1. CHALLENGES FACED IN THE DEVELOPMENT OF BIOSIMILARS Dr.G.Hima Bindu MD; PG dip. diabetology Asst.Professor Dept. of Pharmacology Rajiv Gandhi Institute of Medical Sciences Ongole drgujjarla@gmail.com

2.  Biologics  Differences  Manufacturing process  Challenges in the development 28.10.20142

3. INTRODUCTION Biologics derived from living cells Currently used in  Blood conditions : leuco/neutro/pancytopenia  Cancer: colon, breast, non Hodgkins lymphoma  Immune system disorders: RA, psoriasis  Neurological: multiple sclerosis World wide, nearly 200 biologics saved over 800 million patients 28.10.2014 3

4. Need for biosimilars Increasing demand for biologics  Patent expiry  Takes more time and cost to develop Alternate version of innovator biopharmceuticals – Similar biotherapeutic products : WHO – Biosimilars : Europe – Follow – on – biologics : US 28.10.20144

5. 5 Biosimilars are similar……. …. Not Identical WHO: “A biotherapeutic product similar in terms of quality, safety and efficacy to an already licensed reference biotherapeutic product.” Similarity must be shown on the basis of the analytical, non clinical and clinical data

6. DIFFERENCES 28.10.20146

7. 7 Chemical drugsBiologics Small, simple molecule (mol. Wt 100 -1000Da) with defined structure Large complex molecules (mol.wt 15000 -150000Da), Heterogenous structure Made by chemical synthesis Made by living cells Easy to characterizeDifficult to characterize Easy to purifyLengthy and complex purification process

8. 28.10.20148 Contamination avoided, easily detectable & removable High possibility of contamination. Detection is harder Relatively stable Variable, sensitive to environmental conditions Administered by various routes Parenteral Non immunogenicImmunogenic Chemical drugsBiologics

9. MANUFACTURING PROCESS 28.10.20149

10. Manufacturing Process Complex process Developed through sequential process Systematically engineered to match the reference product 28.10.201410

11. Major steps involved in the manufacture 28.10.201411 Gene selection Insertion into vector Cell culture Protein productionPurification Formulation

12. CHALLENGES 28.10.201412

13. Cell line generation Protein production Protein purification 28.10.201413

14. Microbial cells, cell lines of human or animal origin or tissues derived from animals or plants are used. Bacteria & yeast cell lines require minimal growth media conditions and are fast growing.  Traditional cell lines are prone to host cell protein contamination. 28.10.201414 Cell Line Generation

15. Cont…. 28.10.201415 Chinese Hamster ovary (CHO) cells are preferred mammalian cell line If the host cell line used for production of reference product is known same cell line is used.  The expression system can have a significant effect on the types and extent of translational and post translational modifications.

16. 28.10.201416 Every manufacturer uses unique cell line and proprietary process. So it is difficult to produce biosimilars that are identical to the original drug. Cont….

17. Protein Production 28.10.201417 Structure of protein is heterogeneous - exhibit complex three dimensional conformations.  The structure – function relationships are very sensitive as modifications of may affect safety, purity and/ or potency.  ELISA and size exclusion chromatography are used

18. 28.10.201418  The function of a protein can be altered by Replication errors in the DNA encoding protein sequence Amino acid misincorporation during translation Post translational modifications Cont….

19. 28.10.201419 Protein PhosphorylationGlycosylation Lipid attachmentProteolytic cleavage Increase half life Membrane stability Activation or inhibition Activation Post translational modifications

20. 28.10.201420  Changes in protein modification may reduce biological activity  These post translational modifications are affected by the cell line.  Glycosylation is sensitive to cell growth conditions  changes in culture pH  the availability of precursors and nutrients  presence or absence of various cytokines and hormones. Cont….

21. 28.10.201421  It is difficult to characterize glycosylation pattern or the higher order/ 3D structure.. Tools to detect:  SDS-PAGE  RP-HPLC  Size exclusion chromatography  Mass spectroscopy Cont….

22. Protein Purification 28.10.201422  Oxidation/deamidation or even protein aggregation can occur during the process.  Protein molecules can also be degraded and impurities created Additives used to stabilize proteins: – human serum albumin (HSA), – polysorbates, – calcium chloride

23. Immunogenicity 28.10.201423 All therapeutic proteins induce some antibody response. Various factors influence immunogenicity patient related characteristics  Product related factors

24. 28.10.201424 Molecule design Cell line Deglycosylation Exposure of antigenic sites Misfolding/ aggregation Impurities Product related factors

25. 28.10.201425 The increasing demand for more cost effective treatment and patent expiry of biologics created growing market for biosimilars. It is difficult to develop biosimilars identical to original drug. Use of different vector Diff. Cell expression system Diff. in growth media, nutrients Diff. operating methods, reagents, reference products SUMMARY

26. 28.10.201426