Pharmaceutical Biology 1388-0209/01/390S-008$16.

00
2001, Vol. 39, Supplement, pp. 8–17 © Swets & Zeitlinger

Natural Product Drug Discovery in the Next Millennium

Gordon M. Cragg and David J. Newman

Natural Products Branch, Developmental Therapeutics Program, Division of Cancer Treatment and Diagnosis, National
Cancer Institute, Fairview Center, Frederick, MD, USA
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Abstract Medicinals for the millennia

Nature has been a source of medicinal agents for thousands Recorded history
of years, and an impressive number of modern drugs have
Throughout the ages, humans have relied on nature for their
been isolated from natural sources, many based on their use
basic needs: the production of foodstuffs, shelters, clothing,
in traditional medicine. In the past century, however, an
means of transportation, fertilizers, flavors and fragrances,
increasing role has been played by microorganisms in
and not least, medicines. Plants have formed the basis of
the production of antibiotics and other drugs for the
sophisticated traditional medicine systems that have been in
treatment of some serious diseases. Advances in the descrip-
existence for thousands of years (Anonymous, 1998). The
tion of the human genome, as well as the genomes of
first records, written on clay tablets in cuneiform, are from
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pathogenic microbes and parasites, is permitting the deter-

Mesopotamia and date from about 2600 BC. Among the
mination of the structures of many proteins associated
substances which the Mesopotamian people used, were oils
with disease processes. With the development of new
of the Cedrus (cedar) and Cupressus sempevirens (cypress)
molecular targets based on these proteins, there is an increas-
species, Glycyrrhiza glabra (licorice), Commiphora species
ing demand for novel molecular diversity for screening.
(myrrh), and Papaver somniferum (poppy juice), all of which
Natural products will play a crucial role in meeting this
are still in use today for the treatment of ailments ranging
demand through the continued investigation of world’s bio-
from coughs and colds to parasitic infections and inflamma-
diversity, much of which remains unexplored. With less than
tion. Egyptian medicine dates from about 2900 BC, but the
1% of the microbial world currently known, advances in pro-
best known Egyptian pharmaceutical record is the “Ebers
cedures for microbial cultivation and the extraction of
Papyrus” dating from 1500 BC. This documents some 700
nucleic acids from environmental samples from soil and
drugs (mostly plants), and includes formulas, such as gargles,
marine habitats, will provide access to a vast untapped reser-
snuffs, poultices, infusions, pills, and ointments, made with
voir of genetic and metabolic diversity. The same holds true
beer, milk, wine, and honey as vehicles. The Chinese Materia
for nucleic acids isolated from symbiotic and endophytic
Medica has been extensively documented over the centuries,
microbes associated with terrestrial and marine macro-
with the first record dating from about 1100 BC (Wu Shi Er
organisms. By use of combinatorial chemical and biosyn-
Bing Fang, containing 52 prescriptions), followed by works
thetic technology, novel natural product leads will be
such as the Shennong Herbal (~100 BC; 365 drugs), and the
optimized on the basis of their biological activities to yield
Tang Herbal (659 AD; 850 drugs). Likewise, documentation
effective chemotherapeutic and other bioactive agents. The
of the Indian Ayurvedic system dates from about 1000 BC
investigation of these resources requires multi-disciplinary,
(Susruta and Charaka); this system formed the basis for the
national, and international collaboration in the discovery and
primary text of Tibetan Medicine, Gyu-zhi (Four Tantras),
development process.
translated from Sanskrit during the eighth century AD
(Fallarino, 1994).
Keywords: Collaboration, combinatorial biosynthesis/chem- In the ancient Western world, the Greeks contributed sub-
istry, drug discovery, molecular targets, natural products, stantially to the rational development of the use of herbal
synthesis. drugs. The philosopher and natural scientist, Theophrastus

Address correspondence to: Gordon M. Cragg, Natural Products Branch, Developmental Therapeutics Program, Division of Cancer
Treatment and Diagnosis, National Cancer Institute, Fairview Center, Room 206, P. O. Box B, Frederick, MD 21702-1201, USA.
