Identification and characterization of an endo/exonuclease in Pneumocystis carinii that is inhibited by dicationic diarylfurans with efficacy against Pneumocystis pneumonia.
Dicationic diarylfurans and dicationic carbazoles are under development as therapeutic agents against opportunistic infections. While their ability to bind to the minor groove of DNA has been established, the complete mechanism of action has not. We demonstrate here that an effective diarylfuran, 2,5-bis[4-(N-isopropylguanyl)phenyl]furan, inhibits an endo/exonuclease activity present in Pneumocystis carinii, Cryptococcus neoformans, Candida albicans, and Saccharomyces cerevisiae. This activity was purified from the particulate fraction of P. carinii. The enzyme requires Mg++ or Mn++, and shows preferences for single-over double stranded DNA and for AT-rich over GC-rich domains. A panel of 12 dicationic diarylfurans and eight dicationic carbazoles, previously synthesized, were evaluated for inhibition of the purified nuclease and for efficacy against Pneumocystis pneumonia in rats. Among the diarylfurans, potency of nuclease inhibition, in vivo antimicrobial activity, and DNA binding strength were all strongly correlated (p < 0.001). These findings suggest that one target for antimicrobial action of the diarylfurans may be a nucleolytic or other event requiring unpairing of DNA strands. Dicationic carbazoles which were strong nuclease inhibitors all displayed anti-Pneumocystis activity in vivo, but there were also noninhibitory carbazoles with in vivo efficacy.
Hildebrandt
E
E
Department of Pathobiology, College of Veterinary Medicine, Auburn University, Alabama 36849, USA.
Boykin
D W
DW
Kumar
A
A
Tidwell
R R
RR
Dykstra
C C
CC
D000818:Animals; D000935:Antifungal Agents / Q000627:therapeutic use*; D001550:Benzamidines / Q000627:therapeutic use; D002227:Carbazoles / Q000627:therapeutic use; D002413:Cations, Divalent / Q000627:therapeutic use; D003851:Deoxyribonucleases / Q000037:antagonists & inhibitors* / Q000302:isolation & purification; D004706:Endodeoxyribonucleases / Q000037:antagonists & inhibitors; D005090:Exodeoxyribonucleases / Q000037:antagonists & inhibitors; D005663:Furans / Q000627:therapeutic use; D006358:Hot Temperature; D011010:Pneumocystis / Q000201:enzymology*; D011020:Pneumonia, Pneumocystis / Q000188:drug therapy*; D011489:Protein Denaturation; D051381:Rats; D013379:Substrate Specificity
D000935:Antifungal Agents; D001550:Benzamidines; D002227:Carbazoles; D002413:Cations, Divalent; D005663:Furans; D003851:Deoxyribonucleases; D004706:Endodeoxyribonucleases; D005090:Exodeoxyribonucleases; C032157:benzamidine