Snakebite envenomation (SBE) is an important public health challenge worldwide, being included in the list of neglected tropical diseases by the World Health Organization (WHO). It is estimated to affect close to 2.7 million people globally every year, especially in rural areas of tropical and subtropical countries (Binorkar and Jani, 2012; Feitosa et al., 2015; Adrião et al., 2022). According to an estimate of WHO, around 100,000 people annually die and 400,000 suffer disability due to SBE (Scheneider et al., 2021a; Liaqat et al., 2022).

Most snakebite cases occur in Africa, Asia, and Latin America. Brazil, with its large forest areas and great variety of venomous snakes, is among the countries with the highest risk of SBE in South America, with about 26,000–29,000 cases per year, followed by Venezuela (7,000), Colombia (3,000), Ecuador (1,500), Peru (1,500), and Bolivia (1,000) (Roriz et al., 2018; Scheneider et al., 2021a, 2021b).

Species of the genus Bothrops are responsible for 90% of the SBE in Latin America. Most of the Brazilian snakebite accident notifications are related to specimens of the Viperidae family, which comprise the genera Bothrops (known as ‘jararaca’) (83.3%), followed by Crotalus (‘cascavel’) (8.5%), and Lachesis (‘surucucu-pico-de-jaca’) (3.4%). In the Elapidae family, the genus Micrurus (true ‘coral’) is responsible for 0.8% of these accidents (Feitosa et al., 2015; Farias et al., 2018; Scheneider et al., 2021a; Konrath et al., 2022). Clinical manifestations of bothropic accidents are characterized by prominent local and systemic effects in the victims, such as inflammation, edema, hemorrhage, renal or cardiac failure, muscle necrosis, and eventually death (Otero et al., 2000; Jorge et al., 2017).

Bothrops jararaca (Wied-Neuwied, 1824) is the species that causes the greatest number of snakebites in South and Southeastern Brazil, around 60% in areas at sea level as well as over 1000 m altitude, or even in large cities (Moura et al., 2015; Machado et al., 2016; Jorge et al., 2017; Farias et al., 2018). The venom of B. jararaca is a mixture of active protein and peptides, including snake venom metalloproteinases (SVMPs), snake venom serine proteinases (SVSPs), phospholipases A2 (PLA2), L-amino acid oxidases (LAAOs), hyaluronidases, and bradykinin potentiators among others, and some of them are responsible for the most toxic activities (Kisaki et al., 2021).

The treatment recommended by WHO and the Ministry of Health of Brazil is performed by the intravenous injection of commercial antivenoms that are produced by hyperimmunization of equines. Antivenom has been available for 120 years and is highly effective in preventing mortality, especially if administered early in an adequate dose (Estevao-Costa et al., 2016). However, the utilization of antivenoms to treat morbidity and disability as consequences of snakebites has been limited by multiple factors, including adverse effects profile (fever or anaphylactic shock), stringent storage conditions, specificity issues, risk of immunological reactions, and high cost of production (Félix-Silva et al., 2017; Vásquez et al., 2013; Liaqat et al., 2022).

The inefficacy of antivenoms to neutralize the local effect of SBE may have serious consequences for recovery of victims, affecting the quality of life. Thus, seeking alternative treatments for SBE deserves deeper investigation. Over the years, many attempts have been made to find snake venom's antagonists from plants, since these organisms produce many compounds with unique and intriguing chemical structure. Some of these compounds may be useful as supplement or alternative treatments of SBE, and with less adverse events, better tolerability, and safety profile (Binorkar and Jani, 2012; Félix-Silva et al., 2017). The phytochemical analysis of various plants has revealed that phenols, alkaloids, triterpenoids, and steroids possess promising effects against the toxic activities caused by snake venom (Janardhan et al., 2014; Singh et al., 2017; Liaqat et al., 2022).

Currently, modern mass spectrometry techniques are widely employed for the identification and structural analysis of compounds in complex mixtures (Popov et al., 2022). In parallel, Computer-Aided Drug Design has emerged as a pivotal method for integrating the intricate complexities of biological systems with the predictive power of computational algorithms (Khan et al., 2020; Mostofa et al., 2023; Niazi and Mariam, 2024). Utilizing this data, fingerprint analysis can forecast which metabolites are likely contributors to specific biological activities even before isolating the target compounds. This approach complements experimental analysis and aids in the rational design of novel therapeutic agents (Darwish et al., 2022; Sweilam et al., 2022).

Siparuna species, Siparunaceae family, thrive in the tropical and subtropical regions of the Southern hemisphere. These plants hold significance in folk and traditional medicine due to their therapeutic properties, as evidenced by documented use in treating various illnesses (Leitão et al., 1999, 2000; Silva et al., 2021, Silva et al., 2021). In remote areas where access to prompt medical care is limited, patients often experience exacerbated conditions, leading to fatalities. Such circumstances prompt at-risk populations to develop community-based solutions to address their healthcare needs. For instance, in indigenous communities in Colombia, macerated leaves of Siparuna gesnerioides (Kunth) A.DC. are utilized to combat SBE (Vásquez et al., 2013, 2015; Félix-Silva et al., 2017), while decoctions or macerations of leaves, branches, and stems of Siparuna thecaphora (Poepp. & Endl.) A.DC. are employed in the form of drinks, external baths, and poultices to prevent SBE-related hemorrhaging (Otero et al., 2000; Gomes et al., 2010; Binorkar and Jani, 2012). Moreover, marginalized communities in the Amazon region of Brazil commonly rely on Siparuna apiosyce (Mart.) A.DC., (synonym: Siparuna brasiliensis), Siparuna guianensis Aublet and Siparuna ficoides S. S. Renner and Hausner, locally known as ‘Limão bravo’, ‘Negramina’, and ‘Yekuna’ respectively, in traditional medicine to mitigate the effects of snake bites (Leitão et al., 2000; Renner and Hausner, 2005; Rieder, 2013).

Based on these reports of Siparuna species against snakebite, and given the lack of identification of compounds in these plants responsible for such antivenom effect, this study aimed to investigate the correlation between the chemical compounds present in the extracts of Siparuna ficoides Renner & Hausner (originally identified as S. cristata Poepp. & Endl. A.DC.), Siparuna decipiens (Tul.) A.DC., Siparuna glycycarpa (Ducke) S.S. Renner & Hausner, Siparuna reginae (Tul.) A. DC., and Siparuna cymosa Tolm (originally identified as S. sarmentosa Perkins) (INPA, 2024), as potential candidates against the proteolytic and plasma coagulant by ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS), and multivariate statistics. Furthermore, it was also aimed to evaluate the potential of the extracts in inhibiting PLA2 activity, allowing inference of possible mechanisms of action of the extracts.