Transcript UVA LVG SphK Inhibitors - License a U.Va. Discovery

Sphingosine Kinase 2 Inhibitors for Acute Kidney
Injury (AKI) and Chronic Kidney Disease (CKD)
Non-confidential Presentation
February 2016
Executive Summary

Mechanism of Action: Best-in-class novel small molecule sphingosine kinase
2 (SphK2) inhibitors that improve kidney endothelial function

Initial Indication: Prevention of acute kidney injury after cardiopulmonary
bypass surgery (Phase II trial in high risk diabetic/CKD patient populations)

Other Indications: Contrast-induced nephropathy, diabetic nephropathy,
chronic allograft nephropathy, transplantation associated ischemia

Patent Protection: 2 Patents issued, 3 pending

Route of Administration: i.v. for AKI and oral for CKD

Development Phase: Lead optimization (12 months to IND candidate)

Competition: Very few programs in clinical development, none have
progressed past Phase II; none targeting endothelium directly
2
The Problem: Kidney Disease
Acute Kidney Injury (AKI):
• Affects 1.2MM people/year in US
• Main causes: cardiac surgery, sepsis,
nephrotoxic drugs, contrast-imaging
• No FDA-approved treatments
• 25% mortality; 30% to CKD
• Costs US >$10B/year (hospital stay,
dialysis)
Chronic Kidney Disease (CKD):
Capillaries
• Affects 26MM+ people in US
• Main causes: diabetes/hypertension
• Treated with anti-hypertensive
medications with only modest efficacy
• Renal failure requires dialysis or
transplant
• >25% of Medicare spending (dialysis, 3
transplant)
AKI is a
Significant
Unmet
Medical
Need
Lewington AJP et al. Raising Awareness
of Acute Kidney Injury A Global
Perspective of a Silent Killer. Kidney Int.
2013;84(3):457-467.
4
AKI & CKD as
Interconnected
Syndromes
 Rapid
(AKI) vs slow
(CKD) decline in kidney
function
 AKI is a major risk factor
for CKD and vice versa
 Shared disease biology 5
Chawla LS et al. Acute Kidney Injury and Chronic Kidney Disease as Interconnected Syndromes. N Engl J Med 2014;371:58-66.
Acute Kidney Injury (& CKD) Proof
of Concept- UVA’s Thesis
 Endothelial dysfunction is a critical driver of both AKI & CKD
 The sphingosine 1-phosphate (S1P) gradient maintains endothelial barrier
integrity in humans
 UVA LVG’s SphK2 inhibitors can steepen the S1P gradient and provide
protection to the endothelium during acute and chronic renal injury, preventing
further decline in renal function
 S1P and creatinine are easily measurable biomarkers to track these effects in
patients
 Prevention of AKI (rise in creatinine) in a high-risk diabetic/CKD population
during cardiopulmonary bypass surgery is a reasonable clinical PoC for AKI
and would provide a strong signal for the utility of an SphK2 inhibitor to treat
diabetic nephropathy (CKD) patients
6
Endothelial Dysfunction is a Widely
Acknowledged Driver of AKI &
CKD...
Immune cell
Crosssection of a
Blood
Vessel
Pericy
te
Red blood
cell
Fibrosi
s
Endothelial
cells
Vascular Leak
Inflammation
Blood Flow
Renal Function
7
…Yet There Are No Drugs in Clinical
Development for AKI That Directly Address
Endothelial Dysfunction
Discovery
Phase I
CXA-10
(Nrf2); 10-nitrooleic-acid
AKI COMPETITIVE
LANDSCAPE
HIF
BB3 (HGF)
Phase II
THR-184 (peptide)
(BMP/SMAD pathway)
recAP
(recomb. Alk. Phosp.)
Bendavia™ (Mitochondrial
targeting agent)
Tie2 Receptor Agonist
QPI-1002 (I5NP) (p53 siRNA)
Novartis option
Pyridorin
(scavenger of pathologic
oxidative chemistries)
CMX-2043 (chemically modified
alpha lipoic acid; AKT activation)
Phase III
• Currently, no
phase III
programs
• No clinical
programs
targeting
endothelial
dysfunction
directly (most
directed at
tubular
epitheilum)
• No SphK
inhibitor
programs in
development for
AKI
8
OPN-305 (Anti-TLR Antibody)
The S1P Gradient Serves to Maintain
Proper Endothelial Function via Tonic
Stimulation of Endothelial S1P1
Receptors
Interstitial space
Pericyte
Adherens
Junction
Endothelial cell
N-cadherin
VEcadherin
Rac
S1P1
S1P
Albumin
S1P
ApoM
Blood vessel
Figure 1. The S1P Gradient Maintains The
UVA Researchers
developed
a novel
Integrity of thehave
Endothelium:
High
plasmaway to
levels of the
S1PS1P
tonically
signal through
increase
gradient
and protect the
endothelial S1P1
receptors leading to Rac
endothelium
activation and stabilization of adherens
junctions. Endothelial S1P1 activation also
9
Background: S1P is an Essential
Lipid
S1P:

