CN109824759A - A kind of Preparation Method And Their Intermediate of the drug-linker MC-MMAF for antibody drug conjugates - Google Patents
A kind of Preparation Method And Their Intermediate of the drug-linker MC-MMAF for antibody drug conjugates Download PDFInfo
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Abstract
The present invention provides the Preparation Method And Their Intermediate of drug-linker MC-MMAF for antibody coupling drug a kind of.Preparation method of the invention improves the reactivity of N-terminal, to be effectively controlled the generation of racemization；Toxin MMAF is not used directly, and uses the lower compound of toxicity, reduces operation difficulty when mass production；It is prepared without reverse phase, it is easy to operate.
Description
Technical field
The present invention relates to organic synthesis field, in particular to a kind of drug-linker MC- for antibody drug conjugates
The Preparation Method And Their Intermediate of MMAF.
Background technique
Antibody drug conjugates (Antibody drug conjugate, abbreviation ADC) are a kind of novel antineoplastics
Object, principle are that cytotoxin is connected on antibody, and the identification by antibody to cancer cell surfaces specific antigen passes through endocytosis
Effect enters cancer cell and achievees the purpose that magnetic target therapy malignant tumour so that cytotoxin is transported target spot.ADC and biography
The small molecule, anti-tumor drug of system is compared, because can be standby special by the targets identification of antibody and the high activity of toxin, therefore more
Property and validity.
ADC includes three different component parts, i.e. antibody, connexon and cytotoxin.Antibody realizes targeting, connection
Son guarantees the stability in ADC in blood transport process, and after reaching action target spot, toxin plays the killing to cancer cell and makees
With.According to the difference of mechanism of action, the toxin suitable for ADC is divided into micro-pipe class inhibitor (Microtubule
Inhibitors), DNA damage agent (DNA damaging agents), RNA polymerase inhibitor (RNA polymerase
Inhibitors) etc..Currently, being sold in the market with toxin used by the ADC in clinical test is mainly micro-pipe class inhibitor,
It mainly include the compound based on aplysiatoxin (Dolastatin-based) design, such as MMAE, MMAF and MMAD, Yi Jiji
In the compound of maytansine (Maytansine-based) design, such as DM1 and DM4.It is main to be applied for not in terms of connexon
Cleavable type, such as valine-citrulline (Valine-Citriline) and hexahydrobenzoid acid (MCC), after lysosomal hydrolysis,
Drug is still active, and is combined together by bonding pad and some amino acid residue.
There are many generation types of antibody drug conjugates.Both it can be connected by amino on antibody or sulfydryl with drug
Son carries out chemical reaction coupling, can also modify antibody, after introducing specific function base on antibody, then connects with drug
Son carries out chemical reaction coupling or enzymic catalytic reaction coupling.Antibody drug conjugates MC-MMAF structure of the present invention is as follows
It is shown.
The synthetic route of current MC-MMAF reported in the literature is to use toxin MMAF and MC-hex-Acid (1- maleimide
Amido n-caproic acid) dehydration occurs, obtain MC-MMAF.The structure of MMAF are as follows:
Synthetic schemes reported in the literature are as follows:
A methyl is had on the N of the N terminal valine of route MMAF, steric hindrance is larger, in this case, by 1- horse
Come acylimino n-caproic acid be connected on MMAF reaction speed will be relatively slow, even if will also result in using different amide condensed dose
The chiral carbon racemization of the hydrocinnamamide base connection of MMAF.The route is used for the MC-MMAF less than 1g and synthesizes, and finally to apply high pressure
Reverse phase preparation removes Isomeric impurities, and yield is lower than 50%.
The reaction route shows certain defect when amplifying production, such as: 1. this method are because condensing agent can be simultaneously
The carboxyl on MMAF is activated, and causes the racemization of 30-50%, the isomer impurities for being difficult to remove is formed, influences yield；2. due to
Steric hindrance reason above-mentioned, the reaction time is long, and impurity is more, causes difficulty to the post-processing and purifying of reaction；3. final product needs
The preparation of high pressure reverse phase removes isomers, increases operating cost；4. being directly raw material using toxin MMAF, on a large amount of synthetic operations
It need to properly protect, choose safeguard, bring obstacle for production operation.
