The Zinchel Project

Oslo, May 2016

Pål Rongved

The ZinChel Project - Overcoming β-Lactam

Resistance

Background, results and plans

Contents:

• Background: the post-antibiotic era – why are we loosing?

• What is the ZinChel technology?

• Results

• Specific tasks to investigate resistance and mechanism of action

• Plans and budget

• The link to HORIZON2020 and other EU initiatives

• People and project team

• Summary

Background

Bacteria – the first cells on earth?

Ancient Fossil Bacteria : Pictured above are two kinds cyanobacteria from

the Bitter Springs chert of central Australia, a site dating to the Late

Proterozoic, about 850 million years old. On the left is a colonial

chroococcalean form, and on the right is the filamentous Palaeolyngbya.

Sources: Bitter Springs chert fossil image provided by J. William Schopf. Image of stromatolites provided by the University of

Wisconsin Botanical Images Collection

• The oldest known fossils: cyanobacteria from Archaean rocks of

western Australia, dated 3.5 billion years old.

• The oldest rocks: a little older: 3.8 billion years old!

• Milton Mainwright, Sheffield University “Imhokep, the notable ancient

Egyptian healer, (….) is known to have treated surface infections with

mouldy bread“,

• In 1874: Sir William Roberts: cultures of the mold with Penicillium

glaucum was antibacterial.

Gorgonzola,

an Italian

cheese

containing

"veins" of

Penicillium

glaucum

Statuette of Imhotep

in the Louvre

Drug Discovery within Antibiotics (AB)

Penicillinase: bacterial weapon destroying the AB

1940 – first penicillinase discovered

1942 – first penicillin became «available»

Carbapenems:

One of the last resort AB’s today The first one, Imipenem. FDA approved

1985.

Metallo-β-lactamases (MBL): new, more

efficient new penicillinases: first crystal

structure solved in 1995

Alexander Fleming: penicillin G in 1928

- the 1945 Nobel Price in medicine

Silver LL. Clin. Microbiol. Rev. 2011

CDC 2013

Ampicillin, Piperacillin-tazobactam, Mecillinam, Cefotaxim,

Ceftazidime, Meropenem, Imipenem, Ciprofloxacin, Levofloxacin,

Gentamicin, Tobramycin, Azitromycin, Tigecyline, Colistin,

Fosfomycin, Trimetoprim-sulfamethoxazole, Tetracycline,

Chloramphenicol, Temocillin, Nitrofurantoin, Amoxicillin, Oxacillin,

Ticarcillin, Cefepime, Doripenem, Aztreonam, Ceftobiprole,

Cefalexin, Ertapenem, Amikacin, Netilimicin, Vancomycin,

Telavancin, Norfloxacin, Erytromycin, Clindamycin, Minocycline,

Daptomycin, Spectinomycin…….etc…..etc….etc….etc…..

Why are we loosing?

• Polymyxin (colistin) resistance: singularly due to the plasmid-carried mcr-1 gene.

• Readily transmissable

• Plasmid transfer possible also to Klebsiella pneumoniae and Pseudomonas aeruginosa

• mcr-1 encodes a phosphoethanolamine transferase (zinc dependent enzymes)

• “Findings emphasize the urgent need for coordinated global action in the fight against pan-drug-resistant Gram-negative bacteria”

China autumn 2015: the fall of a last resort antibiotic The mcr-1 gene for polymyxin resistance on the run..

Liu et al, Lancet Infect Dis 2015

Horizontal gene transfer

between different species

• The most threatening bacteria today: the

Gram negatives

• In 30 years: only two genuinely first-in-class

AB to the market (US)

• None against Gram negatives!

What new Antibiotics (AB) are marketed?

Dalbavancin

(EU: Xydalba, Actavis)

Newly discovered:

Teixobactin Ling et al, Nature L517 (2015) 455

The ZinChel project Discovery and results

The ZinChel project – a new strategy against resistant bacteria

Based on chemistry – rational design

How penicillins work: Mechanism of action (MOA) β-lactams:

Our approach:

How carbapenemases work: ZINC!

PCT Examiners

patentability report 2015:

No-one else is working

with the same:

Results

ZinChel Results – Testing Clinically Relevant Resistant Bacteria (Ø. Samuelsen, UNN)

ZN41

IC50: 7,7µM

ZN53

IC50: 1,8µM

ZN58

IC50: 0,4µM Activity against

pure enzyme

(VIM-2)

Results Clinical Isolates

ID, bacterial strain

MIC (mg/L)

K34-7 K66-45

Name, bacterial strain/cell type

P. aeruginosa K.

pneumoniae

Inhibitor concentration: 125 µM

β-Lactamase VIM-2 NDM-1

MEM 32-64 32

ZN74 >1000 >1000

ZN110 >1000 >1000

TPEN >1000 500

MEM + ZN74 2 ≤ 0,125

MEM + ZN110 2 0,5

MEM + TPEN 1 ≤ 0,5

MEM + Captopril 32 64

Meropenem

(MEM) Captopril

Results Clinical Isolates

ID, bacterial strain

MIC (mg/L) IC50 (µM)

K34-7 K66-45 Human Cancer Cells (µM) Healthy human

cells

Name, bacterial strain/cell type

P. aeruginosa K.

pneumoniae

Breast cancer Pancreas cancer

hepG2 Inhibitor concentration: 125µM MDA-MB-231 MiaPaCa Colo357

β-Lactamase VIM-2 NDM-1 n.a. n.a. n.a. n.a.

