Elise Saunier Bionovo, Inc. NAMS 2009 Identification of Tissue-Selective ER agonists from Plants.
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Transcript of Elise Saunier Bionovo, Inc. NAMS 2009 Identification of Tissue-Selective ER agonists from Plants.
Elise Saunier
Bionovo, Inc. NAMS 2009
Identification of Tissue-Selective ERIdentification of Tissue-Selective ER agonists from Plantsagonists from Plants
Endocrine changes during the menopausal transition, rather than the aging Endocrine changes during the menopausal transition, rather than the aging process, are related to changes in body weight and fat distribution.process, are related to changes in body weight and fat distribution.
In age-matched women: higher percentage of fat tissue in peri- and post-In age-matched women: higher percentage of fat tissue in peri- and post-menopausal women (p<0.001) compare to premenopausal and menopausal women (p<0.001) compare to premenopausal and postmenopausal + HRT womenpostmenopausal + HRT women
Perimenopausal and postmenopausal women show a shift to a central, Perimenopausal and postmenopausal women show a shift to a central, android fat distribution. Regional distribution shows higher fat distribution android fat distribution. Regional distribution shows higher fat distribution in the trunk (p<0.001) and lower fat in the legs (p<0.05) for the in the trunk (p<0.001) and lower fat in the legs (p<0.05) for the postmenopausal womenpostmenopausal women
Overweight and obesity are associated with increased morbidity and Overweight and obesity are associated with increased morbidity and mortality, with central distribution of body fat recognized as an independent mortality, with central distribution of body fat recognized as an independent predictor of cardiovascular disease in womenpredictor of cardiovascular disease in women
Can be counteracted by HRTCan be counteracted by HRT
Weight Gain during MenopauseWeight Gain during Menopause
2
Fat redistribution during menopauseFat redistribution during menopause
*p<0.05 (vs. premenopause value); **p<0.01 (vs premenopause value)3
Genazzani & Gambacciani studied the modifications of body weight and fat Genazzani & Gambacciani studied the modifications of body weight and fat distribution from 2529 womendistribution from 2529 women
Mammary GlandMammary GlandUterusUterus
Breast CancerBreast CancerEndometrial CancerEndometrial Cancer
Adipose TissueAdipose Tissue
Decreased ObesityDecreased ObesityDecreased Metabolic SyndromeDecreased Metabolic Syndrome
ER
Estrogen
in HT
Estrogen
in HT
ER
Estrogen
in HT
Estrogens in HT are not Tissue SelectiveEstrogens in HT are not Tissue Selective
4
Adipose TissueAdipose Tissue
Decreased ObesityDecreased ObesityDecreased Metabolic SyndromeDecreased Metabolic Syndrome
ER
Tissue Selective ERTissue Selective ERαα-estrogens-estrogens
5
Tissue Selective
ERα Agonist
Tissue Selective
ERα Agonist
Mammary GlandMammary GlandUterusUterus
Breast CancerBreast CancerEndometrial CancerEndometrial Cancer
ER
Tissue Selective
ERα Agonist
SAFERSAFERSAFERSAFER
To identify tissue selective ERTo identify tissue selective ER agonists in plants agonists in plants
MethodsMethods
1.1. Screened over 50 plant extracts for activation of an ERE reporter Screened over 50 plant extracts for activation of an ERE reporter genegene
