reference to the examples.

[0035] Example 1: Preparation of the compound of formula (I)

[0036] Add the compound of formula (II) (20.0 g, 35.0 mmol, 1.0 eq) to a three-necked reaction flask, dissolve it in 300 mL of dichloromethane (DCM), and add the compound of formula (III) (11.1 g, 38.0 mmol, 1.0 eq), 4-dimethylaminopyridine (DMAP) (5.13 g, 42.0 mmol, 1.2 eq), and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI.HCl) (7.4 g, 38.5 mmol, 1.1 eq) in sequence at room temperature. Control the temperature at 20-30°C and stir the reaction for 16 hours. After the reaction is completed, the temperature is lowered, 100 mL of 5% acetic acid aqueous solution is added for washing once, and then 100 mL of 5.0% sodium bicarbonate solution is used for washing once, the organic phase is dried over 20 g of anhydrous sodium sulfate, filtered, and spin-dried to obtain a crude compound of formula (I), and the crude compound of formula (I) is crystallized from a dichloromethane / methanol system to obtain 6.01 g of a solid compound of formula (I) with a purity of 99.5% and a yield of 19.7%.

[0037] Final material testing:

[0038] The HPLC spectrum of the white solid is as follows Figure 2 shown.

[0039] The MS spectrum of the white solid is as follows Figure 3 shown.

[0040] White solid 13 C NMR spectrum Figure 4 shown.

[0041] White solid 1 HNMR spectrum Figure 5 shown.

[0042] Embodiment 2:

[0043] Add the compound of formula (II) (20.0 g, 35.0 mmol, 1.0 eq) to a three-necked reaction flask, dissolve it with 300 mL of dichloromethane (DCM), and add the compound of formula (III) (22.2 g, 70.0 mmol, 2.0 eq), 4-dimethylaminopyridine (DMAP) (6.41 g, 52.5 mmol, 1.5 eq), and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI.HCl) (10.06 g, 52.5 mmol, 1.5 eq) in sequence at room temperature. Control the temperature at 30-40°C and stir the reaction for 12 hours. After the reaction is completed, the temperature is lowered, 100 mL of 5% acetic acid aqueous solution is added for washing once, and then 100 mL of 5.0% sodium bicarbonate solution is used for washing once, the organic phase is dried over 20 g of anhydrous sodium sulfate, filtered, and spin-dried to obtain a crude compound of formula (I), and the crude compound of formula (I) is crystallized from a dichloromethane / methanol system to obtain 4.63 g of a solid compound of formula (I) with a purity of 97.2% and a yield of 15.2%.

[0044] The following is a detailed description of the application process of the venetoclax impurity reference substance. A method for preparing venetoclax is disclosed in the invention patent with publication number CN118271318 A and the name of which is a method for preparing venetoclax. The specific synthesis route is as follows:

[0045]

[0046] Wherein: R is an amino protecting group.

[0047] The specific synthesis steps are as follows:

[0048] (1) Preparation of Formula II: Formula I is used as a starting material, and reacted with an amino protecting agent, a dicarbonate diester compound, in the presence of a base to obtain Formula II;

[0049] (2) Preparation of Formula IV: Formula II and Formula III are used as raw materials and reacted under the action of a condensing agent to obtain Formula IV;

[0050] (3) Preparation of venetoclax: in the presence of a deprotecting agent, the compound of formula IV is subjected to a deprotection reaction to obtain a crude venetoclax product, which is then reacted with a sodium hydroxide methanol solution to generate venetoclax sodium salt, and finally an organic acid is added to dissolve the venetoclax sodium salt to obtain venetoclax;

[0051] The molar ratio of formula II and formula III to the condensing agent in step (2) is 1:1-2:1.1-2; and the reaction temperature is 25-40°C.

