Molecular docking and dynamics as a tool to study benzimidazole resistance in helminths: A scoping review

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Resistance against BZ drugs is due to mutations that change the amino acid comprising the β-tubulin protein, which negatively affects its interactions with BZ drug molecules. Several in silico modeling studies have been published to decipher the precise mechanism of BZ resistance, but inconsistencies on the resistance consequence mutations confer and the effect of different BZ ligands have led to further confusion regarding the exact mechanism of resistance. Hence, this scoping review was done to unravel the mechanism of BZ resistance based on published research on molecular docking and dynamics. Methods: A scoping review was conducted in ScienceDirect, MEDLINE via PubMed and Scopus using the search term “Benzimidazole Resistance AND Beta Tubulin AND Molecular Docking”. A total of 37 hits were recovered and from these 6 were included after selection, inclusion, and risk of bias assessment. Results: The six research papers included in this review studied several helminth species: Haemonchus conturtos, Trichinella spiralis, Ancylostoma duodenale, Ancylostoma caninum, Ancylostoma ceylanicum, Necator americanus, Trichuris trichiura, Trichuris suis, Anisakis simplex, Ascaris suum, Ascaridia galli, Parascaris equorum, Toxocara canis , and Fasciola hepatica . The benzimidazole resistance-associated mutations studied included F167Y (TTC, TTT → TAC, TAT), E198A (GAG, GAA → GCG, GCA), and F200Y (TTC, TTT → TAC, TAT). The results show that the E198A can markedly reduce the binding affinity of BZ ligand-β-tubulin interactions. The F167Y and F200Y also showed a similar effect that could vary based on the helminth species. The F200Y mutation can alter the conformation of the β-tubulin active site, negatively affecting drug binding. Conclusion: While the impact of these mutations can vary depending on the specific helminth species and the BZ drug involved, the overall findings highlight the importance of targeting these residues for the development of novel anthelmintic strategies to address emerging drug resistance. Parasitology Drug resistance computational biology docking helminths Figures Figure 1 Introduction Benzimidazole resistance continues to be an emerging grave concern in helminths of public and veterinary health concern. Benzimidazoles (BZ) are disruptors of microtubule polymerization by binding in the β subunit of the tubulin dimer (Whittaker et al., 2017 ). The binding of BZ drug molecules in the β-tubulin prevents the polymerization of tubulin subunits into microtubules, disrupting the formation of the cytoskeleton (Furtado et al., 2016 ). Benzimidazole drugs, like albendazole, mebendazole, and fenbendazole, are used for clinical treatment and preventive chemotherapy in humans and animals (TroCCAP, 2019 ; World Health Organization, 2011 ). Resistance against this drug class became a huge concern in the veterinary field as widespread reports of resistant livestock helminths, like Haemonchus c onturtos, Teladorsagia circumcincta , and Trichostrongylus colubriformis (Von Samson-Himmelstjerna et al., 2007 ). Among helminths of public health concern, soil-transmitted helminth infections that do not respond to conventional BZ treatment have been reported in several areas globally (Ng’etich et al., 2023 ; Schwenkenbecher et al., 2007 ). Recently, the emergence of BZ-resistant helminth infections among pets (e.g., canine hookworm) in the United States and Canada raises the zoonotic threat these treatment-irresponsive isolates pose (Jimenez Castro et al., 2021 ; Tenorio et al., 2024 ; Venkatesan et al., 2023 ). The resistance against BZ drugs is due to mutations that change the amino acid comprising the β-tubulin protein expressed by the helminth (Furtado et al., 2016 ). These amino acid substitutions are brought about by Single Nucleotide Polymorphisms (SNPs) (Von Samson-Himmelstjerna et al., 2007 ). These mutations include those that occur in amino acid positions 167 (Phenylalanine, F, TTC, TTT → Tyrosine, Y, TAC, TAT), 198 (Glutamic acid, E, GAG, GAA → Alanine, A, GCG, GCA) and 200 (Phenylalanine, F, TTC, TTT → Tyrosine, Y, TAC, TAT) (Furtado et al., 2016 ; Tenorio, 2023 ; Tenorio et al., 2024 ). These mutations alter the amino acid constitution of the expressed protein negatively affecting the binding of BZ drug molecules structurally or biochemically (Lacey and Gill, 1994 ). These mutations have been reported in a variety of worms that threaten humans and animals globally (Ng’etich et al., 2023 ). The atomic underpinnings of BZ resistance in helminths remain understudied, hence its precise mechanism has been put into question (Von Samson-Himmelstjerna et al., 2007 ). Several in silico modeling studies utilizing advances in computational biology have been undertaken to decipher the precise mechanism of BZ resistance. These research have included modeling the wild-type protein’s interaction with BZ drug ligands (Aguayo-Ortiz et al., 2013 ) and predicting the effects of BZ resistance mutations (Jones et al., 2022 ). However, inconsistency regarding the resistance effects each mutation confers and the consequences of utilizing numerous BZ derivatives as ligands has led to further confusion regarding the exact mechanism of resistance. Hence, this scoping review was done to unravel the mechanism of BZ resistance based on published research that used molecular docking and dynamics. The results show that the E198A can drive down the binding affinity of BZ ligand-β-tubulin interactions regardless of the species and drug used. The F167Y and F200Y also showed a similar effect that could vary based on the helminth species Materials and Methods Research Questions The scoping review was done based on the guidelines reported by the PRISMA-ScR (PRISMA Extension for Scoping Reviews) (Tricco et al., 2018) (https://www.prisma-statement.org/scoping). Based on published molecular docking and dynamics studies, this research aims to determine the mechanism of benzimidazole. Specifically, this research answers the following questions: 1. What are the in silico underpinnings of benzimidazole resistance based on molecular docking and dynamics study? 2. What are the consequences of these mutations on the measurement of binding efficiency of the β-tubulins-benzimidazole drug complex? 3. What are the consequences of these mutations on the interactions between the β-tubulins and the benzimidazole drug ligand/s? Search Strategy A systematic search was done in three research databases. Scopus (https://www.scopus.com/search), ScienceDirect (https://www.sciencedirect.com/), and MEDLINE via PubMed (https://pubmed.ncbi.nlm.nih.gov/) were searched using the search term “Benzimidazole Resistance AND Beta Tubulin AND Molecular Docking.” The literature search was done on 17 September 2024. The .ris file of the search results was downloaded. Study Selection, Strategy, and Eligibility Using the Mendeley citation manager (https://www.mendeley.com), the .ris files were uploaded and utilized for the selection and eligibility assessment. First, duplicates and records with no titles and abstracts (e.g., indexes) were removed. Second, an initial evaluation based on the title and abstract was done. Full-length articles of the studies were accessed for further eligibility appraisal. Figure 1 summarizes the systematic literature search, selection, and eligibility evaluation done. A study was considered eligible for selection if it fulfilled any of the following inclusion criteria: 1. Studies that utilized molecular docking in assessing the in silico effects of the BZ resistance mutations; 2. Studies that utilized molecular docking in assessing the in silico effects of the BZ resistance mutations From the included studies, papers that did not meet the following criteria were excluded: 1. Studies that did not report docking scoring functions (i.e., binding affinities) and/or binding free energies (e.g., MM-PBSA or MM-GBSA); 2. Studies that did not report the effects of BZ resistance mutations on the interaction between β-tubulins and benzimidazole drug ligand/s; 3. Studies done using non-helminth β-tubulins as macromolecules; 4. Studies that did not use commercially available benzimidazole drugs as ligands; and 5. Studies that utilized newly designed and synthesized benzimidazole derivatives. 6. Research not in the English language. Risk of Bias Assessment Due to the in silico and computational nature of the studies being reviewed, traditional checklists for laboratory experiments are not well-suited as the method of bias assessment. Hence, we developed a simple checklist that is based on the quality of the modeled β-tubulin macromolecule, ligand preparation, docking software, simulation quality, and data analysis utilized. The tool is in the form of a 13-item close-ended questionnaire (Supplementary Table 1). All included studies were evaluated using this tool. Data Acquisition and Synthesis The author’s name, year of publication, software used in molecular docking and/or system used in molecular dynamics, helminth species of the β-tubulins used as the macromolecule, benzimidazole ligand used, BZ resistance mutation evaluated, docking scoring functions and/or binding free energies of the complex, description of the changes in interactions, and relative resistance-associated effects were the data acquired from the selected studies. Simple descriptive statistics, like counts and frequencies, were used to describe and synthesize the results of this scoping review. Results Characteristics of the studies included A total of 37 hits were found in the three databases searched (Fig. 1 ). Sixteen of these were removed due to duplication. The full text of one article was not accessed. After the eligibility screening, two were removed for not reporting docking scoring function, eight were removed for not using helminth β-tubulins, three were dropped for using newly designed benzimidazole ligands, and one did not report the effects of the docked complexes. In total, six research papers were included in this scoping review. The six research papers included in this review studied several helminth species: Haemonchus conturtos, Trichinella spiralis, Ancylostoma duodenale, Ancylostoma caninum, Ancylostoma ceylanicum, Necator americanus, Trichuris trichiura, Trichuris suis, Anisakis simplex, Ascaris suum, Ascaridia galli, Parascaris equorum, Toxocara canis , and Fasciola hepatica. The benzimidazole resistance-associated mutations studied included F167Y (TTC, TTT → TAC, TAT), E198A (GAG, GAA → GCG, GCA), and F200Y (TTC, TTT → TAC, TAT). In silico effects of BZ resistance-associated mutations Table 1 summarizes computational studies investigating the molecular mechanisms underlying benzimidazole (BZ) drug resistance in various helminth species. The results highlight the crucial role of specific amino acid residues (e.g., F167, E198, F200) in BZ binding and the potential mechanisms through which mutations in these residues can confer resistance. Mutations in specific amino acid residues (F167, E198, F200) within β-tubulin proteins are frequently associated with BZ drug resistance. These mutations can disrupt the conformation of the β-tubulin active site, destabilize BZ binding, and reduce drug efficacy. Moreover, mutations, particularly at position 198, can lead to the loss of hydrogen bonding interactions between BZ drugs and β-tubulin, contributing to resistance. Further, mutations at positions 167 and 200 may interfere with the opening of the binding site or the internalization of BZ ligands. The impact of mutations on resistance can vary across different helminth species. For example, mutations in F167Y may have a greater impact on strongyle parasites compared to other soil-transmitted helminths. Overall, the results suggest that BZ resistance is primarily attributed to alterations in the β-tubulin binding site, hindering the effective interaction between BZ drugs and their target protein. Table 1 The key results of the studies included in this scoping review. STUDY CODE AUTHORS YEAR PUBLISHED MOLECULAR DOCKING SOFTWARE MOLECULAR DYNAMICS SYSTEM HELMINTH SPECIES β-TUBULIN EVALUATED BZ LIGAND USED DOCKING SCORING FUNCTION(Kcal/mol) BINDING FREE ENERGY (Kcal/mol) RESISTANCE EFFECTS NOTED S1 Aguayo-Ortiz et al. ( 2013 b) 2013 AutoDock 4.2 GROMACS 4.5.3 Haemonchus conturtos Wild-type Albendazole -8.10 -68.41 The mutated and unsusceptible β-tubulin models suggest that the primary cause of BZ resistance is likely due to an amino acid modification at position 198, resulting in the loss of hydrogen bonding interactions. Conversely, the substitution of phenylalanine for tyrosine at positions 167 and 200 implies that the inhibitory mechanism may occur either during the opening of the binding site or the internalization of the ligand. Carbendazim -6.98 -55.08 Oxibendazole -8.17 -67.27 Parbendazole -8.09 -65.59 Luxabendazole -9.46 -64.77 Mebendazole -9.43 -60.53 Nocodazole -9.36 -64.61 F167Y Albendazole -8.07 N/A Carbendazim -7.02 Oxibendazole -8.09 Parbendazole -8.15 Luxabendazole -9.26 Mebendazole -9.46 Nocodazole -9.35 E198A Albendazole -7.03 Carbendazim -6.37 Oxibendazole -6.66 Parbendazole -7.05 Luxabendazole -8.82 Mebendazole -8.26 Nocodazole -7.99 F200Y Albendazole -8.22 Carbendazim -7.19 Oxibendazole -8.28 Parbendazole -8.26 Luxabendazole -9.62 Mebendazole -9.48 Nocodazole -9.35 S2 Aguayo-Ortiz et al. ( 2013 a) 2013 AutoDock 4.2 GROMACS 4.5.3 Trichinella spiralis Wild-type Albendazole -7.70 -53.67 The modeling study found that the binding site for BZ aligns with previous experimental findings. This site includes amino acids linked to resistance mutations (F167Y, E198A, and F200Y), and overlaps with the colchicine-binding site. Molecular docking and dynamics calculations of BZ drugs reveal that they are stabilized within the binding site primarily through hydrogen bonds with specific amino acids (e.g., Thr165, Glu198, Cys239 and Gln134). Carbendazim -7.05 -21.74 Oxibendazole -7.41 -31.92 Parbendazole -7.93 -37.20 Luxabendazole -9.41 -44.71 Mebendazole -8.73 -43.04 Nocodazole -8.82 -34.34 S3 Jones et al. ( 2022 a) 2022 Autodock vina v1.1.2 Molecular Operating Environment (MOE) 2020.01 Ascaris suum Wild-type Albendazole -8.46 N/A In silico docking studies suggest that BZ drugs can bind to all Ascarid β-tubulin isotypes. Ascarid β-tubulin isotype A was further analyzed using molecular dynamics simulations. These simulations revealed the critical role of amino acid E198 in BZ-β-tubulin interactions. Mutations at E198A and F200Y were found to alter benzimidazole binding, while the F167Y mutation had no significant impact. F167Y Albendazole -8.04 E198A Albendazole -8.53 F200Y Albendazole -8.16 S4 Jones et al. ( 2022 b) 2022 Autodock vina v1.1.2 Molecular Operating Environment (MOE) version 2020.01 Ancylostoma duodenale Wild-type Albendazole -8.55 N/A The study indicates that BZ acts through similar mechanisms in various helminth species. The amino acid E198 plays a crucial role in BZ binding. However, the Q134–F167Y interaction observed in Ancylostoma caninum , along with the presence of the F167Y SNP in susceptible Ascaris populations and its reduced frequency after BZ treatment in Trichuris trichiura from previous studies, suggests that mutations in F167Y may have a greater impact on strongyle parasites compared to other STHs. F167Y Albendazole -8.19 E198A Albendazole -8.14 F200Y Albendazole -7.72 Trichuris trichiura Wild- Type Albendazole -8.53 F167Y Albendazole -9.82 E198A Albendazole -7.78 F200Y Albendazole -10.48 Anisakis simplex Wild-type Albendazole -7.74 Ascaridia galli Wild-type Albendazole -8.25 Parascaris equorum Wild-type Albendazole -8.19 Toxocara canis Wild-type Albendazole -8.82 Ancylostoma caninum Wild-type Albendazole -7.94 Ancylostoma ceylanicum Wild-type Albendazole -8.29 Necator americanus Wild-type Albendazole -7.54 Trichuris suis Wild-type Albendazole -8.53 S5 Olivares-Ferretti et al. (2023) 2023 AutoDock Vina program in PyRx software N/A Fasciola hepatica Wild-type isotype 1 Triclabendazole -6.87 N/A The nucleotide binding site on F. hepatica β-tubulin exhibits a higher affinity for ligands than other known binding sites, such as colchicine, albendazole, the T7 loop, and pβVII. Ligand binding to the polymerization site of β-tubulin can disrupt microtubule formation. Triclabendazole demonstrated significantly higher binding affinity than other ligands across all β-tubulin isotypes. Computational analysis has provided new insights into the mechanism of action of triclabendazole on F. hepatica β-tubulin. Wild-type isotype 2 Triclabendazole -6.40 Wild-type isotype 3 Triclabendazole -6.38 Wild-type isotype 4 Triclabendazole -6.33 Wild-type isotype 5 Triclabendazole -6.17 Wild-type isotype 6 Triclabendazole -6.48 S6 Yashica et al. (2024) 2024 AutoDock Tools in MGL tools 1.5.6 N/A Haemonchus contortus Wild-type Albendazole -8.51 N/A The in silico study suggests that mutations at amino acid position 200 can disrupt the conformation of the H. conturtos β-tubulin active site and destabilize albendazole binding. Effects of Mutations on Docking Scoring Function and Binding Free Energy Based on the results of the included studies, the BZ resistance mutations often lead to a decrease in the docking scoring function and binding free energy, indicating a weaker interaction between the BZ drug and the mutated β-tubulin protein. However, the magnitude of the effect can vary depending on the specific mutation and the BZ drug involved. While F167Y mutations can sometimes decrease binding affinity, their effect is often less pronounced compared to mutations at other residues. Also, the impact of F167Y may vary across different helminth species. Meanwhile, the E198A mutation consistently leads to a marked decrease in binding affinity, suggesting a crucial role of this residue in BZ binding. The E198A mutation likely disrupts hydrogen bonding interactions between the BZ drug and β-tubulin. The F200Y mutations generally result in a moderate decrease in binding affinity. This mutation may alter the conformation of the β-tubulin active site, affecting drug binding. The combined effects of multiple mutations can further reduce binding affinity and enhance resistance. Likewise, the impact of mutations may vary depending on the specific BZ drug being considered. Overall, the results suggest that mutations in these key residues can disrupt the structural and electrostatic interactions between BZ drugs and β-tubulin, leading to reduced binding affinity and ultimately, drug resistance. Discussion This scoping review was conducted with the aim of unraveling the mechanism of BZ resistance based on published research on molecular docking and dynamics. MEDLINE via PubMed, Scopus, and Science Direct were searched. A total of six eligible studies were included, which encompassed research that utilized β-tubulins from several helminth species and numerous benzimidazole ligands. Of the BZ mutations studied, E198A showed that it can drive down the binding affinity of BZ ligand-β-tubulin interactions. The F167Y and F200Y also showed a similar effect that could vary based on the helminth species. The F200Y mutation can alter the conformation of the β-tubulin active site, negatively affecting drug binding. The studies included in this scoping review assessed both wild-type and mutated helminth β-tubulins. The three canonical BZ resistance mutations—F167Y, E198A, and F200Y—that are induced by SNPs were all evaluated (Furtado et al., 2016 ). These mutations have been reported in many helminth species of public and veterinary health concern from many parts of the world (Diawara et al., 2013 ; George et al., 2022 ; Jimenez Castro et al., 2021 ). Hence, their inclusion in computational studies was warranted. However, their actual contribution to conferring resistance has been put into question (Lacey and Gill, 1994 ). Also, several recent evidence of alternative resistance mechanisms, such as enzymatic biotransformation of drugs using UDP-glycosyltransferases and long non-coding RNA interference (Chen et al., 2024 ; Dimunová et al., 2022 ), have been reported. However, laboratory research using gene editing that encoded resistance-associated mutations in Caenorhabditis elegans showed their actual potential to confer resistance, as previously mentioned (Dilks et al., 2021 , 2020 ). Particularly, varying mutations that have been reported in amino acid position 198 have been reported to make edited C. elegans significantly more resistant to benzimidazole treatment compared to their wild-type counterpart (Dilks et al., 2021 ). This result echoes the finding in this systematic review that the E198A mutation had a consistent negative effect on the binding of BZ drugs and helminth β-tubulins indicating their important role in conferring resistance. This scoping review has several limitations. First, only a few accessible databases were searched, hence there might be other research not indexed in these databases that were missed. However, searching only known indexing databases, like MEDLINE via PubMed and Scopus, ensures that only quality