'Lo último en obesidad'. Este es el título del Simposio Internacional que organizamos en la Fundación Ramón Areces los días 1 y 2 de diciembre de 2015. En colaboración con la Fundación General CSIC, reunió a algunos de los mayores expertos en la materia para analizar cómo reducir este grave problema de salud pública.

1. Pharmacological control
of energy expenditure and obesity
with PI3K inhibitors
in mice and monkeys
Spanish National Cancer Research Center (CNIO)

2. Reducing the IIS pathway:
Akt
PI3K
insulin/IGF1
-
-
Foxo mTORC1
S6K
IRS
Raptor
Lts8
Pten
The insulin/IGF1 signaling (IIS) pathway in healthspan and lifespan
anabolism
CR
RAPA
# increases longevity
• in mice, flies, worms
• in humans (FOXO3 alleles
are linked to longevity)
# improves glucose homeost.
• upon aging or high fat
# increases energy expenditure
• lower nutritional storage

3. EDITORIAL
September 13, 2012
Humans with PTEN germline mutations are obese
Genetic evidence in humans also links PTEN with the control of obesity

4. Rationale
anabolism ins. res.
metab. synd.
longevityobesityIIS PI3Kovernutrition
PI3Ki
• CNIO-PI3Ki (a, d)
• GDC-0941 (a, b, g, d)
• BYL-719 (a)

5. • obese mice after 8 months of HFD
• 13-25% body weight gain
-5 -4 0 1 2 3 4 5 6 7 8 9 11 1210 13
CNIO-PI3Ki 10 mg/kg
15 mg/kgCNIO-PI3Ki
GDC-0941 10 mg/kg
GDC-0941 75 mg/kg
vehicle
-3 -2 -1
70
80
90
100
110
0 2 4 6 8 10
time (days)
bodyweightchange(%)
body weight change
** * * * *
** ** * * * ** ** ***
*** ** * * * *
0
50
100
150
200
glucose(mg/dl)
body weight change
at day 10
60
70
80
90
100
110
bodyweightchange(%)
*
***
*
std.
diet
glycemia at day 13
0
average food intake
g/day/mouse
1
2
3
4
Weight loss after 2 weeks of PI3Ki treatment
Ortega-Molina et al., Cell Metab. 2015

6. Decreased adiposity after 2 weeks of PI3Ki treatment
vehicle
day -7 day 7 day 14
CNIO-
PI3Ki
15mg/kg
adiposity by DXA
14.2%6.5%
CNIO-PI3Ki GDC-0941
0
10
20
30
40
vehicle 10mg/kg 15mg/kg 10mg/kg 75mg/kg
fat(%)
0
10
20
30
40
fat(%)
**
0.06
SD
SD
0
5
10
15
20
25
30
35
lean(g)
0
5
10
15
20
25
30
35
lean(g)
lean mass
CNIO-PI3Ki GDC-0941
vehicle 10mg/kg 15mg/kg 10mg/kg 75mg/kg
D -7
D 7
D 14
fat mass
Ortega-Molina et al., Cell Metab. 2015

7. vehicle
CNIO-PI3Ki 10
mg/kg
GDC-0941
10mg/kg
GDC-0941 75
mg/kg
liver
CNIO-PI3Ki 15
mg/kg
heart/lung
Reduced liver steatosis and pericardial fat after 2 weeks of PI3Ki treatment
Ortega-Molina et al., Cell Metab. 2015

8. Mechanisms of action of PI3Ki
H&E Ucp1
vehicle
PI3Ki
BAT
0
2
4
6
8
10
foldchange
** **
*
* *
Ortega-Molina et al., Cell Metab. 2012
brown
*
- - -
foldchange
forsk: + + +
100
150
200
250
*
***
**
Ucp1
pre-brown
- - -+ + +
PI3Ki
control
20
*
**
** **
pre-brown brown
3T3-L1
Cebpβ
3T3-L1
Cebpβ
40
30
10
0
50
25
0
Pgc1a
**
PI3Ki
control

