scholarly journals Neutralizing effects of small molecule inhibitors and metal chelators on coagulopathic Viperinae snake venom toxins

2020 ◽  
Author(s):  
Chunfang Xie ◽  
Laura Albulescu ◽  
Matyas A Bittenbinder ◽  
Govert Somsen ◽  
Freek Vonk ◽  
...  
Keyword(s):  
Small Molecule ◽  
Snake Venom ◽  
Small Molecule Inhibitors ◽  
Drug Repurposing ◽  
Phospholipase A ◽  
Proteomics Data ◽  
Snake Venoms ◽  
Analytical Technique ◽  
Venom Toxins ◽  
Snake Venom Metalloproteinase

AbstractAnimal-derived antivenoms are the only specific therapies currently available for the treatment of snake envenoming, but these products have a number of limitations associated with their efficacy, safety and affordability for use in tropical snakebite victims. Small molecule drugs and drug candidates are regarded as promising alternatives for filling the critical therapeutic gap between snake envenoming and effective treatment. In this study, by using an advanced analytical technique that combines chromatography, mass spectrometry and bioassaying, we investigated the effect of several small molecule inhibitors that target phospholipase A2 (varespladib) and snake venom metalloproteinase (marimastat, dimercaprol and DMPS) toxin families on inhibiting the activities of coagulopathic toxins found in Viperinae snake venoms. The venoms of Echis carinatus, Echis ocellatus, Daboia russelii and Bitis arietans, which are known for their potent coagulopathic toxicities, were fractionated in high resolution onto 384-well plates using liquid chromatography followed by coagulopathic bioassaying of the obtained fractions. Bioassay activities were correlated to parallel recorded mass spectrometric and proteomics data to assign the venom toxins responsible for coagulopathic activity and assess which of these toxins could be neutralized by the inhibitors under investigation. Our results showed that the phospholipase A2-inhibitor varespladib neutralized the vast majority of anticoagulation activities found across all of the tested snake venoms. Of the snake venom metalloproteinase inhibitors, marimastat demonstrated impressive neutralization of the procoagulation activities detected in all of the tested venoms, whereas dimercaprol and DMPS could only partially neutralize these activities at the doses tested. Our results provide additional support for the concept that combination of small molecules, particularly the combination of varespladib with marimastat, serve as a drug-repurposing opportunity to develop new broad-spectrum inhibitor-based therapies for snakebite envenoming.

scholarly journals Neutralizing Effects of Small Molecule Inhibitors and Metal Chelators on Coagulopathic Viperinae Snake Venom Toxins

Biomedicines ◽  
2020 ◽  
Vol 8 (9) ◽  
pp. 297 ◽  
Author(s):  
Chunfang Xie ◽  
Laura-Oana Albulescu ◽  
Mátyás A. Bittenbinder ◽  
Govert W. Somsen ◽  
Freek J. Vonk ◽  
...  
Keyword(s):  
Phospholipase A2 ◽  
Small Molecule ◽  
Snake Venom ◽  
Small Molecule Inhibitors ◽  
Drug Repurposing ◽  
Proteomics Data ◽  
Snake Venoms ◽  
Analytical Technique ◽  
Venom Toxins ◽  
Snake Venom Metalloproteinase

Animal-derived antivenoms are the only specific therapies currently available for the treatment of snake envenoming, but these products have a number of limitations associated with their efficacy, safety and affordability for use in tropical snakebite victims. Small molecule drugs and drug candidates are regarded as promising alternatives for filling the critical therapeutic gap between snake envenoming and effective treatment. In this study, by using an advanced analytical technique that combines chromatography, mass spectrometry and bioassaying, we investigated the effect of several small molecule inhibitors that target phospholipase A2 (varespladib) and snake venom metalloproteinase (marimastat, dimercaprol and DMPS) toxin families on inhibiting the activities of coagulopathic toxins found in Viperinae snake venoms. The venoms of Echis carinatus, Echis ocellatus, Daboia russelii and Bitis arietans, which are known for their potent haemotoxicities, were fractionated in high resolution onto 384-well plates using liquid chromatography followed by coagulopathic bioassaying of the obtained fractions. Bioassay activities were correlated to parallel recorded mass spectrometric and proteomics data to assign the venom toxins responsible for coagulopathic activity and assess which of these toxins could be neutralized by the inhibitors under investigation. Our results showed that the phospholipase A2-inhibitor varespladib neutralized the vast majority of anticoagulation activities found across all of the tested snake venoms. Of the snake venom metalloproteinase inhibitors, marimastat demonstrated impressive neutralization of the procoagulation activities detected in all of the tested venoms, whereas dimercaprol and DMPS could only partially neutralize these activities at the doses tested. Our results provide additional support for the concept that combinations of small molecules, particularly the combination of varespladib with marimastat, serve as a drug-repurposing opportunity to develop new broad-spectrum inhibitor-based therapies for snakebite envenoming.