Fax: (301)-846-6178, E-mail: cragg@dtpax2.ncifcrf.gov
 Natural product drug discovery
(~300 BC), in his “History of Plants”, dealt with the me-
dicinal qualities of herbs and noted the ability to change their
characteristics through cultivation. Dioscorides, a Greek
physician (100 AD), accurately recorded the collection,
storage, and use of medicinal herbs during his travels with
Roman armies throughout the then “known world”; he is con-
sidered by many to be the most important representative of
the science of herbal drugs in “ancient times”. Galen
(130–200 AD), who practiced and taught pharmacy and med-
icine in Rome, and published no less than 30 books on these
subjects, is well known for his complex prescriptions and for-
mulas used in compounding drugs, sometimes containing
dozens of ingredients (“galenicals”). During the Dark and
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Middle Ages (fifth to twelfth centuries), the monasteries in
countries such as England, Ireland, France, and Germany,
preserved the remnants of this Western knowledge. However,
it was the Arabs who were responsible for the preservation
of much of the Greco-Roman expertise, and for expanding it
to include the use of their own resources along with Chinese
and Indian herbs unknown to the Greco-Roman world. The
Arabs were the first to establish privately owned drug stores
in the eighth century, and the Persian pharmacist, physician,
philosopher, and poet, Avicenna, contributed much to the sci-
ences of pharmacy and medicine through works such as
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Canon Medicinae, regarded as “the final codification of all
Greco-Roman medicine”.
Traditional medicine and drug discovery
As mentioned above, plants have formed the basis for tradi-
tional medicine systems that have been used for thousands
of years in countries such as China (Chang & But, 1986) and
India (Kapoor, 1990). The use of plants in the traditional
medicine systems of many other cultures has been exten-
sively documented (Schultes & Raffauf, 1990; Arvigo &
Balick, 1993; Gupta, 1995; Ayensu, 1981; Iwu, 1993; Jain,
1991). These plant-based systems continue to play an essen-
tial role in health care, and it has been estimated by the World
Health Organization that approximately 80% of the world’s
inhabitants rely mainly on traditional medicines for their
primary health care (Farnsworth et al., 1985). Plant products
also play an important role in the health care systems of the
remaining 20% of the population, mainly residents of devel-
oped countries. Analysis of data on prescriptions dispensed
from community pharmacies in the United States from 1959
to 1980 indicates that about 25% contained plant extracts or
active principles derived from higher plants. Also, at least
119 chemical substances, derived from 90 plant species, can
be considered as important drugs currently in use in one or
more countries (Farnsworth et al., 1985). Of these 119 drugs,
74% were discovered as a result of chemical studies directed
at the isolation of the active substances from plants used in
traditional medicine.
The isolation of the antimalarial drug, quinine, from the
bark of Cinchona species (e.g., C. officinalis) was reported
in 1820 by the French pharmacists, Caventou and Pelletier.
9
The bark had long been used by indigenous groups in the
Amazon region for the treatment of fevers, and was first
introduced into Europe in the early 1600s for the treatment
of malaria. Quinine formed the basis for the synthesis of the
commonly used antimalarial drugs, chloroquine and meflo-
quine. Another plant long used in the treatment of fevers in
traditional Chinese medicine, Artemisia annua (Quinhaosu),
has yielded the agents, artemisinin and its derivatives –
artemether and artether, effective against strains of malaria
resistant to quinine and quinine-derivatives (Buss & Waigh,
1995). The analgesic used in ancient Mesopotamia (vide
infra), morphine, was isolated from the opium poppy
(Papaver somniferum) in 1816 by the German pharmacist,
Serturner. This isolation laid the basis for alkaloid chemistry,
and the development of a range of highly effective analgesic
agents (Buss & Waigh, 1995). In 1785, the English physi-
cian, Withering, published his observations on the use of the
foxglove, Digitalis purpurea, for the treatment of heart dis-
orders; this eventually led to the isolation of the cardiotonic
agent, digoxin.
There are many other significant drugs developed from
traditional medicinal plants, a few of which are mentioned
here: The antihypertensive agent isolated from Rauwolfia
serpentina, reserpine, has been used in Ayurvedic medicine
for the treatment of snakebite and other ailments (Kapoor,
1990). Ephedrine, first isolated in 1887 from Ephedra sinica
(Ma Huang), was used in traditional Chinese medicine,
and became the basis for the synthesis of the anti-asthma
agents (beta agonists) salbutamol and salmetrol. The muscle
relaxant, tubocurarine, isolated from Chondrodendron
and Curarea species, was used by indigenous groups in the
Amazon as the basis for the arrow poison, curare (Buss
& Waigh, 1995). The importance of traditional medicinal
plants, and the potential new drug leads that they represent,
are readily apparent.
The Golden Age of antibiotics
The serendipitous discovery of penicillin from the filamen-
tous fungus Penicillium notatum by Fleming in 1929, and the
observation of the broad therapeutic use of this agent in the
1940s, ushered in a new era in medicine and the “Golden
Age” of antibiotics. This discovery also had the effect of pro-
moting the intensive investigation of nature as a source of
novel bioactive agents. Microorganisms are a prolific source
of structurally diverse bioactive metabolites and have yielded
some of the most important products of the pharmaceutical
industry. These include: antibacterial agents, such as the
penicillins (from Penicillium species), cephalosporins (from
Cephalosporium acremonium), aminoglycosides, tetracy-
clines and polyketides (all from Streptomyces species);
immunosuppressive agents, such as the cyclosporins and
rapamycin (from Streptomyces species); cholesterol-
lowering agents, such as mevastatin (compactin) and lovas-
tatin (from Penicillium species); as well as anthelmintics and
antiparasitic drugs, such as the ivermectins (from Strepto-
 10 G.M. Cragg and D.J. Newman
myces species) (Buss & Waigh, 1995). A recent publication
reports the isolation of a potential antidiabetic agent from a
Pseudomassaria fungal species found in the rainforests of
the Congo (Zhang et al., 1999).