research papers from reputable journals are included. Second, the in silico nature of the research targeted for this review may lead to conclusions of insufficient evidence for the conclusions that they present due to the lack of laboratory confirmation. However, advances in computational biology have assured that the predictions are of high quality and accuracy, particularly for molecular docking and dynamics studies (Santos et al., 2019 ; Singh et al., 2022 ). Conclusion The computational studies included in this scoping review provide valuable insights into the molecular mechanisms underlying BZ drug resistance in various helminth species. The results consistently demonstrate that mutations in specific amino acid residues within β-tubulin proteins, particularly F167, E198, and F200, play a critical role in conferring resistance. These mutations disrupt the structural and electrostatic interactions between BZ drugs and helminth β-tubulin, leading to reduced binding affinity and ultimately, drug resistance. While the impact of these mutations can vary depending on the specific helminth species and the BZ drug involved, the overall findings highlight the importance of targeting these residues for the development of novel anthelmintic strategies to address emerging drug resistance. Future studies could explore additional mutations, investigate the interactions between BZ drugs and other β-tubulin isoforms, or investigate the potential for combination therapies to overcome resistance. Declarations Supplementary Materials Supplementary tables and figures are available at [DOI}. Acknowledgments None. Funding No funding was received to assist with the preparation of this manuscript. Conflict of Interests The author declares that there is no conflict of any financial or non-financial interests regarding the publication of this paper. Ethics Approval Not applicable Consent to participate Not applicable Consent for publication Not applicable Data Availability Not applicable References Aguayo-Ortiz, R., Méndez-Lucio, O., Medina-Franco, J.L., Castillo, R., Yépez-Mulia, L., Hernández-Luis, F., Hernández-Campos, A., 2013. Towards the identification of the binding site of benzimidazoles to β-tubulin of Trichinella spiralis: Insights from computational and experimental data. Journal of Molecular Graphics and Modelling 41, 12–19. https://doi.org/10.1016/j.jmgm.2013.01.007 Chen, X., Wang, T., Guo, W., Yan, X., Kou, H., Yu, Y., Liu, C., Gao, W., Wang, W., Wang, R., 2024. Transcriptome reveals the roles and potential mechanisms of lncRNAs in the regulation of albendazole resistance in Haemonchus contortus. BMC Genomics 25, 188. https://doi.org/10.1186/s12864-024-10096-6 Diawara, A., Halpenny, C.M., Churcher, T.S., Mwandawiro, C., Kihara, J., Kaplan, R.M., Streit, T.G., Idaghdour, Y., Scott, M.E., Basáñez, M.-G., Prichard, R.K., 2013. Association between Response to Albendazole Treatment and β-Tubulin Genotype Frequencies in Soil-transmitted Helminths. PLoS Negl Trop Dis 7, e2247. https://doi.org/10.1371/journal.pntd.0002247 Dilks, C.M., Hahnel, S.R., Sheng, Q., Long, L., McGrath, P.T., Andersen, E.C., 2020. Quantitative benzimidazole resistance and fitness effects of parasitic nematode beta-tubulin alleles. Int J Parasitol Drugs Drug Resist 14, 28–36. https://doi.org/10.1016/j.ijpddr.2020.08.003 Dilks, C.M., Koury, E.J., Buchanan, C.M., Andersen, E.C., 2021. Newly identified parasitic nematode beta-tubulin alleles confer resistance to benzimidazoles. International Journal for Parasitology: Drugs and Drug Resistance 17, 168–175. https://doi.org/10.1016/j.ijpddr.2021.09.006 Dimunová, D., Navrátilová, M., Kellerová, P., Ambrož, M., Skálová, L., Matoušková, P., 2022. The induction and inhibition of UDP-glycosyltransferases in Haemonchus contortus and their role in the metabolism of albendazole. International Journal for Parasitology: Drugs and Drug Resistance 19, 56–64. https://doi.org/10.1016/j.ijpddr.2022.06.001 Furtado, L.F.V., De Paiva Bello, A.C.P., Rabelo, É.M.L., 2016. Benzimidazole resistance in helminths: From problem to diagnosis. Acta Tropica 162, 95–102. https://doi.org/10.1016/j.actatropica.2016.06.021 George, S., Suwondo, P., Akorli, J., Otchere, J., Harrison, L.M., Bilguvar, K., Knight, J.R., Humphries, D., Wilson, M.D., Caccone, A., Cappello, M., 2022. Application of multiplex amplicon deep-sequencing (MAD-seq) to screen for putative drug resistance markers in the Necator americanus isotype-1 β-tubulin gene. Sci Rep 12, 11459. https://doi.org/10.1038/s41598-022-15718-1 Jimenez Castro, P.D., Venkatesan, A., Redman, E., Chen, R., Malatesta, A., Huff, H., Zuluaga Salazar, D.A., Avramenko, R., Gilleard, J.S., Kaplan, R.M., 2021. Multiple drug resistance in hookworms infecting greyhound dogs in the USA. International Journal for Parasitology: Drugs and Drug Resistance 17, 107–117. https://doi.org/10.1016/j.ijpddr.2021.08.005 Jones, B.P., van Vliet, A.H.M., LaCourse, E.J., Betson, M., 2022. Identification of key interactions of benzimidazole resistance-associated amino acid mutations in Ascaris β-tubulins by molecular docking simulations. Sci Rep 12, 13725. https://doi.org/10.1038/s41598-022-16765-4 Lacey, E., Gill, J.H., 1994. Biochemistry of benzimidazole resistance. Acta Trop 56, 245–262. https://doi.org/10.1016/0001-706x(94)90066-3 Ng’etich, A.I., Amoah, I.D., Bux, F., Kumari, S., 2023. Anthelmintic resistance in soil-transmitted helminths: One-Health considerations. Parasitol Res 123, 62. https://doi.org/10.1007/s00436-023-08088-8 Santos, L.H.S., Ferreira, R.S., Caffarena, E.R., 2019. Integrating Molecular Docking and Molecular Dynamics Simulations, in: De Azevedo, W.F. (Ed.), Docking Screens for Drug Discovery, Methods in Molecular Biology. Springer New York, New York, NY, pp. 13–34. https://doi.org/10.1007/978-1-4939-9752-7_2 Schwenkenbecher, J.M., Albonico, M., Bickle, Q., Kaplan, R.M., 2007. Characterization of beta-tubulin genes in hookworms and investigation of resistance-associated mutations using real-time PCR. Mol Biochem Parasitol 156, 167–174. https://doi.org/10.1016/j.molbiopara.2007.07.019 Singh, S., Bani Baker, Q., Singh, D.B., 2022. Molecular docking and molecular dynamics simulation, in: Bioinformatics. Elsevier, pp. 291–304. https://doi.org/10.1016/B978-0-323-89775-4.00014-6 Tenorio, J.C.B., 2023. Canine hookworms in the Philippines—Very common but very much neglected in veterinary research. Frontiers in Veterinary Science 10. Tenorio, J.C.B., Tabios, I.K.B., Inpankaew, T., Ybañez, A.P., Tiwananthagorn, S., Tangkawattana, S., Suttiprapa, S., 2024. Ancylostoma ceylanicum and other zoonotic canine hookworms: neglected public and animal health risks in the Asia–Pacific region. Animal Diseases 4, 11. https://doi.org/10.1186/s44149-024-00117-y Tricco, A.C., Lillie, E., Zarin, W., O’Brien, K.K., Colquhoun, H., Levac, D., Moher, D., Peters, M.D.J., Horsley, T., Weeks, L., Hempel, S., Akl, E.A., Chang, C., McGowan, J., Stewart, L., Hartling, L., Aldcroft, A., Wilson, M.G., Garritty, C., Lewin, S., Godfrey, C.M., Macdonald, M.T., Langlois, E.V., Soares-Weiser, K., Moriarty, J., Clifford, T., Tunçalp, Ö., Straus, S.E., 2018. PRISMA Extension for Scoping Reviews (PRISMA-ScR): Checklist and Explanation. Ann Intern Med 169, 467–473. https://doi.org/10.7326/M18-0850 TroCCAP, 2019. Guidelines for the diagnosis, treatment and control of canine endoparasites in the tropics, 2nd ed. Venkatesan, A., Castro, P.D.J., Morosetti, A., Horvath, H., Chen, R., Redman, E., Dunn, K., Collins, J.B., Fraser, J.S., Andersen, E.C., Kaplan, R.M., Gilleard, J.S., 2023. Molecular evidence of widespread benzimidazole drug resistance in Ancylostoma caninum from domestic dogs throughout the USA and discovery of a novel β-tubulin benzimidazole resistance mutation. PLOS Pathogens 19, e1011146. https://doi.org/10.1371/journal.ppat.1011146 Von Samson-Himmelstjerna, G., Blackhall, W.J., McCARTHY, J.S., Skuce, P.J., 2007. Single nucleotide polymorphism (SNP) markers for benzimidazole resistance in veterinary nematodes. Parasitology 134, 1077–1086. https://doi.org/10.1017/S0031182007000054 Whittaker, J.H., Carlson, S.A., Jones, D.E., Brewer, M.T., 2017. Molecular mechanisms for anthelmintic resistance in strongyle nematode parasites of veterinary importance. J. vet. Pharmacol. Therap. 40, 105–115. https://doi.org/10.1111/jvp.12330 World Health Organization, 2011. Helminth control in school-age children : a guide for managers of control programmes. World Health Organization. Additional Declarations The authors declare no competing interests. Supplementary Files SupplementaryMaterials.docx Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-5476123","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Systematic Review","associatedPublications":[],"authors":[{"id":379478264,"identity":"0d3daa54-23ef-4822-b210-5e101d75fb7f","order_by":0,"name":"Jan Clyden Tenorio","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA5UlEQVRIiWNgGAWjYBACxgYeCIONgfkAhAWkmAloYWyAaGFLIE4LAwNUC5BhQJwW5vazxx983GGTz8fe803yZw6DHN+NBLbHBfgc1pOX2DjzTJplG8/ZbdK82xiMJW8ksBvPwKdlBo9hM2/bYQM2idxt0ozbGBI3AG2R5iGk5S9YS84zyZ/bGOqJ08II0cImAXRYggFBLUC/zOxtSzNg4zlmbM27TcJw5pmH7cb4tBi2nz3w4WebjYF8e/PDmz+32cjzHU8+9hivlgZUvgTI5jY8GhgY5LEJsuHVMgpGwSgYBSMOAAAZ6UjSoAjy5gAAAABJRU5ErkJggg==","orcid":"","institution":"University of Southern Mindanao","correspondingAuthor":true,"prefix":"","firstName":"Jan","middleName":"Clyden","lastName":"Tenorio","suffix":""}],"badges":[],"createdAt":"2024-11-18 12:52:55","currentVersionCode":1,"declarations":{"humanSubjects":false,"vertebrateSubjects":false,"conflictsOfInterestStatement":false,"humanSubjectEthicalGuidelines":false,"humanSubjectConsent":false,"humanSubjectClinicalTrial":false,"humanSubjectCaseReport":false,"vertebrateSubjectEthicalGuidelines":false},"doi":"10.21203/rs.3.rs-5476123/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-5476123/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":69298711,"identity":"d36ec667-c02e-475a-b4be-4fe09c052d78","added_by":"auto","created_at":"2024-11-19 02:18:37","extension":"jpg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":624357,"visible":true,"origin":"","legend":"\u003cp\u003ePRISMA-ScR (Preferred Reporting Items for Systematic Reviews and Meta-Analyses-Extension for Scoping Reviews) flowchart of the screening, selection, and eligibility selection in this study.