Regulatory Applications Research Articles

Increasing chemical coverage, accuracy, and reproducibility of the processing method for polar organic chemical integrative samplers.

Despite their diffusion in research studies, passive samplers are rarely used in regulatory applications. To expand the employment of passive samplers in regulatory environmental studies, standardized procedures for processing each sampler type should be proposed and accepted, but currently, each study develops its own protocol based on previous knowledge and specific needs. In this work, six identical polar organic chemical integrative samplers in seawater were deployed to understand the importance of the sorbent transfer method prior to the elution step. A common "wet transfer" with ultra-pure water was compared to a less diffused "dry-transfer," assessing recoveries and matrix effects of 38 target compounds of emerging concern, including polar pesticides, recreational and food-related substances, pharmaceuticals, industrial additives, and ultra-violet (UV) filters. The dry-transfer procedure generally allowed better recoveries, especially for the more polar compounds, without affecting matrix effects (which remained in the range 40-130%). Along with the recovery assessment, the analysis of the non-spiked sorbent extracts revealed traces of many of the targeted emerging contaminants, quantifying perfluorooctanoic acid, UV-filters, carbamazepine, diclofenac, and triclosan. Furthermore, other compounds were found below their limits of quantitation. Ten analytes were detected only in the extracts of the dry-transferred passive samplers, highlighting the importance of applying this protocol, especially when dealing with polar compounds. This refined processing method, therefore, permits a more standardized and reproducible strategy, at the same time enlarging the set of analytes which could be detected and quantified.

Erratum for Huang et al., "Insights into the regulatory mechanisms and application prospects of the transcription factor Cra".

Erratum for Huang et al., "Insights into the regulatory mechanisms and application prospects of the transcription factor Cra".

Leveraging Model Master Files for Long-Acting Injectables.

The U.S. Food and Drug Administration (FDA) and the Center for Research on Complex Generics (CRCG) hosted a public workshop on May 2-3, 2024, titled "Considerations and Potential Regulatory Applications for a Model Master File (Lachaine et al Can J Psychiatry. 60:S40-S47, 2015)". The workshop aimed to discuss the application of the Model Master File (MMF) concept in regulatory submissions that contain model integrated evidence (MIE), improving model sharing, model standardization, regulatory consistency, and regulatory efficiency. On Day 1, there was a session dedicated to MMF applications for long-acting injectables (LAIs). This perspective summarizes presentations, panel discussion, and small group discussion for the potential applications of MMFs in LAI product development, including case studies and potential situations in which MMFs can support regulatory submissions. The scientific presentations discussed the application of MMFs in mechanistic physiologically based pharmacokinetic (PBPK), multiphysics simulation, and population pharmacokinetics (popPK) models, as well as the potential utility of a model-integrated bioequivalence (MI-BE) framework. Additionally, challenges and considerations of implementing MMFs for LAIs were discussed in the panel and small groups. The anticipated benefits of MMFs are recognized among model developers, industries, and regulators.

A 2024 Update on US FDA Implementation of Partial Area Under the Curve Into Bioavailability and Bioequivalence Assessments.

Comparisons of maximum drug concentration (Cmax) and total area under the concentration vs. time curve (AUC) may be inadequate for bioavailability (BA)/bioequivalence (BE) assessments in cases where the shape of the pharmacokinetic (PK) profile of a drug impacts the clinical performance. In such cases, partial area under the concentration vs. time curve (pAUC) has been recommended by regulatory agencies to support BA or BE assessments as a measure of drug exposure over specified time intervals of clinical relevance. This white paper serves as an update to the previously published white paper by Fang et al. at the US Food and Drug Administration (FDA), which introduced the general framework to provide pAUC recommendations. Since August 2020, 18 product-specific guidances (PSGs) have been published or revised using the general framework to provide consistent, science- and risk-based pAUC recommendations. Notable regulatory examples of pAUC applications discussed include loxapine inhalation powder, leuprolide long-acting injectables (LAIs), and goserelin LAIs. This paper discusses recent applications of pAUC in the United States, highlights key examples of pAUC recommendations for regulatory applications, and provides insights about areas for global harmonization of pAUC recommendations.

Regulatory mechanisms and applications of Lactobacillus biofilms in the food industry.

