MHY2013 is a novel PPAR pan-agonist that was shown to have beneficial effects on metabolic disorders in mouse models of obesity and diabetes. It activated all PPAR subtypes and increased fatty acid oxidation and energy expenditure in liver, adipose tissue, and skeletal muscle by upregulating genes involved in these pathways. MHY2013 improved insulin sensitivity, lowered blood triglycerides and fatty acids, and reduced hepatic steatosis in obese mice without affecting food intake or body weight. The compound's metabolic effects were mediated through increased levels of the hormones FGF21 and adiponectin.

1. A novel PPAR pan-agonist, 2-(4-(5,6-
methylenedioxybenzo[d]thiazol-2-yl)-2-methylphenoxy)-2-
methylpropanoic acid (MHY2013) in the treatment of
metabolic disorders
Presenter : Sultan Ullah
PhD Scholar
Professor Moon’s Lab, College of Pharmacy,
Pusan National University, Busan, South Korea.
1
Oral presentation in

2. Contents
Introduction
Results and discussion
Summary
1
2
3
2

3. Symptoms
(1) Excessive weight
(2) Hypertension
(3) High blood glucose level
(4) High level of lipids, TGs
and LDL.
Therapeutic approaches
• PPARs receptors (α, β/δ, γ)
• Selective agonist showed ADRS
• Need for dual and pan agonists
Cardiometabolic complications
• Dyslipidemia
• Type 2 diabetes mellitus (T2DM)
• Nonalcoholic fatty liver disease (NAFLD)Epidemiology
• Affects more than 35% people in the US
and Australia
Metabolic disorders
• Misregulation of energy
production and metabolism
• Obesity is the main cause
1. Introduction
3

4. WAT BAT
• White adipose tissue
• lipid-storage depots
• Brown adipose tissue
• Lipid-storage depots
• Naturally undergoes thermogenesis
(Cold exposure/β-adrenergic
activation))
Browning of WAT
• Thermogenic stimuli Increase beige adipocytes
• Prevents metabolic disorders
• PPARs activation–Prdm16-(FGF21, adiponectin, UCP-1, Cidea, Pgc1α,
Irisin)
• Dyslipidemia
• T2DM
• Insulin resistance
• NAFLD*
1.1. Adipose tissue and metabolic disorders
4
*Non alcoholic fatty liver disease Nature Reviews Endocrinology 13,36–49, (2017)

5. 1.2. PPARs and metabolic disorders
Nature Reviews Endocrinology 13,36–49, (2017)

6. 2. Results and Discussion
6

7. Synthetic route
2.1. Synthesis of MHY2013
7

8. 2.2. Luciferase assay
• YPEN-1 cells
• 3X-PPRE-TK-LUC plasmid
• 10 µM Concentration
• The data are shown as the
mean ± SEM (n = 4) luciferase
activity
PPRE-luc + PPAR
C
onW
Y14643M
H
Y2013M
H
Y1907M
H
Y2014M
H
Y2015M
H
Y2016M
H
Y2062
0
1000000
2.0100 6
3.0100 6
4.0100 6
*
###
###
#
luciferaseactivity
PPRE-luc + PPAR/
C
O
N
G
W
501516M
H
Y
2013M
H
Y
1907M
H
Y
2014M
H
Y
2015M
H
Y
2016M
H
Y
2602
0
100000
200000
300000
400000
500000
**
## #
luciferaseactivity PPRE-luc + PPAR
C
O
N
R
osiglitazoneM
H
Y2013M
H
Y1907M
H
Y2014M
H
Y2015M
H
Y2016M
H
Y2062
0
100000
200000
300000
*
###
luciferaseactivity
8

9. • AutoDock Vina
• Chimera
• LigandScout.
• MHY2013 might directly bind to the
three PPAR subtypes
2.3. In silico binding mode representation
9
PPARα
NAME DOCKING SCORE
WY14643 -7.9
MHY2013 -8.7
PPARβ/δ
NAME DOCKING SCORE
GW501516 -9.7
MHY2013 -9.7
PPARγ
NAME DOCKING SCORE
Rosiglitazone -8.0
MHY2013 -7.7

