Immuron Limited · ASX:IMC
Immuron — Technical Presentation (September 2015)
45 slides from Immuron's technical presentation, Sep 25, 2015
Slide 1: IMM-529 for the prevention and treatment of Clostridium difficile infections
IMM-529 for the prevention and treatment of Clostridium difficile infections
Slide 2: Clostridium difficile
Clostridium difficile http://www.nlm.nih.gov/medlineplus/images/clostridiumdifficile.jpg • Gram positive, spore forming anaerobe • Major nosocomial pathogen • Pathogenic C. difficile strains produce toxins that cause diarrhoea and mediate gut damage • Resistant to most clinical antibiotics
Slide 3: Clostridium difficile
Clostridium difficile • Leading cause of infectious antibiotic-associated diarrhoea in hospitals worldwide • C. difficile only colonises the gut if the normal microbiota is disrupted • People at greatest risk include those on antibiotics, the elderly and immunocompromised patients • High rate of rel
Slide 4: C. difficile infection (CDI)
C. difficile infection (CDI) • Causes a spectrum of diseases collectively known as CDI: - Mild self-limiting diarrhoea - Pseudomembranous colitis (PMC) • May progress to toxic megacolon, sepsis, death normal gut PMC toxic megacolon S. Kirov, U Tas
Slide 5: C. difficile infection (CDI)
C. difficile infection (CDI) Centers for Disease Control and Prevention (CDC): ANTIBIOTIC RESISTANCE THREATS in the United States, 2013 C. difficile is listed as the number one antibiotic resistance threat to the US healthcare system.
Slide 6: C. difficile spores
C. difficile spores • Spore-forming ability and spore persistence are major problems • Infectious particle • Survival mechanism • Heat, ethanol and UV resistant – allows for persistence in hospitals
Slide 7: The infectious cycle of C. difficile
The infectious cycle of C. difficile Disruption to the gut microbiota by antibiotics 1 Ingestion of spores 4 Toxin production by vegetative cells 2 Germination of spores • Toxin A in the colon • Toxin B 5 Toxin-mediated damage 3 Colonisation by vegetative cells in the colon
Slide 8: C. difficile toxins
C. difficile toxins • Toxin A (308 kDa) & Toxin B (269.6 kDa) • Monoglucosyltransferases encoded on PaLoc (19.6 kb) tcdR tcdB tcdE tcdA tcdC PaLoc 1 kb TcdR = alternative sigma factor TcdE = putative holin-like protein TcdC = anti-sigma factor (negative regulator of toxin production) • Toxin B shown
Slide 9: Pathogenesis of C. difficile toxins
Pathogenesis of C. difficile toxins Inactivation of Rho GTPases Depolymerisation of actin filaments Cytoskeleton disruption Cell rounding and death Jank et al., 2007
Slide 10: The hypervirulent strains
The hypervirulent strains • Emergence of hypervirulent strains associated with an increase in disease incidence, severity and mortality • Epidemic strains (ribotype 027) have been isolated worldwide • Higher relapse rates • Increased virulence may be due to: – A deletion in tcdC – loss of negative r
Slide 11: C. difficile in hospitals
C. difficile in hospitals • 3% of healthy people and 20-40% hospitalised patients colonised with C. difficile • associated with short-stay hospitals • poor hospital practices • antibiotic stewardship • ability to form spores – major problem C. difficile spore image: JoanneWee – resistant to many cle
Slide 12: Current treatment of CDI
Current treatment of CDI 1) Mild disease - Discontinue use of antibiotics 2) Moderate/severe disease - Treat with metronidazole, vancomycin or fidaxomicin 3) A range of non-antibiotic treatments have also been tested: • Intravenous IgG antibodies • Monoclonal antibodies • Probiotics • Non-toxigenic