Summary of the invention
On the one hand, in view of the defects existing in the prior art, the present invention provides the synthetic method of MC-MMAF a kind of, the reaction
Key be to be with structural formulaChemical combination
Object and L-phenylalanineCondensation reaction occurs and directly obtains MC-MMAF or its salt, R is selected from hydrogen, amber
One of imide, pentafluorophenyl group, p-nitrophenyl, phthalyl amido are a variety of.
Above-mentioned purpose of the invention is realized using following technical scheme.
1) synthetic method includes the following steps: compound
It is dissolved in suitable solvent, withAmide condensed reaction occurs, obtains MC-MMAF.
Preferably, in step 1), the suitable solvent is selected from methylene chloride, dimethyl sulfoxide, N, N- dimethyl methyl
One of amide, DMAC N,N' dimethyl acetamide, tetrahydrofuran, 1,4- dioxane and 2- methyltetrahydrofuran are a variety of；More
Preferably, the suitable solvent is selected from one of methylene chloride and n,N-Dimethylformamide or a variety of.
Preferably, in step 1), if R is hydrogen, reagent N is added under the action of reagent M, 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、DMTMM、EEDQ、CIP、CIB、DMC、
One of HOBt and EDCI or a variety of；It is highly preferred that the reagent M in EDCI, EDC, DIC, HOAt and HOBt one
Kind is a variety of；It is further preferred that the reagent M is the mixture of EDCI, EDC or DIC and HOAt or HOBt；Most preferably,
The reagent M is the 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 reaction is -20 DEG C~40 DEG C, preferably -
10 DEG C~25 DEG C.
Preferably, in step 1), if R is amber imide, pentafluorophenyl group, p-nitrophenyl, phthalyl amido
One of or it is a variety of, under the action of reagent P, withIt reacts, obtains MC-MMAF.The reagent P
Selected from triethylamine, diisopropylethylamine (DIEA), pyridine, N, N- dimethyl -4- pyridine, sodium carbonate, sodium bicarbonate, potassium carbonate,
One of saleratus, lithium carbonate and lithium bicarbonate are a variety of, preferably sodium carbonate or diisopropylethylamine (DIEA).Instead
Answering temperature is 0 DEG C~100 DEG C, preferably 15 DEG C~50 DEG C.
It preferably, further include after completion of the reaction, the step of MC-MMAF is separated from reaction solution in step 1).
Preferably, the separation includes chromatogram purification or recrystallization being pressed in use, then to obtain by evaporated under reduced pressure solvent
MC-MMAF。
Preparation method of the invention improves the reactivity of N-terminal, to be effectively controlled the generation of racemization；No
Toxin MMAF is directly used, and uses the lower L-phenylalanine of toxicity, reduces operation difficulty when mass production；Without anti-
Mutually prepare, it is easy to operate.As described above, so that quality standard is relatively easy to control, preparing hectogram grade this method reduce operation difficulty
It can not apply.
On the other hand, this patent also provides the midbody compound of synthesis MC-MMAF, and structural formula isWherein, R is selected from
One of hydrogen, succinimido, pentafluorophenyl group, p-nitrophenyl, phthalyl amido.Preferably following compound,
It is as shown in table 1:
Table 1
In another aspect, the present invention also provides a kind of synthesis
Method, synthesis step is as follows,
Comprising synthesis important intermediate H in this route, the not no report of synthetic method before the compound.
Compound H is synthesized, usual thinking is with following route:
However experiment discovery H1 can be decomposed during de- tert-butyl, yield is very low.
And if not protecting carboxyl, in condensation course, the carboxyl of compound F can also participate in reacting, and cause to react very complicated,
There are many by-product.
