Proteases screening for the kinetic resolution of amines with N-acyl α-amino acid trifluoromethyl esters: automated docking approach of binding energies using Subtilisin Novo as a prototype for serine proteases

The screening of a panel of 17 proteases resulted in the selection of 4 serine proteases for the resolution of 3-amino-1-phenylbutane. The latter were used to determine the best acyl donor from a series of N-acyl α-amino acid trifluoroethyl esters selected as peptide mimetics (E factor up to 99). The results were correlated to an automated docking determination of their binding affinities for Subtilisin Novo.

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N-Octanoyl l-phenylalanineC17H25NO3Ee >99.5%[α]D25=+75.0 (c 1.06, CHCl3)Source of chirality: l-phenylalanineAbsolute configuration (S)

N-Octanoyl l-phenylalanine trifluoroethyl esterC19H26NO3F3Ee >99.5%[α]D25=+25.5 (c 1.00, CHCl3)Source of chirality: l-phenylalanineAbsolute configuration (S)

N-Acetyl l-phenylalanine trifluoroethyl esterC13H14NO3F3Ee >99.5%[α]D25=+44.7 (c 1.01, CHCl3)Source of chirality: N-acetyl l-phenylalanineAbsolute configuration (S)

N-Octanoyl l-alanineC11H21NO3Ee >99.5%[α]D25=+5.8 (c 1.00, CHCl3)Source of chirality: l-alanineAbsolute configuration (S)

N-Octanoyl l-alanine trifluoroethyl esterC13H22NO3F3Ee >99.5%[α]D25=-7.5 (c 1.03, CHCl3)Source of chirality: l-alanineAbsolute configuration (S)

(S)-2-Ethanamido-N-(4-phenylbutan-2-yl)ethanamideC14H20N2O2Ee >99.5%[α]D25=+3.9 (c 1.08, CHCl3)Source of chirality: enzymatic resolutionAbsolute configuration (S)

(S)-N-(2-oxo-2-(4-phenylbutan-2-ylamino)ethyl)benzamideC19H22N2O2Ee >99.5%[α]D25=+1.3 (c 0.83, CHCl3)Source of chirality: enzymatic resolutionAbsolute configuration (S)

(S,S)-N-(1-Oxo-3-phenyl-1-(4-phenylbutan-2-ylamino)propan-2-yl)octanamideC27H38N2O2Ee >99.5%[α]D25=-2.3 (c 0.83, CHCl3)Source of chirality: enzymatic resolutionAbsolute configuration (S,S)

(S,S)-2-Ethanamido-3-phenyl-N-(4-phenylbutan-2-yl)propanamideC21H26N2O2Ee >99.5%[α]D25=-3.4 (c 1.01, CHCl3)Source of chirality: enzymatic resolutionAbsolute configuration (S,S)

(S,S)-N-(1-Oxo-1-(4-phenylbutan-2-ylamino)propan-2-yl)octanamideC21H34N2O2Ee >99.5%[α]D25=-45.4 (c 1.00, CHCl3)Source of chirality: enzymatic resolutionAbsolute configuration (S,S)

(S)-N-(2-Oxo-2-(4-phenylbutan-2-ylamino)ethyl)-8-phenyloctanamideC26H36N2O2Ee >99.5%[α]D25=+3.1 (c 1.03, CHCl3)Source of chirality: enzymatic resolutionAbsolute configuration (S)

(S)-2-(Phenylamino)-N-(4-phenylbutan-2-yl)ethanamideC18H22N2OEe >99.5%[α]D25=+3.1 (c 1.21, CHCl3)Source of chirality: enzymatic resolutionAbsolute configuration (S)

(S)-N-(2-Oxo-2-(4-phenylbutan-2-ylamino)ethyl)-9-phenylnonanamideC27H38N2O2Ee >99.5%[α]D25=+2.1 (c 0.99 in CHCl3)Source of chirality: enzymatic resolutionAbsolute configuration (S)