Experimental Drug Research Articles

Effect of ascorbic acid on cellular respiration with mitochondrial reductase in gingival fibroblast.

Vitamin C or L-ascorbic acid has diverse functions in the body, especially healing promotion in tissue injury via participating in the hydroxylation reactions required for collagen formation. Systemic administration of vitamin C plays an important role on gingival fibroblast proliferation and functions. Whether local or rinsing administration of vitamin C alters gingival fibroblast wound healing behavior remains unclear. Therefore, it is of interest to investigate the effect of vitamin C on gingival fibroblast behavior utilizing cell culture. Primary human gingival fibroblasts isolated from gingival tissue were rinsed with medium containing various concentrations of vitamin C. Cell migration, cell viability was assessed using MTT assay. The viability assay showed >95% live cells, and no significant (P > 0.05) difference in these values was observed at different concentrations at 24 hrs. The levels of cell proliferation were not significantly different among the control and experimental groups in 24 hrs experimental time-points (p > 0.05). Vitamin C is nontoxic and can be used for further experiments to validate for clinical application. This was further confirmed with morphological examination after 24hrs incubation on control and experimental group drugs observed under the phase contrast microscope.

Clinical and pharmacological effectiveness of Fit-O-Metrin drug in the scheme of treatment of bovine postpartum metritis

The article presents data on the approval of the new Fit-O-Metrin drug for accelerating the recovery of cows from postpartum metritis. The experiment was focused on monitoring the excretion of exudate; restoration and strengthening of the contractile function and regenerative processes in the uterus after the use of the drug. Favorable conditions for uterine disease are often created during calving. However, the reproductive function of animals is mostly determined by the ability to restore the physiological state of the uterus in the postpartum period. Thus, in the experimental farm AgroTsvit 2012 (Zhytomyr region), clinical and laboratory examination of cows in the postpartum period determined that the prevalence of metritis covers up to 42% of the herd. The largest percentage of sick cows had purulent form (53.1 %), catarrhal-purulent metritis was registered in 30.4 %, catarrhal – in 16.5 %. Through our own research, we determined the effectiveness of the experimental drug Fit-O-Metrin, made on the basis of plant materials (extract of thyme, motherwort, and common barberry) for intrauterine administration as part of the treatment regimen for bovine metritis. In the clinical test, 13 sick cows were involved, to which Fit-O-Metrin was applied according to three different schemes. Another 5 animals with metritis were included in the control group, which were treated according to the standard protocol for the farm. However, regardless of the chosen scheme, the complete cessation of the release of inflammatory exudate from the vagina of the entire cow's herd was recorded already on the 6–7 day (in the control group – on the 9 day). Normalization of tone and anatomical position of the uterus, complete closure of the cervical canal was noted on the 8th day (in the control group – on the 12 day). The most optimal scheme of using the herbal preparation against postpartum purulent metritis is 2 days injections, 2 days break, 2 days injections, 2 days break, final injection. A single dose of the FitO-Metrin drug is 30 ml. Key words: postpartum metritis, Fit-O-Methrin, cattle, treatment scheme, effectiveness, exudate, sexual cycle.

Neuropathology of Anti-Amyloid-β Immunotherapy: A Case Report.

Host responses to anti-amyloid-β (Aβ) antibody therapy are evident in neuroimaging changes and clinical symptoms in a subset of clinical trial subjects receiving such therapy. The pathological basis for the imaging changes and clinical symptoms is not known, nor is the precise mechanism of Aβ clearing. We report the autopsy findings in a 65-year-old woman who received three open label infusions of the experimental anti-Aβ drug lecanemab over about one month. Four days after the last infusion, she was treated with tissue plasminogen activator for acute stroke symptoms and died several days later with multifocal hemorrhage. Neuropathological examination demonstrated histiocytic vasculitis involving blood vessels with cerebral amyloid angiopathy. Fragmentation and phagocytosis of vascular Aβ were present throughout the cerebral cortex. Phagocytosis of parenchymal Aβ plaques was noted. Changes suggestive of Aβ and phosphorylated tau "clearing" were also noted. The findings overall suggest that anti-Aβ treatment stimulated a host response to Aβ, i.e., target engagement. The findings also provide evidence that amyloid-related imaging abnormalities might be indicative of an Aβ phagocytic syndrome within cerebral vasculature and parenchymal brain tissue in some cases.

Repurposing Drugs for the Treatment of COVID-19 and Its Cardiovascular Manifestations.

