Molecular docking, structure-activity relationship and biological evaluation of the anticancer drug monastrol as a pteridine reductase inhibitor in a clinical isolate of Leishmania donovani.
OBJECTIVE
Using the pteridine reductase (PTR1) enzyme of Leishmania as the target, the objective of our study was to find a drug candidate that can enter the clinical development process after being evaluated for safety and efficacy in animals.


METHODS
Monastrol (R) and (S) enantiomers were docked using the QUANTUM program into the active site of a Leishmania donovani PTR1 (LdPTR1) homology model. A structure-activity relationship based on a homology model of a recombinant enzyme was substantiated by a recombinant enzyme inhibition assay. We adapted an L. donovani (transfected with green fluorescent protein) intramacrophage amastigote screening assay as a cellular model for leishmaniasis. Furthermore, since the clinicopathological features and immunopathological mechanisms of visceral leishmaniasis (VL) in a hamster model are remarkably similar to those of human disease, systemic infection of hamsters with L. donovani was utilized to collect in vivo data for monastrol.


RESULTS
Both monastrol (R) and (S) enantiomers fit well in the ligand-binding pocket of LdPTR1. Monastrol exhibits a K(i) value of 0.428 microM in the recombinant enzyme inhibition assay. We confirm monastrol as a potent inhibitor of PTR1 in Leishmania; it inhibits proliferation of amastigotes with an IC(50) (50% inhibitory concentration) of 10 microM in macrophage cultures infected with an L. donovani clinical isolate, with no host cytotoxicity. We also show that in experimental animals, oral administration of a 5 mg/kg dose of monastrol on two alternate days inhibits 50% of parasite growth, giving therapeutic backing to the use of monastrol as a potent antileishmanial in human VL cases.


CONCLUSIONS
To our knowledge, this is the first report presenting monastrol as a potent oral antileishmanial.
Kaur
Jaspreet
J
Drug Target Discovery & Development Division, Central Drug Research Institute, Chattar Manzil Palace, PO Box No. 173, Lucknow-226001, CSIR, India.
Sundar
Shyam
S
Singh
Neeloo
N
D000284:Administration, Oral; D000818:Animals; D000981:Antiprotozoal Agents / Q000008:administration & dosage / Q000494:pharmacology*; D006224:Cricetinae; D004195:Disease Models, Animal; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D020128:Inhibitory Concentration 50; D007893:Leishmania donovani / Q000187:drug effects* / Q000302:isolation & purification; D007898:Leishmaniasis, Visceral / Q000188:drug therapy / Q000469:parasitology; D008264:Macrophages / Q000469:parasitology; D008297:Male; D008647:Mesocricetus; D056004:Molecular Dynamics Simulation; D010088:Oxidoreductases / Q000037:antagonists & inhibitors* / Q000737:chemistry; D021261:Parasitic Sensitivity Tests; D011743:Pyrimidines / Q000737:chemistry / Q000494:pharmacology*; D013329:Structure-Activity Relationship; D013871:Thiones / Q000737:chemistry / Q000494:pharmacology*
D000981:Antiprotozoal Agents; D004791:Enzyme Inhibitors; D011743:Pyrimidines; D013871:Thiones; C400223:monastrol; D010088:Oxidoreductases; C085592:pteridine reductase