Advances in Animal Models and Cutting-Edge Research in Alternatives: Proceedings of the Second International Conference on 3Rs Research and Progress, Hyderabad, 2021.

The fact that animal models fail to replicate human disease faithfully is now being widely accepted by researchers across the globe. As a result, they are exploring the use of alternatives to animal models. The time has come to refine our experimental practices, reduce the numbers and eventually replace the animals used in research with human-derived and human-relevant 3-D disease models.

A DNAzyme based knockdown model for Fragile-X syndrome in zebrafish reveals a critical window for therapeutic intervention.

Introduction: FXS is the leading cause of intellectual disabilities in males and a major monogenic cause of ASD (Autism spectrum disorders). It occurs due to the loss of FMRP, whose role in early development is not well understood. In this study, we have used a novel DNAzyme based approach to create a larval model of FXS in zebrafish with specific focus on the early developmental window.

Statins exacerbate glucose intolerance and hyperglycemia in a high sucrose fed rodent model.

Statins are first-line therapy drugs for cholesterol lowering. While they are highly effective at lowering cholesterol, they have propensity to induce hyperglycemia in patients. Only limited studies have been reported which studied the impact of statins on (a) whether they can worsen glucose tolerance in a high sucrose fed animal model and (b) if so, what could be the molecular mechanism.

Magnetic nanoparticle formulation for targeted delivery of chemotherapeutic irinotecan to lungs.

Lung cancer is the single largest cause of cancer related deaths in the world. Current treatments include surgery, radiation therapy, chemotherapy using cytotoxic drugs, and monoclonal antibodies. Such treatments have limited efficacy due to diverse nature of lung cells involved and lack of tissue penetration.

Correlation of pharmacokinetics and brain penetration data of adult zebrafish with higher mammals including humans.

Introduction: Adult zebrafish pharmacology is evolving rapidly for creating efficacy and safety models for drug discovery. However, there is very limited research in understanding pharmacokinetics (PK) in adult zebrafish. Methods for understanding PK will help in conducting pharmacokinetic - pharmacodynamic (PK-PD) correlations and improving the quality and applicability of data obtained using zebrafish.

Novel Zebrafish EAE model: A quick in vivo screen for multiple sclerosis.

Introduction: Pre-clinical drug discovery for multiple sclerosis (MS) is a labor intensive activity to perform in rodent models. This is owing to the long duration of disease induction and observation of treatment effects in an experimental autoimmune encephalomyelitis (EAE) model. We propose a novel adult zebrafish based model which offers a quick and simple protocol that can used to screen candidates as a step between in vitro experiments and rodent studies.

Simvastatin may induce insulin resistance through a novel fatty acid mediated cholesterol independent mechanism.

Statins are a class of oral drugs that are widely used for treatment of hypercholesterolemia. Recent clinical data suggest that chronic use of these drugs increases the frequency of new onset diabetes. Studies to define the risks of statin-induced diabetes and its underlying mechanisms are clearly necessary.

Investigational drugs for the management of Huntington's disease: are we there yet?

Introduction: Huntington's disease is a hereditary neurodegenerative disease. It is designated as a rare disease in the US, which means there are < 200,000 patients in the country who suffer from it. The drugs that are currently used to treat this disease were not designed specifically for it but developed for other diseases.

Head and neck cancers, the neglected malignancies: present and future treatment strategies.

Head and neck cancers are the most prevalent cancers in Asia and result in 50% of deaths due to all cancers in that region. However, treatment regimen has not changed in the past two decades and there are no specific drugs for this cancer. As a result of this, treatment outcomes remain poor.

Bioenergetics failure in neurodegenerative diseases: back to the future.

Neurodegenerative disease such as Alzheimer's, Parkinson and Huntington's are all characterized by dysfunctional neurons and loss of cognitive/motor functions. Interestingly these three diseases involve overproduction, aggregation or abnormal degradation of a specific aberrant protein, which participates in disease pathogenesis. The aggregated proteins may induce disease causing pathways such as high oxidative stress and reduced neuronal metabolism.

Bioenergetics breakdown in Alzheimer's disease: targets for new therapies.