[0052] Comparing the aforementioned synthetic route of Venetoclax with the synthetic route of Venetoclax mentioned in the background technology of the present invention, it is not difficult to see that the synthetic route disclosed in this invention patent is the condensation reaction step of the synthetic route of Venetoclax mentioned in the background technology of the present invention, and the reaction conditions and the molar ratio of the relevant reactants disclosed in the invention patent are all included in the scope of the present invention, and the compound of formula (III) used as the reaction raw material in the present invention is actually the residue of Buchwald coupling in the synthetic route of Venetoclax in the present invention, so the Venetoclax impurity reference substance shown in formula (I) proposed in the present invention can actually be generated by synchronous reaction in the condensation reaction process in the synthetic route of Venetoclax in the present invention, and thus remain in the crude Venetoclax obtained in the condensation reaction, as follows Figure 6 shown.

[0053] The invention obtains a new impurity that may be generated in the synthesis process of venetoclax by studying the synthesis route of venetoclax based on the reaction conditions and reaction products of the condensation reaction, enriches and improves the impurity spectrum of venetoclax, helps to comprehensively control the quality of venetoclax, improves the yield of venetoclax, and has important significance for optimizing the production process of venetoclax; for example, in order to avoid the generation of the impurity, after the Buchwald coupling reaction is completed in the synthesis route of venetoclax, the compound of formula (III) in the reaction system is removed and then a subsequent hydrolysis and condensation reaction is performed, so that the generation and existence of the impurity in the final crude venetoclax can be effectively avoided.

[0054] In the intermediate II obtained after the Buchwald coupling reaction, a large amount of compound of formula (III) remains in the reaction system (e.g. Figure 7 ), if it is not removed, it will be derived into the finished product to form an impurity of formula I, which is difficult to remove. The crude venetoclax product needs to be purified by crystallization at least 4 times to reduce the impurity to less than 0.1%. We purified the intermediate II obtained after the Buchwald coupling reaction by formic acid / toluene crystallization to reduce the compound of formula (III) in the reaction system to about 0.3% (such as Figure 8 ), the obtained crude venetoclax only needs one crystallization to be qualified (unknown single impurity <0.1%), so the yield of venetoclax is also increased from the previous 31.7% to 38.6%, which not only reduces the number of crystallizations of the crude venetoclax and simplifies the steps of venetoclax, but also reduces the cost pressure of venetoclax by increasing the yield, making the product more competitive in the market.

[0055] The technical scope of the present invention is not limited to the contents in the above description. Those skilled in the art can make various deformations and modifications to the above embodiments without departing from the technical idea of ​​the present invention, and these deformations and modifications should all fall within the protection scope of the present invention.

Claims

1. A method for preparing a venetoclax impurity reference substance, characterized in that: The structure of the venetoclax impurity is shown in formula (I): The specific synthetic route of the venetoclax impurity is as follows:

2. The method for preparing the venetoclax impurity reference substance according to claim 1, characterized in that: The compound of formula (II) reacts with the compound of formula (III) in the presence of a solvent and a condensing agent to obtain a compound of formula (I), wherein the molar ratio of the compound of formula (II), the compound of formula (III) and the condensing agent is 1:1-3:1.1-3.

3. The method for preparing the venetoclax impurity reference substance according to claim 2, characterized in that: The reaction temperature of the condensation reaction is 10-50° C., and the reaction time is 6-24 hours.

4. The method for preparing the venetoclax impurity reference substance according to claim 3, characterized in that: The condensing agent is any one of N,N'-dicyclohexylcarbodiimide, diisopropylcarbodiimide, and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride.

5. The method for preparing the venetoclax impurity reference substance according to claim 3, characterized in that: The solvent is any one of dichloromethane, dichloroethane, N,N-dimethylformamide, N,N-dimethylacetamide or tetrahydrofuran, or a mixture of the two or more thereof.

6. The method for preparing the venetoclax impurity reference substance according to claim 3, characterized in that: The invention also comprises an activator, wherein the molar ratio between the activator and the condensing agent is 1.1-3:1.2-3, and the activator is any one of 4-dimethylaminopyridine, 1-hydroxybenzotriazole, N-hydroxysuccinimide and pentafluorophenol.

7. A venetoclax impurity prepared according to the method for preparing a venetoclax impurity reference substance according to any one of claims 1 to 6, characterized in that: The structure of the venetoclax impurity is shown in formula (I):

8. An application of a venetoclax impurity reference substance, characterized in that: Used to enrich and improve the impurity spectrum of Venetoclax and optimize the synthesis process of Venetoclax.

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