>> GOOD AFTERNOON, EVERYONE. WELCOME TO THE WEDNESDAY AFTERNOON LECTURE FOR TODAY. AND WELCOME ALSO TO THE PEOPLE WHO ARE WATCHING ON THE WEB. WE HAVE A VERY INTERESTING PRESENTATION ABOUT A TOPIC THAT PRESENTS A CONSIDERABLE PUBLIC HEALTH CHALLENGE AND THAT IS THE GROWING PRESENCE OF ANTIBIOTIC RESISTANCE AND THAT'S TRUE FOR PARTICULARLY BACTERIAL INFECTIONS THAT WE USED TO HAVE A REALLY WONDERFUL PHARMACY WORTH OF ANTIBIOTICS THAT WOULD WORK IN ALMOST ANY KIND OF BACTERIAL INFECTION AND NOW AS YOU KNOW THE INCREASED INCIDENCE OF HIGHLY RESISTANT ORGANISMS WHETHER IT'S MRSA, OR 1 OF THESE REALLY NASTY GRAM NEGATIVES IS CAUSING ANYBODY WHO'S TAKING CARE OF SICK PATIENTS TO GROW INCREASINGLY ALARMED ABOUT THE PATH THAT WE'RE ON. SO WE DESPERATELY NEED NOW INVENTIONS AND NEW IDEAS ABOUT THOSE HIGHLY RESISTANT ORGANISMS. I THINK THERE WAS SOMEBODY 50 YEARS AGO THAT WE HAD PRETTY MUCH SOLVED INFECTIOUS DISEASE AND WE COULD MOVE ON TO OTHER THINGS BECAUSE WE HAD THAT UNDER CONTROL. WELL TIME HAS PROVED THAT TO BE THE MISSED MARK. WE HAVE NEWLY EMERGING VIRUSES RIGHT AND LEFT. LAST YEAR EBOLA THIS YEAR ZIKA AND THE THREAT AS INFLUENZA IS THE GREATEST OPPORTUNITY FOR A DISASTROUS WORLD WIDE PANDEMIC FOR WHICH WE ARE SOMEWHAT OVERDUE. ANOTHER 1918 EVENT CANNOT IN ANY WAY BE CONSIDERED OUT OF THE QUESTION, SO WE NEED NEW THERAPEUTICS FOR INFLUENZA AS WELL. TODAY'S SPEAKER IS IN FACT IN THE MIDDLE OF ALL THAT AND WE'LL BE TALKING ABOUT THE DEVELOPMENT OF THERAPEUTICS FOR BOTH VIRAL AND THERAPEUTIC INFECTIONS, AND DR. MAN-WAH TAN IS DIRECTOR AND SENIOR SCIENTIST AT INFECTIOUS DISEASES DEPARTMENT AT GENENTECH. HE MOVED TO STANFORD WHERE FOR 12 YEARS HE WAS IN THE DEPARTMENT OF GENETICS AND MICROBIOLOGY AND IMMUNOLOGY. AND THEN IN 2012, MOVED TO HIS CURRENT ROLE AT GENENTECH, WHERE HE'S INVOLVE ED IN A NUMBER OF PROJECTS THAT ARE USING INNOVATIVE STRATEGIES TO GO AFTER AND TREAT INFECTIOUS AGENTS OF MEDICAL IMPORTANCE. AND AS I SAID AT THE OUTSET, WE NEED THIS MORE CONSIDERING THE EPIDEMIOLOGY OF THESE DIRECTIONS THAT THE INFECTIONS SEEM TO BE GOING. SO IT'S A GREAT PLEASURE TO HAVE HIM HERE AS OUR SPEAKER. PLEASE JOIN ME IN WELCOMING DR. MAN-WAH TAN. [ APPLAUSE ] >> HI, GOOD AFTERNOON AND THANK YOU FOR THE OPPORTUNITY TO COME AND PRESENT TO YOU SOME OF THE SCIENCE THAT HAS BEEN GOING ON AT GENENTECH. SO THIS SLIDE IS ALSO FOR A DISCLAIMER, I WORK FOR GENENTECH, WHICH