Patent ID: 6037137
Filing Date: 2000-03-14
Classification: C07K,C12Q,G01N

Abstract:
A fluorogenic composition for the detection of the activity of a protease, said composition having the formula: ##STR19## wherein, P is a peptide comprising a protease binding site for said protease, said binding site consisting of about 2 to about 8 amino acids;F.sup.1 and F.sup.2 are fluorophores and F.sup.1 is attached to the amino terminal amino acid and F2 is attached to the carboxyl terminal amino acid;S.sup.1 and S.sup.2, when present, are peptide spacers ranging in length from 1 to about 50 amino acids and S.sup.1, when present, is attached to the amino terminal amino acid and S.sup.2, when present, is attached to the carboxyl terminal amino acid;i, j, k, 1, m, n, o, p, q, and r are independently 0 or 1;aa.sup.1 and aa.sup.10 are independently selected from the group consisting of lysine, ornithine and cysteine;aa.sup.2, aa.sup.3, aa.sup.8, and aa.sup.9 are independently selected from the group consisting of Asp, Glu, Lys, Ornithine, Arg, Citulline, homocitrulline, Ser, homoserine, Thr, and Tyr;aa.sup.5, aa.sup.4, aa.sup.6, and aa.sup.7 are independently selected from the group consisting of proline, 3,4-dehydroproline, hydroxyproline, alpha aminoisobutyric acid and N-methyl alanine;X is selected from the group consisting of Gly, .beta.Ala, .gamma.Abu ,Gly-Gly, Ahx, .beta.Ala-Gly, .beta.Ala-.beta.Ala, .gamma.Abu-Gly, .beta.Ala-.gamma.Abu, Gly-Gly-Gly (SEQ ID NO:37), .gamma.Abu-.gamma.Abu, Ahx-Gly, .beta.Ala-Gly-Gly, Ahx-.beta.Ala, .beta.Ala-.beta.Ala-Gly, Gly-Gly-Gly-Gly, Ahx-.gamma.Abu, .beta.Ala-.beta.Ala-.beta.Ala, .gamma.Abu-.beta.Ala-Gly, .gamma.Abu-.gamma.Abu-Gly, Ahx-Ahx, .gamma.Abu-.gamma.Abu-.beta.Ala, and Ahx-Ahx-Gly;Y is selected from the group consisting of Gly, .beta.Ala, .gamma.Abu, Gly-Gly, Ahx, Gly-.beta.Ala, .beta.Ala-.beta.Ala, Gly-.gamma.Abu, .gamma.Abu-.beta.Ala, Gly-Gly-Gly, .gamma.Abu-.gamma.Abu, Gly-Ahx, Gly-Gly-.beta.Ala,.beta.Ala-Ahx, Gly-.beta.Ala-.beta.Ala, Gly-Gly-Gly-Gly (SEQ ID NO:37), .gamma.Abu-Ahx, .beta.Ala-.beta.Ala-.beta.Ala, Gly-.beta.Ala-.gamma.Abu, Gly-.gamma.Abu-.gamma.Abu, Ahx-Ahx, .beta.Ala-.gamma.Abu-.gamma.Abu, and Gly-Ahx-Ahx;when i is 1, S.sup.1 is joined to aa.sup.1 by a peptide bond through a terminal alpha amino group of aa.sup.1 ; andwhen r is 1, S.sup.2 is joined to aa.sup.10 by a peptide bond through a terminal alpha carboxyl group of aa.sup.10.