AA139 - Adenium Biotech

Report
Corporate presentation,October 2013
Confidential
Ideal Target Product Profile for
multidrug resistant broad spectrum
Gram-negative antimicrobial
•
•
•
•
•
•
Novel mode of action
Bactericidal
Selective and specific
Low frequency of resistance
Active against GAIN pathogens
Drugable
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Arenicin selection process
> 500 organisms screened for
antimicrobial activity
Several G+ but
only one Gidentified !
Clin Cand –
AA139
~40 AMP’s identified
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Mode of action summary
• Arenicin acts at least partly due to non-lipid Amediated penetration and disruption of both Gram
negative membranes
• Inhibition of cytosolic processes in protein synthesis
suggested in TraDIS studies
• Arenicins mode of action is different from Colistins
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Arenicin interacts with the lipid transporter
protein MLAC
•
MlaC is a periplasmic binding protein
maintaining phospholipid homeostasis of
the dual cell membrane
•
Gene analysis of E.coli shows that an
MlaC L11R mutation is required to
prevent the interaction between Arenicin
and MlaC
•
Resistant strains regain sensitiviy to
Arenicin as mutant is not stable
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Arenicin causes loss of cell surface structure
and partial cytoplasm clearing in E. coli
Transmission Electron Microscopy (TEM) of the Arenicin effect on E.coli (ATCC 25922)
E.coli. No treatment. Black arrow,
cytoplasmic membrane; Red arrow, outer
membrane; Green arrow, pili. Bar, 200 nm
E. coli incubated with 32 μg/mL NZ17000
for 40 min induced loss of cell surface
structures and partial clearing of
cytoplasm Bar, 200 nm
University of Queensland 2013
Confidential
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Arenicin causes ATP release without dramatic
changes in cell morphology
Extracellular ATP after 10 min
Fold change
25
20
Ar
15
col
10
pip
5
0
0
At OD600 =0.4 E.coli cells were exposed
to 32ug/ml Arenicin, 64ug/ml Fosfomycin
and 16ug/ml Polymycin B. Even at very
high concentration of Arenicin-3, no
dramatic morphological changes of the
cells were observed.
16
64
256
x MIC
1024 4096
Arenicin-3 (Ar), colistin (col), and
piperacillin (pip) induced release of ATP
from E. coli. Exponential cells were
incubated with drug for 10 minutes and
ATP measured. y-axis is fold change
relative to untreated (0xmic) and x-axis is
fold MIC applied.
Novozymes A/S, 2010
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Arenicin and Colistin have different effects on
osmotic shock genes
Symbol
Gene Colistin Arenicin µg/ml
ID
µg/ml
2½
2½
5
10
Description
Osmotic shock genes
osmB
B1283
osmC
B1482
osmY
B4376
osmE
B1739
bdm
B1481
63
10
5
3
63
29
11
8
6
3
3
lipoprotein, osmotically inducible,
Peroxiredoxin
Osmotically-inducible protein Y precursor
Osmotically-inducible lipoprotein E precursor
Biofilm-dependent modulation proteinosmotically inducible
sra
rcsA
11
6
4
2
3
-
-
rpsV, osmotically inducible
positive regulator for ctr capsule biosynthesis,
B1480
B1951
Whole Genome TraDIS preliminary data suggest inhibition
of key cytosolic processes in gene translation
Novozymes A/S, 2010
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In vitro efficacy summary
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•
•
•
•
•
•
•
•
Potent in vitro activity against GAIN pathogens
Rapidly bactericidal – MBCs ~ MICs
Extremely low spontaneous mutational frequency
Small and mostly reversible increase of MIC in serial
passage studies comparable with Colistin
Little inoculum effect
Moderate effect of serum on MIC
Limited effect of Survanta on MIC
No synergistic or antagonistic effect with other antibiotics
No cross resistance with strains with acquired resistance
to Colistin
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Potent in vitro activity against GAIN
pathogens
MIC90 determinations (MDR clinical isolates)
# strains
AA139
Colistin
Meropenem Ceftazidime Ciprofloxacin Gentamicin Tigecycline
N=325
MIC (µg/ml)
E.coli
N=55
1
0.25
4
>32
>4
>32
0.5
K.pneumonia
N=75
4
8
>16
>32
>4
>32
4
P.aeruginosa
N=75
8
2
>16
>32
>4
>32
ND
A.baumanii
N=120
2
8
>16
>32
>4
>32
4
Eurofins medinet 2012
