Venom is obtained from the various creatures in different ways. Snakes and funnel web spiders are milked for their venom. Stonefish, redback spider and box jellyfish antivenoms are made from venom extracted from the animal by dissection.

This may be a dangerous process. Small doses of venom or venom components are injected into the animal, and the dose gradually increased as the animal builds up a tolerance to the venom. In response to the introduction of the venom a foreign substance , the animal produces antibodies to the venom.

When the doses being injected are large, the amount of antibody produced is large. These antibodies are harvested by taking blood from the animals and separating out the antibodies, which are then fragmented and purified by a series of digestion and processing steps.

When injected into a patient, the binding sites on the antibody fragments bind to the venoms or venom components in the circulation and neutralize the activity of the venoms in the patient. Antivenoms have been made since the s. Australia was one of the first countries in the world to experiment with snake antivenoms, in , when Frank Tidswell commenced immunization of a former ambulance horse with tiger snake N.

scutatus venom. CSL Ltd is the sole manufacturer of antivenoms for human use in Australia. Australian antivenoms are amongst the best in the world, in terms of purity and adverse reaction rate. Identification of the offending snake will aid in the choice of the appropriate antivenom and alert clinicians to particular features characteristic of envenomation by that type of snake.

Identification of snakes by the general public or by hospital staff is frequently unreliable. Sometimes, the snake is not seen, or is only glimpsed in retreat. In these cases, a snakebite venom detection kit may be used.

CSL Snake Venom Detection Kit including contents and packaging. The many isoforms of ion channels further complicate their selective targeting If a toxin binds an unknown target, identification of the target is challenging owing to the technical difficulty of screening target pools.

The binding of PLA2s to such a membrane protein narrows its biodistribution, focusing its hydrolytic action on muscle tissues , However, even after many years of study, the identity of the myotoxic PLA2 pharmacologic target is still unknown.

When target identification is successful, determining the target structure and the target—toxin complex is still challenging. Knowing the 3D structure of the complex is a requisite to the understanding of molecular recognition, without which the design of toxinomimetics within a structure-based paradigm is not possible.

In this case, toxinomimicry has to resort to a ligand-based paradigm, supported by measurements of ligand affinity for toxin mutants, which is less efficient than structure-based drug design because it is rooted in less molecular information.

The path forward should entail, at least in part, a much deeper involvement of computational chemistry. The increase in computational power allows for the more exact implementation of physical principles, which, together with the greater involvement of deep learning and artificial intelligence, is powering advances in computational fields important for snake-venom-based chemistry and drug discovery, such as protein homology modelling , , , , , ; protein—protein docking , , , , , , , ; computational mutagenesis, in particular alanine scanning , , , , , ; and the determination of enzymatic mechanisms 88 , 89 , 90 , Computational chemistry can thus have a decisive role in speeding up the process of drug discovery based on snake venom toxins.

Computational chemistry can intervene whenever a toxin with the bioactivity of interest acts on an unknown target. Today, it is possible to assemble a database of biological targets for which the molecular structures have been determined by X-ray or cryogenic electron microscopy and homology modelling and then to screen the database according to toxin—target affinity.

In several cases, the uncertainties associated with homology modelling and docking do not allow for the identification of a single and robust target. Nevertheless, computational chemistry reduces the target pool to a set small enough to be feasible for experimental testing. It is also challenging to determine target—toxin complex geometries with atomic-level accuracy through computation alone , particularly when modelled structures are involved.

Despite this, computational chemistry can narrow down the target and toxin regions that contact each other to the point at which experimental mutagenesis and other techniques can be applied to provide the final atomic-level information. As an example, computational and experimental methodologies were used together to clarify the mechanism by which mambalgins inhibit ASICs , In summary, high-level computational chemistry has the power to advance target identification and target—toxin structural determination if conducted in synergy with experiments; together they could facilitate either the use of unmodified toxins or the modelling of toxin-based small ligands.

For the latter, traditional medicinal chemistry can be employed to reduce the toxins into small, synthetic, bioavailable molecules while keeping most of the determinants for recognition and affinity. In modern Western civilization, the snake represents deceit and triggers both fascination and fear.

However, ancient civilizations respected the snake owing to the healing power of its venom. It is becoming evident that the ancients were right, as the venom of this splendid animal is an extraordinary library of bioactive compounds that has great medicinal potential.

