Thermitase – eine thermostabile Serinprotease. III. Synthese von N‐Acyl‐peptidmethylketonen als reversible Enzyminhibitoren

Abstract

Thermitase – A Thermostable Serine Protease. III. Synthesis of N‐Acylated Peptide Methyl Ketones as Inhibitors Reversibly Bound to the EnzymeMethyl ketone derivatives of dipeptides to pentapeptides are used as suitable tools in the investigation of the non‐covalent binding for subsite mapping of the active site of the enzyme. The synthesis is mainly performed by fragment condensation of N‐acylated peptides or amino acids with methyl ketone derivatives of amino acids. These are prepared from the Z‐protectedAbkürzungen: Ac = Acetyl, Z = Benzyloxycarbonyl, Boc = t‐Butyloxycarbonyl, Suc = Succinyl, Glp = Pyroglutamyl, OBz = Benzylester, pNA = p‐Nitranilid, CH3 = Methylketon, CH2Cl = Chlormethylketon, MA = Mischanhydridmethode, TEA = Triethylamin, NEM = N‐Ethylmorpholin, CKEE = Chlorkohlensäureethylester, CKBE = Chlorkohlensäurebutylester, MeOH = Methanol, EtOH = Ethanol, AcOH = Essigsäure, Ac2O = Acetanhydrid, Etac = Ethylacetat, THF = Tetrahydrofuran, E = Ether, PE = Petrolether, DMF = Dimethylformamid, DMSO = Dimethylsulfoxid, DMAP = 4‐(Dimethylamino)‐pyridin cloromethyl ketones by catalytic hydrogenation. For the coupling steps the Z‐protection is preferred. The peptide methyl ketones used in kinetic studies were N‐protected by the Z, acetyl, Boc or pyroglutamyl residue.