Compounds related to 3-alkylamine-1H-indolyl acrylate and ...
Transcript of Compounds related to 3-alkylamine-1H-indolyl acrylate and ...
XIII Encuentro de Cooperación Farma-Biotech
Barcelona, october 20th 2015
Compounds related to 3-alkylamine-1H-indolyl acrylate and their use for the treatment of neurodegenerative diseases
Rafael León Ph.D.
Content
1. The Institution
2. The Product a) Target Indications b) Innovative mechanisms of action c) Differential features facing the market d) Current status of development e) IPR protection f) Pitfalls & Risks to be considered
3. Partnering Opportunities
XIII Encuentro de Cooperación Farma-Biotech
INSTITUTION: IIS HOSPITAL UNIVERSITARIO LA PRINCESA
ITH 8: Rafael León Ph.D (Medicinal Chemistry and biological screening)
Dr. Leon MedChem group: Current members
IIS H. La Princesa(Fundación I. B.)
Area 1: Francisco Sánchez-Madrid Area 2: Antonio García García Area 3: Isidoro González Álvaro
• Giammarco Tenti: Postdoctoral Student
• Zulay Pardo: Postdoctoral Student (Juan de la Cierva fellowship)
• Patrycja Michalska: Ph.D. Student (FPU fellowship)
• Isabel Gameiro: Ph.D. Student (FPI fellowship)
• Sheila Abril: Ph.D. Student (FPU fellowship)
• Michelle Satriani: Erasmus Student
• Guillermo Furones: Pharmacology Master Student
• Alberto Ruiz Priego: Pharmacology Master Student
Neuropharmacology and neuroprotection
Hippocampal Neurotransmision
Clinical pharmacology and pharmacogenomics Affective disorders Epilepsy
neurosurgery Cardiovascular
diseases
Dr. Leon MedChem group: Financial support
Content
1. The Institution
2. The Product a) Target Indications b) Innovative mechanisms of action c) Differential features facing the market d) Current status of development e) IPR protection f) Pitfalls & Risks to be considered
3. Partnering Opportunities
XIII Encuentro de Cooperación Farma-Biotech
Neurodegenerative diseases: Cognitive degeneration and chronic neuroinflammation
Neurodegenerative and inflammatory diseases
• Common physiopathological hallmarks: Directed drug design
q Aberrant protein aggregates q Mitochondrial dysfunction
q Extensive oxidative stress
q Neuroinflammation q Autophagy failure
q Detoxification pathways failure
q Synaptic deregulation
Target indications
Neurodegenerative and inflammatory diseases
• Neurodegenerative diseases (600 pathologies described as NDDs)
q Alzheimer disease (claim 11) q Parkinson’s disease (claim 12)
q Huntington’s disease (claim 13)
q Multiple sclerosis (claim 14) q Cerebral ictus (claim 15)
q Peripheral neuropathic diseases (claim 16)
q Amyotrophic lateral sclerosis (claim 17)
Potential target indications • Wide range of application not included in the first patent due to its mechanism of action
q Cancer q Chronic obstructive pulmonary disease (seeking financial support, ITH-Evgen pharma, UK)
q Retinal neurodegenerative diseases (In vitro and in vivo studies in progress, UA-ITH-Bayer Healthcare)
q Chronic kidney disease (In vitro and in vivo studies in progress, FJD-ITH) q Crohn disease
Content
1. The Institution
2. The Product a) Target Indications b) Innovative mechanisms of action c) Differential features facing the market d) Current status of development e) IPR protection f) Pitfalls & Risks to be considered
3. Partnering Opportunities
XIII Encuentro de Cooperación Farma-Biotech
Nrf2-ARE pathway… combined with:
Nrf2-ARE pathway as innovative mechanism of action
Oxidative stress regulation and neuroinflammation
Nrf2
HS
HS
HS
SH
SH
SH
Ub
Proteasome Degradation
EpRE
Maf
Gene transcription
A) Normal conditions
Phase II Antioxidant genes- Direct antioxidant enzymes: HO-1- Glutation homeostasis: GCLc, GCLm- Detoxifiying enzymes: GST, NQO1- Thiol homeostasis: Txn, TrxR1, Srx1- Inhibition of inflammation: HO-1
Keap1dimer
Proteasome 26S
Nrf2Ub
Cul3
BTB BTB
Nrf2
HS
HS
HS
SH
SH
SH
Ub
CuI3
BTB BTB
B) Oxidative conditions
Nrf2
S
HS
HS
SH
SH
BTB BTB
Nrf2
HS
HS
HS
SH
SH
SH
BTB BTB
C) Receptor modulation
STRESORSElectrophilesROSHeavy metals
Direct Nrf2-Keap1 inhibitors
R
Hinge and latch mechanism
D) Chronic inflammation
AKT
P38
GSK3β
HDAC
Fyn Nrf2P
CuI3
nAChR-α7 MT 1/2 recptors
PI3K
Jak2
GSK3β
Nrf2
Nrf2
Nrf2
ERK
Prolonged inflammation
León et al, Pharm. & Therap. 2015, in press.
