Review Protocol: Benzotriazole in cancer: preclinical evidence and structure–activity relationship insights (systematic review)
Abstract
This review protocol outlines the methodology for investigating the preclinical evidence and structure-activity relationship of benzotriazole derivatives in cancer. Benzotriazole derivatives are promising anticancer agents in preclinical studies, but the existing evidence is scattered across various experimental models, necessitating a systematic synthesis. The review's objectives include identifying in vitro and in vivo preclinical studies, classifying the derivatives chemically, summarizing their anticancer mechanisms, and assessing the risk of bias using the QUIN tool for in vitro studies and the SYRCLE tool for in vivo studies.
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Review Protocol: Benzotriazole in cancer: preclinical evidence and structure–activity relationship insights Authors Gabriel Mardale1,2, Alexandra Prodea1,2, Andreea Munteanu1,2, Mihaela Jorgovan1,2, Sabina Mardale3, Victor Cristian Dumitrascu3, Codruta Soica1,2 Affiliations (1) Faculty of Pharmacy, “Victor Babes” University of Medicine and Pharmacy, Eftimie Murgu Square, No. 2, 300041 Timisoara, Romania (2) Research Center for Experimental Pharmacology and Drug Design (X-Pharm Design), “Victor Babes” University of Medicine and Pharmacy, Eftimie Murgu Square, No. 2, 300041 Timișoara, Romania (3) Timis County Emergency Clinical Hospital Pius Brinzeu, Liviu Rebreanu 156, Timișoara 300723, Romania Background Benzotriazole derivatives have shown promising anticancer activity in preclinical studies (in vitro and in vivo); however, evidence is scattered across heterogeneous experimental models. A systematic synthesis is needed to evaluate structural classes, mechanisms of action, and methodological quality. Objectives 1. Identify in vitro and in vivo preclinical studies on benzotriazole derivatives in cancer 2. Classify derivatives based on chemical structure 3. Summarize anticancer mechanisms 4. Assess risk of bias in included studies (in vitro and in vivo) Eligibility Criteria Models: in vitro and in vivo cancer models Intervention: benzotriazole and benzotriazole derivatives Comparators: untreated controls, positive controls such as reference anticancer drugs Outcomes: cytotoxicity, tumor growth, toxicity, mechanisms Study design: Original studies Language: English only Timeframe: January 2010 – May 2025 Exclusion Criteria duplicates, secondary studies, studies lacking anticancer evaluation, studies based only on in silico analysis and articles that could not be retrieved in full. Databases PubMed/MEDLINE, Scopus, and Web of Science.
Search Strategy Search terms benzotriazole, benzotriazole derivatives, 1H-benzotriazole, in vivo, in vitro, animal model, pharmacological effect, therapeutic effect, anticancer. Search sequences PubMed: (benzotriazole OR benzotriazole derivatives OR 1H-benzotriazole) AND (in vivo OR "in vitro" OR "animal model" OR "pharmacological effect" OR "therapeutic effect" OR anticancer) Filters: Search field (title/abstract); Article language (English) WOS: (benzotriazole OR benzotriazole derivatives OR 1H-benzotriazole) AND (in vivo OR “in vitro” OR "animal model" OR "pharmacological effect" OR "therapeutic effect" OR “anticancer”) Filters: Search field (topic); Language (English) Scopus: (benzotriazole OR benzotriazole derivatives OR 1H-benzotriazole) AND (in vivo OR “in vitro” OR "animal model" OR "pharmacological effect" OR "therapeutic effect" OR anticancer) Filters: Search within (article title, abstract and keywords); Language (English) Databases searched PubMed/Medline WOS Scopus Other sources - Part of journals searched Title, abstract Years of search January 2010May 2025 Language English language only Types of studies to be included Original articles Study Selection Two reviewers independently screened titles/abstracts (G.M. and A.M.) and full texts (G.M. and A.P.). Disagreements were resolved through discussion. Data Extraction Data extracted included publication details, compound identifiers, experimental models, dosing, outcomes, mechanisms and toxicity. Extraction was performed by one reviewer and verified by a second (G.M. and A.M.). Risk of Bias Assessment
In vitro studies were assessed using the QUIN tool by G.M. and A.P. In vivo studies were assessed using the SYRCLE risk-of-bias tool by G.M. and A.M. Data Synthesis Due to methodological heterogeneity, results were synthesized narratively and summarized in structured tables. Protocol Registration Prospective registration in PROSPERO was not possible because the review had already commenced. The protocol review was retrospectively registered and is available at: 10.5281/zenodo.17950716 Deviations from Protocol No deviations from this protocol occurred during the conduct of the review. Ethics No ethical approval was required. Dissemination Results are intended for publication in a peer-reviewed journal. Contributions G.M. and A.P. conceived the review; G.M. and A.P. performed the search; G.M. and A.M. performed the initial screening; G.M. and A.P. performed the secondary screening. G.M. and A.M. extracted the data; G.M. and A.P. conducted the risk of bias for in vitro studies (QUIN tool); G.M. and A.M. conducted the risk of bias for animal studies (SYRCLE tool). All authors contributed to the conceptualization, drafting, and critical revision of the protocol and approved the final version