WO2020181686A1

WO2020181686A1 - Preparation method for drug-linker mc-mmaf used for antibody drug conjugates and intermediate thereof

Info

Publication number: WO2020181686A1
Application number: PCT/CN2019/092948
Authority: WO
Inventors: 许喆; 李海泓; 郭茂君; 李辉
Original assignee: 联宁（苏州）生物制药有限公司
Priority date: 2019-03-08
Filing date: 2019-06-26
Publication date: 2020-09-17
Also published as: CN109824759A

Abstract

The present invention provides a preparation method for a drug-linker MC-MMAF used for antibody drug conjugates and an intermediate thereof. The preparation method of the present invention improves the N-terminal reactivity and thus effectively controls the racemization reaction. A compound having low toxicity is used instead of toxin MMAF so as to reduce the operation difficulty in mass production. No reverse phase preparation is required, therefore the operation is simple.

Description

Preparation method and intermediate of drug-linker MC-MMAF for antibody drug conjugate

Technical field

The invention relates to the field of organic synthesis, in particular to a preparation method and intermediates of a drug-linker MC-MMAF used for antibody drug conjugates.

Background technique

Antibody drug conjugate (ADC for short) is a new type of anti-tumor drug. Its principle is to link cytotoxin to antibody. Through antibody recognition of specific antigen on the surface of cancer cell, it enters the cancer through endocytosis. Cells, thereby transporting cytotoxins to the target, achieve the purpose of targeted treatment of malignant tumors. Compared with traditional small-molecule anti-tumor drugs, ADC is more specific and effective because it can rely on the target recognition of antibodies and the high activity of toxins.

ADC includes three different components, namely antibody, linker and cytotoxin. The antibody achieves targeting, and the linker ensures the stability of the ADC during blood transport. After reaching the target point, the toxin exerts a killing effect on cancer cells. According to different mechanisms of action, toxins suitable for ADC are classified into microtubule inhibitors, DNA damaging agents, RNA polymerase inhibitors, and so on. At present, the toxins used by ADCs sold in the market and in clinical trials are mainly microtubule inhibitors, mainly including dolastatin-based compounds, such as MMAE, MMAF and MMAD, and maytansine-based (Maytansine-based) designed compounds, such as DM1 and DM4. In terms of linkers, the main applications are non-cleavable types, such as valine-citrulline (Valine-Citriline) and cyclohexyl carboxylic acid (MCC). After lysosomal hydrolysis, the drug still has activity, and is connected to the A certain amino acid residue is joined together.

There are many ways to form antibody-drug conjugates. Either the amino or sulfhydryl group on the antibody and the drug linker can be chemically coupled, or the antibody can be modified. After a specific functional group is introduced on the antibody, it can be coupled with the drug linker for chemical reaction or enzymatic reaction coupling. United. The structure of the antibody drug conjugate MC-MMAF involved in the present invention is shown below.

The synthesis route of MC-MMAF currently reported in the literature is to use the toxin MMAF and MC-hex-Acid (1-maleimido n-hexanoic acid) to undergo a dehydration reaction to obtain MC-MMAF. The structure of MMAF is:

The synthesis scheme reported in the literature is:

The N-terminal valine of this route of MMAF has a methyl group on the N, which is sterically hindered. In this case, the reaction speed of connecting 1-maleimido n-hexanoic acid to MMAF will be slower. Even if different amide condensing agents are used, it will cause racemization of the chiral carbon linked to the phenylpropionamide group of MMAF. This route is used for the synthesis of MC-MMAF of less than 1g, and finally high-pressure reverse phase preparation is used to remove isomeric impurities, and the yield is less than 50%.

This reaction route shows certain defects during scale-up production, such as: 1. Because the condensing agent will activate the carboxyl group on MMAF at the same time, this method will cause 30-50% racemization, forming difficult to remove isomer impurities, and affect Yield; 2. Due to the aforementioned steric hindrance, the reaction time is long, and the impurities are many, which cause difficulties in the post-treatment and purification of the reaction; 3. The final product needs to be prepared by high-pressure reverse phase to remove isomers, which increases operating costs; 4. Direct Using the toxin MMAF as a raw material, it is necessary to do a good job of protection in a large number of synthesis operations, and the selection of protective equipment will bring obstacles to production operations.

