research and development on herbal product for aging society · impairment using force plate...

Research and Development on

Herbal Product for Aging Society

Krittiya Thisayakorn, Ph.D.

Senior researcher

Expert Center of Innovative Herbal Product

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InnoHerb Introduction

Aging society

Braini-Tab for memory

• Rationale

• Research questions

• Clinical study

• Conclusion

ParkinPas for Parkinson

• Rationale

• Research questions

• Clinical study

• Conclusion

Outline

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Expert Center of Innovative Herbal Products

(InnoHerb)

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MISSION

Conducting R&D projects of health products from Thai

medicinal plants i.e. phytomedicines, dietary supplements and

cosmeceutics.

Providing consultancy expertise in R&D contract research,

training and technology transferring.

Analysis and testing services ie. analysis of active

ingredients, anti-microbial activity testing, and

pharmacological & toxicological testing of extract and

finished products.

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Lab

Extraction

Analysis of

Active compounds

Efficacy study

Market

Technology Transfer

Stability study

FormulationSafety Evaluation

ACTIVITIES

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FACILITIES

Phytochemistry lab

Pharmacology lab

Microbiology Lab

Toxicology lab- In vivo - In vitro ( genotoxicity lab)

Formulation Lab

Analytical lab

(LC-MS,GC-MS,HPLC, GC,)

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CISSA-ROIDS

(Hemorrhoid)

NUTRI-GAC (Antioxidant)Brainni-Tabs

(Memory Improvement)

Dietary Supplement products

“VITISTRA” Thai Grape Seeds

4GRAINE(Migraine)

Power Plus SiRelax induce relaxation and sleep

Heath products from essential oil

Anti-pyoderma gel

and oil for dogs

Herbal Stream Sauna Powder

(Relaxation)

Aromatherapy Liquid Soap

Herbal Compress gel

999

Cosmeceutics

LICO-SCAR CREAM

HAIR TONIC (ANTI-DANDRUFF)

ANTI-MELASMA /UV-PROTECTION

Phyllanthus emblica Linn

CAJAN PEPTIDE SERUM

ANTI-ACNE

SKIN ANTIOXIDANT

Anti-sprain BP Adaptogenic

Anti-ulcerogenic Anti-stress Weight control

Greater Pharma Ltd, Co.

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Aging Society

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https://www.adb.org/features/asia-s-growing-elderly-population-adb-s-take

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พ.ศ. 2556(2013) พ.ศ. 2558(2015) พ.ศ. 2563(2020) พ.ศ. 2568(2025) พ.ศ. 2573(2030)

สังคมสูงวัย (aged society)

สังคมเยาว์วัย (young society) สังคมสูงวัยอย่างสมบูรณ์ (complete aged society)

สังคมสูงวัยระดับสุดยอด (super aged society)

ที่มา : 1) พ.ศ. 2556 ข้อมูลประชากรผู้ที่มีสัญชาติไทยและมีช่ืออยู่ในทะเบยีนบ้านจากทะเบียนราษฎร์ ของสานักบริหารการทะเบียน กรมการปกครอง

2) พ.ศ. 2558 - 2573 ข้อมูลการคาดประมาณประชากรของประเทศไทย พ.ศ. 2553 - 2583 จาก สานักงานคณะกรรมการพัฒนาเศรษฐกิจและ

สังคมแห่งชาติ

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https://sdthailand.com/tag/good-living-for-aging-society/

Good living…Aging Society

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AC

12

3+

-

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Aging society

Common neurodegenerative disease

Alzheimer’s disease; the 1st common found

Incidence 50-60%

Affects memory

Rationale

https://www.123rf.com/photo_61625061_stock-vector-lost-grandmother.html

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Why is spinach interesting?

- Rich source of vitamins;

B9 or folate

- Folate or folic acid is an

essential vitamin

- Necessary for the brain

function

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1. Can spinach improve cognitive impairment

in animal model?

