mmaf

MMAF (Monomethylauristatin F) is an antitubulin agent that inhibit cell division; inhibits H3397 cell growth with an IC50 of 105 nM.

(S)-2-((2R, 3R)-3-((S)- 1 -((6S , 9S, 12S, 1 3R)- 12-((S)-sec-butyl)-6, 9-diisopropyl- 13- methoxy-2, 2, 5, 11 -tetramethyl-4, 7, 1 0-trioxo-3 -oxa-5 , 8, 11 -triazapenta-decan- 15- oyl)pyrrolidin-2-yl)-3 -methoxy-2-methylpropanamido)-3 -phenylpropanoic acid (25 mg, 0.030 mmol) in the mixture of HC1 (conc. 0.3 ml) and 1, 4-dioxane (0.9 ml) was stirred at RT for 35 mm. The mixture was diluted with EtOH (1.0 ml) and toluene (1.0 ml), concentrated and re-evaporated with EtOH/toluene (2:1) to afford the title compound as a white solid (22 mg, -400percent yield) for the next step without further purification. LC-MS (ESI) mlz calcd. for C39H66N508 [M+Hj: 732.48, found: 732.60.Compound 12-5 was dissolved in 4N HClDioxane. The reactionmixture was stirred at room temperature for 2 hours and concentrated in vacuo and purified by HPLCto give 150mg of compound 12-6.General procedure: LiOHH2O (5 mg, 0.12 mmol) was added to a solution of 18a (30 mg, 0.06 mmol) in MeOH (2 mL) and water (1 mL) at room temperature. After stirring at room temperature for 3 h, the reaction mixture was concentrated in vacuo and the obtained residue was redissolved in water (5 mL). The mixture was acidified to pH 4e5 by addition of 1 N HCl and a white precipitate was formed. The solid was collected and dried to afford the free acid species as a white solid. The obtained solid was dissolved in CH2Cl2 (2.5 mL), TFA (68 mg, 0.6 mmol) was added and the reaction mixture was stirred at room temperature for about 5 h and then concentrated under reduced pressure. The crude residue was purified by preparative HPLC (SB-C18 column, 5 mm, 9.4 250 mm, 40percent CH3CN/60percent H2O) to yield the desired product (24 mg, 93percent over 2 steps) as a white solid;The compound 62 (57.0 mg, 0.2 mmol), HOSt (27.2 mg, 0.2 mmol), DIC (29.3 IL, 0.2 mmol) and DIPEA (0.2 mmol, 33.1 pL) were added to DMF (6 mE). Afier 30 mm, the compound 63 (109.8 mg, 0.15 mmol) and was added at 0° C. The mixture was allowed to stir at room temperature for 24 hours. The solution was concentrated and purified by prep-HPEC to give compound 64 (94.8 mg, 47.6percent) as a white solid. EC-MS mlz (ES), 1000.53 (M+H).Compound 68 (45.0 mg, 0.14 mmol), HOSt (19.0 mg, 0.14 mmol), DIC (20.5 IL, 0.14 mmol) and DIPEA (23.2 IL, 0.14 mmol, ) were added to DMF (10 mE) and stirred at 0° C. in an ice bath for 1 hout Compound 63 (76.9 mg, 0.11 mmol) was added to the solution. The mixture was allowed to warm to room temperature and lefi to react for 36 hours. The reaction mixture was purified by prep-HPEC to yield compound 69 (64.3 mg, 44.3percent) as a white solid. EC-MS mlz (ESj, 1039.49 (M+H).Step 3. Synthesis of ((2R, 3R)-3-((S)-1-((3R, 4S, 5S)-4-((S)-2-((S)-2-(4-(3-(((5- (azidomethyl)pyrimidin-2-yl)thio)methyl)-1, 1, 3, 3-tetramethyldisiloxaneyl)-N- methylbutanamido)-3-methylbutanamido)-N, 3-dimethylbutanamido)-3-methoxy-5- methylheptanoyl)pyrrolidin-2-yl)-3-methoxy-2-methylpropanoyl)-L-phenylalanine (0216) [00125] A solution of 4-(3-(((5-(azidomethyl)pyrimidin-2-yl)thio)methyl)-1, 1, 3, 3- tetramethyldisiloxanyl)butanoic acid (16.38 mg, 0.041 mmol) and HATU (15.58 mg, 0.041 mmol) in DMF (1997 mul) was stirred for 15 min then charged with a solution of (S)-2- ((2R, 3R)-3-((S)-1-((3R, 4S, 5S)-4-((S)-N, 3-dimethyl-2-((S)-3-methyl-2- (methylamino)butanamido)butanamido)-3-methoxy-5-methylheptanoyl)pyrrolidin-2-yl)-3- methoxy-2-methylpropanamido)-3-phenylpropanoic acid (20 mg, 0.027 mmol) and TEA (6.47 muL, 0.046 mmol) in DMF (1331 muL). After stirring at rt for 7 hrs, the reaction was diluted with DCM (20 mL) and washed with water (2x10 mL). The filtrate was washed with brine, dried over MgSO4, filtered, and concentrated. This material was carried on to the hydrolysis step. MS (ES+): m/z = 1, 113.74 [M+H]+; LCMS: tR = 2.77 min.To a vial containing 4, 4-DIFLUORO-5, 7-DIMETHYL-4-BORA-3A, 4A-DIAZA-S-INDACENE-3- PENTANOIC ACID (93, 3.2 mg , 0.01 mmol) in DCM (0.4 mL) and DMF (0.1 mL) was added DIPEA (1 drop) and HATU (3.9 mg, 0.01 mmol). The mixture was stirred for 0.5 h and transferred to a vial containing Monomethyl Auristatin-F (CAS 74501 7-94-1 ) (7.3 mg, 0.01 mmol). The reaction was stirred at rt overnight, concentrated in vacuo, and the residue purified by reverse phase HPLC (Method B) to give the title compound B81 (5.9 mg , 63percent) as a white solid . LC-MS (Protocol D): m/z 1034.6; Retention time = 7.80 and 8.01 min

Molecular Weight:732.0
XLogP3:2.1
Hydrogen Bond Donor Count:4
Hydrogen Bond Acceptor Count:9
Rotatable Bond Count:21
Exact Mass:731.48331405
Monoisotopic Mass:731.48331405
Topological Polar Surface Area:167
Heavy Atom Count:52
Complexity:1160
Defined Atom Stereocenter Count:9
Covalently-Bonded Unit Count:1
Compound Is Canonicalized:Yes