Rational Design Approach Research Articles

In-Depth Structural Simplification of Celangulin V: Design, Synthesis, and Biological Activity.

Celangulin V is a novel botanical insecticide with significant bioactivity and a unique molecular target, but its complex polyol ester structure hinders its broader application in agriculture. To discover new analogues of celangulin V with a simpler structure and enhanced biological activities, we initiated a research project aimed at simplifying its structure and assessing insecticidal efficacy. In this study, a series of novel 1-tetralone derivatives were designed via a structure-based rational design approach and synthesized by a facile method. The biological activities of the target compounds were determined against Mythimna separata (M. separata), Plutella xylostella, and Rhopalosiphum padi. The results revealed that most of the synthesized compounds exhibited superior activities compared to celangulin V. Remarkably, the insecticidal activity of compound 6.16 demonstrated 102-fold greater stomach toxicity than celangulin V against M. separata. In addition, certain compounds showed significant contact toxicity against M. separata, a finding not reported previously in the structural optimization studies of celangulin V. Molecular docking analysis illustrated that the binding pocket of compound 6.16 with the H subunit of V-ATPase was the same as celangulin V. This study presents novel insights into the structural optimization of botanical pesticides.

Synthesis and evaluation of a self-standing CuBi2O4/CuO/Fe2O3 electrode for arsenic removal via photoelectrocatalysis

Water contamination by arsenic (As) poses a significant threat to millions of people worldwide. The development of an effective, affordable, and decentralized system for As removal remains a critical need, particularly to treat As at environmentally relevant concentrations. In this study, a self-standing electrode composed of an engineered interface of CuBi2O4/CuO/Fe2O3 was synthesized on fluorine-doped tin oxide (FTO) substrate using electrodeposition and dip-coating techniques. Each layer Bi2O3, CuBi2O4, CuO, and Fe2O3 was characterized photochemically and electrochemically to identify their light harvesting and electron transfer capacities. The efficacy of the CuBi2O4/CuO/Fe2O3 electrode for removing 0.5 mg L−1 As (III) was evaluated under various treatment conditions, including adsorption, photocatalysis (PC), electrocatalysis (EC), and photoelectrocatalysis (PEC). The PEC treatment at 1.0 V vs Ag/AgCl under visible light irradiation exhibited outstanding As removal efficiency of 96.8 %, surpassing PC (40.4 %) and EC (54.5 %). Fundamental evaluation of the removal mechanism revealed that CuBi2O4/CuO/Fe2O3 oxidizes As (III) to As (V) for enhanced adsorption onto iron oxide sites, as confirmed by X-ray photoelectron spectroscopy (XPS), which identified both As (III) and As (V) adsorbed on the surface. These findings underscore the importance of a rational design approach for photoanodes in As adsorption, highlighting the significant role of individual layers.

Point-of-care testing of Pseudomonas aeruginosa using PCN-222(Pt) prepared by nanoconfinement-guided protocol to catalyze gas generation reaction

BackgroundPathogenic bacteria are keeping threatening global public health since they can cause many infectious diseases. The traditional microorganism identification and molecular diagnostic techniques are insufficiently sensitive, time-consuming, or expensive. Thus it is of great interest to establish pressure signal-based sensing platforms for point-of-care testing of pathogenic bacteria to achieve timely diagnosis of infectious diseases. Rational design and synthesis of nano-sized probes with high peroxidase-mimicking activity have been a long-term cherished goal for improving the sensitivity of pressure signal-based sensing methods. ResultsGuided by nanoconfinement effect, PCN-222(Pt) was prepared by confining Pt clusters within the channels of a zirconium porphyrin MOFs material termed as PCN-222. In comparison to regular platinum nanoparticles, palladium@platinum core-shell nanodendrites, and platinum-coated gold nanoparticles, the prepared PCN-222(Pt) displayed superior peroxidase-mimicking activity with outstanding efficiency for catalyzing the decay of H2O2 to produce O2. Thus it was used as a pressure signal probe to establish a sensitive method on a hydrogel pellets platform for analyzing Pseudomonas aeruginosa (P. aeruginosa), for which polymyxin B and a phage termed as JZ1 were used as recognition agents for the target pathogen. P. aeruginosa was quantified with a handheld pressure meter within a broad range of 2.2 × 102–2.2 × 107 cfu mL−1. This method was used to quantify P. aeruginosa in various biological and food samples with acceptable accuracy and reliability. SignificanceThe proposed nanoconfinement-guided protocol provides a novel approach for rational design and preparation of nano-sized probes with high peroxidase-mimicking activity for catalyzing gas-generation reaction. Thus this study opens an avenue for establishment of sensitive pressure signal-based sensing methods for pathogenic bacteria, which shows broad application prospects in medical diagnosis of infectious diseases.

