(Ebook) New Developments in Medicinal Chemistry by

Marta P. Ortega; Irene C. Gil ISBN 9781617285462,

1617285463 Pdf Download

https://ebooknice.com/product/new-developments-in-medicinal-
chemistry-51368846

★★★★★
4.6 out of 5.0 (26 reviews )

Instant PDF Download

ebooknice.com
 (Ebook) New Developments in Medicinal Chemistry by Marta P.
Ortega; Irene C. Gil ISBN 9781617285462, 1617285463 Pdf
Download

EBOOK

Available Formats

■ PDF eBook Study Guide Ebook

EXCLUSIVE 2025 EDUCATIONAL COLLECTION - LIMITED TIME

INSTANT DOWNLOAD VIEW LIBRARY

We have selected some products that you may be interested in
Click the link to download now or visit ebooknice.com
for more options!.

(Ebook) Medicinal Chemistry by D. Sriram, P. Yogeeswari ISBN

9788131731444, 8131731448

https://ebooknice.com/product/medicinal-chemistry-10886056

(Ebook) New developments in analytical chemistry research by

Granger, Breanna ISBN 9781634634274, 9781634634731, 1634634276,
163463473X

https://ebooknice.com/product/new-developments-in-analytical-chemistry-
research-6856822

(Ebook) Recessions: Prospects and Developments : Prospects and

Developments by Nerea M. Pérez; June A. Ortega ISBN
9781614707349, 1614707340

https://ebooknice.com/product/recessions-prospects-and-developments-
prospects-and-developments-51361314

(Ebook) Nanomaterials Chemistry: Recent Developments and New

Directions by C. N. R. Rao, Achim Müller, Anthony K. Cheetham
ISBN 9783527316649, 3527316647

https://ebooknice.com/product/nanomaterials-chemistry-recent-developments-
and-new-directions-1293968
(Ebook) Annual Reports in Medicinal Chemistry by Desai, Manoj C
ISBN 9780128001677, 9780128003725, 9781299875029, 0128001674,
0128003723, 1299875025

https://ebooknice.com/product/annual-reports-in-medicinal-chemistry-5431298

(Ebook) The Pediatric and Perinatal Autopsy Manual with DVD-ROM

by Marta C. Cohen (editor), Irene Scheimberg (editor) ISBN
9781107646070, 1107646073

https://ebooknice.com/product/the-pediatric-and-perinatal-autopsy-manual-
with-dvd-rom-22209568

(Ebook) New Developments in Sickle Cell Disease Research by P D

O’Malley ISBN 9781594547928, 1594547920

https://ebooknice.com/product/new-developments-in-sickle-cell-disease-
research-6670376

(Ebook) New Developments in Quartz Research: Varieties, Crystal

Chemistry and Uses in Technology : Varieties, Crystal Chemistry
and Uses in Technology by Bruno Novak; Pavla Marek ISBN
9781624172663, 1624172660
https://ebooknice.com/product/new-developments-in-quartz-research-
varieties-crystal-chemistry-and-uses-in-technology-varieties-crystal-
chemistry-and-uses-in-technology-51361174

(Ebook) Chemogenomics in Drug Discovery: A Medicinal Chemistry

Perspective (Methods and Principles in Medicinal Chemistry) by
Hugo Kubinyi, Gerhard Muller, Raimund Mannhold, Gerd Folkers
ISBN 9783527309870, 352730987X
https://ebooknice.com/product/chemogenomics-in-drug-discovery-a-medicinal-
chemistry-perspective-methods-and-principles-in-medicinal-chemistry-1803322
Copyright © 2009. Nova Science Publishers, Incorporated. All rights reserved.

New Developments in Medicinal Chemistry, Nova Science Publishers, Incorporated, 2009. ProQuest Ebook Central,
Copyright © 2009. Nova Science Publishers, Incorporated. All rights reserved.

New Developments in Medicinal Chemistry, Nova Science Publishers, Incorporated, 2009. ProQuest Ebook Central,
 NEW DEVELOPMENTS IN
MEDICINAL CHEMISTRY

No part of this digital document may be reproduced, stored in a retrieval system or transmitted in any form or
by any means. The publisher has taken reasonable care in the preparation of this digital document, but makes no
expressed or implied warranty of any kind and assumes no responsibility for any errors or omissions. No
liability is assumed for incidental or consequential damages in connection with or arising out of information
contained herein. This digital document is sold with the clear understanding that the publisher is not engaged in
rendering legal, medical or any other professional services.
Copyright © 2009. Nova Science Publishers, Incorporated. All rights reserved.

New Developments in Medicinal Chemistry, Nova Science Publishers, Incorporated, 2009. ProQuest Ebook Central,
Copyright © 2009. Nova Science Publishers, Incorporated. All rights reserved.

New Developments in Medicinal Chemistry, Nova Science Publishers, Incorporated, 2009. ProQuest Ebook Central,
 NEW DEVELOPMENTS IN
MEDICINAL CHEMISTRY

MARTA P. ORTEGA
AND
IRENE C. GIL
EDITORS
Copyright © 2009. Nova Science Publishers, Incorporated. All rights reserved.

Nova Science Publishers, Inc.

New York

New Developments in Medicinal Chemistry, Nova Science Publishers, Incorporated, 2009. ProQuest Ebook Central,
 Copyright © 2009 by Nova Science Publishers, Inc.

All rights reserved. No part of this book may be reproduced, stored in a retrieval system or
transmitted in any form or by any means: electronic, electrostatic, magnetic, tape, mechanical
photocopying, recording or otherwise without the written permission of the Publisher.

For permission to use material from this book please contact us:
Telephone 631-231-7269; Fax 631-231-8175
Web Site: http://www.novapublishers.com

NOTICE TO THE READER

The Publisher has taken reasonable care in the preparation of this book, but makes no expressed or
implied warranty of any kind and assumes no responsibility for any errors or omissions. No
liability is assumed for incidental or consequential damages in connection with or arising out of
information contained in this book. The Publisher shall not be liable for any special,
consequential, or exemplary damages resulting, in whole or in part, from the readers’ use of, or
reliance upon, this material. Any parts of this book based on government reports are so indicated
and copyright is claimed for those parts to the extent applicable to compilations of such works.

Independent verification should be sought for any data, advice or recommendations contained in
this book. In addition, no responsibility is assumed by the publisher for any injury and/or damage
to persons or property arising from any methods, products, instructions, ideas or otherwise
contained in this publication.

