Complete and unambiguous assignments of the 1H- and 13C-nmr spectra of the potent antitumor antibiotic staurosporine have been accomplished using a combination of one- and two-dimensional nmr techniques, including one-bond and long-range heteronuclear multiple-quantum coherence spectroscopy (HMQC). Staurosporine was found to be highly cytotoxic in the KB and P-388 assays but was inactive in the microtubulin assembly assay.