Chapter1: Introduction

Chapter1: Introduction

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 Chapter1: Introduction

1.1: Introduction

The term of medicinal plants include a various types of plants used in

herbalism and some of these plants have a medicinal activities. These medicinal
plants consider as a rich resources of ingredients which can be used in drug
development and synthesis. Besides that these plants play a critical role in the
development of human cultures around the whole world. Moreover, some plants
consider as important source of nutrition (1). The definition of Medicinal Plant has
been formulated by WHO (World Health Organization) as follows- “A medicinal
plant is any plant which, in one or more of its organ, contains substance that can be
used for therapeutic purpose or which is a precursor for synthesis of useful drugs.”
Plants make many chemical compounds for biological functions including defense
against insects, fungi and herbivorous animals. Over 12000 active compounds are
known to science. As drug discovery from plants has traditionally been time-
consuming, faster and better methodologies for plant collection, bioassay screening,
compound isolation and compound development must be employed (2).

Biologically active compounds present in the medicinal plants have always been of
great interest to scientists. Phytochemical and pharmacological investigations of
plants is a huge potential research area where a lot of efforts are made and inputs are
yet to be discovered completely to make it a more affordable option for human
ailment. The phytochemical profiles of herbal remedies can provide hints for
designing, screening and developing novel multi target therapeutics. Nearly 80% of
the world’s population relies on traditional medicines for primary health care. Plants
can produce different kind of secondary metabolites also known as natural products as
they elicit effects on other organisms. For this reason the use of plant derived
medicine is increasing day by day.

As per data available over three-quarters of the world population relies mainly on
plants and plant extracts for their health care needs. More than 30% of the entire plant
species, at one time or other was used for medicinal purposes. It has been estimated,
that in developed countries such as United states, plant drugs constitutes as much as
25% of the total drugs, whiles in fast developing countries such as India and China the
contribution is as much as 80% [3].

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Many of the plants could be used as stimulants, poisons, hallucinogens or as medicine

because of the presence of unique or rich biological-active plant chemicals (i.e.
Chemical compounds that have a biological effect on another organism). Chemicals
that make a plant valuable as medicinal plant are Alkaloids (compounds has addictive
or pain killing or poisonous effect and sometimes help in important cures, Glycosides
(use as heart stimulant or drastic purgative or better sexual health), Tannins(used for
gastro- intestinal problems like diarrhea, dysentery, ulcer and for wounds and skin
diseases),Volatile/essential oils (enhance appetite and facilitate digestion or use as
antiseptic/insecticide and insect repellant properties), Fixed oils (present in seeds and
fruits could diminish gastric/acidity), Gum- resins and mucilage (possess analgesic
property that suppress inflammation and protect affected tissues against further injury
and cause mild purgative), and Vitamins and minerals (Fruits and vegetables are the
sources of vitamins and minerals and these are used popularly in herbals).In
Bangladesh 5,000 species of angiosperm are reported to occur (IUCN, 2003). The
numbers of medicinal plants included in the ‘materia medica’ of traditional medicine
in this subcontinent at present stands at about 2,000.More than 500 of such medicinal
plants have so far been enlisted as growing in Bangladesh. Dhaka, Rajshahi, Shylet
and Chittagong division is rich in medicinal plants.(4)

About40% of all modern medicines is either natural products or their semi-synthetic

derivatives, where erythromycin and rifampicin (bacterial infections), statins
(hyperlipidemia), quinines and artimesinin (malaria), paclitaxel, vinblastine and
vincristine (cancer), are a few well-known natural products-based medicines. For
bacterial infections, over 80% of all medicines in clinical use is either natural products
or their derivatives, while for anti- cancer agents over 60% of all drugs is either
natural products or derivatives thereof; examples of several potential lead molecules
are vincristine, vinblastine, taxol, camptothecin, podophyllotoxin, combretastatins, etc
[5)
which have been isolated from plants for successful use in cancer treatment . The
study of natural products has advantages over synthetic drug design in that it leads
optimally to materials having new structural features with novel biological activity.
Not only do plants continue to serve as important sources of new drugs, but
phytochemicals derived from them are also extremely useful as lead structures for
synthetic modification and optimization of bioactivity. Virtually every
pharmacological class of drugs includes a natural product prototype. Numerous
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methods have been utilized to acquire compounds for drug discovery, including
isolation from plants and other natural sources, synthetic chemistry, combinatorial
chemistry and molecular modeling[6)Different approaches to drug discovery from
plants can be enumerated as: random selection followed by chemical screening,
random selection followed by one or more biological assays, follow-up of biological
activity reports, follow-up of ethnomedical (traditional medicine) use of plants, use of
appropriate plant parts as such in powdered form or preparation of
enriched/standardized extracts (herbal product development), use of a plant product,
biologically potent but beset with other issues, as a lead for further chemistry, and
single new compounds as drugs.

