title: Evaluation of acid-labile S-protecting groups to prevent Cys racemization in Fmoc solid-phase peptide synthesis.
abstract: Phosphonium and uronium salt-based reagents enable efficient and effective coupling reactions and are indispensable in peptide chemistry, especially in machine-assisted SPPS. However, after the activating and coupling steps with these reagents in the presence of tertiary amines, Fmoc derivatives of Cys are known to be considerably racemized during their incorporation. To avoid this side reaction, a coupling method mediated by phosphonium/uronium reagents with a weaker base, such as 2,4,6-trimethylpyridine, than the ordinarily used DIEA or that by carbodiimide has been recommended. However, these methods are appreciably inferior to the standard protocol applied for SPPS, that is, a 1 min preactivation procedure of coupling with phosphonium or uronium reagents/DIEA in DMF, in terms of coupling efficiency, and also the former method cannot reduce racemization of Cys(Trt) to an acceptable level (<1.0%) even when the preactivation procedure is omitted. Here, the 4,4'-dimethoxydiphenylmethyl and 4-methoxybenzyloxymethyl groups were demonstrated to be acid-labile S-protecting groups that can suppress racemization of Cys to an acceptable level (<1.0%) when the respective Fmoc derivatives are incorporated via the standard SPPS protocol of phosphonium or uronium reagents with the aid of DIEA in DMF. Furthermore, these protecting groups significantly reduced the rate of racemization compared to the Trt group even in the case of microwave-assisted SPPS performed at a high temperature.
authors.affiliation: SAITO Research Center, Peptide Institute, Inc., Ibaraki, Osaka, 567-0085, Japan.
authors.forename: Hajime
authors.initials: H
authors.lastname: Hibino
authors.forename: Yasuyoshi
authors.initials: Y
authors.lastname: Miki
authors.forename: Yuji
authors.initials: Y
authors.lastname: Nishiuchi
mesh_terms: D000595:Amino Acid Sequence; D000596:Amino Acids / Q000737:chemistry; D004126:Dimethylformamide / Q000737:chemistry; D005449:Fluorenes / Q000737:chemistry; D008872:Microwaves; D009943:Organophosphorus Compounds / Q000737:chemistry; D010455:Peptides / Q000138:chemical synthesis*; D010880:Piperidines / Q000737:chemistry; D060327:Solid-Phase Synthesis Techniques; D012997:Solvents / Q000737:chemistry; D013237:Stereoisomerism; D014230:Triazoles / Q000737:chemistry
chemical_list: D000596:Amino Acids; D005449:Fluorenes; C016456:N(alpha)-fluorenylmethyloxycarbonylamino acids; D009943:Organophosphorus Compounds; D010455:Peptides; D010880:Piperidines; D012997:Solvents; D014230:Triazoles; C496388:benzotriazol-1-yloxtri (pyrrolidinol)phosphonium hexafluorophosphate; C032727:piperidine; D004126:Dimethylformamide
keywords: 4,4′-dimethoxydiphenylmethyl (Ddm); 4-methoxybenzyloxymethyl (MBom); cysteine; microwave (MW)-assisted solid phase peptide synthesis (SPPS); phosphonium reagent; protecting group; racemization; uronium reagent