-25.0

-20.0

-15.0

-10.0

-5.0

0.0

5.0

0w 4w 8w 12w

全脂肪面積変化量

対照飲料

被験飲料

***,###

***,###

(cm2)

-25.0

-20.0

-15.0

-10.0

-5.0

0.0

5.0

0w 4w 8w 12w

内臓脂肪面積変化量

対照飲料

被験飲料

***,##

***,###

(cm2)

** *****

-25.0

-20.0

-15.0

-10.0

-5.0

0.0

5.0

0w 4w 8w 12w

皮下脂肪面積変化量

対照飲料

被験飲料

#

**,###

(cm2) *

●●

●

●

●

●

●

●

●

●

●

●

###：P<0.001, ##:P<0.01, #:P<0.05 vs. placebo、***：P<0.001, **：P<0.01, *：P<0.05 vs. 0w

-25.0

-20.0

-15.0

-10.0

-5.0

0.0

5.0

0w 4w 8w 12w

全脂肪面積変化量

対照飲料

被験飲料

***,###

***,###

(cm2)

-25.0

-20.0

-15.0

-10.0

-5.0

0.0

5.0

0w 4w 8w 12w

内臓脂肪面積変化量

対照飲料

被験飲料

***,##

***,###

(cm2)

** *****

-25.0

-20.0

-15.0

-10.0

-5.0

0.0

5.0

0w 4w 8w 12w

皮下脂肪面積変化量

対照飲料

被験飲料

#

**,###

(cm2) *

●●

●

●

●

●

●

●

●

●

●

●

###：P<0.001, ##:P<0.01, #:P<0.05 vs. placebo、***：P<0.001, **：P<0.01, *：P<0.05 vs. 0w

Obesity is one of the most important worldwide health problems. Previous studies have reported that consumption of flavonoids in the diet is inversely correlated

to lower morbidity and mortality from coronary heart disease and many studies have shown that quercetin might possess beneficial effects on lipid and glucose

metabolisms. These studies suggest that quercetin is promising as new pharmacologic agent for the treatment of obesity-related health problems such as

metabolic syndrome. Thus, we investigated the impact of quercetin glucosides(QG), which are enzymatically trans-glycosylated isoquercitrin from natural sources

and have improved bioavailability, on visceral fat mass in diet-induced obese mice and the obese human.

Introduction & Objection

This study suggested that the quercetin glucosides activate lipolytic pathway in adipose tissues and are beneficial to improve obesity in human.

P362

Conclusion

Quercetin glucosides reduced visceral fat mass in healthy obese human Norifumi Tateishi1, Kahori Egawa1, Makiko Yoshimura1, Noriyuki Kanzaki1, ○Yoshinori Kitagawa1, Yoshinobu Kiso1, Soichiro Enomoto2, Masataka Sata3

1 SUNTORY WELLNESS Limited, Institute for Health Care Science, Osaka, Japan2 Department of Cardiovascular Medicine, University of Tokyo Graduate School of Medicine, Tokyo, Japan3 Department of Cardiovascular Medicine, Institute of Health Biosciences, The University of Tokushima Graduate School

Profile of Quercetin

Material

Preparation of Quercetin Glucosides(QG)

Exp.1：Animal study (The Effects of QGs on the diet-induced obese mice)

Protocol

ND HFHF+

0.05% QGHF+

0.25% QGHF+

1.25% QG

Casein 14.00 23.31 23.30 23.25 23.01L-Cystine 0.18 0.35 0.35 0.35 0.35Corn Starch 62.07 8.48 8.48 8.46 8.38Maltodextrin - 11.65 11.65 11.62 11.51Sucrose 10.00 20.14 20.13 20.09 19.88Cellulose 5.00 5.83 5.82 5.81 5.75Soy bean oil 4.00 2.91 2.91 2.91 2.88Lard - 20.68 20.67 20.63 20.42Mineral Mix 3.50 5.24 5.24 5.23 5.18Vitamin Mix 1.25 1.40 1.40 1.39 1.38Quercetin glucosides（QG） - - 0.05 0.25 1.25

