GUIDANCE MANUAL FOR ASSURING QUALITY OF TRASTUZUMAB NATIONAL INSTITUTE OF BIOLOGICALS (Ministry of Health & Family Welfare) Government of India A – 32, Sector 62, NOIDA – 201309 Uttar Pradesh – INDIA. E-mail: [email protected] INTRODUCTION Monoclonal Antibodies (mAbs) are immunoglobulin’s (novel protein therapeutics) with a defined specificity derived from a monoclonal cell line. Their biological activities are characterised by a specific binding characteristic to a ligand (commonly known as antigen), and may also be dependent on immune effector function such as antibody-dependent cellular cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC). mAbs may be generated by recombinant DNA (rDNA) technology, hybridomas technology, B lymphocyte immortalisation or other technologies (e.g. phage display technology, genetically engineered animals). As a result of naturally-occurring molecular heterogeneity, imperfect cellular processing, chemical and enzymatic changes during manufacturing and additional changes upon storage, antibody drugs display a wide variety of minor chemical changes, collectively termed microheterogeneity. Common examples include glycan structural differences, deamidation, oxidation and glycation. Control of microheterogeneity within predefined analytical specifications has been used in quality control laboratories to guarantee consistent product quality during cGMP manufacturing. The recent Quality by Design (QbD) initiative for therapeutic biotechnology products, a joint pilot program between the regulators, academia and the biotechnology industry, is providing new guidance and expectations on QbD approaches in manufacturing for therapeutic proteins. As the name indicates, QbD encourages developers to build quality into the drug from the start. This approach requires significant knowledge of the drug’s mechanism of action and how the drug’s attributes affect quality. Physical or chemical changes known to affect the safety or efficacy of the drug are considered Critical Quality Attributes (CQAs). Manufacturing is then designed to control the desired levels of CQAs within defined limits, providing a consistent product quality. With this new QbD paradigm, the process is defined by the target ranges for the CQAs which, in turn, provide assurance of consistent product quality. Of the various classes of antibodies, or immunoglobulin’s, IgG1 is the most common immunoglobulin used for pharmaceutical and biomedical purposes, however, recent developments have shown that other immunoglobulin types (e.g., IgG2, IgG4) and mAb-related products (e.g., Fc-fusion proteins, Fabs, etc.) are also being used for therapeutic purposes. The technological advancement in the evolution of therapeutic antibodies is shown at Figure – 1. Figure - 1: Various types of mAbs evolved with technological advancement. The innovator product of trastuzumab is a humanized IgGl kappa monoclonal antibody that selectively binds with high affinity to the extracellular domain of the human epidermal growth factor receptor 2 protein, HER2 (Figure – 2). Trastuzumab is produced by recombinant DNA technology in a mammalian cell (Chinese Hamster Ovary) culture. It was the first targeted cancer drug, aimed at patients whose tumors have specific genetic characteristics. In this case, the appropriate patients have tumors that over-express the HER2 gene. Antigen binding sites: derived from mouse Figure - 2Human IgG1 Antibody Trastuzumab is mainly use for breast cancer treatment. Breast cancer is the most common cancer among women worldwide. Each year, about 1.4 million new cases of breast cancer are diagnosed worldwide, and over 450,000 women will die of the disease annually.In India Breast cancer is the second most frequently diagnosed cancer (22%) and is the second leading cause of death after