Reaction 30 minutes will be first stirred at room temperature in the method that the present invention uses under compound G, HATU and DIEA nitrogen protection, then
Addition compound F is stirred at room temperature under nitrogen protection carries out reaction 4 hours.Charging sequence and feed ratio are improved by making repeated attempts
Rate makes the carboxyl of compound F be not involved in reaction, can obtain compound H with very high yield.By changing reaction condition, greatly
The yield for improving compound H greatly makes the route have the possibility applied to production.
The present invention abandons existing MMAF synthetic route, and MC-MMAF is regarded as an entirety to synthesize.Maximum is asked
Topic is that MC connexon is the higher segment of reactivity ratio, and MC being connected can make to synthesize difficulty in advance increases, art technology
Personnel not will recognize that this route.We solve by much studying and are introduced into MC fragment compound shakiness in synthesis in advance
Fixed problem can be realized this whole synthetic route.
As used herein, the definition of common organic matter abbreviation and its it is corresponding No. CAS as shown in table 2:
Table 2
Detailed description of the invention
Fig. 1 is the high-efficient liquid phase chromatogram for the DMT-3 that the present invention synthesizes.
Fig. 2 is the liquid chromatogram for the compound A that the present invention synthesizes.
Fig. 3 is the mass spectrogram for the compound A that the present invention synthesizes.
Fig. 4 is the liquid chromatogram for the compound C that the present invention synthesizes.
Fig. 5 is the mass spectrogram for the compound C that the present invention synthesizes.
Fig. 6 is the liquid chromatogram for the compound E that the present invention synthesizes.
Fig. 7 is the mass spectrogram for the compound E that the present invention synthesizes.
Fig. 8 is the liquid chromatogram for the compound F that the present invention synthesizes.
Fig. 9 is the mass spectrogram for the compound F that the present invention synthesizes.
Figure 10 is the liquid chromatogram for the compound H that the present invention synthesizes.
Figure 11 is the mass spectrogram for the compound H that the present invention synthesizes.
Figure 12 is the liquid chromatogram for the target product MC-MMAF that condensation scheme 1 of the present invention synthesizes.
Figure 13 is the mass spectrogram for the target product MC-MMAF that condensation scheme 1 of the present invention synthesizes.
Figure 14 is the nmr spectrum for the target product MC-MMAF that condensation scheme 1 of the present invention synthesizes.
Figure 15 is the high-efficient liquid phase chromatogram for the target product MC-MMAF that condensation scheme 1 of the present invention synthesizes
Figure 16 is the liquid chromatogram for the compound J that the present invention synthesizes.
Figure 17 is the mass spectrogram for the compound J that the present invention synthesizes.
Figure 18 is the liquid chromatogram for the target product MC-MMAF that condensation scheme 1 of the present invention synthesizes.
Figure 19 is the mass spectrogram for the target product MC-MMAF that condensation scheme 1 of the present invention synthesizes.
Specific embodiment
Technical solution of the present invention is further non-limitingly described in detail With reference to embodiment.It needs
, it is noted that the technical concepts and features of following embodiments only to illustrate the invention, its object is to allow be familiar with technique
Personage cans understand the content of the present invention and implement it accordingly, and it is not intended to limit the scope of the present invention.It is all according to this hair
Equivalent change or modification made by bright Spirit Essence, should be covered by the protection scope of the present invention.
LCMS indicates LC-MS detection method；HPLC indicates high performance liquid chromatography detection.
The raw materials and reagents of every single step reaction according to the present invention are commercially available or according to the method for the invention system
?.
The present invention provides a kind of method for synthesizing MC-MMAF, which includes the following steps:
1) by compoundIt is suitable to be dissolved in
Solvent in, the suitable solvent be selected from methylene chloride, dimethyl sulfoxide, n,N-Dimethylformamide, N, N- dimethyl second
One of amide, tetrahydrofuran, Isosorbide-5-Nitrae-dioxane and 2- methyltetrahydrofuran are a variety of, withHair
Raw amide condensed reaction, obtains MC-MMAF.
