Day 1 :
Keynote Forum
Shyam B Mehta
Teva Pharmaceuticals, USA
Keynote: A case study on manufacturing issues during processing of a lyophilized drug product
Time : 10:10-10:50
Biography:
Shyam B Mehta has completed his PhD in Pharmaceutical Sciences with expertise in areas specifi c to Biologic drugs. His work focuses on drug product development, fill-finish manufacturing of drug product, formulation development including protein stability and biophysical characterization of protein molecules
Abstract:
Purpose: Despite the popularity and advent of liquid formulations for biological therapeutic products, lyophilized presentations continue to be a robust alternative, especially for products in early stages of development or with in-adequate liquid stability. In this context, there continues to be on-going interest in the pharmaceutical industry about understanding potential failure modes and mitigations for lyophilized products. Here we present a real-life case study of observations of product on stopper for a lyophilized product and provide further analysis if this constitutes only a cosmetic/pharmaceutical elegance issue or can potentially be product quality impacting. Th e primary failure modes that were the focus of this investigation were potentially improper protein recovery during reconstitution and/or sterility concerns due to container closure integrity compromise. In this context, we also discuss potential mitigations. Methods: Filling of product in vials was carried out using a peristaltic pump fi lling unit and subsequently the product was lyophilized. Post lyophilization, upon visual inspection, around 40% of vials had cosmetic defect with residual product around stopper of the vial and the remaining 60% vials with no cosmetic defect were called acceptable vials. Within the product group that had product on stopper, there were diff erent degrees of this event observed. Hence for this evaluation, the lyophilized vials with the worst-case product on stopper profi les were evaluated along with acceptable vials. Testing performed were protein concentration (UV absorbance at 280), residual moisture (Karl Fischer), sterility (membrane fi ltration) and container closure integrity (using blue dye ingress). Results & Conclusion: Product quality attributes such as sterility, residual moisture, protein concentration was also similar between the two groups of vials. However, container closure integrity of the product on stopper vials was compromised due to the presence of product around stopper of the vial. Th e results of this study provide two important fi ndings: Defects such as product on stopper under certain worst-case conditions may pose product quality concerns and not just cosmetic quality concerns, and Container closure integrity test methods may be more sensitive in such cases compared to sterility, which also explains the regulatory agencies preference for this method for assessing sterility assurance. As an outcome of this investigation, the following corrective actions were implemented for this case: use of larger sized vial that can potentially mitigate the product handling issues.
Keynote Forum
Mallikarjun Narayan Dixit
Accutest Biologics Private Limited, India
Keynote: Challenges in development of biologics and biosimilars: An overview stability and validation
Time : 10:50-11:30
Biography:
Dixit (graduate from University of Illinois) is a scholastic personality with several patents and publications to his credit and with more than 25 years of rich experience in pharmaceuticals, biopharmaceutical and CRO industries in the area of drug discovery and bio-analytical services. He has extensively worked on method development and validation of various assay platforms such as LC-MS, ELISA, MSD, SPR, RIA/RIPA and Cell based assays for Immunogenicity and pharmacokinetics evaluation of non-clinical and clinical study samples under GLP and GCLP compliant practices for regulatory submission studies. As a test facility management and head of bioanalytical laboratory he has successfully faced multiple sponsor’s and regulatory audits. Dr. Dixit with his expertise is currently guiding the team biologics in the delivery of quality compliant bioanalytical and Characterization services for biologics and biosimilars for submission studies
Abstract:
The global prescription drug market is expected to grow by 6% to reach nearly USD 1.05 trillion by 2022. Th e top 20 drugs are manufactured by 14 companies and account for a total 10% of global prescription drug market in 2016. Th e total revenue generated by top 20 products was estimated to be USD 0.128 trillion. Biologic drugs are cost eff ective opportunity for patients in the area of oncology, diabetes, infl ammatory disorders, Autoimmune diseases, cardiovascular diseases and represent a total market value of more than $50 billion. Th ere are 150 marketed biologic products worldwide, with almost 500 products under development. About 21 important biologics will loose patent protection by the year 2019. Th e biopharmaceuticals show much greater effi cacy and effi ciency when compared to conventional pharmaceuticals. Th e use of biopharmaceuticals is continuously increasing and has resulted in a huge market demand. Biological drug products including