CYP450 Induction in vitro
Tested in human hepatocytes.
Low potential for drug-drug interactions.
Dimenthyl Sulfoxide
CMX157
CMX157
CMX157
Omeprazole
Phenobarbital
Rifampin
1.00 ± 0.19
1.03 ± 0.24
1.05 ± 0.16
1.08 ± 0.20
44.9 ± 24.5
2.60 ± 0.59
2.04 ± 0.27
0.1% (v/v)
1 M
10 M
100 M
100 M
750 M
10 M
Phenacetin O-dealkylation
(CYP1A2)Treatment Concentration
Bupropionhydroxylation
(CYP2B6)
Testosterone 6 -hydroxylation
(CYP3A4/5)
1.00 ± 0.26
0.954 ± 0.117
0.974 ± 0.098
0.849 ± 0.356
12.8 ± 9.0
20.1 ± 7.8
10.3 ± 4.5
1.00 ± 0.09
1.15 ± 0.09
1.10 ± 0.09
1.40 ± 0.09
2.56 ± 0.42
7.02 ± 0.66
6.32 ± 0.33
Fold Increase
• CMX157 is a novel, lipid conjugate prodrug of tenofovir. It is highly liver targeted, with rapid uptake and e�cient conversion to TFV-PP, the active moiety. CMX157 demonstrated potent anti-HBV activity and was at least as active as TAF in vitro.
• CMX157 showed no signi�cant potential for cytotoxicity, mitochondrial toxicity, CYP450 or drug transporter interactions.
• These results support ongoing clinical studies to determine the safety, pharmacokinetics and antiviral activity of CMX157 in healthy subjects and subjects with chronic HBV infection.
Conclusions
P H A R M A C E U T I C A L S
Preclinical Characterization of CMX157,A Novel Tenofovir Prodrug for the Treatment
of Chronic Hepatitis B InfectionContraVir Pharmaceuticals, Inc. 399 Thornall Street, 1st Floor Edison, NJ 08837
Background
Novel lipid conjugated prodrug of tenofovir designed to utilize lipid uptake pathways in order to:• Increase bioavailability• Enhance target tissue penetration for increased antiviral potency• Decrease renal and bone toxicity by reducing circulating TFV
Method
Results
AD-38 cells: Three independent experiments, run on di�erent days,in triplicate.
CMX157 is at least as active as TAF.
1x10-11
100
90
80
70
60
50
40
30
20
10
0
1x10-10 1x10-9
HBV DNA Inhibition
1x10-8
Inhibitor Concentration (M)
Perc
ent I
nhib
ition
1x10-7 1x10-6 1x10-5
CMX-157
TAF
TDF
Test Article EC50 (nM)Mean ± SD
EC90 (nM)Mean ± SD
CMX-157
TDF
TAF
9.3 ± 3.6
52.7 ± 16.8
32.4 ± 17.1
186 ± 53
776 ± 427
474 ± 261
Antiviral Activity in vitro
AD-38 cells, two separate experiments, triplicate wells.
CMX157 delays viral rebound longer than TAF and other antivirals.Results are consistent across two di�erent experimental conditions.
Viral Rebound in vitro
CMX IC90
110.0%
90.0%
70.0%
50.0%
30.0%
10.0%
-10.0%TAF IC90
HBV
DN
A C
opy
Num
ber
TFV IC90
All Antivirals Tested at IC90
TDF IC90 DMSO 3TC
Day 6
Day 11
Day 15
DMSO
1E+7
1E+6
1E+5
1E+4
1E+3
1E+2
1E+1
1E+0
TDFTFV
TAF
CMX-157 Tet3TC
HBV
DN
A C
opy
Num
ber
Day 6
Day 9
Day 13
All Antivirals Tested at 5 nM
Metabolism in PrimaryHuman Hepatocytes
Rapid, e�cient conversion to TFV-PP.Hepatocytes fully viable.Conversion is time and concentration dependent.
Metabolite Production over Time
0
0.0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1.0
20 40 60 80
Time (hours)
TFV_10 uM CMX-157
TFV_50 uM CMX-157
TFV-PP_10 uM CMX-157
TFV-PP_50 uM CMX-157
Time Point
TFV, nM
0 00
65.191.3238275395
00
70.784.3135143188
46.050.394.9152392460860
00
76101185175361
148
244872
TFV-PP, nM TFV-PP, nMTFV, nM
Systemic, Portal Vein and HepaticPharmacokinetics in Rats
CMX157 20 mg/kg, single oral dose in Sprague-Dawley rats.
Rapid, e�cient extraction and conversion to high liver levels of the active antiviral, TFV-PP after single dose in rat • 86% �rst pass extraction by the liver • Low systemic levels of TFV
CMX157
Drug Concentration in Liver (± SD)
100,000
10,000
1,000
100
100 6 12 18 24
Hours Post Dosing
Conc
entr
atio
n [n
g/g]
TFV
TFV-PP
CMX157 - Systemic Plasma
CMX157 - Portal Plasma
TFV - Portal Plasma
TFV - Systemic Plasma
0
10,000
1,000
100
106 12 18 24
Conc
entr
atio
n [n
g/m
L]
Hours Post Dosing
Drug Concentration in Hepatic Portal Veinvs. Systemic Circulation (± SD)
Whole Body Tissue Distribution in Rats
Single dose, whole body autoradiography in L-E rats.