9. Mechanisms of action of PI3Ki
April 2015
Figure adapted from:

10. Long-term PI3Ki treatment reduces high fat diet-induced obesity
body weight during PI3Ki treatment
(drinking water)
20
30
40
50
60
0 10 20 30 40 50 60 70
time (days)
bodyweight(g)
HFD: vehicle
HFD: CNIO-PI3Ki
SD: vehicle
SD: CNIO-PI3Ki
24%
Ortega-Molina et al., Cell Metab. 2015
# Body weight reduction is not continuous, but reaches an equilibrium
# Body weight reduction is only observed under overnutrition
Note: Dosing is 0.1 mg/ml in the drinking water, producing an average of ~60 ng/ml serum
concentration, which is ~40x lower that the concentration achieved by gavage of 15 mg/kg.
HFD: high fat diet
SD: standard diet

11. Long-term treatment with CNIO-PI3Ki protects from steatosis and reduces lipid stores
F4/80H&E
1
0
x
SD-vehicle SD-CNIO-PI3Ki HFD-vehicle HFD-CNIO-PI3Ki
H&EOilRedO
livereWAT
Ortega-Molina et al., Cell Metab. 2015

12. Mechanisms of action of PI3Ki
Ortega-Molina et al., Cell Metab. 2015

13. Long-term PI3Ki treatment reverts high fat diet-induced obesity
0 20 40 60 140 160 180 200
80
100
120
140
time (days)
bodyweightchange(%)
vehicle
CNIO-PI3Ki
no treatment
no treatment
obese mice under continuous HFD
Ortega-Molina et al., Cell Metab. 2015
# No evidence for the emergence of resistance
# No evidence for irreversible metabolic changes
# No evidence for toxicities

14. Long-term PI3Ki treatment reverts high fat diet-induced obesity
vehicle
CNIO-Pi3Ki
Ob/Ob mice
Ortega-Molina et al., Cell Metab. 2015
# PI3Ki is effective under overnutrition conditions
(even if the diet is “balanced”)

15. Long-term PI3Ki treatment reverts high fat diet-induced obesity
Ortega-Molina et al., Cell Metab. 2015
# Overnutrition (even in the absence of pre-existent obesity) is
sufficient for the activity of PI3Ki
# Lab-to-lab reproducibility

16. Other properties of CNIO-PI3Ki
Ortega-Molina et al., Cell Metab. 2015
• The drug does not cross the blood-brain barrier
• No alterations on the arcuate nucleus of the hypothalamus
(expression of orexigenic and anorexigenic neuropeptides)

17. Preliminary dosing study in Rhesus monkeys at the NIA-NIH (Rafael de Cabo)
Ortega-Molina et al., Cell Metab. 2015
The highest dose tested of 2.1 mg/kg i.v. was considered safe.
Note. the following assay using obese macaques was done with the same dose
but per os and this gave a serum concentration of drug at 2h of ~30 ng/ml.

18. Assay in Rhesus monkeys at the NIA-NIH (Rafael de Cabo)
Ortega-Molina et al., Cell Metab. 2015
Note: Dosing is 2 mg/kg p.o. which produced a peak at 2h of ~30 ng/ml serum concentration.
This is ~80x lower than the peak achieved in mice with 15 mg/kg p.o. or ~2x lower than the continuous
concentration achieved in mice with 0.1 mg/ml in the drinking water.

19. Summary of the data in macaques
Ortega-Molina et al., Cell Metab. 2015
Treatment of obese macaques with low and chronic doses of CNIO-PI3Ki results in:
• Reduction of adiposity
• A tendency to correct fasting glycemia
• No evidence of toxicities, morbidities, or discomfort
The dose used in the macaques is extremely low, in the order of ~80x lower than
the one used in mice under similar administration conditions.
• Mice: 15 mg/kg p.o., which results in a serum peak of drug at 2h of ~2 µg/ml
• Macaques: 2 mg/kg p.o., which results in a serum peak of drug at 2h of ~30 ng/ml
Based on the preliminary escalation assay, we anticipate that the dose could be
increased at least by 10x.

20. Tumor Suppression Group
Assays with monkeys:
Rafael de Cabo, NIA-NIH, Baltimore
NIH Animal Center, Poolesville
Experimental Therapeutics
Joaquin Pastor
Sonia Martinez
Ana Ortega-Molina (now at MSKCC, NY)
Elena Lopez-Guadamillas
Analysis of the hypothalamus:
Miguel Lopez
CIMUS, Santiago de Compostela
Spanish National Cancer Research Centre (CNIO)