In step 1), if R be hydrogen, then under the action of reagent M, be added reagent N, the reagent M be 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
One of or it is a variety of；It is highly preferred that the reagent M is selected from one of EDCI, EDC, DIC, HOAt and HOBt or a variety of；Into
Preferably, the reagent M is the mixture of EDCI, EDC or DIC and HOAt or HOBt to one step；Most preferably, the reagent M is
The mixture of EDCI and HOBt.The reagent N is selected from triethylamine, diisopropylethylamine (DIEA), pyridine, N, N- dimethyl -4-
Pyridine, preferably diisopropylethylamine (DIEA).Reaction temperature is -20 DEG C~40 DEG C, preferably -10 DEG C~25 DEG C.
In step 1), if R is one of amber imide, pentafluorophenyl group, p-nitrophenyl, phthalyl amido
Or it is a variety of, then under the action of reagent P, withIt reacts, obtains MC-MMAF.The reagent P is selected from
Triethylamine, diisopropylethylamine (DIEA), pyridine, N, N- dimethyl -4- pyridine, sodium carbonate, sodium bicarbonate, potassium carbonate, carbonic acid
One of hydrogen potassium, lithium carbonate and lithium bicarbonate are a variety of, preferably sodium carbonate or diisopropylethylamine (DIEA).Reaction temperature
Degree is 0 DEG C~100 DEG C, preferably 15 DEG C~50 DEG C.
It further include after completion of the reaction, the step of MC-MMAF is separated from reaction solution, it is preferable that institute in step 1)
Stating separation includes chromatogram purification or recrystallization being pressed in use, then to obtain MC-MMAF by evaporated under reduced pressure solvent.
The present invention also provides a kind of synthesisMethod,
First by compoundReaction is stirred at room temperature under HATU and DIEA nitrogen protection, adds compoundIt is stirred at room temperature and is reacted under nitrogen protection.
Embodiment 1
The reaction route of the present embodiment is as follows:
Condensation scheme 1
Condensation scheme 2
In 3L there-necked flask, 1.5L methylene chloride and Dil.HCl (202.3g, 0.683mol 1.0eq) is added, magnetic force stirs
Mix, nitrogen protection, sequentially add Z-Val-OH (163.23g, 0.65mol, 0.95eq) and HATU (311.6g, 0.82mol,
1.20eq), it is cooled to ice bath after 30min clock is stirred at room temperature, 10 degree of dropwise addition DIEA (452.5ml, 4.0eq) of temperature control are added dropwise,
After stirring 30min under ice bath, room temperature is moved to, reacts 16h, HPLC detection, main peak is product peak (retention time 29.98min), former
Expect that Dil.HCl fully reacting, reaction terminate.Reaction solution washs (2L*1) with aqueous citric acid solution, and saturated sodium bicarbonate solution is washed
It washs (2L*1), saturated common salt water washing (2L*1), after organic layer is with anhydrous sodium sulfate drying, filters, precipitation obtains crude product 531g. should
Crude product is dissolved in 800ml methanol, and 1.1ml (1mol/L) dilute hydrochloric acid (about 1 hour) is added dropwise under ice bath stirring, 12h is stirred at room temperature, stops
Stirring, layering separate upper aqueous layer, and lower layer's product is drawn with oil pump and done, and obtain 325g DMT-1, yield 91%
In 2L single port bottle, 800ml methanol and DMT-1 (LN114-38,325g, 0.66mol) and 110gPd (OH) is added
2/C, H2 are replaced three times, react at room temperature 5h, and TLC monitors raw material DMT-1 fully reacting.Diatomite is added in sand core funnel, filters,
And filter cake is washed with 1L methanol, filtrate is collected, filtrate is evaporated, and it is not blistering that oil pump is evacuated to product, obtains DMT-2 230.2g, purity
94%；Yield: 97%.
