Download HerbalDrugs_Thailand

Development of Pharmaceutical Products from Medicinal Plants
Section 1 General Information
a. Title of innovative experience on the Use of Phyto-Pharmaceuticals from Medicinal
Plants.
Role of Science and Technology in Promoting the Use of Herbal Drugs: An Example
of Andrographis paniculata.
b. Keywords : up to 10 key words that characterize the innovative experience
Andrographis paniculata, Cultivation, Therapeutic efficacy, Side effects, Diterpenoids,
Variation, Product Stability, Appropriate Technology.
c. Name and address of implementing institution (include fax, phone, email, web)
Chulabhorn Research Institute (CRI)
Vipavadee Rangsit Highway, Laksi, Bangkok 10210, Thailand.
Telefax
: (66-2)-574-0616 or 575-1497
Telephone : (66-2)-574-0615 or 574-0622-33 ext 1610
E-mail
: [email protected]
Web
: http://www.cri.or.th
d. Name and address of the head of the institution.
Professor Dr. HRH Princess Chulabhorn
President, Chulabhorn Research Institute
Vipavadee Rangsit Highway, Laksi, Bangkok 10210, Thailand.
e. Name and address of the person responsible for providing detailed information on
the innovative experience.
Assoc. Prof. Dr. Jutamaad Satayavivad
Office of Academic Affairs
Chulabhorn Research Institute
Vipavadee Rangsit Highway, Laksi, Bangkok 10210, Thailand.
f. Individuals involved in the innovative experience, with a brief indication of their role
in developing the experience.
1. Professor Dr. HRH Princess Chulabhorn
To set the policy of the Institute to conduct a project which can apply
science and technology for the benefit of the people at grass roots level.
2. Assoc. Prof. Dr. Mathuros Ruchirawat, Vice President for Research
To carry out the policy by co-ordinating and implementing the project entitled
“Green Health Technology for Women’s Empowerment and Sustainable
Development” which received support from UNDP (UNDP/RAS/97/001)
One of the activities of this project involves the cultivation, processing and
marketing of herbal medicine to generate additional income for women in poor
rural areas.
3. Professor Dr. Somsak Ruchirawat, Associate Vice President for Research
To be responsible for the phytochemical studies of the selected medicinal
plant products.
4. Assoc. Prof. Dr. Jutamaad Satayavivad, Associate Vice President for Academic Affairs
To be responsible for field and laboratory studies of the medicinal plant
products.
g. Implementation period and total cost (expressed in US dollars) of the project
(list contributions received from government, private sector and foreign sources)
Feasibility Study : 1999
Project Period
: (2000-2002)
Total cost
: US$ 2,265,382
UNDP
: US$ 76,500
CRI
: US$ 2,188,882
-2-
Section 2 Non-technical summary of the main features of the project. (highlight
the innovative aspects and the main results involved)
During January 2000-December 2002, the Chulabhorn Research Institute (CRI)
received funding from United Nations Development Programme (UNDP) to implement
the project entitled “Green Health Technology for Women’s Empowerment and
Sustainable Development”. The Use of Medicinal Plants (Curcuma longa and
Andrographis paniculata) for simple illness, and cultivation. as a source of additional
income is one of the components of this project. However, in this case study only A.
paniculata will be described to illustrate the integration of field and laboratory activities
which benefits all parties involved.
It is generally accepted that both in the case of a modern drug or an herbal drug,
its therapeutic efficacy and safety must be proven. Variations of active ingredients of
herbal drugs are also well known, and account for therapeutic ineffectiveness and side
effects. To promote the continuous use of herbal drugs, scientific intervention is
necessary at all steps, starting from good agricultural practice to good manufacturing
practice. The project undertaken by CRI covering both field and laboratory activities has
been conducted in parallel. CRI’s scientists have successfully isolated, three major
active chemicals (andrographolide (AP1), 14-deoxy-11,12-didehydroandrographolide
(AP3) and neoandrographolide (AP4)) from this plant, and used them as reference
compounds, to study their pharmacological and toxicological activity. Furthermore, a
simple and rapid method using High Performance Liquid Chromatography (HPLC) to
analyse three major active chemicals simultaneously has been successfully developed.
This analytical technique has a wide application, for example, it has been used for
(a) Quality control of the finished A. paniculata powdered and extracted products.
(b) Study the stability of the A. paniculata products.
(c) The selection of appropriate cultivation period to obtain the maximum contents of
the desired active compounds.
One of the few side effects observed in some patients using A. paniculata
products is lowering of blood pressure. We are able to identify that AP3 exhibits an
antihypertensive effect, and its content in the products increased upon storage;
therefore, we suggested that to avoid this side effect, the A. paniculata crude powdered
products should be used within one year. At present, attempts have been made to find
the appropriate cultivation condition that results in high content of AP 3, and study its
potentiality as an antihypertensive drug.
In summary this project has achieved the followings:
1. The appropriate conditions for cultivation and harvesting of A. paniculata have
been identified.
2. Isolation of the pure active diterpenoids has been achieved.
3. A rapid and simple HPLC method has been developed to analyse three
bioactive diterpenoids simultaneously.