2. What is a possible mechanism of spinach?

3. Does spinach show any signs of toxicity?

Research questions

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Operant conditioning system

Reward

Stimulus light

Hopper

Lever

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FR condition

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VR condition

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WR

0 1 2 3 4

Cycles

0

2

4

6

8

Nu

mb

er

of

lev

er

pre

sses

WRSP12

WRSP6

WRFO

WRDW

0 1 2 3 4

Cycles

0

10

20

30

40

Nu

mb

er

of

lev

er

pre

sses

VRSP12

VRSP6

VRFO

VRDW

VR

*

*

FR

0 1 2 3 4

Cycles

0

5

10

15

20

25

Nu

mb

er

of

lev

er

pre

sses

FRSP12

FRSP6

FRFO

FRDW

*

DW

0.5 mg/kg Folic acid

125 mg/kg Spinach

250 mg/kg Spinach

* P<0.05

*

1. Can spinach improve cognitive impairment in

animal model? Yes.

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2. What is a possible mechanism of spinach?

Choline

Acetyl CoA

Acetylcholine

Acetylcholine in vesicle

Acetylcholine receptor

Acetylcholine binds to receptor

Acetylcholinestease

Braini-Tab

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2. What is a possible mechanism of spinach?

Possibly due to acetylcholinesterase inhibiting effect

***

0 1 2 3 4 5

group

0

500

1000

1500A

Ch

E l

ev

els

(U

/L)

* P < 0.05

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Choline

Acetyl CoA

Acetylcholine

Acetylcholine in vesicle

Acetylcholine receptor

Acetylcholine binds to receptor

Acetylcholinestease

Braini-Tab

A possible mechanism

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Acute oral toxicity : LD50>2,000 mg/kg

Cytotoxicity test

L929: IC50 = 33.21mg/ml

HepG2: IC50 > 1000 µg/ml

3. Does spinach show any signs of toxicity?

No

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Formulation

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Clinical trials

Volunteers: Division of Neurology, Phramongkutklao Hospital

Inclusion criteria: age 55 years, willing to participate, mild memory,

informed consentExclusion criteria: severe dementia, severe depression, severe medical

conditions

Subject: 3 groups (40/group), high dose (9 tab/d), low dose (6 tab/d),

placebo

Memory function: Montreal cognitive assessment (MoCA) test

Psychomotor speed: Grooved pegboard (GP), Trail making,

Digit symbol tests

The study period: 3 visits

The follow up period: 1-month (Visit 2) and at 3-month (Visit 3)

Laboratory tests: CBC, BS, BUN, Cr, uric acid, AST, ALT,

electrolytes

A double-blind randomized placebo-controlled study

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Visit 1

• Demographic

• Stratified randomization

• Explained how to take medication

• Blood test

• MoCA test

• Groove pegboard test

• Trail makings test

• Digit symbols test

Visit 2

(1-month)

• Subjective change

• Compliance

• Adverse event

• MoCA test

• Groove pegboard test

• Trail makings test

• Digit symbols test

Visit 3

(3-month)

• Subjective change

• Compliance

• Adverse event

• Blood test

• MoCA test

• Groove pegboard test

• Trail makings test

• Digit symbols test

** *

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Worse

Better

Better

WorseMoCA Test

GP Test (sec.)

***

*

*p<0.05

**

*

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Trail making test (sec.)

Digit symbol test

Worse

Better

Better

Worse

*p<0.05

* **

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Parameter LS HS PL

Visit1 Visit3 Visit1 Visit3 Visit1 Visit3

Hematocrit 384 374 385 384 383 383

WBC 7,200 6,900 6,650 5,800 5,900 6,450

Platelet 2.47x105 2.46x105 2.34x105 2.42x105 2.48x105 2.46x105

Blood sugar 10532 10421 9717 10319 11040 10519

BUN 172 144 155 144 155 203

Creatinine 0.90.3 0.90.4 0.90.3 0.90.2 0.90.3 0.90.3

Uric acid 5.51 6.12 5.82 5.91 5.92 6.11

AST 245 256 246 247 236 237

ALT 218 239 2410 2411 219 2211

Na 1402 1402 1403 1402 1392 1412

K 40.4 40.5 40.5 40.6 40.4 40.4

Cl 1033 1033 1033 1032 1033 1033

CO2292 283 283 275 282 273

Safety analysis

No abnormal laboratory finding

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Spinach supplements in aged rats possibly aid

the promotion of brain functions, especially

learning and memory via operant condition

system, involved cholinergic pathway.

