Papers by Subrahmanyam Vangala
Journal of Pharmaceutical and Biomedical Analysis, Oct 1, 2018
We reported first CYP profiling, CYP inhibition study of Guggulsterone. Nineteen phase I & II m... more We reported first CYP profiling, CYP inhibition study of Guggulsterone. Nineteen phase I & II metabolites of guggulsterone were identified after incubation with human liver microsome. Guggulsterone showed high plasma protein binding
Current Pharmaceutical Biotechnology, Apr 20, 2018
Background: Drug transporters function as gatekeepers and modulate drug access into body and vari... more Background: Drug transporters function as gatekeepers and modulate drug access into body and various tissues. Thus, a thorough and precise understanding of transporter liability for compound uptake and efflux is critical during drug development. Methods: In the present study, we assessed the apparent permeability (P app) and compared efflux ratio of various compounds in stably transfected Madin-Darby Canine Kidney (MDCKII) cells overexpressing human P-gp (MDCKII-MDR1), human BCRP (MDCKII-BCRP), wild-type (MDCKII-WT), and Caco-2 cell monolayers. Results: We observed that quinidine, a substrate for MDR1 transporter, showed efflux ratio (P app B-A/ P app A-B) of 838 in MDCKII-MDR1 cells which plummeted to 14 in presence of verapamil, a known inhibitor of MDR1. With MDCKII-WT cells, P app of quinidine dropped from 2 to 1, in the presence of verapamil. Caco-2 cells showed a diminutive decrease in efflux ratio of quinidine from 2.5 to 1.6 by verapamil. Prazosin and dantrolene were evaluated in MDCKII-BCRP cells and were found to have 80-fold higher efflux ratio compared to MDCKII-WT cells. In Caco-2 cells, prazosin and dantrolene showed efflux ratio of 4 and 2, respectively. Rhodamine-123, a fluorogenic probe substrate of MDR1 showed an efflux ratio of 4 in Caco-2 cells and BCRP substrate estrone-3-sulphate showed an efflux ratio of 7. In presence of BCRP inhibitor fumitremorgin-c, the efflux ratio of estrone-3-sulfate dropped to 1 in Caco-2 cells. Conclusion: The very high efflux ratios of MDR1 and BCRP substrates in transfected MDCKII cells clearly demonstrate the potential usefulness of these models to provide more definitive data to evaluate the transporter involvement compared to Caco-2 or MDCKII-WT cells.
Journal of pharmaceutical and biomedical analysis, Jan 25, 2018
The polyphenol E- and Z-gugggulsterone (GS) is an antagonist ligand for the Farnesoid X Receptor ... more The polyphenol E- and Z-gugggulsterone (GS) is an antagonist ligand for the Farnesoid X Receptor (FXR) and known to possess potent hypolipidemic properties as shown in various preclinical and clinical studies. In the present study, we examined drug-like properties of GS by assessing the isomers plasma protein binding, metabolic stability, CYP profiling, CYP inhibition, and phase I and II metabolite identification of GS using liver microsomes and S9 fractions. GS followed Lipinski and Veber rules and were substrates of CYP3A CYP2C19 and CYP2D6 isoforms. GS was also found to be an inhibitor of CYP2C19 with an IC value of 2.1 μM. GS showed high plasma protein binding (<96%), and low to moderate binding with human serum albumin (∼70%). Unbound intrinsic clearances (CL, ) was determined to be low at 0.029 ± 0.0009 and 0.027 ± 0.008 mL/min/mg protein for E- and Z-isomer, respectively in human liver microsomes. Nineteen phase I and II metabolites were identified and hydroxylation was fo...
Current pharmaceutical biotechnology, 2017
Drug transporters function as gatekeepers and modulate drug access into body and various tissues.... more Drug transporters function as gatekeepers and modulate drug access into body and various tissues. Thus, a thorough and precise understanding of transporter liability for compound uptake and efflux is critical during drug development. In the present study, we assessed the apparent permeability (Papp) and compared efflux ratio of various compounds in stably transfected Madin-Darby Canine Kidney (MDCKII) cells overexpressing human P-gp (MDCKII-MDR1), human BCRP (MDCKII-BCRP), wild-type (MDCKII-WT), and Caco-2 cell monolayers. We observed that quinidine, a substrate for MDR1 transporter, showed efflux ratio (Papp B-A/ Papp A-B) of 838 in MDCKII-MDR1 cells which plummeted to 14 in presence of verapamil, a known inhibitor of MDR1. With MDCKII-WT cells, Papp of quinidine dropped from 2 to 1, in the presence of verapamil. Caco-2 cells showed a diminutive decrease in efflux ratio of quinidine from 2.5 to 1.6 by verapamil. Prazosin and dantrolene were evaluated in MDCKII-BCRP cells and were f...