COVID-19 is an infectious disease caused by SARS-CoV-2 leading to the ongoing global pandemic. Infected patients developed a range of respiratory symptoms, including respiratory failure, as well as other extrapulmonary complications. Multiple comorbidities, including hypertension, diabetes, cardiovascular diseases, and chronic kidney diseases, are associated with the severity and increased mortality of COVID-19. SARS-CoV-2 infection also causes a range of cardiovascular complications, including myocarditis, myocardial injury, heart failure, arrhythmias, acute coronary syndrome, and venous thromboembolism. Although a variety of methods have been developed and many clinical trials have been launched for drug repositioning for COVID-19, treatments that consider cardiovascular manifestations and cardiovascular disease comorbidities specifically are limited. In this review, we summarize recent advances in drug repositioning for COVID-19, including experimental drug repositioning, high-throughput drug screening, omics data-based, and network medicine-based computational drug repositioning, with particular attention on those drug treatments that consider cardiovascular manifestations of COVID-19. We discuss prospective opportunities and potential methods for repurposing drugs to treat cardiovascular complications of COVID-19.

Chemical sphincterotomy in posthemorrhoidectomy pain relief: a meta-analysis

PurposeThis study aims to evaluate the pain relief function of chemical sphincterotomy in patients undergoing haemorrhoid surgery and compare, through a meta-analysis, the different drugs used to treat this condition.MethodsWe conducted a search in databases including PubMed, EMBASE and Web of Science. The methodological quality was evaluated using the Revised Cochrane risk-of-bias tool for randomized trials (ROB2). The pain score was assessed using a visual analogue scale (VAS) on day 1, day 2, and day 7, and a meta-analysis was conducted based on the use of random effects models. In addition, the subgroup analysis was evaluated based on the kind of experimental drugs. Heterogeneity and publication bias were assessed.ResultsFourteen studies with a total of 681 patients were included in this meta-analysis, and all studies were randomized controlled trials RCTs. Chemical sphincterotomy showed better pain relief function than placebo on day 1 (SMD: 1.16, 95% CI 0.52 to 1.80), day 2 (SMD: 2.12, 95% CI 1.37 to 2.87) and day 7 (SMD: 1.97, 95% CI 1.17 to 2.77) after surgery. In the subgroup meta-analysis, we found that different drugs for chemical sphincterotomy provided different pain relief.ConclusionChemical sphincterotomy effectively relieves pain after haemorrhoidectomy, and calcium channel blockers have the best effect.

7-Chloroquinolinehydrazones as First-in-Class Anticancer Experimental Drugs in the NCI-60 Screen among Different Investigated Series of Aryl, Quinoline, Pyridine, Benzothiazole and Imidazolehydrazones.

In the context of a continuously increasing global cancer risk, the search for new effective and affordable anticancer drugs remains a constant demand. This study describes chemical experimental drugs able to destroy cancer cells by arresting their growth. New hydrazones with quinoline, pyridine, benzothiazole and imidazole moieties have been synthesized and evaluated for their cytotoxic potential against 60 cancer cell lines. 7-Chloroquinolinehydrazones were the most active in the current study and exhibited good cytotoxic activity with submicromolar GI50 values on a large panel of cell lines from nine tumor types (leukemia, non-small cell lung cancer, colon cancer, CNS cancer, melanoma, ovarian cancer, renal cancer, prostate cancer and breast cancer). This study provided consistent structure-activity relationships in this series of experimental antitumor compounds.

Nitazoxanide and azithromycin for the early treatment of COVID‐19: A multi‐spectroscopic, biochemical and computational drug–drug interaction study

AbstractHerein, we present an experimental and theoretical drug–drug interaction study between nitazoxanide (NTZ) and azithromycin (AZT) in an aqueous solution. Interaction was studied by using UV/Vis, fluorescence, attenuated total reflectance‐fourier transform infrared (ATR‐FTIR), and circular dichroism (CD) spectroscopy, while molecular docking studies were performed to establish the interaction computationally. A bright yellow color was observed when the two drugs interacted, giving a hyperchromic band at 420 nm. The rate of absorbance was linearly increased by increasing drug concentrations and in a time‐dependent manner. Stability of the interaction complex (i.e., NTZ: AZT) was measured at variable temperatures (25–80°C), pH (5.0–10.0) and ionic strength (0.05–2.0 M NaCl), and not only proved stable but also retained antimicrobial potential with reduced cellular toxicity. Mole ratio and Job's method of continuous variations were used to establish the binding stoichiometry and found to be 2:1. The calculated binding constant (kb = 8,400 M−1) and Gibb's free energy (ΔG° = −22.4 KJ/mol) also suggested an energetically favorable interaction. FTIR spectra of NTZ: AZT complex in comparison with two drugs alone revealed significant interaction which was nicely complemented by molecular docking studies. Interaction was also successfully demonstrated in presence of carrier protein HSA and by spiking the two drugs in real samples of human plasma and urine.