\u003c/p\u003e","description":"","filename":"Figure1.PRISMAFlowchart.jpg","url":"https://assets-eu.researchsquare.com/files/rs-5476123/v1/72d8556752908833f9428d56.jpg"},{"id":69299490,"identity":"1b9f4fe6-8d66-4094-b9ff-5c979a378252","added_by":"auto","created_at":"2024-11-19 02:26:38","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1239172,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-5476123/v1/47c08b2b-f108-4750-9e60-b252e24e3a0b.pdf"},{"id":69298712,"identity":"0cff8020-3b40-4852-8591-e6e32492f228","added_by":"auto","created_at":"2024-11-19 02:18:37","extension":"docx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":17981,"visible":true,"origin":"","legend":"","description":"","filename":"SupplementaryMaterials.docx","url":"https://assets-eu.researchsquare.com/files/rs-5476123/v1/80004a01623c3bd72b7cd79d.docx"}],"financialInterests":"The authors declare no competing interests.","formattedTitle":"\u003cp\u003e\u003cstrong\u003eMolecular docking and dynamics as a tool to study benzimidazole resistance in helminths: A scoping review\u003c/strong\u003e\u003c/p\u003e","fulltext":[{"header":"Introduction","content":"\u003cp\u003eBenzimidazole resistance continues to be an emerging grave concern in helminths of public and veterinary health concern. Benzimidazoles (BZ) are disruptors of microtubule polymerization by binding in the β subunit of the tubulin dimer (Whittaker et al., \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). The binding of BZ drug molecules in the β-tubulin prevents the polymerization of tubulin subunits into microtubules, disrupting the formation of the cytoskeleton (Furtado et al., \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). Benzimidazole drugs, like albendazole, mebendazole, and fenbendazole, are used for clinical treatment and preventive chemotherapy in humans and animals (TroCCAP, \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; World Health Organization, \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2011\u003c/span\u003e). Resistance against this drug class became a huge concern in the veterinary field as widespread reports of resistant livestock helminths, like \u003cem\u003eHaemonchus\u003c/em\u003e c\u003cem\u003eonturtos, Teladorsagia circumcincta\u003c/em\u003e, and \u003cem\u003eTrichostrongylus colubriformis\u003c/em\u003e (Von Samson-Himmelstjerna et al., \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2007\u003c/span\u003e). Among helminths of public health concern, soil-transmitted helminth infections that do not respond to conventional BZ treatment have been reported in several areas globally (Ng\u0026rsquo;etich et al., \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Schwenkenbecher et al., \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2007\u003c/span\u003e). Recently, the emergence of BZ-resistant helminth infections among pets (e.g., canine hookworm) in the United States and Canada raises the zoonotic threat these treatment-irresponsive isolates pose (Jimenez Castro et al., \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Tenorio et al., \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; Venkatesan et al., \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2023\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe resistance against BZ drugs is due to mutations that change the amino acid comprising the β-tubulin protein expressed by the helminth (Furtado et al., \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). These amino acid substitutions are brought about by Single Nucleotide Polymorphisms (SNPs) (Von Samson-Himmelstjerna et al., \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2007\u003c/span\u003e). These mutations include those that occur in amino acid positions 167 (Phenylalanine, F, TTC, TTT \u0026rarr; Tyrosine, Y, TAC, TAT), 198 (Glutamic acid, E, GAG, GAA \u0026rarr; Alanine, A, GCG, GCA) and 200 (Phenylalanine, F, TTC, TTT \u0026rarr; Tyrosine, Y, TAC, TAT) (Furtado et al., \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Tenorio, \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Tenorio et al., \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). These mutations alter the amino acid constitution of the expressed protein negatively affecting the binding of BZ drug molecules structurally or biochemically (Lacey and Gill, \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e1994\u003c/span\u003e). These mutations have been reported in a variety of worms that threaten humans and animals globally (Ng\u0026rsquo;etich et al., \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). The atomic underpinnings of BZ resistance in helminths remain understudied, hence its precise mechanism has been put into question (Von Samson-Himmelstjerna et al., \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2007\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eSeveral \u003cem\u003ein silico\u003c/em\u003e modeling studies utilizing advances in computational biology have been undertaken to decipher the precise mechanism of BZ resistance. These research have included modeling the wild-type protein\u0026rsquo;s interaction with BZ drug ligands (Aguayo-Ortiz et al., \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2013\u003c/span\u003e) and predicting the effects of BZ resistance mutations (Jones et al., \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). However, inconsistency regarding the resistance effects each mutation confers and the consequences of utilizing numerous BZ derivatives as ligands has led to further confusion regarding the exact mechanism of resistance. Hence, this scoping review was done to unravel the mechanism of BZ resistance based on published research that used molecular docking and dynamics. The results show that the E198A can drive down the binding affinity of BZ ligand-β-tubulin interactions regardless of the species and drug used. The F167Y and F200Y also showed a similar effect that could vary based on the helminth species\u003c/p\u003e"},{"header":"Materials and Methods","content":"\u003cp\u003e\u003cstrong\u003eResearch Questions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe scoping review was done based on the guidelines reported by the PRISMA-ScR (PRISMA Extension for Scoping Reviews) (Tricco et al., 2018) (https://www.prisma-statement.org/scoping). Based on published molecular docking and dynamics studies, this research aims to determine the mechanism of benzimidazole. Specifically, this research answers the following questions:\u003c/p\u003e\n\u003cp\u003e1. What are the \u003cem\u003ein silico \u003c/em\u003eunderpinnings of benzimidazole resistance based on molecular docking and dynamics study?\u003c/p\u003e\n\u003cp\u003e2. What are the consequences of these mutations on the measurement of binding efficiency of the \u0026beta;-tubulins-benzimidazole drug complex?\u003c/p\u003e\n\u003cp\u003e3. What are the consequences of these mutations on the interactions between the \u0026beta;-tubulins and the benzimidazole drug ligand/s?\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSearch Strategy\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eA systematic search was done in three research databases. Scopus (https://www.scopus.com/search), ScienceDirect (https://www.sciencedirect.com/), and MEDLINE via PubMed (https://pubmed.ncbi.nlm.nih.gov/) were searched using the search term \u0026ldquo;Benzimidazole Resistance AND Beta Tubulin AND Molecular Docking.\u0026rdquo; The literature search was done on 17 September 2024. The .ris file of the search results was downloaded.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eStudy Selection, Strategy, and Eligibility\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eUsing the Mendeley citation manager (https://www.mendeley.com), the .ris files were uploaded and utilized for the selection and eligibility assessment. First, duplicates and records with no titles and abstracts (e.g., indexes) were removed. Second, an initial evaluation based on the title and abstract was done. Full-length articles of the studies were accessed for further eligibility appraisal. Figure 1 summarizes the systematic literature search, selection, and eligibility evaluation done.\u003c/p\u003e\n\u003cp\u003eA study was considered eligible for selection if it fulfilled any of the following inclusion criteria:\u003c/p\u003e\n\u003cp\u003e1. Studies that utilized molecular docking in assessing the \u003cem\u003ein silico \u003c/em\u003eeffects of the BZ resistance mutations;\u003c/p\u003e\n\u003cp\u003e2. Studies that utilized molecular docking in assessing the \u003cem\u003ein silico \u003c/em\u003eeffects of the BZ resistance mutations\u003c/p\u003e\n\u003cp\u003eFrom the included studies, papers that did not meet the following criteria were excluded:\u003c/p\u003e\n\u003cp\u003e1. Studies that did not report docking scoring functions (i.e., binding affinities) and/or binding free energies (e.g., MM-PBSA or MM-GBSA);\u003c/p\u003e\n\u003cp\u003e2. Studies that did not report the effects of BZ resistance mutations on the interaction between \u0026beta;-tubulins and benzimidazole drug ligand/s;\u003c/p\u003e\n\u003cp\u003e3. Studies done using non-helminth \u0026beta;-tubulins as macromolecules;\u003c/p\u003e\n\u003cp\u003e4. Studies that did not use commercially available benzimidazole drugs as ligands; and\u003c/p\u003e\n\u003cp\u003e5. Studies that utilized newly designed and synthesized benzimidazole derivatives.\u003c/p\u003e\n\u003cp\u003e6. Research not in the English language.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eRisk of Bias Assessment\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eDue to the \u003cem\u003ein silico \u003c/em\u003eand computational nature of the studies being reviewed, traditional checklists for laboratory experiments are not well-suited as the method of bias assessment. Hence, we developed a simple checklist that is based on the quality of the modeled \u0026beta;-tubulin macromolecule, ligand preparation, docking software, simulation quality, and data analysis utilized. The tool is in the form of a 13-item close-ended questionnaire (Supplementary Table 1). All included studies were evaluated using this tool.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData Acquisition and Synthesis\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe author\u0026rsquo;s name, year of publication, software used in molecular docking and/or system used in molecular dynamics, helminth species of the \u0026beta;-tubulins used as the macromolecule, benzimidazole ligand used, BZ resistance mutation evaluated, docking scoring functions and/or binding free energies of the complex, description of the changes in interactions, and relative resistance-associated effects were the data acquired from the selected studies. Simple descriptive statistics, like counts and frequencies, were used to describe and synthesize the results of this scoping review.