cervix cancer (26%) among women. In October 20, 2010, the U. S. Food and Drug Administration granted approval for trastuzumab (Herceptin®, Genentech, Inc.), in combination with cisplatin and a fluoropyrimidine (capecitabine or 5-fluorouracil), for the treatment of patients with HER2 overexpressing metastatic gastric or gastroesophageal (GE) junction adenocarcinoma, who have not received prior treatment for metastatic disease. In HER2-positive breast cancer, increased quantities of the human epidermal growth factor receptor 2 (HER2) are present on the surface of the tumor cells. This is known as "HER2 positivity" and affects approximately 15% to 20% of women with breast cancer. HER2-positive cancer is a particularly aggressive form of breast cancer. 2. SCOPE The guidance manual is aimed at emphasizing on the quality control testing parameters particularly for trastuzumab intended for therapeutic use. Parallel to the aforesaid, there is a need felt to meet the requirements of good manufacturing practices, product-lot-testing to ensure product reproducibility, production process verification and quality assurance. 3. GLOBAL AND INDIAN MARKET DYNAMICS OF MAb PARTICULARLY TRASTUZUMAB The recombinant humanized anti-Her2 antibody trastuzumab (Herceptin) from Genentech/Roche is among the world's commercially most successful antibodies (2011 sales of over US$ 5.5 bn). Herceptin is approved and marketed for Her2 positive breast and gastric cancer. The commercial attractiveness of trastuzumab has stimulated many companies to create and develop next generation anti-Her2 monoclonal antibodies (mabs) with improved properties compared with the humanized antibody trastuzumab. Accordingly, Genentech and Roche are still expanding the indication label of trastuzumab and have developed 2nd and 3rd generation Her2 antibodies. These Biosuperior antibodies include antibody-drug conjugates, bispecific antibodies and engineered Her2 antibodies. The forthcoming patent expiry for Herceptin has boosted interest in and activities for developing biosimilar copies with the most advanced projects already in phase III. At present, there are at least 7 Herceptin biosimilar antibodies in clinical development and further 15 in preclinical R&D. As the patents for biologics is expired or move closer to their expiration dates and the challenging economic situation across the globe forces a decrease in healthcare expenditures, biosimilars emerge as the cost-effective therapeutic alternative across the globe. Market for biosimilar mAbs is on the constant rise. The average patent expiry of $39.6 bn per annum between 2010 to 2015 as compared to just $14.2 bn in last decade and 16.5 bn per annum between 2016-2020 (Table -1) Table -1 Being an affordable yet equally-effective alternative for biologics, they can provide savings that range from 25% to 85% of the original versions. Moreover, the patent expiry of approximately 48 biologics (worth US $73 billion) in the next 10 years will also fuel the demand for biosimilars. Analysts estimate that biosimilars could quickly become a $10 billion category (Table – 2 & 3). Despite the resistance from the innovator companies, and the regulatory hurdles involved in their production, biosimilars have immense market potential for both developed, as well as growing economies. Table – 2: FDA approved therapeutic mAbs available in the world market. Name: Antibody *OKT3: Muronomab-CD3 Target: Antibody Type CD3: Murine, IgG2a Indication Company ReoPro: Abciximab PIIb/IIIa: Chimeric, IgG1, Fab Rituxan: Rituximab CD20: Chimeric, IgG1 *Zenapax: Daclizumab CD25: Humanized, IgG1 Autoimmune Roche Simulect: Basiliximab CD25: Chimeric, IgG1 Autoimmune Novartis Synagis: Palivizumab RSV: Humanized, IgG1 Infections MedImmune Remicade: Infliximab TNFa: Chimeric, IgG1 Autoimmune Johnson & Johnson Herceptin: Trastuzumab HER2: Humanized, IgG1 Cancer Genentech/ Roche *Mylotarg: Gemtuzumab ozogamicin CD33: Humanized, IgG4, immunotoxin Cancer Wyeth/ Pfizer Campath: Alemtuzumab CD52: Humanized, IgG1 Cancer Genzyme Zevalin: Ibritumomab tiuxetan CD20: Murine, IgG1, radiolabeled (Yttrium 90) Cancer Humira: Adalimumab TNFa: Human, IgG1 Autoimmune Abbott Xolair: Omalizumab IgE: Humanized, IgG1 Autoimmune Genentech/ Roche Approval Date Autoimmune Johnson & Johnson 1986 (US) Homeostasis Johnson & Johnson 1984 (US) Cancer Genentech Patent Expiry Data Not available 1997 (US) Data Not available 26.11.2017 1998 (EU) 02.06.2018 1997 (US) 1998 (US) Data Not available Data Not available Data Not available Data Not available Data Not available Data Not available 24.08.2018 1999 (EU) 13.08.2019 1999 (EU) 1998 (US) 1998 (EU) 1998 (US) 1999 (EU) 1998 (US) 2000 (EU) 25.09.2018 28.08.2018 2000 (US) Data Not available 2001 (US) 07.05.2021 06.07.2021 2001 (EU) 2002 (US) Biogen Idec 2004 (EU) 2002 (US) 2003 (EU) Data Not available Data Not available 31.12.2022 08.09.2023 20.06.2023 2003 (US) Bexxar: Tositumomab-I-131 *Raptiva: Efalizumab CD20: Murine, IgG2a, radiolabeled (Iodine 131) CD11a: Humanized, IgG1 Cancer Autoimmune Corixa/GSK 2003 (US) 2003 (US) Data Not available 2004 (EU) Data Not available Genentech/ Roche 2004 (US) Erbitux: Cetuximab EGFR: Chimeric, IgG1 Cancer Imclone/ Lilly Avastin: Bevacizumab VEGF: Humanized, IgG1 Cancer Genentech/ Roche Tysabri: Natalizumab a4-Intergrin: Humanized, IgG4 Autoimmune Biogen Idec 2004 (US) Actemra: Tocilizumab Anti-IL-6R: Humanized, IgG1 Autoimmune Chugai/ Roche 2005(JP) 2010 (US) Vectibix: Panitumumab EGFR: Human, IgG2 Cancer Amgen 2006 (US) Lucentis:Ranibizumab VEGF: Humanized IgG1 Fab Macular degeneration Genentech/ Roche Soliris: Eculizumab C5: Humanized IgG2/4 Cimzia: Certolizumab pegol TNFa: Humanized, pegylated Fab Autoimmune UCB 2008 (US) Simponi: Golimumab TNFa: Human IgG1 Autoimmune Johnson & Johnson 2009 (US, EU, CAN) Ilaris: Canakinumab IL1b: Human IgG1 Infalmmatory Novartis 2009 (US,EU) Stelara: Ustekinumab IL-12/23: Human IgG1 Autoimmune Johnson & Johnson Arzerra: Ofatumumab Prolia: Denosumab Numax: Motavizumab ABThrax: Raxibacumab Benlysta: Belimumab Yervoy: Ipilimumab Adcetris: Brentuximab Vedotin Perjeta: Pertuzumab Blood disorders Alexion 2004 (EU) 2004 (US) 2005 (EU) 2006 (US) 2007 (US) 2009 (US) 2008 (EU) CD20: Human IgG1 Cancer Genmab 2009 (EU) RANK ligand: Human IgG2 Bone Loss Amgen 2010 (US) RSV: Humanized IgG1 Anti-infective Meddimmune Pending B. anthrasisPA: Human IgG1 Anti-infection GSK 2012(US) BLyS: Human IgG1 Autoimmune Human Genome Sciences 2011 (US) CTLA-4: Human IgG1 Cancer BMS 2011 (US) CD30: Chimeric, IgG1, Drug-conjugate Her2: Humanized, IgG1 Cancer Cancer Seattle Genetics Genentech/ Roche Data Not available 2011 (US) 2012 (US) 12.02.2024 Data Not available 26.02.2024 Data Not available Data Not available Data Not available Data Not available Data Not available Data Not available Data Not available Data Not available Data Not available Data Not available Data Not available Data Not available Data Not available Data Not available Data Not available Data Not available Data Not available Data Not available Data Not available Kadcyla: Ado-trastuzumab emtansine Her2: Humanized, IgG1, Drug-conjugate Cancer Genentech/ Roche Data Not available 2013 (US) *Withdrawn by the sponsor Table 3: List of biologics coming off patent from 2012 to 2015. Product Company Therapeutic Category Patent Expiry Enbrel Amgen, Pfizer Other anti-rheumatics 23-10-2012 Neupogen Amgen Immunostimulants 12-03-2013 Humalog Eli Lilly Anti-diabetics 07-05-2013 Avonex Biogen Idec MS Therapies 30-05-2013 Epogen Amgen Anti-anaemics 20-08-2013 Procrit/Eprex Johnson & Johnson Anti-anaemics 20-08-2013 Cerezyme Genzyme Other therapeutic