GHP April 2017

12 GHP / April 2017 , tested in clinical trials, preclinical studies must unequivocally establish that the Investigational New Drug (IND) is safe to be administered to human subjects in clinical research studies 5 . Specifically, the objective of the preclinical stage of development is to thoroughly profile the toxicity of the IND through experimentation observing good laboratory practices (GLP). Altogen Labs offers in vitro toxicology studies of INDs to characterize the drug candidate’s effects on biological functions and metabolism. Cancer cell lines are used as in vitro models for dose-response studies. Our laboratory houses a large catalog of cancer cell lines to be screened for IC 50 values of candidate drugs. Our research laboratory provides IC 50 studies by screening candidate drugs against our extensive library of cell lines or against client provided custom cell lines. For in vivo research, Altogen Labs offers xenografting – the method of transplanting human tumor tissue into immunocompromised mice to create a more biologically accurate model to investigate an IND’s effect on tumor activity 6 . Altogen Labs offers consultation to our clients on xenograft model selection. Xenograft studies are GLP compliant and animal handling at our facility is IACUC-regulated. Our team of scientists is equipped to perform research studies utilizing more than 80 xenograft models including models for brain carcinoma, pancreatic and breast cancer, epidermoid and nasopharyngeal carcinoma, and xenograft studies for melanoma, colon, breast, lung, and prostate cancers. Additionally, over 20 patient PDX xenograft models are also available. Clients employing xenograft studies are provided with detailed experimental procedures, health reports, and experimental data. These studies can be further expanded to include tissue collection, histology, RNA isolation, and gene expression analysis. Altogen Labs also offers liposome encapsulation services. Liposomes, spherical particles consisting of an aqueous core and at least one lipid bilayer, can be used to encapsulate various types of cargo including DNA, RNA, drugs, and proteins, and serve as efficient delivery vehicles of these agents for in vivo and in vitro studies 7 . Liposome delivery has been established as an effective technique for administration of pharmaceuticals and can be performed topically, orally, or via pulmonary or parenteral routes. Liposomes are specifically advantageous for overcoming solubility issues for difficult-to- formulate agents and are utile for difficult-to-deliver agents like small-interfering RNA molecules. Moreover, liposome encapsulation can augment intracellular transport of drugs and may also facilitate specific delivery of drugs to target cells or tissues. Altogen Labs offers encapsulation of any species of charged molecule (mRNA, siRNA, shRNA, microRNA, plasmid DNA, and protein) into a custom formulation or one of our standard liposome formulations, such as DMPC : DMPG : Cholesterol and PC : DOTAP : PEG : Cholesterol. Microfluidization, extrusion, and sonication methods are available to create a homogenous liposome sample with encapsulated cargo molecules of a specified size (we offer a controlled particle size liposomes of 50 – 400 nm size). Altogen Labs is also experienced in developing and performing in vitro and in situ immuno and cell-based assays. In vitro Enzyme-linked Immunosorbent Assays (ELISAs) are used to characterize antigen or antibody binding activity and are a resourceful tool used in research, diagnostics, and quality control. In situ , cell-based ELISA allows for probing an agent’s effect on cellular signaling pathways and biochemistry. Furthermore, our expertise in cell-based assays grants the ability to expand these studies into cytotoxic and metabolic analysis. Cellular processes that can be elucidated by these means include receptor activation, receptor binding, cell signaling, and ligand internalization. Our lab services include custom ELISA development, optimization, and analysis of customer provided samples via a robust cell-based ELISA, consisting of the compound of interest tested against the activation of 15 Serine/Threonine kinases and 17 Tyrosine kinases. Our team of scientists have years of experience designing unique and novel strategies for immune and cell-based assays. Our analysis techniques and methods include enzyme immunoassay (EIA), Bio-Plex kinase profiling assays, biomarker analysis, receptor binding, cell signaling, cytotoxicity, cell cycle assays, cell viability, and cell proliferation. Furthermore, Altogen Labs can establish a cell-based assay to profile a proprietary compound’s quantitative effects on specific biomarkers, cytokines, or phosphoproteins in cell culture supernatant or whole- cell lysates. In Vivo Toxicology Studies For clients wanting to progress passed in vitro toxicology studies, Altogen Labs also offers in vivo preclinical toxicology studies in rat and mouse models. For preclinical toxicology studies, we offer the options of acute, subchronic and chronic toxicity studies. Acute toxicology studies investigate toxicological effects after a single, large dose of a substance of interest is administered. Longer term subchronic and chronic studies simulate the effects of drugs over a longer course of use and what adverse effects can arise in that timeframe. For subchronic studies, small dosages of the test substance are repeatedly administered over a maximum time period of 90 days. Comparatively, chronic toxicology studies investigate the effects of substances over a longer time course of several months to years. These studies are key in drug development, as they lay the groundwork for further investigation in Phase I clinical trials contingent on the substance yielding favorable results and minimal toxicological, carcinogenic, and mutagenic effects. Additionally, another focus of in vivo toxicology studies is the exploration of how an investigative new drug (IND) is best administered. Delivery of a drug to its target cells in a living system is an intricate process that can be foiled by a myriad of cellular actions. Accordingly, preclinical studies should investigate different drug administration routes to determine how toxicity and efficacy can differ between them. The outcome of these studies should be an optimized procedure for IND administration that is safe, effective, and reduces adverse effects. Complementary to optimizing drug delivery, identification of possible adverse side effects from IND administration is also crucial in attempting to advance the study into clinical trials. Potential side effects can have a deleterious effect on the future health of the subject and/ or its progeny. Potential side effects can be detrimental to the immune system, damaging to the DNA, and harmful to vital organs. Accordingly, additional tests can be performed to study sensitivity, irritation, mutagenic and carcinogenic properties, reproductive toxicity, and immunotoxicity. An INDs application for clinical study can also be augmented by studies of pharmacokinetics/ pharmacodynamics (PK/PD), and absorption, distribution, metabolism, and excretion (ADME). Altogen Labs provides toxicity reports including blood chemistry, markers bioanalysis, pharmacokinetics, pharmacodynamics, and histopathology results. Stable Cell Line Engineering Altogen Labs also offers stable cell line development with a time of completion of 28 days. Access to stable cell