Bio-similar & biobetter mAbs: Characterisation, comparability & adverse events

Roy Jefferis School of Immunity & Infection, University of Birmingham UK

GBX Summit, London, February 10-11th 2015

The first area where further innovation should be expected is product quality and productivity for upstream processes. Given the inherent complexity of biological systems any enhancement in productivity needs to be tempered by a rigorous evaluation of product attributes and quality. Although upstream production uses mature technology, intracellular processes involved in product translation, folding, post-translational modifications and secretion are still only partly understood, which would suggest there is room for further improvement.

The need for innovation in biomanufacturing.

Gottschalk U. et al. Nature Biotechnology, 30:489–492 (2012)

The “biggest change” to a therapeutic protein would be the independent development of a manufacturing process by a new manufacturer e.g. a biosimilars (i.e., a “copy” version of an already licensed originator product). EU requirements: A biosimilar mAb should be similar to the reference mAb in physicochemical and biological terms. Any observed relevant difference would have to be duly justified and could contradict the biosimilar principle.

Comparability exercise and biosimilars

http://www.ema.europa.eu/docs/en_GB/document_library/Scientific_guideline/2010/11/WC500099361.pdf

http://www.ema.europa.eu/docs/en_GB/document_library/Scientific_guideline/2009/09/WC500003074.pdf;

245 companies developing or already selling biosimilars 700 biosimilars approved or moving through pipeline Biosimilars sell at 20 – 30 % discount In total biosimilars are expected to account for ~ 25 % of the $100 billion worth of sales stemming from off-patent biological drugs by the end of the decade

Commercial biosimilar activity 2014:

http://www.reuters.com/article/2014/09/29/health-biotech-biosimilars-idUSL6N0RG2MA20140929

Product Biosimilars Biobetters

Humira (adalimunmab) 20 7

Remicade (infliximab) 13 8

Epogen (epoetin alfa) 82 25

Neupogen (filgrastim) 56 15

Enbrel (etanercept) 26 11

Rituxan (rituximab) 43 19

Herceptin (transuzumab) 37 14

Biosimilars and biobetters in development

Citation: Clinical Pharmacist, 6: November 2014; online | URI: 20067091

In 2009 14 biosimilar EPOs were licensed in Thailand; manufactured in Argentina, China, South Korea, and India. Concomitant with the increased penetration of biosimilars the authors noted an alarming increase in the prevalence of pure red-cell aplasia (PRCA). They were able to confirm that 23 of 30 patients referred because of loss of ESA efficacy had antibodies to erythropoietin, consistent with PRCA.

Wish JB. Kidney International 80, 11–13 (2011)

Erythropoiesis stimulating agents (ESA)

Product % Ab incidence

Humira 5 - 89

Remicade (CD) 61

Remicade (RA) 21

Campath-1H 63

GM-CSF (1) 74

GM-CSF (2) 95

IL-2 53

Immunogenicity of biopharmaceuticals:

Guidance for Industry: Immunogenicity Assessment for Therapeutic Protein Products

…. it should be noted that chimeric, humanized and human monoclonal antibodies can also elicit a high rate of immunogenicity depending on the dosing regimen and patient population. … novel structural formats, including fusion proteins, ADC, bispecific or multispecific, specifically engineered antibodies etc. may elicit immune responses, as such novel structures may create neoantigens or expose cryptic epitopes.

http://www.fda.gov/downloads/drugs/guidancecomplianceregulatoryinformation/guidances/ucm338856.pdf

Sales 2012 - blln Herceptin (trastuzumab) – EU 2014, US 2019. $6.8 Humira (adalimumab) – EU 2018, US 2016. $9.26 Remicade (infliximab) – EU 2014, US 2018. $8.2 Rituxan (rituximab) – EU 2013,US 2018. $6.9

http://www.totalbiopharma.com/2013/05/07/biosimilar-developers-due-cash-5-therapeutic-mabs/

Patent expiry dates

The Patient Protection and Affordable Care Act

Amends the Public Health Service Act (PHS Act) to create an abbreviated licensure pathway for biological products that are demonstrated to be “biosimilar” to or “interchangeable” with an FDA-licensed biological product. This pathway is provided in the part of the law known as the Biologics Price Competition and Innovation Act (BPCI Act).

Signed into law by President Obama on March 23, 2010

Under the BPCI Act, a biological product may be demonstrated to be “biosimilar” if data show that, among other things, the product is “highly similar” to an already-approved biological product.