IS A MEMBER OF THE ROCHE GROUP, ALTHOUGH THE SIGN DOESN'T CHANGE WHETHER YOU DO IT IN THE COMPANY OR IN AN ACADEMIC SITUATION. SO TODAY I'M GOING TO TALK TO YOU ABOUT OUR APPROACHES IN USING MONOCLONAL ANTIBODIES AND INFECT YOWS DISEASES. I THINK MANGE OF US ARE VERY FAMILIAR WITH THIS KIND OF A GRAPH, SHOW WHAT,--WHAT I'M SHOWING HERE IS MORTALITY IN THOUSANDS PER YEAR FOR LUNG CANCER, STOMACH CANCER, COLORECTAL CANCER AND BREAST CANCER AND WE ALL KNOW HOW DEVASTATING THESE DISEASES ARE AND THE TOLL IT TAKES ON HUMAN CIVILIZATION, IN THE U.S., E. U. AND INDIA, MEXICO, BRAZIL, SOUTH CORIA AND TURKEY. YOU NOTICE THAD INFECT YOWS BACTERIAL INFECTIONS FOR EXAMPLE, IT'S 1.5 MILLION PEOPLE A YEAR. AND WHAT'S TROUBLING TO ME WHEN I LOOK AT A FIGURE LIKE THIS IS THAT THERE'S ENTIRE CONTINENT MISSING. WE DON'T EVEN HAVE AFRICA THERE. WE ONLY LOOKED FOR A COMPANY, IN PARTICULAR, WE ONLY LOOK AT AREA WHERE IS WE CAN DEVELOP DRUGS FOR WHICH THERE IS RETURN ON INVESTMENT, RIGHT? SO WE NOW LOOK AT CHRONIC HBV, AND LOOK AT INFLUENZA AND WHEN YOU LOOK AT THESE COUNTRIES IT IS COMPARABLE TO BREAST CANCER AND YOU LOOK AT BACTERIAL INFECTIONS IN THE U.S., AND YOU WILL FIND IT IS ALSO COMPARABLE TO BREAST CANCER FOR THESE COUNTRIES. AND ANOTHER TROUBLING FACT IS THAT DESPITE THE SERIOUS HUMAN AND ECONOM TOLL IT INFLICTS, INFECTIOUS DISEASES ONLY MAKE UP A FRACTION OF THE PUBLIC AND PRIVATE FACTORS AND GENENTECH ONLY ENTERED INTO THE INFECTIOUS DISEASE AREA ABOUT 9 YEARS AGO. AND SO WHAT CAN WE DO AND WHY DO WE DO INFECTIOUS DISEASE? IT IS BECAUSE OF THIS AS WELL THAT DR. COLLINS TALKED ABOUT AND I'M JUST USING BACTERIAL INFECTIONS AS AN EXAMPLE. SO YOU LOOK AT--THESE ARE THE CLASSES OF ANTIBIOTICS AGAINST BACTERIAL INFECTION. WHAT I'VE SHOWN HERE IS THAT THE LAST NEW CLASS OF ANTIBIOTIC WHICH WAS INTRODUCED TO THE CLINE NICK 2003, WAS ACTUALLY DISCOVERED IN 87. AND FOR EVERY SINGLE CLASS OF ANTIBIOTIC RESISTANCE HAVE EMERGED, USUALLY SHORTLY AFTER THE INTRODUCTION OF THESE ANTIBIOTIC. AND SO, AND WHAT I'VE SHOWN HERE ARE THE 2 OTHER IMPORTANT POINTS. MANY OF THE NEWER ANTIBIOTICS ARE INTRODUCED TO THE CLINIC ARE ACTIVE ONLY AGAINST GRAM POSITIVE BACTERIA: POINT NUMBER 2 IS IF YOU LOOK AT THE GREEN VERSUS BLACK, YOU WILL SEE THAT MAJORITY OF ANTIBIOTIC ARE ACTUALLY GIFTS FROM NATURE. THEY ARE NATURAL PRODUCT OR DERIVATIVES OF NATURAL PRODUCT. HAS NATURE ONLY GIVEN