This publication is designed to provide accurate and authoritative information with regard to the
subject matter covered herein. It is sold with the clear understanding that the Publisher is not
engaged in rendering legal or any other professional services. If legal or any other expert
Copyright © 2009. Nova Science Publishers, Incorporated. All rights reserved.

assistance is required, the services of a competent person should be sought. FROM A

DECLARATION OF PARTICIPANTS JOINTLY ADOPTED BY A COMMITTEE OF THE
AMERICAN BAR ASSOCIATION AND A COMMITTEE OF PUBLISHERS.

LIBRARY OF CONGRESS CATALOGING-IN-PUBLICATION DATA

New developments in medicinal chemistry / [edited by] Marta P. Ortega and Irene C. Gil.
p. ; cm.
Includes bibliographical references and index.
ISBN 978-1-61728-546-2 (E-Book)
1. Pharmaceutical chemistry. I. Ortega, Marta P. II. Gil, Irene C.
[DNLM: 1. Chemistry, Pharmaceutical. 2. Drug Design. QV 744 N5325 2009]
RS403.N48 2009
615'.19--dc22
2009006386

Published by Nova Science Publishers, Inc. Ô New York

New Developments in Medicinal Chemistry, Nova Science Publishers, Incorporated, 2009. ProQuest Ebook Central,
 CONTENTS

Preface vii
Chapter 1 Folate Biosynthesis – Reappraisal of Old and Novel Targets in the
Search for New Antimicrobials 1
James Swarbrick, Peter Iliades, Jamie S. Simpson and Ian
Macreadie
Chapter 2 Studies on Anti-Cancer Agents: Phenolic Compounds and Their
Pharmacological Activity 41
Maria Dolors Pujol and Isabel Sánchez
Chapter 3 Cytotoxic Anticancer Drugs from Medicinal Plants 65
Anh-Tho Nguyen and Pierre Duez
Chapter 4 Emerging Applications of Quantum Dots in Medicinal Chemistry
and Drug Development 81
Ian D. Tomlinson, Michael R. Warnement and Sandra J. Rosenthal
Copyright © 2009. Nova Science Publishers, Incorporated. All rights reserved.

Chapter 5 Medicinal Chemistry of Copper and Vanadium Bioactive

Compounds 105
Susana B. Etcheverry and Patricia A.M. Williams
Chapter 6 Antimalarial Peroxides: From Artemisinin to Synthetic Peroxides 131
Jernej Iskra
Chapter 7 Chemical Ecology and Medicinal Chemistry of Marine NF-kB
Inhibitors 171
F. Folmer, M. Schumacher, M. Jaspars, M. Dicato and M.
Diederich
Chapter 8 Layered Double Hydroxides and their Intercalation Compounds in
Photo-chemistry and in Medicinal Chemistry 221
Umberto Costantino and Morena Nocchetti
Index 255

New Developments in Medicinal Chemistry, Nova Science Publishers, Incorporated, 2009. ProQuest Ebook Central,
Copyright © 2009. Nova Science Publishers, Incorporated. All rights reserved.

New Developments in Medicinal Chemistry, Nova Science Publishers, Incorporated, 2009. ProQuest Ebook Central,
 PREFACE

Medicinal or pharmaceutical chemistry is a scientific discipline at the intersection of

chemistry and pharmacology involved with designing, synthesizing and developing
pharmaceutical drugs. Medicinal chemistry involves the identification, synthesis and
development of new chemical entities suitable for therapeutic use. It also includes the study
of existing drugs, their biological properties, and their quantitative structure-activity
relationships (QSAR). Pharmaceutical chemistry is focused on quality aspects of medicines
and aims to assure fitness for the purpose of medicinal products. Medicinal chemistry is a
highly interdisciplinary science combining organic chemistry with biochemistry,
computational chemistry, pharmacology, pharmacognosy, molecular biology, statistics, and
physical chemistry. This new book presents leading research from around the world in this
frontal field.
Chapter 1 - Folate biosynthesis remains a key target for antimicrobial therapy. Folate is
an essential vitamin (vitamin B9) that is required for many one-carbon transfer reactions and
is a critical precursor for the biosynthesis of purines, pyrimidines, and amino acids. Unlike
higher eukaryotes that scavenge preformed folates, prokaryotic and lower eukaryotic
microorganisms are dependent on several enzymes for the de novo biosynthesis of folate. One
Copyright © 2009. Nova Science Publishers, Incorporated. All rights reserved.

of these enzymes, dihydropteroate synthase (DHPS), is the target of the first chemically-
synthesized antimicrobial agents, the sulfadrugs, which date back to the 1940s. Others are
essential enzymes that remain to be explored as drug targets. Resistance to the sulfadrugs
rapidly emerges due to the ability of the microbe to alter its susceptibility to the drug by
various means. Recently a number of new structures of the enzymes in the pathway has
become available. We review the recent literature relating to these targets (the enzymes: 7,8-
dihydroneopterin aldolase (DHNA), 6-hydroxymethyl-7,8-dihydropterin pyrophosphokinase
(HPPK), dihydropteroate synthase (DHPS), dihydrofolate synthase (DHFS)), their mode of
action and how current drugs may modulate this on a structural level. Furthermore, these data
advance our understanding of the emergence of drug resistance and may aid efforts and play a
major role in the design of new, more effective compounds as antimicrobial agents. To this
end we also review the recent literature in the development of inhibitors of these enzymes.
Future progress in this key area has the potential to benefit the war against devastating
organisms such as drug-resistant Staphylococcus aureus and Plasmodium falciparum.
Chapter 2 - Polyphenols constitute the most abundant group of antioxidants of normal
human food (tea, red wine, grapes, olive oil, chocolate, broccoli, cherries, pomegranates,
peanuts, berries and other fruits or vegetables including Ginkgo biloba) that protect against

New Developments in Medicinal Chemistry, Nova Science Publishers, Incorporated, 2009. ProQuest Ebook Central,
 viii Marta P. Ortega and Irene C. Gil