Energy (kcal/g diets) 3.62 4.73 4.73 4.72 4.67

※Diets used in these experiments were D12451based pellet diets

(45kcal fat %, rodent diet formulation from RESEARCH DIETS inc.）

ND HFHF+

0.05% QGHF+

0.25% QGHF+

1.25% QG

Casein 14.00 23.31 23.30 23.25 23.01L-Cystine 0.18 0.35 0.35 0.35 0.35Corn Starch 62.07 8.48 8.48 8.46 8.38Maltodextrin - 11.65 11.65 11.62 11.51Sucrose 10.00 20.14 20.13 20.09 19.88Cellulose 5.00 5.83 5.82 5.81 5.75Soy bean oil 4.00 2.91 2.91 2.91 2.88Lard - 20.68 20.67 20.63 20.42Mineral Mix 3.50 5.24 5.24 5.23 5.18Vitamin Mix 1.25 1.40 1.40 1.39 1.38Quercetin glucosides（QG） - - 0.05 0.25 1.25

Energy (kcal/g diets) 3.62 4.73 4.73 4.72 4.67

※Diets used in these experiments were D12451based pellet diets

(45kcal fat %, rodent diet formulation from RESEARCH DIETS inc.）

Diet formulation

##:p<0.05, ＃；p<0.01 vs. HF (One-way ANOVA)

HF＋0.05%QG

HF

ND

HF＋0.25%QG

HF＋1.25%QG

20

25

30

35

40

45

0 10 20 30 40 50 60 70 80 90

Day

Body

weig

ht

(g)

#

#

####

#

####

##

##

##

#####

#

##:p<0.05, ＃；p<0.01 vs. HF (One-way ANOVA)

HF＋0.05%QG

HF

ND

HF＋0.25%QG

HF＋1.25%QG

20

25

30

35

40

45

0 10 20 30 40 50 60 70 80 90

Day

Body

weig

ht

(g)

#

#

####

#

####

##

##

##

#####

#

Feed efficiency

Exp.2：In vitro study (3T3-L1 adipocyte) Exp.3：Clinical study

Histological evaluation (Epididymal adipose tissue)

0

0.5

1

1.5

2

2.5

3

3.5

4

4.5

EpididymalPerirenal & retroperitoneal Mesenteric Total Liver

Tis

sue w

eig

ht (

g)

Visceral adipose tissue

##

##：P<0.01 vs. HF#：P<0.05 vs. HF

HF＋0.05%QG

HF

ND

HF＋0.25%QG

HF＋1.25%QG

## ## ##

##

##

##

#

0

0.5

1

1.5

2

2.5

3

3.5

4

4.5

EpididymalPerirenal & retroperitoneal Mesenteric Total Liver

Tis

sue w

eig

ht (

g)