2.2. Selected two plant extracts for testing in mice by gavage:Selected two plant extracts for testing in mice by gavage:
#3 from Radix Glycyrrhiza#3 from Radix Glycyrrhiza#39 from Radix Pueraria#39 from Radix Pueraria
3.3. Ovariectomized mice were pre-fed for 50 days with HFD before Ovariectomized mice were pre-fed for 50 days with HFD before started treatment with E2, #3 and #39 for 42 daysstarted treatment with E2, #3 and #39 for 42 days
4.4. Measured weights of body, abdominal fat, MG and uterusMeasured weights of body, abdominal fat, MG and uterus
5.5. Determine gene expression profiles in abdominal fat, MG and Determine gene expression profiles in abdominal fat, MG and uterusuterus
GoalGoal
6
7
Plant extract #3 and #39 induce ERE-Luciferase reporter expression in vitroPlant extract #3 and #39 induce ERE-Luciferase reporter expression in vitro
Plant Extracts have ERα ActivityPlant Extracts have ERα Activity
*p<0.05 vs. control
ERE-ERα tk-Luc
0
1
2
3
4
10-8 0.2 0.5 1 2.5 10 20 100 0.2 0.5 1 2.5 10 20 100
RL
UControlE2#3#39
**
**
***
*****
EC50 = 16 μg/mlEC50 = 16 μg/ml EC50 = 5.5 μg/mlEC50 = 5.5 μg/ml
4.8
5.0
5.2
5.4
5.6
5.8
0.1 1 10 100 1000
concentration (ug/ml)
4.8
5.0
5.2
5.4
5.6
5.8
0.1 1 10 100 1000
concentration (ug/ml)
Binding assay
8
Consumption of a high fat diet increases body weight in miceConsumption of a high fat diet increases body weight in mice
E2 administration reverses the weight gainE2 administration reverses the weight gain
Plant extract #3 and #39 also reverse weight gainPlant extract #3 and #39 also reverse weight gain
Decrease in Body Weight by the Plant ExtractsDecrease in Body Weight by the Plant Extracts
-30
-20
-10
0
10
7 14 21 28 35 42
days in HFD
% w
eig
ht
chan
ge
vehicle
E2
3
39
***
**
*
**
*
**
*
*
*
*
**
*p<0.05 vs. control
Abdominal fat
0
0.5
1
1.5
2
2.5w
eig
ht
(g) vehicle
E2
3
39
* *
*
9
#3 and #39 mimic estradiol’s effect on the abdominal fat#3 and #39 mimic estradiol’s effect on the abdominal fat
Plant Estrogens Decrease Abdominal FatPlant Estrogens Decrease Abdominal Fat
Abdominal FatAbdominal FatAbdominal FatAbdominal Fat
*p<0.05 vs. control
Uterus
0
100
200
300
wei
gh
t (m
g) vehicle
E2
3
39
*
10
Estradiol treatment induces a massive increase in uterus weightEstradiol treatment induces a massive increase in uterus weight
#3 and #39 do not mimic estradiol’s effect on the uterus#3 and #39 do not mimic estradiol’s effect on the uterus
Uterus Weight is not Increased by Plant ExtractsUterus Weight is not Increased by Plant Extracts
UterusUterusUterusUterus
*p<0.05 vs. control
Mammary gland
0
0.5
1
1.5
2
wei
gh
t (g
) vehicle
E2
3
39
**
*
11
Estradiol reduces the mammary gland weight but induces Estradiol reduces the mammary gland weight but induces
development of ducts (elongation, branching)development of ducts (elongation, branching)
Mammary gland is not affected by the Plant Mammary gland is not affected by the Plant ExtractsExtracts
Mammary GlandMammary GlandMammary GlandMammary Gland controlcontrol
3399
33
EE22
While the weight of the mammary gland is diminished by the plant While the weight of the mammary gland is diminished by the plant
extracts, the ducts do not develop.extracts, the ducts do not develop.