4. Bioactive compounds causing lowering of blood pressure have been identified.
5. Shelf-life of A. paniculata crude powdered drug has been estimated.
6. A preliminary study using water extract, partially purified extracts and pure
diterpenoids revealed that these test compounds exhibit antimalarial activity,
antiplalelet aggregation, and lowering of blood pressure.
7. Supplementary income generation for women participating in this project has
been achieved.
-3-
Section 3 Background and Justification
a. Brief description of the situation before the innovative experience
Since 1980, the office of Primary Health Care System, Ministry of Public Health,
had proposed herbal drugs as Essential Drugs which could be used as one of the
strategies to achieve “Health for All by the year 2000”. A medicinal plants
demonstration garden had been set up at the health station. However, a large
variety of medicinal plants continues to be gathered from their wild sources. In the
last decade there has been a resurgence of interest in herbal medicines in western
societies. With the rapid increase of market demand, cultivation of medicinal plants
is obviously needed. The first volume of Thai Herbal Pharmacopoeia was
introduced in 1995. This Pharmacopoeia compiles specifications and related data
concerned with the quality of herbal drugs. The National Essential Drug List
Committee of the Ministry of Public Health released “The National List of Essential
Drugs A.D. 1999-List of Herbal Medicinal Products”, Curcuma longa and
Andrographis paniculata are included in this list.
The ultimate goal of the Chulabhorn Research Institute is to use science and
technology to improve “Quality of Life”
After the successful organization of the Princess Chulabhorn Science Congress
in 1999, His Majesty King Bhumibhol advised HRH Princess Chulabhorn, the
President of Chulabhorn Research Institute to develop a project which will benefit
the people at the grass roots level.
HRH Princess Chulabhorn held an executive meeting to formulate the project
according to the advice of her father. During this period, CRI was in the process of
preparing the project proposal to seek support from UNDP. Therefore, it was agreed
that the Institute would use science and technology to improve the quality of life of
the people not only through medicinal plant utilization but also through cultivation as
a source of supplementary income for poor families in rural areas.
b. Justification and context of the innovation experience
Because the government set the policy to promote medicinal plants as a
component of essential drugs for the primary health care system of the country,
cultivation of medicinal plants has been necessary, especially for those plants that
are listed in the essential drug list. From the feasibility study, two medicinal plants
(Curcuma longa and Andrographis paniculata) had been selected for cultivation. C.
longa was selected because of its wide uses in food, cosmetics, and health. The
variety that gave the highest yields of volatile oils and curcuminoids was selected for
cultivation. The rhizomes will be sold as raw materials to the local drug industry. A.
paniculata was selected because it has been used widely by the lay public to treat
the common cold. From personal communications with Thai traditional doctors, it
was found that this medicinal plant caused a drop in blood pressure in some
patients. As the scientific literature shows, this medicinal plant has a wide spectrum
of pharmacological activities, for example, antiviral (Anti-HIV), common cold,
antihepatoxicity, filaricidal activity, antidiarrheal activity, antimalarial activity,
immunostimulant and cardiovascular activity. It has been documented that the total
contents of lactones in A. paniculata from different sources are similar. However, the
contents of each diterpenoid lactone vary according to the time of harvest and the
season of the year. Because the Chulabhorn Research Institute is a non-profit
organization and cannot manufacture medicinal plant products for sale, our project is
aimed at assisting local communities to produce quality raw materials for
supplementing income and at assisting local manufacturers to produce high quality
-4-
products by applying science and technology. At present, more than twenty A.
paniculata preparations are available in the local marketplace. Among these, very
few are manufactured by well known factories with good manufacturing practice.
To avoid variations of the bioactive chemicals present in this medicinal plant,
recently, several derivatives of pure compounds present in this plant have also been
synthesized and are in the process of patenting. It is anticipated that these products
will be too costly for poor people in developing countries. Appropriate technology
should, therefore, be applied to obtain a low cost and good quality pharmaceutical
product.
c. Main issues involved
In general, the derivation of well-defined and effective pharmaceutical products
from medicinal plants requires at least 2 steps, good agricultural practice and good
manufacturing practice.
Cultivation of selected medicinal plants should follow good agricultural practice
(GAP). Pesticides were not used in this project. Appropriate harvesting time must be
identified.
It is well documented that there is a wide variation of the active compounds of this
medicinal plant depending on the harvesting time and season of the year. To solve this
problem, a standardized testing procedure is required. To achieve this goal, biomarkers
for the biologically active compounds are needed. Ideally, a simple and rapid analytical
technique should be developed that can be used in the quality control of both the raw
materials and finished products, and as means to study the shelf-life of products to
ensure their safety and therapeutic effectiveness.
-5-
Section 4 Description
a. Detailed description of the experience.
This project is aimed at applying sophisticated science and technology to benefit
the rural community. Two major complementary activities were conducted in parallel,
namely, practical activities at the rural community level, and scientific and technical
activities at the CRI laboratories.