Constructing a Built-In Electric Field To Accelerate Water Dissociation for Efficient Alkaline Hydrogen Evolution.

The alkaline hydrogen evolution reaction (HER) is intricately linked to the water dissociation kinetics. The quest for new strategies to accelerate this step is a pivotal aspect of enhancing the HER performance. Herein, we designed and synthesized a heterogeneous nickel phosphide/cobalt phosphide nanowire array grown on nickel foam (Ni2P/CoP/NF) to form a p-n junction structure. The built-in electric field (BEF) in the p-n junction optimizes the binding ability of hydrogen and hydroxyl intermediates, efficiently promoting water dissociation for the alkaline HER. Consequently, Ni2P/CoP/NF exhibits a lower overpotential of 58 and 118 mV at 30 and 100 mA cm-2, respectively, and high stability over 40 h at 300 mA cm-2 for the HER in 1 M KOH. Computational calculations combined with experiment results testify that the BEF presence in the p-n junction of Ni2P/CoP/NF effectively promotes water dissociation, regulates intermediate adsorption/desorption, and boosts electron transport. This study presents a rational design approach for high-performance heterogeneous electrocatalysts.

Unlocking the Charge-Storage Potential of a Phenanthraquinone-based Two-Dimensional Covalent Organic Framework (2D COF).

The high surface area, open pore-structure and atomic-level organization inherent in many covalent organic frameworks (COFs) make them an attractive polymer platform for developing functional materials. Herein, a chemically robust 2D COF (TpOMe-DAPQ COF) containing phenanthraquinone moieties was prepared by condensing 2,4,6-trimethoxy-1,3,5-benzenetricarbaldehyde (TpOMe) and 2,7-diamino-9,10-phenanthraquinone (DAPQ) using the convenient mechanochemical method. The poor charge-storage capacity of the pristine TpOMe-DAPQ COF was substantially improved by first investigating its redox-site accessibility (RSA) using different conductivity-enhancement methods, and then optimizing the amount of EDOT needed to perform an in-situ polymerization. The resulting composite (0.4EDOT@TpOMe-DAPQ) was characterized and its enhanced charge-storage capabilities enabled it to be used as an anode material in an aqueous Mn beaker-cell battery capable of delivering 0.76 V. This work outlines the rational design approach used to develop a functional charge-storage material utilizing a COF-based polymerization platform.

Rational design of Bacteria-Targeted and Photo-Responsive MOF gel with antibacterial and anti-inflammatory function for infected wound healing

