Biology subjects

Edge, R. J.

Publications and source records attributed to Edge, R. J..

4 recordsLinked to original sources

bioRxiv2025

A drug combinational therapy consisting of toxin phospholipase A2 and metalloproteinase inhibitors provides preclinical protection against North American Crotalid snakebite envenoming

BackgroundAcross North America an estimated 3,800-6,500 snakebite envenomings occur annually, resulting in 7-15 deaths and an unknown number of disfigurements and disabilities. Most bites are caused by Crotalid snake species. The variable diversity and toxin complexity of crotalid venoms presents a considerable challenge to developing broadly effective small molecule therapeutics to better treat snakebite in this region. MethodsWe evaluated the ability of three small molecule, toxin inhibiting, repurposed drugs to inhibit the venom activities of six medically important crotalid snake species (Agkistrodon contortrix, Crotalus atrox, C. adamanteus, C. horridus, C. scutulatus and Sistrurus miliarius). These drugs target two pathologically relevant venom toxin families, the snake venom metalloproteinases (SVMPs; marimastat and DMPS) and phospholipases A2 (PLA2s; varespladib), and venom inhibition was measured using in vitro enzymatic and phenotypic plasma coagulation assays. Thereafter we evaluated the efficacy of individual drugs and dual drug combinations in in vivo preclinical models of snakebite envenoming, using both preincubation and rescue model formats. ResultsIn vitro bioassays demonstrated that the selected small molecules showed potent inhibition of the enzymatic activity of different toxin families to the nanomolar (varespladib vs PLA2 and marimastat vs SVMP) or micromolar (DMPS vs SVMP) level. Three of the venoms had anticoagulant activity, which varespladib restored to normal coagulation profiles, suggesting this activity is mostly driven by PLA2 toxins. Preclinical experiments revealed that pre-incubation of representative venoms with single drugs was insufficient to completely protect against lethality, except for varespladib against C. scutulatus. Superior efficacy was observed when drugs were used in a combination approach, with the combination of marimastat and varespladib providing greatest protection against lethality in both pre-incubation and rescue models. ConclusionsVenom variation among snake species makes the development of generic snakebite therapeutics challenging. In this study we showed that while SVMP and PLA2 inhibiting drugs show inhibitory potency against diverse North American snake venoms, drug combinations consisting of an SVMP inhibitor together with a PLA2 inhibitor are required to confer broad in vivo protection against lethality caused by envenoming. This study highlights the potential long-term value of drug combinations as next-generation therapeutics for snakebite envenoming.

molecular biology↗

bioRxiv2022

Two snakebite antivenoms have potential to reduce Eswatini's dependency upon a single, increasingly unavailable product: results of preclinical efficacy testing

BackgroundSnakebite is a major public health concern in Eswatini, where treatment relies upon one antivenom - SAIMR Polyvalent. Although effective in treating snakebite, SAIMR Polyvalent is difficult to source outside its manufacturing country (South Africa) and is dauntingly expensive. We compared the preclinical venom-neutralising efficacy of two alternative antivenoms and SAIMR Polyvalent against the lethal and tissue-destructive effects of venoms from five species of medically important snakes using in vivo murine assays. The test antivenoms were Panafrican manufactured by Instituto Clodomiro Picado and PANAF manufactured by Premium Serums & Vaccines. Principal FindingsIn vivo murine preclinical studies identified both test antivenoms were equally or more effective than SAIMR Polyvalent at neutralising lethal and tissue-destructive effects of Naja mossambica venom. Both test antivenoms were less effective than SAIMR Polyvalent at neutralising the lethal effects of Bitis arietans, Dendroaspis polylepis, Hemachatus haemachatus and Naja annulifera venoms, but similarly effective at neutralising tissue damage induced by B. arietans and H. haemachatus venoms. In vitro immunological assays identified that IgG titres and toxin-specificities of the test antivenoms were comparable to SAIMR Polyvalent. Plasma clotting disturbances by H. haemachatus and N. mossambica were effectively neutralised by the test antivenoms, whereas SAIMR Polyvalent failed to neutralise this bioactivity of N. mossambica venom. The SVMP activity of B. arietans venom was equally reduced by all three antivenoms. The PLA2 activities of H. haemachatus and N. mossambica were effectively neutralised by all three antivenoms. ConclusionsPanafrican outperformed PANAF, though both were less poly-specifically effective than SAIMR Polyvalent. The efficacy of these antivenoms against the lethal and tissue-destructive effects of N. mossambica venom, the most common biting species in Eswatini, identify that Panafrican and PANAF antivenoms offer effective alternatives to SAIMR for the treatment of snakebite in Eswatini, and potentially for neighbouring countries. Author SummaryTreatment of snakebite in Eswatini is reliant upon a single antivenom (SAIMR Polyvalent) manufactured in South Africa. This highly effective product is increasingly difficult to source and is expensive - alternative/additional antivenoms are urgently required to improve patient outcomes following snake envenoming. Using murine preclinical venom toxicity and antivenom efficacy assays, we identified two alternative antivenoms whose venom-neutralising characteristics, while less poly-specifically effective than SAIMR Polyvalent, were as effective against the lethal and tissue-destructive effects of the most common biting snake in Eswatini - Naja mossambica. This murine data, already shared with the Eswatini Ministry of Health, supports and justifies human testing of these two antivenoms in Eswatini because increasing the availability of effective and affordable treatments could resolve the current medical dependency of Eswatini snakebite patients upon a single, rarely available and expensive product.