Lactobacillus is widely recognized for its probiotic benefits and has been widely used in food production. While biofilms are typically associated with pathogenic bacteria, they also served as a self-protective mechanism formed by microorganisms in an adverse environments. In recent years, relevant studies have revealed the excellent characteristics of Lactobacillus biofilms, offering new insights into their potential applications in the food industry. The Lactobacillus biofilms is important in improving fermentation processes and enhancing the resilience of Lactobacillus in various conditions. This paper reviews how quorum sensing regulates the formation of Lactobacillus biofilms and explores their roles in stress resistance, bacteriostasis and food production. Additionally, it highlights the emerging concept of fourth-generation probiotics, developed through biofilm technology, as a novel approach to probiotic applications.

The Use of MPS in Three Rs and Regulatory Applications: Perspectives From Developers on Stakeholder Responsibilities.

Increasing the use of microphysiological systems (MPS) in Three Rs and regulatory applications is a nuanced but important goal, which would also help increase their scientific impact. There are three distinct and important stakeholder groups that each play a unique role in expediting the use of MPS for regulatory purpose - namely, commercial MPS developers, end-users and regulators. Additionally, non-profit organisations, such as the 3Rs Collaborative (3RsC), can help coordinate these efforts. This paper introduces the MPS Initiative, as organised by the 3RsC, and clarifies the potential for MPS to benefit all Three Rs. Key differences in the use of MPS-derived data for regulatory evidence of efficacy versus safety, and for various other contexts of use, are discussed. Finally, the results are presented from a survey of primarily commercial MPS developers, that collected their views on the realistic responsibilities of each stakeholder group. The results also highlight their key perspectives on the use of MPS, in the context of Three Rs and regulatory applications.

Advances in RNA-Based Therapeutics: Challenges and Innovations in RNA Delivery Systems.

Nucleic acids, as carriers of genetic information, have found wide applications in both medical and research fields, including gene editing, disease diagnostics, and drug development. Among various types of nucleic acids, RNA offers greater versatility compared to DNA due to its single-stranded structure, ability to directly encode proteins, and high modifiability for targeted therapeutic and regulatory applications. Despite its promising potential in biomedicine, RNA-based medicine still faces several challenges. Notably, one of the most significant technical hurdles is achieving efficient and targeted RNA delivery while minimizing immune responses. Various strategies have been developed for RNA delivery, including viral vectors, virus-like particles (VLPs), lipid nanoparticles (LNPs), and extracellular vesicles (EVs). In this review, we explore the applications of these delivery methods, highlight their advantages and limitations, and discuss recent research advancements, providing insights for the future of RNA-based therapeutics.

A comprehensive review of atomically precise metal nanoclusters with emergent photophysical properties towards diverse applications.

Atomically precise metal nanoclusters (MNCs) composed of a few to hundreds of metal atoms represent an emerging class of nanomaterials with a precise composition. With the size approaching the Fermi wavelength of electrons, their energy levels are well-separated, leading to molecule-like properties, like discrete single electronic transitions, tunable photoluminescence (PL), inherent structural anisotropy, and distinct redox behavior. Extensive synthetic efforts and electronic structure revelation have expanded applicability of MNCs in catalysis, optoelectronics, and biology. This review highlights the intriguing photophysical and electrochemical behaviors of MNCs and their regulatory parameters and applications. Initially, we present a brief discussion on the evolution of MNCs from gas-phase naked metal clusters to monolayer ligand-protected MNCs along with representative studies on their electronic structure. Due to their quantized molecular orbitals, they often exhibit PL, which can be regulated based on their capping ligands, number of atoms, crystal packing, presence of heterometal, and surrounding environment. Apart from PL, the relaxation pathways of MNCs on an ultrafast time scale have been extensively studied, which significantly differ from that of plasmonic metal nanoparticles. Moreover, their interaction with high-intensity light results in unique non-linear optical properties. The synergy between MNCs in a hierarchical self-assembled structure has been exploited to enhance their PL by precisely tuning their non-covalent interactions. Moreover, several NC-based hybrids have been designed to exhibit efficient electron or energy transfer in the photoexcited state. In the next section, we briefly focus on the redox behavior of NCs and facile electron transfer to suitable substrates, which result in enzyme-like catalytic activity. Utilizing these photophysical and electrochemical behaviors, NCs are widely employed in catalysis, optical sensing, and light-harvesting applications, which are also discussed in this review. In the final section, conclusions and open questions for the NC research community are included. This review will provide a comprehensive view of the emerging physicochemical properties of MNCs, thereby enabling an understanding for their precise modulation in future.