The Patient Protection and Affordable Care Act

http://www.fda.gov/drugs/developmentapprovalprocess/howdrugsaredevelopedandapproved/approvalapplications/therapeuticbiologicapplications/biosimilars/default.htm

CH3

VH

CL

VL

CH2

CH1

--S-S-- ----S-S----- ----S-S-----

Heavy chain: ~ 440 amino acids

Light chain: ~ 210 amino acids

Basic structure of an IgG antibody molecule

Antigen

binding

Fc Effector functions

Ludger

Domain structure of IgG

Fab Fab

Antigen

binding

oligosaccharide

Jefferis R. Arch. Biochem. Biophys. 526:159-662 (2012)

Ligands mediating IgG-Fc effector functions

FcγRI (IgG1,3,4) FcγRIIa* (IgG1,2,3) FcγRIIIa* (IgG1,3,4*)

phagocytosis, antibody dependent cellular cytotoxicity (ADCC), generation of superoxide, release of enzymes, immunoregulation

* dependent on FcγR polymorphisms; * dependent on glycoform

Complement: complement dependent cytotoxicity (CDC)

C1q (IgG1,3) Classical pathway

MBL (IgG1,2,3,4) Lectin pathway

FcRn (1,2,3,4) Catabolism & placental transport

Jefferis R. Arch Biochem Biophys 526:159-166 (2012) Lejeune J., Watier H et al., Bull Cancer 97:511-522 (2010)

CH3

VH

CL

VL

CH2

CH1

--S-S-- ----S-S----- ----S-S-----

- glycosylation

- deamidation

- deimination

- Met oxidation in the Fc

- C-terminal Lys processing

- hinge-region fragmentation

- glycation of Lys residues

- Pyro-Glu

Post translational and chemical modifications

Kozlowski, S. et al. Adv. Drug Delivery Reviews 58:707–72 (2006) Harris RJ. Dev Biol (Basel) 122:117-27 (2005)

Heterogeneity: 108 molecular forms possible

Transgenics: goat; sheep; cows; rabbits; pigs etc

Aves: chickens (eggs)

Yeasts: Pichia pastoris; Saccharomyces cerevisiae

Insect cells: Sf9 (baculovirus infected)

Plants: tobacco; corn; tomato; potato; moss

Bacteria: Escherichia coli; Bacillus subtilis

Glycoprotein production vehicles:

Mammalian: CHO, Sp2/0; NSO; Per.C6; HEK 293 etc

Liberia to receive Zmapp drug to treat Ebola virus

Ebola Drugs (ZMapp) produced in tobacco leaves

A Spanish priest infected with Ebola flown home for treatment

Murin CD. et al. Proc Natl Acad Sci U S A. 111:17182-7 (2014).

http://time.com/3457472/see-how-ebola-drugs-grow-in-tobacco-leaves/#3457472/see-how-ebola-drugs-grow-in-tobacco-leaves/

All product applications should contain a complete and thorough chemistry, manufacturing and controls (CMC) section that provides the necessary and appropriate information Manufacturers should provide to FDA extensive chemical, physical and bioactivity comparisons with side-by-side analyses of the "old" product and qualification lots of the "new" product. When available, fully characterized reference standards for drug substance and final container material should also be used.

http://www.fda.gov/Drugs/GuidanceComplianceRegulatoryInformation/Guidances/ucm122879.htm

Demonstration of Comparability of Human Biological Products, Including Therapeutic Biotechnology-derived Products

Physicochemical assessment of the proposed biosimilar product and the reference product should consider all relevant characteristics of the protein product (e.g., the primary, secondary, tertiary, and quaternary structure, post-translational modifications, and functional activity(ies)). The objective of this assessment is to maximize the potential for detecting differences in quality attributes between the proposed biosimilar product and the reference product.

http://www.fda.gov/downloads/drugs/guidancecomplianceregulatoryinformation/guidances/ucm291134.pdf

Demonstration of Comparability of Human Biological Products, Including Therapeutic Biotechnology-derived Products

Korean Ministry of Food & Drug Safety

Remsima (remicade)

The first biosimilar antibody therapeutic is approved by the EMA

“Remsima is a biological medicinal product similar to the reference product Remicade (infliximab) authorised in the European Union since August 1999. Studies have shown Remsima to have a comparable quality, safety and efficacy profile to Remicade (infliximab). Remsima is approved for all indications for which the innovator product Remicade (Infliximab) is approved.

http://www.ema.europa.eu/docs/en_GB/document_library/Scientific_guideline/2009/09/WC500003953.pdf

http://www.ema.europa.eu/docs/en_GB/document_library/Scientific_guideline/2009/09/WC500003074.pdf;

Approval for Remsima demonstrates that the EMA has confidence in its ability to evaluate the comparability of biosimilar antibody products. The successful development and approval of Remsima, following EMA guidelines, may act as an incentive for the biosimilar “industry” and establishment of definitive FDA guidelines.