Maintains blood vessel integrity


Maintenance of endothelial barrier function
Pericyte adhesion

Is necessary for proper lymphocyte
trafficking
 Mobilizes hematopoietic stem cells
10
S1P Signaling – Key Concepts
Plasma sources – RBCs ~70%, endothelium ~30%,
platelets ~1%
 Compartmentalization (S1P Gradient)



Rapid turnover



Synthesis (SphK1/SphK2)
S1P Lyase and Phosphatases (degradation in tissue) or
Sphk2 (removal from blood)
Highly bound to plasma proteins



Blood vs tissue levels
ApoM-containing HDL particles (2/3)
Albumin (1/3)
Five S1P receptors (GPCRs)

S1P1R most prominent
11
The S1P Cycle
S1P1-5R
S1P
Spns2
phosphatase
S1P
S1P Sph + Pi
SphK1,2
de novo
Cer
S1P
Sph
S1Pase
Sph
S1P lyase
hexadecenal + Et-P
Figure 2. S1P Cycle: S1P is synthesized intracellularly
by SphKs and either degraded or released into the
extracellular environment via SPNS2 and other
12
Sphingosine Kinases
SphK1





SphK2
Cytoplasmic localization
Inducible expression (e.g.
growth factors); highest in
lung, spleen and leukocyte
Trans-locates to cell
membrane
KM=14 µM for sphingosine
Implicated in cancer,
inflammation
C
1
C
1
NLS
C
2
C
2
BH3




C
3
C
4
C
3
C
4
Cytoplasmic & nuclear
localization
More static expression
than SphK1; highest in
kidney and liver
KM=3.4 µM for sphingosine
Widely disputed biology,
particularly role in nucleus
(HDAC)
C
5
SphK1
C
5
SphK2
TM
13
Blood S1P Levels in SphK Knockout
Mice
Circulating S1P levels in mice
n= 4 WT, 9 Sphk1‐/- and 6
Sphk2‐/- mice.
The null mice were either
heterozygous or wild type at
the other SphK locus
LC/MS data
***p < 0.001 (one-way
ANOVA, and Bonferroni’s
multiple comparison test,
compared with WT)
Kharel et al, Sphingosine kinase type 2 inhibition elevates circulating sphingosine 1phosphate. Biochem J. 2012 Oct 1;447(1):149-57.
14
Tissue Restriction of S1P Pathway
Enzymes and Their Role in Maintenance
of the S1P Gradient
RBCs
Responsible for
SphK1synthesis of 2/3 of
Plasma S1P
SphK2
S1P Lyase
Endothelium
“Tissue”
Responsible for
synthesis of 1/3 of
Plasma S1P
Responsible for
synthesis of 1/2 of
Tissue S1P
Minimal
Responsible for
clearance of S1P
from Blood
Responsible for
synthesis of 1/2 of
Tissue S1P
None
Responsible for
degradation of S1P
Responsible for
degradation of S1P
15
SphK2 is Involved in the Clearance
of S1P From Blood

