Against this backdrop, a research team from the ICMR National Institute of Traditional Medicine in Belagavi, working with colleagues at KLE College of Pharmacy, has turned to a plant with deep roots in traditional medicine. Plumbago zeylanica, known in Ayurvedic practice as chitraka, has long been used across the Indian subcontinent for wound healing and inflammatory conditions, and folk accounts have credited it with value against snakebite. The new study, published in the journal 3 Biotech, set out to test whether those traditional claims could survive rigorous modern scrutiny, using a hydromethanolic extract of the plant’s aerial parts against cobra venom in a combined program of laboratory assays, animal experiments and computational modeling.

The in vitro results were striking in their breadth. The extract, abbreviated PZE by the researchers, inhibited phospholipase A2 activity in a dose-dependent fashion, with inhibition ranging from roughly 13 percent at lower concentrations to more than 53 percent at the highest dose tested. Because phospholipase A2 is a principal driver of venom-induced hemolysis, the team also examined its effect on red blood cell destruction and found that the extract reduced PLA2-mediated hemolysis by 46.2 percent. These are not trivial numbers for a crude plant extract, and they suggest that compounds within the aerial parts of the plant can interfere directly with one of the most destructive enzymatic families in elapid venom.

The enzyme-inhibitory profile extended well beyond phospholipase A2. PZE suppressed venom acetylcholinesterase activity by 58.12 percent, an effect with direct relevance to the neurotoxic component of cobra envenomation, since acetylcholinesterase breaks down the neurotransmitter that carries signals from nerve to muscle. Protease activity, which contributes to tissue degradation and hemorrhage, was inhibited by 81.94 percent, the strongest effect recorded in the enzymatic panel. A fluorometric assay also indicated apparent inhibition of collagenase, an enzyme that attacks the collagen scaffolding of connective tissue. Perhaps most tellingly, the extract preserved the alpha chain of fibrinogen, the blood protein that venoms frequently fragment to disrupt clotting, indicating protection of a key substrate at the heart of venom-induced coagulopathy.

Translating these findings into living systems, the researchers challenged mice with twice the lethal dose of cobra venom and treated them orally with the extract at 100 and 200 milligrams per kilogram of body weight. Untreated animals succumbed rapidly, but the treated groups survived significantly longer, with mean survival times of 16 hours 23 minutes and 19 hours 42 minutes at the low and high doses respectively. For comparison, the commercial antisnake venom serum, administered as the current standard of care, provided complete protection for the full 24-hour observation period. The plant extract did not outperform antivenom, and the authors are explicit on this point, but the demonstration that an orally delivered plant preparation can meaningfully extend survival against a supralethal venom challenge is a meaningful proof of concept for an adjunctive therapy that could be administered early, even before a patient reaches a clinic.

The in vivo work also probed the inflammatory and tissue-level consequences of envenomation. At a sublethal challenge of half the lethal dose, venom did not produce significant alterations in the measured serum biomarkers and biochemical parameters, but treatment with the extract reduced venom-associated elevations in interleukin-6 and tumor necrosis factor-alpha, two central cytokines of the inflammatory response, as well as the brain and cardiac isoforms of creatine kinase, enzymes whose release signals damage to neural and muscle tissue. Histopathological examination told a consistent story: venom produced dilatation of the central vein, vascular congestion and mild lymphocytic infiltration in the liver, along with vascular congestion and tubular hemorrhage in the kidney, and the extract markedly ameliorated all of these changes. The pattern suggests that PZE acts not only on venom enzymes themselves but also on the downstream inflammatory storm that venom triggers in host tissues.

To understand which molecular targets might underlie these effects, the team employed network pharmacology, a computational approach that maps the interactions between the phytochemicals in the extract and the host proteins involved in disease pathways. The analysis identified multiple host targets connected through inflammatory, immune and neuronal signaling networks, providing a systems-level rationale for why a crude extract containing many different compounds could produce such broad protective effects. This multitarget character is precisely what conventional single-molecule drug development struggles to achieve against a venom that is itself a complex cocktail of toxins, and it mirrors the logic by which polyvalent antivenoms work, albeit through entirely different molecular means.

The computational arm of the study went further, using molecular docking and molecular dynamics simulations to examine how individual constituents of the extract interact with venom enzymes at atomic resolution. Two flavonoid compounds, 6-C-fucosylluteolin and 6-C-galactosylluteolin, emerged as the most interesting candidates. Docking placed both molecules within the catalytic sites of acetylcholinesterase and phospholipase A2, and the molecular dynamics simulations demonstrated that these interactions remained stable over the simulated timescales, indicating that the binding is not a fleeting artifact but a physically plausible mode of enzyme inhibition. While docking and simulation cannot by themselves prove activity in a patient, they provide a mechanistic scaffold that connects the observed biochemical inhibition to specific compounds that can now be isolated, tested and potentially optimized.

The authors position PZE not as a replacement for antivenom but as a candidate adjunctive phytotherapeutic intervention for the early management of snakebite envenomation, to be used alongside conventional therapy. The logic is compelling. Antivenom must be administered in a clinical setting, is expensive, carries a risk of adverse reactions including anaphylaxis, and is often unavailable in the rural areas where bites occur. A plant-derived oral preparation that could blunt local tissue destruction and dampen inflammation during the critical window before antivenom is given would address a genuine gap in the treatment pathway, particularly in South Asia, where the socioeconomic burden of snakebite is enormous and where traditional plant remedies already occupy a trusted place in first-response practice.

Considerable work remains before any such therapy could reach patients. The active constituents need to be fully characterized and standardized, dose-response relationships and safety margins must be established in accordance with internationally recognized toxicology guidelines, and the protective effects demonstrated here in mice must be validated against the biogeographical variation that is known to exist in cobra venom across India, a variation that already challenges the efficacy of existing antivenoms. Clinical efficacy, drug interactions and the pharmacokinetics of the relevant flavonoids in envenomated patients all remain open questions. Nevertheless, the study represents a rigorous, multi-level validation of a traditional remedy, combining classical enzyme assays, survival studies, histopathology, cytokine profiling, network pharmacology and molecular simulation into a single coherent case. In doing so, it transforms an ethnobotanical claim from folklore into a testable hypothesis, and it adds Plumbago zeylanica to the short but growing list of medicinal plants whose anti-venom potential has survived contact with modern experimental science.

Subject of Research: Evaluation of Plumbago zeylanica aerial part extract as a multitarget adjunctive therapy against Naja naja snake venom

Article Title: Anti-envenomation potential of Plumbago zeylanica aerial part extract against Naja naja: experimental validation and mechanistic insights

Article References: Anti-envenomation potential of Plumbago zeylanica aerial part extract against Naja naja: experimental validation and mechanistic insights. (n.d.). https://doi.org/10.1007/s13205-026-05067-5

Image Credits: AI Generated

DOI: 10.1007/s13205-026-05067-5

Keywords: snakebite, Naja naja, Plumbago zeylanica, antivenom, phospholipase A2, acetylcholinesterase, network pharmacology, molecular docking, phytotherapy, envenomation, flavonoids, traditional medicine