Non-Clinical In-vitro and In-vivo Studies in Drug Development
Abstract
The process of discovering and developing drugs is intricate involving multiple stages of critical importance, from basic research to FDA approval. sequence of steps involved, namely Fundamental Investigation, Preliminary Development, Clinical Testing, and FDA Submission/Authorization. Each stage incorporates major scientific and regulatory assessments, including lead identification, toxicity studies, pharmacological profiling, and clinical testing, spanning Phases I–III. Additionally, Good Laboratory Practice (GLP), Good Manufacturing Practice (GMP), Good Clinical Practice (GCP) at different stages. systematic flow indicates the significance of iterative feedback loops between stages to guarantee safety, efficacy, and regulatory compliance, ultimately leading to the introduction of secure and efficient treatment options to the market.
Keywords:
Lead Optimization, Target Validation, Identification, Mutagenicity, In-vitro Pharmacology, Toxicity, Drug Discovery and Development, Pre-Clinical Development, metabolic stability
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References
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https://www.nature.com/articles/nbt1235
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https://www.researchgate.net/publication/235946588
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doi: 10.4172/1948-5964.1000025
2. Drews J. Drug discovery: a historical perspective. Science 2000;287:1960-1963.
doi: 10.4172/1948-5964.1000025
3. Wang S, Sim TB, Kim YS, Chang YT. Tools for target identication and validation. Curr Opin Chem Biol. 2004;8: 371-777.
doi: 10.4172/1948-5964.1000025
4. Imming P, Sinning C, Meyer A. Drugs, their targets and the nature and number of drug targets. Nature Reviews Drug Discovery. 2006;5:821-834.
doi: 10.22270/ajprd.v7i6.616
5. Croston G. The utility of target-based discovery. Expert Opinion on Drug Discovery. 2017;12(5):427-429.
doi: 10.22270/ajprd.v7i6.616
6. Malviya R, Srivastava M, Bansal V, Sharma PK. Drug design, discovery and development and their safety or efficacy on human body. ResearchGate [Internet]. 2021 [cited 2025 Jun 15]. Available from:
https://www.researchgate.net/publication/356643651....
7. Henning SW, Beste G. Loss-of-function strategies in drug target validation. Current Drug Discovery Technology. 2002;17–21.
Doi: 10.22270/ajprd.v7i6.616
8. John GH, Martyn NB, Bristol-Myers S. High throughput screening for lead discovery. Burgers Medicinal Chemistry and Drug Discovery, 6th edition, Drug Discovery and Drug Development, Wiley Press. 2002;2:37-70.
Doi: 10.22270/ajprd.v7i6.616
9. Patidar AK, Selvam G, Jeyakandan M, Mobiya AK, Bagherwal A, Sanadya G, Mehta R. Lead Discovery and lead optimization: A useful strategy in molecular modification of lead compound in analog design. International journal of drug design and discovery. 2011; 2(2):458-463.
Doi: 10.22270/ajprd.v7i6.616
10. Huber W. A new strategy for improved secondary screening and lead optimization using high-resolution SPR characterization of compound– target interactions. J Mol. Recognition. 2005;18:273–281.
Doi: 10.22270/ajprd.v7i6.616
11. Lofas S. Optimizing the hit-to-lead process using SPR analysis. Assay of Drug Development Technologies. 2004; 2:407-416.
Doi: 10.22270/ajprd.v7i6.616
12. Peixoto MP, Vieira ML, Nunes AS, Araújo SL, Monteiro G. Non-clinical studies in the process of new drug development - Part II: Good laboratory practice, metabolism, pharmacokinetics, safety and dose translation to clinical studies [Internet]. 2016 [cited 2025 Jun 15]. Available from:
https://www.researchgate.net/publication/311567021
13. Friedman LM, Furberg CD, Demets DL. Fundamentals of clinical trials. 4th edition. New York: Springer Science and Business Media LLC; 2010. doi:10.21873/anticanres.13485
14. Van Beijnum JR, Giovannetti E, Poel D, Nowak-Sliwinska P and Griffioen AW. miRNAs: Micro-managers of anticancer combination therapies. Angiogenesis. 2017; 20(2):269-285.
doi: 10.1007/s10456-017-9545-x
15. Phillips RM. Targeting the hypoxic fraction of tumours using hypoxia-activated prodrugs. Cancer Chemother Pharmacol.2016;77(3):441-457.
