Therapeutic Fc-Fusion Proteins E-Book

• Herausgeber: Wiley-VCH
• Kategorie: Wissenschaft und neue Technologien
• Sprache: Englisch

Seitenzahl: 738

Veröffentlichungsjahr: 2013

Beschreibung

Edited by three pioneers in the field, each with longstanding experience in the biotech industry, and a skilled scientific writer, this is the first book to cover every step in the development and production of immunoglobulin Fc-fusion proteins as therapeutics for human disease: from choosing the right molecular design, to pre-clinical characterization of the purified product, through to batch optimization and quality control for large-scale cGMP production. The whole of the second part is devoted to case studies of Fc-fusion proteins that are now commercially successful products. In this section, the authors, several of whom were personally involved in clinical development of the products themselves, detail the product?s background and give insight into issues that were faced and how these issues were overcome during clinical development. This section also includes a chapter on promising new developments for the future. An invaluable resource for professionals already working on Fc-fusion proteins and an excellent and thorough introduction for physicians, researchers, and students entering the field.

Leseprobe

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Contents

Cover

Related Titles

Title Page

Copyright

Preface

List of Contributors

Chapter 1: Introduction: Antibody Structure and Function

1.1 Introduction to Antibodies

1.2 General Domain and Structure of IgG

1.3 The Neonatal Fc Receptor

1.4 Introduction to FcγR- and Complement-Mediated Effector Functions

1.5 Current Trends in Antibody Engineering

References

Part One: Methods of Production for Fc-Fusion Proteins

Chapter 2: Fc-Fusion Protein Expression Technology

2.1 Introduction

2.2 Expression Systems Used for Fc-Fusion Proteins

2.3 Summary

References

Chapter 3: Cell Culture-Based Production

3.1 Introduction

3.2 Basic Aspects of Industrial Cell Culture

3.3 Specific Process Considerations for Fc-Fusion Molecules

3.4 Case Studies

3.5 Conclusions

References

Chapter 4: Downstream Processing of Fc-Fusion Proteins

4.1 Introduction and Overview of Fc-Fusion Proteins

4.2 Biochemistry of Fc-Fusion Proteins

4.3 Purification of Fc-Fusion Proteins from Mammalian Cells

4.4 Purification of Fc-Fusion Protein from Microbial Systems

4.5 Future Innovations in Fc-Fusion Protein Downstream Processing

4.6 Conclusions

References

Chapter 5: Formulation, Drug Product, and Delivery: Considerations for Fc-Fusion Proteins