Marine sources
While marine organisms do not have a history of use in tra-
ditional medicine, the ancient Phoenicians employed a
chemical secretion from marine molluscs to produce purple
dyes for woolen cloth, and seaweeds have long been used to
fertilize soil. The world’s oceans, covering more than 70% of
the earth’s surface, represent an enormous resource for the
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discovery of potential chemotherapeutic agents. All but two
of the 28 major animal phyla are represented in aquatic envi-
ronments, with eight being exclusively aquatic and mainly
marine (McConnell et al., 1994). Prior to the development of
reliable scuba diving techniques some forty years ago, the
collection of marine organisms was limited to those obtain-
able by skin diving. Subsequently, depths from approxi-
mately ten feet to 120 feet became routinely attainable, and
the marine environment has been increasingly explored as a
source of novel bioactive agents. Deep water collections can
be made by dredging or trawling, but these methods suffer
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from disadvantages, such as environmental damage and non-
selective sampling. These disadvantages can be partially
overcome by use of manned submersibles or remotely oper-
ated vehicles (ROVs); however, the high cost of these forms
of collecting precludes their extensive use in routine collec-
tion operations.
The pseudopterosins, isolated from the Carribbean gor-
gonian, Pseudopterogorgia elisabethae, possess significant
analgesic and anti-inflammatory activity; defined fractions
obtained from extracts of the gorgonian are used topically in
skin lotions. Another marine product, manoalide, showing
potent anti-inflammatory activity, is isolated from the sponge
Luffarriella variabilis (McConnell et al., 1994) and has led
to a family of similar compounds via synthesis, some of
which have reached clinical trial status. The extremely potent
venoms (conatoxins) of predatory cone snails (Conus
species) have yielded complex mixtures of small peptides
(six to 40 amino acids long) which have provided models for
the synthesis of novel painkillers (e.g., Ziconotide) (Olivera,
1997).
Other sources
Teprotide, isolated from the venom of the pit viper, Bothrops
jaracaca, led to the design and synthesis of the ACE
inhibitors, captropril and enalapril (Buss & Waigh, 1995),
used in the treatment of cardiovascular disease. Meanwhile,
epibatidine, isolated from the skin of the poisonous frog,
Epipedobates tricolor, has led to the development of a novel
class of painkillers (Daly, 1998).
This interest in nature as a source of potential chemother-
apeutic agents continues. An analysis of the number and
sources of anticancer and anti-infective agents, reported
mainly in the Annual Reports of Medicinal Chemistry from
1984 to 1995 covering the years 1983 to 1994, indicates that
over 60% of the approved drugs developed in these disease
areas are of natural origin (Cragg et al., 1997a).
Anticancer agents from natural sources
Of the 92 anticancer drugs commercially available prior to
1983 in the United States, and approved worldwide between
1983 and 1994, approximately 62% can be related to natural
origins (Cragg et al., 1997a).
Plant sources
Plants have a long history of use in the treatment of cancer
(Hartwell, 1982), though many of the claims for the efficacy
of such treatment should be viewed with some skepticism
because cancer, as a specific disease entity, is likely to be
poorly defined in terms of folklore and traditional medicine
(Cragg et al., 1994). Of the plant-derived anticancer drugs in
clinical use, the best known are the so-called vinca alkaloids,
vinblastine and vincristine, isolated from the Madagascar
periwinkle, Catharanthus roseus. C. roseus was used by
various cultures for the treatment of diabetes, and vinblastine
and vincristine were first discovered during an investigation
of the plant as a source of potential oral hypoglycemic agents.
Therefore, their discovery may be indirectly attributed to the
observation of an unrelated medicinal use of the source plant.
The two clinically-active agents, etoposide and teniposide,
which are semisynthetic derivatives of the natural product
epipodophyllotoxin, may be considered as more closely
linked to a plant originally used for the treatment of cancer.
Epipodophyllotoxin is an isomer of podophyllotoxin which
was isolated as the active antitumor agent from the roots of
various species of the genus Podophyllum. These plants
possess a long history of medicinal use by early American
and Asian cultures, including the treatment of skin cancers
and warts (Cragg et al., 1994).