scholarly journals Varespladib Inhibits the Phospholipase A2 and Coagulopathic Activities of Venom Components from Hemotoxic Snakes

Biomedicines ◽  
2020 ◽  
Vol 8 (6) ◽  
pp. 165 ◽  
Author(s):  
Chunfang Xie ◽  
Laura-Oana Albulescu ◽  
Kristina B. M. Still ◽  
Julien Slagboom ◽  
Yumei Zhao ◽  
...  
Keyword(s):  
Phospholipase A2 ◽  
Snake Venom ◽  
Snake Venoms ◽  
Venom Toxins ◽  
Inhibitory Molecule ◽  
Coagulation Assay ◽  
Potential Clinical Utility ◽  
Bothrops Asper ◽  
Echis Carinatus ◽  
Deinagkistrodon Acutus

Phospholipase A2 (PLA2) enzymes are important toxins found in many snake venoms, and they can exhibit a variety of toxic activities including causing hemolysis and/or anticoagulation. In this study, the inhibiting effects of the small molecule PLA2 inhibitor varespladib on snake venom PLA2s was investigated by nanofractionation analytics, which combined chromatography, mass spectrometry (MS), and bioassays. The venoms of the medically important snake species Bothrops asper, Calloselasma rhodostoma, Deinagkistrodon acutus, Daboia russelii, Echis carinatus, Echis ocellatus, and Oxyuranus scutellatus were separated by liquid chromatography (LC) followed by nanofractionation and interrogation of the fractions by a coagulation assay and a PLA2 assay. Next, we assessed the ability of varespladib to inhibit the activity of enzymatic PLA2s and the coagulopathic toxicities induced by fractionated snake venom toxins, and identified these bioactive venom toxins and those inhibited by varespladib by using parallel recorded LC-MS data and proteomics analysis. We demonstrated here that varespladib was not only capable of inhibiting the PLA2 activities of hemotoxic snake venoms, but can also effectively neutralize the coagulopathic toxicities (most profoundly anticoagulation) induced by venom toxins. While varespladib effectively inhibited PLA2 toxins responsible for anticoagulant effects, we also found some evidence that this inhibitory molecule can partially abrogate procoagulant venom effects caused by different toxin families. These findings further emphasize the potential clinical utility of varespladib in mitigating the toxic effects of certain snakebites.

scholarly journals Antivenom Neutralization of Coagulopathic Snake Venom Toxins Assessed by Bioactivity Profiling Using Nanofractionation Analytics

Toxins ◽  
2020 ◽  
Vol 12 (1) ◽  
pp. 53 ◽  
Author(s):  
Chunfang Xie ◽  
Julien Slagboom ◽  
Laura-Oana Albulescu ◽  
Ben Bruyneel ◽  
Kristina B. M. Still ◽  
...  
Keyword(s):  
Snake Venom ◽  
Serine Proteases ◽  
Clinical Symptoms ◽  
Neglected Tropical Diseases ◽  
Anticoagulant Activity ◽  
Proteomics Data ◽  
Neutralization Capacity ◽  
Venom Toxins ◽  
Coagulation Assay ◽  
Deinagkistrodon Acutus

Venomous snakebite is one of the world’s most lethal neglected tropical diseases. Animal-derived antivenoms are the only standardized specific therapies currently available for treating snakebite envenoming, but due to venom variation, often this treatment is not effective in counteracting all clinical symptoms caused by the multitude of injected toxins. In this study, the coagulopathic toxicities of venoms from the medically relevant snake species Bothrops asper, Calloselasma rhodostoma, Deinagkistrodon acutus, Daboia russelii, Echis carinatus and Echis ocellatus were assessed. The venoms were separated by liquid chromatography (LC) followed by nanofractionation and parallel mass spectrometry (MS). A recently developed high-throughput coagulation assay was employed to assess both the pro- and anticoagulant activity of separated venom toxins. The neutralization capacity of antivenoms on separated venom components was assessed and the coagulopathic venom peptides and enzymes that were either neutralized or remained active in the presence of antivenom were identified by correlating bioassay results with the MS data and with off-line generated proteomics data. The results showed that most snake venoms analyzed contained both procoagulants and anticoagulants. Most anticoagulants were identified as phospholipases A2s (PLA2s) and most procoagulants correlated with snake venom metalloproteinases (SVMPs) and serine proteases (SVSPs). This information can be used to better understand antivenom neutralization and can aid in the development of next-generation antivenom treatments.

scholarly journals Identification of small-molecule inhibitors of Zika virus infection and induced neural cell death via a drug repurposing screen