Slide 13: Faecal transplant therapy
Faecal transplant therapy • Prepared by blending and filtering a fresh donor stool • Is administered via the upper or lower gastrointestinal tract by nasogastric/duodenal tube, colonoscopy or enema. • Restore the diversity of the gut microbiota and reverse the dysbiosis of CDI • Concerns over the lo
Slide 14: New methods for the prevention and/or treatment of CDI are urgently required
New methods for the prevention and/or treatment of CDI are urgently required
Slide 15: IMM-529 Production
IMM-529 Production • IMM-529 is a natural product which is intended to prevent and treat C. difficile infections • IMM-529 contains high concentrations of specific antibodies that are predominantly IgG (86%), IgA (7%) and IgM (7%) • IMM-529 does not destroy the gut microbiota like antibiotic treatme
Slide 16: Advantages of IMM-529
Advantages of IMM-529 • Inexpensive • Antibodies survive transit through the stomach and remain functional in the large intestine (site of C. difficile infection and toxin production) • Technology platform already used for the prevention and treatment of other gastrointestinal diseases (Cryptosporid
Slide 17: Evaluation of IMM-529 for passive immunotherapy in the prevention and treatment of C. difficile infections
Evaluation of IMM-529 for passive immunotherapy in the prevention and treatment of C. difficile infections
Slide 18: Different markets for IMM-529
Different markets for IMM-529 • Prevention of initial disease • Treatment • Prevention of disease relapse
Slide 19: Vaccine targets
Vaccine targets Disruption to the gut microbiota by antibiotics Ingestion of spores Toxin production • Toxin A Germination of spores • Toxin B Infection Colonisation by vegetative cells
Slide 20: IMM-529 Products
IMM-529 Products • IMM-529B contains high levels of specific antibodies which have been generated against recombinant Toxin B • IMM-529S contains high levels of specific antibodies which target the very infectious C. difficle spores • IMM-529V contains high levels of specific antibodies which target
Slide 21: In vitro characterisation of C. difficile IMM-529 antibodies
In vitro characterisation of C. difficile IMM-529 antibodies (spore, vegetative cell and toxin B)
Slide 22: IMM-529S1 antibodies are cross-reactive with the exosporium layer from C. difficile spores
IMM-529S1 antibodies are cross-reactive with the exosporium layer from C. difficile spores exosporium 1 2 3 4 5 6 7 8 250 kDa Non-immune 1-KI (A+B+) 250 kDa 2-M7404 (A+B+) 3-VPI10463 (A+B+) IMM-529S1 #1 4-GE (A+B+) 5-MDU2992 (A-B+) 250 kDa 6-JGS6133 (A+B+) 7-AI35 (A-B+) IMM-529S1 #2 8-1470 (A-B+) IM
Slide 23: IMM-529V1 antibodies are cross-reactive with cell lysates from C. difficile vegetative cells
IMM-529V1 antibodies are cross-reactive with cell lysates from C. difficile vegetative cells Animal Human 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 75 kDa 50 kDa 37 kDa 25 kDa IMM-529V1 contains antibodies that are cross-reactive with vegetative cell whole cell lysates from a variety of human and anima
Slide 24: IMM-529B1 antibodies are cross-reactive with Toxin B from different C. difficile strains
IMM-529B1 antibodies are cross-reactive with Toxin B from different C. difficile strains 1 2 3 4 5 6 7 8 9 10 11 1-KI (A+B+) 2-M7404 (A+B+) 250 kDa 3-VPI10463 (A+B+) Non-immune 4-GE (A+B+) 5-MDU2992 (A-B+) 6-JGS6133 (A+B+) 7-AI35 (A-B+) 250 kDa 8-1470 (A-B+) IMM-529B1 #1 9-CD37 (A-B-) 10-Purified To