In 3L there-necked flask, DMT-2 (LN114-40-01,230.2g, practical 0.60mol, 1.0eq) is dissolved in
In 500mlDCM, be sufficiently stirred, be added Fmoc-Me-val (202.6g, 0.57mol, 0.95eq) and HATU (292.9g,
0.77mol, 1.20eq), 1L DCM is added, is cooled to ice bath, 10 degree of dropwise addition DIEA of temperature control after 30min is stirred at room temperature
(212.7ml, 2.0eq), is added dropwise, and after stirring 30min under ice bath, moves to room temperature, reacts 16.0h, HPLC detection, and main peak is
Product peak (retention time 36.00min), raw material DMT-2 fully reacting, reaction terminate.Reaction solution with water washing (2.0L*1),
Aqueous citric acid solution washs (2L*1), and saturated sodium bicarbonate solution washs (2L*1), saturated common salt water washing (1L*1), organic layer
With anhydrous sodium sulfate it is dry after, filter, precipitation obtains the crude product 655g. crude product and is dissolved in 650ml methanol, stirs and lower 360ml is added dropwise
12h is stirred at room temperature in (1mol/L) dilute hydrochloric acid, stops stirring, is layered, separation upper aqueous layer, and such twice.Lower layer's product is with oil pump
It draws and does, obtain 373g DMT-3, HPLC purity is 96.7%, yield 90%.
Compound DMT-3 (173g, 0.25mol) addition methylene chloride (500mL) is sufficiently stirred, is added three under ice bath
Fluoroacetic acid (170mL) is slowly increased to room temperature under nitrogen protection, is stirred to react 16 hours, and LCMS shows compound in reaction solution
DMT-3 is considered as reaction less than 1% and terminates.Reaction solution is washed with water (500mL × 3), then is washed with saturated salt solution (500mL)
It washs, organic phase anhydrous Na2SO4Dry, Rotary Evaporators are concentrated to give faint yellow crude product 165g；1.1L (petroleum is added in crude product
Ether: ethyl acetate=10:1) in the mixed solvent mechanical stirring, white solid can be precipitated after about 1 hour, filter, filter cake DCM
(500mL) is washed twice, and vacuum drying obtains compound A (136g, yield 85%).MS:638.27 (M+H+)
By compound A (2.38g, 3.71mmol), compound B (456mg, 3.96mmol) and compound EDCI (760mg,
It 3.96mmol) is dissolved in methylene chloride (20mL), reaction 2 hours is stirred at room temperature under nitrogen protection, LCMS shows chemical combination in reaction solution
Object A is considered as reaction less than 5% and terminates.Reaction solution is washed with saturated salt solution (20mL), and anhydrous sodium sulfate is dry, is spin-dried for, crude product
Purified (the pre-filled C18 reversed-phase column of industry for selecting 80g) with middle pressure reverse phase, purifies gradient water/acetonitrile (90/10-10/90, v/
V), time 1 hour.Pure product freeze-drying is collected, compound C (white solid, 2.63g, yield 97%) is obtained.MS:735.43
(M+H+)
By compound C (2.63g, 3.60mmol), compound D (0.81g, 3.60mmol) and DIEA (0.93g,
It 7.20mmol) is dissolved in methylene chloride (20mL), reaction 18 hours is stirred at room temperature under nitrogen protection, LCMS shows chemical combination in reaction solution
Object C is considered as reaction less than 3% and terminates.Reaction solution is successively washed with aqueous citric acid solution (20mL) and saturated salt solution (20mL),
Anhydrous sodium sulfate is dry, is spin-dried for, and presses reverse phase to purify (the pre-filled C18 reversed-phase column of industry for selecting 120g) in crude product, purifying ladder
It spends water/acetonitrile (90/10-10/90, v/v), the time 1 hour.Collect pure product freeze-drying, obtain compound E (white solid,
2.49g, yield 86%).MS:807.56 (M+H+)
Compound E (2.49g, 3.08mmol) and diethylamine (5mL) are added to methylene chloride (20mL), under nitrogen protection
Reaction 4 hours is stirred at room temperature, compound E, which is considered as reaction less than 3%, in LCMS display reaction solution terminates.Reaction solution is spin-dried for, crude product