Visceral adipose tissue

##

##：P<0.01 vs. HF#：P<0.05 vs. HF

HF＋0.05%QG

HF

ND

HF＋0.25%QG

HF＋1.25%QG

## ## ##

##

##

##

#

Biochemical parameters

TG

0

10

20

30

40

50

60

mg/dL

NEFA

0.0

0.2

0.4

0.6

0.8

1.0

1.2

1.4

mEq/L

Glucose

50

100

150

200

250

mg/dL

05101520

25303540

Leptin

ng/m

L

#

0

0.5

1

1.5

2

2.5

3 Adiponectin

ng/m

L # ##

HDL

0

20

40

60

80

100

120

140

160

mg/dL

####

LDL

0

5

10

15

20

25

30

mg/dL

0

#

020406080

100120140160180

mg/dL

TC

###

##

##：P<0.01 vs. HF,#：P<0.05 vs. HF

HF＋0.05%QG

HF

ND

HF＋0.25%QG

HF＋1.25%QG

TG

0

10

20

30

40

50

60

mg/dL

TG

0

10

20

30

40

50

60

mg/dL

NEFA

0.0

0.2

0.4

0.6

0.8

1.0

1.2

1.4

mEq/L

NEFA

0.0

0.2

0.4

0.6

0.8

1.0

1.2

1.4

mEq/L

Glucose

50

100

150

200

250

mg/dL

Glucose

50

100

150

200

250

mg/dL

05101520

25303540

Leptin

ng/m

L

#

05101520

25303540

Leptin

ng/m

L

#

0

0.5

1

1.5

2

2.5

3 Adiponectin

ng/m

L # ##

0

0.5

1

1.5

2

2.5

3 Adiponectin

ng/m

L # ##

HDL

0

20

40

60

80

100

120

140

160

mg/dL

####

HDL

0

20

40

60

80

100

120

140

160

mg/dL

####

LDL

0

5

10

15

20

25

30

mg/dL

0

LDL

0

5

10

15

20

25

30

mg/dL

0

#

020406080

100120140160180

mg/dL

TC

###

###

020406080

100120140160180

mg/dL

TC

###

##

##：P<0.01 vs. HF,#：P<0.05 vs. HF

HF＋0.05%QG

HF

ND

HF＋0.25%QG

HF＋1.25%QG

Proposed mechanism of the lipolytic-pathway activation with

Quercetin

Protocol(Lipolysis assesment)

0

100

200

300

400

500

600

[% o

f control]

NAd

Q

－ －

－ － 1μM

＋ ＋ ＋ ＋

3μM 10μM 10μM

##

##

##

0

100

200

300

400

500

[% o

f control]

NAd

Q

－ －

－ － 1μM

＋ ＋ ＋ ＋

3μM 10μM 10μM

##

##

##

##：P<0.01 vs. NAd(+)/Q(-)

0

100

200

300

400

500

600

[% o

f control]

NAd

Q

－ －

－ － 1μM

＋ ＋ ＋ ＋

3μM 10μM 10μM

##

##

##

0

100

200

300

400

500

[% o

f control]

NAd

Q

－ －

－ － 1μM

＋ ＋ ＋ ＋

3μM 10μM 10μM

##

##

##

##：P<0.01 vs. NAd(+)/Q(-)

Released NEFA

Released Glycerol

Phosphorylated-HSL protein level Protocol

Subject backgroud

Dietary intakes for trial period

Start（0w）For trial period

(～12w)

Placebo 1946 ± 44 1917 ± 43

QG-containing

1950 ± 50 1896 ± 37

Placebo 71.4 ± 2.0 68.5 ± 1.8

QG-containing

70.9 ± 1.9 69.7 ± 1.8

Placebo 60.8 ± 2.0 61.5 ± 1.8

QG-containing

60.7 ± 2.3 61.3 ± 1.6

Placebo 264.9 ± 5.8 260.4 ± 5.8

QG-containing

259.7 ± 6.4 256.0 ± 5.4

Fatintake

(g)

Carbohydrateintake

(g)

Energyintake

(kcal)

Proteinintake

(g)

Time-cource change in fat area

Total Visceral Subcutaneous

Placebo beverage QG-containing beverage

（n = 98） （n = 97）

Sex (N)