Blogg

Approval by the EMA as a landmark event

Beck A, Reichert JM. MAbs. 5:621-3 (2013)

Quality considerations draft guidance

Focuses on analytical studies that may be relevant to assessing the similarity between a proposed biosimilar protein product and a reference product. General principles:

Importance of extensive analytical physico-chemical and biological characterisation. Identification of lots used in the various analyses for biosimilarity determination https://www.ibbr.umd.edu/sites/default/files/public_page/Tarlov%20-%20Biomanufacturing%20Summit.pdf

Utility of a NIST mAb Reference Material

Used to distinguish analytical variability from product variability and cross-check analytical methods

Publically available, certified material with historical characterization data representative of a large class of biotherapeutic

Used to reconcile differences between orthogonal methods measuring same attribute

Used in qualification or assessment of changing analytical test methods

Used to assess performance of new analytical technologies

https://www.ibbr.umd.edu/sites/default/files/public_page/Tarlov%20-%20Biomanufacturing%20Summit.pdf

“Standard Reference Material” – distributed from NIST

frozen bulk (100 mg/mL, ≥ 98% purity) supplied in vials at 10 mg/mL & 100mg/ml (1 mL per vial) in 12.5 mM L-Histidine/12.5 mM L-Histidine HCl monohydrate (pH 6.0). Material is NOT FOR HUMAN OR ANIMAL USE

Humanized IgG1.G1m(3);Km3 expressed in NSO cells

Establishment of a “Standard Reference Material” – under the auspices of NIST and ACS

Participating laboratories

Primary Structure •  Intact Mass- intact, reduced, partial digests (Fab, Fc ); • Peptide Map- native and RA; microwave, pressure; column phases

(RP, HILIC, PGC) Sequence Variant analysis

Post-translational and chemical mechanisms • Glycoanalysis ; 2-AB labeling, permethylated MALDI, and

exoglycosidase analysis of glycan composition and structure; LC-MS peptide mapping

Higher Order Structural • NMR; X-ray Crystallography; HDX; Oxidative Footprinting; Ion Mobility; Covalent crosslinking

Orthogonal techniques and assays employed

Biophysical § CD- AUC- ITC/DSC- FT-IR Analysis of the Fab, Fc, and intact mAb; DSC, DLS, membrane confined capillary electrophoresis

Separations

• cIEF; cSDS; SEC; IEX; HIC; HIC; SEC HPLC; CEX

Functional • Biacore; Elisa; Celisa; FcRn; HCP

Coefficient of absorption • AAA/OD280/ extinction coefficient

Orthogonal techniques and assays employed

“Current, State of the Art and Emerging Technologies for the Analysis of Monoclonal Antibodies”.

Editors:

John Schiel, Bioanalytical Science Group, NIST

Darryl Davis, Principal Scientist, Janssen

Oleg Borisov, Principal Scientist, Amgen

Comparability

Volume 1: Monoclonal Antibody Therapeutics: Structure, Function, and Regulatory Space

Chapter 2: Monoclonal antibodies: mechanisms of action – Roy Jefferis

Volume 2: Biopharmaceutical Characterization: The NIST mAb Case Study

Volume 3: Defining the Next Generation of Analytical and Biophysical Techniques

Jefferis R. Arch. Biochem. Biophys. 526:159-662 (2012)

The F1000Research Antibody Validation

Well validated antibodies are crucial to progress in a wide range of life science disciplines. Validating an antibody is a complex and ongoing process.

Helsby MA. et al. F1000Research 3:241 (2014)

Reproducibility: Standardize antibodies used in research

Bradbury A. & Pluckthun A. Nature 518:27-29 (2015)

Present and future challenges – innovative products based on IgG1/2/3/4

Next generation biobetter anti-CD20 antibodies

Desjarlais JR. Lazar G. Exp Cell Res. 3171278-85. (2011)

Obinutuzumab

Ublituximab

Antibody-drug conjugates (ADC)

Float like a butterfly, sting like a bee

Woof JM et al., Mucosal Immunol. 4:590-7 (2011)

Recombinant IgA antibodies are in the pipeline

Induction of tolerance or immunosuppression:

an active mechanism of self/non-self discrimination

Gonzalez S et al. Self/Nonself 2:19-25 (2011)

Operational tolerance Immunosuppression Induction of antigen-specific tolerance

Langer R. et al., PNAS 112:E156-65 (2015)

Innovation

Peanut allergy breakthrough: Children can build up tolerance

Anagnostou K. et al. Arch Dis Child. 100:68-72. (2015) Anagnostuo K et al. Lancet 383(9925):1297-304 (2014)

Induction of antigen-specific tolerance Repeated low doses of aggregate free protein

Jefferis R. Isotype & glycoform selection for antibody therapeutics. Arch Biochem Biophys 526:159-166 (2012)

Davies AM., Jefferis R., Sutton BJ. Crystal structure of deglycosylated IgG4-Fc Mol Immunol. 62:46-53 (2014)

Davies AM, Jefferis, Sutton BJ. Structural determinants of unique properties of human IgG4-Fc. J Mol Biol. 426:630-644 (2014)

Jefferis R. Monoclonal Antibodies: Mechanisms of action. In: Current state of the art for the characterisation of mAbs. Eds: D. Davis, J. Schiel & O. Borisov ACS 2015