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CAS No. : | 159857-81-5 | MDL No. : | MFCD22200431 |
Formula : | C35H43N7O11 | Boiling Point : | - |
Linear Structure Formula : | - | InChI Key : | HYSPJPGXSALJRR-DHIFEGFHSA-N |
M.W : | 737.76 | Pubchem ID : | 57587849 |
Synonyms : |
Maleimidocaproyl-L-valine-L-citrulline-p-aminobenzyl alcohol p-nitrophenyl carbonate;MC-VC-PAB-PNP
|
Chemical Name : | 4-((S)-2-((S)-2-(6-(2,5-Dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanamido)-3-methylbutanamido)-5-ureidopentanamido)benzyl (4-nitrophenyl) carbonate |
Signal Word: | Warning | Class: | N/A |
Precautionary Statements: | P261-P305+P351+P338 | UN#: | N/A |
Hazard Statements: | H302-H315-H319-H335 | Packing Group: | N/A |
GHS Pictogram: |
* All experimental methods are cited from the reference, please refer to the original source for details. We do not guarantee the accuracy of the content in the reference.
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
71% | With pyridine; benzotriazol-1-ol; In N,N-dimethyl-formamide; at 20℃; for 24h; | Example 29 Synthesis of N-alkyl maleimido-Val-Cit-PAB-MMAE mc-ValCit-PAB-OH PNP Carbonate "N-alkyl maleimide Val-Cit-PAB-MMAE" Maleimidocaproyl-valine-citrulline-/7-aminobenzylcarbonyl-mono- methyl-Auristatin-E (mc- ValCit-PABC-MMAE) was prepared according to the literature method with modifications.1 Thus, a mixture of mc-Val-Cit-p-aminobenzyl alcohol p-nitrophenylcarbonate (8) freshly prepared according to literature method2 (265 mg, 0.36 mmol, 1.5 eq.), MMAE (9) (169 mg, 0.24 mmol, 1 eq.) and N-hydroxybenzotriazole (HOBt) (6.48 mg, 0.048 mmol, 0.2 eq.) were stirred in DMF (5 mL) at r.t. for 2 min., followed by addition of pyridine (38 mg, 0.48 mmol, 2 eq.). After stirring for 24 h, volatile organics were removed under vacuum. The residue obtained was triturated with ethyl acetate and methanol (1L) to afford mc-Val-Cit-PAB- MMAE (10) as a white powder (220 mg, 0.17 mmol, 71 % yield) that was usually >95% pure by RP-HPLC analysis. If necessary, further purification can be carried out by C18 reversed- phase preparative HPLC or size exclusive column chromatography. Electrospray (ES)-MS m/z 1339 (M+Na, base peak), 1317 (M + 1). |
65% | With 2,6-dimethylpyridine; benzotriazol-1-ol; In N,N-dimethyl acetamide; at 40℃; | A compound of the following formula (1 eq.): and mc-Val-Cit-PABC-PNP (1.18 eq.) were solubilized in N,N-dimethylacetamide (7.87 vol.). l-Hydroxybenzotriazole (HOBt) hydrate (8.95 wt%) and 2,6-lutidine (2.315 vol.) were then added and the reaction mixture was stirred at 40C for 12-16 hours until reaction was complete. The reaction mixture was cooled to 20C and added into tert-Butyl methyl ether (168 vol.). The resultant slurry was stirred for 3-5 hours and filtered, washed and dried under vacuum. Crude product was purified by column purification and product-containing fractions were concentrated to dryness and slurried in Ethyl acetate (20 vol.) before being isolated by filtration, washed and dried (65% yield). MS: m/e 1317 (MH)+, 1339 (M+Na)+. |
Ca. 42% | 20 mg mc-vc-PAB-PNP (1.5 eq) and 3 mg HOBT were added to 2 ml DMF.After stirring for a while at room temperature,Add 13mg MMAE,0.5ml pyridine,25ulDIEA.The reaction was stirred at room temperature for 2 days.After the reaction is completed,The reaction solution is directly purified using a preparative column.Collect the required ingredients and lyophilize them after concentrationGet about 10mg product,The yield is about 42%. |
Ca. 42% | With benzotriazol-1-ol; N-ethyl-N,N-diisopropylamine; In N,N-dimethyl-formamide; at 20℃; for 48h; | 20mg of mc-vc-PAB-PNP (1.5 eq) and 3 mg of HOBT were added to 2 ml of DMF. After stirring for a while at room temperature,Add 13 mg MMAE, 0.5 ml pyridine,25ulDIEA. The reaction solution was stirred at room temperature for 2d.. After the reaction is completed, the reaction solution is directly purified by a preparative column.The desired components are collected and concentrated and lyophilized to obtain about 10 mg of the product.The yield is about 42%. |