pharmacology and toxicology↗

bioRxiv2022

Exploring the utility of recombinantly expressed snake venom serine protease toxins as immunogens for generating experimental snakebite antivenoms

Snakebite is a neglected tropical disease that causes high rates of global mortality and morbidity. Although snakebite can cause a variety of pathologies in victims, haemotoxic effects are particularly common and are typically characterised by haemorrhage and/or venom-induced consumption coagulopathy. Despite polyclonal antibody-based antivenoms being the mainstay life-saving therapy for snakebite, they are associated with limited cross-snake species efficacy, as there is often extensive toxin variation between snake venoms, including those used as immunogens for antivenom production. This restricts the therapeutic utility of any antivenom to certain geographical regions. In this study, we explored the feasibility of using recombinantly expressed toxins as immunogens to stimulate focused, pathology-specific, antibodies to broadly counteract specific toxins associated with snakebite envenoming. Three snake venom serine proteases (SVSP) toxins, sourced from geographically diverse and medically important viper snake venoms were successfully expressed in HEK293F mammalian cells and used for murine immunisation. Analyses of the resulting antibody responses revealed that ancrod and RVV-V stimulated the strongest immune responses, and that experimental antivenoms directed against these recombinant SVSP toxins, and a mixture of the three different immunogens, extensively recognised and exhibited immunological binding towards a variety of native snake venoms. While the experimental antivenoms showed some reduction in abnormal clotting parameters stimulated by the toxin immunogens and crude venom, specifically reducing the depletion of fibrinogen levels and prolongation of prothrombin times, fibrinogen degradation experiments revealed they broadly protected against venom- and toxin-induced fibrinogenolytic functional activities. Overall, our findings further strengthen the case for the use of recombinant venom toxins as supplemental immunogens to stimulate focused and desirable antibody responses capable of neutralising venom-induced pathological effects, and therefore potentially circumventing some of the limitations associated with current snakebite therapies.

immunology↗

bioRxiv2022

In vitro and in vivo venom-inhibition assays identify metalloproteinase-inhibiting drugs as potential treatments for snakebite envenoming by Dispholidus typus

Snakebite envenoming affects more than 250,000 people annually in sub-Saharan Africa. Envenoming by Dispholidus typus (boomslang) results in venom induced consumption coagulopathy, whereby highly abundant prothrombin-activating snake venom metalloproteinases (SVMPs) consume clotting factors and deplete fibrinogen. The only available treatment for D. typus envenoming is the monovalent SAIMR Boomslang antivenom. Treatment options are urgently required because this antivenom is often difficult to source and, at $6,000/vial, typically unaffordable for most snakebite patients. We therefore investigated the in vitro and in vivo preclinical efficacy of four SVMP inhibitors to neutralise the effects of D. typus venom; the matrix metalloproteinase inhibitors marimastat and prinomastat, and the metal chelators dimercaprol and DMPS. The venom of D. typus exhibited an SVMP-driven procoagulant phenotype in vitro. Marimastat and prinomastat demonstrated equipotent inhibition of the SVMP-mediated procoagulant activity of the venom in vitro, whereas dimercaprol and DMPS showed considerably lower potency. However, when tested in preclinical murine models of envenomation, DMPS and marimastat demonstrated partial protection against venom lethality, demonstrated by prolonged survival times of experimental animals, whereas dimercaprol and prinomastat failed to confer any protection at the doses tested. The results presented here demonstrate that DMPS and marimastat show potential as novel small molecule-based therapeutics for D. typus snakebite envenomation. These two drugs have been previously shown to be effective against Echis ocellatus venom induced consumption coagulopathy (VICC) in preclinical models, and thus we conclude that marimastat and DMPS may be valuable early intervention therapeutics to broadly treat VICC following snakebite envenoming in sub-Saharan Africa.

pharmacology and toxicology↗