Evaluation of the cytotoxic mechanisms mediated by the broad-spectrum antitumor alkaloid acronycine and selected semisynthetic derivatives.
Acronycine (I) is a broad-spectrum antitumor agent whose development as a clinically useful agent has been hindered, in part, due to its poor solubility characteristics. With the goal of acquiring information that may prove of value in the development of structurally related compounds of greater clinical utility, mechanistic studies were performed with acronycine (I) and two semisynthetic derivatives, 2-nitroacronycine (II) and acronycine azine (III). These three substances demonstrated cytotoxic activity with several human tumor cell lines (breast, colon, lung, melanoma, KB-3, and drug-resistant KB-V1). Compounds II and III demonstrated greater activity than I, and more detailed studies were performed with cultured human breast cancer cells (UISO-BCA-1). Acronycine azine (III) induced the cells to accumulate in the G0/G1 phase of the cell cycle. It effectively inhibited the in vitro catalytic activities of partially purified DNA and RNA polymerases in a manner that was competitive with respect to DNA substrate. As judged by spectrophotometric titration, compound III interacted with calf thymus DNA, calf liver RNA, and a variety of single- and double-stranded (deoxy)ribonucleotides. Although no nucleic acid base specificity was discernable, this interaction appeared to be related to the cytotoxic mechanism of this dimeric substance. Monomeric compounds I and II did not interact with nucleic acids, but were effective inhibitors of DNA and RNA synthesis as judged by in vitro systems comprised of cultured UISO-BCA-1 cells or homogenates derived from these cells. The relative inhibitory activities of compounds I and II correlated with their cytotoxic activities suggesting a causal relationship. In addition, these two compounds induced cultured cells to accumulate in the phase of the cell cycle wherein the DNA content ranged from 2n-4n (S + G2/M), and inhibited in vitro DNA and RNA synthesis in a manner that was competitive with respect to nucleotide (TTP or UTP) substrate. Compounds I and II demonstrated greater cytotoxic activity with drug-resistant KB-V1 cells as compared with the parent (drug-sensitive) cell line, whereas this was not the case with compound III. Based on these results and previous literature reports, compounds I, II and III are likely to function by multiple mechanisms of action. However, it appears that alteration of nucleic acid metabolism is key to the activity of each of the substances.
Shieh
H L
HL
Program for Collaborative Research in the Pharmaceutical Sciences, College of Pharmacy, University of Illinois, Chicago.
Pezzuto
J M
JM
Cordell
G A
GA
D000175:Acronine / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000494:pharmacology*; D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology*; D001943:Breast Neoplasms / Q000473:pathology; D002453:Cell Cycle / Q000187:drug effects; D002460:Cell Line; D002467:Cell Nucleus / Q000201:enzymology; D002470:Cell Survival / Q000187:drug effects; D004261:DNA Replication / Q000187:drug effects; D012321:DNA-Directed RNA Polymerases / Q000037:antagonists & inhibitors / Q000302:isolation & purification; D004354:Drug Screening Assays, Antitumor; D005260:Female; D006801:Humans; D007700:Kinetics; D007930:Leucine / Q000378:metabolism; D008175:Lung Neoplasms / Q000473:pathology; D008297:Male; D008545:Melanoma / Q000473:pathology; D051379:Mice; D008819:Mice, Nude; D009363:Neoplasm Proteins / Q000096:biosynthesis; D009368:Neoplasm Transplantation; D012334:RNA, Neoplasm / Q000096:biosynthesis; D013936:Thymidine / Q000378:metabolism; D014410:Tumor Stem Cell Assay; D014529:Uridine / Q000378:metabolism
D000970:Antineoplastic Agents; D009363:Neoplasm Proteins; D012334:RNA, Neoplasm; D012321:DNA-Directed RNA Polymerases; D007930:Leucine; D000175:Acronine; D013936:Thymidine; D014529:Uridine