Doi: 10.1007/s00280-015-2920-7
16. Rhim JS, Tsai WP, Chen ZQ, Chen Z, Van Waes C, Burger AM and Lautenberger JA. A human vascular endothelial cell model to study angiogenesis and tumorigenesis. Carcinogenesis. 1998;19(4):673-681.
17. Filipović L, Aranđelović S, Krivokuća A, Janković R, Dojčinović B and Radulović S. Trans-platinum(II)/(IV) complexes with acetylpyridine ligands as antivascular agents in vitro: Cytotoxic and antiangiogenic potential. Anticancer Agents Med Chem. 2016;16(12):1628-1639.
18. Galloway S.M.. Mut. Res. 1994;312:195-322.
19. Madle S., Korte A. and Ball R. Mut. Res. 1987;182:187-192.
20. Maron D.M. and Ames B.N. Mut. Res.1983;111:173-215.
21. Galloway S.M. Env. Mol. Mutagen. 2000;35:191-201.
22. Sinha S, Roy A, Ghosh M. Unravelling the molecular mechanism of mutagenic factors impacting human health [Internet]. ResearchGate; 2021 [cited 2025 Jun 15]. Available from:
https://www.researchgate.net/publication/354008517
23. Schmid W. Agents Actions.1973;3:77-78.
24. Kirsch-Volders M., Sofuni T., Aardema M. et al. Mut. Res. 2003;540:153-163.
25. Koch MA, Waldmann H. Protein structure similarity clustering and natural product structure as guides to compound library design. In: High-content screening: Science, techniques and applications [Internet]. New York, NY: Springer; 2019 [cited 2025 Jun 15]. p. 137–53. Available from:
https://link.springer.com/protocol/10.1007/978-1-4939-9646-9\_7
26. Taft D, Perry M, La Rocco S, Smith B. Regulatory toxicology and the promise of new approach methodologies (NAMs). Front Toxicol. [Internet]. 2023 [cited 2025 Jun 15];5:1171960. Available from:
https://www.frontiersin.org/articles/10.3389/ftox.2023.1171960/full
27. Guarino RA, editor. New Drug Approval Process: Accelerating Global Registrations. New York: Marcel Dekker, Inc;2004.
Doi: 10.17305/bjbms.2019.4378
28. Darwin C, Wallace J. The Origin of Species. Wordsworth Editions Ltd; 1998
Doi: 10.17305/bjbms.2019.4378
29. Suzanne Clancy. Genetic mutation. Nat Educ. 2008;1:187. Doi: http://dx.doi.org/10.17305/bjbms.2019.4378
30. Oltvai ZN, Barabási AL, Jeong H, Tombor B, Albert R. The large-scale organization of metabolic networks. Nature.2000;407:651-4.
Doi: 10.1038/35036627
31. Jakasa I, Kezic S. Evaluation of in-vivo animal and in-vitro models for prediction of dermal absorption in man. Hum Exp Toxicol. 2008 Apr;27(4):281-8.
doi: 10.1177/0960327107085826.
32. Cressey D. DNA repair sleuths win chemistry Nobel. Nature. 2015 Oct 15;526(7573):307-8.
doi: 10.1038/nature.2015.18515.