Clearance of IV-injected S1P
1.2
1
**
C17-S1P (mM)
RBCs are main source of
plasma S1P and only
utilize SphK1 for
synthesis of S1P
 SphK2 present in tissue
but only minimal in RBCs
 SphK2 appears to play a
key role in clearance of
S1P from blood to tissue
(endothelium likely key
site of action)
0.8
WT
**
0.6
**
WT+434
SphK2-/-
0.4
Note: 434 is an
SphK2-selective
inhibitor
0.2
0
ASAP
5
20
50
Time (min)
Kharel et al J Pharmacol Exp Ther. 2015 Oct;355(1):23-31.
16
How Could a Kinase Be Involved in
Clearance? Analogy to Glucose
Metabolism…

Capturing an imported
metabolite by phosphorylation is
a known mechanism
 Phosphorylation irreversibly
traps the metabolite inside cells
 Classic example: Glucose
Glucose 6-phosphate

http://image.slidesharecdn.com/sach1bi111angl-110919104336phpapp01/95/sach1bi1-11-angl-43-728.jpg?cb=1316429360
Tissue restriction of glucose kinases
drives differential uptake
 Glucokinase (liver) and hexokinase
(skeletal muscle, other tissues)
 Differing enzyme kinetics of
glucokinase and hexokinase also
17
drive differential uptake
Proposed Model For Role of SphK2 in
Clearance of Blood S1P

BLOOD
ENDOTHELIAL
CELL



Sphingosine (Sph) is trapped
irreversibly as S1P
Tissue restriction of SphK2
(i.e. absence of SphK2 in
blood, but presence in
endothelium) limits role to
uptake (as opposed to
synthesis)
SphK2 has a higher affinity
for Sph than SphK1
SphK2 inhibition results in a
rise in blood Sph (in addition
to blood S1P)
18
Effects of SphK2 Inhibition
No effect
Effect on S1P Synthesis:


No effect
Effect on S1P Clearance:

No effect
 S1P in Blood
 S1P in Tissue
(accumulation)
19
Compartmental Pharmacodynamics Vary
With SphK Inhibitor Selectivity
SphK1
Selective
SphK2
Selective
(SphK1 > SphK2)
Equipotent at
SphK1 & 2
200%
(SphK2 > SphK1)
200%
Illustrative
% Change
in Blood 0%
S1P
-100%
% Change
0% in Tissue
S1P
-100%
20
Lead SphK2 Inhibitor – SKX223307
SKX223307
Series
 Low double digit nanomolar potency against SphK2
 Excellent physicochemical and ADME profiles suitable for further
development
 Excellent PK - long-lived in vivo
 Pro-drugs with oral bioavailability have been developed
Competitor SphK Inhibitors
 The majority of, if not all, SphK inhibitors reported are SphK1selective and dual (SphK1 biased)
 ABC294640 (Apogee Biotech) is a low potency SphK2 inhibitor (10
µM at SphK2)1 in Phase I oncology trials. It has been reported to
lower circulating S1P in vivo2
1French
21
KJ et al. Pharmacology and Antitumor Activity of ABC294640, a Selective Inhibitor of Sphingosine Kinase-2. J Pharmacol Exp Ther. 2010 Apr; 333(1): 129–139.
V et al. Antitumor activity of sphingosine kinase 2 inhibitor ABC294640 and sorafenib in hepatocellular carcinoma xenografts. Cancer Biol Ther. 2011 Mar 1;11(5):524-34.
2Beljanski
Pharmacodynamic Response SKX223307 Causes Circulating
S1P to Rise Significantly
Whole Blood S1P
Veh
6
SKX223307
5