Biosimilars, are larger in size and more complex than conventional small molecule drugs and the majority of these originate from living organisms. During the product development phase, some of the process changes are inevitable such as Cell line change, Critical Raw Material change, Primary packaging components change, Scale up/ down, Process parameter change, Formulation, Presentation, Device and Form Change Site Change or Critical Equipment change. Multiple complex manufacturing processes involved in biologics drug product development may signifi cantly aff ect the product quality attributes, product development process and the product safety and/or effi cacy. Th ese may also aff ect the other quality attributes such as physical, chemical, biological, or microbiological property of the biologic drug product. Th ere are multiple challenges at every stage of development of high quality biologics drug products. Th ese include the initial clone development, clone expression, product development, processing, purifi cation, characterization, post translation modifi cations, sensitive bioanalytical methods and stringent regulatory expectations. Similarly, there are several challenges in the clinical development phase also. Th ese include Study Indication, Study Design, reference product, regulatory approvals, patient recruitment, Inclusion and Exclusion Criteria,
Statistical consideration, Immunogenicity, Safety assessment and Post marketing study requirement. Th e advancement in the technology and availability of scientifi c approaches have helped to a great extent in eff ectively combating the challenges andkeep the biologics drug development process at a rapid pace. Th e presentation on challenges in developing Biologics and Biosimilars gives an overview of scope for Biologics and Biosimilars drugs, various challenges and methods to overcome many of these challenges.
- Biopharmaceutics | Drug Discovery & Development | Biologic Drugs | Biological Medicine | Array of Clinical Trials in Biopharmaceutics
Location: Hilton Philadelphia Airport
Session Introduction
Mallikarjun Narayan Dixit
Accutest Biologics Private Limited, India
Title: Immunogenicity of protein biotherapeutics: Case studies on ADA methods
Time : 11:50-12:20
Biography:
Dixit (graduate from University of Illinois) is a scholastic personality with several patents and publications to his credit and with more than 25 years of rich experience in pharmaceuticals, biopharmaceutical and CRO industries in the area of drug discovery and bio-analytical services. He has extensively worked on method development and validation of various assay platforms such as LC-MS, ELISA, MSD, SPR, RIA/RIPA and Cell based assays for Immunogenicity and pharmacokinetics evaluation of non-clinical and clinical study samples under GLP and GCLP compliant practices for regulatory submission studies. As a test facility management and head of bioanalytical laboratory he has successfully faced multiple sponsor’s and regulatory audits. Dr. Dixit with his expertise is currently guiding the team biologics in the delivery of quality compliant bioanalytical and Characterization services for biologics and biosimilars for submission studies
Abstract:
With the advent of Biotechnology more and more human recombinant protein biotherapeutics have replaced the animal derived drug products. Th is has helped in global availability of quality drug products at a competitive price. Th e advantages of the recombinant protein biotherapeutics are high purity, high specifi c activity, steady supply and batch-to-batch consistency. All protein biotherapeutics are potentially immunogenic and hence could elicit immunogenicity response. Th ese responses could be of diff erent types such as binding antibodies or neutralizing antibodies. Immunogenicity of protein biotherapeutics is a major concern especially when the biological function of the drug and the endogenous counterpart are neutralized by antidrug antibodies (ADA). Hence the regulatory agencies insist that the immunogenicity response should be assessed by validated sensitive assay formats during the diff erent stages of drug development and the antibody response be characterized. An effort has been made in this presentation to discuss on immunogenicity to protein biotherapeutics, regulatory concerns, strategy to evaluate the immunogenicity, overview of methods available, factors infl uencing immunogenicity and to share the experience of developing an ADA method using ELISA and GYROS assay format to assess immunogenicity of two drug products
Nayanabhirama Udupa
Manipal University, India
Title: Effective oral drug delivery for the treatment of diabetes mellitus
Time : 12:20-12:50
Biography:
Nayanabhirama Udupa was at Manipal University as Professor and Head of the institution, Manipal College of Pharmaceutical Sciences for couple of tenures since his joining as Professor from 1987. He has published more than 510 papers peer reviewed journals, presented 411 papers in different conferences throughout the world, contributed 12 books in pharmacy fi eld and gave more than 125 lectures. He has received more than 57 research grants having 9 patents. He has guided 36 PhD candidates and 80 M pharm students for the completion of their thesis.