33. Lai Y, Chu X, Di L, Gao W, Guo Y, Liu X, Lu C, Mao J, Shen H, Tang H, Xia CQ, Zhang L, Ding X. Recent advances in the translation of drug metabolism and pharmacokinetics science for drug discovery and development. Acta Pharm Sin B. 2022 Jun;12(6):2751-2777. doi: 10.1016/j.apsb.2022.03.009.
34. Wang L, Chiang C, Liang H, Wu H, Feng W, Quinney SK, Li J, Li L. How to Choose In Vitro Systems to Predict In Vivo Drug Clearance: A System Pharmacology Perspective. Biomed Res Int. 2015;2015:857327.
doi: 10.1155/2015/857327.
35. Plant N. Strategies for using in vitro screens in drug metabolism. Drug Discov Today. 2004 Apr 1;9(7):328-36. doi: 10.1016/s1359-6446(03)03019-8.
36. Ma S, Chowdhury SK, Alton KB. Application of mass spectrometry for metabolite identification. Curr Drug Metab. 2006 Jun;7(5):503-23.
doi: 10.2174/138920006777697891.
37. U.S. FDA and the drug development process. Drug Dev Ind Pharm [Internet]. US FDA;2021 [cited 2025 Jun 15]; 47(10):1627–35. Available from:
https://www.tandfonline.com/doi/full/10.1080/03602532.2021.1970178
38. Gómez-Lechón MJ, Donato MT, Castell JV, Jover R. Human hepatocytes in primary culture: the choice to investigate drug metabolism in man. Curr Drug Metab. 2004 Oct;5(5):443-62. doi: 10.2174/1389200043335414..
39. Service RF. A growing role for computational biology in drug development. Nat Biotechnol [Internet]. 2004 [cited 2025 Jun 15];22(10):1235–7. Available from:
https://www.nature.com/articles/nbt1235
40. Singh SS. Preclinical pharmacokinetics: an approach towards safer and efficacious drugs. Curr Drug Metab. 2006;7:165–182.
doi: 10.2174/138920006775541552.
41. Ducharme J, Dudley AJ, Thompson RA. Pharmacokinetic issue in drug discovery. In: Rang HP (Editor), Drug discovery and development. Philadelphia: Churchill Living-stone Elsevier. 2006;141–161.
42. Lee, J. W. et al. Fit-for-Purpose Method Development and Validation for Successful Biomarker Measurement. Pharmaceutical research. 2006;23:312-28.
Doi: 10.1007/s11095-005-9045-3.
43. European Pharmaceutical Review. Drug development and approval processes in Europe [Internet]. ResearchGate [cited 2025 Jun 15]. Available from:
https://www.researchgate.net/publication/235946588
44. Ducharme J, Dudley AJ, Thompson RA. Pharmacokinetic issue in drug discovery. In: Rang HP (Editor), Drug discovery and development. Philadelphia: Churchill Living-stone Elsevier; 2006. P. 141–161.
45. International Conference on Harmonisation (ICH). Q7: Good manufacturing practice guide for active pharmaceutical ingredients; 2000.
46. International Conference on Harmonisation (ICH). M4Q(R1): The common technical document for the registration of pharmaceuticals for human use: quality overall summary of module 2 and module 3; 2002.
47. Types of clinical trials [Internet]. Wikipedia [cited 2025 Jun 16]. Available from:
https://en.m.wikipedia.org/wiki/Clinical\_trial
48. U.S. Food and Drug Administration (FDA). Investigational new drug applications (INDs) – CBER regulated products [Internet]. US FDA; 2025 Mar 27 [cited 2025 Jun 16]. Available from: https://www.fda.gov/vaccines-blood-biologics/development-approval-process-cber/investigational-new-drug-applications-inds-cber-regulated-products
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Mannan A, Rasheed T, Nousheen Z. Non-Clinical In-vitro and In-vivo Studies in Drug Development. Int J Drug Reg Affairs [Internet]. 2025Jun.15 [cited 2026Jan.20];13(2):43-1. Available from: https://www.ijdra.com/index.php/journal/article/view/756
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