The HIV − 1 protease inhibitor Amprenavir targets Leishmania donovani topoisomerase I and induces oxidative stress-mediated programmed cell death

Roy, Amit; Behera, Sachidananda; Mazire, Priyanka H.; Kumari, Bhavini; Mandal, Arabinda; Purkait, Bidyut; Ghosh, Payel; Das, Prolay; Das, Pradeep

doi:10.1016/j.parint.2021.102287

Cited by 12 publications

(11 citation statements)

References 32 publications

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“…ROS are highly reactive and modify the proteins, lipids, and nucleic acids. Therefore, ROS-mediated cell damage is a frequent event in apoptosis. ,, Our results, indicating that holanamine causes oxidative stress by inducing ROS generation, are supported by prior research showing that a number of natural products trigger ROS formation inside parasites, which has detrimental effects on them …”

Section: Resultssupporting

confidence: 85%

“…Therefore, ROS-mediated cell damage is a frequent event in apoptosis. 23,25,26 Our results, indicating that holanamine causes oxidative stress by inducing ROS generation, are supported by prior research showing that a number of natural products trigger ROS formation inside parasites, which has detrimental effects on them. 27 Holanamine Triggers the Externalization of Phosphatidylserine, Which Causes Apoptosis-like Cell Death in Leishmania Cells.…”

Section: ■ Results and Discussionsupporting

confidence: 83%

“…The control cells exhibited weak fluorescence, whereas the holanamine (IC 50 dose)-treated cells exhibited 75, 87, and 96% of parasites having positive for superoxide radicals at 8, 16, and 24 h, respectively, indicative of severe oxidative stress (Figure b,c). In the previous reports, it has been reported that increased cellular ROS production is responsible for the depolarization of mitochondrial membrane potential (ΔΨ m ) and subsequent cell death. ,, This study suggests that the loss of ΔΨ m could be due to oxidative stress inducing parasitic death via an apoptosis-like mechanism.…”

Section: Resultsmentioning

confidence: 72%

“…Changes in the mitochondrial inner membrane electrochemical potential (ΔΨ m ) in living cells are detected by using the TMRM (tetramethylrhodamine, methyl ester) dye. Uptake of TMRM by mitochondria may be utilized as an effective distinction between apoptotic and healthy cells . After treatment with holanamine, it was observed that the mitochondrial membrane potential was significantly changed, indicating the depolarization of mitochondrial membrane potential.…”

Section: Resultsmentioning

confidence: 99%

“…Molecular docking analysis was performed using AUTODOCK Vina software . The crystal structure of LdTop1B in complex with the ligand vanadate was obtained from the Protein Data Bank (PDB ID: 2B9S) and preprocessed appropriately for molecular docking study following the computational protocol reported by Saha et al and Roy et al , In this protocol, the core domain of topoisomerase structure has been used as a target for the docking simulations. A site known to have important residues (Arg-314, Lys-352, and Arg-410) can only be exposed by removing the DNA.…”

Section: Materials and Methodsmentioning

confidence: 99%

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Holanamine, a Steroidal Alkaloid from the Bark of Holarrhena pubescens Wall. ex G. Don Inhibits the Growth of Leishmania donovani by Targeting DNA Topoisomerase 1B

et al. 2022

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Leishmaniasis is a group of neglected tropical diseases (NTDs) caused by about 20 species of obligate intracellular protozoan parasites of the genus Leishmania, which occurs in cutaneous, mucocutaneous, and visceral forms. Many researchers have sought to utilize natural products for novel and effective treatments to combat many infectious diseases, including leishmaniasis. Holarrhena pubescens Wall. ex G. Don (Apocynaceae) bark is a rich source of bioactive steroidal alkaloids. The total alkaloidal extract (IC50 6.12 ± 0.117 μg/mL), and the isolated alkaloid, holanamine, showed significant antileishmanial activity (IC50 2.66 ± 0.112 μM against AG83 and 3.80 ± 0.126 μM against BHU-575) against the Leishmania donovani parasite, better than miltefosine (IC50 19.61 ± 0.093 μM against AG83 and 23.20 ± 0.094 μM against BHU-575). Holanamine inhibited the L. donovani topoisomerase 1B (LdToP1B) in a non-competitive manner (IC50 2.81 ± 0.105 μM), indicating that it interacts with the free enzyme and enzyme–DNA complex without inhibiting human topoisomerase. Hydrogen bonding and hydrophobic interactions of holanamine with the N-terminal and hinge region of the large subunit of LTop1B is responsible for its potent antileishmanial activity, as shown by docking studies. Treatment with holanamine causes apoptotic-like cell death by generating cellular and mitochondrial reactive oxygen species, disrupting the mitochondrial membrane potential and inducing ultrastructural alterations in the promastigotes. Holanamine effectively clears intracellular amastigotes but minimally affects host macrophages with no significant cytotoxicity in HEK 293 and L929 cell lines. Thus, our studies show that holanamine can further be used to develop effective antileishmanial agents against evolving drug-resistant parasites.

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