5.1 Challenges of Molecule Design and Protein Formulation

5.2 The Promise of Fc-Fusion Proteins

5.3 Current Landscape of Commercial Antibody-Related Products

5.4 Fc Conjugates Compared to mAb Counterparts

5.5 Factors in Selecting Liquid versus Lyophilized Formulations

5.6 Advantages and Disadvantages of a Lyophilized Product

5.7 The General Lyophilization Formulation Strategy for Fc-Fusion Proteins

5.8 Bulking Agent

5.9 Surfactant

5.10 The Impact of Residual Moisture

5.11 Practical Considerations for Low-Protein-Concentration Lyophilized Products

5.12 Drug Delivery Considerations

5.13 Device Considerations

5.14 Assessing Feasibility of a Multidose Formulation

5.15 Overage Considerations

5.16 Summary

References

Chapter 6: Quality by Design Applied to a Fc-Fusion Protein: A Case Study

6.1 Introduction

6.2 Critical Quality Attributes

6.3 Critical Process Parameters

6.4 Process Characterization

6.5 Global Multistep Design Space

6.6 Robustness Studies

6.7 Adaptive Strategy

6.8 Engineering Design Space

6.9 Control Strategy

6.10 Continuous Process Verification

6.11 Expanded Change Protocol and Continual Improvement

6.12 Business Case

References

Chapter 7: Analytical Methods Used to Characterize Fc-Fusion Proteins

7.1 Background

7.2 Product Characterization

7.3 Characterization of the Reference Standard

7.4 Typical Product Release and Stability Assays

7.5 Analytical Method Qualification and Validation

References

Part Two: Case Studies of Therapeutic Fc-Fusion Proteins

Chapter 8: Introduction to Therapeutic Fc-Fusion Proteins

8.1 Therapeutic Fc-Fusion Proteins

8.2 Background

8.3 Fc-Fusion Constructs Have Increased In Vivo Stability

8.4 Immunoglobulin-Mediated Effector Function

8.5 Considerations in Fc-Fusion Protein Design

8.6 Fc-Fusion Proteins Approved for Use in the United States

8.7 Concluding Remarks

References

Chapter 9: Alefacept

9.1 Introduction

9.2 Chronic Plaque Psoriasis

9.3 Conventional Treatments for Psoriasis

9.4 Preclinical Development

9.5 Preclinical Primate Studies

9.6 Phase 1 and 2 Human Clinical Studies

9.7 Phase 3 Studies

9.8 Clinical Pharmacology

9.9 Clinical Safety

9.10 Amevive Discontinued

References

Chapter 10: Etanercept

10.1 Introduction

10.2 Design, Construction, and Characterization of TNFR-Fc-Fusion Protein

10.3 Etanercept Preclinical Development

10.4 Etanercept Key Clinical Trials

10.5 Competitive Landscape

10.6 Conclusions

References

Chapter 11: Abatacept and Belatacept

11.1 Introduction

11.2 Design, Construction, and Characterization of Abatacept

11.3 Immunosuppressive Properties of Abatacept

11.4 Rational Design and Characterization of Belatacept

11.5 Belatacept Activity in Primate Renal Transplant Studies

11.6 Abatacept Clinical Development

11.7 Belatacept Clinical Development

11.8 Concluding Remarks

References

Chapter 12: Aflibercept

12.1 Introduction

12.2 Clinical Indications

12.3 Characterization of Aflibercept

12.4 Preclinical Studies with Aflibercept

12.5 Clinical Studies with Aflibercept

12.6 Summary

References

Chapter 13: Recombinant Factor VIII– and Factor IX–Fc Fusions

13.1 Introduction

13.2 Structure and Function of Factor IX and Factor VIII

13.3 Rationale and Design of rFIXFc- and rFVIIIFc-Fusion Proteins

13.4 Development of a Clinical Candidate and Beyond

References

Index

Limit of Liability/Disclaimer of Warranty: While the publisher and author have used their best efforts in preparing this book, they make no representations or warranties with respect to the accuracy or completeness of the contents of this book and specifically disclaim any implied warranties of merchantability or fitness for a particular purpose. No warranty can be created or extended by sales representatives or written sales materials. The Advice and strategies contained herein may not be suitable for your situation. You should consult with a professional where appropriate. Neither the publisher nor authors shall be liable for any loss of profit or any other commercial damages, including but not limited to special, incidental, consequential, or other damages.

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© 2014 Wiley-VCH Verlag GmbH & Co. KGaA, Boschstr. 12, 69469 Weinheim, Germany

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All rights reserved (including those of translation into other languages). No part of this book may be reproduced in any form – by photoprinting, microfilm, or any other means – nor transmitted or translated into a machine language without written permission from the publishers. Registered names, trademarks, etc. used in this book, even when not specifically marked as such, are not to be considered unprotected by law.

Print ISBN: 978-3-527-33317-2

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Cover Design Adam-Design, Weinheim, Germany

Preface

Fc-fusion proteins – engineered polypeptides that combine biologically active peptides or protein domains with the crystallizable fragment (Fc) domain of an antibody – have become widely used agents both in research and in clinical practice. The fact that these molecules resemble antibodies in so many aspects of structure, function, expression, purification, and pharmacology has enabled them to be rapidly integrated into a variety of assays, preclinical studies, and clinical applications through leveraging the prior experience with monoclonal antibodies. In the years following the 1989 report from Genentech by Dan Capon and colleagues on an Fc-fusion protein or “immunoadhesin” composed of CD4 linked to an antibody Fc, a variety of different receptor extracellular domains were produced in this format. An earlier volume, Antibody Fusion Proteins, by Chamow and Ashkenazi (Wiley, 1999) highlighted progress up to the stage of the first therapeutic Fc fusions progressing through clinical trials. Etanercept became the first FDA-approved therapeutic fusion protein in 1998 and has since become one of the most clinically and commercially successful therapeutics. However, the story of therapeutic Fc fusions does not end here. On the contrary, a growing number of these molecules are being developed as biotherapeutics, including Fc-fusion proteins composed of heterodimeric polypeptide chains and others containing novel peptide mimotopes attached to Fc fragments. We therefore thought it important to review the literature and experience in developing this novel class of biologics – hence the current volume, Therapeutic Fc-Fusion Proteins, which brings up-to-date information on the processes of designing and producing these molecules and highlights some of the most prominent case studies from clinical experience.

