Natural products isolated from higher plants arid microorganisms

have been providing novel, clinically active drugs. The key to the success of discovering naturally
occurring therapeutic agents rests on bioassay-guided fractionation and purification procedures.

Screening of both synthetic organic compounds and extracts of natural products has had an impressive
history of identifying active agents. For example, there are about 50 commercially available anticancer
drugs (excluding endocrines) which have been approved to date by the USFDA, and significantly, the
drugs based on natural products represent almost 1/3 of these total approved agents. A most recent
addition is taxol (approved in 1992, and the semi-synthetic in 1995), a natural product derived from the
Pacific yew tree Taxus brevifolia, which is used for the treatment of ovarian and breast cancer.

Malaysia has about 12,000 species of flowering plants of which about 1,300 are said to be medicinal
(Burkill, 1935), and only about a hundred have been investigated fully for their potential. The huge
diversity of the Malaysian flora means that we can expect well diversed chemical structures from their
secondary metabolises, and chemical diversity is one of the plus factors that makes natural products
excellent candidates for any screening programme.

The current trend of forest destruction for the sake of development means that scientists and the
science involved will have to progress even faster in order to get at the potential natural products from
the remaining forests. Universiti Kebangsaan Malaysia (UKM) and other local universities and research
institutes are embarking on their own programmes which normally involves a concerted
multidisciplinary approach in the discovery of bioactive agents from plant-derived natural products.
Certain novel strategies have been carried out based on the expertise and funds available at their
disposal and it is imperative that a screening programme be implemented and administered accordingly
with the ultimate aim of discovering new chemicals from the Malaysian flora for use in the
pharmaceutical and related industries. This in turn will help in the development and transfer of
technology in the said industries which can only be achieved through collaborative programmes with
other better equipped countries.

THE EARLY YEARS

The use of plants as medicinals by the local people have been well documented as early
as in 1935 by Burkill, recent books and papers on ethnobotany and ethnopharmacology are also
available but not as comprehensive. Medicinal plants were also available at that time in the markets or
side walks, mostly in crude form, sold by traditional medical practitioners. It was not until the 1950s that
any attempt was made to carry out scientific investigation on these medicinal plants although awareness
on the usefulness of such plants has been established for quite some time. Projects undertaken at that
time was simple and straightforward but effective which involved surveys of plant species available in a
certain designated area, their ethnobotanical use, and laboratory testing for certain classes of
compounds which were already known to be effective drugs. So it was not surprising then that most of
the phytochemical surveys carried out concentrated on the testing of alkaloids in these plants since
alkaloids were already (generally) proven to be effective as drugs.

The first report of a phytochemical survey of plants in Malaysia was carried out by Arthur in 1954, and
this was followed by several more similar reports right up to the 90s (Teo et al., 1990). This was, and still
is, a proven strategic approach whereby plants with alkaloids are chosen for further detailed
investigation. Inadvertently, a trend was set early on in Malaysia whereby most of the phytochemical
work concentrated on plants belonging to certain families or genus only because they contain alkaloids.

Some examples are the Annonaceae, Apocynaceae, Lauraceae, Menispermaceae, and Rubiaceae. There
was only one report ( Nakanishi et al., 1965) which made use of bioactivity as a means of selecting plants
for further investigation but this was not fully utilised until the present time (Said et al., 1995a; Latiff et
al., 1995). Even the ethnobotanical data, though well documented, was not fully utilised for this
purpose.

The main reason was because present-day bioassays were not available at that time, other bioactivity
tests, if available, were deemed expensive. Furthermore, Malaysia did not have enough manpower for a
multidisciplinary approach and scientists at that time worked more in isolation, a lack of funds also
made sure that such projects remain a low priority in a new nation.
Things began to improve when UNESCO got into the picture in the early 70s, funds were made available
for training of young scientists and regional cooperation became more coordinated where networking
was encouraged. Malaysia was actively involved in the Regional Network for the Chemistry of Natural
Products in Southeast Asia and through this network our scientists were exposed to current trends and
situations in the natural products area by interacting with other networks, notably from Australia
(NCBINP), Sweden (IFS), and Pakistan (ISESCO).

THE PRESENT STATE OF RESEARCH

The development of medicinal plants into therapeutic drugs takes several years and
millions of dollars are needed, hence making the process very capital-intensive the risks are also high
and the success rate not very good. Despite all this, natural products drug discovery programmes are
still in existence all over the world, mainly because of several reasons:

The high chemical diversity from natural products as compared to synthethics, the potential of these
natural products is largely unknown,

The large number of terestrial and marine species yet uninvestigated, and the back to nature
syndrome.

Modern technology and advancements made in this field in the last few years has made such
programmes attractive.

High throughput screens and sensitive instrumentation for structure elucidation has greatly reduced
the amount of time (and also the amount of sample) required for the first stage of the investigation.

