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Beschreibung

Here, front-line researchers in the booming field of nanobiotechnology describe the most promising approaches for bioinspired drug delivery, encompassing small molecule delivery, delivery of therapeutic proteins and gene delivery. The carriers surveyed include polymeric, proteinaceous and lipid systems on the nanoscale, with a focus on their adaptability for different cargoes and target tissues. Thanks to the broad coverage of carriers as well as cargoes discussed, every researcher in the field will find valuable information here.

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Table of Contents

Cover

Related Titles

Title Page

Copyright

Foreword

List of Contributors

Preface

Chapter 1: Backbone Degradable and Coiled-Coil Based Macromolecular Therapeutics

1.1 Introduction

1.2 Water-Soluble Polymers as Carriers of Anticancer Drugs

1.3 Drug-Free Macromolecular Therapeutics – A New Paradigm in Drug Delivery

1.4 General Summary and Outlook

Acknowledgments

References

Chapter 2: Dendritic Polymers as Targeting Nanoscale Drug Delivery Systems for Cancer Therapy

2.1 Introduction

2.2 Functional Dendritic Polymers Based Drug Delivery Vehicles for Targeting Tumor Therapy via EPR Effect

2.3 Tumor Targeting Moieties Functionalized Dendritic Drug Delivery Vehicles for Cancer Therapy

2.4 Conclusion

References

Chapter 3: Composite Colloidal Nanosystems for Targeted Delivery and Sensing

3.1 Introduction

3.2 Objective

3.3 Cellular Behavior of the Carrier

3.4 Applications

3.5 Conclusions

Abbreviations

References

Chapter 4: Polymeric Micelles for Cancer-Targeted Drug Delivery

4.1 Introduction

4.2 Micelle Formulations in Clinical Development

4.3 Particle Size of Micelles

4.4 Morphology of Micelles

4.5 Targeting Design of Micelles for Enhanced Accumulation and Cell Internalization

4.6 Functional Designs of Micelles

4.7 Design of Micelles for Gene Delivery

4.8 Challenge and Future Perspective

References

Chapter 5: Biomimetic Polymers for In Vivo Drug Delivery

5.1 Introduction

5.2 Commonly Used Biomimetic Polymers and Their Applications in DDS

5.3 Challenges and Perspectives

References

Chapter 6: Drug Delivery from Protein-Based Nanoparticles

6.1 Introduction

6.2 Preparation of Protein-Based Nanoparticles

6.3 Drug Delivery from Albumin-Based Nanoparticles

6.4 Drug Delivery from Gelatin-Based Nanoparticles

6.5 Drug Delivery from Other Protein-Based Nanoparticles

References

Chapter 7: Polymeric Gene Carriers

7.1 Gene Therapy and Gene Carriers

7.2 Polymeric Gene Carriers

7.3 PEI Grafting Modification Polymeric Gene Carriers

7.4 Low Molecular Weight (LWM) PEI Base Polymeric Gene Carriers

7.5 Targeted Shielding System for Polymeric Gene Carriers

7.6 Conclusion

References

Chapter 8: pH-Sensitive Polymeric Nanoparticles as Carriers for Cancer Therapy and Imaging

8.1 Introduction

8.2 pH-Sensitive Polymers

8.3 pH-Sensitive Polymers as Drug Carriers

8.4 pH-Sensitive Polymers for Bioimaging

8.5 Conclusions

References

Chapter 9: Charge-Reversal Polymers for Biodelivery

9.1 Applications of Cationic Polymers in Biodelivery

9.2 Barriers for Cationic Polymers in

In vitro

and

In vivo

Applications

9.3 Characteristic pH Gradients in Tumor Interstitium and Endo/Lysosomes

9.4 Chemistry of Charge-Reversal Polymers Based on Acid-Labile Amides

9.5 Applications of Charge-Reversal Polymers in Biodelivery Systems

9.6 Perspectives

References

Chapter 10: Phenylboronic Acid-Containing Glucose-Responsive Polymer Materials: Synthesis and Applications in Drug Delivery

10.1 Introduction

10.2 PBA-Containing Polymers Operating Under Physiological Conditions

10.3 Chemically Crosslinked PBA-Based Gels

10.4 Self-Assembled PBA-Based Polymer Micelles

10.5 Self-Assembled PBA-Based Polymersomes

10.6 Perspectives

References

Chapter 11: Extracellular pH-Activated Nanocarriers forEnhanced Drug Delivery to Tumors