Owing to the crucial components of antibody structure and function in the design, production, and use of therapeutic Fc fusions, we begin the book with an extensive introduction to the structure and function of IgG molecules (Chapter 1). This is followed by Part One, a series of chapters summarizing state-of-the-art approaches for producing therapeutic Fc proteins: Chapter 2 presents the principles of design and expression systems; Chapter 3, cell culture production; Chapter 4, downstream processing; Chapter 5, formulation and delivery; Chapter 6, quality by design; and Chapter 7, analytical characterization. These chapters provide a roadmap for the development and life cycle of manufacturing processes for therapeutic Fc fusions. Part Two begins with a synopsis (Chapter 8) of clinically significant Fc-fusion proteins that have been approved or are in late-stage clinical trials. Subsequent chapters present case studies of a subset of these, selected for their unique features in terms of molecular design and/or mechanism of action: alefacept, a lymphocyte function-associated antigen 3 (LFA-3) fusion (Chapter 9); etanercept, a tumor necrosis factor (TNF) receptor fusion (Chapter 10); abatacept and belatacept, cytotoxic T-lymphocyte antigen 4 (CTLA-4) fusions (Chapter 11); aflibercept, a vascular endothelial growth factor (VEGF) receptor fusion (Chapter 12); and factor VIII/IX fusions (Chapter 13). In several cases, we have included authors who were involved directly in development of the Fc-fusion protein products about which they have written. We believe that these accounts of the biologics development process in the context of a range of biological mechanisms and disease indications provide important lessons for the development of future therapeutic Fc-fusion proteins.

We thank all of the contributors to this book for taking the time to write what we hope you will find are useful discussions of these topics. We also thank Laura Shih, Wendy Lin, and Anne Chassin du Guerny and the editorial staff of Wiley-Blackwell for their editing support.

San Mateo, CASteven M. ChamowEl Granada, CAHenry LowmanLexington, MAThomas RyllSanta Fe, NMDeborah Farson

November 2013

List of Contributors

Judy R. Berlfein

Biogen Idec

Hemophilia Research

14 Cambridge Center

Cambridge, Massachusetts 02142

USA

Jody D. Berry

BD Biosciences

Antibody Discovery

10770 North Torrey Pines Road

La Jolla, California 92037

USA

Hervé Broly

Merck Serono SA – Corsier sur Vevey

Department of Biotech Process Sciences

Zone Industrielle B

1809 Fenil sur Corsier

Switzerland

Wenjin Cao

Amgen, Inc.

Drug Product Development

1 Amgen Center Drive

Thousand Oaks, California 91320

USA

Steven M. Chamow

Chamow & Associates, Inc.

San Mateo, California 94403

USA

Javier Chaparro-Riggers

Rinat-Pfizer Inc.