Summary of the invention

On the one hand, in view of the defects of the prior art, the present invention provides a method for synthesizing MC-MMAF. The key to the reaction is to use the structural formula as

Compound with L-phenylalanine

The condensation reaction directly obtains MC-MMAF or its salt, and R is selected from one or more of hydrogen, succinimidyl, pentafluorophenyl, p-nitrophenyl, and phthalamide.

The above objectives of the present invention are achieved by the following technical solutions.

The synthesis method includes the following steps: 1) The compound

Dissolve in a suitable solvent, and

The amide condensation reaction takes place to obtain MC-MMAF.

Preferably, in step 1), the suitable solvent is selected from dichloromethane, dimethyl sulfoxide, N,N-dimethylformamide, N,N-dimethylacetamide, tetrahydrofuran, 1, One or more of 4-dioxane and 2-methyltetrahydrofuran; more preferably, the suitable solvent is selected from one or more of dichloromethane and N,N-dimethylformamide Kind.

Preferably, in step 1), if R is hydrogen, reagent N is added under the action of reagent M, which is selected from the group consisting of DCC, DCEP, EDC, DIC, HATU, HBTU, HBPIPU, HBPyU, HSPyU, HCTU, HOTU, HOTT, HSTU, HDMA, TATU, TBTU, TCTU, TCFH, TDBTU, TOTU, TOTT, TPTU, TFFH, BTFFH, TNTU, TSTU, COMU, T3P, BOP, PyBOP, PyBrOP, PyClOP, BroP, PyAOP, One or more of PyCIU, CDI, TPSI, TSTU, DEPBT, DMTMM, EEDQ, CIP, CIB, DMC, HOBt and EDCI; more preferably, the reagent M is selected from EDCI, EDC, DIC, HOAt and HOBt One or more of them; further preferably, the reagent M is a mixture of EDCI, EDC or DIC and HOAt or HOBt; most preferably, the reagent M is a mixture of EDCI and HOBt. The reagent N is selected from triethylamine, diisopropylethylamine (DIEA), pyridine, N,N-dimethyl-4-pyridine, preferably diisopropylethylamine (DIEA). The reaction temperature of the reaction is -20°C to 40°C, preferably -10°C to 25°C.

Preferably, in step 1), if R is one or more of perimidyl, pentafluorophenyl, p-nitrophenyl, and phthalamide, under the action of reagent P, versus

A reaction occurs and MC-MMAF is obtained. The reagent P is selected from triethylamine, diisopropylethylamine (DIEA), pyridine, N,N-dimethyl-4-pyridine, sodium carbonate, sodium bicarbonate, potassium carbonate, potassium bicarbonate, lithium carbonate And one or more of lithium bicarbonate, preferably sodium carbonate, or diisopropylethylamine (DIEA). The reaction temperature is 0°C to 100°C, preferably 15°C to 50°C.

Preferably, in step 1), it further includes a step of separating MC-MMAF from the reaction liquid after the reaction is completed.

Preferably, the separation includes evaporating the solvent under reduced pressure, and then purifying or recrystallization by medium pressure chromatography to obtain MC-MMAF.

The preparation method of the present invention improves the reactivity of the N terminal, thereby effectively controlling the occurrence of racemization; instead of directly using the toxin MMAF, it uses the less toxic L-phenylalanine, which reduces the operation difficulty in mass production ; No need for reverse phase preparation, simple operation. As mentioned above, this method reduces the difficulty of operation, makes the quality standard easier to control, and can be applied to the preparation of one hundred grams.

