research and development on herbal product for aging society · impairment using force plate...
TRANSCRIPT
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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
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Heath products from essential oil
Anti-pyoderma gel
and oil for dogs
Herbal Stream Sauna Powder
(Relaxation)
Aromatherapy Liquid Soap
Herbal Compress gel
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Cosmeceutics
LICO-SCAR CREAM
HAIR TONIC (ANTI-DANDRUFF)
ANTI-MELASMA /UV-PROTECTION
Phyllanthus emblica Linn
CAJAN PEPTIDE SERUM
ANTI-ACNE
SKIN ANTIOXIDANT
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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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** *
30
Worse
Better
Better
WorseMoCA Test
GP Test (sec.)
***
*
*p<0.05
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**
*
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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?
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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
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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
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