Purified (the pre-filled C18 reversed-phase column of industry for selecting 120g) with middle pressure reverse phase, purifies gradient water/acetonitrile (90/10-10/90, v/
V), time 1 hour.Pure product freeze-drying is collected, compound F (white solid, 1.67g, yield 93%) is obtained.MS:585.36
(M+H+)
By compound G (405mg, 1.92mmol), HATU (730mg, 1.92mmol) and DIEA (495mg, 3.84mmol)
Be added to methylene chloride (20mL), be stirred at room temperature under nitrogen protection reaction 30 minutes, add compound F (1.12g,
1.92mmol), reaction 4 hours is stirred at room temperature under nitrogen protection, compound F is considered as reaction less than 3% in LCMS display reaction solution
Terminate.Reaction solution is successively used aqueous citric acid solution (20mL), and saturated salt solution (20mL) washing, anhydrous sodium sulfate is dry, is spin-dried for,
Reverse phase is pressed to purify (the pre-filled C18 reversed-phase column of industry for selecting 80g) in crude product, purifying gradient water/acetonitrile (90/10-10/90,
), v/v time 1 hour.Pure product freeze-drying is collected, compound H (white solid, 1.30g, yield 87%) is obtained.MS:
778.43(M+H+)
MC-MMAF synthetic condensation agent scheme 1
By compound H (1.25g, 1.60mmol), HATU (641mg, 1.69mmol) and DIEA (437mg, 3.38mmol)
Be added to methylene chloride (20mL), be stirred at room temperature under nitrogen protection reaction 30 minutes, add compound I (317mg,
1.92mmol), reaction 4 hours is stirred at room temperature under nitrogen protection, compound H is considered as reaction less than 3% in LCMS display reaction solution
Terminate.Reaction solution is successively used aqueous citric acid solution (20mL), and saturated salt solution (20mL) washing, anhydrous sodium sulfate is dry, is spin-dried for,
Reverse phase is pressed to purify (the pre-filled C18 reversed-phase column of industry for selecting 80g) in crude product, purifying gradient water/acetonitrile (90/10-10/90,
), v/v time 1 hour.Collect pure product freeze-drying, obtain compound MC-MMAF (white solid, 1.11g, yield 75%,
HPLC purity 99%by UV 220nm).MS:925.66 (M+H+)
MC-MMAF synthetic condensation agent scheme 2
By compound H (1.12g, 1.43mmol), compound B (198mg, 1.72mmol) and compound EDCI (329mg,
It 1.72mmol) is dissolved in methylene chloride (10mL), reaction 2 hours is stirred at room temperature under nitrogen protection, LCMS shows chemical combination in reaction solution
Object H is considered as reaction less than 2% and terminates.Reaction solution is washed with saturated salt solution (10mL), and anhydrous sodium sulfate is dry, is spin-dried for, crude product
Purified (the pre-filled C18 reversed-phase column of industry for selecting 80g) with middle pressure reverse phase, purifies gradient water/acetonitrile (90/10-10/90, v/
V), time 1 hour.Pure product freeze-drying is collected, compound J (white solid, 1.17g, yield 97%) is obtained.MS:(M+H+)
By compound J (1.17g, 1.34mmol), compound I (0.26g, 1.60mmol) and DIEA (0.35g,
It 2.70mmol) is dissolved in methylene chloride (10mL), reaction 4 hours is stirred at room temperature under nitrogen protection, LCMS shows chemical combination in reaction solution
Object J is considered as reaction less than 3% and terminates.Reaction solution is successively washed with aqueous citric acid solution (20mL) and saturated salt solution (20mL),
Anhydrous sodium sulfate is dry, is spin-dried for, and presses reverse phase to purify (the pre-filled C18 reversed-phase column of industry for selecting 80g) in crude product, purifies gradient
Water/acetonitrile (90/10-10/90, v/v), the time 1 hour.Pure product freeze-drying is collected, obtaining compound MC-MMAF, (white is solid
Body, 1.15g, yield 93%).MS:925.66 (M+H+)。
Claims (10)
1. a kind of midbody compound for synthesizing MC-MMAF, structural formula are as follows:
Wherein, R is selected from one of hydrogen, succinimido, pentafluorophenyl group, p-nitrophenyl, phthalyl amido.