Journal of Analytical & Pharmaceutical Research, 2016
not able to sustain and come up with new drugs? The answer to the above questions primarily lies ... more not able to sustain and come up with new drugs? The answer to the above questions primarily lies in the complexity of biology. Targeted drug discovery has become the mainstream effort of R&D in the last two decades. However, due to redundancy of network pathways and inherent adverse pharmacology of many targets, it is proving difficult for discovering and developing new drug candidates using a single target. About 87% of phase III clinical trials fail due to either lack of efficacy (66%) or safety (21%). Between 2007 and 2010, out of 83 molecules that failed in phase III trials, oncology (28%) and CNS disorders (18%) topped the list of failures (2.3).
Epilepsy Research, 2000
The purpose of this study was to assess the concentration–anti-convulsant effect relationships of... more The purpose of this study was to assess the concentration–anti-convulsant effect relationships of a number of anti-convulsant drugs in the direct cortical stimulation model, to obtain more insight in the properties and predictive value of this model. The time course of the effect of lamotrigine, loreclezole, flunarizine, CGP40116 and CGP39551 was determined after iv. administration in conjunction with their pharmacokinetics.
bioRxiv (Cold Spring Harbor Laboratory), May 15, 2023
Metformin is a widely used and is a safe anti-diabetic drug. It has also been shown to have anti-... more Metformin is a widely used and is a safe anti-diabetic drug. It has also been shown to have anti-inflammatory and anti-viral activities in humans and animal models. Specifically we explored its activity in SARS-CoV-2 initiated COVID19 disease. Here we show that metformin 1. blocks the binding of SARS-CoV-2 spike protein receptor binding domain RBD to human ACE2 receptor 2. We also show that it has anti-inflammatory effects and reduces cytokine secretion as well as blocks the recruitment of monocytes to endothelial cells 3. Finally we show its activity in a hamster in vivo model of SARS-CoV-2 infection as a nasal formulation. Based on the safety and the therapeutic properties relevant to COVID-19 it is feasible to propose a nasal spray of metformin that can be used in treatment of this disease. A nasal spray would deliver the drug to the target organ lung and spare other organs which get exposed upon oral dosing.
bioRxiv (Cold Spring Harbor Laboratory), May 8, 2023
Despite the use of several drugs available to treat type 2 diabetes, many patients are unable to ... more Despite the use of several drugs available to treat type 2 diabetes, many patients are unable to reach their target fasting plasma glucose and HbA1c levels. SGLT1 is the major intestinal transport transmembrane protein which functions in uptake of dietary glucose. If we antagonise the binding of dietary glucose to this transport protein, it is expected that blood glucose lowering will follow. We designed specific inhibitory avian antibodies (IgY) against the extracellular glucose binding domain of SGLT1 and tested their potential in glucose lowering. We demonstrate here the antibodies block uptake of glucose and improve the glycemic profile in vivo and represent a novel approach to inhibiting dietary glucose absorption as treatment for diabetes. .
bioRxiv (Cold Spring Harbor Laboratory), Nov 17, 2022
Non caloric sweeteners (NCS) have been used for decades now as sugar substitutes in foods and bev... more Non caloric sweeteners (NCS) have been used for decades now as sugar substitutes in foods and beverages. The market for such products has grown immensely over time. There are human studies which report their negative effects on glucose metabolism with various results of disturbances in glucose metabolism, weight gain etc. No studies to the best of our knowledge have directly addressed the impact of the NCS on muscle glucose uptake. Muscle tissue can account for over 70 percent of whole-body glucose uptake. Therefore, we examined directly the effect of NCS on muscle cell glucose uptake. We find that aspartame moderately increased insulin stimulated glucose. CC-BY-NC-ND 4.
