Novel Spirocyclic Dimer, SpiD3, Targets Chronic Lymphocytic Leukemia Survival Pathways with Potent Preclinical Effects.

Chronic lymphocytic leukemia (CLL) cell survival and growth is fueled by the induction of B-cell receptor (BCR) signaling within the tumor microenvironment (TME) driving activation of NFκB signaling and the unfolded protein response (UPR). Malignant cells have higher basal levels of UPR posing a unique therapeutic window to combat CLL cell growth using pharmacologic agents that induce accumulation of misfolded proteins. Frontline CLL therapeutics that directly target BCR signaling such as Bruton tyrosine kinase (BTK) inhibitors (e.g., ibrutinib) have enhanced patient survival. However, resistance mechanisms wherein tumor cells bypass BTK inhibition through acquired BTK mutations, and/or activation of alternative survival mechanisms have rendered ibrutinib ineffective, imposing the need for novel therapeutics. We evaluated SpiD3, a novel spirocyclic dimer, in CLL cell lines, patient-derived CLL samples, ibrutinib-resistant CLL cells, and in the Eµ-TCL1 mouse model. Our integrated multi-omics and functional analyses revealed BCR signaling, NFκB signaling, and endoplasmic reticulum stress among the top pathways modulated by SpiD3. This was accompanied by marked upregulation of the UPR and inhibition of global protein synthesis in CLL cell lines and patient-derived CLL cells. In ibrutinib-resistant CLL cells, SpiD3 retained its antileukemic effects, mirrored in reduced activation of key proliferative pathways (e.g., PRAS, ERK, MYC). Translationally, we observed reduced tumor burden in SpiD3-treated Eµ-TCL1 mice. Our findings reveal that SpiD3 exploits critical vulnerabilities in CLL cells including NFκB signaling and the UPR, culminating in profound antitumor properties independent of TME stimuli. SIGNIFICANCE: SpiD3 demonstrates cytotoxicity in CLL partially through inhibition of NFκB signaling independent of tumor-supportive stimuli. By inducing the accumulation of unfolded proteins, SpiD3 activates the UPR and hinders protein synthesis in CLL cells. Overall, SpiD3 exploits critical CLL vulnerabilities (i.e., the NFκB pathway and UPR) highlighting its use in drug-resistant CLL.

Design, synthesis, and evaluation of a mitoxantrone probe (MXP) for biological studies.

Mitoxantrone (MX) is a robust chemotherapeutic with well-characterized applications in treating certain leukemias and advanced breast and prostate cancers. The canonical mechanism of action associated with MX is its ability to intercalate DNA and inhibit topoisomerase II, giving it the designation of a topoisomerase II poison. Years after FDA approval, investigations have unveiled novel protein-binding partners, such as methyl-CpG-binding domain protein (MBD2), PIM1 serine/threonine kinase, RAD52, and others that may contribute to the therapeutic profile of MX. Moreover, recent proteomic studies have revealed MX's ability to modulate protein expression, illuminating the complex cellular interactions of MX. Although mechanistically relevant, the differential expression across the proteome does not address the direct interaction with potential binding partners. Identification and characterization of these MX-binding cellular partners will provide the molecular basis for the alternate mechanisms that influence MX's cytotoxicity. Here, we describe the design and synthesis of a MX-biotin probe (MXP) and negative control (MXP-NC) that can be used to define MX's cellular targets and expand our understanding of the proteome-wide profile for MX. In proof of concept studies, we used MXP to successfully isolate a recently identified protein-binding partner of MX, RAD52, in a cell lysate pulldown with streptavidin beads and western blotting.

Forecasting of AQI (PM2.5) for the three most polluted cities in India during COVID-19 by hybrid Daubechies discrete wavelet decomposition and autoregressive (Db-DWD-ARIMA) model.

Air pollution has emerged as a significant environmental challenge at the global level, and India is majorly affected by it. Numerous emission sources, such as automobiles, industries, fuel-burning for household and commercial activities, and dust due to construction activities, are responsible for air pollution. The lockdown in India which was clamped for controlling the spread of virulent disease also brought down the level of pollutants in air significantly. The proposed approach deals with the application of the hybrid model of Daubechies discrete wavelet decomposition (Db-DWD) and the autoregressive integrated moving average (ARIMA) model for modeling and forecasting the chaotic data of air quality index (PM2.5) from the three most polluted cities (Agra, New Delhi, and Varanasi) in India for pre and within lockdown periods. The estimated outputs of the component series are then reconstructed to obtain the final forecast of the AQI data. The statistical evaluation compares the performance of the simple ARIMA model and the joint Db-DWD-ARIMA model. Also, the coupled model has been applied for forecasting efficacy with Daubechies mother wavelet of orders 5, 8, 10, and 12. The hybrid model reduced forecasting errors and improved accuracy significantly. Secondly, the forecasting efficiencies in this hybrid model have enhanced with the increase in wavelet order. This study will help to assess and take appropriate steps to control air pollution levels and to monitor the growing air pollutants, which will be significant for our existence.