Pan-genome mediated therapeutic target mining in Kingella kingae and inhibition assessment using traditional Chinese medicinal compounds: an informatics approach

Kingella kingae causes bacteremia, endocarditis, osteomyelitis, septic arthritis, meningitis, spondylodiscitis, and lower respiratory tract infections in pediatric patients. Usually it demonstrates disease after inflammation of mouth, lips or infections of the upper respiratory tract. To date, therapeutic targets in this bacterium remain unexplored. We have utilized a battery of bioinformatics tools to mine these targets in this study. Core genes were initially inferred from 55 genomes of K. kingae and 39 therapeutic targets were mined using an in-house pipeline. We selected aroG product (KDPG aldolase) involved in chorismate pathway, for inhibition analysis of this bacterium using lead-like metabolites from traditional Chinese medicinal plants. Pharmacophore generation was done using control ZINC36444158 (1,16-bis[(dihydroxyphosphinyl)oxy]hexadecane), followed by molecular docking of top hits from a library of 36,000 compounds. Top prioritized compounds were ZINC95914016, ZINC33833283 and ZINC95914219. ADME profiling and simulation of compound dosing (100 mg tablet) was done to infer compartmental pharmacokinetics in a population of 300 individuals in fasting state. PkCSM based toxicity analysis revealed the compounds ZINC95914016 and ZINC95914219 as safe and with almost similar bioavailability. However, ZINC95914016 takes less time to reach maximum concentration in the plasma and shows several optimal parameters compared to other leads. In light of obtained data, we recommend this compound for further testing and induction in experimental drug design pipeline. Communicated by Ramaswamy H. Sarma

Lecithin as an Effective Modifier of the Transport Properties of Variously Crosslinked Hydrogels.

Transport properties are one of the most crucial assets of hydrogel samples, influencing their main application potential, i.e., as drug carriers. Depending on the type of drug or the application itself, it is very important to be able to control these transport properties in an appropriate manner. This study seeks to modify these properties by adding amphiphiles, specifically lecithin. Through its self-assembly, lecithin modifies the inner structure of the hydrogel, which affects its properties, especially the transport ones. In the proposed paper, these properties are studied mainly using various probes (organic dyes) to effectively simulate drugs in simple release diffusion experiments controlled by UV-Vis spectrophotometry. Scanning electron microscopy was used to help characterize the diffusion systems. The effects of lecithin and its concentrations, as well as the effects of variously charged model drugs, were discussed. Lecithin decreases the values of the diffusion coefficient independently of the dye used and the type of crosslinking. The ability to influence transport properties is better observed in xerogel samples. The results, complementing previously published conclusions, showed that lecithin can alter a hydrogel's structure and therefore its transport properties.

Comparative Susceptibility of Plasmodium ovale and Plasmodium falciparum Field Isolates to Reference and Lead Candidate Antimalarial Drugs in Ghana.