• High CNS vulnerability • Crosslinks between
oxidative stress and protein aggregates
• Mitochondrial dysfunction
Neuronal loss
Sultana, R.; Butterfield, D. A. J Alzheimers Dis 2010, 19, 341-53.
Glass et al, Cell 2010, 140, 918-34.
• Oxidative stress • Neuroinflammation
• Glial overactivation (ROS and protein agregates)
• Pro-inflammatory cytokines and infiltration
Nrf2-ARE pathway target of BG12 (Tecfidera®)
Nrf2-ARE pathway as innovative mechanism of action
Sultana, R.; Butterfield, D. A. J Alzheimers Dis 2010, 19, 341-53.
Glass et al, Cell 2010, 140, 918-34.
2nd generation of Nrf2 inducers are generating great interest
Nrf2-ARE pathway as innovative mechanism of action
Sultana, R.; Butterfield, D. A. J Alzheimers Dis 2010, 19, 341-53.
Glass et al, Cell 2010, 140, 918-34.
q XenoPort: Biopharmaceutical company is developing a “ME TOO” derivative of Tecfidera
Content
1. The Institution
2. The Product a) Target Indications b) Innovative mechanisms of action c) Differential features facing the market d) Current status of development e) IPR protection f) Pitfalls & Risks to be considered
3. Partnering Opportunities
XIII Encuentro de Cooperación Farma-Biotech
3-alkyl-1H-indolyl acrylate derivatives Vs BG12 (Tecfidera®)
Advantages and differences of NCE facing the market
• Bardoxolone RTA-402
• Clinical Trial phase III BEACON NCT00664027
• Result: TERMINATED (Safety concerns October 2012)
q Nrf2 induction potency control: 2nd Strong Nrf2 induction is TOXIC¡¡¡
q Nrf2 induction is not enough: 1st Combination of mechanisms (Nrf2 induction – free radical scavenger – anti-inflammatory)
Advantages and differences of NCE facing the market
q 3rd Improved activity without increasing toxicity might led to: q Lower incidence/ less sever side effects
q Improved compliance, fewer treatment failures
q Onset and/or magnitude of immunomodulation q Earlier onset of immunomodulation
Sultana, R.; Butterfield, D. A. J Alzheimers Dis 2010, 19, 341-53.
Glass et al, Cell 2010, 140, 918-34.
• High CNS vulnerability • Crosslinks between
oxidative stress and protein aggregates
• Mitochondrial dysfunction
q 4th Improvement in efficacy due to combination of action mechanism: q Dosing frequency
q Once-a-day rather than BID (TECFIDERA)
q 5th Novel chemical entity with many substitution possibilities: q Pharmacokinetic and pharmacodynamics properties modulation
Content
1. The Institution
2. The Product a) Target Indications b) Innovative mechanisms of action c) Differential features facing the market d) Current status of development e) IPR protection f) Pitfalls & Risks to be considered
3. Partnering Opportunities
XIII Encuentro de Cooperación Farma-Biotech
Development status 3-alkylamine-1H-indolyl acrylate derivatives
In vitro characterization: Nrf2 induction
R = crotonic
Basal 0,3 3 10 300
10
20
30
40
1 (µM)
***
**
***
Luci
fera
se A
ctiv
ity(F
old
incr
ease
a.u
.) R = p-Cl
Basal 0,3 3 10 300
5
10
15
20
10 (µM)
***
***
Luci
fera
se A
ctiv
ity(F
old
incr
ease
a.u
.) R = m-OMe, p-OH
Basal 0,3 3 10 300
1
2
3
4
7 (µM)
***
***
*
Luci
fera
se A
ctiv
ity(F
old
incr
ease
a.u
.)
