It all gone to worms !
The role of the odilorhabdins!
NOSOPHARM – BSAC Spring Conference 2019 – March 2019 THE BLACK SWANS OF RESISTANCE
x6 over 6 years! +35%/year
NOSOPHARM – BSAC Spring Conference 2019 – March 2019 2 ARE WE PREPARED FOR THE NEXT BLACK SWANS?
Carbapenem resistance in K. pneumoniae, Italy 2006-2017
40 35 30 25 20 15 resistance 10 % 5 0 2006 2007 2008 2009 2010 2011 2012 2013 2014 2015 2016 2017 [Source: ECDC/EARS-Net]
Preparedness lies in the diversity of the anti-infective arsenal
NOSOPHARM – BSAC Spring Conference 2019 – March 2019 3 A DIVERSE BUT AGEING ANTIBIOTIC ARSENAL
Gram(+) Glycopeptides Gram(-) Macrolides Sulfonamides Aminosides Streptogramins Fidaxomicins
1930 1940 1950 1960 1970 1980 1990 2000 2010 2020
β-Lactams I β-Lactams II β-Lactams III Oxazolidinones (penicillins) (cephalosporins) (carbapenems) Lipopeptides Tetracyclines Quinolones Polymyxins Trimethoprims Phenicols Rifamycins
NOSOPHARM – BSAC Spring Conference 2019 – March 2019 4 MICROBIAL DIVERSITY: A PROLIFIC SOURCE OF ANTIBIOTICS
12 Microbial origin Glycopeptides 5 Synthetic origin Macrolides Sulfonamides Aminosides Streptogramins Fidaxomicins
1930 1940 1950 1960 1970 1980 1990 2000 2010 2020
β-Lactams I β-Lactams II β-Lactams III Oxazolidinones (penicillins) (cephalosporins) (carbapenems) Lipopeptides Tetracyclines Quinolones Polymyxins Trimethoprims Phenicols Rifamycins
NOSOPHARM – BSAC Spring Conference 2019 – March 2019 5 THE ACTINOMYCETES: AN OVEREXPLOITED FIELD
>10 millions strains of Actinomycetes need to be screened to discover a new antibiotic
NOSOPHARM – BSAC Spring Conference 2019 – March 2019 6 PHOTORHABDUS & XENORHABDUS: A NEW LODE
Photorhabdus & Xenorhabdus are bacterial symbionts of the entomopathogenic nematodes Heterorhabditis & Steinernema γ-proteobacteria (Enterobacterales) Genome size: ~4.5 Mb Started to be investigated in the 1980’s ~300 species/subspecies described
NOSOPHARM – BSAC Spring Conference 2019 – March 2019 7 ECOLOGICAL RATIONALE
Photorhabdus or Xenorhabdus (green) live within the intestine of the host nematode
The host nematode infects an insect
Photorhabdus or Xenorhabdus are released within the insect and produce compounds to kill the insect
Photorhabdus or Xenorhabdus produce a set of antibiotic molecules to prevent microbial competitors of the environment to degrade the corpse of the insect
The nematode and the bacteria use the biomass of the insect as nutrients to reproduce. Photorhabdus or Xenorhabdus colonise the nematode who will emerge from the corpse of the insect
NOSOPHARM – BSAC Spring Conference 2019 – March 2019 8 GENOMIC RATIONALE
Streptomyces bingchenggensis Streptomyces bingchenggensis Streptomyces griseus Xenorhabdus sp. #6 Nocardia brasiliensis Xenorhabdus nematophila Streptomyces cattleya Xenorhabdus sp. #5 Streptomyces avermitilis Xenorhabdus sp. #3 Stigmatella aurantiaca Xenorhabdus sp. #4 Nostoc punctiforme Streptomyces griseus Xenorhabdus sp. #6 Xenorhabdus sp. #2 Micromonospora aurantiaca Xenorhabdus bovienii Xenorhabdus sp. #5 Nocardia brasiliensis Xenorhabdus nematophila Salinispora