Mycobacterium tuberculosis CitA activity is modulated by cysteine oxidation and pyruvate binding.

As an adaptation for survival during infection, Mycobacterium tuberculosis becomes dormant, reducing its metabolism and growth. Two types of citrate synthases have been identified in Mycobacterium tuberculosis, GltA2 and CitA. Previous work shows that overexpression of CitA, the secondary citrate synthase, stimulates the growth of Mycobacterium tuberculosis under hypoxic conditions without showing accumulation of triacylglycerols and makes mycobacteria more sensitive to antibiotics, suggesting that CitA may play a role as a metabolic switch during infection and may be an interesting TB drug target. To assess the druggability and possible mechanisms of targeting CitA with small-molecule compounds, the CitA crystal structure was solved to 2.1 Å by X-ray crystallography. The solved structure shows that CitA lacks an NADH binding site that would afford allosteric regulation, which is atypical of most citrate synthases. However, a pyruvate molecule is observed within the analogous domain, suggesting pyruvate may instead be the allosteric regulator for CitA. The R149 and R153 residues forming the charged portion of the pyruvate binding pocket were mutated to glutamate and methionine, respectively, to assess the effect of mutations on activity. Protein thermal shift assay shows thermal stabilization of CitA in the presence of pyruvate compared to the two CitA variants designed to decrease pyruvate affinity. Solved crystal structures of both variants show no significant structural changes. However, the catalytic efficiency of the R153M variant increases by 2.6-fold. Additionally, we show that covalent modification of C143 of CitA by Ebselen completely arrests enzyme activity. Similar inhibition is observed using two spirocyclic Michael acceptor containing compounds, which inhibit CitA with ICapp50 values of 6.6 and 10.9 µM. A crystal structure of CitA modified by Ebselen was solved, but significant structural changes were lacking. Considering that covalent modification of C143 inactivates CitA and the proximity of C143 to the pyruvate binding site, this suggests that structural and/or chemical changes in this sub-domain are responsible for regulating CitA enzymatic activity.

Design, synthesis, and evaluation of a mitoxantrone probe (MXP) for biological studies.

Mitoxantrone (MX) is a robust chemotherapeutic with well-characterized applications in treating certain leukemias and advanced breast and prostate cancers. The canonical mechanism of action associated with MX is its ability to intercalate DNA and inhibit topoisomerase II, giving it the designation of a topoisomerase II poison. Years after FDA approval, investigations have unveiled novel protein-binding partners, such as methyl-CpG-binding domain protein (MBD2), PIM1 serine/threonine kinase, RAD52, and others that may contribute to the therapeutic profile of MX. Moreover, recent proteomic studies have revealed MX's ability to modulate protein expression, illuminating the complex cellular interactions of MX. Although mechanistically relevant, the differential expression across the proteome does not address the direct interaction with potential binding partners. Identification and characterization of these MX-binding cellular partners will provide the molecular basis for the alternate mechanisms that influence MX's cytotoxicity. Here, we describe the design and synthesis of a MX-biotin probe (MXP) and negative control (MXP-NC) that can be used to define MX's cellular targets and expand our understanding of the proteome-wide profile for MX. In proof of concept studies, we used MXP to successfully isolate a recently identified protein-binding partner of MX, RAD52, in a cell lysate pulldown with streptavidin beads and western blotting. Highlights: An 8-step synthesis was used to generate a biotinylated-mitoxantrone probe (MXP).A pulldown of MXP demonstrated selectivity for RAD52, but not Replication Protein A.Western blot confirmed the identity of the isolated protein, RAD52.

Cdk5 mediates rotational force-induced brain injury.