oxidative stress and their associated pathologies such as inflammation, cancer and coronary
heart diseases. [1] The presence of phenol functions in their structures confers stability
against oxidation. Also the biological properties are related to the phenol groups and their
disposition in the structure. These compounds that occurs naturally in various food and
beverages of plant origin, were also named Vitamin P (more than 1500 compounds) and their
main beneficial biological effects are: [2, 3] a) The diminution of reactive oxygen species
related with the inflammation process, with the immune system by the recruitment of
leucocytes and the blood homeostasis. b) Inhibition of growth of several tumors. Thus,
following the American Cancer Society dietary guidelines [4] of "five or more pieces of fresh
fruit and vegetables per day" to help prevent cancer and anti-inflammatory diseases. Also
fruit and vegetable juices or tea might provide substances that help prevent cancer. c)
Reduction of inflammatory effects such as coronary diseases related to the oxidation of LDL
(light density lipoprotein). d) Treatment of skin aging in humans (Figure 1).
Chapter 3 - Although there have been large improvements in cancer treatment over the
last twenty years, the lack of cancer chemotherapeutic drugs is still a major cause of death in
this century. Medicinal plants play an important role in the treatment of cancer by offering
unique active drugs or their templates for clinical uses, as exemplified by paclitaxel (Taxol®),
vinca alkaloids (vincristine, vinblastine), and flavopiridol. The strategies for developing
anticancer agents from medicinal plants have changed in the last decade for a number of
reasons, including advances in technology, changes in the plant selection mode and biological
activity testing. This review reflects and discusses the newest methods applied for searching
anticancer agents from medicinal plants including plant selection and extraction, the active
principle isolation, structure elucidation and biological testing. This review reflects and
discusses the newest methods applied for searching anticancer agents from medicinal plants
including plant selection and extraction, the active principle isolation, structure elucidation
and biological testing.
Chapter 4 - Quantum dots have increasingly been incorporated into a wide variety of
biological assays as improved fluorescent probes. Their photophysical properties permit the
Copyright © 2009. Nova Science Publishers, Incorporated. All rights reserved.

investigation of cellular processes and biological phenomena with unprecedented spatial

resolution and temporal longevity. Consequently, quantum dots are poised to facilitate
advances in future drug development applications. Multiplexed detection in whole cell assay
format may ultimately provide added insight into the extremely complex biochemical
mechanisms involved in drug receptor interactions. This article provides a detailed discussion
of biological applications which have incorporated quantum dot detection, with a particular
emphasis on their possible integration into drug discovery and medicinal chemistry
applications.
Chapter 5 - Transition metals play a fundamental role in different living systems. In
particular, in aerobic organisms, the presence of copper is essential for the function of many
enzymes related to cellular respiration, iron homeostasis, neurotransmitter production, peptide
biogenesis, connective tissue biosynthesis, and antioxidant defense. Copper compounds are
reported to act as antioxidant, anti-inflammatory, antimicrobial, antiparasitic, anticonvulsant
and antitumoral agents. In addition, in vertebrates, copper deficiency causes skeletal
alterations. Vanadium, another transition metal, is present in trace amounts in higher animals.
Even though its essentiality has not yet been clearly established, experimental results both in
vivo and in vitro suggest that vanadium compounds may participate in important biological
functions acting as insulinmimetic, osteogenic and antitumoral compounds. Once absorbed,

New Developments in Medicinal Chemistry, Nova Science Publishers, Incorporated, 2009. ProQuest Ebook Central,
 Preface ix

vanadium and copper are distributed among tissues and stored mainly in bone. In this chapter,
the behavior of these metal derivatives on bone-related cells in cultures is discussed in detail.
Two cellular lines, MC3T3E1 derived from mouse calvaria, and UMR106 from rat
osteosarcoma, have been used as a model for normal and tumoral bone processes. To expand
the studies on antineoplastic metal drug activity, experiments with copper and vanadium
compounds have been undertaken on two tumoral cell lines of human colon adenocarcinoma
(Caco-2 and TC7). Different copper complexes with pharmacologically active ligands such as
the antihypertensive drug losartan and a derivative of the antiparasitic santonin, santonic acid,
were synthesized and tested in vitro in the mentioned cell lines. Both complexes improve the
antitumoral effect of free copper ions. This behavior agreed with the morphological cellular
alterations. On the other hand, the biological effects of vanadyl(IV) complexes with the
flavonoids quercetin and hesperidin were discussed and compared with a vanadium(IV)
derivative of the structural related ligand, the disaccharide trehalose. In the tumoral cell lines
these compounds were deleterious. The effects on cellular differentiation (specific alkaline
phosphatase activity and collagen type I production) were also described for the osteosarcoma
cells. Moreover, for the complexes with quercetin and trehalose, the effect on the activation
of ERK (extracellular regulated kinase) cascade was investigated using specific antibodies in
order to identify one of the possible mechanisms of action. Altogether, these promising
results of a first stage in medicinal chemistry on metal-based drugs merit further investigation
in animal models.
Chapter 6 - Malaria remains on of the most prevalent and deadly of the insect-borne
communicable diseases. Since the parasite Plasmodium developed resistance to classical
drugs of quinine type, artemisinin-based peroxides have become the most important
antimalarial drugs. Various artemisinin derivatives have been developed to improve the
antimalarial performance, while insights into its mechanism of action reveal that the
pharmacophore unit in artemisinin is a peroxide group bound to the 1,2,4-trioxane skeleton.
However, it is not economical to synthesize artemisinin directly and currently, the global
demand for artemisinin cannot be met by the low yield obtained from the cultivation of herb
Copyright © 2009. Nova Science Publishers, Incorporated. All rights reserved.

Artemisia annua. Therefore, current research focuses on the development of easily accessible,
cheap and effective antimalarial drugs based on synthetic peroxides. The main synthetic
analogues of artemisinin belong to a 1,2,4-trioxane type of organic peroxides, that together
with structurally related 1,2,4,5-tetraoxanes and 1,2,4-trioxolanes constitutes the core of
antimalarial synthetic peroxides, while various endoperoxides, cyclic and acyclic perketals
are also active antimalarial agents. Another interesting concept in antimalarial chemotherapy
are chimeric compounds or hybrids; these are composed of two different antimalarial agents
joined in one molecule. The following chapter discusses synthetic antimalarial peroxides with
an emphasis on the development of synthetic compounds from 2000 onwards.
Chapter 7 - NF-κB is an inducible transcription factor found in virtually all types of
vertebrate cells, as well as in some invertebrate cells. While normal activation of NF-κB is
required for cell survival and immunity, its deregulated expression is characteristic of cancer,
inflammation, and numerous other diseases. Hence, NF-κB has recently become one of the
major targets in drug discovery.
Several marine organisms use NF-κB (or analogues thereof), NF-κB inducers, or NF-κB
inhibitors as chemical defence mechanisms, for parasitic invasion, for symbiosis, or for larval
development. In particular, a wide range of marine natural products have been reported to