Spinach supplement for 3 months was safe to

use and possibly improve cognitive function in

volunteers.

Conclusion

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Braini-Tab

TISTR

2552-2555

MDmate

Anajak Intertrade Co., Ltd.

2557

Transferred technology

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Rationale

A common degenerative neurological disorder

Affects motor functions

The incidence of PD is approximately 1.5 million

Symptoms related to dopamine depletion

(Bradykinesia, rigidity, tremor or postural

instability)

https://drmwhite.com/parkinsons-disease/

Drug of choices are dopaminergic

agents

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Passiflora foetida (passionflower) contains

ingredients of alkaloids (harmala alkaloids)

Why is passionflower interesting?

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1. Can passionflower improve motor function

impairment using force plate actimeter technique?

2. What is a possible mechanism of passionflower?

3. Does passionflower show any signs of toxicity?

Research questions

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Interface

Force plate

Transducer

Force plate actimeter

Data analysis

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Powe

r Spe

ctra

Frequencies (Hz.)

MPTP onlyMPTP+Sinemet

a = P< 0.05 compared with control

* = P< 0.05 compared with MTPT only

1. Can passionflower improve motor function

impairment using force plate actimeter technique?

Yes

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a = P< 0.05 compared with sham

b = P< 0.05 compared with control

Dopamine Monoamine oxidase

2. What is a possible mechanism of passionflower?

More dopamine in

the synaptic cleft

A possible mechanisms of action

Presynaptic neurons

Postsynaptic neurons

vesicle

DA receptor

DA

MAO

ParkinPas

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Acute oral toxicity : LD50>6,000 mg/kg

Subchronic toxicity : did not cause any

significant disorders in rats in terms of body

weight, food consuming rate, hematology, the

clinical chemistry in blood and urine, and the

pathology of major organs.

3. Does passionflower show any signs of toxicity?

No

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Formulation

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Clinical trials

Randomized double-blind placebo controlled study

Volunteers: Division of Neurology, Phramongkutklao Hospital

Inclusion criteria: age 30 years, idiopathic Parkinson’s disease,

Hoehn & Yahr (H&Y) stage 0.5-2.5, maintain

prescribed, informed consent

Exclusion criteria: severe dementia, severe medical, psychological or

oncological conditions

Subject: 3 groups; ParkinPas (high dose 9 tab/d+ low dose 6 tab/d),

placebo

Motor function: Clinical, Unified Parkinson’s disease rating scale

(UPDRS) score

The study period: 3 visits

The follow up period: 1-month (Visit 2) and at 3-month (Visit 3)

Laboratory tests: CBC, Cr, uric acid, K, AST, ALT,

lipid profiles, HbA1C

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ParametersPlacebo

(n 29)

ParkinPas

(n 45)

p-valueb

(between

group)

p-valueb

(between

group)