On the other hand, this patent also provides an intermediate compound for the synthesis of MC-MMAF, the structural formula of which is

Wherein, R is selected from one of hydrogen, succinimidyl, pentafluorophenyl, p-nitrophenyl, and phthalamide. It is preferably the following compound, as shown in Table 1:

Table 1

In another aspect, the present invention also provides a synthetic

The method of synthesis is as follows,

This route contains the important intermediate H for synthesis, and there is no report on the synthesis method of this compound before.

To synthesize compound H, the usual idea is to use the following route:

However, experiments have found that H1 will decompose in the process of removing tert-butyl, and the yield is very low.

If the carboxyl group is not protected, the carboxyl group of compound F will also participate in the reaction during the condensation process, resulting in a complicated reaction and many by-products.

In the method adopted in the present invention, the compound G, HATU and DIEA are first stirred and reacted at room temperature under nitrogen protection for 30 minutes, and then compound F is added and reacted under nitrogen protection at room temperature and stirred for 4 hours. Through repeated attempts to improve the feeding sequence and feeding ratio, the carboxyl group of compound F does not participate in the reaction, and compound H can be obtained in a high yield. By changing the reaction conditions, the yield of compound H is greatly improved, and the route has the possibility of being applied to production.

The present invention abandons the existing MMAF synthesis route and regards MC-MMAF as a whole for synthesis. The biggest problem is that MC linkers are fragments with relatively high reactivity. Connecting MC in advance will increase the difficulty of synthesis. Those skilled in the art would not think of this route. Through many studies, we have solved the problem of instability in the synthesis of the MC fragment compound introduced in advance, so that this overall synthetic route can be realized.

As used in this article, the definitions of commonly used organic abbreviations and their corresponding CAS numbers are shown in Table 2:

Table 2

Description of the drawings

Figure 1 is a high performance liquid chromatogram of DMT-3 synthesized in the present invention.

Figure 2 is a liquid chromatogram of compound A synthesized in the present invention.

Figure 3 is a mass spectrum of compound A synthesized in the present invention.

Figure 4 is a liquid chromatogram of compound C synthesized in the present invention.

Figure 5 is a mass spectrum of compound C synthesized in the present invention.

Figure 6 is a liquid chromatogram of compound E synthesized in the present invention.

Figure 7 is a mass spectrum of compound E synthesized in the present invention.

Figure 8 is a liquid chromatogram of compound F synthesized in the present invention.

Figure 9 is a mass spectrum of compound F synthesized in the present invention.

Figure 10 is a liquid chromatogram of compound H synthesized in the present invention.

Figure 11 is a mass spectrum of compound H synthesized in the present invention.

Figure 12 is a liquid chromatogram of the target product MC-MMAF synthesized in Condensation Scheme 1 of the present invention.

Figure 13 is a mass spectrum of the target product MC-MMAF synthesized in Condensation Scheme 1 of the present invention.

Figure 14 is the NMR spectrum of the target product MC-MMAF synthesized in Condensation Scheme 1 of the present invention.

Figure 15 is a high performance liquid chromatogram of the target product MC-MMAF synthesized by the condensation scheme 1 of the present invention.

Figure 16 is a liquid chromatogram of compound J synthesized in the present invention.

Figure 17 is a mass spectrum of compound J synthesized in the present invention.

Figure 18 is a liquid chromatogram of the target product MC-MMAF synthesized in Condensation Scheme 1 of the present invention.

Figure 19 is a mass spectrum of the target product MC-MMAF synthesized in Condensation Scheme 1 of the present invention.

detailed description

The technical solution of the present invention will be further described in non-limiting detail below in conjunction with specific embodiments. It should be pointed out that the following embodiments are only to illustrate the technical concept and features of the present invention, and their purpose is to enable those familiar with the technology to understand the content of the present invention and implement them accordingly, and cannot limit the protection of the present invention. range. All equivalent changes or modifications made according to the spirit of the present invention should be covered by the protection scope of the present invention.

LCMS means liquid-mass spectrometry detection method; HPLC means high-performance liquid chromatography detection.