More recent additions to the armamentarium of naturally-
derived chemotherapeutic agents are the taxanes and camp-
tothecins. Paclitaxel was initially isolated from the bark of
Taxus brevifolia, collected in Washington State as part of a
random collection program by the U.S. Department of Agri-
culture for the National Cancer Institute (NCI) (Cragg et al.,
1993). The use of various parts of T. brevifolia and other
Taxus species (e.g., canadensis, baccata) by several Native
American tribes for the treatment of some non-cancerous
conditions has been reported (Cragg et al., 1994). The leaves
of T. baccata are used in the traditional Asiatic Indian
(Ayurvedic) medicine system (Kapoor, 1990), with one
reported use in the treatment of cancer (Hartwell, 1982).
Paclitaxel, along with several key precursors (the baccatins),
occurs in the leaves of various Taxus species; the ready semi-
synthetic conversion of the relatively abundant baccatins to
 Natural product drug discovery
paclitaxel, as well as active paclitaxel analogs (such as doc-
etaxel) (Cortes & Pazdur, 1995), has provided a major renew-
able natural source of this important class of drugs. Likewise,
the clinically-active agents, topotecan (hycamptamine),
irinotecan (CPT-11), and 9-amino- and 9-nitro-camptothecin,
are semi-synthetically derived from camptothecin isolated
from the Chinese ornamental tree, Camptotheca acuminata
(Potmeisel & Pinedo, 1995). Camptothecin (as its sodium
salt) was advanced to clinical trials by the NCI in the 1970s,
but was dropped because of severe bladder toxicity.
Microbial sources
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Antitumor antibiotics are amongst the most important of the
cancer chemotherapeutic agents, which include members of
the anthracycline, bleomycin, actinomycin, mitomycin, and
aureolic acid families (Foye, 1995). Clinically useful agents
from these families are the daunomycin-related agents,
daunomycin itself, doxorubicin, idarubicin, and epirubicin;
the glycopeptidic bleomycins A2 and B2 (blenoxane); the pep-
tolides exemplified by dactinomycin; the mitosanes such
as mitomycin C; and the glycosylated anthracenone,
mithramycin. All were isolated from various Streptomyces
species. Other clinically active agents isolated from Strepto-
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myces include streptozocin and deoxycoformycin.
Marine sources
The first notable discovery of biologically-active compounds
from marine sources was the serendipitous isolation of the
C-nucleosides, spongouridine and spongothymidine, from
the Caribbean sponge, Cryptotheca crypta, in the early
1950s. These compounds were found to possess antiviral
activity; synthetic analog studies eventually led to the devel-
opment of cytosine arabinoside (Ara-C) as a clinically useful
anticancer agent, together with Ara-A as an anti-viral,
approximately 15 years later (McConnell et al., 1994). The
systematic investigation of marine environments as sources
of novel biologically active agents only began in earnest in
the mid-1970s. During the decade from 1977–1987, about
2500 new metabolites were reported from a variety of marine
organisms. These studies have clearly demonstrated that the
marine environment is a rich source of bioactive compounds,
many of which belong to totally novel chemical classes not
found in terrestrial sources (Carte, 1996).
As yet, no compound isolated from a marine source has
advanced to commercial use as a chemotherapeutic agent,
though several are in various phases of clinical development
as potential anticancer agents. The most prominent of these
is bryostatin 1, isolated from the bryozoan, Bugula neritina
(McConnell et al., 1994). This agent exerts a range of bio-
logical effects, thought to occur through modulation of
protein kinase C, and has shown some promising activity
against melanoma in Phase I studies (Philip et al., 1993). The
sea hare, Dolabella auricularia from the Indian Ocean, is the
source of more than 15 cytotoxic cyclic and linear peptides,
11
the dolastatins. The most active of these is the linear tetrapep-
tide, dolastatin 10, which has been chemically synthesized
and is currently in Phase I clinical trials (Carte, 1996).
Sponges are traditionally a rich source of bioactive com-
pounds in a variety of pharmacological screens (Carte,
1996); in the cancer area, halichondrin B, a macrocyclic
polyether initially isolated from the sponge Halichondria
okadai in 1985, is currently in pre-clinical development by
the NCI. Halichondrin B and related compounds have been
isolated from several sponge genera, and the present source
– a Lissodendoryx species, is being successfully grown by
in-sea aquaculture in New Zealand territorial waters (Cragg
et al., 1997b). The mechanisms of action of discodermolide
(Haar et al., 1996), isolated from the Caribbean sponge Dis-
codermia sp., and eleutherobin (Long et al., 1998), isolated
from a Western Australian soft coral Eleutherobia sp., are
similar to that of paclitaxel, with the former now in pre-
clinical development with Novartis.