Slide 25: IMM-529B1 antibodies neutralise Toxin B from historical and hypervirulent strains
IMM-529B1 antibodies neutralise Toxin B from historical and hypervirulent strains C o m m e r c ia l p u r ifie d T o x in B H is t o r ic a l T o x in B H y p e r v ir u le n t T o x in B ( s t r a in V P I1 0 4 6 3 ) ( s t r a in 6 3 0 ) (s t r a in K I) **** *** **** **** *** **** 100 100 100 75
Slide 26: IMM-529B1 antibodies neutralise Toxin B from a historical strain
IMM-529B1 antibodies neutralise Toxin B from a historical strain Toxin neutralisation assays 50 µm 50 µm 50 µm Vero cells only Vero + Toxin B-Hist Vero + Toxin B-Hist + anti-Toxin B antibody 50 µm 50 µm Vero + Toxin B-Hist + Non-immune Vero + Toxin B-Hist + IMM-529B1 • Antibodies in IMM-529B1 neutra
Slide 27: IMM-529B antibodies neutralise Toxin B from a hypervirulent strain
IMM-529B antibodies neutralise Toxin B from a hypervirulent strain Toxin neutralisation assays 50 µm 50 µm 50 µm Vero cells only Vero + Toxin B-HV Vero + Toxin B-HV + Toxin B antibody 50 µm 50 µm Vero + Toxin B-HV + Non-immune Vero + Toxin B-HV + IMM-529B1 • Antibodies in IMM-529B1 cross-neutralise
Slide 28: In vivo characterisation of IMM-529
In vivo characterisation of IMM-529
Slide 29: The C. difficile mouse model
The C. difficile mouse model C57BL/6 mice 6–7 weeks IMM-529 administration Day -10 -3 -2 -1 0 Antibiotics in drinking water to C. difficile challenge induce susceptibility to C. difficile (103 spores) Monitor: • Weight loss • Physiological appearance • Activity • Diarrhoea
Slide 30: IMM-529S1 protects 40% of mice from C. difficile disease
IMM-529S1 protects 40% of mice from C. difficile disease % survival % weight loss *** x Non-immune control IgG N = 10 mice/group IMM-529S1 *** p = 0.0005 SEM x = mice culled
Slide 31: IMM-529B1 protects 80% of mice from C. difficile disease
IMM-529B1 protects 80% of mice from C. difficile disease % W e igh t (re la tive to d a y 0 ) % survival % weight loss 11 0 *** 10 0 90 80 70 0 12 2 4 3 6 4 8 6 0 7 2 84 9 6 T im e ( h o u rs ) Uninfected (N=11) *** p < 0.0001 No IgG (N=10) SEM Non-immune IgG (N=20) IMM-529B1(N=20)
Slide 32: IMM-529B1 protects mice from C. difficile disease (prophylaxis)
IMM-529B1 protects mice from C. difficile disease (prophylaxis) Uninfected Non-immune IgG IMM-529B1 • Mouse colonic tissue from uninfected mice or mice that were pre-treated with Non- immune or IMM-259B1 prior to C. difficile infection. Tissue was stained with Periodic Acid Schiffs (PAS)/Alcian Blue
Slide 33: A combination IMM-529 product protects 80% of mice from C. difficile disease (prevention)
A combination IMM-529 product protects 80% of mice from C. difficile disease (prevention) % survival % weight loss ** x Non-immune IgG N = 5 mice/group IMM-529B1/IMM529S1/IMM-529V1 (1:1:1) ** p = 0.0027 SEM x = mice culled
Slide 34: Treatment with IMM-529B1 post-infection protects 80% of mice from disease
Treatment with IMM-529B1 post-infection protects 80% of mice from disease 120 100 U n in f e c t e d ( n = 1 5 ) P e r c e n t s u r v iv a l 80 **** N o Ig G ( n = 1 0 ) N o n -im m u n e Ig G ( n = 1 5 ) 60 IM M - 5 2 9 B 1 ( N = 1 4 ) 40 V a n c o m y c in ( n = 1 0 ) 20 0 0 12 24 36 48 60 72 84
Slide 35: Summary