The raw materials and reagents for each step of the reaction involved in the present invention can be purchased from the market or prepared according to the method of the present invention.

The present invention provides a method for synthesizing MC-MMAF, which includes the following steps:

1) The compound

Dissolved in a suitable solvent, the suitable solvent is selected from dichloromethane, dimethyl sulfoxide, N,N-dimethylformamide, N,N-dimethylacetamide, tetrahydrofuran, 1,4- One or more of dioxane and 2-methyltetrahydrofuran, and

The amide condensation reaction takes place to obtain MC-MMAF.

In step 1), if R is hydrogen, under the action of reagent M, add reagent N, which is selected from DCC, DCEP, EDC, DIC, HATU, HBTU, HBPIPU, HBPyU, HSPyU, HCTU, HOTU , HOTT, HSTU, HDMA, TATU, TBTU, TCTU, TCFH, TDBTU, TOTU, TOTT, TPTU, TFFH, BTFFH, TNTU, TSTU, COMU, T3P, BOP, PyBOP, PyBrOP, PyClOP, Brop, PyAOP, PyCIU, CDI , TPSI, TSTU, DEPBT, DMTMM, EEDQ, CIP, CIB, DMC, HOBt and EDCI; more preferably, the reagent M is selected from one of EDCI, EDC, DIC, HOAt and HOBt One or more; further preferably, the reagent M is a mixture of EDCI, EDC or DIC and HOAt or HOBt; most preferably, the reagent M is a mixture of EDCI and HOBt. The reagent N is selected from triethylamine, diisopropylethylamine (DIEA), pyridine, N,N-dimethyl-4-pyridine, preferably diisopropylethylamine (DIEA). The reaction temperature is -20°C to 40°C, preferably -10°C to 25°C.

In step 1), if R is one or more of perimido group, pentafluorophenyl group, p-nitrophenyl group, and phthalamide group, under the action of reagent P, and

In step 1), it also includes the step of separating MC-MMAF from the reaction solution after the reaction is completed. Preferably, the separation includes evaporating the solvent under reduced pressure, and then purifying or recrystallization by medium pressure chromatography to Get MC-MMAF.

The invention also provides a synthetic

Method, first put the compound

HATU and DIEA are stirred at room temperature under nitrogen protection, and then the compound

The reaction was carried out by stirring at room temperature under nitrogen protection.

Example 1

The reaction route of this example is as follows:

Condensation scheme 1

Condensation Scheme 2

In a 3L three-necked flask, add 1.5L of dichloromethane and Dil.HCl (202.3g, 0.683mol 1.0eq), magnetic stirring, nitrogen protection, and then add Z-Val-OH (163.23g, 0.65mol, 0.95eq) and HATU (311.6g, 0.82mol, 1.20eq), stirred at room temperature for 30 minutes, and then cooled to an ice bath. Add DIEA (452.5ml, 4.0eq) dropwise at 10 degrees Celsius. After the addition is complete, stir under ice bath for 30 minutes, then move to The reaction was carried out at room temperature for 16 hours, and detected by HPLC. The main peak was the product peak (retention time 29.98min). The reaction of the raw material Dil.HCl was completed and the reaction was completed. The reaction solution was washed with citric acid aqueous solution (2L*1), saturated sodium bicarbonate solution (2L*1), saturated brine (2L*1), the organic layer was dried with anhydrous sodium sulfate, filtered with suction, and desolventized 531g of crude product was obtained. The crude product was dissolved in 800ml methanol, 1.1ml (1mol/L) dilute hydrochloric acid (about 1 hour) was added dropwise with stirring in an ice bath, stirred at room temperature for 12h, stirring was stopped, the layers were separated, the upper aqueous layer was separated, and the lower product was The oil pump is pulled dry, and 325g DMT-1 is obtained, the yield is 91%

In a 2L single-mouth flask, add 800ml methanol and DMT-1 (LN114-38,325g, 0.66mol) and 110g Pd(OH)2/C, replace with H2 three times, react at room temperature for 5 hours, and TLC monitors the complete reaction of the raw material DMT-1. Add diatomaceous earth to the sand core funnel, filter with suction, and wash the filter cake with 1L methanol, collect the filtrate, evaporate the filtrate, and pump until the product does not foam, obtain DMT-2 230.2g, purity 94%; yield: 97 %.