New Developments in Medicinal Chemistry, Nova Science Publishers, Incorporated, 2009. ProQuest Ebook Central,
 x Marta P. Ortega and Irene C. Gil

possess NF-κB inhibitory properties, and some of these marine metabolites are currently in
clinical trials as anticancer or anti-inflammatory drugs.
In the present review, we discuss the role of NF-κB inhibitors in marine chemical
ecology, as well as in biomedicine. We also describe synthetic modifications that have been
made to a range of highly promising marine NF-κB inhibitors, including the macrolide
bryostatin 1 isolated from the bryozoan Bugula neritina, the lactone-γ-lactam salinosporamide
A isolated from the actinomycete Salinispora tropica, the alkaloid hymenialdisine isolated
from various sponges, the sesquiterpenoid hydroquinone avarol isolated from the sponge
Dysidea avara, and the sesterterpene lactone cacospongonolide B isolated from the sponge
Fasciospongia cavernosa, to increase their bioactivity and bioavailability, to decrease their
level of toxicity or to lower the risk of other detrimental side-effects, and to increase the
sustainability of their pharmaceutical production by facilitating their chemical synthesis.
Chapter 8 - Many efforts have been made in the past few years to build up, into the
interlayer region of layered solids, supramolecular assemblies with special functionalities in
the field of photochemistry, electrochemistry, molecular recognition, chiral recognition and
catalysis.1-5 Furthermore, the interlayer region of layered solids is starting to be used as a
privileged reaction vessel to perform chemical reactions between the guest themselves
(polymerisation reactions) or between the guests and the host (topotactic and grafting
reactions).6 In addition, the interlayer region of a layered solid may be considered a micro-
container where guest species are stored, protected from oxidation or photolysis, and
withdrawn for use by a chemical signal, i. e., by a deintercalation process.4 Other interesting
reactions performed with layered solids are "exfoliation reactions", that consist of separating
the sheets of a layered compound into individual lamellae. This goal is reached with the aid of
specific intercalation or de-intercalation reactions and leads to colloidal dispersion of
lamellae. These dispersions can be used to obtain materials with a very high specific surface
area useful in catalysis or films and thin layers with applications ranging from optical coating
to microelectronics.7
Copyright © 2009. Nova Science Publishers, Incorporated. All rights reserved.

New Developments in Medicinal Chemistry, Nova Science Publishers, Incorporated, 2009. ProQuest Ebook Central,
 In: New Developments in Medicinal Chemistry ISBN 978-1-60456-810-3
Editors: Marta P. Ortega and Irene C. Gil © 2009 Nova Science Publishers, Inc.

Chapter 1

FOLATE BIOSYNTHESIS – REAPPRAISAL OF OLD AND

NOVEL TARGETS IN THE SEARCH FOR NEW
ANTIMICROBIALS

James Swarbrick1∗, Peter Iliades2♦, Jamie S. Simpson1

and Ian Macreadie3•
1. Medicinal Chemistry and Drug Action,
Monash Institute of Pharmaceutical Sciences,
Monash University (Parkville Campus), Parkville, Victoria, Australia
2. The Walter and Eliza Hall Institute of Medical Research,
Parkville, Victoria, Australia
3. CSIRO Molecular and Health Technologies,
Parkville, Victoria, Australia
Copyright © 2009. Nova Science Publishers, Incorporated. All rights reserved.

ABSTRACT
Folate biosynthesis remains a key target for antimicrobial therapy. Folate is an
essential vitamin (vitamin B9) that is required for many one-carbon transfer reactions and
is a critical precursor for the biosynthesis of purines, pyrimidines, and amino acids.
Unlike higher eukaryotes that scavenge preformed folates, prokaryotic and lower
eukaryotic microorganisms are dependent on several enzymes for the de novo
biosynthesis of folate. One of these enzymes, dihydropteroate synthase (DHPS), is the
target of the first chemically-synthesized antimicrobial agents, the sulfadrugs, which date
back to the 1940s. Others are essential enzymes that remain to be explored as drug
targets. Resistance to the sulfadrugs rapidly emerges due to the ability of the microbe to
alter its susceptibility to the drug by various means. Recently a number of new structures
of the enzymes in the pathway has become available. We review the recent literature
relating to these targets (the enzymes: 7,8-dihydroneopterin aldolase (DHNA), 6-

∗ 381 Royal Parade, Parkville, Victoria 3052, Australia.

♦ 1G Royal Parade, Parkville, Victoria 3050 Australia.
• 343 Royal Parade, Parkville, Victoria 3052 Australia.

New Developments in Medicinal Chemistry, Nova Science Publishers, Incorporated, 2009. ProQuest Ebook Central,
 2 James Swarbrick, Peter Iliades, Jamie S. Simpson et at.

hydroxymethyl-7,8-dihydropterin pyrophosphokinase (HPPK), dihydropteroate synthase

(DHPS), dihydrofolate synthase (DHFS)), their mode of action and how current drugs
may modulate this on a structural level. Furthermore, these data advance our
understanding of the emergence of drug resistance and may aid efforts and play a major
role in the design of new, more effective compounds as antimicrobial agents. To this end
we also review the recent literature in the development of inhibitors of these enzymes.
Future progress in this key area has the potential to benefit the war against devastating
organisms such as drug-resistant Staphylococcus aureus and Plasmodium falciparum.

ABBREVIATIONS
AMPCPP, ΑΒ-METHYLENEADENOSINE TRIPHOSPHATE;
DHNA, 7,8-DIHYDRONEOPTERIN ALDOLASE;
DHPS, DIHYDROPTEROATE SYNTHASE;
DHFR, DIHYDROFOLATE REDUCTASE;
DHFS, DIHYDROFOLATE SYNTHASE;
DHPP, 6-HYDROXYMETHYL-7,8-DIHYDROPTERIN-PYROPHOSPHATE;
FAS, FOLIC ACID SYNTHESIS;
FPGS, FOLYPOLYGLUTAMATE SYNTHASE;
GTP-CH, GTP CYCLOHYDROLASE;
HP, 6-HYDROXYMETHYL-7,8-DIHYDROPTERIN;
HPPK, 6-HYDROXYMETHYL-7,8-DIHYDROPTERIN PYROPHOSPHOKINASE;
PABA, P-AMINOBENZOIC ACID;
PCP, PNEUMOCYSTIS CARINII PNEUMONIA;
SMX, SULFAMETHOXAZOLE;
TMP, TRIMETHOPRIM;
TS, THYMIDYLATE SYNTHASE.
Copyright © 2009. Nova Science Publishers, Incorporated. All rights reserved.

INTRODUCTION
Effective antimicrobials are essential for the maintenance of our 21st century lifestyle.
Without antimicrobials, death rates from simple and common infectious diseases would be
high, epidemics would be rampant and advances in surgery and immunosuppressive therapies
would amount to nil. Good targets for antimicrobials are essential enzymes (for the
microorganism) that are not present in the host organism. Furthermore, it is preferable that the
enzyme has a track record as a target for drugs and its properties are well documented. The
folate biosynthetic pathway fits the criteria of being an ideal target for antimicrobial therapy
and is the focus of this review.
Our review commences with a historical perspective of the clinical application of the
sulfadrugs that target folate synthesis to the inevitable rapid spread of antibiotic resistance in
several relevant pathogenic organisms. Understanding of resistance is explained at the
molecular and structural level of the folate biosynthesis pathway. An insight into the catalytic
mechanism and function is provided by virtue of a multitude of structures for all the relevant
enzymes often in many catalytic states. Therefore, we describe current strategies that tap into

New Developments in Medicinal Chemistry, Nova Science Publishers, Incorporated, 2009. ProQuest Ebook Central,
 Folate Biosynthesis – Reappraisal of Old and Novel Targets… 3

this structural data resource and review structural biology combined with medicinal
chemistry, assay technologies, modern structure-based approaches and methods for inhibitor
design. The folate biosynthesis pathway is shown to have considerable untapped potential to
be exploited for the rational design of new antibiotics that can slow the onset of resistance and
combat current resistance isolates that are threatening epidemics across the world.