US NATURAL PRODUCTS? WE HAVE USED IMMUNE SERUM AGAINST TIP THERAPY ANDIA AND TETANUS IN 1890. SO WE THINK JUST AS WE UNDERSTAND ABOUT OUR IMMUNE SYSTEM, ANTIBODY IS ALSO A VERY IMPORTANT COMPONENT OF DEFENSE AND COULD POTENTIALLY BE HARNESSED FOR TREATMENT OF INFECTIOUS DISEASES. THAT'S WHAT NATURE DOES WITH ANTIBODY. AND ANOTHER POINT I WANT TO RAISE IS THAT IF YOU NOW LOOK AT GRAM NEGATIVE INFECTION, THE SITUATION IS EVEN MORE DIRE. THE LAST CLASS WAS GREWSED IN 1968 AND IT WAS DISCOVERED IN THIS 1961. IT ONLY TOOK 7 YEARS TO DEVELOP THE DRUG IN THE 70S AND 60S AND NOW IT TAKES 15-20 YEARS TO DEVELOP A NEW DRUG AGAINST NOW. SO THERE ARE SEVERAL CHALLENGES THAT PEOPLE WOWORK IN THE INFEBLGHTIOUS DISEASE AREA FACE. SO TODAY I WILL TELL YOU 2 STORIES ON HOW WHY CAN HARNESS THE OTHER GIFTS FROM NATURE, THE ANTIBODIES AND WE'RE TALKING ABOUT MONOCLONAL ANTIBODIES TO TREAT INFLUENZA A INFECTIONS AND THEN THIS NOVEL THERAPEUTIC PLATFORM THAT WE CALL ANTIBODY ANTICONJUGATE AND I WILL USE VIREMIA AS AN EXAMPLE BUT WE THINK IT COULD BE EXTEND TO THE S. AURUOUS AS WELL. SO WE WANT TO ACKNOWLEDGE THE PEOPLE WHO WORKING ON THIS PROJECT, THEY ARE THE 1S THAT NUCLEATED THE TEAM THAT BROUGHT ABOUT THIS MONOCLONAL ASPECT BODY. SO AS DR. COLLINS TALKED ABOUT EARLY ON, INFLUENZA INFECTION IS AN IMPORTANT DISEASE. HERE IS EPIDEMIOLOGY. OVER 600,000 HOSPITALIZATIONS IN THE U.S. AND E. U. THIS IS THE NUMBER FOR NONPANDEMIC SEASONINGS. AND 25,000 DEATHS IN U.S. ALONE. MANY OF US ARE VACCINATED WHICH IS GREAT AND I ENCOURAGE THAT STRONGLY BUT ONLY PARTIALLY EFFECTIVE: AFRICA REALLY DEPENDS ON IMMUNO COMPETENCE AS WELL AS ANTIGEN MATCH. SO AS I GET OLDER AND I KNOW THAT IS AN INEVITABLE FACT I KNOW MY IMMUNE COMPETENCY WILL REDUCE AND VACCINATION WILL NOT WORK AS WELL FOR ME AND MANY OF US IN THE FUTURE. AND IN 2014 FOR EXAMPLE, THE CDC SHOWED THAT VACKIZATION EFFECTIVENESS WAS ONLY 18% AGAINST H3 N2 BECAUSE OF THE MISMATCH. AND THE INFLUENZA VIRUSES, INFLUENZA A ACCOUNTS FOR 80% AND B ACCOUNTS FOR 20% AND THIS IS MORE DETAIL ON WHY WE FIND IT IS AN UNMET NEED. SO NO THERAPY DEMONSTRATED CLINICAL BENEFITS ON PATIENTS THAT ARE HOSPITALIZED WITH INFLUENZA, SO THAT IS THE UNMET NEED THAT WE'RE TRYING TO ADDRESS. THE STANDARD OF CARE RIGHT NOW IS SUPPORTIVE CARE AND NEUROINHIBITTOR. SO THE NEXT SLIDE I WILL TALK ABOUT A GENERAL, BRIEF INTRODUCTIONOT LIFE CYCLE OF INFLUENZA, SO