Age (Y) 43 ± 1 42 ± 1

Height (cm) 163.2 ± 0.8 163.6 ± 0.9

Body weight (kg) 72.5 ± 0.8 72.2 ± 0.9

BMI (kg/m2) 27.2 ± 0.2 26.9 ± 0.2

Waist circumference (cm) 91.4 ± 0.4 91.8 ± 0.5

Hip circumference (cm) 98.3 ± 0.4 98.2 ± 0.4

Subcutaneous fat thickness（arm） (cm) 2.8 ± 0.1 2.7 ± 0.1

Subcutaneous fat thickness（back） (cm) 3.1 ± 0.1 3.1 ± 0.1

Systolic blood pressure (mmHg) 126 ± 1 126 ± 1

Diastolic blood pressure (mmHg) 78 ± 1 78 ± 1

Body fat mass (g) 23657 ± 447 24068 ± 495

Lean body mass (g) 45715 ± 858 45016 ± 838

Total fat area (cm2) 303.1 ± 5.4 305.7 ± 6.9

Visceral fat area (cm2) 94.9 ± 3.5 97.1 ± 3.4

Subcutaneous fat area (cm2) 208.2 ± 5.4 208.5 ± 6.0

♂：50　♀：48 ♂：50　♀：47

Change in anthropometric values

【Summary of Exp.2】Quecetin accelerated the NEFA and glycerol release and phosphorylation of HSL in dose-dependent manner in 3T3-L1 adipocytes,

【Summary of Exp.3】In a clinical study, daily quercetin glucoside intake for 12w resulted in significant reduction of visceral fat mass in healthy obese people and they had experienced no adverse effects.

【Summary of Exp.1】 Supplementation of quercetin glucosides in diet-induced obese mice resulted in reduced body weight gain, and significant reduction of adipose tissue mass and hypertrophy.

Initial BW (g) 21.4 ± 0.27 21.5 ± 0.23 21.1 ± 0.25 20.95 ± 0.22 21.7 ±0.37

Final BW (g)32.2 ± 0.55 ## 39.8 ± 1.09 38.4 ± 1.07 37.10 ±0.87 35.8 ± 0.97 #

⊿BW(g/10heads/88days)

Food intake(g/10heads/88days)

Energy intake(kcal/10heads/88days)

Feed efficiency(⊿BW／energy intake)

0.01127

20547 16322 21408 19182 12549

0.00526 0.01124 0.00804 0.00842

108.0 183.50 172.20

5676 3450.8 4526.1 4064 2687.1

161.50 141.40

ND HFHF+

0.05%QGHF+

0.25%QGHF+

1.25%QG

##:p<0.05, ＃；p<0.01 vs. HF (One-way ANOVA)

Initial BW (g) 21.4 ± 0.27 21.5 ± 0.23 21.1 ± 0.25 20.95 ± 0.22 21.7 ±0.37

Final BW (g)32.2 ± 0.55 ## 39.8 ± 1.09 38.4 ± 1.07 37.10 ±0.87 35.8 ± 0.97 #

⊿BW(g/10heads/88days)

Food intake(g/10heads/88days)

Energy intake(kcal/10heads/88days)

Feed efficiency(⊿BW／energy intake)

0.01127

20547 16322 21408 19182 12549

0.00526 0.01124 0.00804 0.00842

108.0 183.50 172.20

5676 3450.8 4526.1 4064 2687.1

161.50 141.40

ND HFHF+

0.05%QGHF+

0.25%QGHF+

1.25%QG

Initial BW (g) 21.4 ± 0.27 21.5 ± 0.23 21.1 ± 0.25 20.95 ± 0.22 21.7 ±0.37

Final BW (g)32.2 ± 0.55 ## 39.8 ± 1.09 38.4 ± 1.07 37.10 ±0.87 35.8 ± 0.97 #

⊿BW(g/10heads/88days)

Food intake(g/10heads/88days)

Energy intake(kcal/10heads/88days)

Feed efficiency(⊿BW／energy intake)

0.01127

20547 16322 21408 19182 12549

0.00526 0.01124 0.00804 0.00842

108.0 183.50 172.20

5676 3450.8 4526.1 4064 2687.1

161.50 141.40

ND HFHF+

0.05%QGHF+

0.25%QGHF+

1.25%QG

##:p<0.05, ＃；p<0.01 vs. HF (One-way ANOVA)