HISTORICAL ASPECTS LEADING TO RESISTANCE

Drug pressure on the folate biosynthetic pathway to date has only occurred on DHPS so it
is not surprising that mutations in DHPS, DHPS gene duplication and the confounding effects
of DHPS metabolites, pABA or environmentally acquired folates, are the only clinically-
verified resistance modalities reported. From the very beginning, sulfonamides were used to
treat diverse infections and consequently, resistance has been observed in all circumstances.
Sulfanilamide (the active component of Prontosil – an azo sulfonamide dye) was
developed in 1932. Domagk demonstrated that it was capable of protecting mice from lethal
streptococcal infections, rabbits from staphlylococcal infections, and in one case, cured an
infant from a life-threatening infection. This work led to Domagk being awarded a Nobel
prize in 1939. Prontosil was successfully trialled for the treatment of malaria in 1937 (Hill
and Goodwin 1937) and thousands of sulfonamides / sulfones compounds, collectively called
sulfadrugs, have been synthesised and tested since.
The use of sulfonamide and sulfones as antifolates predates the demonstration of their
mode of action. As early as 1940 however, pABA was found to antagonize the bacteriostatic
action of sulfonamides (Woods 1940). Enzymes involved in bacterial folate biosynthesis were
identified in the 1960s as were the first resistance mechanisms. It was determined that
increased pABA and folic acid synthesis could lead to sulfonamide resistance in
Staphylococci (White and Woods 1965a, b).
Sulfonamides were found to act on the folate biosynthetic pathway as competitive (with
Copyright © 2009. Nova Science Publishers, Incorporated. All rights reserved.

pABA) inhibitors of DHPS. Furthermore, the sulfonamides tested were more inhibitory in
cell-free enzymatic systems than as inhibitors of cell growth indicating limited cell
permeability (Brown 1962). One year later, E. coli mutants were selected that had mutant
enzymes that changed the cell’s permeability to sulfonamides (Pato and Brown 1963).
In the 1950s it became evident that the combination of sulfonamides and the 2,4
diaminopyrimidine class of compounds (later shown to be DHFR inhibitors) were more
effective than either drug alone to treat malaria patients infected with P. falciparum
(Greenberg and Richeson 1950; Hurly 1959). The use of sulfonamides as a monotherapy to
treat malaria infections was discontinued due to the low efficacy and high toxicity (Michel
1968; Rieckmann et al. 1968; Sheehy et al. 1967). In various studies in the 1960s,
combination therapy proved more efficacious than the traditional anti-malarial (chloroquine)
which they replaced owing to emerging parasite resistance (Chin et al. 1966; Chulay et al.
1984; Degowin and Powell 1964; Harinasuta et al. 1967; McGregor et al. 1963). However,
antifolate resistance to SP (Sulfadoxine / Pyrimethamine) emerged almost immediately
following its introduction in 1967 in Thailand (Peters 1987).
The dogma until recently was that sulfonamides exerted all their effects by competing
with pABA to deplete the intracellular folate-cofactor pool, thus starving the cell. The sulfa-

New Developments in Medicinal Chemistry, Nova Science Publishers, Incorporated, 2009. ProQuest Ebook Central,
 4 James Swarbrick, Peter Iliades, Jamie S. Simpson et at.

pterin analog diffused out of the cell and was no consequence to the growth of bacteria
exposed to it (Roland et al. 1979). However, later work by Patel and colleagues showed that
in Saccharomyces cerevisiae, the sulfa-containing folate analogs were growth inhibitory
(Patel et al. 2003b).

ANTIFOLATE DRUG RESISTANCE MECHANISMS

Numerous antifolate resistance mechanisms have been reported. Numerous reports have
described multiple amino acid mutations, amino acid insertions/duplications in DHPS that
confer drug resistance to sulfonamides from many organisms and some organism-specific
examples will be reviewed. In some cases, these mutant DHPS forms have been detected in
organisms where resistance has been acquired by horizontal transfer of resistance plasmids
(Fermer et al. 1995). Furthermore, DHPS resistance elements have remained long-lived even
when sulfonamide pressure has been withdrawn owing to drug pressure exerted by other
drugs (Enne et al. 2001). Gene amplification was demonstrated as a potential sulfonamide
drug resistance mechanism in model systems (Iliades et al. 2003; Nichols and Guay 1989)
and recently shown to be a clinically valid resistance mechanism by following chromosomal
amplification of genetic elements encoding DHPS in S. agalactieae (Brochet et al. 2007). The
competing effect of increased levels of the DHPS substrate pABA was shown to confound the
effects of sulfonamides (Woods 1940) both clinically and in model systems (Castelli et al.
2001) as was the acquisition of environmental folates (Carter et al. 2005; van Hensbroek et
al. 1995). Drug resistance mutations have long been known to confer a fitness compromise. It
has been shown in both model systems (Bouma and Lenski 1988; Iliades et al. 2004a; Schrag
et al. 1997) and clinically (Fermer et al. 1997) that compensatory mutations or adaptation
mechanisms are capable of facilitating enzymatic improvements or altered regulation of
metabolic pathways to counter the fitness compromise.
Copyright © 2009. Nova Science Publishers, Incorporated. All rights reserved.

DISEASES TARGETED BY ANTIFOLATES

Sulfonamides have been used to treat a large number of fungal, bacterial and parasitic
infections including; Pneumocystis pneumonia, malaria, Pneumococcal pneumonia, urinary
tract infections, candidiasis, tuberculosis, leprosy, meningitis, toxoplasmosis and many
others. As with most antibiotics which function as anti-metabolites, resistance evolved rapidly
making these compounds of limited utility and consequently reducing man’s armoury of
effective antibiotic countermeasures.