Development of molecular targets and high
throughput screens
With the rapid progress in the sequencing of the human
genome comprising an estimated 30,000 genes, a vast
amount of information is becoming available which is
leading to a better understanding of human diseases in terms
of biology, diagnosis, prevention, and treatment. Knowledge
of the genes associated with the onset of diseases enables the
identification of the proteins expressed by these genes. These
proteins may serve as molecular targets for the development
of high throughput assays for the testing of thousands of
materials, including natural products, some of which may act
as inhibitors in the progression of the relevant diseases. It is
estimated that about one to ten thousand protein targets may
be identified through the human genome project. In addition,
the sequencing of the genomes of pathogens and parasites
will permit the identification of the genes essential for the
survival of the pathogens, and their encoded proteins may
serve as molecular targets for drug discovery.
Over the past 20 years, there has been an explosion in the
understanding of how cancer cells work. Through the Cancer
Genome Anatomy Project (CGAP: www.ncbi.nlm.nih.gov/
cgap/), it is the goal of the NCI to identify as many of the
human genes associated with cancer as possible. Through
gene sequence analysis, numerous mutational sites in cancer
cells have been, and are being, identified, some of which are
unique to specific types of cancer. This knowledge permits
the prediction of the structure of the encoded proteins asso-
ciated with the malignant process, and the discovery of pos-
sible molecular targets affecting important aspects of cancer
cell function. Anti-cancer drugs which have emerged from
molecular target approaches, are being evaluated in the
clinic, and include inhibitors of angiogenesis, farnesyl tran-
ferase, signal transduction, metalloprotease, protein kinase
(PK) antagonists, and modulators of gene expression (anti-
sense oligonucleotides).
 12 G.M. Cragg and D.J. Newman
A specific example is the development of a promising new
oral agent, STI-571, for the treatment of chronic myeloge-
nous leukemia (CML). CML is associated with an abnor-
mality resulting from the exchange of segments of
chromosomes 9 and 22 (called the Philadelphia chromo-
some). This exchange produces a member of the tyrosine
kinase family called bcr-abl. Screening of compound
libraries at the drug company Ciba-Geigy (now Novartis) for
specific inhibition of bcr-abl led to the identification of STI-
571. Patients treated with doses of 140 mg or greater have
had significant hematologic responses, defined as greater
than 50% decreases in white blood cell counts sustained for
periods exceeding two weeks. Additional trials of STI-571
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are planned for patients with advanced-stage CML, as well
as with acute lymphocytic leukemia associated with the
Philadelphia chromosome (Anonymous, 1999).
The ultimate goals envisaged are the creation of an inte-
grated, a cohesive drug discovery program, an early clinical
trials system that is founded on mechanistic-based
approaches, and to make emerging knowledge of cancer
biology the basis for drug discovery, development, and
testing.
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Generation of molecular diversity
Exploration of new environments
The potential of the marine environment as a source of novel
drugs has already been discussed. The NCI contract collec-
tion program has been expanded to the waters off East and
Southern Africa, and expansion to under-explored regions,
such as the Red Sea, is being considered. These collections
are performed in close collaboration with organizations
based in the countries controlling the relevant waters.
Exciting untapped resources are the deep-sea vents occur-
ring along ocean ridges, such as the East Pacific Rise and the
Galapagos Rift. Exploration of these regions is being per-
formed by several organizations, including the Center for
Deep-Sea Ecology and Biotechnology of the Institute of
Marine and Coastal Sciences at Rutgers University. The
center is using deep-sea submersibles such as Alvin to cata-
logue the rich biological resources of deep-sea macro and
microorganisms (Lutz & Kennish, 1993; Lutz et al., 1994).
Samples are being evaluated by the NCI in collaboration with
chemists at Research Triangle Institute.
Despite the more intensive investigation of terrestrial
flora, it is estimated that only 5–15% of the approximately
250,000 species of higher plants have been systematically
investigated, chemically and pharmacologically (Balandrin et
al., 1993). The potential of large areas of tropical rainforests
remains virtually untapped and may be studied through col-
laborative programs with source country organizations, such
as those established by the NCI.
Another vast untapped resource is that of the insect world,
and organizations such as the Instituto Nacional de Biodi-
versidad (INBio) in Costa Rica are investigating the poten-
tial of this resource in collaboration with some pharmaceu-
tical companies and the NCI.
The continuing threat to biodiversity through the destruc-
tion of terrestrial and marine ecosystems lends an urgency to
the need to expand the exploration of these resources as a
source of novel bioactive agents.
The unexplored potential of microbial diversity
Until recently, microbiologists were greatly limited in their
study of natural microbial ecosystems due to an inability to
cultivate most naturally occurring microorganisms. In a
report recently released by the American Academy of Micro-
biology entitled, “The Microbial World: Foundation of the
Biosphere”, it is estimated that “less than 1% of bacterial
species and less than 5% of fungal species are currently
known.” Other recent evidence also indicates that millions of
microbial species remain undiscovered (Young, 1997).