11.1 Introduction

11.2 Passive and Active Tumor Targeting

11.3 Targeting the Extracellular pH (pH

e

) in Tumors

11.4 Extracellular pH-Induced Drug Delivery to Tumors

11.5 Ligand Exposure by a Shielding/Deshielding Method

11.6 Surface Charge Reversing Nanoparticles

11.7 Conclusion

References

Chapter 12: Stimulation-Sensitive Drug Delivery Systems

12.1 Introduction

12.2 pH-Sensitive Delivery Systems

12.3 Thermo-Sensitive Delivery Systems

12.4 Biomolecule-Sensitive Delivery Systems

12.5 Other Environmentally Sensitive Nanocarriers

12.6 Outlook

References

Index

EULA

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Guide

Cover

Table of Contents

Preface

Chapter 1: Backbone Degradable and Coiled-Coil Based Macromolecular Therapeutics

List of Illustrations

Figure 1.1

Figure 1.2

Figure 1.3

Figure 1.4

Figure 1.5

Figure 1.6

Figure 1.7

Figure 1.8

Figure 1.9

Figure 1.10

Figure 1.11

Figure 2.1

Figure 2.2

Figure 2.3

Figure 2.4

Figure 2.5

Figure 2.6

Figure 2.7

Figure 2.8

Figure 2.9

Figure 3.1

Figure 3.2

Figure 3.3

Figure 3.4

Figure 3.5

Figure 4.1

Figure 4.2

Figure 4.3

Figure 4.4

Figure 4.5

Figure 4.6

Figure 4.7

Figure 4.8

Figure 4.9

Figure 4.10

Figure 6.1

Figure 6.2

Figure 6.3

Figure 6.4

Figure 6.5

Figure 6.6

Figure 6.7

Figure 6.8

Figure 6.9

Figure 7.1

Figure 7.2

Figure 7.3

Figure 7.4

Figure 7.5

Figure 7.6

Figure 7.7

Figure 7.8

Figure 7.9

Figure 7.10

Figure 8.1

Figure 8.2

Figure 8.3

Figure 8.4

Figure 8.5

Figure 8.6

Figure 9.1

Figure 9.2

Figure 9.3

Figure 9.4

Figure 9.5

Figure 9.6

Figure 9.7

Figure 9.8

Figure 10.1

Figure 10.2

Figure 10.3

Figure 10.4

Figure 10.5

Figure 10.6

Figure 10.7

Figure 10.8

Figure 10.9

Figure 10.10

Figure 10.11

Figure 10.12

Figure 10.13

Figure 10.14

Figure 10.15

Figure 10.16

Figure 10.17

Figure 10.18

Figure 10.19

Figure 10.20

Figure 10.21

Figure 10.22

Figure 10.23

Figure 10.24

Figure 10.25

Figure 10.26

Figure 10.27

Figure 10.28

Figure 10.29

Figure 10.30

Figure 10.31

Figure 11.1

Figure 11.2

Figure 11.3

Figure 11.4

Figure 11.5

Figure 11.6

Figure 11.7

Figure 11.8

Figure 11.9

Figure 11.10

Figure 11.11

Figure 11.12

Figure 11.13

Figure 11.14

Figure 11.15

Figure 11.16

Figure 11.17

Figure 11.18

Figure 11.19

Figure 12.1

Figure 12.2

Figure 12.3

List of Tables

Table 5.1

Table 5.2

Table 5.3

Table 5.4

Table 12.1

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Edited by Zhongwei Gu

Bioinspired and Biomimetic Polymer Systems for Drug and Gene Delivery

Editor

Prof. Zhongwei Gu

National Engineering Research Center

for Biomaterials Sichuan Univ.

Wangjiang Road 29

610064 Chengdu

China

All books published by Wiley-VCH are carefully produced. Nevertheless, authors, editors, and publisher do not warrant the information contained in these books, including this book, to be free of errors. Readers are advised to keep in mind that statements, data, illustrations, procedural details or other items may inadvertently be inaccurate.

Library of Congress Card No.: applied for

British Library Cataloguing-in-Publication Data

A catalogue record for this book is available from the British Library.

