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以功能性生物標記平台開發
老化生技產品
蘇益仁 教授
國家衛生研究院 特聘研究員
成功大學 講座教授
南臺科技大學 講座教授
21世紀疾病與失智症有共通的起源-飲食失調
Major Diseases in 21st Century: Metabolic
syndrome (obesity), Cancers, and Ageing
Infection
ER Stress
(內質網壓力)
Genetics
Stress
Nutrition
Inflammation (發炎)
Metabolic Syndrome
(代謝症候群)
Oxidative
Stress
(氧化壓力)
Genomic
instability
DNA damage
Ageing
(氧化DNA傷害)
(基因體不穩定)
Repair (修護DNA)
WHO Recommendation:
Diet & Exercise
Cancer 癌
Alzeimer’s
disease
from Non-Alcoholic Fatty Liver to Cancer Development
(1) Non-alcoholic
fatty liver to HCC;
(2) HCV HCC
(3) HBV HCC
Cancer
Science 332(24), p1519, 2011
Revolution Concept on the Role of Foods in Health
食物在健康所扮演的革命性新觀念
Revolution Concept on the Role of Foods in Health
食物在健康所扮演的革命性新觀念
Health Food is classically considered to be:
Anti-oxidants, vitamins, trace metals,
fiber, essential fatty acids--etc.
? Are there functional factors which play a role like
drugs or have proven therapeutic effects ?
How can we step forward ?
Natural Products and Healthy Ageing
天然食材與養生長壽
PPAR ligands:
L: ligands
B: binding
proteins
Co-act: Co-activator
白藜蘆醇
(Resveratrol)
薑黃
(Curcumin)
不飽和脂肪酸 (Omega 3)
類胡蘿蔔素(Carotenoids)
類黃酮(Flavonoids)
三萜類(Tripternoids)
B
PGC-1A
L
B
L
L
L
PPARs
RXRa
Bexarotene
Retinoids
Co-Act
PPRE
 PPAR a/g ↑
 NFkB ↓
 Apo E ↑
 mTOR ↓
代謝
抗發炎
抗癌
抗發炎
抗癌
抗老化
防癌、抗癌
8
The key regulator of metabolic
syndrome, inflammation, and cancers:
PPARs ( peroxisome proliferator activated
receptors ): 三種異構物 3 isoforms ( alpha, beta,
gamma)
Effects of PPAR agonists: lipid regulation,
Anti-inflammation, and anti-cell proliferation
Drugs with PPAR agonistic activities
PPARγ活化劑已開發成為糖尿病用藥
Takeda pioglitazone
Glaxosmithkline
Rosiglitazone
PPARs platform technology to develop
functional product from herbs
PPARs promoter plasmids
Extracts from
natural products
HepaG2 cell
Extracts of Natural products
Screening for products with
high PPAR agonistic
activities
從數百種天然食材中檢測
PPARs的活性大小
PPARγ activation fold compared to troglitazone(0.25ug/ml)
Combination of the L,B,C,S showed the best synergistic
activation of PPARα and PPARγ
Efficacy tests of Metamin 3D—animal and human
trials
Metamin 3D Clinical Trails in VGH,
Taichung, for metabolic syndrome
委託台中榮總進行雙盲試驗─
人體健康3D改善代謝症候群

Randomized, placebo-controlled in 2 groups

採盲式隨機分成兩組,試驗組給予健康3D,對照組給予安慰
劑。

Oral, tid, for 3 months


一天一次,一次三錠
給予試驗產品12週(3個月)
Publication and Patent
under application
Natural products for anti-oxidant, ageing,
anti-inflammation, and chemoprevention
Resveratrol: Red Grape Magic
• Red wine 5mg/bottle
Resveratraol
Science 1997: Resveratrol is effective for tumor
control at the stages of tumor
initiation, promotion, and
prevention
Synergistic effect of PPAR agonists
( omega 3 ) and resveratrol in
PPARγ activation
Omega 3
resveratrol
PPARγ activation by Metamin 3D and resveratrol
12
10
luciferae fold
8
6
4
2
0
Co
ntr
o
3D
l
20
ul
3D
20
ul
3D
+R
es
5u
l
20
ul
3D
+R
es
10
ul
20
ul
+R
e
s2
0u
l
Fig 2. PPAR gamma activation fold of six selected natural products
.