\u003c/p\u003e"},{"header":"Results","content":"\u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003eCharacteristics of the studies included\u003c/h2\u003e \u003cp\u003eA total of 37 hits were found in the three databases searched (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Sixteen of these were removed due to duplication. The full text of one article was not accessed. After the eligibility screening, two were removed for not reporting docking scoring function, eight were removed for not using helminth β-tubulins, three were dropped for using newly designed benzimidazole ligands, and one did not report the effects of the docked complexes. In total, six research papers were included in this scoping review. The six research papers included in this review studied several helminth species: \u003cem\u003eHaemonchus conturtos, Trichinella spiralis, Ancylostoma duodenale, Ancylostoma caninum, Ancylostoma ceylanicum, Necator americanus, Trichuris trichiura, Trichuris suis, Anisakis simplex, Ascaris suum, Ascaridia galli, Parascaris equorum, Toxocara canis\u003c/em\u003e, and \u003cem\u003eFasciola hepatica.\u003c/em\u003e The benzimidazole resistance-associated mutations studied included F167Y (TTC, TTT \u0026rarr; TAC, TAT), E198A (GAG, GAA \u0026rarr; GCG, GCA), and F200Y (TTC, TTT \u0026rarr; TAC, TAT).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cb\u003eIn silico\u003c/b\u003e \u003cb\u003eeffects of BZ resistance-associated mutations\u003c/b\u003e\u003c/p\u003e \u003cp\u003eTable\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e summarizes computational studies investigating the molecular mechanisms underlying benzimidazole (BZ) drug resistance in various helminth species. The results highlight the crucial role of specific amino acid residues (e.g., F167, E198, F200) in BZ binding and the potential mechanisms through which mutations in these residues can confer resistance. Mutations in specific amino acid residues (F167, E198, F200) within β-tubulin proteins are frequently associated with BZ drug resistance. These mutations can disrupt the conformation of the β-tubulin active site, destabilize BZ binding, and reduce drug efficacy. Moreover, mutations, particularly at position 198, can lead to the loss of hydrogen bonding interactions between BZ drugs and β-tubulin, contributing to resistance. Further, mutations at positions 167 and 200 may interfere with the opening of the binding site or the internalization of BZ ligands. The impact of mutations on resistance can vary across different helminth species. For example, mutations in F167Y may have a greater impact on strongyle parasites compared to other soil-transmitted helminths. Overall, the results suggest that BZ resistance is primarily attributed to alterations in the β-tubulin binding site, hindering the effective interaction between BZ drugs and their target protein.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eThe key results of the studies included in this scoping review.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"11\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c10\" colnum=\"10\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c11\" colnum=\"11\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSTUDY CODE\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eAUTHORS\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eYEAR PUBLISHED\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eMOLECULAR DOCKING SOFTWARE\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eMOLECULAR DYNAMICS SYSTEM\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eHELMINTH SPECIES\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eβ-TUBULIN EVALUATED\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003eBZ LIGAND USED\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c9\"\u003e \u003cp\u003eDOCKING SCORING FUNCTION(Kcal/mol)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c10\"\u003e \u003cp\u003eBINDING FREE ENERGY (Kcal/mol)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c11\"\u003e \u003cp\u003eRESISTANCE EFFECTS NOTED\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"27\" rowspan=\"28\"\u003e \u003cp\u003eS1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\" morerows=\"27\" rowspan=\"28\"\u003e \u003cp\u003eAguayo-Ortiz et al. (\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2013\u003c/span\u003eb)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\" morerows=\"27\" rowspan=\"28\"\u003e \u003cp\u003e2013\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\" morerows=\"27\" rowspan=\"28\"\u003e \u003cp\u003eAutoDock 4.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\" morerows=\"27\" rowspan=\"28\"\u003e \u003cp\u003eGROMACS 4.5.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\" morerows=\"27\" rowspan=\"28\"\u003e \u003cp\u003e\u003cem\u003eHaemonchus conturtos\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\" morerows=\"6\" rowspan=\"7\"\u003e \u003cp\u003eWild-type\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eAlbendazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-8.10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e-68.41\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\" morerows=\"27\" rowspan=\"28\"\u003e \u003cp\u003eThe mutated and unsusceptible β-tubulin models suggest that the primary cause of BZ resistance is likely due to an amino acid modification at position 198, resulting in the loss of hydrogen bonding interactions. Conversely, the substitution of phenylalanine for tyrosine at positions 167 and 200 implies that the inhibitory mechanism may occur either during the opening of the binding site or the internalization of the ligand.\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eCarbendazim\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-6.98\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e-55.08\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eOxibendazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-8.17\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e-67.27\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eParbendazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-8.09\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e-65.59\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eLuxabendazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-9.46\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e-64.77\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eMebendazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-9.43\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e-60.53\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eNocodazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-9.36\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e-64.61\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c7\" morerows=\"6\" rowspan=\"7\"\u003e \u003cp\u003eF167Y\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eAlbendazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-8.07\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\" morerows=\"20\" rowspan=\"21\"\u003e \u003cp\u003eN/A\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eCarbendazim\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-7.02\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eOxibendazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-8.09\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eParbendazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-8.15\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eLuxabendazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-9.26\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eMebendazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-9.46\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eNocodazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-9.35\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c7\" morerows=\"6\" rowspan=\"7\"\u003e \u003cp\u003eE198A\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eAlbendazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-7.03\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eCarbendazim\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-6.37\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eOxibendazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-6.66\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eParbendazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-7.05\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eLuxabendazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-8.82\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eMebendazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-8.26\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eNocodazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-7.99\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c7\" morerows=\"6\" rowspan=\"7\"\u003e \u003cp\u003eF200Y\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eAlbendazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-8.22\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eCarbendazim\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-7.19\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eOxibendazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-8.28\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eParbendazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-8.26\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eLuxabendazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-9.62\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eMebendazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-9.48\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eNocodazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-9.35\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"6\" rowspan=\"7\"\u003e \u003cp\u003eS2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\" morerows=\"6\" rowspan=\"7\"\u003e \u003cp\u003eAguayo-Ortiz et al. (\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2013\u003c/span\u003ea)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\" morerows=\"6\" rowspan=\"7\"\u003e \u003cp\u003e2013\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\" morerows=\"6\" rowspan=\"7\"\u003e \u003cp\u003eAutoDock 4.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\" morerows=\"6\" rowspan=\"7\"\u003e \u003cp\u003eGROMACS 4.5.