Gold(III) macrocycles: nucleotide-specific unconventional catalytic inhibitors of human topoisomerase I.
Topoisomerase IB (Top1) is a key eukaryotic nuclear enzyme that regulates the topology of DNA during replication and gene transcription. Anticancer drugs that block Top1 are either well-characterized interfacial poisons or lesser-known catalytic inhibitor compounds. Here we describe a new class of cytotoxic redox-stable cationic Au(3+) macrocycles which, through hierarchical cluster analysis of cytotoxicity data for the lead compound, 3, were identified as either poisons or inhibitors of Top1. Two pivotal enzyme inhibition assays prove that the compounds are true catalytic inhibitors of Top1. Inhibition of human topoisomerase IIα (Top2α) by 3 was 2 orders of magnitude weaker than its inhibition of Top1, confirming that 3 is a type I-specific catalytic inhibitor. Importantly, Au(3+) is essential for both DNA intercalation and enzyme inhibition. Macromolecular simulations show that 3 intercalates directly at the 5'-TA-3' dinucleotide sequence targeted by Top1 via crucial electrostatic interactions, which include π-π stacking and an Au···O contact involving a thymine carbonyl group, resolving the ambiguity of conventional (drug binds protein) vs unconventional (drug binds substrate) catalytic inhibition of the enzyme. Surface plasmon resonance studies confirm the molecular mechanism of action elucidated by the simulations.
School of Chemistry and Physics, University of KwaZulu-Natal , Private Bag X01, Scottsville, Pietermaritzburg, 3209, South Africa.
Kate J
KJ
Akerman
Alexander M
AM
Fagenson
Vidusha
V
Cyril
Michael
M
Taylor
Mark T
MT
Muller
Matthew P
MP
Akerman
Orde Q
OQ
Munro
D002384:Catalysis; D018360:Crystallography, X-Ray; D006046:Gold / Q000737:chemistry*; D006801:Humans; D047028:Macrocyclic Compounds / Q000737:chemistry*; D059004:Topoisomerase I Inhibitors / Q000737:chemistry*
D047028:Macrocyclic Compounds; D059004:Topoisomerase I Inhibitors; D006046:Gold