With pyridine; benzotriazol-1-ol; In N,N-dimethyl-formamide; for 24h; | Preparation Method of Compound 20-10: MMAE (100.5 mg, 0.14 mmol, 1 eq. ) , Compound 20-9 (110.6 mg, 0.15 mmol, 1.1 eq. ) and HPBt (19 mg, 0.14 mmol, 1.0eq. ) were diluted with DMF (2 mL) . After 2 minutes, pyridine (0.5 mL) was added and the reaction was monitored by reverse-phase HPLC. The reaction was complete after about 24 hours. The mixture was concentrated and the resulting residue was purified using reverse phase preparative-HPLC (Varian Dynamax column 21.4 mm×25 cm, 5mu,through gradient elution of MeCN and Et3N-CO2 (pH 7) at 20 mL/minutes from 10 to 100 for 40 minutes. The relevant fractions were pooled and concentrated to give an off-white solid intermediate, i.e., Compound 20-10. MS [M] +: 1, 315.78. | |
10.7 g | MMAE (7.18g. lOmmol) was measured and added into a 250m1 three-neck flask, dissolved in anhydrous DMF, and stirred till clear at room temperature under N2 protection. Mc-Val-Cit-PAB-PNP (7.37g) and HOAt (72mg, 2mmol) were added into the solution and reacted for 5 mm, and then DIEA (3.5m1, 2Ommol) was added drop-wise, the reaction was continued for 30 mm at roomtemperature, and then the temperature was increased to 40?SOC and reacted for 20h, during which HPLC was used to monitor the reaction. DMF was removed by drying in vacuo, and the product Mc-Val-Cit-PAB-MMAE (10.7g. purity: 99.3%) was obtained by further HPLC purification. | |
With pyridine; benzotriazol-1-ol; In N,N-dimethyl-formamide; at 20℃; | MC-VC-PAB-PNP (93 mg, 0.125 mmol) and MMAE (60 mg, 0.083 mmol) were dissolved in 4 mL DMF.Add HoBt (10 mg, 0.074 mmol) and 0.5 mL of pyridine in sequence.The reaction was stirred at room temperature overnight.The reaction product is isolated and purified using a preparative liquid phase.The chromatographic conditions were as follows: phase A H2O + 0.08% TFA, phase B acetonitrile + 0.08% TFA. 0-35min B phase 65% isocratic elution,The pure product was collected and the reaction product was lyophilized. |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
With pyridine; benzotriazol-1-ol; In N,N-dimethyl-formamide; for 24h; | The activated linker 39 (50 mg, 0.068 mmol), MMAE (32.6 mg, 0.045 mmol) and N-hydroxybenzotriazole (1.4 mg, 0.0091 mmol) are stirred in dry DMF (1 ml) for 2 min. after which a drop of pyridine is added and the reaction stirred for 24 h. The solvent is then removed by high vacuum and the residue purified by reverse-phase preparative HPLC togive the desired product 40 after lyopholisation as a white powder; MS (m/z) 1316.7 (M+H). |
Yield | Reaction Conditions | Operation in experiment |
---|---|---|
200 mg | With 2,6-dimethylpyridine; 2-hydroxy-pyridine N-oxide; In N,N-dimethyl-formamide; for 48h;Inert atmosphere; Sealed tube; | To a scintillation vial were added N-Methyl-L-valine (6.2 equiv.) and mc-Val- Cit-PABC-PNP (500 mg, 1.0 equiv.). The vial was purged with nitrogen (x 3), and the solids were suspended in 2,6-lutidine (4.0 vol.) and DMF (4.0 vol.). Solid HOPO (1.2 equiv.) was added in one portion, the vessel was sealed, and the reaction was stirred vigorously for 48 h. The reaction was poured into MTBE (200 vol.), and the resulting mixture was vacuum filtered (washing with MTBE) to deliver a grey solid. The solid was solubilized in minimal AcOH (4.0 vol.), and the resulting solution was chromatographically purified on silica gel (5% MeOH in CH2Cl2 to 20%) to provide mc-Val-Cit-PAB-<strong>[2480-23-1]N-Me-Val-OH</strong> as a pale-yellow residue (200 mg). |