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\" morerows=\"6\" rowspan=\"7\"\u003e \u003cp\u003e\u003cem\u003eTrichinella spiralis\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\" morerows=\"6\" rowspan=\"7\"\u003e \u003cp\u003eWild-type\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eAlbendazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-7.70\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e-53.67\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\" morerows=\"6\" rowspan=\"7\"\u003e \u003cp\u003eThe modeling study found that the binding site for BZ aligns with previous experimental findings. This site includes amino acids linked to resistance mutations (F167Y, E198A, and F200Y), and overlaps with the colchicine-binding site. Molecular docking and dynamics calculations of BZ drugs reveal that they are stabilized within the binding site primarily through hydrogen bonds with specific amino acids (e.g., Thr165, Glu198, Cys239 and Gln134).\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eCarbendazim\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-7.05\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e-21.74\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eOxibendazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-7.41\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e-31.92\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eParbendazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-7.93\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e-37.20\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eLuxabendazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-9.41\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e-44.71\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eMebendazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-8.73\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e-43.04\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eNocodazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-8.82\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e-34.34\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"3\" rowspan=\"4\"\u003e \u003cp\u003eS3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\" morerows=\"3\" rowspan=\"4\"\u003e \u003cp\u003eJones et al. (\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2022\u003c/span\u003ea)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\" morerows=\"3\" rowspan=\"4\"\u003e \u003cp\u003e2022\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\" morerows=\"3\" rowspan=\"4\"\u003e \u003cp\u003eAutodock vina v1.1.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\" morerows=\"3\" rowspan=\"4\"\u003e \u003cp\u003eMolecular Operating Environment (MOE) 2020.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\" morerows=\"3\" rowspan=\"4\"\u003e \u003cp\u003e\u003cem\u003eAscaris suum\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eWild-type\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eAlbendazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-8.46\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\" morerows=\"3\" rowspan=\"4\"\u003e \u003cp\u003eN/A\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\" morerows=\"3\" rowspan=\"4\"\u003e \u003cp\u003eIn silico docking studies suggest that BZ drugs can bind to all Ascarid β-tubulin isotypes. Ascarid β-tubulin isotype A was further analyzed using molecular dynamics simulations. These simulations revealed the critical role of amino acid E198 in BZ-β-tubulin interactions. Mutations at E198A and F200Y were found to alter benzimidazole binding, while the F167Y mutation had no significant impact.\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eF167Y\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eAlbendazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-8.04\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eE198A\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eAlbendazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-8.53\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eF200Y\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eAlbendazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-8.16\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"15\" rowspan=\"16\"\u003e \u003cp\u003eS4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\" morerows=\"15\" rowspan=\"16\"\u003e \u003cp\u003eJones et al. (\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2022\u003c/span\u003eb)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\" morerows=\"15\" rowspan=\"16\"\u003e \u003cp\u003e2022\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\" morerows=\"15\" rowspan=\"16\"\u003e \u003cp\u003eAutodock vina v1.1.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\" morerows=\"15\" rowspan=\"16\"\u003e \u003cp\u003eMolecular Operating Environment (MOE) version 2020.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\" morerows=\"3\" rowspan=\"4\"\u003e \u003cp\u003e\u003cem\u003eAncylostoma duodenale\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eWild-type\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eAlbendazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-8.55\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\" morerows=\"15\" rowspan=\"16\"\u003e \u003cp\u003eN/A\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\" morerows=\"15\" rowspan=\"16\"\u003e \u003cp\u003eThe study indicates that BZ acts through similar mechanisms in various helminth species. The amino acid E198 plays a crucial role in BZ binding. However, the Q134\u0026ndash;F167Y interaction observed in \u003cem\u003eAncylostoma caninum\u003c/em\u003e, along with the presence of the F167Y SNP in susceptible \u003cem\u003eAscaris\u003c/em\u003e populations and its reduced frequency after BZ treatment in \u003cem\u003eTrichuris trichiura\u003c/em\u003e from previous studies, suggests that mutations in F167Y may have a greater impact on strongyle parasites compared to other STHs.\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eF167Y\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eAlbendazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-8.19\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eE198A\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eAlbendazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-8.14\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eF200Y\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eAlbendazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-7.72\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c6\" morerows=\"3\" rowspan=\"4\"\u003e \u003cp\u003e\u003cem\u003eTrichuris trichiura\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eWild-\u003c/p\u003e \u003cp\u003eType\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eAlbendazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-8.53\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eF167Y\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eAlbendazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-9.82\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eE198A\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eAlbendazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-7.78\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eF200Y\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eAlbendazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-10.48\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u003cem\u003eAnisakis simplex\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eWild-type\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eAlbendazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-7.74\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u003cem\u003eAscaridia galli\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eWild-type\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eAlbendazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-8.25\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u003cem\u003eParascaris equorum\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eWild-type\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eAlbendazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-8.19\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u003cem\u003eToxocara canis\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eWild-type\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eAlbendazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-8.82\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u003cem\u003eAncylostoma caninum\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eWild-type\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eAlbendazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-7.94\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u003cem\u003eAncylostoma ceylanicum\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eWild-type\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eAlbendazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-8.29\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u003cem\u003eNecator americanus\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eWild-type\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eAlbendazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-7.54\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u003cem\u003eTrichuris suis\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eWild-type\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eAlbendazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-8.53\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"5\" rowspan=\"6\"\u003e \u003cp\u003eS5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\" morerows=\"5\" rowspan=\"6\"\u003e \u003cp\u003eOlivares-Ferretti et al. (2023)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\" morerows=\"5\" rowspan=\"6\"\u003e \u003cp\u003e2023\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\" morerows=\"5\" rowspan=\"6\"\u003e \u003cp\u003eAutoDock Vina program in PyRx software\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\" morerows=\"5\" rowspan=\"6\"\u003e \u003cp\u003eN/A\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\" morerows=\"5\" rowspan=\"6\"\u003e \u003cp\u003e\u003cem\u003eFasciola hepatica\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eWild-type isotype 1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eTriclabendazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-6.87\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\" morerows=\"5\" rowspan=\"6\"\u003e \u003cp\u003eN/A\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\" morerows=\"5\" rowspan=\"6\"\u003e \u003cp\u003eThe nucleotide binding site on \u003cem\u003eF. hepatica\u003c/em\u003e β-tubulin exhibits a higher affinity for ligands than other known binding sites, such as colchicine, albendazole, the T7 loop, and pβVII. Ligand binding to the polymerization site of β-tubulin can disrupt microtubule formation. Triclabendazole demonstrated significantly higher binding affinity than other ligands across all β-tubulin isotypes. Computational analysis has provided new insights into the mechanism of action of triclabendazole on \u003cem\u003eF. hepatica\u003c/em\u003e β-tubulin.