INFLUENZA IN AN RNA VIRUS WITH 8 SEGMENTS HERE, IS SHOWN HERE, THERE ARE 3 IMPORTANT EXTRA CELLULAR COMPONENTS: OR CELL ASSOCIATED GLYCOPROTEINS, HEMOGLUTE9 AND H2 CHANNELS AND THESE ARE ALL TARGETS OF DRUGS. THE NEUROAMINIDACE VIRAL RELEASE, VEHICLE NATION BINDS TO THE HEAD REGION OF THE HEME LOW GLUTENIN AND IT'S THE ACID RECEPTOR. HA ALSO MEDIATES THE FUSION BETWEEN THE ENDOSOME AND THE HOST MEMBRANE, THE VIRAL MEMBRANE AND THE HOST MEMBRANE TO RELEASE VIRAL CONTENT FOR TRANSCRIPTION AND TRANSLATION AND THEN THE NEUROMITT DACE RELEASES THE BUDDING VIRUS TO CAUSE NEW INFECTION. SO NEUROMINIDACE INHIBITORS, AND TAM OVIR BLOCKS THIS PROCESS AND MANY OF THE GLOBULINS BLOCK THIS ATTACHMENT. HOWEVER, WHEN WE STARTED TO LOOK AT THIS PROBLEM, WE REALIZE THAT THE REASON WHY WE NEED TO HAVE DIFFERENT VACCINES EVERY YEAR IS BECAUSE THESE--THE 2 MAJOR CIRCULATING HA VIRUS IN THE HUMAN ARE THE H1 AND H3 AND WHEN YOU LOOK AT HEMOGLUTENIN SEQUENCE, THEY'RE ONLY 48% IDENTICAL ESPECIALLY WHEN YOU LOOK NOW AT THE HEAD REGION. SO THIS IS A COMPILATION OF OVER 11,000 HA FROM THE 16 SUBTYPES OF H. A. THAT EXIST IN NATURE. SO THEY FALL INTO 2 GROUPS. AND SO WE HAVE TO BE VACCINATED WITH THE H. A. CORRESPONDING TO GROUP 1 AS WELL AS GROUP 2. THERE ARE HOWEVER, A SMALL MINORITY OF ANTIBODIES THAT'S BEEN PRODUCED BY SOME OF US. THEY ARE ACTUALLY BINDING TO A CONSERVED REGION AND WE LEARNED OF THE EXISTENCE THAT REPORTED THAT WHEN A CLONE OF A HELPED THOUSAND B-CELLS THEY FOUND 1 BROADLY NEUTRALIZING ANTIBODY. SO THERE IS THAT NEEDLE IN THE HAY STACK THAT IS WORTH LOOKING FOR BUT IT IS A TINY LITTLE NEEDLE IN A VERY, VERY BIG HAY STACK. SO CAN WE LOOK FOR THESE BROADLY NEUTRALIZING ANTIBODY? WHAT WE WANT IS 1 THAT BINDS AND TARGETS BOTH GROUP 1 AND GROUP 2 SO THAT IT WILL BE ABLE TO NEUTRALIZE ALL OF THEM AND THAT IT IS MORE EFFICACIOUS THAN THE STANDARD OF CARE. SO IT IS WITH THAT GOLDEN MIND THAT WE DEVELOP, WE DEVELOP A STRATEGY TO FIND THAT NEEDLE IN AT THIS TIME B-CELL HAY STACK. SO THE APPROACH IS AS FOLLOWS. INITIALLY, WE GET PBMC FROM VACCINATED DONORS AND VACCINATED WITH HA 3, HA-1 FROM THESE 2 STRAINS AND PBMC ARE THEN ACTIVATED INVITRO USING VERY DIFFERENT HA SUBTYPES. THESE PBMC ARE INJECTED INTO THE SPLEEN OF KID MICE AND THEN AFTER 8 DAYS, THE SPLEENIC PLASMA BLASTS ARE BEING--THE EFFECTS ARE SORTED USING AGAIN A DIFFERENT SUBTYPE AND THEN THE PROFILE OF THIS