Directors’ duties and AI regulation

ABSTRACT Companies are powerful initiators and adopters of AI, as they have been of previous industrial revolutions. Their boards of directors and the regulatory frameworks that govern directors will inevitably influence the development and implementation of AI. However, despite intense attention to AI regulation the regulatory contribution of directors’ duties remains relatively unexamined. This article critically analyses the capacity of these longstanding legal principles to assist with wider AI regulatory goals, demonstrating that directors’ duties have direct consequences for the regulation of AI. Evolved in response to the governance context of a complex artificial entity, the corporation, directors’ duties map onto key AI regulatory requirements. These include the need for broadly accepted but flexible and responsive norms, and a capacity to engage post-facto with both nuanced, complex fact situations and opaque decision making processes. Directors’ duties jurisprudence brings with it experience of reverse-engineering fact situations and responding to disembodied legal actors, two factors that may have wider AI regulatory application. Directors’ duties also demonstrate responsiveness to changing social perspectives, facilitating integration of developing attitudes to AI. Attending to the AI regulatory role played by directors’ duties can inform effective AI regulation and facilitate improved regulatory design overall.

Excessive Pharmaceutical Pricing and Repercussions on Paediatric Health Services

Context: Paediatric pharmaceutics have a significant share in expensive drugs. Recent years have seen significant calls for intervention against high prices for pharmaceutical products. The prohibition of prices that are excessive as a form of abuse of dominance is considered as Competition law violation in the European Union Law and national legal orders of its Member States. Majority of the Organization for Economic Cooperation and Development (OECD) member countries have similar prohibitions in their respective Competition laws with important exemption of United States (US) Antirust Law, although there is some indication of change in US exactly in the area of pharmaceutical sector. Regulation of excessive prices also exists in Competition laws of all BRICS (Brazil, Russian Federation, India, China, and South Africa) countries. The problem of excessive prices in pharmaceutics is one of the emerging themes in Competition law literature and case law development. This is the area where intellectual property rights, usually patent protection, collide with competition rules. Costs of research and development for such pharmaceutics are often very high emphasizing the need for balance between colliding legitimate interests: access to health and research development. Aim: To present regulatory and case law development of application of excessive prices as form of abuse of dominance in pharmaceutical sector with emphasis on paediatric health services. Data source: OECD, PubMed, Google Scholar, Scopus. Conclusion: Balance between competition rules on excessive prices as form of abuse of dominance and intellectual property rights in the area of expensive paediatric pharmaceutics can be regarded as suboptimal. Thus, there is a need for further regulatory development and application of rules on excessive prices in that field.

Genome-wide characterization of PEBP genes in Mung bean (Vigna radiata L.) with functional analysis of VrFT1 in relation to photoperiod

Mung bean (Vigna radiata L.), a widely cultivated legume, belongs to the Fabaceae family’s Papilionoideae subfamily. Although Phosphatidylethanolamine-binding protein (PEBP) genes have been identified in several plant species, their presence and function in mung bean remain largely unexplored. In this study, we identified seven VrPEBP genes from mung bean and classified them into four clades: FT, MFT, TFL and FT-like. Cis-regulatory element analysis revealed that VrPEBP genes may play a role in light, hormone, and stress responses. Quantitative real-time PCR (qRT-PCR) analysis indicated that VrPEBPs were constitutively expressed in various tissues. However, tissue-specific expression patterns were observed among VrPEBP genes. Under short-day (SD) conditions, VrFT1 and VrMFT1 exhibited significantly higher expression levels than under long-day (LD) conditions at 8 and 4 h, respectively. Conversely, VrTFL2 and VrTFL3 showed significantly higher expression levels under LD conditions compared to SD conditions at 8 and 12 h, respectively. The varied expression patterns of these genes under different photoperiod suggest their potential involvement in the photoperiodic regulation of flowering in mung bean. Additionally, phenotypic analysis of transgenic Arabidopsis plants overexpressing VrFT1 revealed higher expression levels under SD conditions and predicted its role in promoting flowering. These results provide valuable insights into the evolution and function of PEBP genes in mung bean and lay the foundation for further research on their regulatory mechanisms and potential applications in mung bean improvement.