Biotechnology is also expected to play a major role in the production of natural products through
biosynthesis and bioengineering which will further reduce the dependence on a large amount of plant
samples and hence reduce the pressure of reducing the number of biogenetic resources in our tropical
forests. But at the same time recent developments in the fields of combinatorial chemistry, high
throughput screening, computational capabilities and large data base management and manipulation
are beginning to shift interest away from natural products. Only time will tell whether this threath will
become a reality.
Malaysia, with her rich source of medicinal plants from the tropical forests, have taken steps to increase
the scientific knowledge of these plants by making available funds for research beginring in 1985 with
the IRPA (Intensification of Research in Priority Areas) programme. IRPA is now in its third term (1996-
2000) and one of the areas that have been identified as a priority is the commercialisation of
biotechnology, which also takes into account the development and production of biopharmaceu ticals
from plant genetic resources. Bioprospecting and phytochemistry would at best equip the country with
the necessary knowledge and a build up of a good data base that can be used to venture into the
capital-intensive programme of drug development with the right foreign partners.

There is a need for equitable sharing not only in the profits that may come out from such a venture but
also the necessary transfer of research knowledge and technology to the resource country. More than
five groups are now funded by IRPA to do phytochemical and bioscreening work but not all groups are
fully equipped or have the luxury of a multidisciplinary team. The overall direction is still not quite
obvious other than that determined in detail by that particular research group although coordination
between the different groups is good.

The emergence of phytomedicinals and nutraceuticals as an important alternative means of health care
world wide has also posed a dilemma: get involved with a drug discovery programme which is very
capital-intensive OR develop our own phytomedicinal industry which is already in existence?

The phytomedicine market in Europe is huge with more than US$6 billion in annual sales, and Germany
alone accounting for half of the market (Grunwald, 1995), while the US market is slightly less but with an
annual growth of about 12% (Brevoort, 1996) makes phytomedicinals an attractive business venture.
The Malaysian phytomedicinal market is worth about RMI billion and efforts have been made by the
Health Ministry to make their products more acceptable for export. This calls for more scientific input
through research by the same groups mentioned earlier. Malaysia is both involved in the development
of phytomedicines and the discovery of therapeutic drugs from biogenetic resources with the future in
mind.

SOME NOTABLE EXAMPLES

The six universities and two research institutes that are involved in natural products
research meet annually to discuss their fmdings and seek ways of improving the quality of their research
. The Malaysian Natural Products Society was formed in 1994 as a result of these meetings although the
annual meetings have been going on for the last twelve years. Other national committees have also
been formed to coordinate the many activities connected with medicinal plants, one of which is
responsible for the publication of a Malaysian Pharmacopea.

The University of Malaya was responsible for the initial work on Malaysian medicinal plants because
she was the only university then and the activities at that institution has continued to flourish until the
present time with more than three groups actively involved. The earlier publications concentrated more
on phytochemical work especially on alkaloids (for example Kiang et al., 1964) and the trend has
continued over the years until the present time (Chan et al, 1966 , 1986; Goh et al., 1985; Kam et al.,
1993, 1996). Very little work has been published on the bioactivity of all these alkaloids isolated
although some of them are known to have moderate cardiovascular effects (Chang et al., 1989), while
tumor promoting activities of some plant extracts were also investigated (lIham et al., 1996). In a way,
University of Malaya has gained a lot from their successful collaboration with the Institut de Chimie des
Substances Naturelles (CNRS-France) which started in 1982 and since renewed in 1993. This
collaboration has resulted in more than 50 publications, 6 PhD theses and 1 patent. At the onset,
however, the collaboration also concentrated only on the alkaloid-bearing plants (Clivio et al, 1990;
Jossang et al, 1991; Montacnac et al., 1995) but the emphasis has since shifted to the present day trend
of bioassay-directed isolation of natural products (Omobuwajo et al., 1 996; Alias et al., 1995).

The approach taken by the natural products group from Universiti Kebangsaan Malaysia has always
been a multi-disciplinary one (Latiff et al., 1984) although the methods used were a bit different from
their approach today. The phytochemists did their own thing by isolating nice crystalline compounds
which were then sent for bioactivity studies, and most of the time these compounds were not active.
The exercise was more or less to tag the compounds with some bioactivity studies. This approach was
later reversed , phytochemical work was only done on plant extracts that showed bioactivity and
compounds were isolated through bioassay-guided isolation techniques. Phytochemists were now
required to do simple benchtop bioassays (notably the brine shrimp lethality test) to help them isolate
the bioactive compounds which were then put through the other bioassays available to the group.

