title: Expedient solid-phase synthesis of fluorogenic protease substrates using the 7-amino-4-carbamoylmethylcoumarin (ACC) fluorophore.
abstract: A highly efficient solid-phase synthesis method for the preparation of fluorogenic protease substrates based upon the bifunctional leaving group 7-amino-4-carbamoylmethylcoumarin (ACC) is reported. Methods for the large-scale preparation of the novel fluorogenic leaving-group ACC are provided (Scheme 1). Detailed procedures are also provided for loading a diverse set of amino acids to support-bound ACC in good yields and with minimal racemization. Finally, procedures are included for the preparative synthesis of optimized ACC substrates for HIV-1 protease and plasmin.
authors.affiliation: Department of Chemistry, University of California, Berkeley, California 94720, USA.
authors.forename: Dustin J
authors.initials: DJ
authors.lastname: Maly
authors.forename: Francesco
authors.initials: F
authors.lastname: Leonetti
authors.forename: Bradley J
authors.initials: BJ
authors.lastname: Backes
authors.forename: Deborah S
authors.initials: DS
authors.lastname: Dauber
authors.forename: Jennifer L
authors.initials: JL
authors.lastname: Harris
authors.forename: Charles S
authors.initials: CS
authors.lastname: Craik
authors.forename: Jonathan A
authors.initials: JA
authors.lastname: Ellman
mesh_terms: D000596:Amino Acids / Q000737:chemistry; D003374:Coumarins / Q000737:chemistry / Q000378:metabolism*; D005341:Fibrinolysin / Q000378:metabolism*; D005456:Fluorescent Dyes / Q000737:chemistry / Q000378:metabolism*; D016333:HIV Protease / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy; D013058:Mass Spectrometry; D013379:Substrate Specificity
chemical_list: C411664:7-amino-4-carbamoylmethylcoumarin; D000596:Amino Acids; D003374:Coumarins; D005456:Fluorescent Dyes; D005341:Fibrinolysin; D016333:HIV Protease