Altermune Kary B. Mullis The α-gal epitope • • α-gal is NOT produced in humans, chimps, orangutan or gt -/mouse A strong anti-α-gal response is already ubiquitous in all humans.
Download ReportTranscript Altermune Kary B. Mullis The α-gal epitope • • α-gal is NOT produced in humans, chimps, orangutan or gt -/mouse A strong anti-α-gal response is already ubiquitous in all humans.
Altermune
Kary B. Mullis
The α-gal epitope
• • α-gal is NOT produced in humans, chimps, orangutan or gt -/ mouse A strong anti α-gal response is already ubiquitous in all humans
The anti-Gal response
0.1uM
IgG 2
Altermune Linker
Aptamer will bind to M2e alpha-gal epitope
The Altermune Linker
Spiegelmer Strategy Eulberg and Klussmann ChemBioChem 2003
Altermune linker attaches to pathogen
Altermune linker attracts alpha-Gal antibodies
M2 target of amantadine
M2e MSLLTEVET: The Achilles Heel of influenza is bicistronic
A human to human influenza virion detected by Altermune
natural killer cell attacks infected cell
Total DNA Linker
Today
Model system with Ab to Phenylarsonate and Haemophilus influenza bacteria in rats.
gt-/- mice Building linker to hemagglutinin peptide from H3N2 Made M2e peptides
Protection from Hib Bacteremia by Fab-Ars Linker
100 75 50 25 0 200 100 30 10
Fab-Ars Dose
3 % P rotected: percentage of rats in each treatment group that had sterile blood cultures, i.e. <20 Hib CFU/ml.
11 12 13 14 15 16 17 18
Protection of Neonatal Rats From Hib Bacteremia by Treatment with Anti-Arsonate Antibodiesand [Fab Anti-HibPS-Arsonate] Linker*
Group Anti-Ars Fab41-Ars (mg) ( g) 1 - - >1 x10 6 , 5.0 x 10 5 Hib CFU/ml of Blood † 2 3 4 5 6 7 8 9 10 0.1 - - - - - - - - - 200 100 30 10 3 1 0.3 0.1 5.6 x 10 5 , 5 x 10 5 , 2.7 x 10 5 , 1.5 x 10 5 , 1.0 x 10 5 , 6.9 x 10 4 4.0 x 10 5 , 7.0 x 10 4 , 6.0 x 10 4 >1 x 10 6 , 2.5 x 10 5 , 1.7 x 10 5 , 1.0 x 10 5 1.0 x 10 5 , 1.0 x 10 5 , 4.0 x 10 4 , 2.0 x 10 4 1.6 x 10 5 , 9.0 x 10 4 , 6.0 x 10 4 , 1.0 x 10 4 >1 x 10 6 , 2.6 x 10 5 , 9.0 x 10 4 , 3.0 x 10 4 >1 x 10 6 , >1 x10 6 , 6.4 x 10 5 , 1.7 x 10 5 >1 x 10 6 , >1 x 10 6 , 7.0 x 10 4 , 4.0 x 10 4 >1 x 10 6 , 4.1 x 10 5 , 1.8 x 10 5 , 1.5 x 10 5 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 200 100 30 10 3 1 0.3 0.1 <20, <20, <20, <20, <20, <20, <20, <20, <20, <20, <20 1.4 x 10 3 , <20, <20, <20, <20, <20, <20, <20, <20, <20, <20 1.2 x 10 5 , 1.2 x 10 4 , 2.5 x 10 3 , 2.2 x 10 3 , 2.2 x 10 3 , <20, <20, <20, <20, <20, <20, 1.1 x 10 5 , 2.8 x 10 4 , 2.7 x 10 4 , 1.3 x 10 4 , 9.8 x 10 3 , 6.2 x 10 3 , 4.9 x 10 3 , 2.4 x 10 3 , 2.4 x 10 3 , <20, <20 >1 x 10 6 , 6.4 x 10 5 , 1.8 x 10 5 , 1.1 x 10 5 , 5.1 x 10 4 , 2.5 x 10 4 , 1.7 x 10 4 , 4.6 x 10 3 , 4.6 x 10 3 , 4.3 x 10 3 , 2.3 x 10 3 >1 x 10 6 , 2.0 x 10 5 , 1.9 x 10 5 , 1.3 x 10 5 , 2.8 x 10 4 , 2.5 x 10 4 , 2.4 x 10 4 , 8.7 x 10 3 , 7.5 x 10 3 , 1.9 x 10 3 >1 x 10 6 , 6.2 x 10 5 , 5.4 x 10 5 , 4.5 x 10 5 , 3.0 x 10 5 , 7.5 x 10 4 , 6.3 x 10 4 , 5.2 x 10 4 , 2.1 x 10 4 , 1.3 x 10 4 , 1.6 x 10 3 >1 x 10 6 , 4.0 x 10 5 , 3.2 x 10 5 , 2.1 x 10 5 , 2.1 x 10 5 , 2.0 x 10 5 , 2.0 x 10 5 , 1.4 x 10 5 , 1.1 x 10 5 , 1.0 x 10 5 , 1.4 x 10 4
