Post on 26-Mar-2019
transcript
Adenium Biotech
Management:
- Peter Nordkild, MD, CEO, ex Novo Nordisk, Ferring, Egalet
- Søren Neve, PhD, project director, ex Lundbeck, Novozymes
Board of Directors:
- Stephan Christgau, PhD, chairman, Novo A/S
- Anker Lundemose, MD, ex Novo Nordisk, ex OSI Pharmaceuticals
- Khalid Islam, PhD, ex Arpida
- Ejner Bech Jensen, MSc, VP R&D Novozymes A/S
Current Investor:
- Novo A/S
Bad Bugs – need new drugs
• The total US market of hospital acquired infections
was in 2006 estimated at USD 7.9 billion.
• 65% of hospital acquired infections are caused by
Gram-negative bacteria (Clin Infect Dis 2005;41:848–854)
Resistance
Resistance is inevitable and requires novel antibiotics
Antibiotic Year deployed Resistance observed
Sulfonamides 1930s 1940s
Penicillin 1943 1946
Streptomycin 1943 1959
Chloramphenicol 1947 1959
Tetracycline 1948 1953
Erythromycin 1952 1988
Vancomycin 1956 1988
Methicillin 1960 1961
Ampicillin 1961 1973
Cephalosporins 1960s Late 1960s
Arenicin program highlights
• Spin out from Novozymes’ AMP group
• Novel mode of action, no cross resistance to existing antibiotics
• Bactericidal on broad range of multidrug resistant Gram-negative
bacteria
• Development plan includes orphan drug opportunity
• Few competing bactericidal products in development
• Strong lead/back up product candidates
• Stable IV formulations
• Strong IP (2025-2030)
• Addresses significant unmet Gram-negative clinical need
• Large, growing and non-generic hospital market of USD 8 billion
• Hospital and primarily ICU based specialist target group requiring
small sales force
ESKAPE Pathogens
• Enterococcus faecium
• Staphylococcus aureus
Klebsiella species
Acinetobacter baumannii
Pseudomonas aeruginosa
Enterobacter
E.coli
E. aerogenes
E. cloacae
Arenicin-3 NZ17000
• Isolated from lugworm (Arenicola marina)
• 21 amino acids
• Very stable beta-hairpin structure
• MW 2.613 kDa
• pI ~ 11.27
1. screen – S. cerevisiae (250.000)
- plate screen vs. E. coli in the
presence of 5% blood
- retest vs. other Gram- bacteria
2. screen – purified peptide (250)
- MICs vs. 11 bacteria +/- serum
- killing kinetic
- protein binding
3. screen – chemical synthesis (10)
- a range of in vitro & in vivo
parameters
- MTD, peritonitis, cystitis etc
2 new Arenicin variants
• NZ17143 and NZ17211 with distinct features
• Improved protein binding properties
• Improved toxicological properties
Output Evaluation of variants
Variant generation through consecutive
screening of yeast libraries
250.000 variants
Antimicrobial spectrum and protein binding
of NZ17000, NZ17143 and NZ17211
Arenicin variant
E. coli (μg/ml)
K. pneumoniae (μg/ml)
E. cloacae (μg/ml)
P. aeruginosa (μg/ml)
S. maltophilia (μg/ml)
A. baumanii (μg/ml)
M. catarrhalis (μg/ml)
Protein Binding
NZ17000 0.25 1 1 0.5 0.5 0.5 0.5 99
NZ17143 0.25 4 4 2 8 4 2 85
NZ17211 0.5 8 8 4 8 4 2 80
Antimicrobial activity against selected
resistant Gram-negative strains
Bacteria No NZ17000 NZ17143 NZ17211
MIC (µg/ml) MIC (µg/ml) MIC (µg/ml)
Carbapenem resistant
Klebsiella pneumoniae
(KPC2)
2 1 2 4
Carbapenem resistant
Pseudomonas
aeruginosa
1 1 4 4
Carbapenem resistant
Klebsiella pneumoniae
(NDM-1)
1 2 2 2
Tigecycline resistant
Salmonella Hadar
1 1 1 1
MoA - localization of Arenicin
45 min 30 min 15 min
Control
Fosfomycin
Polymycin B
Arenicin-3
A. E. coli exposed for 30 min to NZ17000 and stained with TRITC. Treatment with NZ17000 results in influx of TRITC into the E. coli
B. E. coli exposed for 30 min with TRITC labelled NZ17000. Clusters of NZ17000 were localized in the bacterial membrane
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.
