Design, synthesis, and evaluation of Bothrops venom serine protease peptidic inhibitors

Abstract

Background: In Central and South America, snakebite envenomation is mainly caused by Bothrops spp. snakes, whose venoms feature significant biochemical richness, including serine proteases. The available bothropic antivenoms are efficient in avoiding fatalities, but do not completely neutralize venom serine proteases, which are co-responsible for some disorders observed during envenomation. Methods: In order to search for tools to improve the antivenom’s, 6-mer peptides were designed based on a specific substrate for Bothrops jararaca venom serine proteases, and then synthesized, with the intention to selectively inhibit these enzymes. Results: Using batroxobin as a snake venom serine protease model, two structurally similar inhibitor peptides were identified. When tested on B. jararaca venom, one of the new inhibitors displayed a good potential to inhibit the activity of the venom serine proteases. These inhibitors do not affect human serine proteases as human factor Xa and thrombin, due to their selectivity. Conclusion: Our study identified two small peptides able to inhibit bothropic serine proteases, but not human ones, can be used as tools to enhance knowledge of the venom composition and function. Moreover, one promising peptide (pepC) was identified that can be explored in the search for improving Bothrops spp. envenomation treatment.

Use of snake antivenom in the Region of the Americas: a systematic review

“… 65 Restoring coagulation is crucial to prevent bleeding, which could potentially lead to lethal complications. 43 Therefore, enhancing antivenom potency, by boosting the concentration of specific antibodies, and using enzyme inhibitors 66 , 67 may improve outcomes. We chose to report on measurements of coagulation only after 6 hours following antivenom therapy, to ensure that the outcome was attributable to the initial treatment.…”

Use of snake antivenom in the Region of the Americas: a systematic review

“… 65 Restoring coagulation is crucial to prevent bleeding, which could potentially lead to lethal complications. 43 Therefore, enhancing antivenom potency, by boosting the concentration of specific antibodies, and using enzyme inhibitors 66 , 67 may improve outcomes. We chose to report on measurements of coagulation only after 6 hours following antivenom therapy, to ensure that the outcome was attributable to the initial treatment.…”

Effect of Seaweed-Derived Fucoidans from Undaria pinnatifida and Fucus vesiculosus on Coagulant, Proteolytic, and Phospholipase A2 Activities of Snake Bothrops jararaca, B. jararacussu, and B. neuwiedi Venom

“…However, mAbs failed to neutralize the toxins of B. alternatus and B. neuwiedi venoms [ 41 ]. Da Silva et al [ 49 ] designed two synthetic peptides that inhibited SVSPs of B. jararaca crude venom and the purified serine protease Batroxobin, but not human trypsin, which is a serine protease. Undoubtedly, such peptides are a promising therapeutic tool for improving the treatment of SBE caused by Bothrops species.…”

In vitro immunoreactivity and in vivo neutralization of Trimeresurus gracilis venom with antivenoms targeting four pit viper species

“…Sousa et al [ 39 ] compared the composition and reactivity with multispecific Bothrops antivenom (SAB) of venoms collected from six species of Bothrops , Bothropoides and Rhinocerophis snakes, and found that P-III SVMPs are the most antigenic toxins in the venoms of snakes from the Bothrops complex, while class P-I SVMPs, SVSPs and PLA 2 s reacted with SAB in lower levels. Silva et al [ 40 , 41 ] discovered that the antivenom targeting Bothrops venom has limitations in addressing local effects such as hemorrhage and neutralizing SVSPs. However, they observed that supplementing the antivenom with a serine protease inhibitor can enhance its overall efficacy.…”

Use of snake antivenom in the Region of the Americas: a systematic review

“… 65 Restoring coagulation is crucial to prevent bleeding, which could potentially lead to lethal complications. 43 Therefore, enhancing antivenom potency, by boosting the concentration of specific antibodies, and using enzyme inhibitors 66 , 67 may improve outcomes. We chose to report on measurements of coagulation only after 6 hours following antivenom therapy, to ensure that the outcome was attributable to the initial treatment.…”

Effect of Seaweed-Derived Fucoidans from Undaria pinnatifida and Fucus vesiculosus on Coagulant, Proteolytic, and Phospholipase A2 Activities of Snake Bothrops jararaca, B. jararacussu, and B. neuwiedi Venom

“…However, mAbs failed to neutralize the toxins of B. alternatus and B. neuwiedi venoms [ 41 ]. Da Silva et al [ 49 ] designed two synthetic peptides that inhibited SVSPs of B. jararaca crude venom and the purified serine protease Batroxobin, but not human trypsin, which is a serine protease. Undoubtedly, such peptides are a promising therapeutic tool for improving the treatment of SBE caused by Bothrops species.…”

In vitro immunoreactivity and in vivo neutralization of Trimeresurus gracilis venom with antivenoms targeting four pit viper species

“…Sousa et al [ 39 ] compared the composition and reactivity with multispecific Bothrops antivenom (SAB) of venoms collected from six species of Bothrops , Bothropoides and Rhinocerophis snakes, and found that P-III SVMPs are the most antigenic toxins in the venoms of snakes from the Bothrops complex, while class P-I SVMPs, SVSPs and PLA 2 s reacted with SAB in lower levels. Silva et al [ 40 , 41 ] discovered that the antivenom targeting Bothrops venom has limitations in addressing local effects such as hemorrhage and neutralizing SVSPs. However, they observed that supplementing the antivenom with a serine protease inhibitor can enhance its overall efficacy.…”

Use of snake antivenom in the Region of the Americas: a systematic review

“… 65 Restoring coagulation is crucial to prevent bleeding, which could potentially lead to lethal complications. 43 Therefore, enhancing antivenom potency, by boosting the concentration of specific antibodies, and using enzyme inhibitors 66 , 67 may improve outcomes. We chose to report on measurements of coagulation only after 6 hours following antivenom therapy, to ensure that the outcome was attributable to the initial treatment.…”

Effect of Seaweed-Derived Fucoidans from Undaria pinnatifida and Fucus vesiculosus on Coagulant, Proteolytic, and Phospholipase A2 Activities of Snake Bothrops jararaca, B. jararacussu, and B. neuwiedi Venom

“…However, mAbs failed to neutralize the toxins of B. alternatus and B. neuwiedi venoms [ 41 ]. Da Silva et al [ 49 ] designed two synthetic peptides that inhibited SVSPs of B. jararaca crude venom and the purified serine protease Batroxobin, but not human trypsin, which is a serine protease. Undoubtedly, such peptides are a promising therapeutic tool for improving the treatment of SBE caused by Bothrops species.…”

In vitro immunoreactivity and in vivo neutralization of Trimeresurus gracilis venom with antivenoms targeting four pit viper species

“…Sousa et al [ 39 ] compared the composition and reactivity with multispecific Bothrops antivenom (SAB) of venoms collected from six species of Bothrops , Bothropoides and Rhinocerophis snakes, and found that P-III SVMPs are the most antigenic toxins in the venoms of snakes from the Bothrops complex, while class P-I SVMPs, SVSPs and PLA 2 s reacted with SAB in lower levels. Silva et al [ 40 , 41 ] discovered that the antivenom targeting Bothrops venom has limitations in addressing local effects such as hemorrhage and neutralizing SVSPs. However, they observed that supplementing the antivenom with a serine protease inhibitor can enhance its overall efficacy.…”