Termination of Prosecution Based on Restorative Justice in Indonesia it is Associated with the Renewal Criminal Law

Criminal acts arise from various aspects of human activity, including the political, social and economic spheres. The success of law enforcement is traditionally measured by the ability to bring criminals to justice and impose appropriate penalties. However, in certain minor cases, such as theft, embezzlement, and fraud with minimal losses, the public questions the need for prosecution, as they believe that prosecution does not match the seriousness of the offense. The purpose of this study is to analyze the implications and effectiveness of prosecution discontinuation based on restorative justice in the Indonesian legal framework. Using a qualitative research approach, this study examines the regulatory structure and practical application of restorative justice in Indonesia. Data was collected through document analysis and interviews with legal practitioners to explore alignment of regulations with legal principles and community expectations. The research findings show that, although restorative justice offers a viable avenue to achieve peace between offenders and victims, its implementation in Indonesia is still limited, as it is not yet embedded in the broader criminal law system. Currently, peace agreements between the parties are only considered as a mitigating factor in the sentencing process. This research suggests that stronger legal legitimacy for restorative justice, such as the inclusion of restorative justice into the Draft Criminal Procedure Code is essential to adapt to the evolving legal culture and public expectations. The implications of this research highlight the need for comprehensive criminal law reform to ensure justice that aligns with societal values and contemporary legal standards.

Clinical Pharmacology of Asciminib: A Review.

Asciminib is a first-in-class allosteric inhibitor of the kinase activity of BCR::ABL1, specifically targeting the ABL myristoyl pocket (STAMP). This review focuses on the pharmacokinetic (PK) and pharmacodynamic data of asciminib, which is approved at a total daily dose of 80 mg for the treatment of adult patients with chronic myeloid leukemia in chronic phase who are either resistant or intolerant to ≥ 2 tyrosine kinase inhibitors or those harboring the T315I mutation (at a dose of 200 mg twice daily). Asciminib is predicted to be almost completely absorbed from the gut, with an absolute bioavailability (F) of approximately 73%. It should be administered in a fasted state, as food (particularly high-fat meals) reduces exposure. Asciminib displays a slightly greater than dose-proportional increase in exposure, with no time-dependent changes in PK observed following repeated dosing. This drug shows low clearance (6.31 L/h), with a moderate volume of distribution (111 L) and high human plasma protein binding (97.3%). The apparent terminal elimination half-life (t1/2) across studies was estimated to be between 7 and 15h. The PK of asciminib is not substantially affected by body weight, age, gender, race, or renal or hepatic impairment. Asciminib is primarily metabolized via CYP3A4-mediated oxidation (36.0%) and UGT2B7- and UGT2B17-mediated glucuronidation (13.3% and 7.8%, respectively); biliary secretion via breast cancer resistance protein contributes to about 31.1% to total systemic clearance, which is mainly through hepatic metabolism and biliary secretion through the fecal pathway, with renal excretion playing a minor role. The potential for PK drug interaction for asciminib both as a victim and a perpetrator has been summarized here based on clinical and predicted drug-drug interaction studies. Robust exposure-response models characterized asciminib exposure-efficacy and exposure-safety relationships. In patients without the T315I mutation, the exposure-efficacy analysis of the time course of BCR::ABL1IS percentages highlighted the existence of a slightly positive, albeit not clinically significant, relationship. Higher exposure was required for efficacy in patients harboring the T315I mutation compared with those who did not. The exposure-safety relationship analysis showed no apparent association between exposure and adverse events of interest over the broad range of exposure or dose levels investigated. Asciminib has also been shown to have no clinically relevant effect on cardiac repolarization. Here, we review the clinical pharmacology data available to date for asciminib that supported its clinical development program and regulatory applications.

Pharmacokinetics and absorption, distribution, metabolism andexcretion profiling of tanimilast following an intravenous 14C-microtracer coadministered with an inhaled dose in healthy male individuals.