The recent development of procedures for cultivating and
identifying microorganisms will aid microbiologists in their
assessment of the earth’s full range of microbial diversity. In
addition, procedures based on the extraction of nucleic acids
from environmental samples will permit the identification
of microorganisms through the isolation and sequencing of
ribosomal RNA or rDNA (genes encoding for rRNA).
Samples from soils are currently being investigated, and the
methods may be applied to other habitats, such as the
microflora, insects, and marine animals (Handelsman et al.,
1998). Valuable products and information are certain to result
from the cloning and understanding of the novel genes which
will be discovered through these processes.
Extreme habitats harbor a host of extremophilic microbes
(extremophiles), such as acidophiles (acidic sulfurous hot
springs), alkalophiles (alkaline lakes), halophiles (salt lakes),
baro- and thermophiles (deep-sea vents) (Persidis, 1998), and
psychrophiles (arctic and antarctic waters, alpine lakes)
(Psenner & Sattler, 1998). While investigations thus far have
focused on the isolation of thermophilic and hyperther-
mophilic enzymes (Adams & Kelly, 1998), there are reports
of useful enzymes being isolated from other extreme habi-
tats (www.diversa.com). These extreme environments will
also undoubtedly yield novel bioactive chemotypes.
As Dr. Rita Colwell, Director of the United States
National Science Foundation, commenting on the impor-
tance of exploration and conservation of microbial diversity
has stated: “Hiding within the as-yet undiscovered microor-
ganisms are cures for diseases, means to clean polluted envi-
ronments, new food sources, and better ways to manufacture
products used daily in modern society” (Colwell, 1997).
Combinatorial biosynthesis
Advances in the understanding of bacterial aromatic polyke-
tide biosynthesis have lead to the identification of multi-
functional polyketide synthase enzymes (PKSs) responsible
for the construction of polyketide backbones of defined
 Natural product drug discovery
chain lengths, the degree and regiospecificity of ketoreduc-
tion, and the regiospecificity of cyclizations and aromatiza-
tions, together with the genes encoding for the enzymes
(Hutchinson, 1999). Since polyketides constitute a large
number of structurally-diverse natural products exhibiting a
broad range of biological activities (e.g., tetracyclines, dox-
orubicin, and avermectin), the potential for generating novel
molecules with enhanced known bioactivities, or even novel
bioactivities, appears to be high (Gokhale et al., 1999).
The NCI is promoting this area of research through the
award of grants to consortia, composed of multidisciplinary
groups devoted to the application of combinatorial biosyn-
thetic and/or combinatorial chemical techniques, for the
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generation of molecular diversity for testing with high
throughput screens related to cancer.
Total synthesis of natural products
The total synthesis of complex natural products has long
posed challenges to the top synthetic chemistry groups
worldwide, has led to the discovery of many novel reactions,
and has guided developments in chiral catalytic reactions
(Service, 1999). More recently, the efforts of some groups
have been focused on the synthesis and modification of drugs
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that are difficult to isolate in sufficient quantities for devel-
opment. In the process of total synthesis, it is often possible
to determine the essential features of the molecule necessary
for activity (the pharmacophore). In some instances, this has
led to the synthesis of simpler analogs having similar or
better activity. Notable examples in the anticancer drug area
are the synthesis of synthetic analogs of the marine organ-
ism metabolites, bryostatin 1 (Wender et al., 1998) and
ecteinascidin 743 (Martinez et al., 1999).
The synthesis of the epothilones by several groups has
permitted the preparation of a large number of designed
analogs and detailed structure-activity studies which are
reviewed in an excellent recent article (Nicolaou et al.,
1998a). These studies have identified desirable modifications
that might eventually lead to more suitable candidates for
drug development, but thus far, none of the analogs has sur-
passed epothilone B in its potency against tumor cells.
The similarity in the mechanisms of action of paclitaxel,
the epothilones, discodermolide and eleutherobin has led
to proposals that these structurally dissimilar substances
possess common pharmacophores which could lead to the
design and synthesis of analogs having substantially differ-
ent structures and superior activities (Borman, 1999).
Combinatorial chemistry and natural products
In the study of the structure-activity relationships of the
epothilones, solid-phase synthesis of combinatorial libraries
has been used to probe regions of the molecule important to
the retention and improvement of activity (Nicolaou et al.,
1998a). The combinatorial approach, using an active natural
product as the central scaffold, can also be applied to the gen-
13
eration of large numbers of analogs for structure-activity
studies, the so-called parallel synthetic approach (Nicolaou
et al., 1998b).
The split-and-pool solid-phase synthetic approach has
also been used to assemble a library of over 2 million natural
product-like compounds from 18 chiral tetracyclic scaffolds,
30 terminal alkynes, 62 primary amines, and 62 carboxylic
acids, using a six-step reaction sequence (Schreiber et al.,
1998). This library will be used to probe complex biological
processes, including protein-protein interactions, for which
no ligands have as yet been identified. This approach of
probing complex biological processes by altering the func-
tion of proteins through binding with small molecules has
been called chemical genetics (Schreiber, 1998).