Medicinal plants have been used extensively by a large proportion of the world for
their health care and remedy of diseases during the 2000 years and these data have
revealed a high degree of correlation between traditional medicinal plants uses and
laboratory analysis(7)

In the United States over the period 1999 to 2012, despite several hundred
applications for new drug status, only two botanical drug candidates had sufficient
evidence of medicinal value to be approved by the Food and Drug
Administration[8)Medicinal plants are widely used in non- industrialized societies,
mainly because they are readily available and cheaper than modern medicines [9)The
annual global export value of 50,000 to 70,000 plants with suspected medicinal
properties was estimated to be US$2.2 billion in 2012 and in2017, the potential global
market for botanical extracts and medicines was estimated at several hundred billion
dollars(10)

The Royal Botanic Gardens, Kew estimated in 2016 that 17,810 plant species have a
medicinal use, out of some30000 plants for which a use of any kind is documented [11).
By 2007, clinical trials had demonstrated potentially useful activity in nearly 16% of
herbal medicines; there was limited in vitro or in vivo evidence for roughly half the
medicines; there was only phytochemical evidence for around 20%; 0.5% were
allergenic or toxic; and some 12% had basically never been studied scientifically[12)

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Therefore it is necessary to perform extensive phytochemical and pharmacological

investigation for isolation and introduction of new medicinal entities for mankind to
combat the outbreak of new diseases and resistance of medicine. That is the
evaluation of different pharmacological activities of the methanolic extract of Grewia
nervosa swill satisfy the different ethno-pharmacological use of the plant and will
encourage further sresearch to investigate and isolate the lead compound for efficient
treatment of different diseases.

1.2 Rationale of the work

There is resurgence in the use of herbal medicines globally; hence, implying increased
efforts in evaluating these medicinal plants to establish their safety and efficacy [13)
Extensive phytochemical investigation and isolation of the active components in the
pure form thus become necessary to avoid untoward effects and to ensure safe use of
herbal medicines. Infectious diseases still represent an important cause of morbidity
[14)
and mortality among humans, especially in developing countries Even though
pharmacological industries have produced a number of new antibiotics in the last
three decades, resistance to these drugs by microorganisms has increased [15). Whereas
there are some advantages of using antimicrobial compounds of medicinal plants,
such as often fewer side effects, better patient tolerance, relatively less expensive,
[16)
acceptance due to long history of use and being renewable in nature .Medicinal
plants offer a number of bioactive compounds which may be good candidates for
further future drug development as- they can provide a good lead compound.
Outbreak of newer diseases and development of resistance are forcing the scientists to
widen this research. According to a recent research, scientists said that, we need at
least ten new medicines in each decade due to bacterial resistance.

The uncontrolled and abuse of antibiotics has resulted in increased antimicrobial

resistance and is seriously threatening drug control strategies against such common
diseases as tuberculosis, malaria, cholera, dysentery and pneumonia. In this
connection, medicinal plants offer a number of bioactive compounds which can be
good candidates for future drug development as-

 These can provide a good lead compound and drug discovery can be stemmed
in large proportion from the diverse structures.
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 Since secondary metabolites from natural sources have been elaborated within
living systems, they are often perceived as showing more “drug-likeness and
[17)
biologic friendliness than totally synthetic molecules,” making them good
candidates for further drug development.

A recent analysis by Fabricant and Farnsworth (2001) showed that the use of 80%
of 122 plant-derived drugs was related to their original ethno-pharmacological
purposes. In fact as this form of therapeutic practice passes on verbally from
generation to generation, proper record and documentations are not found in most
cases. So experiment should be carried out to identify bioactive compounds
responsible for pharmacological activity and new therapeutic value of these plants
other than the ethno-pharmacological use and work should be carried out for
proper documentation as well. Moreover, assessment of these phytomedicines is
also important including quality, stability, safety and efficacy which should also
be conducted in proper way after isolation of bioactive compound.