Damaged skin is prone to recurrent bacterial infections, which hinder proper wound healing and exacerbate inflammation. Urgent attention is required to devise versatile dressings possessing antibacterial and anti-inflammatory attributes to facilitate wound recovery. In this study, we engineered a photo-responsive metal–organic frameworks (MOFs) gel, targeting bacteria, and endowed it with antibacterial and anti-inflammatory effects to address infected wound healing. On the one hand, the anionic MOF was choose to support cationic natural drug Berberine (BBR) to achieve slow release of anti-inflammatory drugs. On the other hand, the incorporation of Ag nanoparticles onto MOFs not only facilitates the generation of reactive oxygen species (ROS) via photocatalysis under visible light but also furnishes sites for boric acid group attachment to target bacteria and expedites ROS transfer. The resultant MOFs@Ag-B@BBR exhibited potent photodynamic antibacterial activity against Staphylococcus aureus and MRSA at a low concentration of 37.5 µg mL−1, surpassing the efficacy of most reported MOFs-based nano-antimicrobials. Upon application of the MOFs@Ag-B@BBR composite hydrogel to infected wounds, the released BBR, in conjunction with photodynamic antibacterial action, significantly mitigated inflammatory responses and fostered wound healing. This study provides a widely applicable approach for rational design of MOFs materials with enhance antibacterial and anti-inflammatory functions.

Hepatitis C Virus E1E2 Structure, Diversity, and Implications for Vaccine Development.

Hepatitis C virus (HCV) is a major medical health burden and the leading cause of chronic liver disease and cancer worldwide. More than 58 million people are chronically infected with HCV, with 1.5 million new infections occurring each year. An effective HCV vaccine is a major public health and medical need as recognized by the World Health Organization. However, due to the high variability of the virus and its ability to escape the immune response, HCV rapidly accumulates mutations, making vaccine development a formidable challenge. An effective vaccine must elicit broadly neutralizing antibodies (bnAbs) in a consistent fashion. After decades of studies from basic research through clinical development, the antigen of choice is considered the E1E2 envelope glycoprotein due to conserved, broadly neutralizing antigenic domains located in the constituent subunits of E1, E2, and the E1E2 heterodimeric complex itself. The challenge has been elicitation of robust humoral and cellular responses leading to broad virus neutralization due to the relatively low immunogenicity of this antigen. In view of this challenge, structure-based vaccine design approaches to stabilize key antigenic domains have been hampered due to the lack of E1E2 atomic-level resolution structures to guide them. Another challenge has been the development of a delivery platform in which a multivalent form of the antigen can be presented in order to elicit a more robust anti-HCV immune response. Recent nanoparticle vaccines are gaining prominence in the field due to their ability to facilitate a controlled multivalent presentation and trafficking to lymph nodes, where they can interact with both the cellular and humoral components of the immune system. This review focuses on recent advances in understanding the E1E2 heterodimeric structure to facilitate a rational design approach and the potential for development of a multivalent nanoparticle-based HCV E1E2 vaccine. Both aspects are considered important in the development of an effective HCV vaccine that can effectively address viral diversity and escape.

Exploring the unexplored chemical space: Rational identification of new Tafenoquine analogs with antimalarial properties

Patents tend to define a huge chemical space described by the combinatorial nature of Markush structures. However, the optimization of new principal active ingredient is frequently driven by a simple Free Wilson approach. This procedure leads to a highly focused study on the chemical space near a hit compound leaving many unexplored regions that may present highly biological active reservoirs. This study aims to demonstrate that this unveiled chemical space can hide compounds with interesting potential biological activity that would be worth pursuing. This underlines the value and necessity of broadening an approach beyond conventional strategies. Hence, we advocate for an alternative methodology that may be more efficient in the early drug discovery stages. We have selected the case of Tafenoquine, a single-dose treatment for the radical cure of P. vivax malaria approved by the FDA in 2018, as an example to illustrate the process. Through the deep exploration of the Tafenoquine chemical space, seven compounds with potential antimalarial activity have been rationally identified and synthesized. This small set is representative of the chemical diversity unexplored by the 58 analogs reported to date. After biological assessment, results evidence that our approach for rational design has proven to be a very efficient exploratory methodology suitable for the early drug discovery stages.