arenicola Xenorhabdus sp. #4 Photorhabdus luminescens Nocardia cyriacigeorgica Nocardia cyriacigeorgica Salinispora arenicola Streptomyces avermitilis Xenorhabdus sp. #3 Photorhabdus asymbiotica Photorhabdus luminescens Micromonospora aurantiaca Nocardia farcinica Nocardia farcinica Xenorhabdus sp. #2 Nostoc punctiforme Burkholderia thailandensis Burkholderia thailandensis Xenorhabdus bovienii Mycobacterium bovis Photorhabdus asymbiotica Salinispora tropica Pseudomonas fluorescens Pseudomonas fluorescens Streptomyces scabiei Burkholderia mallei Pseudomonas syringae pv. Tomato Pseudomonas syringae pv. Tomato Salinispora tropica Streptomyces cattleya Burkholderia mallei Xenorhabdus sp. #1 Streptomyces coelicolor Microcystis aeruginosa Mycobacterium bovis Streptomyces scabiei Anabaena variabilis Anabaena variabilis Microcystis aeruginosa Pectobacterium atrosepticum Ralstonia solanacearum Stigmatella aurantiaca Nostoc sp Streptomyces coelicolor Pectobacterium atrosepticum Nostoc sp Pseudomonas aeruginosa PKS & NRPS Escherichia coli Domains/Mb Xenorhabdus sp. #1 Domains Pseudomonas aeruginosa Sinorhizobium medicae Ralstonia solanacearum Escherichia coli Sinorhizobium medicae 0 100 200 300 400 500 600 700 0 10 20 30 40 50 60 NOSOPHARM – BSAC Spring Conference 2019 – March 2019 9 DISCOVERY OF THE ODILORHABDINS
150 90 380 15 Strains antimicrobial antibacterial new antibacterial extracts selected fractions molecules
Odilorhabdins
69% with 1-9 antibacterial antibacterial activity fractions/extract (58% against G[-]) (median: 4)
NOSOPHARM – BSAC Spring Conference 2019 – March 2019 10 NOSO-95: THE FIRST DISCOVERED ODILORHABDINS
Molecule R1 R2 MW (g/mol) NOSO-95A -OH -OH 1 296.5 NOSO-95B -OH -H 1 280.5 NOSO-95C -H -H 1 264.5
NOSOPHARM – BSAC Spring Conference 2019 – March 2019 11 NOSO-502: FIRST ODILORHABDIN CLINICAL CANDIDATE
NOSO-502
2010: Screening of a collection of strains of Photorhabdus and Xenorhabdus 2011: Discovery of the Odilorhabdins 2012: Total synthesis achieved 2013-2017: Lead optimization by medicinal chemistry (650 chemical analogs) 2017: Selection of NOSO-502 as 1st odilorhabdin clinical candidate
NOSOPHARM – BSAC Spring Conference 2019 – March 2019 12 ODILORHABDINS INHIBIT THE RIBOSOME WITH A NEW MODE OF ACTION
o Co-crystallography studies showed Odilorhabdins bind on the bacterial ribosome at a site not exploited by any known ribosome-targeting antibiotic C D o Odilorhabdins induce miscoding, likely by increasing the affinity of aa-tRNAs to the ribosome
NOSOPHARM – BSAC Spring Conference 2019 – March 2019 13 ANTIBACTERIAL SPECTRUM OF NOSO-502
NOS: NOSO-502 CIP: Ciprofloxacin GEN: Gentamicin IPM: Imipenem TGC: Tigecycline PMB:Polymyxin B
NOSOPHARM – BSAC Spring Conference 2019 – March 2019 14 ANTIBACTERIAL ACTIVITY AGAINST CARBAPENEM-RESISTANT ENTEROBACTERIACEAE
NOS: NOSO-502 CIP: Ciprofloxacin GEN: Gentamicin IPM: Imipenem TGC: Tigecycline PMB:Polymyxin B
EUCAST MIC breakpoints 2018 Red: Resistant Orange: Intermediate Green: Susceptible
NOSOPHARM – BSAC Spring Conference 2019 – March 2019 15 KINETICS OF BACTERICIDALITY