10. 2.3. In silico binding mode representation
10
PPARα
PPARβ/δ PPARγ

11. 2.4. In vitro and In vivo toxicity
• 3-(4,5-Dimethylthiazol-2-yl)-2,5-
diphenyltetrazolium bromide
(MTT)
• Serum creatinine: renal injury
• Serum aspartate transaminase
(AST) and alanine
transaminase (ALT): liver injury
• 10 μM concentration
• No severe toxicity
C
on
0.1
0.5
1
5
10
0
50
100
150 MTT
MHY2013(M)
Cellviability(%)
db/m
db/db
M
H
Y2013
0.0
0.5
1.0
1.5
Creatinine
Relativetodb/m
db/m
db/db
M
H
Y2013
0
5
10
15
20
25
AST
IU/L
db/m
db/db
M
H
Y2013
0
5
10
15 ALT
IU/L
11

12. 2.5. Transcriptional activity
• PPARs in nucleus
• Immunofluorescence staining
• Western blotting
• qRT-PCR
• mRNA levels of the target
genes
• MHY2013 is a strong PPAR
pan agonist targeting all
PPAR subtypes
PPAR γ
Rosiglitazone(μM) - 10 - -
MHY2013(μM) - - 1 10
TFⅡB
TFⅡB
PPAR α
WY14643(μM) - 10 - -
MHY2013(μM) - - 1 10
GW501516(μM) - 10 - -
MHY2013(μM) - - 1 10
PPAR β/δ
TFⅡB
0
2
4
6
8
10
Acox1 Cpt1 Pdk4 Hmgcs2 Scd1 Acc
**
***
* *
***
Con
MHY2013 1M
MHY2013 10M
PPARs mRNA target genesRelativemRNAexpression
12
PPAR /
C
o
n
M

G
W
501516
10
M

M
H
Y
2013
1
M

M
H
Y
2013
10
0.0
0.5
1.0
1.5
2.0
**
**
Arbitraryunit
PPAR
C
o
n
M

W
Y
14643
10
M

M
H
Y
2013
1
M

M
H
Y
2013
10
0.0
0.5
1.0
1.5
2.0
2.5
**
**
***
Arbitraryunit
PPAR
C
on
M

R
osig
litazon
e
10
M

M
H
Y
2013
1
M

M
H
Y
2013
10
0
1
2
3
*
**
Arbitraryunit
Acox1: Acyl-CoA Oxidase 1
Cpti: Carnitine palmitoyltransferase I
Pdk4: Pyruvate Dehydrogenase Kinase 4
Hmgcs2: 3-hydroxy-3-methylglutaryl-CoA synthase 2
Scd1: Stearoyl-CoA Desaturase
Acc: Acetyl-CoA carboxylase

13. 2.6. Effect on blood profile of TGs and NEFA
• Mouse model db/db and db/dm
• MHY2013 (5 mg/kg/day)
• No differences observed in the
food intake and body weight
• Serum levels of TGs and NEFAs
reduced
Food intake
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
0
2
4
6
8
10 db/m
db/db
MHY2013
days
g
Body weight
3 6 9 12 15 18 21
0
20
40
60
db/m
db/db
MHY2013
days
g
Triglyceride
db/m
db/db
M
H
Y2013
0
20
40
60
80
100 ***
###
mg/dL
NEFA
db/m
db/db
M
H
Y2013
0
200
400
600
**
#
uEq/L
13
(Non esterified fatty acid)

14. Glucose tolerance test
0 15 30 60 120 180
0
200
400
600
800
1000
db/m
db/db
MHY2013
***
*** ***
***
***
***
###
#
###
#
###
##
##
Mins
mg/dL
db/m
db/db
M
H
Y2013
0
50000
100000
150000
***
##
Glucose tolerance test (GTT)
AUC
• The glucose tolerance
test (GTT)
• Reduced the fasting
blood glucose and
serum insulin
2.7. Decrease in insulin sensitivity
14
Fasting Glucose
db/m
db/db
M
H
Y20130
100
200
300
400
500
***
###
mg/dL
Insulin
db/m
db/db
M
H
Y2013
0
1
2
3
4 ***
###
ng/ml