Malaria treatments resulted in the decline of the deadliest Plasmodium falciparum globally while species, such as P. ovale, infections have been increasingly detected across sub-Saharan Africa. Currently, no experimental drug sensitivity data are available to guide effective treatment and management of P. ovale infections, which is necessary for effective malaria elimination. We conducted a prospective study to evaluate P. ovale epidemiology over 1 year and determined ex vivo susceptibility of the field isolates to existing and lead advanced discovery antimalarial drugs. We report that while P. falciparum dominated both symptomatic and asymptomatic malaria cases, P. ovale in mono or co-infections caused 7.16% of symptomatic malaria. Frontline antimalarials artesunate and lumefantrine inhibited P. ovale as potently as P. falciparum. Chloroquine, which has been withdrawn in Ghana, was also highly inhibitory against both P. ovale and P. falciparum. In addition, P. ovale and P. falciparum displayed high susceptibility to quinine, comparable to levels observed with chloroquine. Pyrimethamine, which is a major drug for disease massive prevention, also showed great inhibition of P. ovale, comparable to effects on P. falciparum. Furthermore, we identified strong inhibition of P. ovale using GNF179, a close analogue of KAF156 imidazolopiperazines, which is a novel class of antimalarial drugs currently in clinical phase II testing. We further demonstrated that the Plasmodium phosphatidylinositol-4-OH kinase (PI4K)-specific inhibitor, KDU691, is highly inhibitory against P. ovale and P. falciparum field isolates. Our data indicated that existing and lead advanced discovery antimalarial drugs are suitable for the treatment of P. ovale infections in Ghana. IMPORTANCE Current malaria control and elimination tools such as drug treatments are not specifically targeting P.ovale. P. ovale can form hypnozoite and cause relapsing malaria. P. ovale is the third most dominant species in Africa and requires radical cure treatment given that it can form liver dormant forms called hypnozoites that escape all safe treatments. The inappropriate treatment of P. ovale would sustain its transmission in Africa where the medical need is the greatest. This is a hurdle for successful malaria control and elimination. Here, we provided experiment data that were lacking to guide P. ovale treatment and disease control policy makers using reference antimalarial drugs. We also provided key experimental data for 2 clinical candidate drugs that can be used for prioritization selection of lead candidate's identification for clinical development.

A Microbial Community Cultured in Gradient Hydrogel for Investigating Gut Microbiome-Drug Interaction and Guiding Therapeutic Decisions.

Despite the recognition that the gut microbiota acts a clinically significant role in cancer chemotherapy, both mechanistic understanding and translational research are still limited. Maximizing drug efficacy requires an in-depth understanding of how the microbiota contributes to therapeutic responses, while microbiota modulation is hindered by the complexity of the human body. To address this issue, a 3D experimental model named engineered microbiota (EM) is reported for bridging microbiota-drug interaction research and therapeutic decision-making. EM can be manipulated in vitro and faithfully recapitulate the human gut microbiota at the genus/species level while allowing co-culture with cells, organoids, and isolated tissues for testing drug responses. Examination of various clinical and experimental drugs by EM reveales that the gut microbiota affects drug efficacy through three pathways: immunological effects, bioaccumulation, and drug metabolism. Guided by discovered mechanisms, custom-tailored strategies are adopted to maximize the therapeutic efficacy of drugs on orthotopic tumor models with patient-derived gut microbiota. These strategies include immune synergy, nanoparticle encapsulation, and host-guest complex formation, respectively. Given the important role of the gut microbiota in influencing drug efficacy, EM will likely become an indispensable tool to guide drug translation and clinical decision-making.

Current and emerging therapies for Achondroplasia: The dawn of precision medicine

Achondroplasia is a rare disease affecting bone growth and is caused by a missense mutation in the fibroblast growth factor receptor 3 (FGFR3) gene. In the past few years, there were multiple experimental drugs entering into clinical trials for treating achondroplasia including vosoritide, the first precision medicine approved for this indication.This perspective presents the mechanism of action, benefit, and potential mechanistic limitation of the drugs currently being evaluated in clinical trials for achondroplasia. This article also discusses the potential impact of those drugs not only in increasing the growth of individuals living with achondroplasia but also in improving their quality of life.

Precocious Puberty.

Precocious puberty is a common presentation to pediatricians with a significant overlap between physiology and pathology. While most girls with precocious puberty have no identifiable cause, boys are more likely to have a pathological cause. The trend of earlier onset of thelarche with slow pubertal tempo has led to a significant increase in the number of girls presenting with precocious puberty. Advanced growth, bone age, uterine maturation, and elevated LH suggest rapidly progressive puberty. The critical issues in evaluating a child presenting with precocious puberty include its confirmation, exclusion of physiological variants, identification of the cause, and determining the need for treatment. Step-wise evaluation with emphasis on clinical parameters provides cost-effective assessment. Gonadotropin-releasing hormone (GnRH) analogs remain the mainstay of treatment for central precocious puberty but should be restricted to individuals with rapidly progressive puberty and compromised final height. The management of rarer forms of peripheral precocious puberty (McCune Albright syndrome, congenital adrenal hyperplasia, and testotoxicosis) involves using experimental drugs under the guidance of specialists.

MF-PCBA: Multifidelity High-Throughput Screening Benchmarks for Drug Discovery and Machine Learning.