Protein Engineering Department

230 E. Grand Avenue

South San Francisco, California 94080

USA

Alex Eon-Duval

Merck Serono SA – Corsier sur Vevey

Department of Biotech Process Sciences

Zone Industrielle B

1809 Fenil sur Corsier

Switzerland

Deborah A. Farson

FarsonInk

Santa Fe, New Mexico 87505

USA

Janean Fisher

BD Biosciences

Antibody Discovery

10770 North Torrey Pines Road

La Jolla, California 92037

USA

Ralf Gleixner

F. Hoffmann-La Roche Ltd

Grenzacherstr. 124

4070 Basel

Switzerland

Uwe Gottschalk

Sartorius-Stedim Biotech

August-Spindler-Str. 11

37079 Goettingen

Germany

Johanna Grossman

San Francisco, California 94123

USA

Yao-Ming Huang

Biogen Idec

BioProcess Development

5000 Davis Drive Research Triangle Park, NC 27709

USA

Esohe Idusogie

OncoMed Pharmaceuticals

800 Chesapeake Drive

Redwood City, California 94063

USA

Rashmi Kshirsagar

Biogen Idec

BioProcess Development

14 Cambridge Center

Cambridge, Massachusetts 02142

USA

Angela L. Linderholm

Davis, California 95616

USA

Ella Mendoza

BD Biosciences

Antibody Discovery

10770 North Torrey Pines Road

La Jolla, California 92037

USA

Massimo Morbidelli

Institute for Chemical and Bioengineering

Department of Chemistry and Applied Biosciences

ETH Zurich

Wolfgang-Pauli-Strasse 10

8093 Zurich

Switzerland

Michael Mulkerrin

OncoMed Pharmaceuticals

800 Chesapeake Drive

Redwood City, California 94063

USA

Benjamin Neunstoecklin

Institute for Chemical and Bioengineering

Department of Chemistry and Applied Biosciences

ETH Zurich

Wolfgang-Pauli-Strasse 10

8093 Zurich

Switzerland

Robert J. Peach

Receptos, Inc.

10835 Road to the Cure, Suite #205

San Diego, California 92121

USA

Robert T. Peters

Biogen Idec

Hemophilia Research

14 Cambridge Center

Cambridge, Massachusetts 02142

USA

Deirdre Murphy Piedmonte

Amgen, Inc.

Drug Product Development

1 Amgen Center Drive

Thousand Oaks, California 91320

USA

Jaume Pons

Rinat-Pfizer Inc.

Protein Engineering Department

230 E. Grand Avenue

South San Francisco, California 94080

USA

Arvind Rajpal

Rinat-Pfizer Inc.

Protein Engineering Department

230 E. Grand Avenue

South San Francisco, California 94080

USA

Margaret Speed Ricci

Amgen, Inc.

Drug Product Development

1 Amgen Center Drive

Thousand Oaks, California 91320

USA

Thomas Ryll

Biogen Idec

BioProcess Development

14 Cambridge Center

Cambridge, Massachusetts 02142

USA

Abhinav A. Shukla

KBI Biopharma

1101 Hamlin Road

Durham, North Carolina 27704

USA

Miroslav Soos

Institute for Chemical and Bioengineering

Department of Chemistry and Applied Biosciences

ETH Zurich

Wolfgang-Pauli-Strasse 10

8093 Zurich

Switzerland

Pavel Strop

Rinat-Pfizer Inc.

Protein Engineering Department

230 E. Grand Avenue

South San Francisco, California 94080

USA

Dwayne Stupack

University of California

Department of Reproductive Medicine

San Diego, California 92093

USA

Pascal Valax

Merck Biodevelopment Site Montesquieu 1 Rue Jacques Monod 33650 Martillac France

Barbara Woppmann

Biogen Idec

BioProcess Development

14 Cambridge Center

Cambridge, Massachusetts 02142

USA

Catherine Yang

BD Biosciences

Antibody Discovery

10770 North Torrey Pines Road

La Jolla, California 92037

USA

Ping Y. Yeh

Amgen, Inc.

Drug Product Development

1 Amgen Center Drive

Thousand Oaks, California 91320

USA

Yik Andy Yeung

Rinat-Pfizer Inc.

Protein Engineering Department

230 E. Grand Avenue

South San Francisco, California 94080

USA

Shanique Young

University of California

Department of Reproductive Medicine

San Diego, California 92093

USA

1

Introduction: Antibody Structure and Function

Arvind Rajpal, Pavel Strop, Yik Andy Yeung, Javier Chaparro-Riggers, and Jaume Pons

1.1 Introduction to Antibodies

Antibodies, a central part of humoral immunity, have increasingly become a dominant class of biotherapeutics in clinical development and are approved for use in patients. As with any successful endeavor, the history of monoclonal antibody therapeutics benefited from the pioneering work of many, such as Paul Ehrlich who in the late nineteenth century demonstrated that serum components had the ability to protect the host by “passive vaccination” [1], the seminal invention of monoclonal antibody generation using hybridoma technology by Kohler and Milstein [2], and the advent of recombinant technologies that sought to reduce the murine content in therapeutic antibodies [3].

