Implementation of sustainable

development on Herbal

Medicines in Thailand related

to the new and trend issue at

the present

Kornkanok Ingkaninan

Faculty of Pharmaceutical Sciences , Naresuan University

biodiversity folklore wisdom

Natural Product Research in Thailand

Traditional Thai Medicines (TTM) is the integrated system influenced by Traditional Chinese Medicines (TCM) and Ayurvedic Medicine and also Buddhism philosophy, astrology and folklore belief.

Holistic approach.

Traditional Thai Medicine

The use of TTM was declined during the Westernized period. However, with the support of the government, TTM increases its popularity again.

The National Plan for 1992–1996 stated that “The promotion of people's health entails the efforts to develop traditional wisdom in health care, including Thai traditional medicine, herbal medicine, and traditional massage”

Vision: “To be the national authority on Thai traditional and alternative medicine for self-reliance on health care and promotion of sustainable economy”

-

Department of Thai Traditional and Alternative Medicines, Ministry of Public Health (2003)

Department of Thai Traditional and Alternative Medicines: Mission

To develop technical capacity in Thai traditional and alternative medicine by protecting, conserving and promoting Thai Traditional medicine knowledge, to promote and develop knowledge management system, and establish standards of Thai traditional and alternative medicine practices and services to be equivalent to those of modern medicine, for effective and safe use in the health system as alternative form of health care for people to choose from.

National List of Essential Medicines 71 herbal medicines have been registered.

List of medicinal plants for primary health care 57 medicinal plants were selected and recommended

for the treatment and relief of 19 groups of common minor symptoms and diseases.

Education system National licenses on Thai Traditional medicines

Bachelor/Master degree programs in Thai Traditional Medicines

Bachelor/Master degree programs in Applied Thai Traditional Medicines

The constitution of the Kingdom of Thailand (2017) stated that the country will support the utilization of Thai traditional medicines.

Thai herbs Innovations

Thailand 4.0 policy

Herbal cities

Standardization of herbal medicines

Drug discovery and developmentFrom laboratory to market

Nature is a great source of bioactive compounds

Synthesis/combinatorial librarynatural products

High throughput screening

Identification of structural leads

Lead optimization

analog synthesisbiological evaluation in vitro metabolism studyCADDS studies

Patent filing

chemical process developments

2nd, 3rd biological evaluations

formulation stability studies

metabolism andpharmacokinetic(MAP)

toxicology

Marketing

Clinical trial Phase II

Clinical trial Phase III

Clinical trial Phase IFirst time in human; safety, PK20-80 volunteers

Efficacy, side effect200-300 volunteers

Large efficacy, side effect 1000-3000 volunteers

10,000

250

5

1

compounds

1-3 years

3-6 years

2-3 years

less than 1 year

2-4 years

8-10+ years in total

Registration filing

Translational Research of

Natural Products

Traditional Medicines

Herbal Drugs

Herbal Food Supplements

Herbal Cosmetics

Drug (Modern Drugs)

Traditional Drugs

• Therapeutic usage can be claimed.

• Evidence of the traditional use for ≥15 years

• Active ingredients are the same as traditional formula.

• Therapeutic usage cannotbe claimed.

• Evidence of Efficacy /Safety

• Herbal extract/ semi-purified, purified compounds can be used.

• *ASEAN guideline

Herbal Food

Supplement

• Therapeutic usage can be claimed.

• Evidence of Efficacy /Safety (intensive clinical studies)

• Herbal extract/ semi-purified, purified compounds can be used.

• *ASEAN guideline

(Modern) Herbal Drugs

• Therapeutic usage must not be claimed.

• Evidence of Efficacy is not required.

• Herbal extract/ semi-purified, purified compounds can be used.

• *ASEAN guideline

Herbalcosmetics

Checklist

Requirement from FDA (registration)

Quality: raw material/finished product specification, stability test

Safety: preclinical studies/clinical studies/ bibliographic evidences

Efficacy: preclinical studies/clinical studies/ bibliographic evidences

What we (academic) want:

Novelty : drugs, models, methods, process

What the industry wants:

Meet market needs

Novelty

Industrial applicability

formulation

Herbal Food Supplement

-Stability

-Quality Control

-isolation and identification of bioactive compounds -quality control

-Plant identification-Good agricultural and cultivation practice (GACP)