High-throughput screening (HTS), as one of the key techniques in drug discovery, is frequently used to identify promising drug candidates in a largely automated and cost-effective way. One of the necessary conditions for successful HTS campaigns is a large and diverse compound library, enabling hundreds of thousands of activity measurements per project. Such collections of data hold great promise for computational and experimental drug discovery efforts, especially when leveraged in combination with modern deep learning techniques, and can potentially lead to improved drug activity predictions and cheaper and more effective experimental design. However, existing collections of machine-learning-ready public datasets do not exploit the multiple data modalities present in real-world HTS projects. Thus, the largest fraction of experimental measurements, corresponding to hundreds of thousands of "noisy" activity values from primary screening, are effectively ignored in the majority of machine learning models of HTS data. To address these limitations, we introduce Multifidelity PubChem BioAssay (MF-PCBA), a curated collection of 60 datasets that includes two data modalities for each dataset, corresponding to primary and confirmatory screening, an aspect that we call multifidelity. Multifidelity data accurately reflect real-world HTS conventions and present a new, challenging task for machine learning: the integration of low- and high-fidelity measurements through molecular representation learning, taking into account the orders-of-magnitude difference in size between the primary and confirmatory screens. Here we detail the steps taken to assemble MF-PCBA in terms of data acquisition from PubChem and the filtering steps required to curate the raw data. We also provide an evaluation of a recent deep-learning-based method for multifidelity integration across the introduced datasets, demonstrating the benefit of leveraging all HTS modalities, and a discussion in terms of the roughness of the molecular activity landscape. In total, MF-PCBA contains over 16.6 million unique molecule-protein interactions. The datasets can be easily assembled by using the source code available at https://github.com/davidbuterez/mf-pcba.

Treatment of pets with the active substance dexpanthenol in wound processes

Animal skin is a complex organ that directly perceives the influence of the external environment and performs a barrier-protective function, helps maintain the balance of the internal environment of the body. Disruption of each of the levels of the protective system leads to the development of inflammatory skin diseases. Treatment of wounds remains one of the most pressing scientific and practical problems of modern veterinary medicine. A number of researchers are searching for and developing new veterinary drugs for the treatment of wounds of various etiologies in small pets, but the use of drugs often causes adverse skin reactions. The purpose of this study was to evaluate the effectiveness of using a veterinary drug (ointment) in the treatment of wounds in domestic animals (dogs, cats, rabbits). The paper uses standard clinical, haematological, and biochemical research methods. The studies used a drug with the active substance dexpanthenol and excipients decamethoxine, Vaseline oil, emulsifier, glycerin, methylparaben, propylparaben, and purified water. The drug under study effectively reduced the wound surface area at all observation periods. It is proved that when using dexpanthenol-based ointment (10%), a complete wound-healing effect was observed in experimental animals on the 14th day of treatment: the area of wound surfaces decreased by 93.1±1.51%. The examined indicators were statistically substantially different from those in the group of untreated animals with skin wounds. The general clinical condition of domestic animals was examined, haematological and biochemical studies of animal blood were conducted under the conditions of applying an experimental drug to damaged skin areas. In animals with skin damage without treatment, the dynamic development of an acute inflammatory process in the body, the development of endogenous intoxication syndrome, signs of wound infection, pronounced exudation and slowing of contraction were observed. The developed veterinary drug with the active substance dexpanthenol simultaneously shows a pronounced antiinflammatory and wound-healing effect and provides a rapid therapeutic effect in wound processes in small pets

Abstract 3412: Patient-derived MicroOrganoSpheres (MOS) enable precision clinical decision-making for multiple myeloma patients