Mentation- behavior-

mood

visit 1 1(0-3), 0.93 1(0-7), 1.44 0.617

visit 2 0(0-3), 0.58 0.5(0-6), 0.92 0.815 0.769

visit 3 0(0-2), 0.25 0(0-4), 0.54 0.689 0.867

p-valuea(within group) 0.513 0.007*

ADL

visit 1 8(2-21), 9.55 8(1-22), 9.16 0.574

visit 2 2(0-6), 2.25 2(0-30), 2.89 0.841 0.205

visit 3 5(1-21), 7.56 7(1-20), 7.37 0.915 0.714

p-valuea(within group) < 0.001* < 0.001*

Motor

visit 1 14(3-39), 15.79 13(2-43), 15.36 0.716

visit 2 15(3-39), 16.45 12(1-26), 13.00 0.207 0.627

visit 3 12.5(2-33), 14.06 13(3-22), 13.11 0.855 0.114

p-valuea(within group) 0.442 0.308

Complication

visit 1 0(0-2), 0.62 1(0-7), 1.13 0.949

visit 2 0(0-4), 0.50 0(0-30), 1.37 0.506 0.404

visit 3 0(0-3), 0.50 1(0-11), 1.29 0.218 0.572

p-valuea(within group) 0.186 0.15

Total

visit 1 25(7-63), 26.9 24(3-72), 27.10 0.976

visit 2 18.5(3-48), 19.80 16(1-70), 18.18 0.411 0.761

visit 3 20(3-54), 22.38 23(5-41), 22.31 0.750 0.440

p-valuea(within group) 0.078 < 0.001* 46

10

15

20

25

30

visit 1 visit 2 visit 3

Total

Placebo ParkinPas

0

2

4

6

8

10

12

visit 1 visit 2 visit 3

Activity of Daily Living

Placebo ParkinPas

0

0.5

1

1.5

2

visit 1 visit 2 visit 3

Mentation- behavior-mood

Placebo ParkinPas

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Parameter Low dose

Visit 1

Low dose

Visit 3

High dose

Visit 1

High dose

Visit 3

Placebo

Visit 1

Placebo

Visit 3

Hematocrit (%) 38.2 + 2.0 38.1 + 1.1 39.2 + 2.2 40.0 + 3.0 41.1 + 2.3 40.3 + 2.5

WBC (/cumm) 7,800+20.1 8,200 + 15 6,800+22.0 7,200 +10 9,200+7,9 8,900+ 10

Platelet (/cumm) 200,400 + 200 250,000 +

323

330,000 +

421

320,123 +

211

245,000 +

321

321,000 +

321

Creatinine 1.2 + 1.0 1.1 + 0.8 0.8 + 0.5 0.9 + 1.0 0.8 + 1.0 1.1 + 0.9

Uric acid 5.5 + 1.3 5.2 + 0.1 5.1 + 0.8 5.1 + 1.1 6.0 + 1.0 6.7 + 0.8

Potassium 3.7 + 1.4 3.8 + 0.1 4.2 + 1.5 4.6 + 1.2 3.8 + 1.7 4.2 + 1.8

Triglyceride 135 + 20.3 140 + 21.0 150 + 23 160+20.3 140 + 30 148 + 16

HDL 52.3 + 12 47 + 21 55.4 + 10 58.2 + 8.0 46.8+10.1 50.1 + 12

LDL 140 + 12 146 + 13 158 + 21 162 + 21 155 + 24 160 + 31

AST 23.6 + 1.0 32.1 + 1.8 34.2 + 3.2 39.1 + 36 41.2 + 4.1 30.1 + 2.8

ALT 29.9 + 2.1 34.9 + 3.9 41.3 + 1.8 40.2 + 8.9 43 + 3.6 37.8 + 5.7

HbA1C 5.4 + 2.1 5.8 + 2.9 5.2 + 1.4 5.7 + 3.6 5.1 + 0.8 5.3 + 2.2

Safety analysis

No abnormal laboratory finding

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Conclusion

Passionflower effect on relieving Parkinson’s

symptoms for better performance in mice

possibly relate dopaminergic pathway

ParkinPas supplement for 3 months was safe

to use and appeared more effective than

placebo as an adjunctive treatment in patients

with Parkinson’s disease

49https://sdthailand.com/tag/good-living-for-aging-society/

Good living…Healthy aging

Summary

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Neurologist staffs

Division of Neurology, Phramongkutklao Hospital

Researchers and co-workers

Expert Center of Innovative Herbal Product, TISTR

Acknowledgement