Side‐Chain Engineering of Non‐Fullerene Acceptors with Trialkylsilyloxy Groups for Enhanced Photovoltaic Performance†

Comprehensive SummarySide‐chain engineering has emerged as a highly effective strategy for tailoring the aggregation behavior and charge transport properties of non‐fullerene small molecule acceptors (SMAs). In this study, we designed and synthesized two SMAs, namely BTPSi‐Bu and BTPSi‐Pr, respectively incorporating tributylsilyloxy and triisopropylsilyloxy groups in their outer positions. Notably, BTPSi‐Bu exhibited better planarity, crystallization, and favorable phase separation when paired with PM6 donor polymer compared to its counterpart, BTPSi‐Pr. The resulting organic solar cells, utilizing the PM6:BTPSi‐Bu blend, demonstrated a remarkable power conversion efficiency of 17.41% and a high open‐circuit voltage of 0.859 V. These findings underscore the significance of integrating trialkylsilyloxy side chains into SMAs as a rational design approach for enhancing the performance of photovoltaic systems.

Enhancement of rAAV titers via inhibition of the interferon signaling cascade in transfected HEK293 suspension cultures.

The production of recombinant adeno-associated virus (rAAV) for gene therapy applications relies on the use of various host cell lines, with suspension-grown HEK293 cells being the preferred expression system due to their satisfactory rAAV yields in transient transfections. As the field of gene therapy continues to expand, there is a growing demand for efficient rAAV production, which has prompted efforts to optimize HEK293 cell line productivity through engineering. In contrast to other cell lines like CHO cells, the transcriptome of HEK293 cells during rAAV production has remained largely unexplored in terms of identifying molecular components that can enhance yields. In our previous research, we analyzed global regulatory pathways and mRNA expression patterns associated with increased rAAV production in HEK293 cells. Our data revealed substantial variations in the expression patterns between cell lines with low (LP) and high-production (HP) rates. Moving to a deeper layer for a more detailed analysis of inflammation-related transcriptome data, we detected an increased expression of interferon-related genes in low-producing cell lines. Following upon these results, we investigated the use of Ruxolitinib, an interferon pathway inhibitor, during the transient production of rAAV in HEK293 cells as potential media additive to boost rAAV titers. Indeed, we find a two-fold increase in rAAV titers compared to the control when the interferon pathways were inhibited. In essence, this work offers a rational design approach for optimization of HEK293 cell line productivity and potential engineering targets, ultimately paving the way for more cost-efficient and readily available gene therapies for patients.

Rational Design of NIR-II G-Quadruplex Fluorescent Probes for Accurate In Vivo Tumor Metastasis Imaging.

Accurate in vivo imaging of G-quadruplexes (G4) is critical for understanding the emergence and progression of G4-associated diseases like cancer. However, existing in vivo G4 fluorescent probes primarily operate within the near-infrared region (NIR-I), which limits their application accuracy due to the short emission wavelength. The transition to second near-infrared (NIR-II) fluorescent imaging has been of significant interest, as it offers reduced autofluorescence and deeper tissue penetration, thereby facilitating more accurate in vivo imaging. Nonetheless, the advancement of NIR-II G4 probes has been impeded by the absence of effective probe design strategies. Herein, through a "step-by-step" rational design approach, we have successfully developed NIRG-2, the first small-molecule fluorescent probe with NIR-II emission tailored for in vivo G4 detection. Molecular docking calculations reveal that NIRG-2 forms stable hydrogen bonds and strong π-π interactions with G4 structures, which effectively inhibit twisted intramolecular charge transfer (TICT) and, thereby, selectively illuminate G4 structures. Due to its NIR-II emission (940 nm), large Stokes shift (90 nm), and high selectivity, NIRG-2 offers up to 47-fold fluorescence enhancement and a tissue imaging depth of 5 mm for in vivo G4 detection, significantly outperforming existing G4 probes. Utilizing NIRG-2, we have, for the first time, achieved high-contrast visualization of tumor metastasis through lymph nodes and precise tumor resection. Furthermore, NIRG-2 proves to be highly effective and reliable in evaluating surgical and drug treatment efficacy in cancer lymphatic metastasis models. We are optimistic that this study not only provides a crucial molecular tool for an in-depth understanding of G4-related diseases in vivo but also marks a promising strategy for the development of clinical NIR-II G4-activated probes.