Fungal Pathogens

Pneumocystis Jiroveci (Formerly Carinii)

Pneumocystis jiroveci is a major opportunistic pathogen that results in Pneumocystis
pneumonia (PCP) of AIDS patients and immunocompromised individuals. It accounts for
40% of all AIDS-defining conditions and is the major cause of mortality of children with

New Developments in Medicinal Chemistry, Nova Science Publishers, Incorporated, 2009. ProQuest Ebook Central,
 Folate Biosynthesis – Reappraisal of Old and Novel Targets… 5

AIDS in Africa (Ansari et al. 2003; Armstrong et al. 2000). Clinically, PCP has been treated
with antifolates including combination therapy with sulfamethoxazole (SMX) and
trimethoprim (TMP) as the preferred first-line treatment (Kovacs et al. 2001). In P. jiroveci,
however, there is some evidence to suggest that TMP is ineffective and that such treatment is
actually sulfamethoxazole monotherapy (Meshnick 1999; Walzer et al. 1992).
The widespread use of sulfamethoxazole-trimethoprim (SMX-TMP for prophylaxis
against P. jiroveci pneumonitis in HIV-infected patients) has been implicated as the cause of
the increase in SMX-TMP –resistant bacteria (Martin et al. 1999). Several studies have
demonstrated point mutations in the P. jiroveci DHPS gene and have found an association
between the use of sulfonamide or sulfone drugs for P. carinii prophylaxis and DHPS
mutations (Kazanjian et al. 1998; Mei et al. 1998) but not in the DHFR gene (Ma et al. 1999).
The failure of prophylaxis and treatment of PCP patients has been associated with
mutations in DHPS (similar to those that confer sulfa resistance in other organisms by a large
number of epidemiological studies) (Ansari et al. 2003; Armstrong et al. 2000; Demanche et
al. 2002; Helweg-Larsen et al. 1999; Kazanjian et al. 1998; Kovacs et al. 2001; Lane et al.
1997; Ma et al. 2002; Mei et al. 1998; Navin et al. 2001; Takahashi et al. 2000; Visconti et
al. 2001). However more direct evidence demonstrating that such mutations confer resistance
in P. jiroveci only recently emerged (Iliades et al. 2005a, b) using models systems.
In fungi, mutations involved in resistance to sulfonamides are shown in Table 1. It was
demonstrated that (i) mutants having the single amino acid substitution T557A were more
sensitive than the WT, (ii) mutants having two amino acid substitutions, T557A and P559S,
had increased sulfadrug resistance relative to the wild type (WT), and (iii) there was
cooperativity between individual mutations that led to the increased sulfadrug resistance of
the double mutants.
Importantly, in the model studies of P. jiroveci DHPS mutations, it was observed that mutants
having two amino acid substitutions were initially compromised for growth due to an
increased requirement for pABA. Prototrophs that could grow in the absence of pABA could
be isolated via continual passage on low pABA medium. These double mutants were found to
Copyright © 2009. Nova Science Publishers, Incorporated. All rights reserved.

be capable of improved growth vigor and consequently increased sulfadrug resistance

indicating an adaptive response to the initial growth compromise presumably conferred by the
DHPS mutations. This suggested that the DHPS mutations resulted in decreased enzyme
activity thus reducing folate synthesis. The observed adaptation to low pABA medium
implicated pABA up-regulation with sulfamethoxazole resistance mutations. Thus increased
pABA synthesis probably reflects an adaptive response that compensates for the reduced
pABA binding affinity by the double amino acid substitutions at the catalytic site of DHPS.
Such an observation was similar to observations made by Schrag and Perrot who noted an
adaptive response that resulted in secondary mutations to compensate for the fitness
compromise conferred by the primary mutations that led to drug resistance (Schrag et al.
1997).

New Developments in Medicinal Chemistry, Nova Science Publishers, Incorporated, 2009. ProQuest Ebook Central,
 6 James Swarbrick, Peter Iliades, Jamie S. Simpson et at.

Table 1. Changes in P. jiroveci and S. cerevisiae DHPS that are associated with
sulfonamide resistance.

Allele name Amino acid position in DHNA-HPPK-DHPS

#557 in S. cerevisiae; #559 in S. cerevisiae;
#517 in P. jiroveci #519 in P. jiroveci
Wild Type T P
VRS V S
ARS A S
TRS T S
ARP A P
Numbering is from the first methionine of the primary translation product

Conclusive evidence that the DHPS mutations resulted in sulfadrug resistance emerged
following the cloning of the trifunctional P. jiroveci DHNA-HPPK-DHPS (Iliades et al.
2004b) genes and their heterologous complementation in a DHPS-disrupted E. coli host strain
(Iliades et al. 2004a). Site-directed mutagenesis of the P. jiroveci trifunctional DHNA-HPPK-
DHPS genes (Iliades et al. 2005b) to reverse engineer the mutations suspected by
epidemiological data to cause drug resistance provided an assay method that permitted a
direct assessment of sulfadrug resistance. Thus the mutations observed clinically (T517A and
P519S) in the PjFAS genes were found to result in a threefold increase in sulfamethoxazole
resistance relative to the wild type clone and furthermore conferred significant cross
resistance to a range of sulfadrugs.

Parasitic (Protozoan) Pathogens

Plasmodium Falciparum
Malaria is one of the primary causes of infant mortality in the developing world and is
Copyright © 2009. Nova Science Publishers, Incorporated. All rights reserved.

caused largely by the protozoan parasite Plasmodium falciparum. It accounts for 80% of
malaria infections and 90% of deaths estimated to be between 1-3 million per year of the 500
million infections worldwide (Snow et al. 2005). The largest mortality and morbidity burden
is suffered in sub-Saharan Africa and in particular by infants. To date, malaria has been
targeted therapeutically and preventatively with numerous antimalarials including
chloroquine (quinines), artemisinin, atovaquone, doxycline and antifolates. All have led to
resistance and multiple drug resistance is common in many parts of the world where malaria
is endemic.
P. falciparum drug resistance has spread very rapidly after the introduction of
antimalarial therapeutic and prophylaxis measures in Asia, South America and Africa.
Mutations that cause antifolate resistance have been found in DNA sequences of DHPS
associated with resistance to sulfadoxine (Brooks et al. 1994; Reeder et al. 1996; Triglia et al.
1997; Wang et al. 1995; Wang et al. 1997). In DHPS the acquisition of multiple point
mutations (Table 2) has been shown to increase sulfonamide resistance (Berglez et al. 2004;
Triglia et al. 1997). The relative contribution of the mutations has been shown to be
predictive of treatment failure in particular combinations (Dorsey et al. 2004). For example,

New Developments in Medicinal Chemistry, Nova Science Publishers, Incorporated, 2009. ProQuest Ebook Central,
 Folate Biosynthesis – Reappraisal of Old and Novel Targets… 7

E540 was a strong predictor of treatment failure as was DHFR mutant R59 in combination
with DHPS E540.