Basal 0.3 3 10 300.0
0.5
1.0
1.5
Melatonin (µM)
***
Luci
fera
se A
ctiv
ity(F
old
incr
ease
a.u
.)
Basal 0.3 3 10 300.0
0.5
1.0
1.5
Ethyl cinnamate (µM)
*****
Luci
fera
se A
ctiv
ity(F
old
incr
ease
a.u
.)
Basal 0.3 3 10 300.0
0.5
1.0
1.5
2.0
Et. Cin. + Mel (µM)
***
Activ
idad
Luc
ifera
sa(ta
nto
por 1
)
R = crotonic
Basal 0,3 3 10 300
10
20
30
40
1 (µM)
***
**
***
Luci
fera
se A
ctiv
ity(F
old
incr
ease
a.u
.) R = p-Cl
Basal 0,3 3 10 300
5
10
15
20
10 (µM)
***
***
Luci
fera
se A
ctiv
ity(F
old
incr
ease
a.u
.) R = m-OMe, p-OH
Basal 0,3 3 10 300
1
2
3
4
7 (µM)
***
***
*
Luci
fera
se A
ctiv
ity(F
old
incr
ease
a.u
.)
Basal 0.3 3 10 300.0
0.5
1.0
1.5
Melatonin (µM)
***
Luci
fera
se A
ctiv
ity(F
old
incr
ease
a.u
.)
Basal 0.3 3 10 300.0
0.5
1.0
1.5
Ethyl cinnamate (µM)
*****
Luci
fera
se A
ctiv
ity(F
old
incr
ease
a.u
.)
Basal 0.3 3 10 300.0
0.5
1.0
1.5
2.0
Et. Cin. + Mel (µM)
***
Activ
idad
Luc
ifera
sa(ta
nto
por 1
)
Nrf2-induction properties are easily tuneable
Development status 3-alkylamine-1H-indolyl acrylate derivatives
In vitro characterization: Antioxidant effect
Mel
Et. Cina
Mix 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 160
2
4
6
8
10
Antio
xida
nt a
ctiv
ity(T
rolo
x eq
.)
In vitro characterization: Anti-inflammatory effect (LPS response blockade)
Basal LPS 30 60 10075
100
125
150
175
Mn 3/24 ( µM) Pm 15 (µM) Pm 19 (µM)
###
****** ***
1 (µM)
R = Crotonico
LPS (1 µg/mL)
% N
itrite
pro
duct
ion
(% V
s Ba
sal)
Basal LPS 30 60 10075
100
125
150
175
Mn 3/24 (µM)
###
******
12 (µM)
R = m,p-di-Cl
LPS (1 µg/mL)
***
% N
itrite
pro
duct
ion
(% V
s Ba
sal)
Basal LPS 30 60 10075
100
125
150
175
Mn 3/24 ( µM) Pm 15 (µM) Pm 19 (µM)
###
*** ******
7 (µM)
R = m-OMe; p-OH
LPS (1 µg/mL)%
Nitr
ite p
rodu
ctio
n(%
Vs
Basa
l)
Basal 3 10 30
100
150
200
Mel (µM)
****
###
LPS (1 µg/ml)
% N
itrite
pro
duct
ion
(% V
s Ba
sal)
Basal 3 10 30
100
150
200
LPS (1 µg/ml)
Etil Cin (µM)
*###
% N
itrite
pro
duct
ion
(% V
s Ba
sal)
Basal 10 30 100
100
150
200
MIX (µM)
###*
***
**
LPS (1 µg/ml)
% N
itrite
pro
duct
ion
(% V
s Ba
sal)
Basal LPS 3 10 30
100
120
140
160
Mn 3/24 ( µM) Pm 15 ( µM) Pm 19 ( µM)
LPS (1 ng/mL)
###
7 (µM)
***
***
***
R = m-OMe; p-OH
% N
itrite
pro
duct
ion
(% V
s Ba
sal)
Development status 3-alkylamine-1H-indolyl acrylate derivatives
• Bardoxolone RTA-402
• Clinical Trial phase III BEACON NCT00664027
• Result: TERMINATED (Safety concerns october 2012)
Sultana, R.; Butterfield, D. A. J Alzheimers Dis 2010, 19, 341-53.
Glass et al, Cell 2010, 140, 918-34.