\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eWild-type isotype 2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eTriclabendazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-6.40\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eWild-type isotype 3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eTriclabendazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-6.38\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eWild-type isotype 4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eTriclabendazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-6.33\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eWild-type isotype 5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eTriclabendazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-6.17\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eWild-type isotype 6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eTriclabendazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-6.48\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eS6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eYashica et al. (2024)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e2024\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eAutoDock Tools in MGL tools 1.5.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eN/A\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u003cem\u003eHaemonchus contortus\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eWild-type\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eAlbendazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e-8.51\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eN/A\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003eThe \u003cem\u003ein silico\u003c/em\u003e study suggests that mutations at amino acid position 200 can disrupt the conformation of the \u003cem\u003eH. conturtos\u003c/em\u003e β-tubulin active site and destabilize albendazole binding.\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eEffects of Mutations on Docking Scoring Function and Binding Free Energy\u003c/h3\u003e\n\u003cp\u003eBased on the results of the included studies, the BZ resistance mutations often lead to a decrease in the docking scoring function and binding free energy, indicating a weaker interaction between the BZ drug and the mutated β-tubulin protein. However, the magnitude of the effect can vary depending on the specific mutation and the BZ drug involved. While F167Y mutations can sometimes decrease binding affinity, their effect is often less pronounced compared to mutations at other residues. Also, the impact of F167Y may vary across different helminth species. Meanwhile, the E198A mutation consistently leads to a marked decrease in binding affinity, suggesting a crucial role of this residue in BZ binding. The E198A mutation likely disrupts hydrogen bonding interactions between the BZ drug and β-tubulin. The F200Y mutations generally result in a moderate decrease in binding affinity. This mutation may alter the conformation of the β-tubulin active site, affecting drug binding. The combined effects of multiple mutations can further reduce binding affinity and enhance resistance. Likewise, the impact of mutations may vary depending on the specific BZ drug being considered. Overall, the results suggest that mutations in these key residues can disrupt the structural and electrostatic interactions between BZ drugs and β-tubulin, leading to reduced binding affinity and ultimately, drug resistance.\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eThis scoping review was conducted with the aim of unraveling the mechanism of BZ resistance based on published research on molecular docking and dynamics. MEDLINE via PubMed, Scopus, and Science Direct were searched. A total of six eligible studies were included, which encompassed research that utilized β-tubulins from several helminth species and numerous benzimidazole ligands. Of the BZ mutations studied, E198A showed that it can drive down the binding affinity of BZ ligand-β-tubulin interactions. The F167Y and F200Y also showed a similar effect that could vary based on the helminth species. The F200Y mutation can alter the conformation of the β-tubulin active site, negatively affecting drug binding.\u003c/p\u003e \u003cp\u003eThe studies included in this scoping review assessed both wild-type and mutated helminth β-tubulins. The three canonical BZ resistance mutations\u0026mdash;F167Y, E198A, and F200Y\u0026mdash;that are induced by SNPs were all evaluated (Furtado et al., \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). These mutations have been reported in many helminth species of public and veterinary health concern from many parts of the world (Diawara et al., \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2013\u003c/span\u003e; George et al., \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Jimenez Castro et al., \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). Hence, their inclusion in computational studies was warranted. However, their actual contribution to conferring resistance has been put into question (Lacey and Gill, \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e1994\u003c/span\u003e). Also, several recent evidence of alternative resistance mechanisms, such as enzymatic biotransformation of drugs using UDP-glycosyltransferases and long non-coding RNA interference (Chen et al., \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; Dimunov\u0026aacute; et al., \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2022\u003c/span\u003e), have been reported. However, laboratory research using gene editing that encoded resistance-associated mutations in \u003cem\u003eCaenorhabditis elegans\u003c/em\u003e showed their actual potential to confer resistance, as previously mentioned (Dilks et al., \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2021\u003c/span\u003e, \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Particularly, varying mutations that have been reported in amino acid position 198 have been reported to make edited \u003cem\u003eC. elegans\u003c/em\u003e significantly more resistant to benzimidazole treatment compared to their wild-type counterpart (Dilks et al., \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). This result echoes the finding in this systematic review that the E198A mutation had a consistent negative effect on the binding of BZ drugs and helminth β-tubulins indicating their important role in conferring resistance.\u003c/p\u003e \u003cp\u003eThis scoping review has several limitations. First, only a few accessible databases were searched, hence there might be other research not indexed in these databases that were missed. However, searching only known indexing databases, like MEDLINE via PubMed and Scopus, ensures that only quality research papers from reputable journals are included. Second, the \u003cem\u003ein silico\u003c/em\u003e nature of the research targeted for this review may lead to conclusions of insufficient evidence for the conclusions that they present due to the lack of laboratory confirmation. However, advances in computational biology have assured that the predictions are of high quality and accuracy, particularly for molecular docking and dynamics studies (Santos et al., \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Singh et al., \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2022\u003c/span\u003e).\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eThe computational studies included in this scoping review provide valuable insights into the molecular mechanisms underlying BZ drug resistance in various helminth species. The results consistently demonstrate that mutations in specific amino acid residues within β-tubulin proteins, particularly F167, E198, and F200, play a critical role in conferring resistance. These mutations disrupt the structural and electrostatic interactions between BZ drugs and helminth β-tubulin, leading to reduced binding affinity and ultimately, drug resistance. While the impact of these mutations can vary depending on the specific helminth species and the BZ drug involved, the overall findings highlight the importance of targeting these residues for the development of novel anthelmintic strategies to address emerging drug resistance. Future studies could explore additional mutations, investigate the interactions between BZ drugs and other β-tubulin isoforms, or investigate the potential for combination therapies to overcome resistance.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eSupplementary Materials\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eSupplementary tables and figures are available at [DOI}.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgments\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNone.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNo funding was received to assist with the preparation of this manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflict of Interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe author declares that there is no conflict of any financial or non-financial interests regarding the publication of this paper.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthics Approval\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent to participate\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData Availability\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eAguayo-Ortiz, R., M\u0026eacute;ndez-Lucio, O., Medina-Franco, J.L., Castillo, R., Y\u0026eacute;pez-Mulia, L., Hern\u0026aacute;ndez-Luis, F., Hern\u0026aacute;ndez-Campos, A., 2013. Towards the identification of the binding site of benzimidazoles to \u0026beta;-tubulin of Trichinella spiralis: Insights from computational and experimental data. Journal of Molecular Graphics and Modelling 41, 12\u0026ndash;19. https://doi.org/10.1016/j.jmgm.2013.01.007\u003c/li\u003e\n\u003cli\u003eChen, X., Wang, T., Guo, W., Yan, X., Kou, H., Yu, Y., Liu, C., Gao, W., Wang, W., Wang, R., 2024. Transcriptome reveals the roles and potential mechanisms of lncRNAs in the regulation of albendazole resistance in Haemonchus contortus. BMC Genomics 25, 188. https://doi.org/10.1186/s12864-024-10096-6\u003c/li\u003e\n\u003cli\u003eDiawara, A., Halpenny, C.M., Churcher, T.S., Mwandawiro, C., Kihara, J., Kaplan, R.M., Streit, T.G., Idaghdour, Y., Scott, M.E., Bas\u0026aacute;\u0026ntilde;ez, M.