Tanimilast is an inhaled phosphodiesterase-4 inhibitor currently in phase III clinical development for treating chronic obstructive pulmonary disease and asthma. This trial aimed to characterize the pharmacokinetics, mass balance, and metabolite profiling of tanimilast. Eight healthy male volunteers received a single dose of nonradiolabeled tanimilast via powder inhaler (Chiesi NEXThaler [3200 μg]), followed by a concomitant intravenous infusion of a microtracer ([14C]-tanimilast: 18.5 μg and 500 nCi). Plasma, whole blood, urine, and feces samples were collected up to 240 hours after dose to quantify nonradiolabeled tanimilast, [14C]-tanimilast, and total-[14C]. The inhaled absolute bioavailability of tanimilast was found to be approximately 50%. Following intravenous administration of [14C]-tanimilast, plasma clearance was 22L/h, the steady-state volume of distribution was 201 L, and the half-life was shorter compared to inhaled administration (14 vs 39 hours, respectively), suggesting that plasma elimination is limited by the absorption rate from the lungs. Seventy-nine percent (71% in feces; 8% in urine) of the intravenous dose was recovered in excreta as total-[14C]. [14C]-tanimilast was the major radioactive compound in plasma, whereas no recovery was observed in urine and only 0.3% was recovered in feces, indicating predominant elimination through metabolic route. Importantly, as far as no metabolites accounting for more than 10% of the circulating drug-related exposure in plasma or the administered dose in excreta were detected, no further qualification is required according to regulatory guidelines. This study design successfully characterized the absorption, distribution, and elimination of tanimilast, providing key pharmacokinetic parameters to support its clinical development and regulatory application. SIGNIFICANCE STATEMENT: This trial investigates pharmacokinetic and absorption, distribution, metabolism and excretion profile of tanimilast, an inhaled phosphodiesterase-4 inhibitor for chronic obstructive pulmonary disease and asthma. Eight male volunteers received a dose of nonradiolabeled tanimilast via Chiesi NEXThaler and a microtracer intravenous dose. Results show pivotal pharmacokinetic results for the characterization of tanimilast, excretion route and quantification of significant metabolites, facilitating streamlined clinical development and regulatory approval.

Modeling and Simulations in Latin-American Generic Markets: Perspectives from Chilean Local Industry, Regulatory Agency, and Academia.

In the last 20 years, modeling and simulations (M&S) have gained increased attention in pharmaceutical sciences. International industry and world-reference agencies have used M&S to make cost-efficient decisions through the model-informed drug development (MIDD) framework. Modeling tools include biopredictive dissolution models, physiologically based pharmacokinetic models (PBPK), biopharmaceutic models (PBBM), and virtual bioequivalence, among many others. Regulatorily, health agencies are becoming more and more open to accept the use of M&S to support regulatory applications, including setting dissolution specifications, quality-by-design (QbD), postapproval changes (SUPAC), etc. Nonetheless, the potential of M&S has been only barely explored in Latin America (Latam) across different actors: industry, regulatory agencies, and even academia. In this manuscript, we discuss the challenges and opportunities for implementing M&S approaches in Latam. Perspectives of regional experts were shared in a workshop. Attendance (professionals from industry, regulator, academia, and clinicians) also shared their views via survey. The rational development of bioequivalent generics was considered the main opportunity for M&S in regional market, particularly the use of PBPK and PBBM. Nonetheless, a critical mass of modeling scientists is needed before Latin American industry and regulators can actually benefit from M&S. Collaborations (e.g., Academia-Industry and Academia-Regulatory) may be a path to develop applied research projects and train the future modelers. Reaching that critical mass, scientists from industry may apply modeling across generic drug development process and life cycle, while regulatory scientists may issue guidelines in local language to support regional industry. Only at that stage could the full potential of MIDD be reached in Latin American generic markets.

Green cryptocurrency and business strategies: Framework and insights from a stewardship literature review

AbstractThis study critically evaluates the extant research on green cryptocurrency (GC). It incorporates the systematic literature review (SLR) approach of research executed through the analysis and compilation of 54 relevant studies. The focus is on identifying and interpreting the thematic foci and existing gaps about GC to inform potential areas of future research. The SLR findings aggregated the research around key themes: interconnectedness, portfolio diversification, environmental impact, green blockchain, behavioral impact, and regulatory policy. Additionally, the study develops the research profile of the selected studies in terms of data analysis methodology, research findings, variables investigated, and critical knowledge gaps in the literature corpus. The study proffers an integrated framework and actionable inferences for supporting the merits of novel research and regulatory project applications in GC. It is crucial for understanding and advancing research in the GC domain, which currently has scant academic literature and efforts toward sustainable investment.