There are many paths for the transmission of SarsCov2 virus. The main routes are nasal and oral a... more There are many paths for the transmission of SarsCov2 virus. The main routes are nasal and oral and the droplets carrying the virus can also be transmitted thru ocular and skin tissues. The gastrointestinal (small intestine), nasal, ocular and skin tissues all present an acidic pH milieu and therefore any treatment with antibodies thru these routes has to have the antibodies remain active at acid pH as well as be resistant to typical protease digestion.To this end we profiled our anti-SarasCov2 receptor binding domain IgY antibodies for retention of activity at acidic and basic pHs and trypsin digestion. We find that the IgY are strongly resistant to denaturation at acid pH as well as not digested by trypsin. Our data strongly support the use of these IgY in treatment of viral transmission thru the GI, nasal, ocular and skin all tissues where the pH is acidic. We also provide an enabling platform to rapidly asses the suitability of any antibody or protein therapeutic for use at acid...
Type 2 diabetes is currently treated by multiple drugs that are often combined to achieve maximum... more Type 2 diabetes is currently treated by multiple drugs that are often combined to achieve maximum blood glucose lowering in patients. Yet more than 50% of patients are unable to attain target glucose levels. There is clear need for agents which can be added to current drugs to help patients achieve their target blood glucose levels safely.To this end we tested an anti-diabetic nutraceutical for its ability to enhance glucose uptake in muscle cells in combination with currently used cardiometabolic drugs. We show here that the nutraceutical is able to effectively improve glucose uptake of multiple drugs suggesting that it may help in enhancement of glucose lowering by the drugs in patients and achieve optimal glucose levels.
Non caloric sweeteners (NCS) have been used for decades now as sugar substitutes in foods and bev... more Non caloric sweeteners (NCS) have been used for decades now as sugar substitutes in foods and beverages. The market for such products has grown immensely over time. There are human studies which report their negative effects on glucose metabolism with various results of disturbances in glucose metabolism, weight gain etc.No studies to the best of our knowledge have directly addressed the impact of the NCS on muscle glucose uptake. Muscle tissue can account for over 70 percent of whole-body glucose uptake. Therefore, we examined directly the effect of NCS on muscle cell glucose uptake. We find that aspartame moderately increased insulin stimulated glucose uptake by muscle cells in vitro. But sucralose, saccharin and stevia suppressed insulin stimulated glucose uptake.Sucralose is one of the most often used sweetener in foods and beverages globally it is important to understand its effects on glucose metabolism. Therefore, we explored the mechanism of its inhibition of glucose uptake ...
Discovery and development of new drugs is a long, expensive and high risk proposition. Millions o... more Discovery and development of new drugs is a long, expensive and high risk proposition. Millions of dollars spent and decade plus years of time taken to discover a new drug have haunted pharma industry for many years. In part, the reliance on animal models to make go or no go decisions for selecting drugs for human trials has been a problem because animal biology does not capture human disease in entirety. In recognition of this, the last decade has seen the emergence of more human like tools being developed in the hope of better prediction of human outcomes. Towards that end we have developed a 3D bioprinted disease in a dish lung cancer model which uses human cells and includes ability to measure drug efficacy, toxicity and metabolism simultaneously. For drug profiling studies in our disease in a dish model we 3D bioprinted intestinal cells, layered below which were liver cells and finally underneath were target lung cancer cells. The idea was to simulate the path taken by an oral drug which encounters the gut, followed by liver and target organs. We demonstrate here that a 3D bioprinted disease model composed of human derived cells is able to concurrently measure in vitro drug efficacy, toxicity and metabolism. Such humanized models will help make early go or no go decisions on the potential of a drug to enter human trials. .
Alternatives to Laboratory Animals
The fact that animal models fail to replicate human disease faithfully is now being widely accept... more 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. Oncoseek Bio-Acasta Health, which is an innovative biotechnology start-up company based in Hyderabad and Vishakhapatnam, India, organises an annual International Conference on 3Rs Research and Progress. In 2021, this conference was on ‘Advances in Research Animal Models and Cutting-Edge Research in Alternatives’. This annual conference is a platform that brings together eminent scientists and researchers from various parts of the world, to share recent advances from their research in the field of alternatives to animals including new approach methodologies, and to promote practices to help refine a...
Drug repurposing can expedite the process of drug development by identifying known drugs which ar... more Drug repurposing can expedite the process of drug development by identifying known drugs which are effective against SARS-CoV-2. The RBD domain of SARS-CoV-2 Spike protein is a promising drug target due to its pivotal role in viral-host attachment. These specific structural domains can be targeted with small molecules or drug to disrupt the viral attachment to the host proteins. In this study, FDA approved Drugbank database were screened using a virtual screening approach and computational chemistry methods. Five drugs were short listed for further profiling based on docking score and binding energies. Further these selected drugs were tested for their in vitro biological activity. There was significant correlation between the prediction from computational studies and the actual RBD-ACE2 binding inhibition by the drugs. Then, we performed a series of studies that mimic some of the biological events seen in COVID-19 patients such as secretion of IL1β, presentation of a more thromboge...