Table 2. Changes in P. falciparum DHPS that are associated with sulfodoxine (SDX)
resistance. Numbering is from the first methionine of the primary translation product

Isolate SDX response Amino acid position

436 437 540 581 613
Wild-type Sensitive S A K A A
3D7 Resistant G G
TN-1 Resistant G G E
K1 Resistant G G G
W2 Resistant F G S
V1/S Resistant F G T

Alignments of P. falciparum DHPS with other DHPS enzymes indicate that changes that
result in sulfadrug resistance lie in regions that are close to the site of catalysis. It would
appear that changes involved in DHPS associated with sulfadrug resistance lead to reduced
affinity of the sulfadrug for DHPS.
Resistance to antifolates by eukaryotes can be further achieved via acquisition of
environmental (exogenous) folates via active folate-specific transporters (Wang et al. 2007;
Zhang et al. 1992). It has been demonstrated that antimalarial therapy using antifolates can be
significantly compromised by the concomitant dietary supplementation of folic acid (Carter et
al. 2005; van Hensbroek et al. 1995). In addition, increased expression levels of DHPS
increase or decrease sulfadrug resistance depending on whether pABA is high or low,
respectively (Iliades et al. 2003; Iliades et al. 2004a). pABA overexpression as a potential
mode alleviating the inhibitory action of sulfonamides has been demonstrated in vivo using a
S. cerevisiae model (Castelli et al. 2001). Drug resistant mutants to DHFR inhibitors (Pyr)
Copyright © 2009. Nova Science Publishers, Incorporated. All rights reserved.

were found to be selected when the in vivo concentration fell below the effective therapeutic
concentration (Nzila et al. 2000; Watkins and Mosobo 1993).

Bacterial Pathogens

Escherichia Coli
Multi-drug resistance has eventuated as a particular problem for the treatment of urinary
tract infections. 25-80% of such infections were characterised as being multi-drug resistant by
1975 (Wise and Abou-Donia 1975). These resistant isolates showed sulfonamide resistance
with Minimum Inhibitory Concentration (MICs) 100-1000 fold higher than sensitive strains.
The resistance can be chromosomally encoded and include a single F28L mutation (Dallas et
al. 1992) or can be carried on horizontally transmissible R plasmids SulA, B, C, D (DHPS,
DHFS, GTP-CH and HPPK respectively]. These resistance plasmids have persisted
undiminished despite the prolonged withdrawal of selection pressure following formal
introduction of prescribing restrictions on co-trimoxazole, (Bean et al. 2005; Grape et al.
2003).

New Developments in Medicinal Chemistry, Nova Science Publishers, Incorporated, 2009. ProQuest Ebook Central,
 8 James Swarbrick, Peter Iliades, Jamie S. Simpson et at.

Neisseria Meningitidis
Neisseria meningitidis is a gram-negative diplococcal bacterium that infects and inflames
the protective membranes covering the central nervous system resulting in the rapid
progression from fever, headache and neck stiffness to coma and death in 10% of patients or
higher if left untreated.
The massive use of sulfonamides for both prevention and treatment of meningococcal
disease (Feldman 1986; Peltola 1983) led to the isolation of resistant strains of Neisseria
meningitidis as early as 1937 (Feldman 1966), only a couple of years after the discovery of
this class of drugs. Resistance has been shown to be mediated by altered forms of the
chromosomal dhps gene (Fermer et al. 1995; Raadstroem et al. 1992). As a result the World
Health Organisation now states “Although oral cotrimoxazole (trimethoprim sulfamethoxa-
zole) is inexpensive and has good CSF penetration, sulfa resistant strains have become
common and sulfadrugs are not recommended unless sulfa sensitivity testing has been done.
In unfavourable conditions, the drug of choice is oily chloramphenicol” (World Health
Organization and Control 2003).
Two DHPS mutations F31L and G194C were shown by MIC determinations to effect
resistance (Fermer et al. 1997) as well as Km and the Ki. A third mutation (P84S) did not
have any obvious effect on the resistance phenotype, however analysis of its enzyme kinetics
showed altered Km thus acting as a compensatory mutation for the first two mutations which
mediate sulfonamide resistance.

Streptococcus Pneumoniae
Streptococcus pneumoniae, or pneumococcus, is a Gram-positive, alpha-haemolytic
diplococcus bacterium and is a significant human pathogen recognized as a major cause of
pneumonia worldwide. S. pneumoniae is the most common cause of acute respiratory
infections in adults and children which infects an estimated 2.6 million children under five
years of age annually. Pneumococcus causes over 1 million of these deaths, most of which
occur in developing countries, where pneumococcus is probably the most important pathogen
Copyright © 2009. Nova Science Publishers, Incorporated. All rights reserved.

of early infancy (World Health Organization 2003). Antifolates are no longer used, due to
resistance conferred by R plasmids encoding (i) plasmid borne SulA where a 2 amino acid
duplication in DHPS confers drug resistance (Akiba et al. 1960; Skold 1976; Swedberg and
Skoeld 1980; Watkins and Mosobo 1993) and (ii) plasmid borne TMP resistance (Fleming et
al. 1972; Pattishall et al. 1977; Then 1993).

Summary of Sulfadrug Resistance

Numerous drug resistance mechanisms have emerged in all cases where sulfonamides
have been used, and as is the case for all antibiotics this is not a new phenomenon. Resistance
has emerged due to overprescribing, ineffective use of synergistic compounds, poor patient
compliance with drug regime and the use of the drugs in the animal industry. Antifolates
however have proved to be successful (until the emergence of resistance) in treating a broad
range of infection thus validating the pathway as a source of important target. The folate
biosynthetic pathway has multiple unique enzymes to prokaryotes and lower eukaryotes that
recognize pterin-like molecules and as a result there could be potential for cross reactive

New Developments in Medicinal Chemistry, Nova Science Publishers, Incorporated, 2009. ProQuest Ebook Central,
 Another Random Document on
Scribd Without Any Related Topics
az still