Experiences and initiatives on pharmacokinetic modeling and simulation data analysis: Perspectives from the Brazilian Health Regulatory Agency (ANVISA)

The landscape of drug product development and regulatory sciences is evolving, driven by the increasing application of systems thinking and modeling and simulation (M&S) techniques, especially from a biopharmaceutics perspective. Patient-centric quality standards can be achieved within this context through the application of quality by design (QbD) principles and M&S, specifically by defining clinically relevant dissolution specifications (CRDS). To this end, it is essential to bridge in vitro results to drug product in vivo performance, emphasizing the need to explore the translational capacity of biopharmaceutics tools. Physiologically based M&S analyses offer a unique avenue for integrating the drug, drug product, and biological properties of a target organism to study their interactions on the pharmacokinetic response. Accordingly, Physiologically Based Biopharmaceutics Modeling (PBBM) has seen increasing use to support drug development and regulatory applications globally. In Brazil, a Model-Informed Drug Development (MIDD) policy and strategic project are not yet established, limiting applicability of M&S techniques. Drawing from the experience of the ANVISA-Academia PBBM Working Group (WG), this article assesses the opportunities and challenges for pharmacometrics (PMx) in Brazil and proposes strategies to advance the adoption of M&S analyses into regulatory decision-making.

Adipose tissue responds to stress-induced immunosuppression affecting immune response partially by miR-145-5p/S1PR1 pathway

Stress-induced immunosuppression (SIIS) is one of the most common problems in intensive poultry production, which can cause immunized chickens to still develop diseases and bring huge losses to production. Recently, adipose tissue, as an immunomodulatory organ, has become a hot topic of attention. However, the function and mechanism of adipose tissue involved in SIIS and its influence on the immune response are still unclear. In this study, we dynamically analyzed the correlations between the T cells migration and change of sphingosine-1-phosphate receptor 1(S1PR1) gene in adipose tissue using chicken models with different immune states, and further explored the regulatory mechanisms and application. The results showed that SIIS could significantly change the expressions of lymphocytes migration related S1PR1 gene, and SIIS could inhibit the Newcastle disease virus (NDV) immune response partially by affecting the migration and proliferation of TCRα+ T cells in adipose tissue. Moreover, the miR-145-5p/S1PR1 pathway was a potential key mechanism to regulate T cells migration in adipose tissue, and circulating miR-145-5p had potential value as a molecular marker. This research can provide innovative reference for in-depth studying the immunoregulatory function and mechanism of adipose tissue.

Investigating Data Protection Compliance Challenges

In today's landscape, safeguarding sensitive data is crucial for Organizations, but navigating data protection regulations and ensuring compliance is increasingly challenging. This research project explores Organizations' hurdles in achieving data protection compliance, offering insights to develop more effective strategies. A survey via Google Forms gathered insights from data protection experts and professionals, revealing key challenges such as difficulty understanding complex regulations, limited resources, and obstacles in implementing compliance measures. The study also reviewed the existing data protection regulatory framework and relevant literature, uncovering a common theme of confusion and a gap between regulatory requirements and practical application across Organizations. The research recognises that data protection extends beyond regulatory compliance, reflecting the evolving expectations of individuals and customers regarding the ethical handling of their data. This underscores the importance of data protection as both a legal and ethical responsibility closely tied to organisational reputation and public trust. The findings highlight the need for more precise, accessible guidelines and support mechanisms to bridge the gap between regulatory demands and organisational implementation. By addressing these challenges, Organizations can strengthen their data protection measures, foster trust, and ensure the security of sensitive information.

Genome-Wide Identification and Expression Analysis of the Cys2His2 Zinc Finger Protein Gene Family in Flammulina filiformis.

Zinc finger proteins (ZFPs) are essential transcription factors in eukaryotes, particularly the extensively studied C2H2 family, which is known for its involvement in various biological processes. This research provides a thorough examination and analysis of the C2H2-ZFP gene family in Flammulina filiformis. Using bioinformatics tools, 58 FfC2H2-ZFP genes spread across 11 chromosomes were identified and scrutinized in detail for their gene structures, protein characteristics, and phylogenetic relationships. The study of phylogenetics and synteny sheds light on the evolutionary relationships among C2H2-ZFPs in F. filiformis and other fungi, revealing a complex evolutionary past. The identification of conserved cis-regulatory elements in the gene promoter regions suggests intricate functionalities, particularly in the developmental and stress response pathways. By utilizing RNA-seq and qRT-PCR techniques, the expression patterns of these genes were explored across different developmental stages and tissues of F. filiformis, unveiling distinct expression profiles. Notably, significant expression variations were observed in the stipe elongation region and pilei of various sizes, indicating potential roles in fruiting body morphogenesis. This study enhances our knowledge of the C2H2-ZFP gene family in F. filiformis and lays the groundwork for future investigations into their regulatory mechanisms and applications in fungal biology and biotechnology.