E . c o li A T C C 2 5 9 2 2 K . p n e u m o n ia e A T C C 4 3 8 1 6 1 0 1 0 9 9
8 8
l
l m
m 7 7
/
/ U
U 6 6
F
F C
C 5 5
0
0 1
4 1 4
g
g o
3 o 3
L L 2 2 1 1 0 0 0 2 4 6 8 1 0 1 2 1 4 1 6 1 8 2 0 2 2 2 4 0 2 4 6 8 1 0 1 2 1 4 1 6 1 8 2 0 2 2 2 4 T im e (h ) T im e (h )
P o s itiv e c o n tro l N O S O -5 0 2 4 xM IC N O S O -5 0 2 8 xM IC
NOSOPHARM – BSAC Spring Conference 2019 – March 2019 16 SPONTANEOUS FREQUENCY OF RESISTANCE
Strain MIC (µg/mL) FoR (4xMIC) FoR (8xMIC)
E. coli ATCC 25922 4 3.0E-9 <5.2E-10
K. pneumoniae ATCC 43816 1 2.4E-9 <7.0E-10
NOSOPHARM – BSAC Spring Conference 2019 – March 2019 17 EFFICACY IN PERITONITIS/SEPSIS MODELS
Experimental model: peritoneal infection of neutropenic mice (3 mice/group) at t0 with E. coli EN122 (ESBL); antibacterial administration at t0+1h (SC); bacterial enumeration in blood at t0+5h; colistin used as positive control (not comparator); One-way ANOVA, Dunnett’s comparison vs control T26h group, ns: not significant, *: p≤0.05, **: p≤0.01, ***: p≤0.001, ****: p≤0.01
8
n s d
o 6 NOSO-502 shows significant burden
o n s
l
b
l reductions in blood: m / **** v
U 4 1 log10 CFU/ml @ 2.6 mg/kg F
C **** **** **** **** v 2 log10 CFU/ml @ 3.8mg/kg
0 1
g 2 v 3 log10 CFU/ml @ 9.8mg/kg
o L
0
t e ) ) ) ) ) ) ) n l g g g g g g g c e i /k /k /k /k /k /k /k m h g g g g g g g t e a m m m m m m m e V r 3 5 5 0 0 0 5 t . . ( 1 2 4 ( f 1 2 ( ( ( ( ( 2 n o 0 2 2 2 ti t 2 2 5 0 0 0 r 0 0 - is 5 5 -5 -5 -5 l ta - - O o S O O S O O O C S S O S S S O O N O O O N N N N N
NOSOPHARM – BSAC Spring Conference 2019 – March 2019 18 EFFICACY IN URINARY TRACT INFECTION MODEL
Experimental model: trans-urethral infection of mice (C3H/HeN; 8 mice/group) at t0 with E. coli UTI89; administration of NOSO-502 (NOS) at t0+24h, t0+48h and t0+72h (SC); bacterial enumeration in bladder and kidney tissue at t0+96h; ciprofloxacin (10 mg/kg BID, IV) used as positive control (not comparator). Statistical result for 96 h burden compare to vehicle. Kruskal-Wallis statistical test of terminal burden at day 4 post infection (corrected for multiple comparisons, StatsDirect-Conover-Inman). ns: not significant, *: p≤0.05, **: p≤0.01, ***: p≤0.001, ****: p≤0.0001
K id n e y B la d d e r U rin e 8 1 0 1 0
e n s n s
e * * u n s u
e n s s
s * * * * *
n s
s 8 8
i
i
i
t t
6 * * * * r
* *
f
f
u
o
o
l
6 6
m
m
m
/
a
a
r r
4 U
g * * * *
g * * * *
F
/
/
U U
4 C 4
F
F
0
1
C
C
0 0
2 g
1 1
2 o 2
g
g
L
o
o
L L 0 0 0 t ) ) ) ) e t ) ) ) ) t ) ) ) ) n l g g g D n le g g g D n le g g g D e c k k k I e k k k I e k k k I i / / / B ic / / / ic / / / m g g g B B t h , tm h g g g , tm h g g g , a e m m m g a e m m m g a e m m m g e v k v v r 4 2 4 / e 4 2 4 /k e 4 2 4 /k t ( 1 2 g tr ( tr ( f ( ( (1 (2 g (1 (2 g 2 m f 2 f 2 o 0 2 2 o 0 2 2 m o 0 2 2 m t 5 0 0 0 t 5 0 0 0 t 5 0 0 0 r - 5 5 1 r - r - a - - ( -5 -5 (1 -5 -5 (1 t O ta O ta O S S O O n S O O n S O O n S S i S i S i O c O S S c O S S c N O O a N O O a N O O a N N x N N x N N x lo o o f fl fl o o o r r r p i ip ip C C C
NOSO-502 shows significant burden reductions in kidney (1.36 log10 CFU/g), bladder (1.96 log10 CFU/g), and urine (2.39 log10 CFU/ml) @ 24 mg/kg