Body weight

Tissue weight

【Evaluation】●Body weight, general state, food intake●Liver weight, visceral fat mass●Histlogical evaluation(Epididymal adipose tissue)●Blood biochemical parameter（TG,Cho,NEFA,Glc）、Liver TG contents●Adipocytokine

High Fat ＋ 0.05% Quercetin glucosides diet（HF＋0.05%QG）

High Fat Diet （HF）Normal Diet（ND）

ad. Lib. for 3 monthsC57BL/6J mice、♂

6weeks old

10mice/group

High Fat ＋ 0.25% Quercetin glucosides diet（HF＋0.25%QG）

High Fat ＋ 1.25% Quercetin glucosides diet（HF＋1.25%QG）

【Evaluation】●Body weight, general state, food intake●Liver weight, visceral fat mass●Histlogical evaluation(Epididymal adipose tissue)●Blood biochemical parameter（TG,Cho,NEFA,Glc）、Liver TG contents●Adipocytokine

High Fat ＋ 0.05% Quercetin glucosides diet（HF＋0.05%QG）

High Fat Diet （HF）Normal Diet（ND）

ad. Lib. for 3 monthsC57BL/6J mice、♂

6weeks old

10mice/group

High Fat ＋ 0.25% Quercetin glucosides diet（HF＋0.25%QG）

High Fat ＋ 1.25% Quercetin glucosides diet（HF＋1.25%QG）

Activated pathway with

Quercetin

Activated pathway with

Quercetin

QG(275mg※)-containing bevarage 350mL/1bottle/day

For 12w

●Study Design: A double-blind placebo-controlled study●Subjects: 200 overweight and obese subjects(BMI24～35kg/m2)●Sex: ♂102, ♀98

N=100

Placebo bevarage 350mL/1bottle/day N=100

【Evaluation】●Body weight●Abdominal total fat area (CT scan with CT-W450, hitachi medico Ltd., Japan)●Body composition (DEXA(Dual Ebergy X-ray absorptiometry) methods with ProdigyTM, GE Healthcare, USA)●Blood biological parameter

QG(275mg※)-containing bevarage 350mL/1bottle/day

For 12w

●Study Design: A double-blind placebo-controlled study●Subjects: 200 overweight and obese subjects(BMI24～35kg/m2)●Sex: ♂102, ♀98

N=100

Placebo bevarage 350mL/1bottle/day N=100

【Evaluation】●Body weight●Abdominal total fat area (CT scan with CT-W450, hitachi medico Ltd., Japan)●Body composition (DEXA(Dual Ebergy X-ray absorptiometry) methods with ProdigyTM, GE Healthcare, USA)●Blood biological parameter

0-4w 0-8w 0-12w

Placebo -0.2 ± 0.1* -0.2 ± 0.1 -0.4 ± 0.2*

QG-containing

-0.1 ± 0.1 -0.3 ± 0.2 -0.4 ± 0.2*

Placebo -0.1 ± 0.0* -0.1 ± 0.0 -0.1 ± 0.1*

QG-containing

0.0 ± 0.0 -0.1 ± 0.1 -0.1 ± 0.1*

Placebo -0.2 ± 0.1* -0.3 ± 0.1* -0.4 ± 0.2*

QG-containing

-0.3 ± 0.1** -0.8 ± 0.1***,## -1.4 ± 0.2***,###

Placebo -0.1 ± 0.1 0.0 ± 0.1 -0.1 ± 0.1

QG-containing

-0.1 ± 0.1 -0.4 ± 0.1**,## -0.6 ± 0.1***,##

Placebo 0.0 ± 0.0 0.0 ± 0.0* -0.1 ± 0.0***

QG-containing

0.0 ± 0.0 0.0 ± 0.0*** -0.1 ± 0.0***

Placebo 0.0 ± 0.0*** 0.0 ± 0.0*** -0.1 ± 0.0***

QG-containing

0.0 ± 0.0** -0.1 ± 0.0*** -0.1 ± 0.0***,##

Placebo -17 ± 95 38 ± 114 -130 ± 141

QG-containing

-140 ± 96 -351 ± 150*,# -301 ± 198

Placebo -189 ± 103 -68 ± 94 -238 ± 97*

QG-containing

-17 ± 89 133 ± 91 -80 ± 91

Waist circumference (cm)

