Monomethylauristatin E

Monomethyl auristatin E (MMAE) is a synthetic derivative of dolastatin 10 and functions as a potent mitotic inhibitor by inhibiting tubulin polymerization. MMAE is widely used as a cytotoxic component of antibody-drug conjugates (ADCs) to treat several different cancer types.

Monomethyl Auristatin E is a dolastatin-10 peptide derivative with potent antimitotic activity and potential antineoplastic activity as part of an antibody-drug conjugate (ADC). Monomethyl auristatin E (MMAE) binds to tubulin, blocks tubulin polymerization, and inhibits microtubule formation, which results in both disruption of mitotic spindle assembly and arrest of tumor cells in the M phase of the cell cycle. To minimize toxicity and maximize efficacy, MMAE is conjugated, via a cleavable peptide linker, to a monoclonal antibody that specifically targets a patient's tumor. The linker is stable in the extracellular milieu but is readily cleaved to release MMAE following binding and internalization of the ADC by the target cells.

0.30 g of stearic acid-sn-2-hydroxyethyl dithiopropionic acid glycerylphosphocholine, 0.10 g of triphosgene (BTC) was uniformly mixed in 100 mL of anhydrous dichloromethane. Under a nitrogen atmosphere, 0.15 g of a DMAP solution in dichloromethane was slowly added dropwise, and the reaction was stirred at room temperature for 2 h. To the above system, 0.1 g of compound MMAE (Monomethyl auristatin E) was added and reacted for 3 hours. After that, the crude product was washed with aq. The product was purified by column chromatography to give the product stearic acid-sn-2-MMAE urethane ethyl dithiopropionate glycerol phosphate choline 0.13 g as a white solid.To the solution of S3 (1 eq., 37.9 mg, 0.055 mmol) and MMAE (1 eq., 39.5 mg, 0.055 mmol) in DMF (1.05 mL) was added a solution of HOBt (1 eq., 7.43 mg, 0.055 mmol) in pyridine (0.263 mL). The mixture was stirred at room temperature for 24h then diluted with MeOH (10.5 mL) and cooled to 0C. To the resulting solution was added LiOH (10 eq., 1 M in H20, 0.55 mL, 0.55 mmol) and the mixture was incubated for at 4C for l6h. The resulting solution was quenched with HCOOH (20 eq., 1 M in H20, 1.1 mL, 1.1 mmol) and concentrated to 3 mL under reduced pressure. The residue was purified by preparative HPLC (method 1) to yield S4 (36.6 mg, 0.0325 mmol, 59 %) as a pale yellow solid. (0188) HPLC (Method 2) RT: 7.66 min (0189) MS (Method 3) m/z : 1127.9 [M+H]+To the solution of R3 (1 eq., 41.3 mg, 0.06 mmol) and MMAE (1 eq., 43.1 mg, 0.06 mmol) in DMF (1.15 mL) was added a solution of HOBt (1 eq., 8.11 mg, 0.06 mmol) in pyridine (0.287 mL). The mixture was stirred at room temperature for 24h then diluted with MeOH (11.5 mL) and cooled to 0C. To the resulting solution was added LiOH (10 eq., 1 M in H20, 0.6 mL, 0.6 mmol) and the mixture was incubated for at 4C for l6h. The resulting solution was quenched with HCOOH (20 eq., 1 M in H20, 1.2 mL, 1.2 mmol) and concentrated to 3 mL under reduced pressure. The residue was purified by preparative HPLC (method 1) to yield R4 (41.9 mg, 0.0372 mmol, 62 %) as a pale yellow solid.HPLC (Method 2) RT: 7.81 minMS (Method 3) m/z : 1127.7 [M+H]+To a 100 ml RB flask containing tert-butyl((S)-3-methyl-1-(((S)-1- ((4-((((4nitrophenoxy)carbonyl)oxy)methyl)phenyl)amino)-1 -oxopropan-2- yl)amino)-1-oxobutan-2-yl)carbamate 4-3 (349 mg, 0.625 mmol), was added A 100 ml RB flask containing terf-butyl((S)-3-methyl-1-(((S)-1-((4- ((((4nitrophenoxy)carbonyl)oxy)methyl)phenyl)amino)-1-oxopropan-2- yl)amino)-1-oxobutan-2-yl)carbamate 7-3 (349 mg, 0.625 mmol) was charged with In order to conjugate MMAE to the peptide-linker conjugate of Chemical Formula 3, the peptide-linker conjugate of Chemical Formula 3 precipitate was dried to obtain a powder (653 mg) and then mixed with MMAE (478 mg, 1.2 eq) and HOBt (56 mg, 0.75 eq) in anhydrous DMF (40 mL). Then, a reaction mixture was obtained by adding pyridine (10 mL) and DIPEA (193 muL, 2 eq) and stirring at room temperature for 72 hours. Then, diethyl ether was poured to synthesize a peptide-linker-MMAE conjugate represented by Chemical Formula 5 as a precipitate.

Dolastatin 10 is a natural antimitotic and antineoplastic agent that binds to tubulin and inhibits tubulin polymerization. Monomethyl Auristatin E (MMAE) is a synthetic analog of dolastatin 10 that similarly inhibits tubulin polymerization and exhibits potent cytotoxicity. It is commonly conjugated with monoclonal antibodies directed at antigens specific to cancer cells for tumor-directed cytotoxicity. MMAE is typically coupled to the antibody via a protease-cleavable linker, allowing separation of the drug from the antibody following intracellular localization.[Cayman Chemical]

Molecular Weight:718.0
XLogP3:4.1
Hydrogen Bond Donor Count:4
Hydrogen Bond Acceptor Count:8
Rotatable Bond Count:20
Exact Mass:717.50404949
Monoisotopic Mass:717.50404949
Topological Polar Surface Area:150
Heavy Atom Count:51
Complexity:1100
Defined Atom Stereocenter Count:10
Covalently-Bonded Unit Count:1
Compound Is Canonicalized:Yes