products 27-08-2013 Rebif Merk KGaA MS Therapies 21-12-2013 Novomix Novo Nordisk Anti-diabetics 06-06-2014 NovoRapid/NovoLog Novo Nordisk Anti-diabetics 07-12-2014 Rituxan Roche Antineoplastic Mabs 31-12-2014 Kogenate Bayer Anti-fibrinolytics 31-12-2014 Prevnar Pfizer Vaccines 01-01-2015 Lantus Sanofi-Aventis Anti-diabetics 12-02-2012 Actemra Roche Other Anti-rheumatics 07-06-2012 Gonal-F/Gonalef Merk KGaA Fertility agents 16-06-2015 Neulasta Amgen Immunostimulants 20-10-2015 Nimotuzumab YM BioSciences Anti-neoplastic Mabs 17-11-2015 Norditropin SimpleXx Novo Nordisk Growth Hormones 15-12-2015 Helixate CSL Anti-fibrinolytics 31-12-2015 3.1 Indian Market: India, with a share of over 2 per cent in the global biotechnology industry, is set to become one of the world's largest biotechnology hubs. Nearly 64 per cent of the biotech companies operate in the bio-pharma sector in India. The biotechnology sector in India is expected to achieve revenue of US$ 11.6 billion by 2017, growing at a compound annual growth rate (CAGR) of 22 per cent. Revenue from biotech exports reached US$ 2.2 billion in FY13, accounting for more than half (51 per cent) of total industry revenues. Biotech industry is expected to grow to around Rs 440,000 crores (US$ 70.84 billion) by the year 2020.The vaccines and recombinant therapeutics are the leading sectors driving the biotechnology industry's growth in India. M/s Biocon, other Indian pharma companies like M/s Intas, M/s Mabpharma and M/s Dr. Raddys Lab are also set to tap the Rs.130-crore market for breast cancer drug Trastuzumab. Just five top Indian companies with manufacturing capabilities in recombinant therapeutic products in the year 2005 are shown in Table – 4. Table 4: 3.2 Biosimilar Therapeutics - Indian Scenario: The drug classes for biosimilars in India comprise of Human Insulin, Human Growth Hormone, Granulocyte Colony Stimulating Factor (G-CSF), Erythropoietin, and Streptokinase. Recent estimates indicate that 20 companies in India are already developing biosimilars. There are around 25 recombinant therapeutic available in India and 15 of them are manufactured within the country. About 50 brands are commercialized in the country by leading companies such as Biocon, Shantha Biotechnics, Reliance Life Sciences, Wockhardt and Intas Biopharmaceuticals among others. About 72 recombinant drugs are currently undergoing different stages of clinical trials and their launch may cause an increase of 25% in the Indian Biopharma sector. In the biosimilar market India was holding 26th position in 2005 and will be expected to hold 6th position by 2015 in the global biosimilar market (Figure - 3) Figure – 3 4. ICH DOCUMENTS FOR BIOLOGICS: 5. MANUFACTURING PROCESS (UPSTREAM & DOWNSTREAM) 6. STAGES OF MASTER CELL BANK (MCB) AND WORKING CELL BANK (WCB). 7. QUALITY CONTROL: 7.1 Reference Material There is a need to have reference standard of Trastuzumab. In the meantime innovator drug is regularly used as a comparator as and when required for quality control testing. 7.2 Lot release testing of Trastuzumab Once the trastuzumab is purified and formulated, the resulting drug substance must be tested prior to lot release (Table – 5). The product specific specifications are not available in Pharmacopoeia. Therefore lot release of drug is done based on the limits approved for the drug during the marketing authorization by National Regulatory Authority (NRA). A set of tests and acceptance criteria are established based on trastuzumab characterization and prevalent regulatory requirements in order to ensure drug quality. These tests typically include appearance, identity, purity, protein concentration, potency of the drug, microbial limits or bio-burden, bacterial Endotoxins, Host cell protein and residual DNA etc. Ion exchange chromatography, SDS