Millions of traumatic brain injuries (TBIs) occur annually. TBIs commonly result from falls, traffic accidents, and sports-related injuries, all of which involve rotational acceleration/deceleration of the brain. During these injuries, the brain endures a multitude of primary insults including compression of brain tissue, damaged vasculature, and diffuse axonal injury. All of these deleterious effects can contribute to secondary brain ischemia, cellular death, and neuroinflammation that progress for weeks, months, and lifetime after injury. While the linear effects of head trauma have been extensively modeled, less is known about how rotational injuries mediate neuronal damage following injury. Here, we developed a new model of repetitive rotational head trauma in rodents and demonstrated acute and prolonged pathological, behavioral, and electrophysiological effects of rotational TBI (rTBI). We identify aberrant Cyclin-dependent kinase 5 (Cdk5) activity as a principal mediator of rTBI. We utilized Cdk5-enriched phosphoproteomics to uncover potential downstream mediators of rTBI and show pharmacological inhibition of Cdk5 reduces the cognitive and pathological consequences of injury. These studies contribute meaningfully to our understanding of the mechanisms of rTBI and how they may be effectively treated.

Autoregressive models in environmental forecasting time series: a theoretical and application review.

Though globalization, industrialization, and urbanization have escalated the economic growth of nations, these activities have played foul on the environment. Better understanding of ill effects of these activities on environment and human health and taking appropriate control measures in advance are the need of the hour. Time series analysis can be a great tool in this direction. ARIMA model is the most popular accepted time series model. It has numerous applications in various domains due its high mathematical precision, flexible nature, and greater reliable results. ARIMA and environment are highly correlated. Though there are many research papers on application of ARIMA in various fields including environment, there is no substantial work that reviews the building stages of ARIMA. In this regard, the present work attempts to present three different stages through which ARIMA was evolved. More than 100 papers are reviewed in this study to discuss the application part based on pure ARIMA and its hybrid modeling with special focus in the field of environment/health/air quality. Forecasting in this field can be a great contributor to governments and public at large in taking all the required precautionary steps in advance. After such a massive review of ARIMA and hybrid modeling involving ARIMA in the fields including or excluding environment/health/atmosphere, it can be concluded that the combined models are more robust and have higher ability to capture all the patterns of the series uniformly. Thus, combining several models or using hybrid model has emerged as a routinized custom.

Multi Response Optimization of ECDM Process for Generating Micro Holes in CFRP Composite Using TOPSIS Methodology.

The applications of carbon fiber reinforced polymer composites (CFRPCs) in aerospace, automotive, electronics and lab-on-chip devices require precise machining processes. Over the past decade, there have been numerous attempts to machine CFRPCs using both traditional and unconventional machining techniques. However, because of their limitations, these methods have not gained widespread acceptance. In the present research investigation, Electrochemical Discharge Machining (ECDM) process has been employed to produce micro-holes on CFRPC. The experimental strategy was scheduled using L9 orthogonal array keeping applied voltage, electrolyte concentration and inter-electrode gap as input parameters. The material removal rate (MRR) and overcut were selected as output parameters. The technique for order preference by similarity to the ideal solution (TOPSIS) methodology was executed for multi-response optimization. The overcut and MRR of machined samples improved from 150 µm to 48 µm and 2.232 mg/min to 2.1267 mg/min correspondingly while using the optimum parametric settings of the TOPSIS approach. The shape of drilled micro-holes produced by the TOPSIS process is indicative of a machined surface of superior quality, with a reduction in the number of micro-cracks and a diameter that is uniform.

Parametric Optimisation of Friction-Stir-Spot-Welded Al 6061-T6 Incorporated with Silicon Carbide Using a Hybrid WASPAS-Taguchi Technique.

Friction stir spot welding (FSSW) is one of the most popular fusion joining processes. The process is a solid-state welding process that allows welding of weldable as well as non-weldable materials. As a part of this investigation, weld samples of Al6061-T6 were reinforced with silicon carbide (SiC) powder with an average particle size of 45 µm. Initially, a Taguchi L9 orthogonal array was developed with three factors, i.e., rotational speed of the tool, pre-dwelling time, and diameter of the hole that was filled with SiC before welding. The effects of the SiC particles and process parameters were investigated as tensile-shear load and micro-hardness. The optimisation of parameters in order to maximise the output responses-i.e., strength and hardness of the welded joints-was performed using a hybrid WASPAS-Taguchi method. The optimised process parameters obtained were a 3.5 mm guiding hole diameter, 1700 rpm tool rotation speed, and 14 s of pre-dwelling time.

Health awareness of rural households towards COVID-19 pandemic in India: Evidence from Rural Impact Survey of the World Bank.