In a 3L three-neck flask, dissolve DMT-2 (LN114-40-01, 230.2g, actual 0.60mol, 1.0eq) in 500ml DCM, stir well, and add Fmoc-Me-val (202.6g, 0.57mol, 0.95 eq) and HATU (292.9g, 0.77mol, 1.20eq), then add 1L DCM, stir at room temperature for 30min, then cool to an ice bath, control the temperature at 10 degrees and add DIEA (212.7ml, 2.0eq) dropwise, after the addition is complete, ice bath After stirring for 30 min, it was moved to room temperature and reacted for 16.0 h. HPLC detected the main peak as the product peak (retention time 36.00 min), the reaction of the raw material DMT-2 was complete, and the reaction ended. The reaction solution was washed with water (2.0L*1), citric acid aqueous solution (2L*1), saturated sodium bicarbonate solution (2L*1), saturated brine (1L*1), and the organic layer was washed with anhydrous sulfuric acid After the sodium was dried, filtered with suction to remove the solvent to obtain a crude product of 655g. The crude product was dissolved in 650ml of methanol, and 360ml (1mol/L) of dilute hydrochloric acid was added dropwise with stirring. The stirring was stopped for 12h at room temperature. The stirring was stopped, the layers were separated, and the upper aqueous layer was separated. all over. The lower product was pulled dry by an oil pump to obtain 373g DMT-3 with a HPLC purity of 96.7% and a yield of 90%.

Compound DMT-3 (173g, 0.25mol) was added to dichloromethane (500mL) and fully stirred, trifluoroacetic acid (170mL) was added under ice bath, slowly warmed to room temperature under nitrogen protection, stirred and reacted for 16 hours, LCMS showed the reaction liquid Less than 1% of the compound DMT-3 is regarded as the end of the reaction. The reaction solution was washed with water (500mL×3) and then with saturated brine (500mL). The organic phase was dried with anhydrous Na ₂ SO ₄ and concentrated on a rotary evaporator to obtain a pale yellow crude product 165g; the crude product was added to 1.1L (petroleum ether) : Ethyl acetate = 10:1) In the mixed solvent with mechanical stirring, a white solid will precipitate out after about 1 hour. After filtration, the filter cake is washed twice with DCM (500 mL) and dried under vacuum to obtain compound A (136g, yield 85%, ). MS: 638.27 (M+H ⁺ )

Compound A (2.38 g, 3.71 mmol), compound B (456 mg, 3.96 mmol) and compound EDCI (760 mg, 3.96 mmol) were dissolved in dichloromethane (20 mL), and the reaction was stirred at room temperature under nitrogen protection for 2 hours. LCMS showed the reaction solution Less than 5% of the compound A is regarded as the end of the reaction. The reaction solution was washed with saturated brine (20mL), dried with anhydrous sodium sulfate, and spin-dried. The crude product was purified by medium-pressure reverse phase (80g industrial pre-packed C18 reverse phase column), and purified gradient water/acetonitrile (90/10- 10/90, v/v), time is 1 hour. The pure product was collected and lyophilized to obtain compound C (white solid, 2.63 g, yield 97%). MS: 735.43 (M+H ⁺ )

Compound C (2.63 g, 3.60 mmol), compound D (0.81 g, 3.60 mmol) and DIEA (0.93 g, 7.20 mmol) were dissolved in dichloromethane (20 mL), and the reaction was stirred at room temperature under nitrogen protection for 18 hours. LCMS showed the reaction If the compound C is less than 3% in the solution, the reaction is deemed complete. The reaction solution was washed successively with citric acid aqueous solution (20mL) and saturated brine (20mL), dried over anhydrous sodium sulfate, and spin-dried. The crude product was purified by medium pressure reverse phase (120g industrial pre-packed C18 reverse phase column was selected), and the purification gradient Water/acetonitrile (90/10-10/90, v/v), time 1 hour. The pure product was collected and lyophilized to obtain compound E (white solid, 2.49 g, yield 86%). MS: 807.56 (M+H ⁺ )