Abstract:
Diabetes mellitus is a metabolic disorder, is becoming a serious threat to mankind health. Among all types of diabetes, type
2 diabetes is main complication which can be controlled with diet, exercise and oral medications. Oral medications are not insulin pills, rather four classes of drugs designed to improve the body’s utilization of what insulin is still present. These are sulfonylurea, metformin, troglitazone and acarbose but these oral medications will not be eff ective when islets of pancreas stop
making suffi cient insulin. So we have come with a dietary supplement which can be used as a soup for management of diabetes.
Th e composition of the supplement includes all dietary ingredients such as wheat fl our commonly known as bread wheat, barley powder, guar gum it’s a polysaccharide of galactose and mannose, which is water soluble polymer, exhibits a viscosifying
eff ect in water, onion powder commonly known as bulb onion, which contains quercetin, allicin, allinin and salt as required and turmeric powder that are harmless when given to patients. Our preliminary in vitro studies suggested good activity in modifying glucose absorption from the gut and we also carried out a prospective observational pilot study, to study the eff ect of dietary supplement on glycemic control in pre-diabetes (PD) and mildly uncontrolled diabetics (MUCD) and the results have shown that the dietary supplementation could be an adjuvant therapy which is safe, healthy and economic alternative for DM patients to control blood glucose levels and also that can minimize even the dose of OHAs.
Santosh K Singh
Banaras Hindu University, India
Title: Evaluation of anticancer and apoptosis inducing activity of Amoora rohituka W.A. in human breast cancer cell line, MCF-7
Time : 13:50-14:20
Biography:
Santosh K Singh obtained his Master’s in Botany from Veer Bahadur Singh Purvanchal University, Jaunpur in India in 1992. He was awarded a PhD degree in Botany from Banaras Hindu University, Varanasi, India in 1998. He is currently appointed as Senior Research Offi cer in Centre of Experimental Medicine and Surgery, IMS, BHU and leading the center which have facilities for cell culture, natural product research, molecular biology research, plant endophytes research and animal house also for animal study works. He is working on traditionally used herbal medicines, bio-fabrication of highly structured nanomaterials and their evaluation for anti-cancer activity.
Abstract:
Cancer is one of the second most leading causes of death in India and around the world aft er cardiovascular disease while breast cancer is the most common cancer in women worldwide, with nearly 1.67 million new cases diagnosed in 2012. It was projected that the burden of breast cancer will increase by 50% in 2030. Chemotherapy continues to be the most common pharmacological approach for the treatment and most of the chemotherapeutic drugs for cancer treatment are molecules isolated or derived from plants. More new drugs are needed for the treatment because of the development of chemo-resistance. Natural products have continued to receive an increasing attention for their potential as novel cancer preventive and therapeutic agents which show the importance of traditionally used medicinal plants as the prime sources for development of anticancer drugs. Amoora rohituka WA is a well-known medicinal plant which possesses numerous therapeutic actions including antiinflammatory, sedative, hypnotic and narcotic. It was evaluated for anticancer activity in this study. Th e hydro-alcoholic (1:1) crude extract of the plant bark (100 g) was fractionated using column chromatography using benzene, ethyl acetate and methanol solvents respectively. The fractions were dried under vacuum in rotary evaporator and evaluated for their anticancer activity using MTT assay against human breast cancer cell line, MCF-7. Mode of cell death was evaluated by ethidium bromide and acridine orange double-staining. Th is study show that ethyl acetate fraction of bark extract (RBEA) induces cytotoxicity in a dose-dependent manner in cancer cell lines. Ethidium bromide and acridine orange double-staining demonstrated that RBEA potentiate apoptosis rather than necrosis in cancer cells.
Yesenia Elizabeth Cevallos Villacrés
University of Calabria, Italy
Title: A protocol stack paradigm of gene expression
Time : 14:20-14:50
Biography:
Yesenia Elizabeth Cevallos Villacrés is an Electronic and Telecommunication Engineer from National Polytechnic in Ecuador, 2001. She has an Information Technology Master’s degree from Technical University of Ambato in Ecuador, 2010. She is an Associate Professor at National University of Chimborazo, Ecuador. Currently, she is a PhD student at University of Calabria, Italy. She has articles published in network and digital communications area in journals in Ecuador. Her main publications as PhD student is: A digital communication analysis of gene expression of proteins in biological systems: A layered network model view.
