Confidential Corporate presentation,October 2013.
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Transcript of Confidential Corporate presentation,October 2013.
Confidential
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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Confidential
Arenicin selection process
Variant library generation(~250.000 variants)
~40 AMP’s identified
Several G+ but only one G- identified !
> 500 organisms screened for antimicrobial activity
NZ17074
Second variant library (~90.000 variants)
1500 hits but only 10 variants selected
First Hit
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Clin Cand – AA139
Confidential
Mode of action summary
• Arenicin acts at least partly due to non-lipid A-mediated 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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Confidential
• 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 interacts with the lipid transporter protein MLAC
Confidential
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
6University of Queensland 2013
Arenicin causes loss of cell surface structure and partial cytoplasm clearing in E. coli
Confidential
Arenicin causes ATP release without dramatic changes in cell morphology
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.
0 16 64 256 1024 40960
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10
15
20
25
Arcolpip
Extracellular ATP after 10 min
x MIC
Fold
change
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.
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Novozymes A/S, 2010
Confidential
Arenicin and Colistin have different effects on osmotic shock genes
Symbol Gene ID
Colistin µg/ml
Arenicin µg/ml
Description2½ 2½ 5 10
Osmotic shock genesosmB B1283 63 29 8 3 lipoprotein, osmotically inducible,osmC B1482 10 - - - PeroxiredoxinosmY B4376 5 - - - Osmotically-inducible protein Y precursorosmE B1739 3 - - - Osmotically-inducible lipoprotein E precursorbdm B1481 63 11 6 3 Biofilm-dependent modulation proteinosmotically inducible
sra B1480 11 4 3 - rpsV, osmotically induciblercsA B1951 6 2 - - positive regulator for ctr capsule biosynthesis,
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Novozymes A/S, 2010
Whole Genome TraDIS preliminary data suggest inhibition of key cytosolic processes in gene translation
Confidential
In vitro efficacy summary
• 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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Confidential
# 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
Potent in vitro activity against GAIN pathogens
MIC90 determinations (MDR clinical isolates)
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Eurofins medinet 2012
Confidential
Extremely low spontaneous mutational frequency to Arenicin
Organism Isolate ID
Resistance Frequency
(4XMIC)
Resistance Frequency
(8XMIC)
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
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Eurofins medinet 2012
Confidential
NBS plates- MHB only
Compound ID
GN_01E.coli
ATCC 25922
GN_03K. pneumoniaeATCC 700603
GN_04K. pneumoniae
ATCC 13883
GN_34A.baumanniiATCC 19606
GN_42P.aeruginosaATCC 27853
GN_43P.aeruginosaPolymixin R
GN_44 E.coli
ATCC 10536
GN_45K. pneumoniae
BAA_2146
MIC [μg/mL]
Colistin MCC_000094B ≤0.03 ≤0.03 ≤0.03 0.03/0.06 1/2 32/64 0.03/0.125 0.06
Ciprofloxacin MCC_000166 ≤0.03 0.25 ≤0.03 0.5/1 2 0.25/0.5 32/64 >64
AA139 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 MIC [μg/mL] Colistin
MCC_000094B 0.06 1/0.5 0.125 0.5/0.25 0.5 64 0.5/0.25 0.125
Ciprofloxacin MCC_000166 ≤0.03 2 0.5 >64 1 1 2/1 >64
AA139 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
Limited effect of mucin (Survanta) on in vitro
efficacy
University of Queensland 2013
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Confidential
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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Confidential 14Euprotec 2013
PK/PD dose fractionation study shows that Cmax drives the efficacy of
Arenicin (NZ17230)(5mg/kg over 3 days seems to exert maximal effect)
Q36
h
Q18
h
Q12
h
Q8h
Pre-Treat-ment
NZ17230 - 3mg/kg
NZ17230 - 3mg/kg
NZ17230 - 3mg/kg
NZ17230 - 3mg/kg
1.00E-01
1.00E+00
1.00E+01
1.00E+02
1.00E+03
1.00E+04
1.00E+05
1.00E+06
1.00E+07
Tissue burdens following infection with E. coli UT189
KidneyBladderUrine
CFU
/mL
hom
ogen
ate
Confidential
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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Confidential
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Euprotec 2013
ED50 for AA139 in the bladder
Excellent efficacy in UTI
E. coli K. pneumonia
log10 [AA139] mg/kg
log
10 c
fu/g
bla
dd
er
-1.0 -0.5 0.0 0.5 1.0 1.52
3
4
5
6
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ED50 1.7 mg/kg
Confidential
Very good activity of Arenicin against K. pneumonia in a
neutropenic pneumonia model following aerosol admin
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Klebsiella Pneumonia NCTC13442
Variant log reduction MIC
AA139 -3.89 1
Colistin -1.75 1
Euprotec 2013
Confidential
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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Confidential
Indications Meropenem Colistin ArenicinPneumonia +++ +++ +++Complicated urinary tract infections +++ +++ +++Coverage
XDR E.coli ++ +++ +++XDR P.aeruginosa ++ +++ +++XDR A.baumannii + +++ +++KPC K.pneumonia - +++ +++Colistin G- Bacteria - - +++Administration
Oral no no noIV yes yes yesIT no yes yesAdverse events
Renal/Hepatic (yes) yes (no)Neurological no yes noHypersensitivity yes yes yesMiscellaneous
Bactericidal yes yes yes
Product profiles of Meropenem, Colistin and Arenicin
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