Lipospheres in

Drug Targets and Delivery

Approaches, Methods,
and Applications

© 2005 by CRC Press LLC

 Lipospheres in
Drug Targets and Delivery
Approaches, Methods,
and Applications

EDITED BY

Claudio Nastruzzi

CRC PR E S S
Boca Raton London New York Washington, D.C.
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Library of Congress Cataloging-in-Publication Data

Lipospheres in drug targets and delivery : approaches, methods, and

applications / edited by Claudio Nastruzzi.
p. ; cm.
Includes bibliographical references and index.
ISBN 0-8493-1692-8 (alk. paper)
1. Liposomes—Therapeutic use. 2. Microspheres (Pharmacy). 3.
Nanoparticles. 4. Drug carriers (Pharmacy). 5. Drug targeting.
[DNLM: 1. Liposomes—therapeutic use. 2. Drug Delivery
Systems—methods. 3. Lipids—therapeutic use. QU 93 L7658 2004] I.
Nastruzzi, Claudio. II. Title.

RS201.L55L595 2004
615′.7--dc22 2004051936

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Preface
Colloidal drug carriers, such as liposomes and lipid nanoparticles, are able to modify
the in vivo distribution of associated substances. They, therefore, can be used to
improve the therapeutic index of drugs by increasing their efficacy or reducing their
toxicity. If these delivery systems are carefully designed with respect to the target
and route of administration, they help us to overcome some of the delivery problems
posed by new classes of active molecules, such as peptides, proteins, genes, and
oligonucleotides. They may also extend the therapeutic potential of established drugs,
such as doxorubicin and amphotericin B.
This book describes the use of lipid-based nano- and microparticulate carriers
in such applications. It presents innovative methods of delivering active biochemicals
to different systems, discusses lipospheres as a technical solution to problems asso-
ciated with controlled release of biochemicals, covers lipospheres as carriers for
vaccines, and finally provides procedures for specific applications and describes
biological systems.
With the identification, characterization, and cloning of specific growth factors,
recombinant proteins are now widely used in the clinic. The use of recombinant
hematopoietic growth factors has, for example, allowed the clinical manipulation of
the hematopoietic system. Recombinant human granulocyte colony-stimulating fac-
tor and granulocyte-macrophage colony-stimulating factor are now widely used to
mobilize hematopoietic stem cells, thereby providing a source of hematopoietic stem
cells for autologous or allogeneic transplantation.
However, one disadvantage associated with the use of most recombinant molecules
is their rapid clearance. Because of this rapid clearance, recombinant molecules
require repeated administration to achieve biological efficacy. Initially, continuous
infusion was used to address this pharmacological deficiency. Continuous infusion
has the advantage of delivering drugs in a controlled manner and is particularly
appropriate when it is important to maintain constant plasma drug concentrations.
However, the requirement for continuous venous access and the use of ambulatory
pumps limits its use.
In this context, other approaches have been developed to improve the pharmaco-
kinetic and pharmacodynamic properties of recombinant proteins in vivo. These have
included the addition of polyethylene glycol to the recombinant molecules (PEGyla-
tion) and the use of sustained-release delivery systems. One goal of these approaches
is to achieve clinical efficacy and lower the number of administrations, possibly to
single injections, and thereby increase patient compliance. In addition to improving
the pharmacokinetic and pharmacodynamic profile of recombinant molecules, sus-
tained release may also increase the biological activity of specific molecules.
Moreover, it should be acknowledged that the use of an efficient carrier for nucleic
acid–based medicines is considered to be a determinant factor for the successful

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application of gene therapy. However, the drawbacks associated with the use of viral
vectors, namely, those related to safety problems, have prompted investigators to
develop alternative methods for gene delivery, with cationic lipid-based systems
being the most representative systems. Despite extensive research in the last decade
on the use of cationic liposomes as gene transfer vectors, and the development of
elegant strategies to enhance their biological activity, these systems are still far from
being viable alternatives to the use of viral vectors in gene therapy.
Finally, in this book considerations are made regarding the structure–activity
relationships of cationic liposphere/DNA complexes, and the key formulations are
presented and discussed in terms of their effect on biological activity.

