The adaptive potential of a survival artist: characterization of the in vitro interactions of Toxoplasma gondii tachyzoites with di-cationic compounds in human fibroblast cell cultures.
The impact of di-cationic pentamidine-analogues against Toxoplama gondii (Rh- and Me49-background) was investigated. The 72 h-growth assays showed that the arylimidamide DB750 inhibited the proliferation of tachyzoites of T. gondii Rh and T. gondii Me49 with an IC(50) of 0·11 and 0·13 μM, respectively. Pre-incubation of fibroblast monolayers with 1 μM DB750 for 12 h and subsequent culture in the absence of the drug also resulted in a pronounced inhibiton of parasite proliferation. However, upon 5-6 days of drug exposure, T. gondii tachyzoites adapted to the compound and resumed proliferation up to a concentration of 1·2 μM. Out of a set of 32 di-cationic compounds screened for in vitro activity against T. gondii, the arylimidamide DB745, exhibiting an IC(50) of 0·03 μM and favourable selective toxicity was chosen for further studies. DB745 also inhibited the proliferation of DB750-adapted T. gondii (IC(50)=0·07 μM). In contrast to DB750, DB745 also had a profound negative impact on extracellular non-adapted T. gondii tachyzoites, but not on DB750-adapted T. gondii. Adaptation of T. gondii to DB745 (up to a concentration of 0·46 μM) was much more difficult to achieve and feasible only over a period of 110 days. In cultures infected with DB750-adapted T. gondii seemingly intact parasites could occasionally be detected by TEM. This illustrates the astonishing capacity of T. gondii tachyzoites to adapt to environmental changes, at least under in vitro conditions, and suggests that DB745 could be an interesting drug candidate for further assessments in appropriate in vivo models.
Institute of Parasitology, Vetsuisse Faculty, University of Berne, Switzerland.
Christian
C
Kropf
Karim
K
Debache
Christoph
C
Rampa
Fabienne
F
Barna
Michelle
M
Schorer
Chad E
CE
Stephens
Mohamed A
MA
Ismail
David W
DW
Boykin
Andrew
A
Hemphill
D000578:Amidines / Q000494:pharmacology; D000818:Animals; D000981:Antiprotozoal Agents / Q000494:pharmacology; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D005347:Fibroblasts / Q000469:parasitology*; D005663:Furans / Q000494:pharmacology; D006801:Humans; D015394:Molecular Structure; D011725:Pyridines / Q000494:pharmacology; D014122:Toxoplasma / Q000502:physiology*; D014709:Vero Cells
C552045:2,5-bis(2-ethoxy-4-(2-pyridylimino)aminophenyl)furan; D000578:Amidines; D000981:Antiprotozoal Agents; C561940:DB 750; D005663:Furans; D011725:Pyridines