Abstract There are many equipoised multiple myeloma (MM) treatments and nearly all relapse patients undergo cycles of treatment, response, and relapse management. Selecting the right agents and right drug combination is thus of critical importance and an area of unmet need. We currently lack patient-derived MM models that can enable functional precision medicine to help real-time clinical decision-making to guide individual patient treatment. We have created a method to grow MM patient avatars in MicroOrganoSheres (MOS), microscale droplet ECM that sustain the original tumor microenvironment (TME) including both stromal and immune compartments. MOS enable reliable testing of available drug combinations and experimental drugs within 10 days of bone marrow (BM) biopsy, making it feasible to guide treatment decisions in the clinic. In the current study, BMB-derived MM MOS were generated via droplet microfluidics and cultured in vitro, followed by live MOS staining and flow cytometry. Drug screen was performed on MOS with FDA-approved single agents and combinations. Responses of tumorspheres within MOS to drug treatments were quantified by AI-based live imaging algorithms to generate robust drug response curves and derive IC50 and EC50. We successfully established from all 6 MM BM biopsies, MOS conserved CD138/CD38 tumor cell, stromal (e.g., osteoblast) cells, and all major immune cell populations, including T, NK, B, macrophage, dendritic, and myeloid-derived suppressor cells till at least 11 days after the drug assays were completed. The vast number of MOS generated from a single BM biopsy enabled high-throughput drug screening within 10 days. We tested standard-of-care drugs and their combinations applicable to the patient in 9-dose titrations. Live/dead dye fluorescence signal was obtained after 5 days drug dosing and analyzed using Xilis AI analysis pipeline. We generated MOS drug response curve and IC50s from each biopsy. Drug responses from the first 6 MM patient MOS correlates well with their clinical outcome. Furthermore, this MOS platform enabled us to evaluate experimental therapies currently in Phase I-III clinical trials, which revealed responsive patients and insights into optimal combinations that will further improve efficacy. Based on this groundbreaking study, a clinical trial including 40 patients is starting in a month to further validate the predictability of the MM MOS assay for therapeutic decision-making in the clinic. Citation Format: Rui Xi, Xiaobei Wang, Robert Moseley, Renuka Raman, Ray Zhang, Shaima Jaibbar, Shaun Steele, Shengi Ding, Yubing Kang, Xiling Shen. Patient-derived MicroOrganoSpheres (MOS) enable precision clinical decision-making for multiple myeloma patients [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3412.

Abstract 5726: Pan-cancer proteogenomics expands the landscape of therapeutic targets

Abstract Background: Molecularly targeted therapies are critical for improving cancer treatment. Since proteins are the targets of these therapies and functional effectors of genomic aberrations, proteogenomics data from the Clinical Proteomics Tumor Analysis Consortium (CPTAC) provides an unprecedented opportunity to characterize existing and future therapeutic targets for cancer treatment. Approach: CPTAC proteogenomics data from >1000 cancer patients spanning 10 cancer types was used to evaluate current and potential therapeutic targets curated from four databases. Cell line data from DepMap was further integrated to distinguish causations from associations. Computational pipelines were deployed to identify synthetic lethality for targeting tumor suppressor loss and to prioritize tumor associated antigens as immunotherapy targets. Results: We systematically collected 3050 druggable proteins and classified them into 5 tiers to facilitate different applications such as companion diagnostics, drug repurposing, and new therapy development. Many druggable proteins showed poor mRNA-protein correlation, including secreted proteins and proteins whose abundance was correlated with their interaction partners instead of cognate mRNA, highlighting the necessity of direct proteomic quantification of drug targets. 618 druggable proteins showed both overexpression in tumors compared to normal and significant dependency in CRISPR-Cas9 screens of cell lines of the same lineage. Notably, PAK1, a kinase targeted by investigational drugs, demonstrated both overexpression and dependency in all cancer types. A similar analysis of phosphoproteomics data focusing on known activating sites of druggable proteins further revealed targetable dependencies driven by protein hyperactivation. The phosphosite pS50 on PTPN1, a phosphatase targeted by experimental drugs, was increased in 7 cancer types and PTPN1 demonstrated dependency in related cancer cell lines. Based on tumor proteogenomic data and cell line CRISPR-Cas9 screen data, we identified synthetic lethality for difficult to target tumor suppressor losses, revealing TP53 mutations as a candidate biomarker to select breast cancer patients for CHEK1 inhibition, and endometrial cancer patients for treatment with doxorubicin. We identified 140 proteins whose expression was restricted in normal tissues but abnormal in tumors. Experimental analysis of peptides predicted to have high binding affinity to the most common allotype HLA-A02 for 7 prioritized proteins identified 21 peptides from 5 proteins with both strong binding affinity and immunogenicity which could be further investigated as immunotherapy targets. Conclusion: We generate a comprehensive resource of protein and peptide targets that covers multiple therapeutic modalities. This unique resource will pave the way for repurposing of currently available drugs and developing new drugs for cancer treatment. Citation Format: Jonathan T. Lei, Sara R. Savage, Xinpei Yi, Bo Wen, Hongwei Zhao, Lauren K. Somes, Paul W. Shafer, Yongchao Dou, Qiang Gao, Valentina Hoyos, Bing Zhang. Pan-cancer proteogenomics expands the landscape of therapeutic targets. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 5726.