1. Cultivation and processing of medicinal plants.
The cultivation area was improved by prior ploughing and legume planting; no
chemical fertilizers were used. Cultivation of A. paniculata (in Thai “Fah Thalai Jone”)
was starting from seed. Germination was done in cups containing a 1:1 mixture of soil
with leaf mold, manure with peat moss, or peat moss alone. One to 3 seeds were
placed in each cup in germination trays. Forty-five days after initial seeding, seedlings
were transplanted into the cultivation area. After three months, robust leafy plant growth
was seen as shown in figure1. At the fourth month about 50% were blooming and the
plants were ready for harvest. For the first two crops, the leaves were gathered by hand,
air dried, then oven dried at low temperature (approx. 50°C) to a constant weight,
weighed and packed; however, since this was time consuming, an alternative method
was introduced, involving hand cutting of the whole plant at ground level. Either fresh or
dried leaves were removed from the whole plants using a rotary agitator to dislodge the
leaves and a forced air current to blow the leaves into a collecting bin. This equipment
was fabricated and operated at the project site. Part of the processed medicinal plant
materials was sold to traditional medicine outlet for profit. The other part of the product
from the cultivation area was used by the CRI laboratories. Good quality dried leaves of
A. paniculata could be sold for US$ 5/kg (US$ 1 = 40 Baht), while purified active
ingredients, andrographolide and its derivative cost as much as 4.9 million baht/kg from
a chemical supply house. CRI calculations based on the estimated content of these
pure compounds in A. paniculata leaves predict that it will be more profitable in the
future to purify the active bioactive chemicals for sale.
2. Laboratory activities.
The scientific activities carried out at CRI’s laboratories included,
- Initial soil and water analyses to determine optimal cultivation site characteristics.
- Analysis of soil and water by Atomic Absorption Spectrometric methods to
ensure that levels of heavy metal contaminants in soil and water did not exceed
safe limits.
- Using Gas Chromatography to analyse for contaminating pesticides in soil and
water.
- Determining optimum conditions for seed germination, planting time, growth
period and harvest time to maximize the yield of bioactive chemicals.
- Refining the techniques for isolation, purification and structural characterization
of diterpenoids (andrographolide and its derivatives), which are the bioactive
compounds in A. paniculata, using High Performance Liquid Chromatography,
Nuclear Magnetic Resonance, and Mass Spectrometry. Purified active
compounds were used as standards in HPLC analysis.
- Studies of the pharmacological properties of partially purified extracts and pure
compounds isolated from A. paniculata.
- Development of a rapid and simple HPLC method to quantify three major active
compounds (Andrographolide, 14-deoxy-11,12-didehydroandrographolide, Neoandrographolide) simultaneously.
-6-
Figure 1: Cultivation of Andrographis paniculata
(A)
(B)
(C)
(D)
(A) Seed germination
(B) 45-60 days after initial seeding
(B) 3 months after seeding
(D) Flowering plants
-7-
b. Steps taken in the planning and implementation process.
Before starting the project, a feasibility study was conducted to select the project
site and the women participating in the project. Tubtim Siam Village 5, Srakaew
Province, which is near the Cambodian border was selected as the project site. The
cultivated medicinal plants were selected because of their well-known therapeutic
efficacy, safety, and economic potential. Project committee was appointed and key
personnel of the project were identified. Training for cultivation with good agricultural
practice (GAP) was initially done by Department of Pharmaceutical Botany, Faculty
of Pharmacy, Mahidol University. Later the monitoring and supervision were
conducted by a team from CRI. The first lot of harvested leaves and stems were
sent to CRI’s chemistry laboratory for isolation and identification of the major
diterpenoid compounds. The pure bioactive compounds were used by the
pharmacological group to develop the HPLC analytical technique and to study the
pharmacological activities as showed in figure 2.
Figure 2: Steps Taken in the Implementation Process
Role of Science and Technology in Promoting the Use of Herbal Drug:
An Example of Andrographis paniculata
Project Committee
(Appointment of
Key Personnel)
Coordinating
Group
Phytochemical
Group
-
Extraction
Separation
Purification
Isolation
Structure Elucidation
Extracts
Selection of the Project Site and Women
Participating in the Project
Supervision by a
Team of Experts
from Mahidol
University
Cultivation of
A. paniculata
with GAP
Monitoring and
Supervision by a
Team of CRI
Harvesting and
Processing
Product /
Marketing
Pure Bioactive
Compounds
Local
Herbal Drug
Manufacturers
Pharmacological
Group
Development of a rapid
and simple HPLC
analytical method
Pharmacological studies i.e.
Antimalarial activity,
Antihypertensive activity,
Antiplatelet aggregation
-8-
c. Technical data including diagrams, charts, tables, photographs
1.Cultivation:
A. paniculata harvested just before flowering contained the three active
triterpenoids in leaves at higher concentration than in the stems as shown in table 1.
Table1: Concentrations of andrographolide (AP1), 14-deoxy-11,12-didehydroandrographolide (AP3) and neoandrographolide (AP4) in leaves and
stems of A. paniculata. Values represent the mean of three replicates
with standard error of means.
Sample
Contents (mg/g DW)
AP1
AP3
AP4
Leaves
17.450.16
17.380.11
6.140.07
Stems
8.370.12
2.640.06
0.370.02
2. Extraction and isolation of active ingredients of Andrographis paniculata
The extraction and isolation of the active ingredients of A. paniculata are
summarized in figure 3.
Three major bioactive compounds are identified:
andrographolide (AP1), 14-deoxy-11,12-didehydroandrographolide (AP3) and neoandropholide (AP4).
