Expedient solid-phase synthesis of fluorogenic protease substrates using the 7-amino-4-carbamoylmethylcoumarin (ACC) fluorophore.
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.
Maly
Dustin J
DJ
Department of Chemistry, University of California, Berkeley, California 94720, USA.
Leonetti
Francesco
F
Backes
Bradley J
BJ
Dauber
Deborah S
DS
Harris
Jennifer L
JL
Craik
Charles S
CS
Ellman
Jonathan A
JA
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
C411664:7-amino-4-carbamoylmethylcoumarin; D000596:Amino Acids; D003374:Coumarins; D005456:Fluorescent Dyes; D005341:Fibrinolysin; D016333:HIV Protease