During the process of generation of humoral immunity, the B-cell receptor (BCR) is formed by recombination between variable (V), diversity (D), and joining (J) exons, which define the antigen recognition element. This is combined with an immunoglobulin (Ig) constant domain element (μ for IgM, δ for IgD, γ for IgG (gamma immunoglobulin), α for IgA, and for IgE) that defines the isotype of the molecule. Sequences for these V, D, J, and constant domain genes for disparate organisms can be found through the International ImMunoGeneTics Information System® [4]. The different Ig subtypes are presented at different points during B-cell maturation. For instance, all naïve B cells express IgM and IgD, with IgM being the first secreted molecule. As the B cells mature and undergo class switching, a majority of them secrete either IgG or IgA, which are the most abundant class of Ig in plasma.

Characteristics like high neutralizing and recruitment of effector mechanisms, high affinity, and long resident half-life in plasma make the IgG isotype an ideal candidate for generation of therapeutic antibodies. Within the IgG isotype, there are four subtypes (IgG1–IgG4) with differing properties (Table 1.1). Most of the currently marketed IgGs are of the subtype IgG1 (Table 1.2).

Table 1.1 Subtype properties.

Table 1.2 Marketed antibodies and antibody derivatives by target.

The ability of antibodies to recognize their antigens with exquisite specificity and high affinity makes them an attractive class of molecules to bind extracellular targets and generate a desired pharmacological effect. Antibodies also benefit from their ability to harness an active salvage pathway, mediated by the neonatal Fc receptor (FcRn), thereby enhancing their pharmacokinetic (PK) life span and mitigating the need for frequent dosing. The antibodies and antibody derivatives approved in the United States and the European Union (Table 1.2) span a wide range of therapeutic areas, including oncology, autoimmunity, ophthalmology, and transplant rejection. They also harness disparate modes of action like blockade of ligand binding and subsequent signaling, and receptor and signal activation, which target effector functions (antibody-dependent cellular cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC)), and delivery of cytotoxic payload.

Antibodies are generated by the assembly of two heavy chains and two light chains to produce two antigen-binding sites and a single constant domain region (Figure 1.1, panel a). The constant domain sequence in the heavy chain designates the subtype (Table 1.1). The light chains can belong to two families (λ and κ), with most of the currently marketed antibodies belonging to the κ family.

Figure 1.1 Structure and features of the IgG and its interactions. (a) The structure of a full-length IgG is shown in ribbon representation with transparent molecular surface. One heavy chain is shown in blue and one light chain in magenta. The other heavy chain and light chain are shown in gray for clarity. In this orientation, two Fab domains sit on top of the Fc domain and are connected in the middle by the hinge region. The Fab domain is composed of the heavy chain VH and CH1 domains and the light chain VL and CL domains–Protein Data Bank (PDB) [5] code 1HZH [6]. (b) Each variable domain contains three variable loops (L1–L3 on light chain and H1–H3 on heavy chain) that make up the antigen-binding site–PDB code 1HZH [6]. (c) The Fc region is composed of the dimer of CH2 and CH3 domains. The CH3 domains form a tight interaction while the CH2 domains interact through protein–protein, protein–carbohydrate, and carbohydrate–carbohydrate contacts–PDB code 1HZH [6]. (d) The hinge region is composed of a flexible region covalently tied together through disulfide bridges. Structures of the FcγRIIIa and FcγRIIa bound to the Fc are shown. The structures reveal that both receptors bind to the CH2 domain near the hinge and carbohydrates and upon their binding create an asymmetry such that the second FcγR is unable to bind. In this panel, FcγRIII is shown in green, and the FcγRII is shown in purple–PDB codes 3RY6 [7] and 1T83 [8]. (e) The crystal structure of the complex between the Fc and FcRn reveals that FcRn binds between the C2 and C3 domains in the Fc. FcRn chains are shown in red and orange–PDB code 1FRT [9]. (f) Interestingly, the same region also binds to bacterial Protein A commonly used for purification–PDB code 1FC2 [10].