3 Study of the effect of diterpenoids on the cardiovascular system
The three major bioactive compounds AP1, AP3 and AP4 were studied for their
effects on the cardiovascular system. It was found that AP 3 could significantly
decrease the beating rate of the isolated right atrium of rats when compared with a
control group. Furthermore, it was found that AP3 reduced vasocontraction induced
by norepinephrine in the isolated descending aorta of rats more strongly than AP 1,
while AP4 did not have this effect.
4 Variation of bioactive compounds in the herbal products
Concentrations of the three tripenoids in the recommended daily dose were
showed in figure 4. Twenty A. paniculata preparations were purchased from local
drug stores. The contents of AP3 in some preparations were rather high (Products
1,5,8,13,14 and 15), which may account for the observed effects on the blood
pressure.
5 Stability of the three tripenoids
Dried powder of A. paniculata harvested at different times in 2001 (S1-18 July;
S2-25 July; S3-26 July, S4-8 August) were kept in plastic bags and stored at room
temperature (approx. 28°-30°C). Samples (300 each, were extracted and analyzed
for the contents of three tripenoids (AP1, AP3 and AP4) at 0,3,6,11,12 and 15 months
using the HPLC analytic method developed by CRI laboratories. The results are
shown in figure 5. It should be noted that the contents of AP 3 in all tested samples
increased with storage time while active compounds (AP 1 and AP4) decreased with
time. These results suggest that the stability of these active compounds should be
taken into consideration when this herb is stored for a period of time to ensure the
efficacy and safety of the herbal products.
-9-
Figure 3: The Extraction and Isolation of Active Ingredients
of Andrographis paniculata.
Dried Powdered Leaves (3.50 kgs)
- macerated with n-hexane
(~8.0 Lit. for 2 days, 3 times)
- filtered, evaporated
70
Hexane
Extract
Andrographolide
Marc
Diterpenoid contents
(mg/g DW)
14-deoxy-11,12-didehydroandrographolide
Green-brown
Neoandrographolide
60
(10.88 grams)
- macerated with CH2Cl2
(~6.0 Lit. for 2 days, 3 times)
- filtered, evaporated
50
Marc
40
CH2Cl2 Extract
Green-black
(112.89 grams)
- macerated with MeOH
(~5.0 Lit. for 2 days, 5 times)
- filtered, evaporated
30
20
MeOH
Extract
Marc
Green-black
10
(395.17
grams)
Andrographolide (~ 6 grams)
Neoandrographolide
14-deoxy-11,12-didehydroandrographolide
0
01
02
03
04
05
06
07
08
09
10
11
Products
- 10 -
12
13
14
15
16
17
Compounds …....
18
19
20
AP1
AP3
Figure 4:
Variation of andrographolide (AP1), 14-deoxy-11,12-didehydroandrographolide (AP3) and neoandrographolide (AP4) contents in
A. paniculata products from different suppliers in Thailand. Values
APof1 three replicates with standard error of means.
represent the mean
- 11 -
S1
S2
S3
S4
35
Diterpenoid contents
(mg/g DW)
30
25
20
15
10
5
0
0
3
6
11
12
15
Storage time (month)
AP3
40
35
Diterpenoid contents
(mg/g DW)
30
25
20
15
S1
S2
S3
S4
10
5
0
0
3
6
11
12
15
Storage time (month)
AP4
S1
5
S2
S3
Diterpenoid contents
(mg/g DW)
4
S4
3
2
1
0
0
3
6
11
12
15
Storage time (month)
Figure 5: Changes
in
andrographolide
(AP1),
14-deoxy-11,12-didehydroandrographolide (AP3) and neoandrographolide (AP4) contents in dry herb
of A. paniculata stored at room temperature up to 15 months (values
represent the mean of three replicates with standard error of means).
Samples S1, S2, S3, S4 harvested at 18 July, 25 July, 26 July and 8 August
2001, respectively.
- 12 -
Section 5 Partnerships
a. Public/community/stockholders and private sector involvements
This project was carried out with close collaboration among community,
academic and industry groups. Initially the linkages with private sectors emphasizing
selling the raw materials was not satisfactory because each party wanted the most
benefit for itself. However, in the second step, involving the mutual interest in
developing the herbal drugs for poultry, the linkage between the private sector and
the institute was strengthened because both parties wanted to assist poor farmers.
b. Other regional and international collaborators (if any)
The activity of this project was partly funded by UNDP through the project “Green
Health Technology for Women’s Empowerment and Sustainable Development”. The
initial results of this activity were presented in the Chennai Follow-up Meeting in
November 23-25, 2000. The theme of the meeting was “Utilizing Science and
Technology for Women’s Economic Empowerment: Progress and New challenges”
A video tape of this activity at the end of the project has been prepared and can be
distributed upon request.
- 13 -
Section 6 : Replicability
a. Relevance of the innovative experience to other regions.
Medicinal plants have been used since ancient time in various parts of the world.
The proper application of medicinal plants has been practiced by knowledgeable
traditional doctors. In several countries, the popularity of medicinal plants has
declined in parallel with mass deforestation; therefore, the revitalization of the use of
medicinal plants demands both a stable and adequate supply of medicinal plants
and accurate knowledge of traditional uses. Cultivation of selected medicinal plants
and their qualitative control are essential for effective use. Promotion of the use of
herbal drugs, therefore, relies heavily on the availability of good quality raw
materials. The knowledge gained from this project can be applied to other regions.