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Editor

Claudio Nastruzzi was born in Ferrara, Italy, on March 29, 1958. In 1983, he earned
his undergraduate degree in pharmaceutical chemistry at the University of Ferrara, and
in the late 1980s he was a fellow at the university’s Department of Pharmaceutical
Science, working on natural compound synthesis (prostaglandins and leukotrienes)
and the characterization of isosazolic and isosazolinic nuclei reactivity. In 1988, he
obtained his Ph.D. in pharmaceutical science with a dissertation on the synthesis
and antitumor and antimetastatic activity of aromatic polyamidines.
During the late 1990s, Dr. Nastruzzi worked with Professor P. L. Luisi as a
postdoctoral fellow at the Institute for Polymers at the Swiss Federal Institute of
Technology in Zürich. As a postdoctoral fellow at the Department of Pharmaceutical
Sciences at the University of Ferrara, he focused on the production and character-
ization of liposomes specially designed for retinoid delivery, as well as on biophysical
studies and activity of in vitro cultured cell lines. In 1991, he obtained a researcher
position in this department, where he devoted his energy to the production of
microspheres, liposomes, and microemulsions for the controlled delivery of
biological response modifiers.
Since 1998, Dr. Nastruzzi has been an associate professor in the Department of
Chemistry and Pharmaceutical Technology at the University of Perugia, Italy. His
main topics of interest include the production and characterization of innovative

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dermatologic and cosmetic formulations for dermal and transdermal delivery (phos-
pholipid-based microemulsion gels, cubic phases, liposomes, and niosomes); the
production and characterization of microspheres and solid lipid microparticles; and
liposome-based formulations for gene delivery.
Dr. Nastruzzi has published more than 90 papers in international journals and
has presented more than 80 contributions to national and international congresses.

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Contributors
Heike Bunjes Karsten Mäder
Department of Pharmaceutical Institute of Pharmaceutics and
Technology Biopharmacy
Friedrich Schiller University of Jena Martin Luther University of Halle
Jena, Germany Halle, Germany

Rita Cortesi Wolfgang Mehnert

Department of Pharmaceutical Sciences Department of Pharmaceutical
University of Ferrara Technology
Ferrara, Italy Free University of Berlin
Abraham J. Domb Berlin, Germany
Department of Medicinal Chemistry
and Natural Products Boaz Mizrahi
Hebrew University of Jerusalem Department of Medicinal Chemistry
Jerusalem, Israel and Natural Products
Hebrew University of Jerusalem
Elisabetta Esposito Jerusalem, Israel
Department of Pharmaceutical Sciences
University of Ferrara R.H. Müller
Ferrara, Italy Department of Pharmaceutics,
Biopharmaceutics, and Biotechnology
Aviva Ezra Free University of Berlin
Department of Medicinal Chemistry Berlin, Germany
and Natural Products
Hebrew University of Jerusalem Carsten Olbrich
Jerusalem, Israel Schering AG
Berlin, Germany
Achim Göpferich
Department of Pharmaceutical
Technology Monika Schäfer-Korting
University of Regensburg Department of Pharmacology and
Regensburg, Germany Toxicology
Free University of Berlin
Stephanie Könnings Berlin, Germany
Department of Pharmaceutical
Technology Nadja Schöler
University of Regensburg Schering AG
Regensburg, Germany Berlin, Germany

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Kerstin Tabatt Sylvia A. Wissing

PenCef Pharma GmbH Drug Delivery Services GmbH
Berlin, Germany Kronshagen, Germany

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Table of Contents
Chapter 1
Solid Lipid Nanoparticles — Concepts, Procedures, and Physicochemical
Aspects.......................................................................................................................1
Karsten Mäder and Wolfgang Mehnert

Chapter 2
Production of Lipospheres for Bioactive Compound Delivery ..............................23
Elisabetta Esposito, Rita Cortesi, and Claudio Nastruzzi

Chapter 3
Characterization of Solid Lipid Nano- and Microparticles ....................................41
Heike Bunjes

Chapter 4
Lipospheres as Delivery Systems for Peptides and Proteins..................................67
Stephanie Könnings and Achim Göpferich

Chapter 5
Lipospheres for Vaccine Delivery ...........................................................................87
Abraham J. Domb, Aviva Ezra, and Boaz Mizrahi

Chapter 6
Solid Lipid Nanoparticles: Interaction with Cells, Cytokine Production, and
Enzymatic Degradation .........................................................................................101
Carsten Olbrich, Kerstin Tabatt, Sylvia A. Wissing, Nadja Schöler, and
R.H. Müller

Chapter 7
Delivery of Lipophilic Compounds with Lipid Nanoparticles — Applications
in Dermatics and for Transdermal Therapy ..........................................................127
Monika Schäfer-Korting and Wolfgang Mehnert

Chapter 8
Cationic Lipospheres as Delivery Systems for Nucleic Acid Molecules.............143
Rita Cortesi, Elisabetta Esposito, and Claudio Nastruzzi

© 2005 by CRC Press LLC