Standardized extract/purified compounds

Toxicity evaluation

Preformulation study

Stability study

extraction

Bioactivity evaluation-in vitro/in vivo

Herbs

Optimization of extraction /isolation process

Clinical studies

Patent filing

Patent filing

Ethnopharmacology

formulation

Herbal Food Supplement

-Stability

-Quality Control

-isolation and identification of bioactive compounds -quality control

-Plant identification-Good agricultural and cultivation practice (GACP)

Standardized extract/purified compounds

Toxicity evaluation

Preformulation study

Stability study

extraction

Bioactivity evaluation-in vitro/in vivo

Herbs

Optimization of extraction /isolation process

Clinical studies

Ethnopharmacology

Quality

formulation

Herbal Food Supplement

-Stability

-Quality Control

-isolation and identification of bioactive compounds -quality control

-Plant identification-Good agricultural and cultivation practice (GACP)

Standardized extract/purified compounds

Toxicity evaluation

Preformulation study

Stability study

extraction

Bioactivity evaluation-in vitro/in vivo

Herbs

Optimization of extraction /isolation process

Clinical studies

Ethnopharmacology

efficacy

formulation

Herbal Food Supplement

-Stability

-Quality Control

-isolation and identification of bioactive compounds -quality control

-Plant identification-Good agricultural and cultivation practice (GACP)

Standardized extract/purified compounds

Toxicity evaluation

Preformulation study

Stability study

extraction

Bioactivity evaluation-in vitro/in vivo

Herbs

Optimization of extraction /isolation process

Clinical studies

Ethnopharmacology

safety

Case Study 1: Food supplement

Brahmi, a medicinal plant for

memory improvement

Herbal medicine for memory improvement

Brahmi (พรมม)ิ

Bacopa monnieri (L.) Wettst (Plantaginaceae)

• Ayurveda medicine for promoting mental health and

improving memory

• Memory enhancer (Kidd, 1997; Roodenrys et al, 2002)

• Antidepressant (Sairam et al, 2002)

• Increase verbal learning, memory consolidation, and

speed of early information processing in 40-65 years

old volunteers (Stough et al. 2001)

• Other activities such as anticonvulsive, vasodilating,

antioxidant and hepatoprotective activities

Traditional uses & Pharmacological Studies

Cultivation and harvesting

New indications

Preformulation

studies

RCT study

for effect on memory

(60 subjects)

Pharmacological studies:

effect on memory

Extraction and quality

control

Acute, sub-chronic &

chronic toxicity

Standardized

extract

Pharmacokinetics

Comparative study with Gingko

Metabolism/drug

interaction

Brahmi, a medicinal plant for memory improvement

Patent/

Technology transfer

1. Bacoside A3 - β-D-glc-(13)-O-[-L-ara-(12)]-O-β-D-glc

3. Bacopaside X --L-ara-(12)-[β-D-glc-(13)]--L-ara

2. Bacopaside II --L-ara-(12)-[β-D-glc-(13)]-β-D-glc

4. Bacopasaponin C -β-D-glc-(13)-[-L-ara-(12)]--L-ara

5. Bacopaside I --L-ara-(12)-[6-O-sulfonyl-β-D-glc-(13)--L-ara

Jujubogenin

Pseudojujubogenin

Structures of saponin glycosides from Brahmi

HPLC Chromatogram of brahmi extract (2 mg/ml)

Minutes

5.0 7.5 10.0 12.5 15.0 17.5

mA

U

0

50

100

150

Bacosid

e A

3B

acopa

sid

e II

UN

-4

Bacopa

sapo

nin

C

Bacopa

sid

e I

B

acop

asapo

nin

X

Standardized extract with total saponins not less than

5%

Petit Patent:Extraction process

Monograph of Brahmi

leaf

Whole plant flower root

flower fruit

Physicochemical parameters of Brahmi

Limit values in monographs

This study

(%w/w)

The BP

(%w/w)

The AP

(%w/w)Loss on drying Not > 11.0 Not > 11.0 Not reported

Total ash Not >13.0 Not > 13.0 Not > 18.0

Water-soluble extractive

Not < 15.0 Not < 15.0 Not < 15.0

Ethanol-soluble extractive

Not < 9.0 Not reported Not < 6.0

Total saponin Not < 1.5 Not < 1.0 Not reported

Good Agricultural and Collection Practice

(GACP)

Saponins in each part of Brahmi

0.00

1.00

2.00

3.00

shootlower part root

leaves

upper stemlower stem

Parts

sapo

nins

in B

rahm

i

(%w/

w dr

y wt

.)