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Extremely low spontaneous mutational frequency
to Arenicin
Resistance
Frequency
(4XMIC)
Resistance
Frequency
(8XMIC)
Organism
Isolate ID
AA139
AA139
E. coli
ATCC 25922
≤2,50E-12
≤2,50E-12
E. coli
3083559
≤8,90E-11
≤8,90E-11
K. pneumoniae
3083832
≤4,16E-10
≤4,16E-10
K. pneumoniae
3083583
≤1,38E-11
≤1,38E-11
P. aeruginosa
ATCC 27853
≤2,61E-12
≤2,61E-12
P. aeruginosa
3083655
≤2,68E-12
≤2,68E-12
A. baumannii
3083835
≤2,65E-12
≤2,65E-12
A. baumannii
3083684
≤4,80E-10
≤4,80E-10
Eurofins medinet 2012
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Limited effect of mucin (Survanta) on
in vitro efficacy
NBS plates- MHB only
GN_01
E.coli
ATCC 25922
GN_03
K. pneumoniae
ATCC 700603
GN_04
K. pneumoniae
ATCC 13883
GN_34
A.baumannii
ATCC 19606
GN_42
P.aeruginosa
ATCC 27853
GN_43
P.aeruginosa
Polymixin R
GN_44
E.coli
ATCC 10536
GN_45
K. pneumoniae
BAA_2146
Compound ID
MIC [μg/mL]
Colistin
MCC_000094B
Ciprofloxacin MCC_0
00166
AA139
≤0.03
≤0.03
≤0.03
0.03/0.06
1/2
32/64
0.03/0.125
0.06
≤0.03
0.25
≤0.03
0.5/1
2
0.25/0.5
32/64
>64
0.06
0.5
0.125/0.25
0.125
0.25/0.5
2
0.125
1/2
NBS plates- MHB + 5% Survanta
Compound ID
Colistin
MCC_000094B
Ciprofloxacin MCC_0
00166
AA139
MIC [μg/mL]
0.06
1/0.5
0.125
0.5/0.25
0.5
64
0.5/0.25
0.125
≤0.03
2
0.5
>64
1
1
2/1
>64
0.125/0.06
0.125
0.5/0.25
2
0.5
16/8
0.5/0.25
2
8x MIC increase
Decrease in MIC
University of Queensland 2013
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Pharmacokinetic/pharmacodynamic summary
•
•
•
•
Arenicin efficacy is driven by Cmax
Clinical therapy should thus be based on slow bolus injection
Hepatic clearance does not seem to play a role
AA139 has a good volume of distribution corresponding to
the extracellular volume
• AA139 has a half life of 4.3 hours
• AA139 has a low penetration into ELF (<5%)
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PK/PD dose fractionation study shows that Cmax
drives the efficacy of Arenicin (NZ17230)
(5mg/kg over 3 days seems to exert maximal effect)
Tissue burdens following infection with E. coli UT189
1.00E+07
1.00E+06
1.00E+05
CFU/mL homogenate
1.00E+04
1.00E+03
Kidney
1.00E+02
Bladder
1.00E+01
Urine
1.00E+00
1.00E-01
PreTreatment
Q36h
Q18h
Q12h
Q8h
NZ17230 3mg/kg
NZ17230 3mg/kg
NZ17230 3mg/kg
NZ17230 3mg/kg
Euprotec 2013
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In vivo efficacy summary
• Excellent efficacy against K.pneumoniae and E.coli in
UTI with ED50 of 0.5-1mg/kg (BID I.V. administration)
• Modest efficacy against K.pneumoniae, P.aeruginosa
and A. baumannii in pneumonia based on QID I.V.
administration
• Very good efficacy against K.pneumoniae in pneumonia
based on aerosol administration
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Excellent efficacy in UTI
ED50 for AA139 in the bladder
E. coli
K. pneumonia
7
log10 cfu/g bladder
6
5
4
3
ED50 1.7 mg/kg
2
-1.0
-0.5
0.0
0.5
1.0
1.5
log10 [AA139] mg/kg
Euprotec 2013
Confidential
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Very good activity of Arenicin against K.
pneumonia in a neutropenic pneumonia model
following aerosol admin
Klebsiella Pneumonia NCTC13442
Variant
log reduction
MIC
AA139
-3.89
1
Colistin
-1.75
1
Euprotec 2013
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Toxicological summary
•
•
•
•
•
Selective and specific for bacteria
Wide therapeutic window – a factor of 25
MTD level of 25 mg/kg versus ED 50 of 1 mg/kg
Adverse effects related to histamin release
Changes in proximal tubuli the only, dose dependent and
reversible pathological finding
• Changes in NGAL correspond with pathological kidney
findings
• No cardiotoxic effect
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Product profiles of Meropenem, Colistin and
Arenicin
Indications
Meropenem
Colistin
Arenicin
Pneumonia
Complicated urinary tract infections
Coverage
+++
+++
+++
+++
+++
+++
XDR E.coli
++
+++
+++
XDR P.aeruginosa
++
+++
+++
XDR A.baumannii
+
+++
+++
KPC K.pneumonia
-
+++
+++
Colistin G- Bacteria
-
-
+++
Oral
no
no
no
IV
yes
yes
yes
IT
no
yes
yes
(yes)
yes
(no)
Neurological
no
yes
no
Hypersensitivity
yes
yes
yes
yes
yes
yes
Administration
Adverse events
Renal/Hepatic
Miscellaneous
Bactericidal
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