Efforts to elucidate the chemical reactivity of the principal toxins within venom is helping to increase understanding of how toxins act on their prey targets, and how one can engineer toxin action to achieve a therapeutic goal.

Furthermore, understanding of venom chemistry allows for the rational design of transition-state small-molecule analogue inhibitors for primary enzymatic toxins that are today the most promising candidates for replacing the difficult-to-manage and expensive antibody-based treatments for snakebite envenoming.

The molecular recognition features of snake venom toxins are also being explored at a molecular level. The drugs already approved and under development derived from snake venom demonstrate that toxic bioactivity can be transformed into a therapy for the right disease.

Large toxin molecules can be redesigned and reduced to their recognition motifs for oral delivery while maintaining affinity and specificity. Of the many drugs in preclinical development, mambalgins in particular reflect the contrast between their therapeutic promise in this case, to relieve pain and their origin from one of the most feared snakes on the planet.

In terms of the future of venom-based drug development, we assert that toxinomimicry is an exciting alternative and a complement to the use of unmodified toxins. Furthermore, computational chemistry, which is still underused in the field, can accelerate the understanding of snake venom chemistry and hence the development of new drugs.

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There is a growing interest in the development of recombinant antivenoms 55 , This involves, for example, the preparation of animal- or human-derived monoclonal antibodies against the lethal components of venoms. Proofs of concept of this strategy have been published 57 , One major requirement of this approach is that the major lethal toxin s of the venom must be identified and used for antibody selection.

When more than one toxin is relevant in a particular venom, there is a need to generate additional antibodies for a successful neutralization. Since these antibodies are produced against one or few toxins, a challenging issue for this strategy is to ensure the neutralization of heterologous toxins present in other venoms.

It should be possible to further increase the para-specificity of the antiserum by including additional venom toxin fractions in the immunization mix.

For example, inclusion of toxin fractions of some African mamba venoms D. angusticeps and D. viridis and some American coral snakes e. could increase the neutralizing scope of the antiserum.

By carefully selecting the venoms and fractions to be added to the immunizing mix it should be possible to expand the scope of coverage of neurotoxic venoms, ideally to neutralize the most important elapid venoms in the world. Snakebite envenomation is a WHO classified category A Neglected Tropical Diseases, i.

a disease of highest importance. One of the four pillars of this strategy is to ensure safe and effective treatments, particularly referring to antivenoms, which represent the only scientifically-validated therapy for these envenomings.

As shown in this work, a pan-specific antivenom against neurotoxic venoms would be a powerful therapeutic tool to save lives of people suffering these envenomings in different parts of the world, by neutralizing a wide spectrum of neurotoxic snake venoms which otherwise require region- or species-specific antivenoms for treatment.

The antiserum exhibited a wide para-specificity by neutralizing at least 36 neurotoxic venoms of snakes of 10 genera from four continents. The pool of diverse toxin antigens in the immunogen mix enabled the production of diverse antibody paratopes, which facilitate the interaction of the antibodies with the epitopes of various neurotoxins from homologous as well as heterologous snake venoms.

Dendroaspis polylepis, D. angusticeps, D. viridis and Naja senegalensis venoms were obtained from Latoxan Valence, France ; Laticauda colubrina, Pseudechis australis, Oxyuranus scutellatus and Notechis scutatus venoms were obtained from Venom Supplies Pty Ltd Australia.

Hydrophis schistosus venom was provided by Dr. CH Tan, and Micrurus nigrocinctus venom was provided by Prof. José María Gutiérrez.

These two venoms were obtained from several specimens kept in captivity M. nigrocinctus or captured wild H. Naja kaouthia Thailand principal post-synaptic neurotoxin 3 NK3 was purified as described by Karlsson et al. The pan-specific antiserum used in the present study was from the same batch as that obtained from horses immunized with mixtures of venoms and venom fractions 11 using the protocol briefly described below.

All chemicals and biochemical were from Sigma Chemical Co. St Louis, Missouri, USA unless otherwise stated. Experiments carried out in horses regarding care, bleeding and immunization were approved by the Animal Care and Use Committee of the Faculty of Veterinary Science, Mahidol University, Protocol and clearance no.

MUVS in accordance with the Guidelines of the National Research Council of Thailand. Preparation of the pan-specific antiserum was described previously and Bungarus spp. inhabiting different geographical locations of Asia The toxin fractions of Naja spp.