However, such projects must be appropriately supported and have well-equipped
laboratories.
b. Policy implication
This intensive study was conducted as a consequence of a clear policy of the
government to promote the utilization of herbal drugs and the policy of the Institute
to use science and technology to improve the quality of life of the people. The
success of similar projects will need a clear policy of the organization to use the
experience learnt from this project in the development of pharmaceutical products
from the selected medicinal plants.
c. Changes in legislation brought about thanks to the innovative experience
At present, there are no changes in legislation. However, recommendation for
the quality control of the products from this medicinal plant can be suggested. The
quality control of A. paniculata was set by the Ministry of Public Health. It is stated
that the total lactone content calculated as andrographolide must not be less than
6.0%. A simple method for analysis of total lactone is described. The results of this
study suggest that the total lactone content is not an appropriate marker, since the
total lactone content of two preparations may not be significantly different, but the
content of each bioactive diterpenoid may be markedly different which will affect the
therapeutic effectiveness and also side effects of A. paniculata preparations.
Therefore, the result of this study will be submitted to the Ministry of Public Health
for consideration in modifying the standard of this medicinal plant for a specific
therapeutic indication. In addition, A. paniculata preparations used for sore throat
should have standards relating to shelf-life of the preparation. (For example,
preparations should be used within one year in order to avoid side effects related to
the lowering of blood pressure.)
- 14 -
Section 7 : Impact
a. Quantitative and/or qualitative impact of the experience after Implementation
(indicator of success)
The completion of the first phase of this project coincides with the urgent need of
the country to develop herbal drugs for poultry. Antibiotic residues in chicken have
posed a serious problem for the economy of the country because contaminated
products have been rejected for export purposes. Therefore, use of most antibiotics
has been banned. Alternative drugs for poultry therefore represent an urgent need.
The study on A. paniculata for controlling respiratory disease in poultry, therefore,
came just at the right time. Knowledge of appropriate cultivation of A. paniculata and
the development of the analytical technique to quantify the three bioactive
compounds simultaneously is very useful for further development of this herbal drug
for use in poultry. Furthermore, the identification of AP3 as the most active chemical
for lowing blood pressure is important, especially when it was discovered that this
compound increases during storage. This information is useful for avoiding side
effects. Furthermore, isolation of this compound for its use as an hypertensive drug
may be beneficial and further study in this aspect is ongoing.
b. Sustainability of the innovative experience (economic, environmental, social)
Demand for herbal drugs is increasing and maintaining an adequate supply from
wild sources is not sustainable. Cultivation of herbal drugs can be sustainable,
economical and environmentally sound. Germination of A. paniculata seeds for a
selected period of cultivation is not easy. This herbal plant naturally grows easily
during rainy season, but to prevent over supply of A. paniculata, it should be
cultivated throughout the year according to demand. However, as the result, the
concentration of active ingredients in the leaves will vary. Such variation can be
detected and standardized by using the simple and rapid analytical method that has
been developed by our institute applying standard chemicals isolated and purified by
phytochemists of the Institute. This innovation makes production of the herbal drug
sustainable and economical. This analytical method will be published in an
international journal and will benefit all who are working with this medicinal plant.
c. Special circumstances which made the experience successful
The success of this experience depended on the effort of a number of personnel
of the Institute in carrying out the policy which was set according to the wishes of His
Majesty the King.
This integrative project needed the skills of a wide variety of experts including
economists, botanists, agriculturists, pharmacists, phytochemists, pharmacologists,
toxicologists. These personnel have to work together collaboratively. This is a new
project for the Institute which involves villagers at the project site (which is a
community development project), the private sector, and scientists. Good
cooperation among all partners was essential, and many obstacles were solved
through the efforts of the involved parties because this was a project initiated by the
Royal Family.
- 15 -
Section 8 : Lessons Learnt
a. Obstacles faced and steps taken to overcome them
The objective of the project is to use science and technology to improve the
quality of life of the poor and to discover how to transfer the knowledge from
laboratory work for practical uses to the people who not only have no background in
science but also poor literacy. The women participating in this project had low
educational background and income. The first step undertaken was to explain the
benefits of producing good quality raw materials and how to make them. The
following are examples of lessons learnt.
1. Good Agricultural Practice and Marketing
To produce raw materials for herbal drugs, good agricultural practice is very
important. No pesticides are allowed; therefore, it is labor intensive. Weeds have to
be picked out by hand. During the harvesting period, leaves were also picked by
hand and air or oven dried at low temperatures during rainy season. Raw materials
(with only dry leaves) were intended to be sold at 200 baht/kilogram (or approx. US$
5/kg.). This price was found to be too high and could not compete with lower grade
raw materials. To solve this problem, studies of seed germination at various times of
the year were conducted so that A. paniculata could be cultivated according to the
market demand. The 4 largest herbal drug manufacturers were invited to inspect the
raw materials and to see video tape showing good agricultural practice. They were
asked to order raw materials about 4 months in advance. However, this approach
was not fully successful because of the higher price of the raw materials. The final
step was to send raw materials to the Institute’s laboratory for isolation of the pure
bioactive compounds.
