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

Key antibacterial needs 2011

Source: Datamonitor 02/2011.

Arenicin

A new Gram-negative antibiotic

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

Healthcare associated infections in the US 2010

Source: Datamonitor 02/2011

Nosocomial indications - volume/value

Source : Datamonitor 03/2007

ESKAPE Pathogens

• Enterococcus faecium

• Staphylococcus aureus

Klebsiella species

Acinetobacter baumannii

Pseudomonas aeruginosa

Enterobacter

E.coli

E. aerogenes

E. cloacae

Generation of new variants

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.