Bacopaside I Bacopasaponin C Bacopasaponin C isomerBacopaside II Bacopaside A3

Phrompittayarat et al. Songklanakarin J Sci Technol. 2011. 32:193-199.

Stability studies

0

20

40

60

80

100

0 5 10 15 20 25 30Time (days)

% C

on

cen

tra

tio

n o

f B

aco

pa

sid

e I

5°C 40°C 60°C 80°C

0

20

40

60

80

100

0 5 10 15 20 25 30Time (days)

% C

on

cen

tra

tio

n o

f B

aco

sid

e A

3

5°C 40°C 60°C 80°C

Stability study of bacopaside I and bacoside A3

in crude brahmi extract at 5, 40, 60, and 80ºC

bacopaside I bacoside A3

O

OH

O

R

Jujubogenin

3

23

22

O

OH

O

R

Pseudojujubogenin

Phrompittayarat et al. Planta Med. 2008, 74:1756-63

Stability of bacopaside I in Brahmi extract at 40 º C

0

20

40

60

80

100

0 1 2 3 4 5 6

Time(Month)

%R

ela

tive o

f co

ncen

trati

on

Brahmi tablet formulation

Properties weight Disintegration Dissolution Hardness Thickness

Criteria 10 % 1.00 % < 20 min 6-7 Kg -----

Results 1.87 % 0.27% 10-12 min 6.45 Kg 5.107 mm

Evaluation Pass Pass Pass Pass Pass

containing 300 mg of standardized extract

in vitro/ in vivo/ in human

Morris Water Maze Test

Hidden Platform

Visual Cues

Escape latency

The effect of brahmi extract on rats expressed as escape latency

in Morris water maze test.

0

2

4

6

8

10

12

14

16

18

Single dose 1 week 2 weeks

Escap

e late

ncy (

seco

nd

s)

Control Vehicle Aricept BM 20 mg BM 40 mg BM 80mg

The effect of brahmi extract on cholinotoxin (AF) induced

amnesia rats expressed as escape latency in Morris water maze test.

0

10

20

30

40

50

60

Vehicle

Vehicle

+AF 6

4 A

Positive

+AF 6

4A

BM 2

0 mg+A

F 64A

BM 4

0 mg+A

F 64A

BM 8

0mg+A

F64A

Esc

ape

late

ncy

(sec

)

Uabundit et al. J Ethno Pharmacol. 2010, 127:26-31

0

5

10

15

20

25

30

35

40

CA1 CA2 CA3 DG

De

ns

ity

of

ch

oli

ne

rgic

ne

uro

ns

(ce

lls

/ 2

55

m

2 )

Control Vehicle Vehicle+AF 64A

Aricept+AF64A BM 40+AF 64A

# # #

#

# p-value .05 compared with vehicle

p-value .05 compared with vehicle+AF 64A

The effect of Bacopa extract on the cholinergic neurons

density in various subregions of hippocampus in Alzheimer’s

disease model induced by AF64A.

Effect of Brahmi extract on AChE activity

in Ab 25–35-treated cells

Primary cortical cells were treated with 50 M aggregated A25–35 in the absence and

presence of 100 g/ml Brahmi extract for 48 h. After treatments, cell viability and

intracellular AChE activity were determined from survival cells. The ratio of AChE

activity/cell viability was calculated and referred as AChE activity of individual cells.

Limpeanchop et al. J. Ethnopharmacol. (2008)120: 112-117

Brahmi vs Ginkgo and Donepesil 3 month oral administration of the extract/drug

demonstrated that 40 mg/kg Brahmi extract -treated group showed a similar efficacy of both spatial memory and object recognition as 60 mg/kg Ginkgo extract -treated and 1 mg/kg donepezil-treated groups which were significantly better than those of the vehicle control group.

Chronic daily dosing of Bacopa (40mg/kg),

Ginkgo biloba (60mg/kg) or donepezil (1mg/kg)

for 8 weeks. (A) Cerebral blood flow (B) Systolic BP

Brahmi increases

cerebral blood flow

in rat independent of

blood pressure.

.

Kamkaew et al. (2012) Phytother

Res.