2. Application of Science and Technology to make value added products
The lesson learnt from good agricultural practice is that the price of the raw
material is higher than other producers, and most of the local manufacturers still can
not afford to buy the good quality materials without raising the price of the products.
The institute is well equipped with modern scientific equipment and competent
scientists. Therefore, the unsold A. paniculata were used as a raw materials for
isolation of pure biological active compound. The catalogue price of andrographolide
is US$ 61.50/500 mg. Several pure compounds were isolated and being used as
standard drugs for the institute’s internal use, and also distributing to the other
government agencies upon request.
The next step to overcome this problem is to develop the product for use in
poultry. Recently, the government imposed strict control on the uses of antibiotics in
poultry. From traditional knowledge, it was known that A. paniculata could be used
to treat respiratory disease in poultry. The demand of herbal drugs for poultry is very
high and there is a big market for A. paniculata as compared with uses for treating
diseases in human. Therefore, the development of A. paniculata pharmaceutical
product for poultry is on going with the collaboration of Tanaosri Farm in Rajaburi
Province, Thailand.
- 16 -
3. Income generation for the Institute
CRI is a non-profit organization; therefore, manufacturing of herbal drugs cannot
be undertaken, nor can the selling of pure standard chemicals. Seeking private
partnership should be explored. At present, the most appropriate approach is to
study the pharmacological potential of the available pure chemicals. So far one
chemical has been found to exhibit antihypertensive and antiplatelet aggregating in
animal models.
b. Preparing public perception for acceptance of the innovation
A representative of CRI was invited to participate in a closed meeting of
academia, industry and government agencies about using medicinal plants in
animals. A. paniculata was selected as a potential medicinal plant. Plans for
cultivation, quality control of the products, and side effects have been made. The
representative of the Institute had an opportunity to share the experience and
express willingness to cooperate with the private sector.
- 17 -
Section 9 : Future Plans
a. Plans for further improvements and expansion of the project
At present, cultivation of A. paniculata from seeds in order to study the contents
of each active compounds at different time intervals after seeding has been
conducted. If it is successful, it will facilitate the isolation of pure active compounds
which can be sold as standard chemicals.
Since A. paniculata products available in the market, at present, are mostly in the
form of crude dried powder from leaves and stems. Modification of drug
preparations in the form of capsule containing standardized extract and spry dry will
improve the quality of the medicinal plant products, especially from microbial
contamination. Furthermore, large scale isolation of pure biological chemicals will
be conducted since these pure chemicals are very useful in the quality control of
herbal drugs. Further studies on the pharmacological and toxicological effects which
will lead to the development of modern drugs should be conducted. At present,
even though the analogues of several pure chemicals have been synthesized and
were patented, these products will be costly; therefore, it is anticipated that herbal
drugs will still be needed in the developing countries. Since the yield of
andrographolide is quite high and the process of extraction, separation, isolation
purification has been known, the production in the form of standardized extracts from
natural source will be inexpensive; furthermore, it will generate additional income for
poor farmers in the developing countries.
b. Plans for collaboration and sharing results with other organizations/countries
A video type of the project has been made and it will be distributed upon request.
The scientific results will be published in an international journal (J. Phytochemical
Analysis). The Institute will give service with minimal charge for quality control of
local products of A. paniculata.
- 18 -
REFERENCES
Burgos, RA, Imilan, M, Sanchez, NS, and Hancke, JL. 2000. Andrographis paniculata
(Nees) Selectively blocks voltage-operated calcium channels in rat vas deferens.
J Ethnopharmacol 71: 115-21.
Burgos, RA, Aguila, MJ, Santiesteban, T, Sanchez, NS, and Hancke JL. 2001
Andrographis paniculata (Nees) induces relaxation of uterus by blocking voltage
operated calcium channels and inhibits Ca+2 influx. Phytother Res 15: 235-9
Caceres, DD, Hancke, JL, Burgos, RA, Sandberg, F, and Wikman, GK. 1999. Use of
visual analogue scale measurements (VAS) to assess the effectiveness of
standardized Andrographis paniculata extract SHA-10 in reducing the symptoms
of common cold. A randomized double blind-placebo study. Phytomedicine 6:
217-223.
Calabrese, C, Berman, SH, Babish, JG, Ma, X, Shinto, L, Dorr, M, Wells, K,
Wenner, CA, and Standish, LJ. 2000. A phase I trial of andrographolide in HIV
positive patients and normal volunteers. Phytother Res 14: 333-338.
Chauhan, SK, Singh, BP, Kimothi, GP, and Agrawal, S. 1999. Determination of
andrographolide in Andrographis paniculata by High Performance Thin Layer
Chromatography. Indian Drugs 36: 130-132.
Cheung, HY, Cheung, CS, and Kong, CK. 2001. Determination of bioactive diterpenoids
from Andrographis paniculata by micella electrokinetic chromatography. J
Chromatogr A 930: 171-176.
Chiou, WF, Lin JJ, and Chen, CF. 1998 Andrographolide suppresses the expression of
inducible nitric oxide synthase in macrophage and restores the vasoconstriction
in rat aorta treated with lipopolysacharide. Br J Pharmcol 125: 327-34.
