ampk activation by "compound 13"
TRANSCRIPT
MB7264 and “Compound 13” In vitro and vivo selective activation of
the AMPK 𝛼1 isoform
Jorge E Gómez-Galeno
A presentation based on:
A Potent and Selective AMPK Activator That Inhibits de Novo Lipogenesis
Jorge E. Gómez-Galeno, Qun Dang, Thanh H. Nguyen, Serge H. Boyer, Matthew P. Grote, Zhili Sun, Mingwei Chen, William A. Craigo, Paul D. van Poelje, Deidre A. MacKenna, Edward E. Cable, Paul A. Rolzin, Patricia D. Finn, Bert Chi, David L. Linemeyer, Scott J. Hecker, and Mark D. Erion
ACS Med. Chem. Lett., 2010, 1 (9), pp 478–482 DOI: 10.1021/ml100143q
AMP-activated Protein Kinase (AMPK)
✤ Widely distributed heterotrimeric ser/thr kinase:
✤ Catalytic alpha subunit, regulatory beta and gamma subunits
✤ Multiple isoforms (𝛼1, 𝛼2, 𝛽1, 𝛽2, 𝛾1, 𝛾2, 𝛾3)
✤ Metabolite sensor: sensitive to AMP/ATP ratio
✤ Activated during exercise, nutrient deprivation, hipoxia/ischemia
AMPK -- Metabolic Effects
✤ Expected effects of a liver targeted AMPK activator
Decreased fatty acid and cholesterol production
Decreased triglyceride accumulation
Increased fatty acid oxidation
http://www.huffingtonpost.com/paul-spector-md/exercise-in-a-bottle-nest_b_6221922.html
AMPK activators
N
NN
NNH2
O
OHOH
OPHOOH
O
O
NH2N
NNH2
O
OHOH
OR
R = H: AICAR R = PO3H2: ZMP
NH
S
HO
OH N
O
A-769662
MB7264
• No activity at 100µM against FBPase, 5’-nucleotidase, AMP deaminase, Adenosine Kinase, or Glycogen Phosphorylase
• No hits or cross-reactivity at 10 μM against 64 targets (MDS lead-profiling screen)
Liver AMPK Activation
Concentration (µM)
0.001 0.01 0.1 1 10 100
AM
PK
Act
ivat
ion
(V/V
0)
1.0
1.5
2.0
2.5
3.0
3.5
MB07264
AMP
EC50 = 7.6 nM
EC50 = 6.3 µM
Human
Liver AMPK Activation
Concentration (µM)
0.001 0.01 0.1 1 10 100
AM
PK
Act
ivat
ion
(V/V
0)
1.0
1.5
2.0
2.5
3.0
3.5
MB07264
AMP
EC50 = 7.6 nM
EC50 = 6.3 µM
Human
Concentration (µM)
0.001 0.01 0.1 1 10 100
AM
PK
Act
ivat
ion
(V/V
0)1
2
3
4
5
MB07264
AMP
EC50 = 20 nM
EC50 = 5.7 µM
Rat
Species-dependent Isoform Activity
Species α1 α2MB7264
Max Activation
Human >95% <5% 115%Rat 50% 50% 43%
Mouse 50% 50% 54%Guinea Pig 95% 5% 96%
MB7264
MB7264
O PO3Bn2N
O PNO
O
H2,Pd(OH)2/C
OO
OHChx2NHEthanol Chx2NH2
O
BnO2
O PO3R2O
HR = Et
R = H
R = CH2Ph
O PO3Bn2HON
XX = H
X = ClTMSBr, CH2Cl2
NNHChx
Chx OBn
BnO
Et3N, CH2Cl2
NH2OH-HCl
NaOAc, THF: H2O
NCS, DMF
MB7264 - properties
ONO
HO
PO3Et2ONO
HO
PO3H2
pKa 3.32, 3.93, 6.94
logD7.4 -4.3
pKa 3.23
Esterase Sensitive Prodrugs
O PO3H2
O
H O PO
H
OO
O
O2
I O
O
Et3N, CH3CN
(1) NH2OH-HCl, NaOAc
(2) NCS, DMF
O PHON
Cl
OO
O
O O PN O
O
O
O2
O
O2
O O
OO
Et3N, CH2Cl2
Intracellular concentration of MB7264*
Concentration Intracellular MB7264 (nmol/g)
100 µM < 55
10 µM
100 µM
254 ± 104
2151 ± 712O P
N OO
O
O2
O
O
O PNO
O
OO
OH
Chx2NH2 2
* Plated rat hepatocytes
Inhibition of de novo lipogenesis*
O PN O
O
O
O2
O
OR
R EC50 (nM) R EC50 (nM)
N/A > 10,000 OEt 42
t-Bu 100 O-(i-Pr) 27
i-Pr 20 OCH2Ph 609
Cyclopentyl 30 OC(CH3)2Et 390
* Plated rat hepatocytes
Inhibition of de novo lipogenesis*
O PN O
O
O
O2
O
OR
R In vivo DNL inhibition
R In vivo DNL inhibition
N/A ND OEt 73%
t-Bu 65% O-(i-Pr) 73%
i-Pr 78% OCH2Ph 34%
Cyclopentyl ND OC(CH3)2Et ND
*C57Bl/6 mice, 30 mg/kg ip
“Compound 13”
AMPK Activators - Summary
• MB07264 is a direct, selective, potent activator of AMPK that activates α1−containing complexes.
• In hepatocytes, a prodrug of MB07264 regulates several expected downstream targets: – Increases ACC phosphorylation
– Decreases malonyl CoA levels
– Inhibits de novo lipogenesis
• Prodrugs of MB7264 inhibit de novo lipogenesis in mice and rats (data not shown) and are preferentially distributed into liver tissues
Other studies on our MB7264 prodrug
http://www.huffingtonpost.com/paul-spector-md/exercise-in-a-bottle-nest_b_6221922.html
http://www.huffingtonpost.com/paul-spector-md/exercise-in-a-bottle-nest_b_6221922.html
http://www.huffingtonpost.com/paul-spector-md/exercise-in-a-bottle-nest_b_6221922.html
Studies confirming that: • MB7264 has 𝛼1 selectivity
and • its prodrugs inhibit liver de
novo lipogenesis
A prodrug of MB7264 in Oncology
Compound 13, an α1-selective small molecule activator of AMPK, potently inhibits melanoma cell proliferation Xueqing Hu, Fangzhen Jiang, Qi Bao, Huan Qian,
Quan Fang, Zheren Shao
Tumor Biology, pp 1-8; First online: 14 August 2015 DOI: 10.1007/s13277-015-3854-8
Compound 13