Bibliographic information published by the Deutsche Nationalbibliothek

The Deutsche Nationalbibliothek lists this publication in the Deutsche Nationalbibliografie; detailed bibliographic data are available on the Internet at <http://dnb.d-nb.de>.

© 2015 Wiley-VCH Verlag GmbH & Co. KGaA, Boschstr. 12, 69469 Weinheim, Germany

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: 978-3-527-33420-9

ePDF ISBN: 978-3-527-67273-8

ePub ISBN: 978-3-527-67272-1

Mobi ISBN: 978-3-527-67271-4

oBook ISBN: 978-3-527-67275-2

Foreword

In recent years, the rapid development of polymer science and advances in modern medicine, pharmacy, biology, and engineering have fostered the emergence of a new field focused on the theory and technology underlying drug delivery. This inter-disciplinary field is called drug delivery systems (DDS). It shows great promise and has become a hotspot in biomedical material research, especially in biomedical polymers.

The successful development of advanced, efficient DDS depends on the design and construction of the materials and micro devices involved. The research frontier focuses mainly on targeted delivery, especially cell and molecular targeting, and on controlled release stimulated by the tissue or cellular microenvironment. The complex in vivo physiological and pathological environment often obscures the effects of active targeting. In this way, producing a highly efficient system capable of active targeting in vivo is the key to improving the efficacy of DDS. Drug release systems capable of biological sensing are called bioinspired and biomimetic delivery systems. They automatically adjust the drug release in response to external stimuli, such as changes in temperature, pH, magnetic fields, ultrasound, and electric fields. They have received a considerable amount of attention from researchers and pharmaceutical companies worldwide. Drug release systems that can be switched on and off via self-feedback upon changes in the chemical or physical signals given off by a lesion or intelligent carrier have drawn particular interest. Systems that can undergo rapid stimuli-responsive controlled release under in vivo microenvironment conditions would be far more useful to actual clinical treatment regimens.

This book embodies the wisdom and achievements of renowned experts and research teams in this field from China, the United States, Germany, Japan, and Korea. The discussion provided herein covers the most important, active, and cutting-edge parts of this field, reflecting the latest developments and trends in DDS research. The chief editor, Professor Zhongwei Gu, studied under the pioneer biomedical polymers in China–Professor Xin-De Feng (Academician of Chinese Academy of Sciences). Gu entered this field in the 1970s and has become a well-known professor of polymer biomaterials in China. It is our hope that this book will promote scientific research and biomedical applications in the vibrant and exciting area. Young academics and professionals interested in DDS may also benefit from this treatise.

We would like to thank all our editors for their hard work and dedication. We would also like to thank John Wiley & Sons Publishing Company and Chemical Industry Press for their forward-looking strategic vision and the timely publication of this book.

August 2014

Professor Ren-Xi Zhuo

Academician of Chinese Academy of Sciences

IUS-BSE Fellow

Wuhan University

Wuhan, China

List of Contributors

Huabing Chen

Soochow University

College of Pharmaceutical Science

Suzhou 215123

China

Xuesi Chen

Chinese Academy of Sciences

Changchun Institute of Applied Chemistry

Renmin Street

Changchun 130022

China

Du Cheng

Sun Yat-sen University

PCFM Lab of Ministry of Education

School of Chemistry and Chemical Engineering

Xingangxi Road 135

Guangzhou 510275

China

Dan Ding

Nanjing University

Laboratory of Mesoscopic Chemistry and Department of Polymer Science & Engineering

College of Chemistry & Chemical Engineering

Hankou Road

Nanjing

China

Jin-Zhi Du

School of Life Sciences

University of Science and Technology of China

Hefei

Anhui 230027

China

Guang Hui Gao

Sungkyunkwan University

Theranostic Macromolecular Research Center

School of Chemical Engineering

Republic of Korea

Zhishen Ge

University of Science and Technology of China

CAS Key Laboratory of Soft Matter Chemistry

Department of Polymer Science and Engineering

Hefei 230026

China

Pilar Rivera Gil

Centre Tecnològic de la Química de Catalunya (CTQC) and

Dpt Physical and Inorganic Chemistry Dpt.