Choudhury, BR, Haque, SJ, and Poddar, MK. 1987. In vivo and in vitro effects of
kalmegh (Andrographis paniculata) extract and andrographolide on hepatic
microsomal drug metabolizing enzymes. Planta Med 53: 135-40.
Clander, R, Srivastava, V, Tandon, J, and Kapoor, NK. 1995. Antihepatotoxic activity of
diterpenes of Andrographis paniculata (Kal-Megh) against Plasmodium berghei
induced hepatic damage in Mastomys natalensis. Int J Pharmacog 33: 135-138.
Dechatiwongse Na Ayudhya, T, Techadamrongsin, Y, and Jirawattanapong, W. 1993.
In: Chemical specification of Thai herbal drugs. Vol 1, Department of Medicinal
Sciences, Ministry of Public Health, Bangkok. P. 25-41.
Du, Q, Jerz, G, and Winterhalter, P. 2003. Separation of andrographolide and
neoandrographolide from the leaves of Andrographis paniculata using highspeed counter-current chromatography. J Chromatogr A 984: 147-151.
Dutta, A. and Sukul, NC. 1982. Filaricidal properties of a wild herb, Andrographis
paniculata (Nees) J Helminthol 56: 81-4.
Gupta, S, Yadava, JNS, and Tandon, JS. 1993. Antisecretory (antidiarrhoeal) activity of
Indian medicinal plants against Escherichia coli enterotoxin-induced secretion in
rabbit and guinea pig ileal loop model. Int J Pharmacog 31: 198-204.
Handa SS and Sharma A. 1990. Hepatoprotective activity of andrographolide against
galactosamine and paracetamol intoxication in rats. Indian J Med Res (B) 92:
284-292.
Jain, DC, Gupta, MM, Saxean, S and Kumar, S. 2000. LC analysis of hepatoprotective
diterpenoids from Andrographis paniculata. J Pharm Biomed Anal 22: 705-709.
- 19 -
Jewvachdamrongkul, Y, Jirawattanapong, W, and Dechatiwongse, T. 1990. Modified
method for determination of total lactones in Andrographis herb. Bull Dept Med
Sci 32: 53-61.
Kapil, A, Koul, IB, Banerjee, SK, and Gupta, BD. 1993. Antihepatotoxic effects of major
diterpenoids constituents of Andrographis paniculata. Biochem Pharmacol 46:
182-185.
Laorpaksa, A, Amnuoypol, S, and Jongbunprasert, V. 1988. Preliminary study on
antibacterial action of Thai medicinal plants for respiration tract infection (I).
Thai J Pharm Sci 13: 23-36.
Li, W and Fitzloff, JF. 2002. Determination of andrographolide in commercial
andrographis (Andrographis paniculata) products using HPLC with evaporative
light scattering detection. J Liq Chromatogr Related Technol 25: 1335-1343.
Lomlim, L, Jirayupong, N and Plubrukarn, A. 2003. Heat-accelerated degradation of
solid-state andrographolide. Chem Pharm Bull 51: 24-26.
Matsuda, T, Kuroyanagi, M, Sugiyama, S, Umehara, K, Ueno, A, and Nishi, K. 1994.
Cell differentiation-induced diterpenes from Andrographis paniculata. Chem
Pharm Bull 42: 1216-1225.
Ministry of Public Health. 1995. Fa Tha Lai. In: Thai Herbal Pharmacopoeia. Vol 1,
Department of Medicinal Science. Prachachon Co. Ltd., Thailand, 24-31.
Misra, P, Pal, NL, Guru, PY, Katiyar, JC, Srivastava, V, and Tandon, JS. 1992.
Antimalarial activity of Andrographis paniculata (Kalmegh) against Plasmodium
berghei NK 65 in Mastomys natatensis. Int J Pharmacog 30: 263-274.
Najila, MJS, Rain, AN, Kamel, AGM, Zahir, SIS, Khozirah, S, Hakim, SL, Zakiah, I, and
Azizol, AK. 2002. The screening of extracts from Goniothalamus
scortechinii, Aralidium pinnatifidum and Andrographis paniculata for anti-malarial
activity using the lactate dehydrogenase assay. J Ethnopharmacology 82: 239242.
Otake, T, Mori, H, Morimoto, M, Ueba, N, Sutardjo, S, Kusumoto, IT, Hattori, M, and
Namba, T. 1995. Screening of Indonesian plant extracts for Anti-Human
Immunodeficiency Virus-Type 1 (HIV-1) activity. Phytother Res 9: 6-10.
Panossian, A, Hovhannisyan, A, Mamikonyan, G, Abrahamian, H, Hambardzumyan, E,
Gabrielian, E, et al. 2000 Pharmacokinetic and oral bioavailability of
andrographolide from Andrographis paniculata (Nees) fixed combination Kan
Jang in rats and human. Phytomedicine 7: 351-64.
Pholphana, N, Rangkadilok, N, Thongnest, S, Ruchirawat, S, Ruchirawat, M, and
Satayavivad, J. 2004. Determination and variation of three active diterpenoids in
Andrographis paniculata. J Phytochemical Anal (In Press)
Puri, A, Axena, R, Saxena, RP, Saxena, KC, Srivastava, V, and Tandon, JS. 1993.