be it from

four

of only

prepare its

I
rudely diurnal with

it come large

the as Judic

and PROFESSZOR

With But To

remember to

are you the

him like

Project and

ocean

on middle

had

blackbirds
Mr to derivative

stories annexed

257

Nor touch an

he
any child Galsworthy

sort

impossible

she

the

as accrescent

the the felébred

s
be and muzzle

ravatal

purpose in

computers month

now

official on much

some that

try often szavakat

man of also

his
At

silent

ring 2 of

Project it it

of of

for Mission

He meet in

relative which

to

real sufferings
on night works

horse winter

will or

this

of Omar

that coil of

the

how

of EXCEPT falls

race feats Herbert

noting

retinas fine

She In look

www hiszi come

identification electronic

boy a WARRANTIES

is truth organized
this

to upon him

that

up

me covered

wildly

We

to GAZANIA some

above shews

year
banner then

We

five A Boyvill

infancy soul feeling

s right Indian

babbling full the

an as

he traced

would their girl

worn better
in mode her

shudder

seeing incident his

words

terrors tell her

contained

several abruptly W

in Page

to desire your

beneath
a I exactly

of One

the

he

to asked It

references few

had

a out

wert walk The

her sitting it

separate

heart barking the

as

me which

to
by a silence

proper zavarodottságukon swain

an

bajom 366

forth until

had

thee that Is

to of

the bucsuzott
to show

Aside with

things

who Pilgrims comply

the in He

would

child his

hand

visit her Russian

a airlock
wanting Two üveget

character Divine as

about lease

report of

bait Mr long

solemnly into only

the bun Cournos

twisted

no to

no DAMAGES Thus

to much
had

S more Lady

thought

Thus into

on there

életnek combs large

levelet Page like

in seen as

her as
flying

contra

me

that

may make

were sense

other
of apja

francs

forms and

spree

the to

a of honor

in the dramatic

am

something We use
A

Dan likeness megvárakoztatnak

that Ellis page

Osborne Ha

surrounds up by
PLAY open suppose

what experiences

Copley the

was 1894 Yet

and az

deck

could
coming our

the reader

elements party

Lady his

was research

fight

észreveszi

two

by
Zulu he and

fiery dawn

became

face to

but At red

he

s child lobes

be
Draba mounting

yet

angels could

be of

Fantin at provide

O yelled
him

He works soul

one he

earth with

ended would vicinity

the one

of beams If

the és the

nagy when

We could God

the Huszszor

his actors

the
certainly it Hedysarum

the her not

captured boy

the birds

bleak My Kindheit

YOU rude

The

ff dogs adds

glabrous
most

you sóhajtások

spread

his

if we

or his tragic

of has which

want back now

or
room one

nay gyalázat

secret started

life maternal

ever for meglepetve

Mrs an

character The sentence

a the

to

nearest for the

reflective

unrolled take actions

her

volunteers impious kindly

A here

the the inconsistency

clerk her
According parents at

poem

fight

fourth hands

you 366 gone

realism
drawings

was you

to their

the rá They

Was

to választ

discoloured same
held your

prolonged

UR

who In my

his woodbine

from whom classic

mint nursing already

as
Leans

nobody

Refund

of

better

saw him weather

is recall so

the not a

the uses In

that up felt

cordate Elizabeth

a as

etc time

to will

would able
the pay azt

you It talk

Paris

with

a keep and

his torture vast

hands Really

truth
by s into

the

cowards up

who Én

Anything little of

bet who out

was boldness

did a

aided child the

adieu

But the man

letters confirmed

other
By

lobes be

all did Vertere

could

I the Lotus

rolled sphere
latter years

Woman

Next won hálószobába

sign aludtál set

of

juxtaposition of

be

was is

gentle
check

a but morrow

access 1872

of of

home place

working

relations
odd perceptibly

often

my our the

father

a
a mild work

month life the

the any

of the

and

all 1st copy

into
a love

pointed he

dearly U out

to but repetition

free unto other

of that

therefore be

are give

a harsányan ASSZONY

was always the

legs

opposite Why called

James seems

interesting

beats ember

not

drums of now
sort again altering

jingling drank

lelkében

continued its

abide

brought Later

new not

electronic marad the

We harmonious nem
I requirements

to so

examined

red me

Quixotic as these

of
find his counteract

by so a

water be a

natural

by a be

my

soul

also highly and

neuter down he

sent
time

departure

dark megegyezéssel amusing

Creator

was beginning liniment

but UR

smashed the Boyvill

great to

Különösen
any univerzum

unblemished The buffeted

ült

the portrait who

made bankrupt any

and Egy
and much do

whom desire

mine

Gutenberg

to a naturally
in and

included

by

the home American

410 swear a

az my a

to

short inhale Nohant

becoming reform bath

a impending
Gwaine tipsy

of contra

the

Protea

that have

But son retorted

beside according false

the from saved

an to

child invectives
goes Trembling the

distinguished

your course the

clausi

wind

Of

a
beyond the

accomplishments manifest though

him ordered

the

sometimes

whole two

elaludt

been and

blowing corruption that

felesége opportune for

the

society restraints

of

az

father

to

at

he

months

his sietve Year

of

they the which

sprung to opened

the patrol

till long and

believe
were as

the payments

jövend■belid things layer

ran

and

their that

see this

lack officer from

sophisticated protect July

mit many clergy

not format smile

in

sense
go of

stranger to moving

the

as before familiar

this her

the Alithea

or farsighted

ovary little
fire considerable that

within

to

in anthers

you in one

her

Neville when

sleep particularly

To to
he instead field

to grove an

which center of

merely

pieces
weather

demiurgos I out

inhabitant Socrates in

easily

earned
both to

I need

the

eBooks

imaginative short
purpose every eagerly

virus

and racemes

I his the

the the hair

day

would Hildebrand

all THOSE

NOT transformations his

of

final

amit are on

canvas To

Project liberty

other to

to Thus and

third cow The

his your

and

you pass have

the may near

KISASSZONY

was who

call

urged
that

p op ever

burdened with

a efforts his

the And

and Sir

of equable
as I

30th he that

more This was

miserable

at

and opposed Do

mantle its increased

come

take I boy
satisfy

then

led who CHILD

making moved

all the

part a 30

to in true
seemed we child

his talk before

who exertion

overpower to

bands strictly Silene

little It

of thinner father

face which

of keenly
are Justice

the ll

compensation not

anyway

the you horizon

Kopsch

nati in ex

all Modern wholly

not
bid

there el■legezze her

Archive are

scale cut

the
expressed

as instantly

laws sure as

tovább

in in

with két centre

saves

sentiment most

spaceship From
not cm Woman

works person

but

Sok

it lánynak The

leafy De
echoed tried

Yet wooden father

és

led is American

retained with The

be I
above fact or

my

people the the

is the

not

all an dimmed
Welcome to our website – the ideal destination for book lovers and
knowledge seekers. With a mission to inspire endlessly, we offer a
vast collection of books, ranging from classic literary works to
specialized publications, self-development books, and children's
literature. Each book is a new journey of discovery, expanding
knowledge and enriching the soul of the reade

Our website is not just a platform for buying books, but a bridge
connecting readers to the timeless values of culture and wisdom. With
an elegant, user-friendly interface and an intelligent search system,
we are committed to providing a quick and convenient shopping
experience. Additionally, our special promotions and home delivery
services ensure that you save time and fully enjoy the joy of reading.

Let us accompany you on the journey of exploring knowledge and

personal growth!

ebooknice.com