In situ patterning of nickel/sulfur-codoped laser-induced graphene electrode for selective electrocatalytic valorization of glycerol

The electrocatalytic valorization of glycerol is an emerging upcycling process for the biomass utilization. In this work, nickel/sulfur-codoped laser-induced graphene (LINiSG) on flexible polyimide film was fabricated and demonstrated as a self-supported and binder-free monolithic electrode for electrocatalytic glycerol oxidation reaction (EGOR) towards the selective generation of value-added three-carbon (C3) chemicals. The laser-induced in-situ and synchronous carbonization of polysulfone and Ni precursor was demonstrated to remarkably promote both the electrochemical surface area and Ni loading of LINiSG electrode. These synergistic merits are considered as the main origin for its enhanced EGOR activity and stability, showing much smaller Tafel slope and 2-fold enhanced current density, as compared to LINiG electrode without the co-carbonization of polysulfone. The influence of experimental parameters has been systematically investigated by the product analysis, achieving an average 94.7 % C3 selectivity and 76.1 % C3 Faradaic efficiency with a high selectivity of dihydroxyacetone (ca. 67.7 %) in near-neutral conditions, which is better than most of the reported EGOR electrodes operated in near-neutral conditions. This work demonstrated the viability of using flexible self-supported and binder-free LIG electrode with rational design and preparation approach for electrocatalytic valorization of biomass.

A rational approach for the scantling design of GRP pleasure crafts

In various fields of industrial engineering, including naval architecture, structural design is considered one of the most challenging steps by designers as it shall accommodate all other design aspects, likely already defined in earlier steps. Moreover, structures need to comply with specific requirements, i.e. limit states, while also performing in accordance with user's demands in terms of mission profile. In a goal-based design perspective, limit state criteria have to be defined accounting for both, structural robustness, functional and operational demands. Construction feasibility is an issue as well, further constraining the scantling design. Balancing construction needs, ship performances and structural aspects is not always an easy task.The environment in which a ship operates is indeed complex and verification of safety and robustness is usually carried out in accordance with classification societies and other rules, possibly applying direct calculations whenever applicable rules do not provide suitable solutions. At the same time, the asset needs to fulfill various attributes specified by the owner. As a matter of facts, the scantling design is mathematically a multivariate and over-constrained problem whose feasible solutions are in fact rather few. In the case of composite pleasure craft hull structures, while the number of design variables increases, feasibility constraints also largely reduce the design space.To address these challenges, designers may nowadays exploit new procedures able to reduce structural weight, though ensuring that the asset performs safely and adequately meeting client's requirements. To achieve this, designers may combine a more rational design philosophy with currently available numerical problem-solving applications to obtain a rational trade-off among all actually feasible solutions, which are relatively few.Thus, the objective of this study is to present a dependable numerical method for determining the scantling while reducing the weight of hull composite structures of pleasure crafts. This method aims to provide the most suitable, high-performing, and safe layout in the quickest and easiest possible manner. It turns out that counterintuitive, yet feasible and fit-for-purpose, structural lay-outs can be obtained.

Development of the Squaramide Scaffold for High Potential and Multielectron Catholytes for Use in Redox Flow Batteries.