MEM 32-64 32 n.a. n.a. n.a. n.a.

ZN74 >1000 >1000 4.9 ± 0.7 4.8 ± 1.2 6.8 ± 1.4 ~ 100

ZN110 >1000 >1000 101 ± 21 66 ± 13 112 ± 33 >> 100

TPEN >1000 500 2.5 ± 0.7 3.5 ± 2.2 2.9 ± 1.1 ~ 10

MEM + ZN74 2 ≤ 0,125 n.a. n.a. n.a. n.a.

MEM + ZN110 2 0,5 n.a. n.a. n.a. n.a.

MEM + TPEN 1 ≤ 0,5 n.a. n.a. n.a. n.a.

MEM + Captopril 32 64 n.a. n.a. n.a. n.a.

Meropenem

(MEM) Captopril

Results Clinical Isolates

ID, bacterial strain

MBL-positive Gram-negatives

P. Aeruginosa (K34-7) – VIM-2 K. Pneumoniae (K66-45) - NDM1

Conc. inhibitor(µM) 50 31,3 15,6 50 31,3 15,6

MEM+ZN141 1 1 32 0,125 0,125 16

MEM+ZN142 1 2 16 0,125 0,125 8

MEM+ZN144 1 2 32 0,125 0,125 16

MEM+ZN145 1 1 4 0,25 0,125 4

MEM+ZN147 2 2 32 0,125 0,125 16

MEM+ZN148 1 1 16 0,125 0,125 8

MEM+ZN155 1 4 32 0,125 4 16

MEM + TPEN 1 ≤ 0,5 n.a. n.a. n.a. n.a.

MEM + Captopril

32 64 n.a. n.a. n.a. n.a.

Meropenem

(MEM)

Captopril

Results - hematology

Tromsø, February 2016:

No colorization in red blood

cell suspensions in

concentretions of lead

candidates < 500 µM

Jim O’Neill, 2016:

Ongoing work

Microbe-targeted

Zn-chelators

(as for other AB

projects)

Toxicity?

• Cell lines

• Animal

models

Efficacy?

Resistance

development?

Mechanism of

action? • β-lactamases?

• PBPs?

• Other Zn-dependent

enzymes?

• Zn-homeostasis?

Project plan

Dr Ørjan Samuelsen

Dag Berild, et al OUS

Opportunities

Horizon2020 – IMI – ND4BB ZinChel Spring 2015: open invitation into the IMI programme

ENABLE

• IMI: the world's biggest

public-private partnership

(PPP) in the life sciences.

• IMI 2 programme: €3.3

billion budget 2014-2024.

Of this:

• €1.638 billion comes from

Horizon 2020

• €1.425 billion is committed

to the programme by

EFPIA companies

• Up to €213 million can be

committed by other life

science industries or

organisations that decide

to contribute to IMI 2 as

members or Associated

Partners in individual

projects

Horizon2020

Summary

• Only two genuinely new classes of antibiotics in 30 years

• Industry is reluctant because of rapid development of resistance.

• ZinChel: genuinely new adjuvant technology, dramatically reducing resistance.

• ZinChel make the carpapenems efficient again (resistant clinical isolates).

• The scope of the ZinChel approach: very wide.

• Not based on natural products but on medicinal chemistry rational design.

• Key studies ongoing: resistance potential and in vivo tox.

• An open door to EU through the IMI/Enable programme.

People

Associate Professors

Annette Bayer

Dr Hanna-Kirsti Leiros

Postdoc Zeeshan Mohamad

Dr Ørjan Samuelsen

Postdoc Silje Lauksund

O. Alexander H. Åstrand

Scientist Geir Kildahl-Andersen

Scientist Christian Schnaars

PhD student Elvar Ø. Viktorsson

MSc student Ørjan Apeland

PhD student Anthony Prandina

Associate Professor Lars Petter Jordheim

Funding:

Center for Integrative

Microbial Evolution, UiO

CIME PhD

student

Anthony

Prandina

Centre for Integrative

Microbial Evolution

(CIME), UiO

CIME PhD student

Anthony Prandina

Hanne Winther-Larsen

Mike Koomey

Tom Kristensen

Dirk Linke

Ole Andreas Økstad

People

26

Thank you for your attention…