ANTIBODY IS THEN TESTED FOR THE ABILITY TO BIND ACROSS OTHER SUBTYPES. SO IT IS WITH THIS WAVE OF SELECTING FOR VIRUSES THAT WE WERE ABLE TO CLONE 84--840 HUMAN MAPS, 20 OF THESE BIOMULTIPLE SUBTYPES BUT WE WERE EXCITED TO FIND THAT 2 OF THESE ANTIBODIES BIND AND NEUTRALIZE BOTH GROUP 1 AND GROUP 2. H. A., 1 IS SPECIFIC TO GROUP 1 AND ANOTHER SPECIFIC TO GROUP 2. WE THOUGHT MAYBE WE WERE LUCKY BUT WE HAVE ACTUALLY DONE THIS EXPERIMENT NOW AND FOUND ANOTHER BROADLY NEUTRALIZING ANTIBODY IN GROUP B BUT I WON'T TALK ABOUT THAT BECAUSE THIS SCHEME IS SIMILAR. SO WHAT DOES THIS ANTIBODY DO? SO WHEN WE LOOK FOR THE ABILITY TO NEUTRALIZE HIRKS 1, 2, AND 3, THE OTHER 1 VS BEEN SHOWN TO BE INFECTING HUMANS AND SOME OF THESE ARE CAUSING PANDEMIC OUTBREAK, CAN YOU SEE THEY ARE ABLE TO NEUTRALIZE ALL OF THESE STRAIN AND WE HAVE ALSO TESTED IT AGAINST OTHERS. SO THESE ARE 1S THAT OCCASIONALLY INFECT HUMAN LIKE H5 OR H7 AND YOU CAN SEE THAT THEY ALSO NEUTRALIZE H6, H7, AND OTHERS TYPES THAT IT COULD OCCASIONALLY INFECT HUMAN AND POTENTIALLY BE THE NEXT PANDEMIC AS WELL. SO HOW DOES THIS ANTIBODY NEUTRALIZE THE VIRUSES. SO THE MAIN ANTIBODY, IGG 1 WE PRODUCE THAT VAC NATION BLOCKS, HA BY SIMPLY BINDING TO THE HA AND PREVENTS HA FROM BINDING TO THE ASILIC ACID ON THE BLOOD CELLS, SO WHEN IT BINDS TO A RED BLOOD CELL WILL CAUSE THIS FORMATION OF LATTICE AND WILL THEN--SO VIRUSES WILL FORM FROM THIS LADDER AND THEN THE ANTIBODY THAT PREVENTS THE BINDING WILL THEN HAVE THE HEMOGLUTENATION AS SHOWN HERE. A HEAD ANTIBODY THAT PREVENTS THIS BODY WHERE YOU HAVE THE SEDIMENTATION OF THE RED BLOOD CELL. SO THE ANTIBODY THAT WE FOUND IN WHICH WE CALL 4549A BECAUSE IT'S IN CLINICAL DEVELOPMENT SO WE GIVE IT A MORE EFICIAL NAME HAS THIS HEMOGLUTENATION PHENOTYPE. SO THE VIRAL BINDING BLOCKS IT FROM THEN PREVENTS THE HEMOGLUTENATION. AND I TOLD YOU ANOTHER FUNCTION IS TO ALLOW FOR FUSION BETWEEN THE VIRAL MEMBRANE AND THE VIRAL AND THE HOST MEMBRANE SO WE USE THIS FUSION ASSAY WHERE H. A. HAS BEEN EXPRESSED IN THE CELLS, AND THEN AT THE Ph DROPS, CHAI IS WHEN WE UNDERGO CONFIRMATIONAL CHANGE AND WE'RE UP FOR NEUTRALIZATION OR THE Ph ASK WHY THAT IS BEING FORMED. SO IN HEAD BINDING ANTIBODY DOES NOT PREVENT THE CONFIRMATIONAL CHANGE AND YOU SEE THE [INDISCERNIBLE] WHERE THE ANTIA AND ANTIBODY PREVENTS THIS FORMATION AND SO, MORE IMPORTANTLY, WE FOUND THAT THIS ANTIBODY IN FACT