Alzheimer's disease is rapidly growing worldwide and yet there is no cure for it. Currently available drugs only provide symptomatic relief and do not intervene in disease process sufficiently enough to prevent or cure it. Characteristic features of this disease are decline in neuronal mass and cognitive functions.

Alzheimer's disease amyloid hypothesis at crossroads: where do we go from here?

Alzheimer's disease has been the focus of several drug discovery approaches by the pharmaceutical industry. Four drug candidates coming out of such efforts have recently failed in late-stage clinical trials for lack of efficacy or safety concerns. These drugs were designed based on the presently dominant scientific hypothesis for Alzheimer's disease called the 'amyloid hypothesis'.

Lipid metabolism and Alzheimer's disease: pathways and possibilities.

Background: Alzheimer's disease (AD) is a growing condition in dire need of new therapeutics for its cure. Currently marketed drugs only provide symptomatic relief and do not intervene in disease-modifying pathways. Deposition of amyloid beta peptides in the brain is considered to be at the core of pathological events in AD.

Lipoic acid synthase (LASY): a novel role in inflammation, mitochondrial function, and insulin resistance.

Objective: Lipoic acid synthase (LASY) is the enzyme that is involved in the endogenous synthesis of lipoic acid, a potent mitochondrial antioxidant. The aim of this study was to study the role of LASY in type 2 diabetes.

Research Design And Methods: We studied expression of LASY in animal models of type 2 diabetes.

Serum levels of interleukin 6, C-reactive protein, vascular cell adhesion molecule 1, and monocyte chemotactic protein 1 in relation to insulin resistance and glucose intolerance--the Chennai Urban Rural Epidemiology Study (CURES).

The aim of this cross-sectional study was to assess the association of insulin resistance (IR) with inflammatory molecules C-reactive protein (CRP), interleukin 6 (IL-6), vascular cell adhesion molecule 1 (VCAM-1), and monocyte chemotactic protein 1 (MCP-1) in urban South Indian subjects. The following groups were selected from the population-based Chennai Urban Rural Epidemiology Study: group 1 composed of 50 healthy subjects with normal glucose tolerance without IR; group 2 consisted of 50 normal glucose-tolerant subjects with IR as defined by homeostasis model assessment of IR (HOMA-IR); group 3 consisted of 50 subjects with impaired glucose tolerance (IGT); and groups 4 and 5 each comprised 50 newly diagnosed and known type 2 diabetic subjects, respectively. The inclusion criteria included nonsmokers; normal resting 12-lead electrocardiogram; and absence of angina, myocardial infarction, or history of any known vascular, infectious, or inflammatory diseases, and not on statins or aspirin.

Novel anti-inflammatory role for glycogen synthase kinase-3beta in the inhibition of tumor necrosis factor-alpha- and interleukin-1beta-induced inflammatory gene expression.

Glycogen synthase kinase-3beta (GSK-3beta) is a serine/threonine kinase with a broad array of cellular targets, such as cytoskeletal proteins and transcription factors. Recent studies with GSK-3beta-null mice showed impaired NFkappaB-mediated survival responses. Because NFkappaB serves a dual role as a key regulator of cytokine-induced inflammatory gene expression and apoptosis, we investigated whether modulation of GSK-3beta expression affects cytokine-induced and NFkappaB-mediated inflammatory gene expression.

A new therapeutic target for atherosclerosis treatment: interview with Uday Saxena. Interviewed by Emma Quigley.

Uday Saxena was appointed Chief Scientific Officer at Dr Reddy's Laboratories in 2000. In this role he provides the leadership and general strategy for the company's drug discovery research into metabolic disorders, cardiovascular disorders, inflammation, cancer and anti-infectives. He is also a member of the company's Senior Management Council.

Discovery of novel phenolic antioxidants as inhibitors of vascular cell adhesion molecule-1 expression for use in chronic inflammatory diseases.