Collaboration in drug discovery and
development: the NCI role
Much of the NCI drug discovery and development effort has
been, and continues to be, carried out through collaborations
with academic institutions, research organizations, and the
pharmaceutical industry worldwide. Many of the naturally
derived anticancer agents were developed through such
efforts. The DTP/NCI thus complements the efforts of the
pharmaceutical industry and other research organizations
through taking positive leads, which the pharmaceutical
industry might consider too uncertain to sponsor, and con-
ducting the high-risk research necessary to determine their
potential utility as anticancer drugs. In promoting drug dis-
covery and development, the DTP/NCI has formulated
various mechanisms for establishing collaborations with
research groups worldwide.
Source country collaboration
Drug discovery: memorandum of understanding
As discussed, the collections of plants and marine organisms
have been carried out in over 25 countries through contracts
with qualified botanical and marine biological organizations
working in close collaboration with qualified source country
organizations. The recognition of the value of natural
resources (plant, marine, and microbial) being investigated
by the NCI, and the significant contributions being made by
source country scientists in aiding the performance of the
NCI collection programs, have led the NCI to formulate its
Letter of Collection (LOC). This letter specifies policies
aimed at facilitating collaboration with, and compensation
of, countries participating in the drug discovery program
(Mays et al., 1997).
With the increased awareness of genetically-rich source
countries to the value of their natural resources, and the con-
firmation of source country sovereign rights over these
resources by the U.N. Convention of Biological Diversity,
organizations involved in drug discovery and development
are increasingly adopting policies of equitable collaboration
 14 G.M. Cragg and D.J. Newman
and compensation in interacting with these countries (Baker
et al., 1995). Particularly in the area of plant-related studies,
source country scientists and governments are committed to
performing more of the operations in-country, as opposed to
simply exporting raw materials. The NCI has recognized this
fact for several years, and has negotiated Memoranda of
Understanding (MOU) with a number of source country
organizations suitably qualified to perform in-country pro-
cessing. In considering the continuation of its plant-derived
drug discovery program, the NCI has de-emphasized its
contract collection projects in favor of expanding closer
collaboration with qualified source country scientists and
organizations. In establishing these collaborations, the NCI
Pharmaceutical Biology Downloaded from informahealthcare.com by University of Laval on 06/26/14
undertakes to abide by the same policies of collaboration and
compensation as specified in the LOC. A number of other
organizations and companies have implemented similar
policies (Baker et al., 1995). Through this mechanism col-
laborations have been established with organizations in
Bangladesh, Brazil, China, Costa Rica, Fiji, Iceland, Korea,
Mexico, New Zealand, Nicaragua, Pakistan, Panama, South
Africa, and Zimbabwe.
Drug development
For personal use only.
The calanolides
In 1988, an organic extract of the leaves and twigs of the tree
Calophyllum lanigerum, collected in Sarawak, Malaysia in
1987, through the NCI contract with the University of Illinois
at Chicago (UIC) and in collaboration with the Sarawak
Forestry Department, showed significant anti-HIV activity.
Bioassay-guided fractionation of the extract yielded (+)-
calanolide A as the main in vitro active agent (Kashman
et al., 1992). Attempted recollections in 1991 failed to locate
the original tree, and collections of other specimens of the
same species gave only trace amounts of calanolide A. In
1992, a detailed survey of C. lanigerum and related species
was undertaken by UIC and botanists of the Sarawak Forestry
Department. As part of the survey, latex samples of Calo-
phyllum teysmanii were collected and yielded extracts
showing significant anti-HIV activity. The active constituent
was found to be (-)-calanolide B which was isolated in yields
of 20 to 30%. While (-)-calanolide B is slightly less active
than (+)-calanolide A, it has the advantage of being readily
available from the latex which is tapped in a sustainable
manner by making small slash wounds in the bark of mature
trees without causing any harm to the trees. A decision was
made by the NCI to proceed with the pre-clinical develop-
ment of both the calanolides, and, in June of 1994, an agree-
ment based on the NCI Letter of Collection was signed
between the Sarawak State Government and the NCI. Under
the agreement, a scientist from the University of Malaysia
at Sarawak was invited to visit the NCI laboratories in
Frederick to participate in the further study of the compounds.