• High CNS vulnerability • Crosslinks between
oxidative stress and protein aggregates
• Mitochondrial dysfunction
In vitro characterization: Neuroprotective effect against oxidative stress
Control 0.3 1 3 350
75
100
OGD (15 Min) + 2 h reox
###
***
*** ****
(µM)
Compound 7Melatonin
% C
ell V
iabi
lity
(MTT
Red
uctio
n)
q Cerebral ictus (hippocampal slices) q Tau hyperphosphorylation
Neurodegenerative diseases specific models
Control 0.3 1 3 3
50
75
100
Okadaic acid (30 nM)
*** ******
###
(µM)
Compound 7Melatonin
% C
ell V
iabi
lity
(MTT
Red
uctio
n)
BASAL Mel 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 160
20
40
60
80
100
R/O (30/10 µM) + Compound (1 µM)Pre-incubation (1 µM)
R/O 30/10 µM
###
******
*** *** *** *** *** *** *** *** ******
******
******
***
% C
ell V
iabi
lity
(MTT
Red
uctio
n)
BASAL Mel 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 160
20
40
60
80
100
Menadione (8 µM) + Compound (1 µM)Pre-incubation (1 µM)
Menadione 8 µM
###
***
***
*** *** *** ***
***
***
***
***
***
***
***
*** ****** ***
% C
ell V
iabi
lity
(MTT
Red
uctio
n)
Development status 3-alkylamine-1H-indolyl acrylate derivatives
Sultana, R.; Butterfield, D. A. J Alzheimers Dis 2010, 19, 341-53.
Glass et al, Cell 2010, 140, 918-34.
q Amyloid β and Okadaic acid combination
Basal 1 10 300
20
40
60
80
100
Mn 3/24 (µM) Pm 15 (µM) Pm 19 (µM)
Aβ (0.5 µM) /Okadaic acid (10 nM)
###
Melatonin
% C
ell V
iabi
lity
(MTT
Red
uctio
n)
RO
S Ib
a-1
Basal 1 10 300
20
40
60
80
100
***
###
**
***
Mn 3/24 (µM)
Aβ (0.5 µM) /Okadaic acid (10 nM)Compound 1 (µM)
% C
ell V
iabi
lity
(MTT
Red
uctio
n)
Neuroinflammation inhibition (Primary rat glial cultures)
q Nitrite release reduction
Basal 3 10 30
100
120
140
160
Mn 3/24 (µM) Pm 15 (µM) Pm 19 (µM)
LPS (1 µg/mL)
###
Compound 7
***
***
***
% N
itrite
pro
duct
ion
(% V
s Ba
sal)
q TNFα release inhibition
q Lipopolysaccharide model of inflammation
Basal LPS 3 10 300
100
200
300
400
Mn 3/24 (µM) Pm 15 (µM)
LPS (1 µg/mL)
###
7 (µM)
***
% T
NFα
pro
duct
ion
(pic
ogra
ms/
ml)
***
Development status 3-alkylamine-1H-indolyl acrylate derivatives
Sultana, R.; Butterfield, D. A. J Alzheimers Dis 2010, 19, 341-53.
Glass et al, Cell 2010, 140, 918-34.
Compound 7 Vs BG-12
Wilms, H.; J. Neuroinflammation 2010, 7, 30.
q Nrf2 induction
LPS 3 10 300
20
40
60
80
100
Mn 3/24 (µM)
LPS (1 µg/mL)
###
Compound 7 (µM)
***
***
***
**
% T
NFα
pro
duct
ion
(pic
ogra
ms/
ml)
Treatment with 10 ngr/mL LPS and 10 µM BG-12
q TNFα release inhibition q TNFα release inhibition
q Nitrite release reduction q Nitrite release reduction
Compound 7 BG-12
LPS 3 10 300
20
40
60
80
100
Mn 3/24 (µM)
LPS (1 µg/mL)
###
Compound 7 (µM)
***
***
***
% N
itrite
pro
duct
ion
(Nor
mal
ized
to L
PS)
Development status 3-alkylamine-1H-indolyl acrylate derivatives
Sultana, R.; Butterfield, D. A. J Alzheimers Dis 2010, 19, 341-53.
Glass et al, Cell 2010, 140, 918-34.