Nonaqueous organic redox flow batteries (N-ORFBs) are a promising technology for grid-scale storage of energy generated from intermittent renewable sources. Their primary benefit over traditional aqueous RFBs is the wide electrochemical stability window of organic solvents, but the design of catholyte materials, which can exploit the upper range of this window, has proven challenging. We report herein a new class of N-ORFB catholytes in the form of squaric acid quinoxaline (SQX) and squaric acid amide (SQA) materials. Mechanistic investigation of decomposition in battery-relevant conditions via NMR, HRMS, and electrochemical methods enabled a rational design approach to optimizing these scaffolds. Three lead compounds were developed: a highly stable one-electron SQX material with an oxidation potential of 0.51 V vs Fc/Fc+ that maintained 99% of peak capacity after 102 cycles (51 h) when incorporated into a 1.58 V flow battery; a high-potential one-electron SQA material with an oxidation potential of 0.81 V vs Fc/Fc+ that demonstrated negligible loss of redox active material as measured by pre- and postcycling CV peak currents when incorporated in a 1.63 V flow battery for 110 cycles over 29 h; and a proof-of-concept two-electron SQA catholyte material with oxidation potentials of 0.48 and 0.85 V vs Fc/Fc+ that demonstrated a capacity fade of just 0.56% per hour during static H-cell cycling. These findings expand the previously reported space of high-potential catholyte materials and showcase the power of mechanistically informed synthetic design for N-ORFB materials development.

Integrated design optimization method for pavement structure and materials based on further development of finite element and particle swarm optimization algorithm

The maintenance design for existing asphalt pavements, characterized by their typical structure and material composition, falls short of meeting the diverse requirements of pavements. There is an urgent need for a more rational design approach that can achieve accurate pavement design while also reducing construction costs. This paper focuses on selected conventional road surface structures and conducts an analysis of how pavement structure and materials impact fatigue life and construction expenses. It employs Python to perform three-dimensional finite element road modeling, analysis, and data extraction at specific locations, using normalized fatigue equation to calculate pavement fatigue life. The optimization variables selected are the modulus E and thickness h, with pavement maintenance cost serving as the optimization target function. The particle swarm optimization algorithm is employed for optimizing these variables. A comparative analysis is carried out for five improved optimization algorithms, optimizing combinations of inertia weight, learning factors, particle speed, number of particles, and iteration times. The results reveal that the enhanced optimization algorithms significantly improve iteration trends, featuring velocity limitations and natural selection mechanisms. Between 0 and 20 iterations, all algorithms search for the optimal solution over a wider interval with greater fluctuations. Between 20 and 30 iterations, each algorithm gradually converges towards its solution, stabilizing in the later stages. Based on optimization speed and actual engineering cost benchmarks, the NDPSO algorithm is selected, resulting in a 52.8% reduction in actual engineering costs. Adjustments to the particle count and iteration number of the NDPSO algorithm are made based on computational cost and optimization accuracy, thus choosing 20 particles and 50 iterations as the recommended parameter combination. By combining automated finite element modeling, analysis, and particle swarm optimization, this paper offers a novel and effective automated design reference for extending the lifespan of asphalt pavement structures.

Nanorod-like Bimetallic Oxide for Enhancing the Performance of Supercapacitor Electrodes.

Supercapacitors are widely used in many fields owing to their advantages, such as high power, good cycle performance, and fast charging speed. Among the many metal-oxide cathode materials reported for supercapacitors, NiMoO4 is currently the most promising electrode material for high-specific-energy supercapacitors. We have employed a rational design approach to create a nanorod-like NiMoO4 structure, which serves as a conductive scaffold for supercapacitors; the straightforward layout has led to outstanding results, with nanorod-shaped NiMoO4 exhibiting a remarkable capacity of 424.8 F g-1 at 1 A g-1 and an impressive stability of 80.2% capacity preservation even after 3500 cycles, which surpasses those of the majority of previously reported NiMoO4 materials. NiMoO4//AC supercapacitors demonstrate a remarkable energy density of 46.31 W h kg-1 and a power density of 0.75 kW kg-1. This synthesis strategy provides a facile method for the fabrication of bimetallic oxide materials for high-performance supercapacitors.