Compound E (2.49g, 3.08mmol) and diethylamine (5mL) were added to dichloromethane (20mL), and the reaction was stirred at room temperature under nitrogen protection for 4 hours. LCMS showed that the compound E in the reaction solution was less than 3%, which was regarded as the end of the reaction. . The reaction solution was spin-dried, and the crude product was purified by medium pressure reverse phase (120g industrial pre-packed C18 reverse phase column), and purified gradient water/acetonitrile (90/10-10/90, v/v) for 1 hour. The pure product was collected and lyophilized to obtain compound F (white solid, 1.67 g, yield 93%). MS: 585.36 (M+H ⁺ )

Compound G (405mg, 1.92mmol), HATU (730mg, 1.92mmol) and DIEA (495mg, 3.84mmol) were added to dichloromethane (20mL), the reaction was stirred at room temperature under nitrogen for 30 minutes, and then compound F (1.12g , 1.92 mmol), and the reaction was stirred at room temperature for 4 hours under the protection of nitrogen. LCMS showed that the compound F in the reaction solution was less than 3% as the end of the reaction. The reaction solution was washed with citric acid aqueous solution (20mL), saturated brine (20mL), dried over anhydrous sodium sulfate and spin-dried. The crude product was purified by medium pressure reverse phase (80g industrial pre-packed C18 reverse phase column was selected), and the purification gradient Water/acetonitrile (90/10-10/90, v/v), time 1 hour. The pure product was collected and lyophilized to obtain compound H (white solid, 1.30 g, yield 87%). MS: 778.43 (M+H ⁺ )

MC-MMAF synthetic condensation scheme 1

Compound H (1.25g, 1.60mmol), HATU (641mg, 1.69mmol) and DIEA (437mg, 3.38mmol) were added to dichloromethane (20mL), the reaction was stirred at room temperature under nitrogen for 30 minutes, and then compound I (317mg , 1.92 mmol), and the reaction was stirred at room temperature for 4 hours under nitrogen protection. LCMS showed that the compound H in the reaction solution was less than 3%, which was regarded as the end of the reaction. The reaction solution was washed with citric acid aqueous solution (20mL), saturated brine (20mL), dried over anhydrous sodium sulfate, spin-dried, and the crude product was purified by medium pressure reverse phase (80g industrial pre-packed C18 reverse phase column), purification gradient Water/acetonitrile (90/10-10/90, v/v), time 1 hour. The pure product was collected and lyophilized to obtain compound MC-MMAF (white solid, 1.11 g, yield 75%, HPLC purity 99% by UV 220nm). MS: 925.66 (M+H ⁺ )

MC-MMAF Synthetic Condensation Scheme 2

Compound H (1.12g, 1.43mmol), compound B (198mg, 1.72mmol) and compound EDCI (329mg, 1.72mmol) were dissolved in dichloromethane (10mL), and the reaction was stirred at room temperature under nitrogen protection for 2 hours. LCMS showed the reaction solution Less than 2% of the compound H is regarded as the end of the reaction. The reaction solution was washed with saturated brine (10 mL), dried with anhydrous sodium sulfate, and spin-dried. The crude product was purified by medium-pressure reverse phase (80g industrial pre-packed C18 reverse phase column), and purified gradient water/acetonitrile (90/10- 10/90, v/v), time is 1 hour. The pure product was collected and lyophilized to obtain compound J (white solid, 1.17 g, yield 97%). MS: (M+H ⁺ )