Aptamer binds to 161 aa peptide from H3N2 hemagglutinin Ruth Arnon, JBC 279, 2004 AATTGAATAAGCTGGTATGTTGGTCACATAACGCCAACGCCAAAACTCTGAGTCGGGAAATCACTCCCAATTA OH O P O O O
13
O N 3
+
HO OH OH O OH OH O OH O OH O HO
10
OH O NHAc O CuSO4, sodium ascorbate NH O AATTGAATAAGCTGGTATGTTGGTCACATAACGCCAACGCCAAAACTCTGAGTCGGGAAATCACTCCCAATTA O OH O P O O O O N NH O AcHN O N N HO
14
OH O HO O HO HO O HO O HO HO OH
8.10E+08 7.10E+08 6.10E+08 5.10E+08 4.10E+08 3.10E+08 2.10E+08 1.10E+08 1.00E+07 Mean virus titers
Appendix A Reduction of infectivity in vitro of influenza virus on chicken cells by Altermune linker (apt-gal).
cntrl 4.63E+08 1uM apt 1.27E+08 0.2 uM apt 9.67E+07 1 uM apt-gal 6.13E+07 0.2 uM apt-gal 5.73E+07
Degradation of Lethal Factor Aptamer Cocktail or Protective Antigen Cocktail, Unmodified (UN) versus Modified (PEG or Phosphorothioate) Under Varying Conditions
CTRL 1h 2h 4h 6h CTRL 2h 4h 6h 24h
UN Th UN Th UN Th UN Th UN Th 70% FBS
CTRL 5 min. 15 min. 30 min.
UN PEG UN PEG UN PEG UN PEG Nuclease 70% FBS
CTRL 1h 4h 6h 24h
UN Th UN Th UN Th UN Th UN Th UN=Unmodified PEG=PEG-modified Th=Phosphorothioate modified Nuclease
Survival Curve of A/J Mice Immunized with Human Serum, Challenged with BAS and Treated with α-gal PAA-12 Aptamer and Doxycycline
•
A/J mice were immunized intraperitoneally with either 1X PBS or 1% human serum 1X per week for 5 weeks.
•
Mice were challenged intranasally with either 1X PBS or 1X10 6 BAS (Sterne strain).
•
Mice were treated with either PBS (with or without human serum) or PAA-12 aptamer drug ( 75 ug /dose with human serum) at 2 hours following challenge and every 24 hours thereafter for 10 days and either PBS or doxycycline (1.5 mg/kg – mouse dose) at 12 or 24 hours following challenge and for every 24 hours thereafter for 14 days:
•
Survival was monitored daily for 28 days.
The Future:
Priming the immune system for specific diversion by Altermune
A set of immunogens is chosen for:
• • • Convenient synthesis Safety The distinct spectrum of immune responses provoked
Various immunogens with their resultant immune responses
M13 Filamentous Phage
•
Collaborators
Dr. Alex Lucas, Childrens’ Hospital of Oakland Research Institute, Oakland, CA • Dr. Rick Lyons, University of New Mexico, Albuquerque • • • Dr. Ron Cook, Biosearch, Novato, CA Dr. Carol Cardona, UC Davis, CA Dr. Jeeva Vivikananda, Dr. Jonathan Kiel, Brooks Air Force Base, San Antonio, TX