Abstract:
In this study, we propose the design of a protocol stack network model to explain gene expression. Gene expression is the process by which information carried by deoxyribonucleic acid (DNA) is transformed into proteins. Proteins are produced in the cell and excreted to perform a biological function. Th e nucleus of eukaryotic cells is the source of biological information, which must be modifi ed via intracellular communication to reach an adequate cellular or extracellular destination. This process implies that information is transferred through a biological path, and aft er it is received, a biological function is performed.
The transmission of proteins (specifi cally peptide hormones) to a target organ through blood stream involves the golgi apparatus, which is similar to the digital communication process wherein a transmitter in a network sends information to a destination device in another network through a router. Th e modeling that we have proposed is supported by very well know theories in communications as Shannon’s theorem, and network characteristics (e.g., independent functions of layers in a stack, addressing, fl ow control, error control, and traffi c control). Our analysis is focused in the importance of the addressing in networks applied to biological systems to reduce the incidence of side eff ects of drugs used to treat disease. Th e objective is to improve the quality of treatment for patients from health and socio-economic perspectives.
Shyam B Mehta
Teva Pharmaceuticals, USA
Title: Product on stopper’ in a lyophilized biologic drug product: Cosmetic defect or a product quality concern?
Time : 14:50-15:20
Biography:
Shyam B Mehta is a PhD student in Pharmaceutical Sciences with expertise in areas specifi c to biologic drugs. His work focuses on drug product development, fi ll-fi nish manufacturing of drug product, formulation development including protein stability and biophysical characterization of protein molecules
Abstract:
Despite the popularity and advent of liquid formulations for biological therapeutic products, lyophilized presentations
continue to be a robust alternative, especially for products in early stages of development or with in-adequate liquid stability. In this context, there continues to be on-going interest in the pharmaceutical industry about understanding potential failure modes and mitigations for lyophilized products. Here, we present a real life case study of observations of “product on stopper” for a lyophilized product and provide further analysis if this constitutes only a cosmetic/ pharmaceutical elegance issue or can potentially be product quality impacting. Th e primary failure modes that were the focus of this investigation were potentially improper protein recovery during reconstitution and/or sterility concerns due to container closure integrity compromise. In this context, we also discuss potential mitigations. Methods: Filling of product in vials was carried out using a peristaltic pump fi lling unit and subsequently the product was lyophilized. Post lyophilization, upon visual inspection, around 40% of vials had cosmetic defect with residual product around stopper of the vial and the remaining 60% vials with no cosmetic defect were called ‘acceptable vials’. Within the product group that had product on stopper, there were diff erent degrees of this event observed. Hence for this evaluation, the lyophilized vials with the worst case product on stopper profi les were evaluated along with “acceptable vials”. Testing performed were protein concentration (UV absorbance at 280), residual moisture (Karl Fischer), sterility (membrane fi ltration) and container closure integrity (using blue dye ingress). Results & Conclusion: Product quality attributes such as sterility, residual moisture, protein concentration were also similar
between the two groups of vials. However, container closure integrity of the ‘product on stopper’ vials was compromised due to the presence of product around stopper of the vial. Th e results of this study provides two important fi ndings- 1) Defects such as “product on stopper” under certain worst case conditions may pose product quality concerns and not just cosmetic quality concerns, and 2) Container closure integrity test methods may be more sensitive in such cases compared to sterility, which also explains the regulatory agencies preference for this method for assessing sterility assurance. As an outcome of this investigation, the following corrective actions were implemented for this case: use of larger sized vial that can potentially mitigate the product handling issues.
Hong Ngoc Thuy Pham
Nha Trang University, Australia
Title: Maximizing saponin yield and antioxidant capacity of Catharanthus roseus (L.) G. Don (Patricia White cultivar) stem by ultrasound-assisted extraction
Time : 15:20-15:50
Biography:
Hong Ngoc Thuy Pham has graduated with a BSc in Food Technology from Nha Trang University, Vietnam in 2004 and then completed her Master’s degree in Postharvest Technology from Nha Trang University, Vietnam in 2009. She is currently a Lecturer of Nha Trang University, Vietnam and a PhD student at the University of Newcastle, Australia. She has published 10 papers in domestic and international journals. She is now working on the research project: Extraction of anticancer compounds from selected medicinal plants as novel agents against pancreatic cancer cells at the University of Newcastle, Australia.