pioglitazone 0.1uM were used as positive control. Luminescence Values are means ±
SEM, n = 3, and are folds of the control (DMSO) mean.
resveratrol
2.00
AU
1.50
resveratrol
1.00
0.50
1.50
Standard resveratrol 0.5 mg/ml
resveratrol
0.40
AU
0.30
0.20
AU
0.00
0.00 10.00 20.00 30.00 40.00 50.00 60.00
Minutes
1.00
0.50
0.00
0.00 10.00 20.00 30.00 40.00 50.00 60.00
Minutes
綠加 3D+R plus HPLC 圖譜
0.10
0.00
0.00 10.00 20.00 30.00 40.00 50.00 60.00
Minutes
Standard resveratrol 0.1 mg/ml
Resveratrol 含量經比對標準品後,
為8.421896 mg / 1g , 大於規格的
6mg/ g
Curcumin has antioxidant, anti-inflammatory, and
anti-amyloid activity and is a promising compound
for the development of AD therapies
Curcuma longa (薑黃)
 In vitro studies with curcumin
-- Antioxidant effect
-- Anti-inflammatory effect
-- Inhibition of b-secretase
-- Inhibition of Ab aggregation
 In vivo studies with curcumin
-- Inhibition of Ab deposition
-- Inhibition of Ab oligomerization
(薑黃素)
(CNS Neuroscience&Therapeutics 16, 285-297, 2010)
Question
The mechanism by which
curcumin inhibits Ab deposition
remains to be clarified
Curcumin Clinical Trials
Cancer
• Colorectal cancer
• Pancreatic cancer
• Breast cancer
• Prostate cancer
• Multiple myeloma
• Lung cancer
• Cancer lesions
• Head and neck cancer
•Vitiligo
•Psoriasis
Inflammatory diseases
•Crohn disease
•Ulcerative proctitis
• Ulcerative colitis
•Inflammatory bowel disease
• Irritable bowel
syndrome
•Rheumatoid arthritis
• Osteoarthritis
• Chronic anterior uveitis
•Recurrent anterior uveitis
•Post operative Inflammation
• Gastric ulcer
• Peptic ulcer
•H. pylori infection
•Idiopathic orbital inflammatory
Pseudotumor
Skin diseases
Neurodegenerative diseases
•Dejerine-Sottas disease
• Alzheimer's disease
O
O
Cardiovascular diseases
OCH3
H3CO
HO
OH
Curcumin
•Acute coronary syndrome
• Atherosclerosis
Metabolic diseases
• Diabetes
•Diabetic nephropathy
•Diabetic microangiopathy
• Lupus nephritis
Renal diseases
•Renal transplantation
Viral diseases
OTHERS
b-Thalassemia
•Biliary dyskinesia
•Gallbladder
contraction
•Recurrent
respiratory tract
infections
• Cholecystitis
•Hepatoprotection
•Chronic arsenic
exposure
• Alcohol intoxication
•Chronic bacterial
prostatitis
•Acquired immunodeficiency
syndrome
Gupta, Patchva and Aggarwal:, AAPS J. (in
press)
阿茲海默症的學理基礎:
異常構造的β類澱粉蛋白(β -amyloid)無法清除,而累積在神
經細胞中形成細纖維團(fibrillary tangle)或細胞外的斑塊(plague)
,引起神經細胞傷害或死亡,導致傳導功能、發炎等病變。
Two Key Issues for Curcumin in
Drug Development
No drug has been ever developed
1. No patent:
a. Clarify the signal pathway and establish a
reporter assay to screen the drug candidates
b. Modification of curcumin as a new drug
2. Poor absorption and difficult to reach
PK value for drug use: use new delivery
system such as lipoid carrier, or to modify
curcumin to be water soluble
Curcumin showed PPARg activity
in HepG2 cells
** Curcumin-Indena
The effects of natural compounds on the anti-oxidant activity
by assay of NrF2 in Huh7 cells.