15. 2.8. Effect on liver of obese mice
• Induced phosphorylation of
LKB1
• Liver AMPKα1/2 (Thr172)
phosphorylation increased
• Increased mRNA expression
levels of fatty acid oxidation-
related genes: qRT - PCR
• Increased mRNA expression
levels of FGF21: qRT - PCR
• Increased serum level of
FGF21:Mouse/rat FGF21 Quantkine ELISA Kit.
• Luciferase assay using HepG2
cells
Liver TG
db/m
db/db
M
H
Y
2013
0
5
10
15
***
#
mg/100mgprotein
AMPK
FGF21: Fibroblast growth factor
db/m db/db MHY2013
p-AMPKp-LKB1
db/m db/db MHY2013
β-actin
0
2
4
6
8
Acox1 Cpt1 Hmgcs2 Pdk4
###
*
##
db/m
db/db
MHY2013
Mitochondria FA -oxidation
RelativemRNAexpression
Fgf21 (liver)db/m
db/db
M
H
Y2013
0.0
0.5
1.0
1.5 ###
RelativemRNAexpression
serum FGF21
db/m
db/db
M
H
Y2013
0
100
200
300
400
500 ##
pg/ml
FGF21-luc
C
on
M
W
Y14643
10
M
M
H
Y2013
10
0
100000
200000
300000 ***
***
luciferaseactivity
15
(HepG2 cells)
p-AMPK

16. 2.9. Effect on adipose tissue of obese mice
• Dose-dependently increased the
mRNA levels of browning markers
• Increased expression level of
adiponectin in adipose tissue.
• Increased serum adiponectin level
• Increased mRNA expression of
adiponectin in 3T3-L1 cells
0
2
4
6
8 db/m
db/db
MHY2013
#
*
*
##
Ucp1 Cidea Cd137Pgc1 Slc27a1
Adipose tissue
RelativemRNAexpression
0
2
4
6
Ucp1 Cidea Cd137Pgc1
Con
MHY2013 1M
MHY2013 10M
Slc27a1
&
&
&&&
3T3-L1 adipocytes
RelativemRNAexpression
Serum adiponectin
db/m
db/db
M
H
Y2013
0
5
10
15
**
#
ng/ml
Adiponectin Adipose tissue
db/m
db/db
M
H
Y2013
0
1
2
3
4
5
**
RelativemRNAexpression
Adiponectin (3T3-L1)
C
on
M
H
Y2013
1uM
M
H
Y2013
10uM
0
1
2
3
***
RelativemRNAexpression
16
Ucp1: Uncoupling Protein 1
Cidea: Cell death activator CIDE-A
Pgc1α: PPARgamma coactivator 1-alpha
CD137: is a member of the tumor necrosis factor
Slc27a1: Long-chain fatty acid transport protein 1

17. 0
2
4
6
8
Acox1 Cpt1 Hmgcs2 Pdk4
###
*
##
db/m
db/db
MHY2013
Mitochondria FA -oxidation
RelativemRNAexpression
0
10
20
30
40
50
***
#
*** #
db/m
db/db
MHY2013
Mcp1 Tnf
Inflammatory genes
RelativemRNAexpression
• The mRNA expression levels of fatty
acid oxidation-related genes
2.9. Effect on adipose tissue of obese mice
Mcp1: Monocyte Chemotactic Protein 1
Tnfa: Tumor necrosis factor alpha
• MHY2013 also reduced the mRNA
expression levels of inflammatory genes

18. 2.10.Effect on skeletal muscle of obese mice
• Increased expression levels of fatty acid
oxidation-related genes
IRISIN
db/m
db/db
M
H
Y2013
0.0
0.5
1.0
1.5
2.0
##
Relativeexpression
Mitochondria FA -oxidation
0
1
2
3
4
5
ACOX1 CPT1 HMGCS2 PDK4
db/m
db/db
MHY2013
#
##
##
Relativeexpression
18
• Increased irisin, a recently identified myokine that
improves obesity related metabolic syndrome

19. 3. Summary
19

20. Summary
Browning Insulin sensitivity Dyslipidemia
Liver
Adipose
tissue
Blood FGF21
FGF21
FGF21FGF21 Adiponectin
Adiponectin
Adiponectin
MHY2013
PPARα
PPARβ/δ
FGF21
β- oxidation
P-AMPK
MHY2013
PPARγ
Adiponectin
FGF21
PGC1α
UCP1
Hepatic steatosisHepatic steatosis

21. Thank you for your attention.
21