Graphene aerogel encapsulated double carbon shell CoFe@C@C nanocubes for construction of high performance microwave absorbing materials

Recently, Prussian blue analogs (PBAs) have attracted much attention due to their transformation into dielectric/magnetic coupled composites after pyrolysis at high temperatures and their ability to maintain their basic morphology. However, designing efficient microwave-absorbing materials using PBAs is still a challenge. In this work, Co–Fe PBA was prepared by wet chemical method and graphene aerogel-encapsulated double carbon-shell CoFe@C@C nanocubes (CFCCG) were prepared by stirred self-polymerization and hydrothermal method, and heat-treated at different temperatures to obtain composites consisting of core-shell-structured PBA-derived particles encapsulated in a double layer of graphene and polydopamine (PDA). The addition of graphene not only reduces the density of the derived material but also effectively disperses the CoFe@C@C nanocubes to prevent the generated highly conductive carbon layer from adhering to improve the impedance matching, as well as introduces a large number of heterogeneous interfaces to improve the loss capability. Thanks to which the CFCCG7 composites have a reflection loss of −66.33 dB and an effective microwave absorption frequency covering the entire Ku-band. This work provides a new approach for rational design of multilayer carbon structure-coated PBA-based microwave absorbing materials.

Abstract 4076: Interleukin-18 engineered for resistance to IL-18 binding protein (IL-18BP) and half-life extension to enhance its therapeutic potential

Abstract Interleukin-18 (IL-18) possesses a unique combination of innate and acquired immune functions with high potential to be transformative in cancer immunotherapy of solid tumors. In clinical studies, recombinant wild-type IL-18 was limited by rapid upregulation of IL-18 binding protein (IL-18BP), which serves as a negative feedback checkpoint of the IL-18 pathway. As its name implies, IL-18BP tightly binds to IL-18, thereby blocking its ability to activate the IL-18 receptor. We have focused our protein engineering efforts to obtain variants of IL-18 that are resistant to IL-18BP suppression while retaining IL-18 biological activity. To further capitalize on the immunological effects of IL-18, the IL-18BP resistant variants were fused to half-life extension protein scaffolds for enhanced in vivo exposure. Rational protein design and combinatorial approaches were used to generate a pool of IL-18 variants that were screened for potency and resistance to IL-18BP. Several IL-18 variants showed undetectable binding to IL-18BP (up to 1 mM IL-18BP) and displayed a spectrum of potencies relative to recombinant wild-type IL-18. Importantly, these variants retained their full biological activity in the presence of super-physiological levels of IL-18BP, as tested with in vitro cell-based IFN-γ release assays. In vivo pharmacokinetic studies in mice compared several IL-18BP resistant variants fused to a half-life extension scaffold with a non-half-life enhanced IL-18BP resistant variant. The half-lives for the fusion proteins ranged from 15 - 30h, while the non-fused variant showed a half-live of 0.6h. Furthermore, the IL-18 variants displayed a more durable, Th1 immune response compared to a non-half-life enhanced variant. Thus, IL-18 variants with strong resistance to IL-18BP that are fused to a half-life extension protein scaffold displayed enhanced pharmacokinetic and pharmacodynamic properties in preclinical mouse models. These protein engineering modifications will enable us to select an appropriate development candidate with enhanced pharmaceutical properties to achieve the full therapeutic potential of IL-18 to assess in patients with cancer. Citation Format: Jean Chamoun, Clifford DiLea, Kristiana Dreaden, Pinar Gurel, Joshua Heiber, Robert G. Newman, Su-Ping Pearson, Chunhua Wang, Yanchun Zhao, Mark Whitmore. Interleukin-18 engineered for resistance to IL-18 binding protein (IL-18BP) and half-life extension to enhance its therapeutic potential [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4076.