Effect of Bacopa monnieri on the quality of life in the middle aged and elderly volunteers

Peth-Nui et al. Evid Based Complement Alternat Med. 2013;

Placebo Bacopa tablet

600 mg/day

working memory, ERP, Bond Lader, hematological changes & Clinical chemistry determination

60 volunteers(55-80 years old)

300 mg/day

Visit 3Visit 1

Visit 2

Visit 4Visit 5

Visit 6

1 week1 month

2 month3 month

1 month

-Bacopa tablet, Placebo

-Single Dose Effect

Baseline Data

Delay Effect

Determine Parameters Determine Parameters Determine Parameters

Results

-Taking Brahmi extract (300 and 600 mg/day) for 2 -3 months, physical fitness via the increase in efficiency of postural balance was improved.

- Taking Brahmi extract (300 and 600 mg/day) for 2 -3 months, psychological fitness was improved which can be observed in alertness, attention, learning and memory improvement.

-Brahmi extract (600 mg/day) was also able to decrease depression-like symptom.

-The dosage used in this study did not produce toxic and side effects.

in vitro/ in vivo/ in human

Toxicity study (OECD, 1981; WHO, 2000)

For Acute toxicity, no rats died in 2 weeks after 5

g/kg BW of Brahmi oral administration.

For Sub-chronic toxicity, no physical and pathological

changes when rats were fed with 4, 40 and 80 mg/kg

BW of Brahmi for 100 days.

For chronic toxicity, no behavior changes when rats

were fed with 30, 60, 300 and 1,500 mg/kg BW of

Brahmi for 270 days.

In the clinical trial, the dosage used in this study did not produce toxic and side effects.

Herb-Drug Interaction Effect of Brahmi extract on rat and human cytochrome P450

Brahmi extract showed low affinity on CYP1A2, CYP2C, CYP2E1 and CYP3A both in rat and human liver microsomes.

Significant drug interactions would not be expected.

Summary Quality:

GACP and monograph of Brahmi

Standardization of Brahmi extract (with markers).

Formulation of Brahmi tablet.

Efficacy:

The memory enhancing effect of Brahmi has been proven

in vitro, in vivo and in human.

Safety:

Brahmi extract showed no toxicity in rats/mice (OECD

guideline) and in human.

Bhrami has been used as vegetable and as drugs in Thai

traditional medicines and Ayurvedic medicines.

Technology Transfer

The preparation method of Bacopa extract was patented

in 2008.

We transferred the technology to the Governmental

Pharmaceutical Company (GPO) in Sep 2011.

Brahmi GPO was registered as a food supplement in

2013.

The herbal supplement of Brahmi was launched to the

market on Sep 3rd, 2014.

Brahmi-Fruit Essence

Source: Nugg Blog

Bacopaside A

Herbal Compress Anti-cellulite CreamSource: Nugg Blog

Dermato-Cosmetics

Efficacy and Safety Tests

Chemical-analytical Research

Source: Nugg Blog

Conclusion

• Herbal medicines are supported by Thai governments in terms of regulation and the public uses, education as well as R&D.

• Both governmental and private sectors in Thailand are interested in R&D of natural products.

• ASEAN guidelines are applied for the registration process.

Associate Professor P. Soralum, Mahidol University

B. Kiattanakorn, TCFF

D. Pongpaew, Herb for Health

Dr. S. Nimpitakpong, Phitsanuvej Hospital

Professor I. Khan, the National Center for

Natural Products Research, MS, USA

Faculty of Pharmaceutical Sciences,

Naresuan UniversityN. Limpeanchop

T. Teaktong

A. Sarapanchotiwittaya

S. Wittaya-areekul

J. Viyoch

S. Ooputthinan

D. Pekthong

W. Phromphitayarat

C. Tothium

S. Sookying

Faculty of Medical Sciences,

Naresuan UniversityP. Taepawarapruk

N. Taepawarapruk

S. Tanoi

S. Cheunchujit

S. Lerdtrakul

K. Chootip

Khonkaen UniversityW. Putalun

J. WattanathornS. Matchimpura

P. Rangseekajee

Medical Sciences Center,

PhitsanulokN. Nuengchamnong

Kyushu UniversityH. Tanaka

Université de Franche-ComtéC. Girard C. Demougeot

F. Muyard H. Martin

Vrije Universiteit, AmsterdamH. Irth J.De Vlieger

W.M.A. Niessen

Thammasat UniversityS. Sireeratawong

Acknowledgement

Thank you for your attention