Abstract 5054: The study of synergy mechanism between surufatinib and vinorelbine in the treatment of non-small cell lung cancer

Abstract Background: The incidence and mortality rate of lung cancer ranks first among malignant tumors. Non-small cell lung cancer (NSCLC) is the most common pathological type of lung cancer. Combined use of targeted therapy can enhance the efficacy of chemotherapy drugs while reduce the related side effects in clinical practice. Surufatinib is a potent, small-molecule tyrosine kinase inhibitor (TKI), selectively targeting VEGF receptors (VEGFR) 1, 2, and 3, Fibroblast growth factor receptor 1 (FGFR1), and CSF-1R, which simultaneously prevents tumor angiogenesis and tumor immune evasion. Our clinical study (NCT04922658) has showed the promising effect and safety of surufatinib plus vinorelbine in NSCLC patients, but the potential mechanism is still to be revealed. Herein, tumor-bearing models and cancer cell culture platform would be established for further studying, which may shed light on the synergy mechanism between surufatinib and vinorelbine, and guide the clinical treatment ultimately. Methods: Subcutaneous tumorigenesis in nude mice were successfully established, and then treated with related experimental drugs. The tumor size was observed and CD31 expression of tumor tissue was measured using immunohistochemical analysis. In vitro study, the A549 and PC9 cell multiplication capacities were measured by CCK8 assay and plate cloning assay. The abilities of cancer cell migration and apoptosis were detected using cell scratch assay and flow cytometry kit, respectively. Results: Monotherapy with surufatinib or vinorelbine showed a moderate tumor suppression, whereas the combination therapy of surufatinib and vinorelbine showed the remarkable inhibition of tumor growth in vivo. Surufatinib and the combination therapy obviously decreased the expression of CD31 in tumor issue which might cause the anti-angiogenesis. Monotherapy with surufatinib or vinorelbine significantly inhibited the proliferation of lung cancer cells (A549 cells and PC9 cells) in a concentration-dependent manner, and surufatinib could memorably promote the anti-proliferation activity of vinorelbine. As a result, combination therapy of surufatinib and vinorelbine distinctly inhibited lung cancer cell proliferation when compared with monotherapy with vinorelbine. Consistently, the synergistic effects of surufatinib plus vinorelbine were also observed in cell migration and apoptosis analysis. Conclusions: Synergistic effect of surufatinib and vinorelbine resulted in a conspicuous tumor regression, which is consistent with the clinical observation. In a depth data analysis, surufatinib enhanced the antitumor activity of vinorelbine probably by inhibiting tumor cell proliferation and migration, suppressing tumor angiogenesis and promoting tumor apoptosis in NSCLC patients. Citation Format: Xiaoran Wu, Yanfang Zheng, Lingyu Qin, Ying Li, Zhongjian Yu, Xiongjie Zhu, Rui Cai, Feiyu Niu, Yan Yuan, Peng Jiang, Shaoshi Wen, Xing Lv, Jiancai Zhou. The study of synergy mechanism between surufatinib and vinorelbine in the treatment of non-small cell lung cancer. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 5054.

Abstract 4912: Anticancer activity of Hsp90 inhibitor AUY922 in nasopharyngeal carcinoma and non-small cell lung cancer cells