Abstract:
Catharanthus roseus (L.) G Don (C. roseus) is traditionally known as a medicinal plant; however only its leaf has been investigated for its bioactive compounds. Other plant components, such as the stem and the root have not been studied. Our previous study conducted on the stem revealed a high level of saponins, which have been linked with various biological activities such as antimicrobial, ant herbivore and/or cytotoxic activities. Ultrasound-assisted extraction (UAE) is known as an advanced extraction method because of its eff ectiveness, simplicity and up-scale capability, thus UAE shows potential for the eff ective extraction of saponins from the stem of C. roseus. Response surface methodology (RSM) has been commonly applied to optimize extraction conditions as it can show the interactive eff ects between the tested parameters. Th is study aimed to optimize UAE conditions including temperature, time and power in combination with ratio of sample-to-solvent for the effi cient extraction of saponins from stem of C. roseus using RSM with Box-Behnken design. Th e results indicated that either ultrasonic temperature or sample-to-solvent ratio had a signifi cant infl uence on saponin yield, DPPH and ABTS radical scavenging activities (p<0.05). With the exception of ultrasonic power, the other three parameters (UAE temperature, sampleto- solvent ratio and time) statistically aff ected ferric reducing antioxidant power (FRAP) and cupric ion reducing antioxidant capacity (CUPRAC). A maximized saponin yield (138.21 mg ESE/g) and high levels of antioxidant activity (8.34, 12.06, 10.53 and 19.15 mg TE/g ABTS, DPPH scavenging capacity, FRAP and CUPRAC respectively) could be obtained under the optimal extraction conditions (ultrasonic temperature of 55°C, ultrasonic time of 35 min, sample-to-solvent ratio of 1/100 (g/mL) and ultrasonic power of 150W). Th e saponin-enriched extract from C. roseus stem can be potentially used for further investigation and application in the nutraceutical and pharmaceutical industries.
Rubi Mahato
Fairleigh Dickinson University, USA
Title: Nanoemulsion as targeted drug delivery system for cancer therapeutics
Time : 16:10-16:40
Biography:
Rubi Mahato received her interdisciplinary Ph.D. in Pharmaceutical Sciences and Chemistry from University of Missouri-Kansas City in 2014. After that, she joined as an assistant professor at the school of Pharmacy at Fairleigh Dickinson University. She has authored several research and review articles and book chapters. She also serves as a regular reviewer for many prestigious journals, as well as editorial board member for various journals. Her research interests include drug release analysis from transdermal patch, development of targeted delivery systems, ayurvedic treatment for arthritis, and pedagogical research
Abstract:
Nanoemulsion serves as an attractive vehicle for the delivery of drugs, nucleic acids as well as imaging agents. Recently nanoemulsions have been extensively used for cancer diagnostics, imaging and therapy, especially due to their favorable properties to efficiently solubilize poorly aqueous soluble drugs, biocompatibility, high stability in vitro and in vivo, and their ability to accumulate in pathological areas with defective vasculatures. Since nanoemulsions are submicron emulsions with the droplet size falling in colloidal dispersion range, they impart the benefit of overcoming the anatomical and physiological barriers associated with drug delivery to the complex diseases such as cancer. Moreover, nanoemulsions can be engineered to carryout multiple functions by surface modification and encapsulation of pharmaceutical ingredients. Surface modification can be done by imparting the surface charge, attaching a targeting ligand, cell penetrating moieties, stimuli-sensitive groups and fluorescent dye, whereas the core can be loaded with drug, contrast agent and imaging agents. Such multifunctionality of nanoemulsion can be tailored to fit the requirement, hence smart nanoemulsions can be prepared.
