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Cancer is one of the leading causes of death in the United States and Europe. Of the cancers, bladder cancer is the 10th most frequently diagnosed cancer and the 13th most frequently diagnosed cancer in men. There are many studies showing that proteolytic enzymes, e.g. A Disintegrin and Metalloproteinases (ADAMs), play a key role in the development and progression of neoplasms. In this paper, we present the use of chromogenic substrate of ADAM15 for the qualitative determination of specific activity of enzyme in urine of patients with confirmed bladder cancer. In the first step, we optimized the substrate molecule in non-primed positions using combinatorial chemistry. By means of the obtained ABZ-His-Ala-Arg-Gly-ANB-NH 2 peptide, we detected ADAM15 activity in urine samples collected from patients diagnosed with bladder cancer. In contrast, we did not observe such activity in urine obtained from healthy volunteers. [Display omitted] • Bladder cancer. • ADAM15. • Urinary enzymes. • Chromogenic substrates. [ABSTRACT FROM AUTHOR]