Universitat Rovira i Virgili

Marcel.li Domingo

s/n Edif. N5

Tarragona

Spain

Zhongwei Gu

Sichuan University

National Engineering Research Center for Biomaterials

Chengdu 610064

China

Xiuwen Guan

Chinese Academy of Sciences

Changchun Institute of Applied Chemistry

Renmin Street

Changchun 130022

China

Xiqun Jiang

Nanjing University

Laboratory of Mesoscopic Chemistry and Department of Polymer Science & Engineering

College of Chemistry & Chemical Engineering

Hankou Road

Nanjing

China

Kazunori Kataoka

The University of Tokyo

Department of Materials Engineering

Graduate School of Engineering

7-3-1 Hongo

Bunkyo-ku

Tokyo 8656

Japan

Jindich Kopeek

University of Utah

Department of Pharmaceutics and Pharmaceutical Chemistry

S, 2030 E

Salt Lake City

Utah 84112

USA

and

University of Utah

Department of Bioengineering

Salt Lake City

Utah 84112

USA

Ick Chan Kwon

Korea Institute of Science and Technology

Biomedical Research Center

Republic of Korea

Doo Sung Lee

Sungkyunkwan University

Theranostic Macromolecular Research Center

School of Chemical Engineering

Republic of Korea

Yi Li

Sungkyunkwan University

Theranostic Macromolecular Research Center

School of Chemical Engineering

Republic of Korea

Kui Luo

Sichuan University

National Engineering Research Center for Biomaterials

Chengdu 610064

China

Rujiang Ma

Key Laboratory of Functional Polymer Materials

Ministry of Education

and Institute of Polymer Chemistry

Nankai University

Tianjin 300071

China

Cheng-Qiong Mao

School of Life Sciences

University of Science and Technology of China

Hefei

Anhui 230027

China

Moritz Nazarenus

Philipps University of Marburg

Department of Biophotonics

Institute of Physics

Renthof 7

Marburg

Germany

Wolfgang J. Parak

Philipps University of Marburg

Department of Biophotonics

Institute of Physics

Renthof 7

Marburg

Germany

Kinam Park

Purdue University

Departments of Biomedical Engineering and Pharmaceutics

S. Martin Jischke Drive

West Lafayette

Indiana, 47907

USA

Youqing Shen

Zhejiang University

Center for Bionanoengineering

Department of Chemical and Biological Engineering

Zheda ST

Hangzhou 310027

China

Linqi Shi

Key Laboratory of Functional Polymer Materials

Ministry of Education

and Institute of Polymer Chemistry

Nankai University

Tianjin 300071

China

Xintao Shuai

Sun Yat-sen University

PCFM Lab of Ministry of Education

School of Chemistry and Chemical Engineering

Xingangxi Road 135

Guangzhou 510275

China

Jingxing Si

Zhejiang University

Center for Bionanoengineering

Department of Chemical and Biological Engineering

Zheda ST

Hangzhou 310027

China

Meihua Sui

Zhejiang University

Center for Bionanoengineering

Department of Chemical and Biological Engineering

Zheda ST

Hangzhou 310027

China

Huayu Tian

Chinese Academy of Sciences

Changchun Institute of Applied Chemistry

Renmin Street

Changchun 130022

China

Jun Wang

School of Life Sciences

University of Science and Technology of China

Hefei

Anhui 230027

China

Kai Wang

Zhejiang University

Center for Bionanoengineering

Department of Chemical and Biological Engineering

Zheda ST

Hangzhou 310027

China

Wenping Wang

Ningxia Medical University

Department of Pharmaceutics, School of Pharmacy

Shengli Street

Yinchuan

Ningxia, 750004

China

and

Purdue University

Departments of Biomedical Engineering and Pharmaceutics

S. Martin Jischke Drive

West Lafayette

Indiana, 47907

USA

Jiyuan Yang

University of Utah

Department of Pharmaceutics and Pharmaceutical Chemistry

S, 2030 E

Salt Lake City

Utah 84112

USA

Xian-Zhu Yang

School of Life Sciences

University of Science and Technology of China

Hefei

Anhui 230027

China

You-Yong Yuan

School of Life Sciences

University of Science and Technology of China

Hefei

Anhui 230027

China

Bo Zhang

Zhejiang University

Center for Bionanoengineering

Department of Chemical and Biological Engineering

Zheda ST

Hangzhou 310027

China

and

University of Wyoming

Department of Chemical and Petroleum Engineering

E. Univ. Ave., Laramie

WY 82071

USA

Preface

