SUCCESSFUL TREATMENT OF HUMAN CHRONIC LYMPHOCYTIC-LEUKEMIA XENOGRAFTSWITH COMBINATION BIOLOGICAL AGENTS AURISTATIN-PE AND BRYOSTATIN-1

We tested the activity of dolastatin 10 (a natural product derived fro m the shell-less marine mollusk, Dolabella auricularia, a sea hare) an d its structural modification, auristatin PE, alone and in combination with bryostatin 1 (a protein kinase C activator derived from the mari ne bryozoan Bugula neritina) on a human B-cell chronic lymphocytic leu kemia cell line (WSU-CLL) and in a severe combined immune deficient (S CID) mouse xenograft model bearing this cell line. WSU-CLL cells were cultured in RPMI 1640 at a concentration of 2 x 10(5)/ml using a 24-we ll plate. Agents were added to triplicate wells, and cell count, viabi lity, mitosis, and apoptosis were assessed after 24 h of incubation at 37 degrees C, Results showed that dolastatin 10 had no apparent inhib ition of cell growth at concentrations less than 500 pg/ml, Auristatin PE, on the other hand, showed significant growth inhibition at concen trations as low as 50 pg/ml, Auristatin PE-treated cultures, at this c oncentration, exhibited 27 and 4.5% mitosis and apoptosis, respectivel y. Dolastatin 10, at the same concentration, did not exert any effect and was comparable with that of control cultures. In the WSU-CLL-SCID mouse xenograft model, the efficacy of these agents alone and in combi nation with bryostatin 1 was evaluated. Tumor growth inhibition (T/C), tumor growth delay (T-C), and log(10) kill for dolastatin 10, aurista tin PE, and bryostatin 1 were 14%, 25 days, and 1.98; 2%, 25 days, and 1.98; 19%, 13 days, and 1.03, respectively. Auristatin-PE produced cu re in three of five mice, whereas dolastatin 10 showed activity but no cures. When given in combination, auristatin PE + bryostatin 1-treate d animals were all free of tumors (five of five) for 150 days and were considered cured. Dolastatin 10 + bryostatin 1-treated animals produc ed cure in only two of five mice. We conclude that: (a) auristatin-PE is more effective in this model than dolastatin 10; (b) auristatin PE can be administered at a concentration 10 times greater than dolastati n 10; (c) there is a synergetic effect between these agents and bryost atin 1, which is more apparent in the bryostatin 1 + auristatin PE com bination. The use of these agents should be explored clinically in the treatment of CLL.