Summary 1) IMM-529S1 (prophylaxis) • Protected 40% of mice 2) IMM-529B 1(prophylaxis) • Protected 80% of mice 3) IMM-529 product combination (prophylaxis) • Mixture of IMM-529B1, IMM-529S1 and IMM-529V1 protected 80% of mice 4) IMM-529B1 (treatment) • Protected 80% of mice when administered 6 hours
Slide 36: IMP Manufacture & Process Development
IMP Manufacture & Process Development Pilot Scale Production • Immuron has partnered with Diary Innovations Australia Limited and CSIRO to develop a scalable method of manufacture for IMM- 529 for a Clinical Development Program • Three pilot scale production batched of IMM-529B2, IMM-529S2 and IMM-5
Slide 37: Specific-ELISAs for Pilot Scale Products
Specific-ELISAs for Pilot Scale Products IMM-529B2 IMM-529V2 2 .5 2.5 IMM-529V IMM-529B A b s o r b a n c e (4 5 0 n m ) N o n -i m m u n e A b s o r b a n c e (4 5 0 n m ) 2 .0 2 .0 N o n - i m m u n e IgG 1 .5 1 .5 1 .0 1.0 0 .5 0 .5 0 .0 0 .0 A n ti b o d y ti te r A n ti b o d y ti te r IMM-529S
Slide 38: IMM-529B2 protects 60% of mice from C. difficile disease
IMM-529B2protects 60% of mice from C. difficile disease 100 U n in f e c t e d ( n = 5 ) N o Ig G ( n = 5 ) P e r c e n t s u r v iv a l 80 N o n - im m u n e Ig G ( n = 5 ) 60 IM M 5 2 9 B 2 (n = 1 0 ) Data summary (% survival): IM M 5 2 9 B 2 1 : 3 ( n = 1 0 ) 40 IM M 5 2 9 B 2 1 : 1 0 ( n = 1 0 )
Slide 39: IMM-529 protects 80% of mice from C. difficile disease
IMM-529 protects 80% of mice from C. difficile disease 100 U n in f e c t e d ( n = 1 0 ) N o Ig G ( n = 1 0 ) P e r c e n t s u r v iv a l 80 N o n -im m u n e Ig G ( n = 1 5 ) 60 IM M 5 2 9 V 2 (n = 5 ) Data summary (% survival): IM M 5 2 9 S 2 (n = 1 0 ) 40 IM M 5 2 9 B 2 (n = 2 0 ) • Uninfected-
Slide 40: Proposed model to test efficacy of IMM-529 against disease relapse
Proposed model to test efficacy of IMM-529 against disease relapse • Disease relapse model already established at Monash University • Disease relapses occurs in all mice treated with vancomycin • No disease relapse is observed in mice treated with fidaxomicin • Will use this model to test the effica
Slide 41: IMM529: Study Design
IMM529: STUDY DESIGN A Phase I/II study of IMM529 in patients with chronically-relapsing Clostridium difficile infection • Part A: A Phase I/II, dose evaluation safety study in combination with SOC in C. difficile patients • Part B: A Phase I/II, dose evaluation study investigating the activity of I
Slide 42: Major Inclusion/Exclusion Criteria
MAJOR INCLUSION/EXCLUSION CRITERIA Inclusion criteria: • In-patients • 18-75 years; • More than [TBC] episodes of diarrhoea bacteriologically confirmed C. difficile infection within the preceding month • Ability to provide written informed consent Exclusion criteria: • Receiving (or have received wi
Slide 43: IMM529-1 Study: Part A
IMM529-1 STUDY: PART A A Phase I/II study of IMM529 in patients with chronically-relapsing Clostridium difficile infection Part A: A Phase I/II safety assessment in combination with SOC in C. difficile patients Objectives: (i) To determine the safety of IMM529 tid for 14 days in combination with SOC
Slide 44: IMM529-1 Study: Part B
IMM529-1 STUDY: PART B A Phase I/II study of IMM529 in patients with chronically-relapsing Clostridium difficile infection Part B: A Phase I/II study investigating the activity of IMM529 in combination with SOC in patients with chronically-relapsing C. difficile infection Objectives: (i) To determin
Slide 45: Acknowledgements
Acknowledgements Jerry Kanellos A/Prof Dena Lyras Nicky Konstantopoulos Dr Melanie Hutton Ashley Turner Bliss Cunningham Reza Moussakhani Grant Rawlin