2. a kind of method for synthesizing MC-MMAF, which is characterized in that the method is to make structural formulaCompound and structure
Formula isCompound condensation reaction occurs in a solvent,
R is hydrogen, when reaction under the action of reagent M, is added reagent N, the reagent M be 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、
One in PyAOP, PyCIU, CDI, TPSI, TSTU, DEPBT, DMTMM, EEDQ, CIP, CIB, DMC, HOAt, HOBt and EDCI
Kind or it is a variety of, the reagent N is selected from triethylamine, diisopropylethylamine (DIEA), pyridine, N, one in N- dimethyl -4- pyridine
Kind is a variety of；
The solvent is selected from methylene chloride, dimethyl sulfoxide, N,N-dimethylformamide, DMAC N,N' dimethyl acetamide, tetrahydro furan
It mutters, one of 1,4- dioxane and 2- methyltetrahydrofuran or a variety of；
Reaction temperature is -20 DEG C~40 DEG C.
3. according to the method described in claim 2, it is characterized in that, the reagent M is the mixture of EDCI and HOBt, the examination
Agent N is diisopropylethylamine (DIEA).
4. according to the method described in claim 2, it is characterized in that, reaction temperature is -10 DEG C~25 DEG C.
5. a kind of method for synthesizing MC-MMAF, which is characterized in that the method is to make structural formulaCompound and structure
Formula isCompound condensation reaction occurs in a solvent,
R is selected from one of succinimido, pentafluorophenyl group, p-nitrophenyl, phthalyl amido or a variety of, in reagent
It reacts under the action of P, the reagent P is selected from triethylamine, diisopropylethylamine (DIEA), pyridine, N, N- dimethyl -4- pyrrole
One of pyridine, sodium carbonate, sodium bicarbonate, potassium carbonate, saleratus, lithium carbonate and lithium bicarbonate are a variety of；
The solvent is selected from methylene chloride, dimethyl sulfoxide, N,N-dimethylformamide, DMAC N,N' dimethyl acetamide, tetrahydro furan
It mutters, one of 1,4- dioxane and 2- methyltetrahydrofuran or a variety of；
Reaction temperature is 0 DEG C~100 DEG C.
6. according to the method described in claim 5, it is characterized in that, reagent P is sodium carbonate or diisopropylethylamine (DIEA).
7. according to the method described in claim 5, it is characterized in that, reaction temperature is 15 DEG C~50 DEG C.
8. the method according to claim 2, which is characterized in that it is described after the reaction was completed, by MC-MMAF from anti-
It answers in liquid and separates.
9. according to the method described in claim 8, it is characterized in that, the lock out operation includes by evaporated under reduced pressure solvent, so
Chromatogram purification or recrystallization are pressed in using afterwards.
10. a kind of synthesisSide
Method, which is characterized in that first by compoundIt is stirred at room temperature under HATU and DIEA nitrogen protection anti-
It answers, adds compoundBe stirred at room temperature under nitrogen protection into
Row reaction.
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| PCT/CN2019/092948 WO2020181686A1 (en) | 2019-03-08 | 2019-06-26 | Preparation method for drug-linker mc-mmaf used for antibody drug conjugates and intermediate thereof |
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| WO2020181686A1 (en) * | 2019-03-08 | 2020-09-17 | 联宁（苏州）生物制药有限公司 | Preparation method for drug-linker mc-mmaf used for antibody drug conjugates and intermediate thereof |
| WO2021056754A1 (en) | 2019-09-29 | 2021-04-01 | 烟台迈百瑞国际生物医药股份有限公司 | Method for preparing antibody-drug conjugate intermediate by means of acid method and application thereof |
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