Abstract 4066: Generation of tumor targeted self-assembling split IL-12 subunits for the treatment of cancer

Abstract A major milestone in immuno-oncology was the development of immune checkpoint inhibitors, resulting in significant benefits for numerous patients. However, many patients are not responsive, become refractory, or cannot tolerate the treatment due to toxicity or side effects, demonstrating the need for additional immunotherapies. Cytokines are a class of promising immunomodulatory proteins being explored as therapeutics, but their success has been limited due to their rapid clearance or pleiotropic properties. Classically known as ‘signal 3’ of an endogenous immune response, IL-12p70 is a heterodimeric cytokine comprising p35 and p40 subunits. When administered exogenously, IL-12 is a potent stimulator of the immune system and can have profound anti-tumor activity; however, the clinical use of IL-12 has been limited due to poor systemic tolerability. Therefore, developing an immunotherapy that can be systemically administered and improve the therapeutic index could capitalize on the potential clinical benefit of IL-12. Mural Oncology has developed an innovative approach to mitigate the toxicity of IL-12 by splitting the heterodimer into inactive p35 and p40 monomers. The individual subunits are separately fused to two non-competitive antibody fragments targeting a highly expressed tumor-associated antigen. The goal of this approach is to conditionally activate IL-12 preferentially in the tumor microenvironment (TME). This is achieved by sequential administration of the targeted subunits, which will drive assembly and activity of the IL-12 heterodimer primarily at the tumor site, reducing systemic exposure and thereby potentially reducing associated toxicities. To engineer the split IL-12 subunits, a structure-based rational design approach was used to remove the natural intra-molecular disulfide bond and generate variants that maintained affinity between the p35 and p40 monomers. The engineered subunits were characterized using analytical methods and screened for the pair with the strongest affinity. Additionally, the candidate pair was assessed for receptor binding and functional activity on human T cells through phosphorylation of STAT4 and secretion of IFN-γ. Pharmacokinetics and pharmacodynamics were evaluated in mouse models with and without tumors demonstrating that dual targeting led to an increase in tumor retention, rapid clearance from circulation, and dose dependent release of IFN-γ. The results thus far demonstrate that the innovative approach of tumor targeted self-assembling split IL-12 has the potential to enhance the overall therapeutic potential of this cytokine. A development candidate will be identified and moved into clinical testing in cancer patients to enable us to fully understand if this design approach holds true in the clinical setting. Citation Format: Joshua Heiber, Pinar Gurel, Robert G. Newman, Kristiana Dreaden, Yanchun Zhao, Chunhua Wang, Su-Ping Pearson, Jean Chamoun. Generation of tumor targeted self-assembling split IL-12 subunits for the treatment of cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4066.

Biological New Targets Prediction & ADME Pharmacokinetics Profiling of Newly Synthesized E / Z Isomers of Methyl 2-phenyl-2-(2-phenylhydrazono) Acetate Derivatives

Objective: In order to predict the pharmacokinetic properties, and new potential biological targets, biological targets prediction, and absorption, distribution, metabolism and excretion (ADME) profiling of newly synthesized Entgegen / Zusammen (E / Z isomers) of Methyl 2-phenyl-2-(2-phenylhydrazono) acetate derivatives were carried out. Methods: The prediction of new biological targets, pharmacokinetic properties, membrane permeability, and bioavailability radar properties were carried out by using Swiss Targets Prediction and Swiss ADME tools, respectively. Results: All the eight screened compounds possessed excellent drug-likeness criteria and passed successfully from Pan-assay interference compounds filter. Additionally, all screened compounds fell within the acceptable range in the bioavailability radar, showing excellent descriptors with a slight exceeding of insaturation properties. Compounds displayed elevated permeability across the Blood Brain Barrier, while compounds exhibited highest Human Intestinal Absorption according to the Egan Egg model. Moreover, the screened compounds also exhibited good pharmacokinetic properties. All the compounds exhibited activity against various enzymes, such as lyase, kinase, protease, and Family AG protein-coupled receptor. Conclusion: This conducted study smartly reveals that in-silico based studies were considered to provide robustness towards a rational drug design and development approach; therefore, in this way, it helps for further investigation of drug design and drug discovery steps.