The rapid development of biomedical materials science and engineering and advances in modern medicine, pharmacy, biology, and engineering have made the great promise of advanced drug delivery systems (DDS) increasingly clear. These DDS systems are made of carrier materials and drugs, including peptides, proteins, antigens, and nucleic acid drugs, allowing the controlled release of active agents. The advanced DDS systems would not only be a revolutionary change from the traditional mode of drug delivery but may also facilitate the development of currently infeasible approaches to the treatment of cancer, cardiovascular disease, AIDS, congenital genetic defects, and other diseases. It has also greatly promoted the development of molecular diagnostic medicine, which may facilitate the early diagnosis and exploration of the pathogenesis and development of disease and the resultant pathological and physiological changes. With the continuous development of new materials and new technologies and urgent clinical needs, the production of carrier materials and the drug controlled-release/delivery system has become an important part of the entire pharmaceutical industry, and is growing to be the most promising sunrise industry, bringing tremendous and far-reaching impact on the global pharmaceutical industry. The forces driving its rapid growth are as follows: First, the research and development of carrier materials capable of restoring and improving the body's physiological functions and delivery systems capable of releasing them in a controlled manner are the main direction of contemporary biomedical material research. Second, diseases that are currently difficult to cure may become treatable or easier to detect early. This may involve a significant reduction in health care costs. Third, drug delivery systems capable of controlled release have several advantages over current systems. They tend to last longer, deliver their payload more efficiently in terms of time, and are less toxic to the patient. This also improves the bioavailability of drugs, especially peptide, protein, and nucleic acid drugs. Fourth, these systems are conducive to the development of new drugs. They can reduce costs, shorten development cycles, and provide a quicker return on investment than conventional drug development.

The designs and preparation techniques of carrier materials and micro-systems are the key to the development of the advanced, efficient drug controlled-release/delivery system and the theranostic micro devices. The cutting-edge areas and future trends of this field mainly include the following aspects.

The first aspect is the intelligent/microenvironment-triggered (stimulus-responsive) DDS. Such systems use the material's ability to detect changes in pH, redox, other chemical signals, temperature, optics, magnetic fields, electrical signals, mechanical signals, other physical signals, and enzymes, receptors, and other biological signals to facilitate responsive drug release. It may be the most promising and the most valuable DDS with the greatest prospect in clinical applications in the t21st century. The key technology in this field is to improve the sensitivity of the material and the DDS, in order to achieve rapid response in vivo.

The second aspect is the targeted DDS itself, which includes passive and active targeting. Passive targeted DDS cannot recognize the target cells, instead relying on the size effect to reach the target site. For example, a drug carrier system that is 20–200 nm in size triggers an enhanced permeability and retention (EPR) effect based on the defects (holes) in the blood vessels of the tumor tissues, thereby causing efficient local accumulation of drugs. Precise control of the particle size, sufficiently narrow size distribution, and increase in the circulation time in vivo are crucial to passive targeting. Active targeted DDS introduces a targeting group capable of recognizing the target tissue, target cells, or even target molecules. The ability to recognize the cells or molecules of a specific tissue or organ is highly relevant to material preparation. The advantages of this type of DDS are its high selectivity and reduced side effects to normal tissues and organs, which would be very valuable in the treatment of common multiple malignancies.

The third aspect is self-regulated DDS. The system must mimic the complex biochemical processes in vivo and release the drug in accordance with the body's needs, rather than at a stable, predetermined speed. The release mechanisms involve the microporous or bulk diffusion of the polymer reservoir or the enzymatic degradation on the surface of the polymer matrix.

The fourth aspect is time control. To achieve constant speed, zero-order release of the drug is the main direction. The chemical structure, composition, and degradation properties of the material and its capacity for drug penetration and diffusion are key to achieving constant release. Transdermal drug delivery avoids the first-pass effect and gastrointestinal damage because it does not require that the drug pass through the liver. This makes it more practical for patients requiring long-term continuous administration. It is also highly efficient and may reduce the rate of side effects. In addition, the source of the drug, usually a patch, can be removed at any time. The core issues in transdermal drug delivery are producing a polymer film capable of drug transmission, which involves the principles of pharmacodynamics and pharmacokinetics.