This paper aims at analysing the level of awareness of the symptoms and the methods of protection from COVID-19 based on the Rural Impact Survey of the World Bank, collected from 5200 households belonging to six states in India that is, Andhra Pradesh, Bihar, Jharkhand, Madhya Pradesh, Rajasthan, and Uttar Pradesh. Data has been analysed using chi-square test and regression analysis. Results of the analysis indicate that about 70.8% rural households are aware of the symptom of coronavirus, and 81.9% are aware of the preventive measures for controlling the spread of COVID-19. Analysis indicates a significant association between awareness level on symptoms and prevention of COVID-19 and socio-demographics and location. The study further analyses the key determinants of awareness of COVID-19 symptoms and preventive measures using the logistics regression model, indicating that age, gender, education, income, poverty status, access to information, cash relief and medical services are the determining factors of health awareness on COVID-19 pandemic among rural households in India. Considering the importance of self-protecting measures in fighting the pandemic, this paper highlights the importance of strengthening public awareness for containing the spread of the COVID-19 pandemic.

Fbxo7 promotes Cdk6 activity to inhibit PFKP and glycolysis in T cells.

Fbxo7 is associated with cancer and Parkinson's disease. Although Fbxo7 recruits substrates for SCF-type ubiquitin ligases, it also promotes Cdk6 activation in a ligase-independent fashion. We discovered PFKP, the gatekeeper of glycolysis, in a screen for Fbxo7 substrates. PFKP is an essential Cdk6 substrate in some T-ALL cells. We investigated the molecular relationship between Fbxo7, Cdk6, and PFKP, and the effect of Fbxo7 on T cell metabolism, viability, and activation. Fbxo7 promotes Cdk6-independent ubiquitination and Cdk6-dependent phosphorylation of PFKP. Importantly, Fbxo7-deficient cells have reduced Cdk6 activity, and hematopoietic and lymphocytic cells show high expression and significant dependency on Fbxo7. CD4+ T cells with reduced Fbxo7 show increased glycolysis, despite lower cell viability and activation levels. Metabolomic studies of activated CD4+ T cells confirm increased glycolytic flux in Fbxo7-deficient cells, alongside altered nucleotide biosynthesis and arginine metabolism. We show Fbxo7 expression is glucose-responsive at the mRNA and protein level and propose Fbxo7 inhibits PFKP and glycolysis via its activation of Cdk6.

Systemic Administration of a Brain Permeable Cdk5 Inhibitor Alters Neurobehavior.

Cyclin-dependent kinase 5 (Cdk5) is a crucial regulator of neuronal signal transduction. Cdk5 activity is implicated in various neuropsychiatric and neurodegenerative conditions such as stress, anxiety, depression, addiction, Alzheimer's disease, and Parkinson's disease. While constitutive Cdk5 knockout is perinatally lethal, conditional knockout mice display resilience to stress-induction, enhanced cognition, neuroprotection from stroke and head trauma, and ameliorated neurodegeneration. Thus, Cdk5 represents a prime target for treatment in a spectrum of neurological and neuropsychiatric conditions. While intracranial infusions or treatment of acutely dissected brain tissue with compounds that inhibit Cdk5 have allowed the study of kinase function and corroborated conditional knockout findings, potent brain-penetrant systemically deliverable Cdk5 inhibitors are extremely limited, and no Cdk5 inhibitor has been approved to treat any neuropsychiatric or degenerative diseases to date. Here, we screened aminopyrazole-based analogs as potential Cdk5 inhibitors and identified a novel analog, 25-106, as a uniquely brain-penetrant anti-Cdk5 drug. We characterize the pharmacokinetic and dynamic responses of 25-106 in mice and functionally validate the effects of Cdk5 inhibition on open field and tail-suspension behaviors. Altogether, 25-106 represents a promising preclinical Cdk5 inhibitor that can be systemically administered with significant potential as a neurological/neuropsychiatric therapeutic.

Spirocyclic dimer SpiD7 activates the unfolded protein response to selectively inhibit growth and induce apoptosis of cancer cells.