Highly liver targeted.No substantial accumulation or retention in the heart.
Time after Dose Administration (h) Time after Dose Administration (h)
Tissue Concentrations of Radioactivity After OralAdministration of 20 mg/kg 14C-CMX157 to Rats
Tissue Concentrations of Radioactivity After IVAdministration of 2 mg/kg 14C-CMX157 to Rats
00.010
0.100
1.000
10.000
100.000
1000.000
10 20 30 40 50 00.010
0.100
1.000
10.000
100.000
10 20 30 40 50
Small intestine
Liver
Kidney (cortex)
Kidney (medulla)
Plasma
Heart
Tested in standard tissue culture and vesicle-based assays.
Low potential fordrug-drug interactions.
Compound00
< 10
00
11
< 10< 10
< 100000
0 0
93*32
< 10< 1011
< 10< 10
BSEPMRP2BCRP
OATP1B1OATP1B3
OAT1OAT3OCT2MATE1MATE2-K
P-gp* > 26,000 fold above protein-adjusted Cmax at 400 mg SD
% Inhibition
Cellular Transporter Inhibition in vitro
Tested in human liver microsomes.
Low potential for drug-drug interactions.
CYP450 Inhibition in vitro
Enzyme Zero-Minute Preincubation 30-Minute Preincubation
CYP1A2CYP2B6CYP2C8CYP2C9CYP2C19CYP2D9CYP3A4/5CYP3A4/5CYP3A4/5
3918
4.4*113127122418
3215
3.3*152826121916
Direct Inhibition Time-DependentInhibition
*Value for CYP2C8 is > 3,500 fold above the protein-adjusted Cmax at 400 mg SD
HepG2 cells in a standard assay.
Low potential for mitochondrial toxicity.
Mitochondrial Toxicity in vitro
Viable Cells (% vehicle)
MediumVehicleChloramphenicol
Glucose Galactose Glucose Galactose
CMX 157
TDF
TFV
0.11
10100
0.11
10100
0.11
10100
30
SDH-A: COX-I Ratio
117.3100.0
20.7
83.288.370.4
76.069.362.054.7
* Insu�cient cells for analysis
68.261.757.059.8
2.12.32.02.3
1.71.82.01.8
106.583.265.4
1.92.22.2
2.12.01.9
****
********
72.261.5
94.429.0
2.42.3
1.92.1
14.0 10.1 6.8
102.8100.0
2.32.0
2.21.9
Tested in a large panel of proliferating or non-proliferating cell lines for up to 14 days. Tissues represented include liver, brain, heart, kidney, lymph, muscle, cervix, blood, bone marrow and gut.
Low potential for cytotoxicity.
Cell Line Tissue Origin
3 D
ay In
cuba
tion
6 D
ay In
cuba
tion
12 o
r 13
Day
Incu
bati
on14
Day
Incu
bati
on
HepG2 LiverKidneyBrain
ColorectalPBMC
Muscle
LymphCervicalCervical
LiverLiver
Blood
Blood
Blood
Brain
Liver
Blood
Heart
>100.0>100.0>100.0
>100.0>100.0>100.0
40.216.046.6
61.6
>10.0>10.0>10.0>10.0
8.8>10.0
7.4
7.4
>10.0
>10.0
>10.0
12.5>100.0>100.0>100.0>100.0>100.0
>100.0>100.0>100.0>100.0>100.0>100.0
>100.0
>100.0
>100.0
>100.0
>100.0
28.086.057.7
Proliferating>10.0>10.0>10.0>10.0>10.0>10.0
Non-proliferating
Proliferating
Non-proliferating
>10.0
>10.0
>10.0
>10.0
>10.0
Non-proliferating>100.0 >100.0
Caki-1SNB-78COLO-205CCRF-CEMSJCRH30
CEM-SSHeLaME 180Huh-7HepG2PBMC (stimulated)
PBMC (unstimulated)MonocyteMacrophageDendritic
Hepatocyte
PBMC (unstimulated)
Colorectal 12.9>100.036.0COLO-205Muscle 9.1>100.017.4SJCRH30
Heart 12.8>100.0>100.0Cardiomyocyte
BoneMarrow
1.894.722.46GM-CFU0.53.484.62E-BFU
Cardiomyocyte
Proliferating
Cytotoxicity in vitro
R Rush1, J Greytok2, T Matkovits2, JZ Sullivan-Bólyai2, and D Standring3 1Allon Preclinical Consulting LLC, USA; 2ContraVir Pharmaceuticals Inc., USA;
and 3David Standring Consulting, USA.
Recommended