This book covers bioinspired and polymer nano drug delivery systems. It is published jointly by John Wiley Publishing Company and China Chemical Industry Press. Its goal is to review relevant progress and the future direction of development in order to promote the development of this field, the clinical application of nano drug delivery systems, and improvements in medicine. To this end, internationally renowned experts in this field co-authored the book with different styles and from different perspectives. The book consists of 12 chapters, mainly including backbone-degradable high molecular weight (second-generation) water-soluble polymer-anticancer drug conjugates, and a new paradigm of drug-free macromolecular therapeutics; nano-targeting drug delivery systems based on peptide dendrimers; nano composite colloidal systems with multiple functions including targeting, stimulus-responsive controlled release, and sensing; multifunctional polymeric micelles that can break various physiological barriers for targeted delivery; biomimetic polymer used for in vivo drug delivery; nanoparticle-based proteins such as gelatin, human serum albumin, collagen, silk protein, casein protein, and elastin-like polypeptides; polymer carriers for gene delivery system and their functional modifications; pH-sensitive nano drug delivery systems for targeted cancer therapy and biological imaging; drug/gene nano delivery system capable of charge flipping; nano drug carrier systems using phenylboronic acid-based glucose-responsive polymeric materials and their gels, micelles, and vesicles; nano delivery systems for cancer treatment co-carrying drugs and siRNA and activated by extracellular pH; and pH, heat, and biological molecule or light-sensitive stimuli-responsive DDS. This book covers the design principles, research and development technologies, and application prospects of advanced and efficient polymer nano DDS, and rather comprehensively discusses the outstanding progresses and future trends of the field. This forward-looking, novel treatise on polymer nano DDS is unique and extremely valuable, and I believe that it will be welcomed by readers. In particular, young scholars interested in the field of polymer nano drug delivery systems will learn a great deal from this book, and may be inspired to carry on their own research.

I would like to thank all the editors and contributing authors for their time and expertise. I am especially grateful to Dr. Gang Wu from the China Chemical Industry Press and Dr. Esakki Rahini from the John Wiley Publishing Company for their contributions to the publishing of this book. I thank Professor Bin He, Professor Yao Wu, and everyone else who have helped. I would also like to sincerely thank Professor Renxi Zhuo of the Chinese Academy of Science, a pioneer and esteemed leader in the field of polymer chemistry and drug delivery systems, for writing the preface for this book and for his guidance and support.

Professor Zhongwei Gu

Chief Scientist of Biomaterials, National 973 Program

IUS-BSE Fellow

Sichuan University

Chengdu, China

1Backbone Degradable and Coiled-Coil Based Macromolecular Therapeutics

Jiyuan Yang and Jindich Kopeek

1.1 Introduction

To address the lack of specificity of low molecular weight drugs for malignant cells, the concept of targeted polymer–drug conjugates was developed in the 1970s. The major rationale for the use of water-soluble polymers as carriers of anticancer drugs is based on the mechanism of cell entry [1–3]. Whereas the majority of low molecular weight drugs enter the cell interior by diffusion through the plasma membrane, the entry of macromolecules is restricted to endocytosis [4]. Macromolecules captured by this mechanism are channeled to the lysosomal compartment of the cell. In addition, moieties that complement cell surface receptors or antigens of a subset of cells may be incorporated into the macromolecular structure and render the conjugate biorecognizable [5–9].

There are numerous reviews, which summarize the rationale, design, synthesis, evaluation, and development of macromolecular therapeutics [5–16]. In this chapter, polymeric carriers based on copolymers of N-(2-hydroxypropyl)methacrylamide (HPMA) are used as an example. However, the conclusions can be considered generally applicable to water-soluble carriers with other chemical structures.

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Lesen Sie weiter in der vollständigen Ausgabe!

Lesen Sie weiter in der vollständigen Ausgabe!

Lesen Sie weiter in der vollständigen Ausgabe!

Lesen Sie weiter in der vollständigen Ausgabe!

Lesen Sie weiter in der vollständigen Ausgabe!

Lesen Sie weiter in der vollständigen Ausgabe!

Lesen Sie weiter in der vollständigen Ausgabe!