The unfolded protein response (UPR) is an adaptation mechanism activated to resolve transient accumulation of unfolded/misfolded proteins in the endoplasmic reticulum. Failure to resolve the transient accumulation of such proteins results in UPR-mediated programmed cell death. Loss of tumor suppressor gene or oncogene addiction in cancer cells can result in sustained higher basal UPR levels; however, it is not clear if these higher basal UPR levels in cancer cells can be exploited as a therapeutic strategy. We hypothesized that covalent modification of surface-exposed cysteine (SEC) residues could simulate unfolded/misfolded proteins to activate the UPR, and that higher basal UPR levels in cancer cells would provide the necessary therapeutic window. To test this hypothesis, here we synthesized analogs that can covalently modify multiple SEC residues and evaluated them as UPR activators. We identified a spirocyclic dimer, SpiD7, and evaluated its effects on UPR activation signals, that is, XBP1 splicing, phosphorylation of eIF2α, and a decrease in ATF 6 levels, in normal and cancer cells, which were further confirmed by RNA-Seq analyses. We found that SpiD7 selectively induced caspase-mediated apoptosis in cancer cells, whereas normal cells exhibited robust XBP1 splicing, indicating adaptation to stress. Furthermore, SpiD7 inhibited the growth of high-grade serous carcinoma cell lines ~3-15-fold more potently than immortalized fallopian tube epithelial (paired normal control) cells and reduced clonogenic growth of high-grade serous carcinoma cell lines. Our results suggest that induction of the UPR by covalent modification of SEC residues represents a cancer cell vulnerability and can be exploited to discover novel therapeutics.

Small-molecule IKKß activation modulator (IKAM) targets MAP3K1 and inhibits pancreatic tumor growth.

Activation of inhibitor of nuclear factor NF-κB kinase subunit-ß (IKKß), characterized by phosphorylation of activation loop serine residues 177 and 181, has been implicated in the early onset of cancer. On the other hand, tissue-specific IKKß knockout in Kras mutation-driven mouse models stalled the disease in the precancerous stage. In this study, we used cell line models, tumor growth studies, and patient samples to assess the role of IKKß and its activation in cancer. We also conducted a hit-to-lead optimization study that led to the identification of 39-100 as a selective mitogen-activated protein kinase kinase kinase (MAP3K) 1 inhibitor. We show that IKKß is not required for growth of Kras mutant pancreatic cancer (PC) cells but is critical for PC tumor growth in mice. We also observed elevated basal levels of activated IKKß in PC cell lines, PC patient-derived tumors, and liver metastases, implicating it in disease onset and progression. Optimization of an ATP noncompetitive IKKß inhibitor resulted in the identification of 39-100, an orally bioavailable inhibitor with improved potency and pharmacokinetic properties. The compound 39-100 did not inhibit IKKß but inhibited the IKKß kinase MAP3K1 with low-micromolar potency. MAP3K1-mediated IKKß phosphorylation was inhibited by 39-100, thus we termed it IKKß activation modulator (IKAM) 1. In PC models, IKAM-1 reduced activated IKKß levels, inhibited tumor growth, and reduced metastasis. Our findings suggests that MAP3K1-mediated IKKß activation contributes to KRAS mutation-associated PC growth and IKAM-1 is a viable pretherapeutic lead that targets this pathway.

Spinal aspergillosis: a rare complication of COVID-19 infection.

BACKGROUND: Several complications have been reported in COVID-19 infection. Most of the complications include secondary infection. CASE PRESENTATION: We report an 85-year-old male who presented with cauda equina syndrome 7-months after contracting COVID-19 infection. We excised an extradural mass which on examination proved to be Spinal Aspergillosis. CONCLUSIONS: Spinal Aspergillosis should be kept in mind in patients who present with local spinal pain with or without neurological deficit after COVID-19 infection.

Pyrrolo[2,1-f][1,2,4]triazine: a promising fused heterocycle to target kinases in cancer therapy.

Cancer is the second leading cause of death worldwide responsible for about 10 million deaths per year. To date several approaches have been developed to treat this deadly disease including surgery, chemotherapy, radiation therapy, hormonal therapy, targeted therapy, and synthetic lethality. The targeted therapy refers to targeting only specific proteins or enzymes that are dysregulated in cancer rather than killing all rapidly dividing cells, has gained much attention in the recent past. Kinase inhibition is one of the most successful approaches in targeted therapy. As of 30 March 2021, FDA has approved 65 small molecule protein kinase inhibitors and most of them are for cancer therapy. Interestingly, several kinase inhibitors contain one or more fused heterocycles as part of their structures. Pyrrolo[2,1-f][1,2,4]triazine is one the most interesting fused heterocycle that is an integral part of several kinase inhibitors and nucleoside drugs viz. avapritinib and remdesivir. This review articles focus on the recent advances made in the development of kinase inhibitors containing pyrrolo[2,1-f][1,2,4]triazine scaffold.