2016 ◽  
Vol 22 (10) ◽  
pp. 1101-1107 ◽  
Author(s):  
Miao Xu ◽  
Emily M Lee ◽  
Zhexing Wen ◽  
Yichen Cheng ◽  
Wei-Kai Huang ◽  
...  
Keyword(s):  
Cell Death ◽  
Virus Infection ◽  
Small Molecule ◽  
Zika Virus ◽  
Small Molecule Inhibitors ◽  
Drug Repurposing ◽  
Neural Cell ◽  
Zika Virus Infection

scholarly journals Carbon monoxide attenuates the effects of snake venoms containing metalloproteinases with fibrinogenase or thrombin-like activity on plasmatic coagulation

MedChemComm ◽  
2016 ◽  
Vol 7 (10) ◽  
pp. 1973-1979 ◽  
Author(s):  
Vance G. Nielsen ◽  
Charles M. Bazzell
Keyword(s):  
Carbon Monoxide ◽  
Human Plasma ◽  
Snake Venom ◽  
Plasma Velocity ◽  
Snake Venoms ◽  
Crotalus Atrox ◽  
Snake Venom Metalloproteinase ◽  
Plasmatic Coagulation

Carbon monoxide released from CORM-2 inhibitsCrotalus atroxsnake venom metalloproteinase mediated decreases in human plasma velocity of coagulation.

scholarly journals Anticoagulant Activity of Naja nigricollis Venom Is Mediated by Phospholipase A2 Toxins and Inhibited by Varespladib

Toxins ◽  
2021 ◽  
Vol 13 (5) ◽  
pp. 302
Author(s):  
Taline D. Kazandjian ◽  
Arif Arrahman ◽  
Kristina B. M. Still ◽  
Govert W. Somsen ◽  
Freek J. Vonk ◽  
...  
Keyword(s):  
Phospholipase A2 ◽  
Anticoagulant Activity ◽  
West African ◽  
Snake Venoms ◽  
The West ◽  
Coagulation Assays ◽  
Venom Toxins ◽  
African Black ◽  
Snake Venom Metalloproteinase ◽  
Species Specific

Bites from elapid snakes typically result in neurotoxic symptoms in snakebite victims. Neurotoxins are, therefore, often the focus of research relating to understanding the pathogenesis of elapid bites. However, recent evidence suggests that some elapid snake venoms contain anticoagulant toxins which may help neurotoxic components spread more rapidly. This study examines the effects of venom from the West African black-necked spitting cobra (Naja nigricollis) on blood coagulation and identifies potential coagulopathic toxins. An integrated RPLC-MS methodology, coupled with nanofractionation, was first used to separate venom components, followed by MS, proteomics and coagulopathic bioassays. Coagulation assays were performed on both crude and nanofractionated N. nigricollis venom toxins as well as PLA2s and 3FTx purified from the venom. Assays were then repeated with the addition of either the phospholipase A2 inhibitor varespladib or the snake venom metalloproteinase inhibitor marimastat to assess whether either toxin inhibitor is capable of neutralizing coagulopathic venom activity. Subsequent proteomic analysis was performed on nanofractionated bioactive venom toxins using tryptic digestion followed by nanoLC-MS/MS measurements, which were then identified using Swiss-Prot and species-specific database searches. Varespladib, but not marimastat, was found to significantly reduce the anticoagulant activity of N. nigricollis venom and MS and proteomics analyses confirmed that the anticoagulant venom components mostly consisted of PLA2 proteins. We, therefore, conclude that PLA2s are the most likely candidates responsible for anticoagulant effects stimulated by N. nigricollis venom.

scholarly journals Discovery of small molecule inhibitors for the snake venom metalloprotease BaP1 using in silico and in vitro tests

2017 ◽  
Vol 27 (9) ◽  
pp. 2018-2022
Author(s):  
Fabian Villalta-Romero ◽  
Luiz Borro ◽  
Boris Mandic ◽  
Teresa Escalante ◽  
Alexandra Rucavado ◽  
...  
Keyword(s):  
Small Molecule ◽  
Snake Venom ◽  
In Silico ◽  
Small Molecule Inhibitors ◽  
In Vitro Tests

scholarly journals Do Antibiotics Potentiate Proteases in Hemotoxic Snake Venoms?

Toxins ◽  
2020 ◽  
Vol 12 (4) ◽  
pp. 240
Author(s):  
Christoffer V. Sørensen ◽  
Cecilie Knudsen ◽  
Ulrich auf dem Keller ◽  
Konstantinos Kalogeropoulos ◽  
Cristina Gutiérrez-Jiménez ◽  
...  
Keyword(s):  
Snake Venom ◽  
Bacterial Infections ◽  
Enzymatic Activities ◽  
In Vitro Assays ◽  
Clinical Implications ◽  
Snake Venoms ◽  
Venom Toxins

Antibiotics are often administered with antivenom following snakebite envenomings in order to avoid secondary bacterial infections. However, to this date, no studies have evaluated whether antibiotics may have undesirable potentiating effects on snake venom. Herein, we demonstrate that four commonly used antibiotics affect the enzymatic activities of proteolytic snake venom toxins in two different in vitro assays. Similar findings in vivo could have clinical implications for snakebite management and require further examination.

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