Abstract Introduction: Nasopharyngeal carcinoma (NPC) and non-small cell lung cancer (NSCLC) are two of the most prevalent cancers in Southern China. Although cisplatin therapy may be initially successful, its effectiveness is often significantly reduced in patients with relapsed disease. There is an urgent need to overcome cisplatin resistance in the treatment of NPC and NSCLC. Luminespib (AUY922) is an experimental drug candidate for the treatment of cancer. It is an inhibitor of heat shock protein 90 (Hsp90), a chaperone protein that plays a role in modifying a variety of proteins involved in tumorigenesis. Hsp90 inhibitors have demonstrated potent activity in preclinical testing and clinical trials against various cancer types. We sought to investigate the preclinical efficacy of AUY922 and evaluate its anticancer effect in cisplatin-resistant NPC and NSCLC cells. Materials and Methods: Two NPC cells (HK1 and Hone1), four NSCLC cells (A549, SW900, NCI-H1975, and HCC4006), one normal lung cells (CCD11Lu), two cisplatin-resistant NPC cells (HK1-R and Hone1-R), and one cisplatin-resistant NSCLC cells (A549-R) were cultured and treated with 100 nM AUY922 for 48 hours. A cell proliferation assay was performed in triplicate to determine the antiproliferative effects of AUY922 on cancer cells. Flow cytometry was used for cell cycle analysis. The apoptotic cells were evaluated by Annexin V and Western blot analysis. Nude mice were used to test the effects of intraperitoneal injection of AUY922 on tumor growth. Results: We found that AUY922 significantly inhibited in vitro growth in all the two NPC (44% and 40%) and four NSCLC (53%, 77%, 50%, and 62%) cells in comparison to the normal lung cells (4%). The two cisplatin-resistant NPC cells (63% and 37%) also showed significant inhibition of viability after treatment with AUY922. However, there was only a slightly reduced viability in the cisplatin-resistant NSCLC cells (14%). Flow cytometry revealed that AUY922 induced G2/M and S phase arrest in HK1 and HK1-R cells. On the other hand, AUY922 also induced pro-apoptotic peaks in A549, A549-R, Hone1, and Hone1-R cells. Interestingly, Annexin V and Western blot analysis showed that cell death was not caused by apoptosis. In vivo, both AUY922 alone and the combination of AUY922 with cisplatin treatments inhibited NPC and NSCLC tumor growth without significant weight loss. In addition, the administration of AUY922 significantly reduced NPC and NSCLC tumor growth in the cisplatin-resistant mouse models. Conclusion: AUY922 exhibits potent anticancer activities in vitro and in vivo. These findings provide evidence that AUY922 could be useful for the treatment of NPC and NSCLC as a single agent or in combination with cisplatin. Most importantly, our results provide a preclinical proof of concept for the efficacy of AUY922 in cisplatin-resistant NPC and NSCLC treatment. Citation Format: William C. Cho, Chi F. Wong. Anticancer activity of Hsp90 inhibitor AUY922 in nasopharyngeal carcinoma and non-small cell lung cancer cells. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4912.

Abstract 390: A comparison of the activity of the quadruplex-targeting experimental drugs QN-302 and CX-5461 (pidnarulex) in wild-type and gemcitabine-resistant pancreatic cancer cell lines

Abstract The compound QN-302, a tetra-substituted naphthalene diimide (ND) derivative has been designed as a potent compound for targeting quadruplex sequences in cancer genes. It has single-digit nM anti-proliferative activity in a panel of human pancreatic cancer (PDAC) cell lines (Ahmed et al., ACS Med Chem Lett, 2020, 11, 1634-1644). It also has significant anti-tumor activity in xenograft, orthotopic and genetic (KPC) models for PDAC. Its mode of action involves the targeting of quadruplex (G4)-forming sites in promoter regions of cancer-associated genes, where their prevalence is over-represented compared to other genes. The structurally unrelated compound CX-5461 has also been reported to be a G4-binding agent, with selective activity in BRCA1/2 deficient cells and tumors (Xu and Hurley, Bioorg Med Chem Lett. 2022; Xu et al., Nat Commun, 2017). It is currently in early-stage clinical trials in breast and other solid cancers with deficiencies in DNA repair (Hilton et al., Nat Commun. 2022). We have previously reported that a precursor to QN-302, the tri-substituted naphthalene diimide compound CM03, is highly active in both wild-type and gemcitabine-resistant PDAC cell lines (Ahmed et al, Sci Rep, 2020). This is explained by the finding using whole-genome transcriptome analysis that the CM03-sensitive genes are largely unaffected in the resistant lines. We now report that the clinical candidate compound QN-302, which is 10-fold more potent than CM03, retains its potency in both MIA-PACA2 and PANC-1 PDAC cell lines with >1000-fold increase in gemcitabine resistance. This is also the case for CX-5461, although this compound is 50-100 times less potent, with GI50 values of 83-95 nM in cell proliferation assays treated for 96 h. Gemcitabine either alone or in combination is still a mainstay of current PDAC treatment. Resistance to gemcitabine is common and is a major contributor to the poor outcomes for most PDAC patients. The ability of QN-302 to retain activity in chemo-resistant PDAC cell lines suggests that it will offer significant advantage in the clinic over gemcitabine-based therapies. QN-302 is bio-available at therapeutic doses and is well tolerated at these levels in animal models. It is being developed for clinical evaluation by Qualigen Therapeutics Inc and is currently at the pre-IND stage. Citation Format: Ahmed Ahmed, Tariq Arshad, Stephen Neidle. A comparison of the activity of the quadruplex-targeting experimental drugs QN-302 and CX-5461 (pidnarulex) in wild-type and gemcitabine-resistant pancreatic cancer cell lines [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 390.