The presence of α-neurotoxins in venoms was estimated by the venom-mediated inhibition of the binding of purified nAChR to immobilized elapid post-synaptic neurotoxins, as described previously californica electroplax.

The amount of nAChR bound to the immobilized NK3 in the wells was estimated by adding rat anti-nAChR serum at dilution followed by a dilution of goat anti-rat IgG-enzyme conjugated HRP and enzyme substrate.

If the tested neurotoxic venom contained α-neurotoxin which could specifically interact with nAChR, the percent binding of the receptor to the NK3 immobilized plate is reduced and can be calculated using the following formula:. Venom lethality median lethal dose, LD 50 and the median effective doses ED 50 of the pan-specific antiserum against the venoms tested were determined and analyzed as previously reported 11 and are briefly described below.

The median lethal dose LD 50 of a venom was determined by i. In all experiments, the control groups of mice, regardless of whether 5x, 2. The neutralization potency P of the antiserum, defined as the amount of venom completely neutralized per unit volume of antiserum, was expressed as previously described Chippaux, J.

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Acta Tropica , Download references. This study was funded by a research grant no. IM to KR from the Chulabhorn Research Institute and a research grant from the Ministry of Higher Education, Government of Malaysia grant no. The authors are deeply grateful to Professor Arnold E. Ruoho and Dr.

Nicholas V. Cozzi of University of Wisconsin, Madison; Dr. James Dubbs and Ms Sukanya Earsakul of Chulabhorn Research Institute, and Professor Jirundon Yuvaniyama of Mahidol University and Dr.

Janeyuth Chaisakul for the valuable suggestions and assistance. Department of Microbiology, Faculty of Science, Mahidol University, Rama 6 Road, Bangkok, , Thailand.

Laboratory of Immunology, Chulabhorn Research Institute, Bangkok, , Thailand. Department of Molecular Medicine, Faculty of Medicine, University of Malaya, Kuala Lumpur, , Malaysia. Department of Microbiology, Faculty of Medicine, Chiang Mai University, Chiang Mai, , Thailand.

Veterinary Hospital, The Veterinary and Remount Department, The Royal Thai Army, Nakorn Pathom, , Thailand. Instituto Clodomiro Picado, Facultad de Microbiología, Universidad de Costa Rica, San José, Costa Rica.

Department of Pharmacology, Faculty of Medicine, University of Malaya, Kuala Lumpur, , Malaysia. You can also search for this author in PubMed Google Scholar.

planned the experiments; K. analyzed the data; K. Correspondence to Kavi Ratanabanangkoon or Choo Hock Tan. Open Access This article is licensed under a Creative Commons Attribution 4. Reprints and permissions. A pan-specific antiserum produced by a novel immunization strategy shows a high spectrum of neutralization against neurotoxic snake venoms.

Sci Rep 10 , Download citation. Received : 30 November Accepted : 30 April Published : 09 July Anyone you share the following link with will be able to read this content:.

Sorry, a shareable link is not currently available for this article. Provided by the Springer Nature SharedIt content-sharing initiative. International Journal of Peptide Research and Therapeutics This process of obtaining antivenom revolutionized the treatment of snakebite envenomation and influenced researchers worldwide.

Over more than years, production of antivenom, which must be tailored to species of snakes, remains much the same. Most antivenoms are produced in horses, some in sheep; a small amount of venom is injected into the animal, causing an immune system reaction and release of antibodies, which are later harvested via bleeding.

This blood plasma is then concentrated and purified into pharmaceutical grade antivenom. While the basic production method has remained little changed, many technological advances and purification processes have been introduced to achieve higher quality products and reduce side effects.

Additionally, in the s, antivenom began to be administered via the intravenous route injected into the vein as opposed to the subcutaneous route injected under the skin or intramuscular route injected into the muscle.

This has helped decrease severe reactions. By the end of the 20th century, antivenom manufacturers began to dwindle worldwide, due to complexity of production, high production expenses, and lack of a lucrative market. This has resulted in a dramatic increase in the price of some products over the last two decades.

Antivenom availability has also declined significantly. Meanwhile untested, unethically produced, or fake products have entered the market. To view Adobe PDF files, download current, free accessible plug-ins from Adobe's website.

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