Hip circumference (cm)

Body weight (kg)

BMI (kg/m2)

Subcutaneous fat thickness（arm） (cm)

Lean body mass (g)

Body fat mass (g)

Subcutaneous fat thickness（back） (cm)

###：P<0.001, ##:P<0.01, #:P<0.05 vs. placebo、***：P<0.001, **：P<0.01, *：P<0.05 vs. 0w

0-4w 0-8w 0-12w

Placebo -0.2 ± 0.1* -0.2 ± 0.1 -0.4 ± 0.2*

QG-containing

-0.1 ± 0.1 -0.3 ± 0.2 -0.4 ± 0.2*

Placebo -0.1 ± 0.0* -0.1 ± 0.0 -0.1 ± 0.1*

QG-containing

0.0 ± 0.0 -0.1 ± 0.1 -0.1 ± 0.1*

Placebo -0.2 ± 0.1* -0.3 ± 0.1* -0.4 ± 0.2*

QG-containing

-0.3 ± 0.1** -0.8 ± 0.1***,## -1.4 ± 0.2***,###

Placebo -0.1 ± 0.1 0.0 ± 0.1 -0.1 ± 0.1

QG-containing

-0.1 ± 0.1 -0.4 ± 0.1**,## -0.6 ± 0.1***,##

Placebo 0.0 ± 0.0 0.0 ± 0.0* -0.1 ± 0.0***

QG-containing

0.0 ± 0.0 0.0 ± 0.0*** -0.1 ± 0.0***

Placebo 0.0 ± 0.0*** 0.0 ± 0.0*** -0.1 ± 0.0***

QG-containing

0.0 ± 0.0** -0.1 ± 0.0*** -0.1 ± 0.0***,##

Placebo -17 ± 95 38 ± 114 -130 ± 141

QG-containing

-140 ± 96 -351 ± 150*,# -301 ± 198

Placebo -189 ± 103 -68 ± 94 -238 ± 97*

QG-containing

-17 ± 89 133 ± 91 -80 ± 91

Waist circumference (cm)

Hip circumference (cm)

Body weight (kg)

BMI (kg/m2)

Subcutaneous fat thickness（arm） (cm)

Lean body mass (g)

Body fat mass (g)

Subcutaneous fat thickness（back） (cm)

###：P<0.001, ##:P<0.01, #:P<0.05 vs. placebo、***：P<0.001, **：P<0.01, *：P<0.05 vs. 0w

●QG

○Placebo

●QG

○Placebo

●QG

○Placebo

Glycerol and NEFA quantification・・・Index of lipolysis

Cells; 3T3-L1 pre-adipocytes (3.5~5×104cells/well)

maintained in DMEM supplement with 10% CSuntil super confluent

• 0.25µM dexamethasone• 500µM 3-isobutyl-1-methylxanthine• 10µg/mL insulin

maintained in DMEM supplement with 10% FBS+ 5µg/mL insulin for 14 days

• 0.1µM noradrenaline• 5µg/mL insulin

• Quercetin

Western blot analysis on phosphorylated-HSL ※

・・・Index of lipolytic pathway activation

※Phosphorylated hormone-sensitive lipase

1. Control2. 0.1 µM L-Noradrenaline3. 0.1 µM L-N+3 µM Quercetin4. 0.1 µM L-N+10 µM Quercetin5. 0.1 µM L-N+10 µM Quercetin+ 10 µM Propranolol6. 10 µM Isoproterenol7. 3 µM Quercetin8. 10 µM Quercetin

Phospho-HSL (Ser660)