Differentiating aspects facing commercialization:
100
120
140Nrf2 +/+Nrf2 -/-
LPS (1 µg/ml)7 (10 µM)
ns
ns
###
###
******
Basal% N
itrite
pro
duct
ion
(% V
s B
asal
)
q Nitrite release reduction
Anti-inflammatory effect in Nrf2 KO mouse (Primary mouse glial cultures)
q iNOS expression inhibition
0
1000
2000
3000
4000Nrf2 +/+Nrf2 -/-
LPS (1 µg/ml)7 (10 µM)
ns
ns
###
###
******
Basal
iNO
S/β
-act
in
q HO-1 expression
0
500
1000
1500 Nrf2 +/+Nrf2 -/-
LPS (1 µg/ml)7 (10 µM)
###
***
Basal
HO
-1/β
-act
in
THE ANTI-INFLAMATORY EFFECT IS NOT Nrf2-DEPENDENT
Content
1. The Institution
2. The Product a) Target Indications b) Innovative mechanisms of action c) Differential features facing the market d) Current status of development e) IPR protection f) Pitfalls & Risks to be considered
3. Partnering Opportunities
XIII Encuentro de Cooperación Farma-Biotech
IPR status
3-alkylamine-1H-indolyl acrylate derivatives
• Patent status: Derivatives of 3-alkylamine-1H-indolyl acrylate and their use for the treatment of neurodegenerative diseases
q 19 claims
q Chemical core structure protected
q Chemical substructure protected
q Biological activity protected
q Chemical substituents included
q Anti-inflammatory effect protected
q Use as drugs for Neurodegenerative diseases included as claim
q Common formulations, salts and excipients included for core structure
q Use in COPD, kidney disease or macular degeneration suitable for new patent applications
• Priority date: 15/10/2014 • Application number: P201400810 • Priority country: Spain • PCT application: 15/10/2015 • Priority country: Europe
• Holder: IS Hospital La Princesa / • Universidad Autonoma de Madrid • DNS Neuroscience
• Authors: Rafael León, I. Buendia, E. Navarro, P. Michalska, I. Gameiro, A. López, J. Egea, A. García, M. García.
Content
1. The Institution
2. The Product a) Target Indications b) Innovative mechanisms of action c) Differential features facing the market d) Current status of development e) IPR protection f) Pitfalls & Risks to be considered
3. Partnering Opportunities
XIII Encuentro de Cooperación Farma-Biotech
Pitfalls & risk to be considered
• Bardoxolone RTA-402
• Clinical Trial phase III BEACON NCT00664027
• Result: TERMINATED (Safety concerns october 2012)
Sultana, R.; Butterfield, D. A. J Alzheimers Dis 2010, 19, 341-53.
Glass et al, Cell 2010, 140, 918-34.
• High CNS vulnerability • Crosslinks between
oxidative stress and protein aggregates
• Mitochondrial dysfunction
q Target validation already demonstrated
q Rational design of proposed drug
q In vitro activity demonstrated
q Preliminary toxicity performed: Non-hepatotoxic
Content
1. The Institution
2. The Product a) Target Indications b) Innovative mechanisms of action c) Differential features facing the market d) Current status of development e) IPR protection f) Pitfalls & Risks to be considered
3. Partnering Opportunities
XIII Encuentro de Cooperación Farma-Biotech
Pre-clinical evaluation
Partnering Opportunities
• MS mouse model (MOG injection): Scheduled to be started in January 2016 (Miguel P. Soares Ph.D, European expert in inflammation)
• Pre-clinical toxicity: Big Pharma or CRO companies (Financial support applications: European commission projects)
• ADMET properties • Safety pharmacology –CVS, CNS, RS • Pharmacodynamics • Pharmacokinetics • Acute toxicity: 2 species by 2 routes of administration • Repeat dose toxicity: rodent and non-rodent; two 14 day studies before human trial • Carcinogenicity • Reproductive toxicity: Embryo/foetal development studies – 2 species • Genotoxicity • Mutagenesis: Chromosomal abnormality
Pre-clinical evaluation financial support: Public-private partnership • Innovative Medicines Initiative (IMI): Next call 12 January 2016 (Budget 93M €)
• 3er Health Program: 3 different calls to be open, including preclinical studies projects
• ERA-NET actions under H2020 program: Public-Private partnerships and cofounded actions
• Joint Technology Initiatives: Specific program in H2020
XIII Encuentro de Cooperación Farma-Biotech
Barcelona, 20 de octubre de 2015
Compounds related to 3-alkylamine-1H-indolyl acrylate and their use for the treatment of neurodegenerative diseases
Thank you
!