Pre Clinical Phase | Safety data generation | Pharmacovigilance Unit 4 Notes and Lecture
Preclinical studies are performed in in vitro, in vivo, ex vivo, and in silico models to obtain basic information about the safety and biological efficacy of a drug candidate before testing it in a final target population, i.e., humans. Preclinical studies or tests are mainly performed in compliance with GLP/GSP guidelines (good laboratory practice and good scientific practices) to ensure reliability and reproducibility of results.
Regulatory aspects of preclinical studies
Preclinical studies mainly follow a combination of:
• GLP (21CFR, 58) and compliance monitoring;
• OECD principles;
• ICH/M3;
• Common Technical Document/CTD module 4 (article 6 of Regulation (EC) No. 726/2004, and with respect to the Annex I to Directive 2001/83/EC);
• Pharmacopoeial or codex requirements;
• 3R principles;
• Local animal ethical committee guidelines and requirements; and
• ISO regulations
Typical tests for chemical entity and pharma drugs.
• Cell viability assays
Thymidine incorporation assays
Cell Titer-Glo cell viability assays
MTS assay
ATPlite cell viability assay
SRB assay
other specific tests
• Cell proliferation assays
MTT cell proliferation assay
cyQuant direct proliferation assay
BrdU cell proliferation ELISA assay
Label-free and real time proliferation assay using IncuCyte
other specific tests
• Cell apoptosis assays
Caspase-Glo 3/7 activation assay
Reaction oxygen species (ROS) assay
Mitochondria membrane potential assay
Apoptosis analysis by flow cytometry
other specific tests
• Cell angiogenesis assay
Matrigel plug assay
Tube formation assay
Platelet activation assay
Co-culture angiogenesis assay
other specific tests
• Related assays
Cell migration assay
Tumor invasion assay
Chemotaxis assay
Cellular phosphorylation assay
Cell cycle assay
Endotoxin assay
Genetic stability testing
Endocrine disruption
Drug-drug interaction
Drug efficacy test
3D angiogenesis assay
3D invasion assay
Inflammation assay
Receptor binding assay
Enzyme inhibition
20 messenger analysis
Plasma stability
Plasma protein binding assay
other specific tests
• In vitro permeability and transporter assays
Caco-2 permeability assay
MDCK permeability assay
Parallel artificial membrane permeability assay (PAMPA)
Transporter assays, e.g., P-glycoprotein (P-gp), BCRP, OCT2
other specific tests
• Type of toxicity assays
2D-based hepatotoxicity assay
3D-based hepatotoxicity assay
Cardiotoxicity
Neurotoxicity
Nephrotoxicity
Genotoxicity
Carcinogenicity
other specific tests
Animal tests
Teratogenicity
Pharmacokinetic
Local tolerance
Pharmacodynamic
Fertility studies
Single dose toxicity in animal
Repeated dose toxicity in animal
other specific tests
Databases
In June 2016, the FDA commissioner proposed building a database that would gather information before clinical trials, meaning from preclinical research (lab animals or cells/cell lines). This proposal of a ClinicalTrials.gov database for preclinical work received mixed reactions from the scientific community. Currently, if a molecule becomes successful, all the drug development data become trade secret; if it does not, the data are stored in archives, and very rarely published despite their high scientific value. The idea of this database supports learning and sharing from unand successful trials.
Even though the idea is interesting, the scientific community is unsure about the future of it, due to several reasons, such as reliability, funding, legal aspects, government and regulatory scrutiny, etc. https://www. preclinicaltrials.eu is trying to promote transparency in preclinical research.
The main objective of this database is to
- increase transparency,
- avoid duplication of animal studies,
- reduce bias reporting,
- increase data sharing on protocols,
- study designs, and outcomes, and foster research collaborations.
The UK’s Catapult database
(https://ct.catapult.org.uk/resources/cell-andgene-therapy-catapult-uk-preclinical-researchdatabase) provides one-stop information for
ongoing preclinical research activity related to cell, gene, and other advanced therapies.
Elsevier’s PharmaPendium database
(https://www.elsevier.com/solutions/pharmapendium clinical-data) collects data on drug safety, the FDA Adverse Event Reporting System (FAERS),and ADME and drug-drug interactions. This database offers risk-benefit analyses and drug candidate assessments.
Researchers can search FDA and EMA regulatory documents, committee meeting minutes, FAERS, data on pharmacokinetics, efficacy, safety, metabolizing enzymes’ and transporters’ potential drug-drug interactions, and guidance on validation of the best animal model. Other databases include http://fprd.ihes.fr (a French preclinical database) and https://dtp.cancer.gov (the NCI’s Development Therapeutics Program, DTP).