So, it is well visualized that phytochemical and pharmacological investigation of

plant is a huge potential research area where a lots of efforts, inputs are to be
given yet to discover it completely to make it a more affordable option for human
ailment.

This observation has driven the interest of the present research work towards
evaluation of phytochemical and pharmacological profile of an important
medicinal plant of Bangladesh “Grewia nervosa(Family-Malvaceae), locally
known as “Kathgua or Fattashi”.

1.3: Importance and current status of phytochemical research in Bangladesh

Bangladesh has been gifted with a rich plant diversity base because of its
heterogeneous ecologic condition such as fertile alluvial land, warm and humid
climate. There are about 6000 species of indigenous and naturalized plants growing in
the country. More than 1000 of these plants species in Bangladesh contain
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medicinally active chemical substances. (18) The practice of herbal medicine is

deeply rooted in the cultural heritage of Bangladesh and constitutes an integral part of
the culture of the people of this country. Different forms of traditional herbal
medicines have been used in this country as an essential means of treatment of
diseases and management of various health problems from time immemorial. The
practice of traditional medicine in this country has flourished
tremendously in the recent years along with that of modern medicine. As a result,
even at this age of highly advanced allopathic medicine, a large majority (75-80%) of
the population of this country, particularly in the rural and semi-urban areas, still
prefer to use traditional medicine in the treatment of most of their diseases even
though modern medical facilities may be available in the neighborhood.

As a developing country, herbal remedies contribute mostly in the health care system.
Homeopathy, Ayurveda and Unani systems are practiced in Bangladesh from long
time ago. Unani and Ayurvedic systems of medicine were officially recognized by the
Government of Bangladesh immediately after independence and at the same time a
Board of Unani and Ayurvedic systems of medicine was constituted. After the
introduction of a National Drug Policy in 1982, Unani and Ayurvedic drugs have been
brought under the control of the Drugs Administration Department of the Ministry of
Health and Family Welfare by legislation to control and regulate the commercial
manufacturing and marketing of quality Unani and Ayurvedic drugs. The Government
is also planning to incorporate traditional medicine in Primary Health Care (PHC)
activities. In order to achieve the goal of providing basic health needs to maximum of
the rural people in the shortest possible time with minimum expenditure, the
Government is planning to bring traditional medicine into the mainstream of the
organized public health services and health care delivery programs of the country
(19).
Medicinal plants can also contribute significantly to the country’s economy. In the
mean time, a large number of industries (400 herbal factories) have been established
in the country for producing Ayurvedic & Unani medicines. It has been estimated that
Bangladesh has a market of about 300 core taka worth herbal product annually. A
recent study on Medicinal Plants Marketing in Bangladesh sponsored by SEDF &
Interco operation (IC) conducted in October 2003, reviewed the current status &

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estimated the quantity & value of medicinal plants used as raw materials both in
organized sector (large companies, small companies),unorganized sector (herbal
doctors/practitioners) & spent annually on approximately 17500 tones medicinal
plants materials accounting approximately Tk.81 Cr. Currently , the export of
medicinal plant is in a rising state. United Arab Emirates is the top export market for
Bangladesh followed by Pakistan & United Kingdom. (20).
Medicinal plants can serve as templates or raw materials for further modification and
synthetic remodeling; those with high potential can minimize dependency on foreign
raw materials and be imported after meeting local demand. But this is a matter of
great concern that this potential sector is not gaining much attention to flourish.
Recently, some research institutes like, Bangladesh Centre for Scientific and
Industrial Research (BCSIR), some universities namely University of Dhaka,
Jahangirnagar university and some non-governmental organizations are collaborating
to boost the country's research, production and conservation of medicinal plants.
Bangladesh Agricultural research institute is trying to establish a medicinal plant gene
bank. Traditional records & ecological diversity indicate that Bangladeshi plants
represent an exciting resource for possible lead structures in drug design.