S1P uM
4


3
Key
n=3 mice per
arm
5 mg/kg i.p.
Single dose
2
1
Lynch Lab - UVA
0
ASAP
2 hrs
6 hrs
24 hrs
22
SKX223307 Blocks Renal Vascular
Leak in the 3 Day Mouse UUO
Model
Evans Blue ug/g kidney
*
40
 3-day mouse UUO model
 Prophylactic
then
concurrent
treatment
 3 mg/kg i.p. dosing q.d.
 Evans blue dye
 n= 8 vehicle and 9 drug treated
mice
 Statistical Analysis: One-Way
ANOVA with Newman-Keuls
Multiple comparison test.
*p<0.05 and ***p<0.001
***
***
30
Key
20
10
3m
g/
2I
kg
SK
Ve
hi
cl
30
e
U
nl
ig
at
ed
7 Vehicle
U
nl
ig
at
Ve
ed–
3m SKX223307
hi
g/
3clemg/kg
kg
l ig
SK
at
ed
2I
3Vehicle
07
lig
a
SKX223307ted–
3 mg/kg
0
Unligated
Ligated
Ferrer Lab - UCHC
23
Dose-dependent Protection From
AKI in the 26’ Bilateral IRI Model –
SKX223307
p< .001
Key
p= .003
SKX223307 – 2.0 mg/kg – IRI
SKX223307 – 0.2 mg/kg – IRI
SKX223307 – .02 mg/kg – IRI
Vehicle-IRI
Sham
SKX223307 – 2.0 mg/kg – IRI
SKX223307 – 0.2 mg/kg – IRI
SKX223307 – .02 mg/kg – IRI
Vehicle-IRI
Sham
 n=4 mice per
treatment arm
 Creatinine
measurement 24
hours post-reperfusion
 ‘307 given 2 hrs before
surgery, i.p
 Acute Tubular Necrosis
(ATN) Scoring: 5-10 fields
from each of cortex,
medulla, and inner
medulla were evaluated
scored and averaged
 0=normal
 1= <10% ATN
 2= 10-25% ATN
 3= 26-75% ATN
 4= >75% ATN
Okusa Lab - UVA
24
Significant Treatment Effect of
SKX223307 in the 26’ Bilateral IRI
Model
Key
Prophylactic
mode
Therapeutic
Mode
 n=3 mice per treatment
arm
 Creatinine measurement
24 hours post-reperfusion
 Arm 1: 2 mg/kg ‘307 given
2 hrs before surgery, i.p
 Arm 2: 2 mg/kg ‘307 given
1 hr after reperfusion, i.p.
Okusa Lab - UVA
25
SKX223307 Significantly Blocks
Renal Fibrosis in the 7 Day UUO
Rat Model
450
400
350
p<0.05
μg Hydroxyproline
per gram of wet kidney
300450
p<0.001
Key
250400
Unligated
200350
300
150
250
100
200
50
150
0100
50
1
2
3
4
3
SKX111057 5
mg/kg
(SphK1 Inhibitor)
2
SKX223307- 5
mg/kg
1
SKX223307- 2
mg/kg
Vehicle
0
Ligated
 n=4 mice per treatment
arm
Unligated
 7 day model
 Ligated
Treatment initiated day of
surgery (post-surgery) and
continued once a day for 7
days
 Note: treatment with
SphK1 inhibitor ineffective
4
26
Lynch Lab - UVA
Intellectual Property


US 8,686,046 Filed 10/2/10
US 9,421,177 Filed 2/13/12

PCT/US2013/025341Filed 9/30/15



PCT/US2015/053315 Filed 2/8/13



Published WO2013/119946 8/15/13
National Stage US, EP, HK
Published WO2016054261 A1 4/7/16
National Stage Entry due 4/1/17
U.S. Provisional 62/315,193 Filed 3/30/16
27
Summary

AKI and CKD are significant unmet medical needs / market opportunities that share a common
disease biology in endothelial dysfunction

UVA LVG’s SphK2 inhibitors drive a measurable increase in the S1P gradient and enhance the
integrity of the renal endothelium

UVA LVG’s SphK2 inhibitors have clear dose dependent reno-protective effects in both
prophylactic and therapeutic modes in the IRI model of AKI. Protective effects are also present
in the UUO model of renal injury/fibrosis