under reducing and non-reducing condition by Capillary Electrophoresis and Size Exclusion Chromatography are the most frequently used lot release methods for purity for Trastuzumab. Test for immunogenicity is performed by the manufacturer. EMA has prepared a concept paper to revise the guidelines on immunogenicity assessment on 20th February 2014. The aim of reform is not to increase the number of studies on immunogenicity but to increase the quality of studies and their clarity to the assessors. Stability studies may be performed on the Trastuzumab drug substance (e.g. frozen bulk for storage) and drug product, if required (e.g. final vial) according to prevalent regulatory guidelines. EMEA 2006 & 2007 guidelines are available for characterisation of similar biological medicinal products containing biotechnology derived proteins as active substances. WHO (2013) guidelines have been published on evaluation of similar biotherapeutic products. Similarly, guidelines on similar biologics; regulatory requirements for marketing authorization in India have been published in the year 2012 by the DBT & CDSCO, Govt. of India. Indian pharmacopoeia commission is in the process of finalisaiton of monograph on Rituximab and other recombinant drugs. Table – 5. Commonly used tests in a Certificate of Analysis for lot release of mAbs including trastuzumab. QC Parameter Test Name Appearance Color, Opalescence and Clarity Identity Peptide mapping by RP-HPLC (Reverse Phase HPLC), or MALDI (Matrix – assisted deionisation) Mass spectroscopy, or UV spectroscopy (2nd derivative) Purity Limulus Amebocyte Lysate (Endotoxin) Size exclusion chromatography (SEC) CE-SDS (Capillary electrophoresis-Sodium dodecyl sulphate) IEC (Ion exchange chromatography) or icIEF (Imaged capillary isoelectric focusing) Glycosylation profile Peptide mapping by RP-HPLC Potency Potency (ELISA/Cell-Based Assay) Strength UV Spectroscopy General Tests Osmolality pH Protein concentration Surfactant Concentration (e.g. Polysorbate 20) Safety Sterility testing Bacterial Endotoxin Test Critical Quality tests such as Identity, Purity, Potency and Safety are performed head to head with the “Reference” by using test methods suitably approved by a National Control Authority. 7.3 General Test: Appearance, Solubility, pH, Osmolality, Water content, Protein content, Subvisible particulate matter and Extractable volume should be assessed wherever appropriate as per the prevalent regulatory guidelines / pharmacopoeia. The results complies with the limits approved for the particular product during marketing authorization. 7.4 Identification: A. By Peptide Mapping: Positive identity is confirmed if the elution profiles of the test solution and reference solution are similar by visual observation and there are no new significant peaks. It complies with the limits approved for the particular product during marketing authorization. B. By Iso Electric Focussing (IEF) using capillary electrophoresis (CE): The identity is established comparing the product with reference preparation. It complies with the limits approved for the particular product during marketing authorization. C. CZE by capillary electrophoresis: The difference in the migration time (MT) for the main peak of the first and second reference solution injections bracketing each test solution must be < 0.1 minute. It complies with the limits approved for the particular product during marketing authorization. D. Bio-assay: Same as potency assay 7.5 Impurities: A. Size Exclusion Chromatography by HPLC (SEC-HPLC): Impurities are determine by Size Exclusion Chromatography and results complies with the limits approved for the particular product during marketing authorization B. Ion Exchange Chromatography by HPLC (IEC-HPLC): Impurities are determined by Ion Exchange