NOSOPHARM – BSAC Spring Conference 2019 – March 2019 19 EFFICACY IN RESPIRATORY INFECTION MODEL
Experimental model: lung infection model of neutropenic mice (8 mice/ group) at t0 with MDR K. pneumoniae NCTC 13442 (OXA-48); antibacterial administration by subcutaneous route at t0+2h (q24h group) or at t0+2h, t0+8h, t0+14h and t0+20h (q6h groups); bacterial enumeration of lung tissue at t0+26h; One-way ANOVA, Dunnett’s comparison vs control T26h group, ns: not significant, *: p≤0.05, **: p≤0.01, ***: p≤0.001, ****: p≤0.01
e 1 0
u
s
s
i
t
g ****
8
n u
l **** **** **** ****
f
o NOSO-502 shows significant burden
m 6
a reductions in lung:
r g / v 2.69 log CFU/g @ 2 mg/kg q6h
U 10
F 4
C v 3.99 log10 CFU/g @ 6 mg/kg q6h
0 1
g v 4.07 log10 CFU/g @ 20 mg/kg q6h o
L 2
t e ) ) ) ) ) n l h h h h h e ic 6 6 6 4 6 m h q q q 2 q t e , , , q , a V g g g , g e k k k g k r / / / k / t g g g / g f g o m m m m t m r (2 (6 0 0 a (2 0 (4 t 2 2 (8 S 0 0 2 e 5 5 0 2 n - - 5 0 li O O - 5 c S S O - y S O c O O S e N N O g N O i N T NOSOPHARM – BSAC Spring Conference 2019 – March 2019 20 NOSO-502: FIRST ODILORHABDIN CLINICAL CANDIDATE
l Inhibits the bacterial ribosome with a novel mode of action l Active against multidrug-resistant bacteria, including Carbapenem-Resistant Enterobacteriaceae (CRE) l Bactericidal agent with fast kinetics of bactericidality l Safe and efficacious in vivo (peritonitis/sepsis, UTI, RTI) l IND-enabling studies to start in 2020, First-In-Human clinical trials to start in 2021
NOSOPHARM – BSAC Spring Conference 2019 – March 2019 21 RELEVANT PUBLICATIONS
NOSOPHARM – BSAC Spring Conference 2019 – March 2019 22 ACKNOWLEDGMENTS
This work has received financial support from OSEO and Region Languedoc-Roussillon under grant agreement A1010014J and from DGA under grant agreement 122906117. The research leading to these results has also received support from the Innovative Medicines Initiative Joint Undertaking under grant agreement 115583, the resources of which are composed of financial contributions from the European Union Seventh Framework Program (FP7/2007-2013) and EFPIA companies in kind contribution. NOSOPHARM, Lyon & Nîmes, France Statens Serum Institut, Copenhagen, Denmark University of Uppsala, Sweden v Maxime Gualtieri v Carina Vingsbo Lundberg v Diarmaid Hughes v Emilie Racine University of Illinois at Chicago, USA v Douglas Huseby v Lucile Pantel v Alexander S. Mankin v Sha Cao v Jessica Houard v Yury S. Polikanov EVOTEC, Manchester, UK v Matthieu Sarciaux v Tanja Florin v Kirsty Holden v Marine Serri v Malgorzata Dobosz-Bartoszek v Peter Warn v André Aumelas Institut Charles Gerhardt, Montpellier, France University of Fribourg, Switzerland v Jean-Marc Campagne v Patrice Nordmann v Renata Marcia de Figueiredo v Anthony Demords v Camille Midrier DGIMI, INRA, Montpellier, France UMR MD1, Marseille, France v Alain Givaudan v Jean-Michel Bolla v Sophie Gaudriault v Christelle Cotteaux-Lautard v Anne Lanois NOSOPHARM – BSAC Spring Conference 2019 – March 2019 23