Compound J (1.17g, 1.34mmol), Compound I (0.26g, 1.60mmol) and DIEA (0.35g, 2.70mmol) were dissolved in dichloromethane (10mL), and the reaction was stirred at room temperature for 4 hours under nitrogen protection. LCMS showed the reaction If the compound J is less than 3% in the solution, the reaction is deemed complete. The reaction solution was washed with citric acid aqueous solution (20mL) and saturated brine (20mL) successively, dried over anhydrous sodium sulfate and spin-dried. The crude product was purified by medium pressure reverse phase (80g industrial pre-packed C18 reverse phase column was selected), and the purification gradient Water/acetonitrile (90/10-10/90, v/v), time 1 hour. The pure product was collected and lyophilized to obtain compound MC-MMAF (white solid, 1.15 g, yield 93%). MS: 925.66 (M+H ⁺ )

Claims

An intermediate compound for the synthesis of MC-MMAF, its structural formula is:

Wherein, R is selected from one of hydrogen, succinimidyl, pentafluorophenyl, p-nitrophenyl, and phthalamide.
A method for synthesizing MC-MMAF, characterized in that the method is to make the structural formula
The compound and structural formula are
The compound undergoes condensation reaction in the solvent,

R is hydrogen. During the reaction, under the action of reagent M, reagent N is added. The reagent M is selected from DCC, DCEP, EDC, DIC, HATU, HBTU, HBPIPU, HBPyU, HSPyU, HCTU, HOTU, HOTT, HSTU, HDMA , TATU, TBTU, TCTU, TCFH, TDBTU, TOTU, TOTT, TPTU, TFFH, BTFFH, TNTU, TSTU, COMU, T3P, BOP, PyBOP, PyBrOP, PyClOP, Brop, PyAOP, PyCIU, CDI, TPSI, TSTU, DEPBT One or more of, DMTMM, EEDQ, CIP, CIB, DMC, HOAt, HOBt and EDCI, the reagent N is selected from triethylamine, diisopropylethylamine (DIEA), pyridine, N, N- One or more of dimethyl-4-pyridine;

The solvent is selected from dichloromethane, dimethyl sulfoxide, N,N-dimethylformamide, N,N-dimethylacetamide, tetrahydrofuran, 1,4-dioxane and 2-methyl One or more of tetrahydrofuran;

The reaction temperature is -20°C to 40°C.
The method according to claim 2, wherein the reagent M is a mixture of EDCI and HOBt, and the reagent N is diisopropylethylamine (DIEA).
The method according to claim 2, wherein the reaction temperature is -10°C to 25°C.
A method for synthesizing MC-MMAF, characterized in that, the method is such that the structural formula is
The compound and structural formula are
The compound undergoes condensation reaction in the solvent,

R is selected from one or more of succinimidyl, pentafluorophenyl, p-nitrophenyl, and phthalamide, and reacts under the action of reagent P, which is selected from three One of ethylamine, diisopropylethylamine (DIEA), pyridine, N,N-dimethyl-4-pyridine, sodium carbonate, sodium bicarbonate, potassium carbonate, potassium bicarbonate, lithium carbonate and lithium bicarbonate Kind or more

The solvent is selected from dichloromethane, dimethyl sulfoxide, N,N-dimethylformamide, N,N-dimethylacetamide, tetrahydrofuran, 1,4-dioxane and 2-methyl One or more of tetrahydrofuran;

The reaction temperature is 0°C to 100°C.
The method according to claim 5, wherein the reagent P is sodium carbonate or diisopropylethylamine (DIEA).
The method according to claim 5, wherein the reaction temperature is 15°C-50°C.
The method according to any one of claims 2-7, wherein after the reaction is completed, the MC-MMAF is separated from the reaction liquid.
8. The method according to claim 8, wherein the separation operation comprises evaporating the solvent under reduced pressure, and then purifying or recrystallization by medium pressure chromatography.
A synthesis
Method, characterized in that the compound
HATU and DIEA are stirred at room temperature under nitrogen protection, and then the compound is added
The reaction was carried out by stirring at room temperature under nitrogen protection.