Vascular cell adhesion molecule-1 (VCAM-1) mediates recruitment of leukocytes to endothelial cells and is implicated in many inflammatory conditions. Since part of the signal transduction pathway that regulates the activation of VCAM-1 expression is redox-sensitive, compounds with antioxidant properties may have inhibitory effects on VCAM-1 expression. Novel phenolic compounds have been designed and synthesized starting from probucol (1).

Role of oxidative stress and inflammation in the origin of Type 2 diabetes--a paradigm shift.

In Type 2 diabetes the body either produces too little insulin, or does not respond well to it. Current pharmacological treatments, which are less than optimal, either target defective insulin secretion (sulfonylureas, glinides) or insulin resistance (metformin, thiazolidinediones). Exciting new research is now helping us to understand novel pathways that may contribute to defective insulin secretion as well as decreased response to insulin.

Atherosclerosis -- new targets and therapeutics.

Atherosclerosis is well recognized as an inflammatory disease and circulating markers of inflammation such as C-reactive protein and soluble adhesion molecules are strong predictors of atherosclerotic lesion development and future cardiovascular events. Several cells (endothelial, smooth muscle and macrophages) and proteins (inflammatory cytokines and adhesion molecules) contribute to this inflammatory process and lesion development. Although lipid management with statins does reduce levels of circulating inflammatory markers, this appears to be unrelated LDL-lowering.

Inflammatory cytokines and fatty acids regulate endothelial cell heparanase expression.

Heparan sulfates, the carbohydrate chains of heparan sulfate proteoglycans, play an important role in basement membrane organization and endothelial barrier function. We explored whether endothelial cells secrete a heparan sulfate degrading heparanase under inflammatory conditions and what pathways were responsible for heparanase expression. Heparanase mRNA and protein by Western blot were induced when cultured endothelial cells were treated with cytokines, oxidized low-density lipoprotein (LDL) or fatty acids.

Selective inhibition of endothelial and monocyte redox-sensitive genes by AGI-1067: a novel antioxidant and anti-inflammatory agent.

Atherosclerosis is a disease of oxidative stress and inflammation. AGI-1067 [butanedioic acid, mono[4-[[1-[[3,5-bis(1,1-dimethylethyl)-4-,hydroxyphenyl]thio]-1-methylethyl]thio]-2,6-bis (1,1-dimethylethyl)phenyl] ester] is a metabolically stable derivative of, yet pharmacologically distinct from, the antioxidant drug probucol. It is a member of a novel class of orally active, antioxidant, anti-inflammatory compounds termed vascular protectants and exhibits antiatherosclerotic properties in multiple animal models and in humans.

Lead discovery of alpha,beta-unsaturated sulfones from a combinatorial library as inhibitors of inducible VCAM-1 expression.

alpha,beta-Unsaturated sulfones have been discovered from a combinatorial library as leads for a new series of inhibitors of inducible VCAM-1 expression. Although not essential, further conjugation of the sulfonyl group to another vinyl group or a phenyl group increases the potency dramatically.

AGI-1067: a multifunctional phenolic antioxidant, lipid modulator, anti-inflammatory and antiatherosclerotic agent.

To explore the therapeutic efficacy and potential mechanisms of action of a new class of antiatherosclerotic drugs, AGI-1067 [mono[4-[[1-[[3,5-bis(1,1-dimethylethyl)-4-hydroxyphenyl]thio]-1-methylethyl]thio]-2,6-bis(1,1-dimethylethyl)phenyl] ester] (butanedioc acid) was tested in several animal models of atherosclerosis. AGI-1067, a novel phenolic antioxidant, was well tolerated in a 1-year study in hypercholesterolemic cynomolgus monkeys. It lowered low-density lipoprotein cholesterol (LDLc) by 41 and 90% at oral doses of 50 and 150 mg/kg, respectively and increased high-density lipoprotein cholesterol (HDLc) by 107% at the higher dose.

New approaches for treatment of diabetic nephropathy: the endothelium as a target for drug discovery.

Diabetic nephropathy is the leading cause of end-stage renal disease (ESRD). There are currently no therapeutic Hyperlipidaemiants which directly intervene in the pathogenesis of diabetic nephropathy. The mechanisms behind development of diabetic nephropathy are complex and not completely understood.