At the same time, the group did not abandon the traditional way of selecting plants through
phytochemical screening (Said et al., 1990, 1995a, 1995b) which made available plants that were not
collected or recorded before, and with the etnobotanical data available, the group was able to come in
contact with a large number of plant samples. The group has worked on the drug plant Mitragyna
speciosa for some time now and are still pursuing the bioactive compounds besides isolating some novel
ones (Houghton et al., 1991). Other notable examples are work on the anti-plasmodial plants Alstonia
anguslifolia (Said et al., 1992a; Wright et al., 1992) and Dehaasia incrassata (Said et al., 1991, 1992b)
which combined ethnobotanical data and bioassays to isolate the chemical compounds. The group also
has a special interest in the Annonaceae because of the large number of plants in that family that are
being used as traditional medicine, the group also has a taxanomist that specilizes in the Annonaceae.
Several species of Goniothalamus were investigated for their chemical components (Din et al., 1990;
Colgate et al., 1990; Said et al., 1995c) and also for their bioactivity (Rawarian et al., 1994).
Other than having a multidisciplinary team, the UKM group seems to be the only ones that are working
on the chemistry of lichens which has resulted in a series of papers on Malaysian lichens (Galloway, et
al., 1994). Novel compounds were isolated (Hamat et al., 1993; Samsudin et al., 1995), new species
discovered (Elix, et al., 1991; Din et al., 1995), but their bioactivies have been shown to be not really
significant (Din et al., 1992).

Other groups that have contributed significantly to the development of natural products research in
Malaysia, starting from the 1980's, are from Universiti Pertanian Malaysia (UPM), Universiti Teknologi
Malaysia (UTM), Universiti Sains Malaysia, (USM) and the Forest Research Institute of Malaysia (FRIM).
All are deeply involved in phytochemical work but are currently developing their own bioassay systems
with the exception of USM which has a School of Pharmacy to complement the chemical work. The
group in UTM has concentrated mainly on phytochemical work especially on the chemistry of the ginger
family (Sirat et al., 1996; Sirat, 1994).

The group in UPM is also leaning towards phytochemistry but have the simple bioassays at hand for
some bioactivity work (Hainzan et al., 1994; Rahmani et al., 1995). With the presence of a pharmacy
school, USM is able to do some anti-malarial work (Ang et al., 1995) and has also been involved in some
tissue culture work for the production of high yielding medicinal plants (Chan et al., 1995) The Forest
Research Institute of Malaysia is dedicated to the production of economically useful secondary forest
products and has formed a Medicinal Plants Division in 1995 for this purpose. Most of their work has
concentrated on the essential oils (Jantan et al., 1994, 1995) and their use in consumer products, but are
currently expanding into other classes of compounds and their bioactivities (Sahaari and Waterman,
1994; Connolly et al., 1996, Buchanan et al., 1996).

Special mention should also be given to Universiti Malaysia Sarawak (Unimas), although new, have been
actively involved with the natural products scene in Malaysia in the last couple of years. They have
developed an anti-termite screen (Sent et Al, 1994) and are also involved in the development of other
bioassays, besides being involved indirectly with the identification of several anti-HIV components from
the Calophyllum trees of Sarawak (Pengsuparp et al.., 1996). The future focus of natural products
research will be on Sabali and Sarawak due to the floral diversity of the two states.

PROSPECTS AND REALITIES

Biotechnology and Natural Products was the theme of the 37th Annual Meeting of the
American Society of Pharmacognosy held in July 1996 which indicates the importance of biotechnology
playing a major role in the search for biologically active natural products from plants and other sources.
Although natural products has the chemical diversity and has been proven to be useful templates, their
sustainable supply from the original biological source will always remain a problem. Obtaining the
biomes through bioengineering and the products themselves from biosynthesis would be a more
acceptable alternative. There have been many examples, but can biotechnology meet the challenges in
this particular problem when it has been successful in others? Combinatorial chemistry can produce a
variety of compounds and candidates for testing but at the present state will not be able to offer the
structural diversity that natural products can.

Malaysia has a small pool of scientists to work on a large flora, as such will not be able to focus on a
restricted area of research in natural products without compromising the freedom to do research. But,
even after forty plus years of research on the natural products of Malaysian flora, the nation has not
produced any compounds of pharmaceutical interest. Even in phytomedicinals, most of the raw
materials are imported from India, China and Indonesia, even though the market is getting bigger. There
are few venture capitalists that are willing to invest in the production of medicinal plants or for their oils
(Hunter, 1994), it is more lucrative (and sensible) to grow two of our biggest agricultural commodities,
rubber and oil palm.

Bioprospecting will remain an attractive proposition as long as we have the floral diversity, but there is
an urgent need to produce something more tangible out of this exercise in order to sustain the interest
and commitment of the government because only they are the ones that would be willing to invest on a
capital-intensive project. The prospects are there, the infrastructure and manpower is obtainable, the
business realities will have to be looked into more thoroughly.

CONCLUSION

Bioprospecting and the development of a natural products drug discovery programme is something that
the nation will have to address seriously. It is a long term capital-intensive programme that may or may
not bring any rewards, but with the technology and advancements made in the last few years, the
chances of success is good if the genetic biodiversity of the country is taken into account. The
programme will have to focus not only on the national needs but also on the global demands where the
market and finance is ever present.