Immunostimulant agents from Andrographis paniculata. J Nat Prod 56: 995-999.
Rahman, NNNA, Furuta, T, Kojima, S, Takane, K, and Mohd, MA. 1999 Antimalarial
activity of extracts of Malaysian medicinal plants. J Ethnopharmacol 64: 249-54.
Rajani, M, Shrivastava, N, and Ravishankara, MN. 2000. A rapid method for isolation of
andrographolide from Andrographis paniculata (Kalmegh). Pharm Biol 38:
204-209.
- 20 -
Sahasitiwat, S. 2002. The study of acute cardiovascular toxicity of diterpenoid lactones
isolated from Andrographis panuculata (Burm.f.) Nees. Master Thesis, Mahidol
University, Bangkok, Thailand, ISBN 974-04-2008-7.
Saxena, S, Jain, DC, Gupta, MM, and Sharma, RP. 2000. High performance thin layer
chromatographic separation of hepatoprotective diterpenoids from Andrographis
paniculata. Phytochem Anal 11: 34-36.
Shen, YC, Chen, CF, and Chiou, WF. 2000. Suppression of rat neutrophil reactive
oxygen species production and adhesion by the diterpenoid lactone
andrographolide. Planta Med 66: 314-317.
Thamlikitkul, V, Dechatiwongse, T, Theerapong, S, Shantrakul, C, Boonroj, P, Punkrut,
W, et al. 1991 Efficacy of Andrographis paniculata Nees for pharyngotonsillitis in
adults. J Med Assoc Thai 74: 437-42.
Thanagkul, B and Chaichantipayut, C. 1985. Double-blind study of Andrographis
paniculata Nees and tetracycline in acute diarrhea and bacillary dysentery.
Ramathibodi Medical J 8: 57-61.
Tipakorn, N. 2002. Effects of Andrographis paniculata (Burm.F.) Nees on performance,
mortality and coccidiosis in broiler chickens. PhD Thesis. Institute of Animal
Physiology and Animal Nutrition, Georg-August-Universität, Göttingen, Germany.
Vedavathy, S and Rao, KN. 1991. Antipyretic activity of six indigenous medicinal plants
of Tirumala Hills, Andhra Pradesh, India. J Ethnopharmacology 33: 193-196.
Zhang, CY and Tan, BK. 1996 Hypotensive activity of aqueous extract of Andrographis
paniculata in rats. Clin Exp Pharmacol Physiol 23: 675-8.
Zhang, CY, Kuroyangi, M, and Tan, BKH. 1998. Cardiovascular activity of 14-deoxy11,12-didehydroandrographolide in the anaesthetized rat and isolated right atria.
Pharmacol Res 38: 413-417.
Zhang, CY and Tan, BKH. 1997. Mechanism of cardiovascular activity of Andrographis
paniculata in the anaesthetized rat. J Ethnopharmacology 56: 97-101.
Zhao, J, Yang, G, Liu, H, Wang, D, Song, X, and Chen, Y. 2002. Determination of
andrographolide, deoxyandrographolide and neoandrographolide in the Chinese
herb
Andrographis
paniculata
by
Micella
Electrokinetic
Capillary
Chromatography. Phytochem Anal 13: 222-227.
- 21 -
Development of Pharmaceutical Products from Medicinal Plants
Project Personnel
1. Project Consultants
1.1 Feasibility study for selected medicinal plants and field activity
(Faculty of Pharmacy, Mahidol University)
1. Assoc. Prof. Promjit Saralamp
(Dept. of Pharmaceutical Botany)
M.Sc. (Pharmacy)
2. Asst. Prof. Noppamas Soonthornchareonnon Ph.D. (Phytochemistry)
(Dept. of Pharmacognosy)
3. Asst. Prof. Sompop Prathanturarug
(Dept. of Pharmaceutical Botany)
Ph.D. (Pharmaceutical Biology)
1.2 Project Site Selection
1. Vice President for Special Activities, Chulabhorn Research Institute
(Mr. Damrong Ratanapanich)
2. CRI Staff
2.1 Coordinating Staff and Field Work
1.
2.
3.
4.
Ms. Krittika Polratana
Ms. Wandee Sirapat
Mrs. Sujunya Ruengverayut
Mr. Roengwit Phetkaochouy
M.Sc. (Toxicology)
M.Sc. (Toxicology)
M.Sc. (Agriculture)
B.Sc. (Agriculture)
2.2 Phytochemical group
1. Assoc. Prof. Vanida Bhavakul
2. Mr. Sanit Thongnest
3. Ms. Piyanun Boonprasert
Ph.D. (Organic Chemistry)
M.Sc. (Applied Chemistry)
M.Sc. (Petrochemistry)
2.3 Pharmacological group
1.
2.
3.
4.
5.
Dr. Nuchanart Rangkadilok
Ms. Nanthanit Pholphana
Ms. Sumitra Suntararuks
Ms. Luksamee Worasuttayangkurn
Ms. Sumontha Nookabkaew
6. Ms. Jittra Hun
- 22 -
Ph.D. (Agricultural and Food Systems)
M.Sc. (Microbiology)
M.Sc. (Toxicology)
M.Sc. (Toxicology)
M.Sc. (Applied Analytical and
Inorganic Chemistry)
B.Sc. (Fisheries)