MoA - ATP efflux after treatment
with Arenicin
0
5
10
15
20
25
0 16 64 256 1024 4096
Ar
col
pip
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.
Arenicin:
Perturbs the membrane
potential increasing the
permeability of the bacterial
membrane
Inhibits the protein synthesis
Time kill of NZ17000 vs E.coli and
Pseudomonas
Time kill kinetics for Escherichia coli ATCC25922
NZ17000 in ca-Müller Hinton broth
0 30 60 90 120 150 180101
102
103
104
105
106
107
108
109
Growth control
10 x MIC Gentamicin (5µg/ml)10 x MIC Arenicin-3 (5µg/ml)
Time (minutes)
CF
U/m
l
NZ17000 Time killing kinetics forPseudomonas aeruginosa ATCC27853
0 30 60 90 120 150 180101
102
103
104
105
106
107
108
Growth Control0.25 x MIC NZ17000 ( 0.125µg/ml)2 x MIC NZ17000 (1µg/ml)
10 x MIC NZ17000 (5µg/ml)
Time (minutes)
CF
U/
ml
of
Pseu
do
mo
nas
ED50 ~1.5 mg/kg in urine and ~ 1.8 mg/kg in the bladder
Dose response of NZ17000 in the UTI mouse model
Dose response of NZ17143 in the UTI
mouse model
ED50 < 0.8 mg/kg in urine, bladder and in the kidneys
Dose response of NZ17211 in the UTI
mouse model
ED50 < 0.8 mg/kg in urine, bladder and in the kidneys
Dose response in neutropenic mouse peritonitis model
against a multi-resistant E. coli at 5 hours after treatment
Blood Peritoneal fluid
NZ1
71
43
N
Z17
21
1
NZ1
70
00
Efficacy of NZ17143/NZ17211 against multi-resistant E. coli in
the murine Urinary Tract Infection model (UTI)
vehic
le
NZ1
7143
12.
5 m
g/kg
NZ1
7211
20
mg/
kg
Mer
openem
40
mg/
kg
2
3
4
5
6
7
8Urine day 2 post infection
Lo
g1
0C
FU
/ml
vehic
le
NZ17
143
12.5
mg/k
g
NZ17
211
20 m
g/kg
Mer
openem
40
mg/k
g
2
3
4
5
6
7
8Bladder day 3 post infection
Lo
g1
0C
FU
/ml
vehic
le
NZ17
143
12.5
mg/k
g
NZ17
211
20 m
g/kg
Mer
openem
40
mg/k
g
1
2
3
4Kidneys day 3 post infection
Lo
g1
0C
FU
/ml
Kidney 3 days post infection Bladder 3 days post infection Urine 2 day post infection
Pharmacokinetic properties
after IV administration
Arenicin-3
variant
Protein
binding
(%)
T½
(min)
AUC
(min*ug/ml)
Cmax
(ug/ml)
Bioavailability
(%, SC vs IV)
NZ17000 >99 130 532 7.7 12
NZ17143 85 69 324 7.5 70
NZ17211 80 60 432 9.9 60
Toxicological overview
of NZ17000, NZ17143 and NZ17211
Variant NZ17000
(mg/kg)
NZ17143
(mg/kg)
NZ17211
(mg/kg)
MTD (IV) 15 25 30
NOAEL (IV) 7,5 15 20
Arenicin summary
• New mode of action
• Spontaneous mutational frequency for E. coli is 3X10-9 and P.
aeruginosa >10-8
• Potent in vitro activity against a wide spectrum of Gram-
negative bacteria
• Rapidly bactericidal – MBCs ~ MICs
• No cross resistance to known antibiotics
• No or little inoculum effect
• Favorable efficacy in experimental animal models of infection
– Septicemia against E. coli and P. aeruginosa
– UTI against E. coli
– Thigh infections against E. coli.