Chromatography. Run the sample with appropriate gradient program to optimize the separation. It complies with the limits approved for the particular product during marketing authorization. C. SDS by Capillary Electrophoresis (CE-SDS): Impurities are determined by Capillary Electrophoresis. This test performed in under reducing as well as nonreducing condition. Analyze the sample and reference against the molecular weight marker using an capillary electrophresis method capable of giving separation in the range 10- 250 kDa followed by UV detection. Results complies with the limits approved for the particular product during marketing authorization. 7.6. Safety Test: A. Sterility: It complies with the test for sterility B. Bacterial Endotoxin Test (BET): Not more than 1.0 EU per mg. 7.7. Assay: 1. Potency (Anti-proliferation): Determine the potency of Trastuzumab by using BT-474 or similar cell line in an antiproliferation assay. It complies with the limits approved for the particular product during marketing authorization. 2. Glycan Profilling by Capillary Electrophoresis or by HPLC: By any one of validated methods. It complies with the limits approved for the particular product during marketing authorization. 8. REFERENCES: I. A Report by Association of Biotechnology Led Enterprises (ABLE) for the Department of Biotechnology, Government of India – May 2012. II. Biospectrum – Volume 9, Issue 6, June 2011. III. EMEA guidelines on similar biologicals medicinal products containing biotechnology derived proteins as active substance: non-clinical and clinical issues, London – 2006 (CHMP/BMWP/42832). IV. EMEA guidelines on immunogenicity assessment of biotechnology derive therapeutics proteins, London – 2007 (CHMP/BMWP/14327). V. Guideline on similar biologics – Regulatory requirements for marketing authorisation in India (2012), Department of Biotechnology, Ministry of Science & Technology, Government of India, CDSCO – Ministry of Health & Family Welfare, Government of India (Page No’s: 1 to 42) VI. VII. ICH document – Manufacturing process of Biologics - K. Ho Afssaps, France (2011). Monoclonal Antibody Development and Physicochemical Characterization Jennifer C. Rea, Yajun Jennifer Wang, Tony G. Moreno, Rahul Parikh, Yun Lou and Dell Farnan, Genentech, Inc., Protein Analytical Chemistry, South San Francisco, CA-USA – 2012. VIII. WHO-TRS 814 (1991), Annex 3 – Guideline for assessing the quality of pharmaceutical & biotech products prepared by rDNA technology. IX. WHO guideline – “Guidelines for national authorities on quality assurance for biological products” – WHO TRS 822 (Annex 2) – 1992. X. WHO Guidelines on Biosimilars evaluation of Similar Biotherapeutic products (SBPs) – 2009. XI. WHO informal consultations on the revision of guidelines on the Quality, Safety, Efficacy of Biological & Medicinal products prepared by rDNA technology- June 2012. XII. Anti-malignant Tumor Agent with Anti-HER2 Humanized Monoclonal Antibody: Presentation of Takahiro Yamaguchi, Herceptin Product Manager, Chugai Pharmaceutical Co., Ltd. Oncology Unit, Breast Cancer Group. XIII. From pipeline to market R&D Directions 2009, 15(6)4-89. XIV. http://photos.prnewswire.com/prnh/20100302/LA63139LOGO). XV. http://www.researchandmarkets.com/research/jdh26g/competitor. XVI. http://www.businesswire.com/news/home/20130402005802/en/ResearchMarkets-Competitor-Analysis-Her2-Antibodies. Disclaimer: This guidance represents the current views of National Institute of Biologicals (NIB's) on assuring Quality of Therapeutic (MAbs). It does not create or confer any rights for or on any person and does not operate to bind NIB or the public. An alternative approach may be used if such approach satisfies the requirements of the applicable statutes and regulations.
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