Inhibitors, PROTACs and Molecular Glues as Diverse Therapeutic Modalities to Target Cyclin-Dependent Kinase.

The cyclin-dependent kinase (CDK) family of proteins play prominent roles in transcription, mRNA processing, and cell cycle regulation, making them attractive cancer targets. Palbociclib was the first FDA-approved CDK inhibitor that non-selectively targets the ATP binding sites of CDK4 and CDK6. In this review, we will briefly inventory CDK inhibitors that are either part of over 30 active clinical trials or recruiting patients. The lack of selectivity among CDKs and dose-limiting toxicities are major challenges associated with the development of CDK inhibitors. Proteolysis Targeting Chimeras (PROTACs) and Molecular Glues have emerged as alternative therapeutic modalities to target proteins. PROTACs and Molecular glues utilize the cellular protein degradation machinery to destroy the target protein. PROTACs are heterobifunctional molecules that form a ternary complex with the target protein and E3-ligase by making two distinct small molecule-protein interactions. On the other hand, Molecular glues function by converting the target protein into a "neo-substrate" for an E3 ligase. Unlike small molecule inhibitors, preclinical studies with CDK targeted PROTACs have exhibited improved CDK selectivity. Moreover, the efficacy of PROTACs and molecular glues are not tied to the dose of these molecular entities but to the formation of the ternary complex. Here, we provide an overview of PROTACs and molecular glues that modulate CDK function as emerging therapeutic modalities.

Prediction of COVID-19 pervasiveness in six major affected states of India and two-stage variation with temperature.

Coronavirus disease knocked in Wuhan city of China in December 2019 which spread quickly across the world and infected millions of people within a short span of time. COVID-19 is a fast-spreading contagious disease which is caused by SARS-CoV-2 (severe acute respiratory syndrome-coronavirus-2). Accurate time series forecasting modeling is the need of the hour to monitor and control the universality of COVID-19 effectively, which will help to take preventive measures to break the ongoing chain of infection. India is the second highly populated country in the world and in summer the temperature rises up to 50°, nowadays in many states have more than 40° temperatures. The present study deals with the development of the autoregressive integrated moving average (ARIMA) model to predict the trend of the number of COVID-19 infected people in most affected states of India and the effect of a rise in temperature on COVID-19 cases. Cumulative data of COVID-19 confirmed cases are taken for study which consists of 77 sample points ranging from 1st March 2020 to 16th May 2020 from six states of India namely Delhi (Capital of India), Madya Pradesh, Maharashtra, Punjab, Rajasthan, and Uttar Pradesh. The developed ARIMA model is further used to make 1-month ahead out of sample predictions for COVID-19. The performance of ARIMA models is estimated by comparing measures of errors for these six states which will help in understanding future trends of COVID-19 outbreak. Temperature rise shows slightly negatively correlated with the rise in daily cases. This study is noble to analyse the variation of COVID-19 cases with respect to temperature and make aware of the state governments and take precautionary measures to flatten the growth curve of confirmed cases of COVID-19 infections in other states of India, nearby countries as well.

Structure activity relationship (SAR) study identifies a quinoxaline urea analog that modulates IKKß phosphorylation for pancreatic cancer therapy.

Genetic models validated Inhibitor of nuclear factor (NF) kappa B kinase beta (IKKß) as a therapeutic target for KRAS mutation associated pancreatic cancer. Phosphorylation of the activation loop serine residues (S177, S181) in IKKß is a key event that drives tumor necrosis factor (TNF) α induced NF-κB mediated gene expression. Here we conducted structure activity relationship (SAR) study to improve potency and oral bioavailability of a quinoxaline analog 13-197 that was previously reported as a NFκB inhibitor for pancreatic cancer therapy. The SAR led to the identification of a novel quinoxaline urea analog 84 that reduced the levels of p-IKKß in dose- and time-dependent studies. When compared to 13-197, analog 84 was ∼2.5-fold more potent in TNFα-induced NFκB inhibition and ∼4-fold more potent in inhibiting pancreatic cancer cell growth. Analog 84 exhibited ∼4.3-fold greater exposure (AUC0-∞) resulting in ∼5.7-fold increase in oral bioavailability (%F) when compared to 13-197. Importantly, oral administration of 84 by itself and in combination of gemcitabine reduced p-IKKß levels and inhibited pancreatic tumor growth in a xenograft model.