*p<0.05 vs. control
12
#3 and #39 mimic estradiol’s effect on the abdominal fat#3 and #39 mimic estradiol’s effect on the abdominal fat
#3 and #39 DO NOT mimic estradiol’s effect on the uterus#3 and #39 DO NOT mimic estradiol’s effect on the uterus
Plant Extracts have Tissue Specific Plant Extracts have Tissue Specific Estrogenic EffectsEstrogenic Effects
Abdominal FatAbdominal FatAbdominal FatAbdominal Fat
UterusUterusUterusUterus
Mammary GlandMammary GlandMammary GlandMammary Gland
To understand molecular basis for tissue specific effectsTo understand molecular basis for tissue specific effects
gene expression profile by microarraygene expression profile by microarray
13
Compared to estradiol, #3 and #39 regulate a comparable number of genes in Compared to estradiol, #3 and #39 regulate a comparable number of genes in
the abdominal fatthe abdominal fat
Gene regulation by the Plant Extracts is Tissue Gene regulation by the Plant Extracts is Tissue SpecificSpecific
-800
-400
0
400
800
E2 #3 #39
-400
-200
0
200
400
E2 #3 #39
-1500-1000
-5000
5001000
1500
E2 #3 #39
Mammary GlandMammary GlandMammary GlandMammary GlandUterusUterusUterusUterus
Abdominal FatAbdominal FatAbdominal FatAbdominal Fat
However, little or no genes are induced or repressed in the Uterus and MG, However, little or no genes are induced or repressed in the Uterus and MG,
when compare to E2 treatment.when compare to E2 treatment.
14
847
E2E2
#3#3
239239
4848
413413
338
#39#39
45 349
Mammary GlandMammary GlandMammary GlandMammary Gland
469
00
1012
E2E2 #39#39
#3#3
Tissue Specific Activity of Plant EstrogensTissue Specific Activity of Plant Estrogens
1980
#3#3
8181
11112112
2525
167 304
E2E2#39#39
UterusUterusUterusUterus
Abdominal FatAbdominal FatAbdominal FatAbdominal FatE2, #3 and 39 regulate E2, #3 and 39 regulate genes in the fatgenes in the fat
E2 is the major gene E2 is the major gene regulator in the uterus regulator in the uterus
and the mammary glandand the mammary gland
158 Inflammatory 158 Inflammatory genes repressed by genes repressed by E2, #3 and #39E2, #3 and #39
Mammary GlandMammary GlandUterusUterus
Breast CancerBreast CancerEndometrial CancerEndometrial Cancer
Adipose TissueAdipose Tissue
Decreased ObesityDecreased ObesityDecreased Metabolic SyndromeDecreased Metabolic Syndrome
ER
Estrogen
in HT
Estrogen
in HT
ER
Estrogen
in HT
Estrogens in HT are not Tissue SelectiveEstrogens in HT are not Tissue Selective
15
E2E2
669
FatFat
MGMG
7373 99 79
228
UterusUterus
338 1749
modifies gene expression in all modifies gene expression in all
3 tissues3 tissues
Adipose TissueAdipose Tissue
Decreased ObesityDecreased ObesityDecreased Metabolic SyndromeDecreased Metabolic Syndrome
ER
Tissue Selective ERTissue Selective ERαα-estrogens-estrogens
16
Tissue Selective
ERα Agonist
Tissue Selective
ERα Agonist #3#3MGMG
190190 848848 3939
00
FatFat UterusUterus
#39#39
734
MGMG
306
66
358
16
FatFat UterusUterus
Plant Estrogens
#3 and #39 have one major target #3 and #39 have one major target abdominal FAT = GOOD NEWSabdominal FAT = GOOD NEWS
Much less or no effect on MG and Much less or no effect on MG and UterusUterus
•Plant extracts mimic the beneficial effect of Plant extracts mimic the beneficial effect of estradiol on body weight and repression of estradiol on body weight and repression of inflammatory genesinflammatory genes
•Plant extracts have little or no effect on the Plant extracts have little or no effect on the mammary gland and uterusmammary gland and uterus
•Animal data support the goal to use these Animal data support the goal to use these plants in clinical trials to safely treat obesity in plants in clinical trials to safely treat obesity in postmenopausal womenpostmenopausal women
•These studies demonstrate that it is possible These studies demonstrate that it is possible to discover tissue selective ERto discover tissue selective ER agonists. Might agonists. Might have a more favorable clinical profile than have a more favorable clinical profile than estrogens used currently in classic HT for estrogens used currently in classic HT for menopausal symptomsmenopausal symptoms
SummarySummary