1.4. Research on medicinal plants: Bioactivity guided approach

Bioactivity guided phytochemical investigation of medicinal plants may yield such
chemical constituents having no remarkable therapeutic interest. The crude drug
containing several constituents may be found to be ineffective in case of therapy for
which it was used traditionally. For example, Vinca rosea once used traditionally as
antidiabetic drug was found to contain anticancer principle vinca alkaloid in a high
yield rather than hypoglycemic principle. Rauwolfia serpentina which was
traditionally used for a variety of illnesses, revealed the presence of an
antihypertensive and tranquilizing agent reserpine. It is evident from the above
discussion that the search of plant constituents having therapeutic interest requires
bioactivity studies with the crude extracts prior to phytochemical analysis. Without
prior pharmacological studies, the phytochemical studies alone can provide the
chemical constituents of the plant, but these may or may not have therapeutic value.
So in medicinal plants research, bioactivity guided phytochemical approach might be
a rational approach.

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Bioactivity guided phytochemical approach has three phages of investigation.

 First, biological activity is detected in crude material by bioassay system
which permits the identification of active fractions and discarding the inactive
ones.
 Second, the crude material is fractionated by the most appropriate chemical
procedures, all fractions are tested and active fractions are further fractionated
and so on, until pure compounds are obtained.
 Third, the chemical structures of pure compounds are determined.

1.5. Objectives

Plants provide a bank of rich, complex, and highly varied structure and biologically
important compound that are of medicinal value.Grewia nervosa (Malvaceae) is also
a plant which has many ethno-pharmacological uses. Such as ---- Tribal people
(Tripura) use this plant to treat jaundice [21) and many local people in Chittagong hill
tracts use this plant for wound healing purpose. A few chemical and pharmacological
works have been carried out in this plant. There remains a possibility that the extracts
of different parts of the plant may contain some bioactive compounds as they have
various ethno-pharmacological uses. The work described in this dissertation is an
attempt to evaluate the possible pharmacological activities of the crude extract to

 Rationalize the traditional uses of the selected plants

 Explore the possible newer medicinal activities of the same plant
 Strengthen the further research of plant extract for pre-clinical studies to
determine pharmacological effects.
 To ascertain the safety and efficacy of extracts on the different types of animal
models [22]
 Different analogues and more potent synthetic drugs can be prepared by
structure activity relationship (SAR) and molecular modification.

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1.6.The study protocol

Our present study was designed to observe pharmacological activities of the crude
extract & their different partitioning fractions 0f leaf part of plant Grewia nervosa.

It consisted of the following steps:

1.6.1. Phytochemical investigation

 Extraction of the plant Grewia nervosa with methanol for fifteen days.
 Filtration of the crude methanolic extract by using the Markin cotton
cloth and subsequently through the filter paper and solvent evaporation
with the help of rotary evaporator.
 Fractionation of methanolic crude extract with n-Hexane, pet ether,
chloroform and dichloromethane by modified Kupchan partition
method.

1.6.2. Phytocemical screening

1.6.3. Pharmacological investigation

In vivo test :

 Evaluation of peripheral analgesic effect of crude extract & different

fractions of Grewia nervosa by acetic acid induced writhing method.
 Evaluation of central analgesic effect of crude extract & different
fractions of Grewia nervosa by Tail immersion method in swiss albino
mice.
 Investigation of CNS depressant activity of methanolic crude extract &
different fractions in swiss albino mice by-
 Hole-board test
 Elevated plus maze test
 Evaluation of hypoglycemic activity of crude extract of by Glucose
tolerance test in swiss albino mice.
 Evaluation of antidiarrheal activity of crude extract & different fraction of
Grewia nervosa by castor oil induced diarrhea method in swiss albino
mice.

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 Screening of anti-pyretic activity of methanolic extract of G.nervosa by

Yeast induced hyperthermia method in swiss albino mice.
In vitro test
 Evaluation of thrombolytic activity of crude extract of Grewia nervosa by
in vitro clot lysis method on human blood.
 Screening of in vitro anti arthritic activity of crude extract of Grewia
nervosa by Bovine serum albumin (BSA) denaturation method.
 Evaluation of antibacterial assay of crude extract by disc diffusion method.
 Brine shrimp lethality bioassay and determination of LC50 of crude extract
and
different fractions of Grewia nervosa..
 Determination of anthelmintic activity of methanolic crude extract of
Grewia nervosa .

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