Method: Luciferase Reporter Assay
Huh7-24hr
Relative luciferase activity
NrF2 activity
5
4
3
2
1
0
Control
Curcumin 15μ g/ml(Indena)
Resveratrol 25μ M
Preliminary data
Curcumin could significantly increase
ApoE protein expression in HuH-7 cells
HuH-7 24h
Western blot assay
RXRα
ApoE
β-actin
Rapamycin
PPARα
DMSO
Untreated
p-mTOR
Sigma
Curcumin
Indena
Curcumin
5
5
10
20
10
20
μg/mL
Apo E and Alzheimer's disease (AD)
Apolipoprotein E (Apo E) is important in plasma lipid
metabolism. Apo E polymorphism or genotype is
closely related to total plasma cholesterol levels and
LDL levels: E2 allele is associated with lower , and
E4 with higher levels, compared with common E3
allele.
(Hagberg J.et al : Physiol. Genomics 2000;4:101-108)
The risk of developing AD: Apo E4 ˃ E3 ˃ E2 (84yrs,
75yrs,68yrs)
Mariani SM, Medscape General Medicine 2004;6:47
Cramer PE., et. al. Science 2012, 335: 1503-1506.
The flux of Aβ/cholesterol/ApoE through the cell
and the potential regulation by PPAR/RXR/LXR
PPAR
(Wolozin B, Neuron 41, 7-10, 2004)
膽固醇(cholesterol)代謝與
AD的致病機制息息相關
1. Total cholesterol及LDL在AD病人顯著上升
,且與Aβ42的形成相關(Kuo,1998).
2. 降低Cholesterol的藥物Statins可降低A β及
App片斷(β -CTFs).
3. ApoE可運送cholesterol及結合LDL受體,與
Aβ的沉積相關(Striffmater,1993).
4. Ceramide及cholesterol代謝異常可引發
oxidative stress並引起大腦aging及AD.
Hypothesis
Curcumin may induce Ab clearance through
RXR/LXR-mediated apoE upregulation,
ameliorating AD pathophysiology
Curcumin
ApoE
Ab
clearance
Key Biomarkers for Health: PPAR/RXR and LXR/RXR
Regulated by PPAR agonist, Resveratrol and Curcumin
L
Oxysterol,
Curcumin
LXR
L
Glucose regulation
Lipid metabolism
Anti-cancer
Anti-inflammation
Alzeimer’s disease
L
RXRa LXR
Glucose regulation
Cholesterol metabolism
ApoE
The effects of Green Cross natural compounds on the
activity of PPARγ in Huh7 cells.
Method: Luciferase Reporter Assay
Huh7-24hr
Relative luciferase activity
PPARg activity
9
8
7
6
5
4
3
2
1
0
3D, 3DR, new product 均為一天之劑量
Control
50%EtOH
Troglitazone 0.125g/mL
3D
3DR
New product
3DRC 1顆
3DRC 2顆
3DRC 3顆
3DRC 4顆
The effects of natural compounds on the activity of PPARγ in Huh7 cells:
A comparison between 3DRC and Lipoguard ( Viva )
Method: Luciferase Reporter Assay
Huh7-24hr
Relative luciferase activity
PPARg activity
8
7
6
5
4
3
2
1
0
Control
50%EtOH
Troglitazone 0.125μ g/mL
3D
3DRC 1X
3DRC 4X
1040210
LipoGuard(上 )
LipoGuard(下 )
The effects of natural compounds on the activity of PPARγ in Huh7 cells.
Method: Luciferase Reporter Assay
Huh7-24hr
Relative luciferase activity
PPARg activity
8
7
6
5
4
3
2
1
0
Control
50%EtOH
Troglitazone 0.125μ g/mL
3D
3DRC 1X
3DRC 4X
1040210
LipoGuard(上 )
LipoGuard(下 )
The effects of Green Cross natural compounds on the
activity of ApoE in Huh7 cells.
Method: Luciferase Reporter Assay
Huh7-16hr
Relative luciferase activity
ApoE activity
7
6
5
4
3
2
1
0
3D, 3DR, new product 均為一天之劑量
Control
50%EtOH
Curcumin 15g/mL
3D
3DR
New product
3DRC 1顆
3DRC 2顆
3DRC 3顆
3DRC 4顆
服用3D+R plus Curcumin
就如每天喝一瓶紅酒、三餐吃咖哩、
喝綠茶、豆漿、綠藻精及芝麻
融合 地中海、日本、印度等國家長壽
的祕密。
「君、臣、佐、使」複方概念是二十一世紀生技
產品開發的主流。
Nature 2011
One for all and all in one