The NCI obtained patents on both calanolides, and in
1995, an exclusive license for their development was
awarded to Medichem Research, Inc., a small pharmaceuti-
cal company based near Chicago. Medichem Research
had developed a synthesis of (+)-calanolide A (Flavin et al.,
1996) under a Small Business Innovation Research
(SBIR) grant from the NCI. The licensing agreement
specified that Medichem Research negotiate an agreement
with the Sarawak State Government. Meanwhile, by late
1995, the Sarawak State Forestry Department, UIC, and
the NCI had collaborated in the collection of over 50 kg
of latex from C. teysmanii, and kilogram quantities of
(-)-calanolide B have been isolated for further development
towards clinical trials. Medichem Research, in collabora-
tion with the NCI through the signing of a Cooperative
Research and Development Agreement (CRADA) by which
NCI is contributing research knowledge and expertise, has
advanced (+)-calanolide A through pre-clinical development.
The collaborative group was granted an INDA for clinical
studies by the U. S. Food and Drug Administration (FDA).
The Sarawak State Government and Medichem Research
formed a joint venture company in late 1996, Sarawak
Medichem Pharmaceuticals Incorporated (SMP), and SMP
has sponsored Phase I clinical studies with healthy volun-
teers. It has been shown that doses exceeding the expected
levels required for efficacy against the virus are well toler-
ated. Phase II trials using patients infected with HIV-1 are
in progress.
The development of the calanolides is an excellent
example of collaboration, in the development of promising
drug candidates, between a source country (Sarawak,
Malaysia), a company (Medichem Research, Inc.), and the
NCI. It illustrates the effectiveness and strong commitment
of the NCI to policies promoting the rights of source coun-
tries to fair and equitable collaboration and compensation in
the drug discovery and development process. The develop-
ment of the calanolides has been reviewed as a “Benefit-
Sharing Case Study” for the Executive Secretary of the
Convention on Biological Diversity by staff of the Royal
Botanic Gardens, Kew (Ten Kate & Wells, 1998).
Screening agreement
In the case of organizations, such as pharmaceutical and
chemical companies or academic research groups, that wish
to have pure compounds tested in the NCI drug screening
program, the DTP/NCI has formulated a screening agree-
ment which includes terms stipulating confidentiality, patent
rights, routine and non-proprietary screening and testing
versus non-routine, and levels of collaboration in the drug
development process. Individual scientists and research orga-
nizations wishing to submit pure compounds for testing gen-
erally consider entering into this agreement with the NCI
DCTD. Should a compound show promising anticancer
activity in the routine screening operations, the NCI will
propose the establishment of a more formal collaboration,
such as a Cooperative Research and Development Agreement
(CRADA) or a Clinical Trial Agreement (CTA).
 Natural product drug discovery
DTP WWW Homepage
The NCI DTP offers access to a considerable body of
data and background information through its WWW
Homepage: http://dtp.nci.nih.gov. Publicly available data
include results from the human tumor cell line screen,
an AIDS antiviral drug screen, the expression of molecular
targets in cell lines, and 2D and 3D structural information.
Background information is available on the drug screen
and the behavior of standard agents, NCI investigational
drugs, analysis of screening data by COMPARE, the
AIDS antiviral drug screen, and the 3D database. Data and
information are only available on so-called “open com-
pounds” which are not subject to the terms of confidential
Pharmaceutical Biology Downloaded from informahealthcare.com by University of Laval on 06/26/14
submission.
In providing screening data on extracts, they are identi-
fied by code numbers only; details of the origin of the
extracts, such as source organism taxonomy and location of
collection, may only be obtained by individuals or organiza-
tions prepared to sign agreements binding them to terms of
confidentiality and requirements regarding collaboration
with, and compensation of, source countries. Such require-
ments are in line with the NCI commitments to the source
countries through its LOC.
For personal use only.
Conclusions
As illustrated in the foregoing discussion, nature is an
abundant source of novel chemotypes and pharmacophores.
However, it has been estimated that only 5 to 15% of
the approximately 250,000 species of higher plants have
been systematically investigated for the presence of bioactive
compounds (Balandrin et al., 1993), while the potential
of the marine environment has barely been tapped
(McConnell et al., 1994). The Actinomycetales have
been extensively investigated and have become a major
source of novel microbial metabolites (Horan, 1994).
However, less than 1% of bacterial and less than 5% of fungal
species are currently known, and the potential of novel
microbial sources, particularly those found in extreme envi-
ronments, seems unbounded. To these natural sources can be
added the potential to investigate the rational design of novel
structure types within certain classes of microbial metabo-
lites through genetic engineering, as has been elegantly
demonstrated with bacterial polyketides. The proven natural
product drug discovery track record, coupled with the con-
tinuing threat to biodiversity through the destruction of ter-
restrial and marine ecosystems, provides a compelling
argument in favor of expanded exploration of nature as a
source of novel leads for the development of drugs and other
valuable bioactive agents. It is apparent that nature can
provide the novel chemical scaffolds for elaboration by com-
binatorial approaches (chemical and biochemical), thus
leading to agents that have been optimized on the basis of
their biological activities.
15
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