S1P is an easily accessible, circulating biomarker to measure target engagement and aid in
clinical development of SphK2 inhibitors

Little competition in AKI R&D; highly differentiated mechanism of action

UVA LVG’s SphK2 inhibitors are novel and suitable for further development for AKI and CKD

Clinical PoC would focus on preventing AKI in diabetics in cardiopulmonary bypass surgery
setting; also provides insight into potential as CKD therapy

With the appropriate investment an IND candidate molecule could be delivered in 12 months
Seeking strategic partner for investment, license,
and or research collaboration.
28
APPENDIX
29
Clinical Evidence That Modulation
of the S1P Pathway Can be Used
to Therapeutic Benefit

Gilenya® (fingolimod) – is a functional
antagonist of S1P1R that is approved and
marketed for the treatment of multiple
sclerosis

Drives lymphopenia in patients
 Side effects: 1st dose bradycardia, macular
edema
30
Selected References
Endothelial Dysfunction in AKI

Molitoris BA. Therapeutic translation in acute kidney injury: the epithelial/endothelial axis. J Clin Invest. 2014 Jun;124(6):2355-63.

Verma SK et al. Renal Endothelial Injury and Microvascular Dysfunction in Acute Kidney Injury. Semin Nephrol. 2015 Jan;35(1):96107.
The S1P Gradient and Endothelial Barrier Integrity

Olivera A et al. Shaping the landscape: metabolic regulation of S1P gradients. Biochim Biophys Acta. 2013 Jan;1831(1):193-202.

Sensken SC et al. Redistribution of sphingosine 1-phosphate by sphingosine kinase 2 contributes to lymphopenia. J Immunol. 2010
Apr 15;184(8):4133-42.

Camerer E et al. Sphingosine-1-phosphate in the plasma compartment regulates basal and inflammation-induced vascular leak in
mice. J Clin Invest. 2009 Jul;119(7):1871-9.

Marsolais D et al. Chemical modulators of sphingosine-1-phosphate receptors as barrier-oriented therapeutic molecules. Nat Rev
Drug Discov. 2009 Apr;8(4):297-307.

Itagaki K et al. Sphingosine 1-phosphate has dual functions in the regulation of endothelial cell permeability and Ca2+ metabolism.
J Pharmacol Exp Ther. 2007 Oct;323(1):186-91.
S1P and Pericyte Recruitment/Adhesion

Liu Y et al. Edg-1, the G protein-coupled receptor for sphingosine-1-phosphate, is essential for vascular maturation. J Clin Invest.
2000 Oct;106(8):951-61.
S1P and AKI

Bartels K et al. Sphingosine-1-phosphate receptor signaling during acute kidney injury: the tissue is the issue. Kidney Int. 2014
Apr;85(4):733-5.

Ham A et al. Selective deletion of the endothelial sphingosine-1-phosphate 1 receptor exacerbates kidney ischemia–reperfusion
injury. Kidney Int. 2014; 85: 807–823.

SW Park et al. Sphingosine kinase 1 protects against renal ischemia-reperfusion injury in mice by sphingosine-1-phosphate1
receptor activation. Kidney Int. 2011 Dec;80(12):1315-27.
Effect of SphK Inhibitors on S1P Levels

Kharel et al. Sphingosine kinase type 1 inhibition reveals rapid turnover of circulating sphingosine 1-phosphate. Biochem J. 2011
Dec 15;440(3):345-53.

Kharel et al. Sphingosine kinase type 2 inhibition elevates circulating sphingosine 1-phosphate. Biochem J. 2012 Oct 1;447(1):14957.

Kharel et al. Sphingosine Kinase 2 Inhibition and Blood Sphingosine 1-Phosphate Levels. J Pharmacol Exp Ther. 2015
Oct;355(1):23-31.
31
AKI and CKD as Interconnected Syndromes

Chawla LS et al. Acute Kidney Injury and Chronic Kidney Disease as Interconnected Syndromes. N Engl J Med 2014;371:58-66.