Glyburide (m/z 494) upon incubation with mouse liver microsomes resulted in hydroxylated metaboli... more Glyburide (m/z 494) upon incubation with mouse liver microsomes resulted in hydroxylated metabolites (m/z 510). All the metabolites were characterized using data dependent EMS-EPI (Enhanced Mass Scan – Enhanced Product Ion) scan mode. Extracted Ion Current from mass spec detector identified the hydroxylated metabolites (m/z 510). LC (Liquid chromatography) with PDA (photo diode array detector) is used as a complimentary technique to confirm and quantify the identified metabolites. Metabolites formed in higher amounts are identified in both UV and MS detector. In the present study PDA detector in UV is used to quantify the hydroxylated metabolites with respect to parent drug glyburide. Chromatogram obtained from mass spec detector is influenced by ionization problems and synthesis of metabolites to quantify them is a lengthy process. In the present study spectrum obtained from PDA detector is demonstrated as an approach to quantify metabolites in the absence of synthetic standards.
Bioanalysis, 2012
The current practice of using calibration curves with narrow concentration ranges during bioanaly... more The current practice of using calibration curves with narrow concentration ranges during bioanalysis of new chemical entities has some limitations and is time consuming. In the present study we describe a split calibration curve approach, where sample dilution and repeat analysis can be avoided without compromising the quality and integrity of the data obtained. A split calibration curve approach is employed to determine the drug concentration in plasma samples with accuracy and precision over a wide dynamic range of approximately 0.6 to 15,000 ng/ml for dapsone and approximately 1 to 25,000 ng/ml for cyclophosphamide and glipizide. A wide dynamic range of concentrations for these three compounds was used in the current study to construct split calibration curves and was successfully validated for sample analysis in a single run. Using this method, repeat analysis of samples can be avoided. This is useful for the bioanalysis of toxicokinetic studies with wide dose ranges and studies...
Since 2019 the world has seen severe onslaught of SARS-CoV-2 viral pandemic. There is an urgent n... more Since 2019 the world has seen severe onslaught of SARS-CoV-2 viral pandemic. There is an urgent need for drugs that can be used to either prevent or treat the potentially fatal disease COVD-19. To this end, we screened FDA approved antiviral drugs which could be repurposed for COVID-19 through molecular docking approach in the various active sites of receptor binding domain (RBD). The RBD domain of SARS-CoV-2 spike protein is a promising drug target due to its pivotal role in viral-host attachment. Specifically, we focussed on identifying antiviral drugs which could a) block the entry of virus into host cells, b) demonstrate anti-inflammatory and/or anti-thrombogenic properties. Drugs which poses both properties could be useful for prevention and treatment of the disease. While we prioritized a few antiviral drugs based on molecular docking, corroboration with in vitro studies including a new 3D human vascular lung model strongly supported the potential of Homoharringtonine, a drug ...
Analytical Biochemistry, 2011
Membranes from a stably transfected cell line that expresses the human organic cation 1 transport... more Membranes from a stably transfected cell line that expresses the human organic cation 1 transporter (hOCT1) have been immobilized on the immobilized artificial membrane (IAM) liquid chromatographic stationary phase to form the hOCT1(+)-IAM stationary phase. Membranes from the parent cell line that does not express the hOCT1 were also immobilized to create the hOCT1(-)-IAM stationary phase. Columns were created using both stationary phases, and frontal displacement chromatography experiments were conducted using [ 3 H]-methyl phenyl pyridinium ([ 3 H]-MPP + ) as the marker ligand and MPP + , verapamil, quinidine, quinine, nicotine, dopamine and vinblastin as the displacers. The K d values calculated from the chromatographic studies correlated with previously reported K i values (r 2 = 0.9987; p < 0.001). The data indicate that the hOCT1(+)-IAM column can be used for the on-line determination of binding affinities to the hOCT1 and that these affinities are comparable to those obtained using cellular uptake studies. In addition, the chromatographic method was able to identify a previously undetected high affinity binding site for MPP + and to determine that hOCT1 bound (R)-verapamil to a greater extent than (S)-verapamil. Published by Elsevier B.V.
Uploads
Papers by Subrahmanyam Vangala