Kamna Srivastava
University of Delhi, India
Title: Role of angiotensinogen gene polymorphism in blood pressure lowering response to angiotensin converting enzyme inhibitors
Time : 16:40-17:10
Biography:
Kamna Srivastava has completed her B Pharm, M Pharm and PhD from Department of Pharmaceutics, Institute of Technology, Banaras Hindu University, India. She held her Post-doctoral positions in National Institute of Immunology, All India Institute of Medical Sciences, New Delhi. Presently, she is working as an Assistant Professor in Molecular Cardiology Lab in Dr. B R Ambedkar Centre for Biomedical Research, Delhi University. Her on-going project is focused on the identifying the potential biomarkers for cardiovascular diseases. She has more than 30 research publications to her credit and recipient of grants from DST, CSIR and ICMR India.
Abstract:
Background: Hypertension is one of the major risk factors for cardiovascular diseases. It has been suggested that genetic backgrounds, which have an association with essential hypertension, may also determine the responsiveness to ACE inhibitor. We determined the association of angiotensinogen gene polymorphism with essential hypertension and the relationship between polymorphism in the angiotensinogen (M235T) gene and blood pressure response to ACE inhibitor (enalapril) in patients with essential hypertension. Th e present study is also about a particular genetic polymorphism (A1166C), gene expression and protein expression of the angiotensin II type I receptor (AT1R) and its association with essential hypertension in a Northern Indian population. Methods: 250 patients with essential hypertension and 250 normal healthy controls from Delhi and surrounding areas were recruited for the investigation. Blood pressure was recorded before and aft er 6 weeks of treatment with ACE inhibitors, Enalapril. Genotyping were carried out by PCR and RFLP technique and expression analysis at mRNA levels by real-time PCR. Results and Conclusions: Statistically signifi cant association of T allele was observed with essential hypertension [x2=14.67, p=0.00013, Odds ratio=1.76 (1.3-2.32) at 95% CI]. Th e decrease in systolic blood pressure and diastolic blood pressure after six weeks of treatment of the patients carrying TT genotype were greater than the groups carrying MT and MM genotypes. Th e angiotensinogen (M235T) gene polymorphism is signifi cantly associated with essential hypertension. Patients carrying TT genotype had higher blood pressure lowering response when treated with ACE inhibitor, Enalapril than those carrying MM and MT genotypes suggesting that the T allele may be a possible genetic marker for essential hypertension. Our findings also suggest that C allele of A1166C polymorphism in the angiotensin II type 1 receptor gene is associated with essential
hypertension and its upregulation could play an important role in essential hypertension
Xiping Zhan
Howard University, USA
Title: A reexamination of the role of dopamine receptors in harmaline-induced tremor
Time : 17:10-17:40
Biography:
Xiping Zhan has his expertise in neuropharmacology. His lab uses multiple interdisciplinary approaches to study neural circuits and underlying functional implications. He applies behavior measures to evaluate the neuropharmacology of small molecular drugs on tinnitus, tremor or mood behavior, and uses patch clamping or single unit in vivo recording to address the molecular mechanisms. In addition, he uses human iPS cell derived dopamine neurons to model pharmacology in human. He has been focused on tinnitus and tremor for years in research and teaching
Abstract:
Essential tremor is one of the most common neurological disorders characterized by uncontrollable shaking and tremors throughout the body. Well known to aff ect adults, it can also aff ect children. Harmaline induced tremor is an established animal model for human essential tremor, but its underlying mechanism and eff ects on mood behavior are still elusive. This study aims to use pharmacological and behavioral methods to investigate the pharmacology in harmaline-induced tremor and the auditory startle response. Mice tremors and auditory startle responses were recorded by the Kinder Startle Monitor System. Harmaline (12.5 mg/kg) reliably induced tremor, and that can be attenuated by ethanol (1.5 mg/kg) and sulpiride (20 mg/kg). In addition, it caused the startle response to decrease signifi cantly. Prepulse inhibition and gap responses also decreased upon harmaline injection and increased the following day, but not signifi cantly from the controls. Supplemental administration following recovery can signifi cantly attenuate gap detection without affecting prepulse inhibition. Our data confi rms the frequency of the tremor was from 10 -15 Hz, and the ethanol eff ect, which indicates validity as novel tremor assay. We also found that harmaline attenuates the auditory startle refl ex by causing the reflex and gap detection to be suppressed, but did not aff ect prepulse inhibition signifi cantly. Th ese fi ndings suggest harmaline not only specifi cally modulates sensory-motor integration, but also the timing of gap detection. Our data provides additional information that D2 receptors are involved in harmaline-induced tremor.