-G., Prichard, R.K., 2013. Association between Response to Albendazole Treatment and \u0026beta;-Tubulin Genotype Frequencies in Soil-transmitted Helminths. PLoS Negl Trop Dis 7, e2247. https://doi.org/10.1371/journal.pntd.0002247\u003c/li\u003e\n\u003cli\u003eDilks, C.M., Hahnel, S.R., Sheng, Q., Long, L., McGrath, P.T., Andersen, E.C., 2020. Quantitative benzimidazole resistance and fitness effects of parasitic nematode beta-tubulin alleles. Int J Parasitol Drugs Drug Resist 14, 28\u0026ndash;36. https://doi.org/10.1016/j.ijpddr.2020.08.003\u003c/li\u003e\n\u003cli\u003eDilks, C.M., Koury, E.J., Buchanan, C.M., Andersen, E.C., 2021. Newly identified parasitic nematode beta-tubulin alleles confer resistance to benzimidazoles. International Journal for Parasitology: Drugs and Drug Resistance 17, 168\u0026ndash;175. https://doi.org/10.1016/j.ijpddr.2021.09.006\u003c/li\u003e\n\u003cli\u003eDimunov\u0026aacute;, D., Navr\u0026aacute;tilov\u0026aacute;, M., Kellerov\u0026aacute;, P., Ambrož, M., Sk\u0026aacute;lov\u0026aacute;, L., Matou\u0026scaron;kov\u0026aacute;, P., 2022. The induction and inhibition of UDP-glycosyltransferases in Haemonchus contortus and their role in the metabolism of albendazole. International Journal for Parasitology: Drugs and Drug Resistance 19, 56\u0026ndash;64. https://doi.org/10.1016/j.ijpddr.2022.06.001\u003c/li\u003e\n\u003cli\u003eFurtado, L.F.V., De Paiva Bello, A.C.P., Rabelo, \u0026Eacute;.M.L., 2016. Benzimidazole resistance in helminths: From problem to diagnosis. Acta Tropica 162, 95\u0026ndash;102. https://doi.org/10.1016/j.actatropica.2016.06.021\u003c/li\u003e\n\u003cli\u003eGeorge, S., Suwondo, P., Akorli, J., Otchere, J., Harrison, L.M., Bilguvar, K., Knight, J.R., Humphries, D., Wilson, M.D., Caccone, A., Cappello, M., 2022. Application of multiplex amplicon deep-sequencing (MAD-seq) to screen for putative drug resistance markers in the Necator americanus isotype-1 \u0026beta;-tubulin gene. Sci Rep 12, 11459. https://doi.org/10.1038/s41598-022-15718-1\u003c/li\u003e\n\u003cli\u003eJimenez Castro, P.D., Venkatesan, A., Redman, E., Chen, R., Malatesta, A., Huff, H., Zuluaga Salazar, D.A., Avramenko, R., Gilleard, J.S., Kaplan, R.M., 2021. Multiple drug resistance in hookworms infecting greyhound dogs in the USA. International Journal for Parasitology: Drugs and Drug Resistance 17, 107\u0026ndash;117. https://doi.org/10.1016/j.ijpddr.2021.08.005\u003c/li\u003e\n\u003cli\u003eJones, B.P., van Vliet, A.H.M., LaCourse, E.J., Betson, M., 2022. Identification of key interactions of benzimidazole resistance-associated amino acid mutations in Ascaris \u0026beta;-tubulins by molecular docking simulations. Sci Rep 12, 13725. https://doi.org/10.1038/s41598-022-16765-4\u003c/li\u003e\n\u003cli\u003eLacey, E., Gill, J.H., 1994. Biochemistry of benzimidazole resistance. Acta Trop 56, 245\u0026ndash;262. https://doi.org/10.1016/0001-706x(94)90066-3\u003c/li\u003e\n\u003cli\u003eNg\u0026rsquo;etich, A.I., Amoah, I.D., Bux, F., Kumari, S., 2023. Anthelmintic resistance in soil-transmitted helminths: One-Health considerations. Parasitol Res 123, 62. https://doi.org/10.1007/s00436-023-08088-8\u003c/li\u003e\n\u003cli\u003eSantos, L.H.S., Ferreira, R.S., Caffarena, E.R., 2019. Integrating Molecular Docking and Molecular Dynamics Simulations, in: De Azevedo, W.F. (Ed.), Docking Screens for Drug Discovery, Methods in Molecular Biology. Springer New York, New York, NY, pp. 13\u0026ndash;34. https://doi.org/10.1007/978-1-4939-9752-7_2\u003c/li\u003e\n\u003cli\u003eSchwenkenbecher, J.M., Albonico, M., Bickle, Q., Kaplan, R.M., 2007. Characterization of beta-tubulin genes in hookworms and investigation of resistance-associated mutations using real-time PCR. Mol Biochem Parasitol 156, 167\u0026ndash;174. https://doi.org/10.1016/j.molbiopara.2007.07.019\u003c/li\u003e\n\u003cli\u003eSingh, S., Bani Baker, Q., Singh, D.B., 2022. Molecular docking and molecular dynamics simulation, in: Bioinformatics. Elsevier, pp. 291\u0026ndash;304. https://doi.org/10.1016/B978-0-323-89775-4.00014-6\u003c/li\u003e\n\u003cli\u003eTenorio, J.C.B., 2023. Canine hookworms in the Philippines\u0026mdash;Very common but very much neglected in veterinary research. Frontiers in Veterinary Science 10.\u003c/li\u003e\n\u003cli\u003eTenorio, J.C.B., Tabios, I.K.B., Inpankaew, T., Yba\u0026ntilde;ez, A.P., Tiwananthagorn, S., Tangkawattana, S., Suttiprapa, S., 2024. Ancylostoma ceylanicum and other zoonotic canine hookworms: neglected public and animal health risks in the Asia\u0026ndash;Pacific region. Animal Diseases 4, 11. https://doi.org/10.1186/s44149-024-00117-y\u003c/li\u003e\n\u003cli\u003eTricco, A.C., Lillie, E., Zarin, W., O\u0026rsquo;Brien, K.K., Colquhoun, H., Levac, D., Moher, D., Peters, M.D.J., Horsley, T., Weeks, L., Hempel, S., Akl, E.A., Chang, C., McGowan, J., Stewart, L., Hartling, L., Aldcroft, A., Wilson, M.G., Garritty, C., Lewin, S., Godfrey, C.M., Macdonald, M.T., Langlois, E.V., Soares-Weiser, K., Moriarty, J., Clifford, T., Tun\u0026ccedil;alp, \u0026Ouml;., Straus, S.E., 2018. PRISMA Extension for Scoping Reviews (PRISMA-ScR): Checklist and Explanation. Ann Intern Med 169, 467\u0026ndash;473. https://doi.org/10.7326/M18-0850\u003c/li\u003e\n\u003cli\u003eTroCCAP, 2019. Guidelines for the diagnosis, treatment and control of canine endoparasites in the tropics, 2nd ed.\u003c/li\u003e\n\u003cli\u003eVenkatesan, A., Castro, P.D.J., Morosetti, A., Horvath, H., Chen, R., Redman, E., Dunn, K., Collins, J.B., Fraser, J.S., Andersen, E.C., Kaplan, R.M., Gilleard, J.S., 2023. Molecular evidence of widespread benzimidazole drug resistance in Ancylostoma caninum from domestic dogs throughout the USA and discovery of a novel \u0026beta;-tubulin benzimidazole resistance mutation. PLOS Pathogens 19, e1011146. https://doi.org/10.1371/journal.ppat.1011146\u003c/li\u003e\n\u003cli\u003eVon Samson-Himmelstjerna, G., Blackhall, W.J., McCARTHY, J.S., Skuce, P.J., 2007. Single nucleotide polymorphism (SNP) markers for benzimidazole resistance in veterinary nematodes. Parasitology 134, 1077\u0026ndash;1086. https://doi.org/10.1017/S0031182007000054\u003c/li\u003e\n\u003cli\u003eWhittaker, J.H., Carlson, S.A., Jones, D.E., Brewer, M.T., 2017. Molecular mechanisms for anthelmintic resistance in strongyle nematode parasites of veterinary importance. J. vet. Pharmacol. Therap. 40, 105\u0026ndash;115. https://doi.org/10.1111/jvp.12330\u003c/li\u003e\n\u003cli\u003eWorld Health Organization, 2011. Helminth control in school-age children : a guide for managers of control programmes. World Health Organization.\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"highlight":"","institution":"University of Southern Mindanao","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Drug resistance, computational biology, docking, helminths ","lastPublishedDoi":"10.21203/rs.3.rs-5476123/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-5476123/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eBackground: \u003c/strong\u003eBenzimidazole (BZ) resistance remains an emerging grave concern in helminths of public and veterinary health concerns. Resistance against BZ drugs is due to mutations that change the amino acid comprising the β-tubulin protein, which negatively affects its interactions with BZ drug molecules. Several in silico modeling studies have been published to decipher the precise mechanism of BZ resistance, but inconsistencies on the resistance consequence mutations confer and the effect of different BZ ligands have led to further confusion regarding the exact mechanism of resistance. Hence, this scoping review was done to unravel the mechanism of BZ resistance based on published research on molecular docking and dynamics.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMethods: \u003c/strong\u003eA scoping review was conducted in ScienceDirect, MEDLINE via PubMed and Scopus using the search term “Benzimidazole Resistance AND Beta Tubulin AND Molecular Docking”. A total of 37 hits were recovered and from these 6 were included after selection, inclusion, and risk of bias assessment.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults:\u003c/strong\u003e The six research papers included in this review studied several helminth species: \u003cem\u003eHaemonchus conturtos, Trichinella spiralis, Ancylostoma duodenale, Ancylostoma caninum, Ancylostoma ceylanicum, Necator americanus, Trichuris trichiura, Trichuris suis, Anisakis simplex, Ascaris suum, Ascaridia galli, Parascaris equorum, Toxocara canis\u003c/em\u003e, and \u003cem\u003eFasciola hepatica\u003c/em\u003e. The benzimidazole resistance-associated mutations studied included F167Y (TTC, TTT → TAC, TAT), E198A (GAG, GAA → GCG, GCA), and F200Y (TTC, TTT → TAC, TAT). The results show that the E198A can markedly reduce the binding affinity of BZ ligand-β-tubulin interactions. The F167Y and F200Y also showed a similar effect that could vary based on the helminth species. The F200Y mutation can alter the conformation of the β-tubulin active site, negatively affecting drug binding.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusion:\u003c/strong\u003e While the impact of these mutations can vary depending on the specific helminth species and the BZ drug involved, the overall findings highlight the importance of targeting these residues for the development of novel anthelmintic strategies to address emerging drug resistance.\u003c/p\u003e","manuscriptTitle":"Molecular docking and dynamics as a tool to study benzimidazole resistance in helminths: A scoping review","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-11-19 02:18:33","doi":"10.21203/rs.3.rs-5476123/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"91f6e695-37f5-4422-8e4c-339e609f2213","owner":[],"postedDate":"November 19th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":40408140,"name":"Parasitology"}],"tags":[],"updatedAt":"2024-11-19T02:18:33+00:00","versionOfRecord":[],"versionCreatedAt":"2024-11-19 02:18:33","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-5476123","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-5476123","identity":"rs-5476123","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}