ALSO BINDS AND REMAINS BOUND TO THE HA EVEN AT ACIDIC Ph. JUST 1 FUNCTION OF THE AGENT BODY FROM THE FAB REGION OF ANTIBODY BUT ANTIBODY WE KNOW ESPECIALLY IN CONTEXT OF IGG1 WHICH IS WHAT THE ANTIBODY IS, HAS AN SC FUNCTION. AND 1 OF THE FUNCTIONS OF SC IS TO RECRUIT SC GAMMA CONTAINING CELLS AS SHOWN HERE, SO HAVE YOU HERE--AS VIRUS INFECTED CELLS SHOW BUDDING OF VIRUSES SO NOW YOU HAVE THE DISPLAY OF HA OR NA OF THE DISPLAY OF THE CELLS, SO THE ANTIBODY WILL BYPASSED TO THE HA IN THIS CASE AND THEN AT THE REGION WILL RECRUIT THE FC GAMMA RECEPTORS AND THEN THE NK SELLS WILL PRODUCE AN INTIME THAT WILL KILL THE INFECTED CELLS. SOPHISTICATEDY WE ASK THE QUESTION WHETHER THE ANTIBODY CELL TRIGGERS THE ANTIBODY CELLULAR CYTOTOXICITY. SO TO ASK THAT QUESTION WE OBTAIN BLOOD FROM VOLUNTEER DONORS AND NOW ASK THAT INFECTED CELLS IN THE PRESENCE OF INFECTED CELLS WHETHER NK CELLS ARE ACTIVATED. SO THIS IS OUR ANTIBODY, YOU CAN SEE THAT THE PERCENT LAMP 1 STAINING AND THE RITUXMACK, WHICH IS AN ANTIBODY, COME IS ABSENT IN THE A519 CELLS DO NOT TRIGGER THE ACTIVATION OF THE NK CELLS AND THEN WE ASK ALSO IF IT THE INFECTED CELLS ARE BEING KILLED BY MEASURING THE LDH ABSORB ANT AND YOU CAN SEE HERE THAT THE CELLS ARE INDEED DEAD--THE INFECTED CELLS ARE BEING KILLED IN THE PRESENCE OF ANTIBODY. SO WE HAVE 2 MECHANISMS OF ACTION, 1 BY BINDING TO THE HA, FUSION AND THE OTHER IS TO RECRUIT IMMUNE CELLS TO KILL INFECTED CELLS. SO WHERE EXACTLY DOES THE ANTIBODY BIND? SO WE CRYSTALLIZE THE HA, IT'S A TRIMER WITH THE FAB OF OUR ANTIBODY AND INDEED IT BINDS TO THE CONSERVED REGION, THE LIGHT CHAIN BINDING SHOWN HERE AND THE HEAVY CHAIN SHOWN ON THE RIGHT. AND WE HAVE ANALYZED THE AMINO ACIDS. THERE ARE ABOUT 32 CONTACT POINTS BUT IT'S ANTIBODY AND THEY'RE CONSERVED ACROSS INDEED ALL THE HA TYPE THAT I MENTIONED EARLIER ON. WE WONDER THEN WHETHER RESISTANCE COULD EMERGE VERY EASILY AND THAT'S ALWAYS THE QUESTION THAT BACKS AND OFTEN TRIPS THE DISCOVERY AND EFFECTIVENESS OF DRUGS. SO WE'VE DONE PASSAGING OF THE VIRUSES AND INCREASING CONCENTRATION OF THE ANTIBODY. OVER WEEKS. WE WERE NOT ABLE TO FIND ANY RESISTANT MUTE ANT, USED THE WAY WE PASS SH1. WE DID FIND 3 ALLELES, 1 IN THIS RESIDUE AND ANOTHER 2 IN THE 391 RESIDUE. SO NOW WE'RE DESCRIBING THE RESISTANCE HERE, 387 K PREVENTS BINDING OF THE ANTIBODY. THESE 2 ALLELES DOES NOT PREVENT BINDING OF THE ANTIBODY AT NEUTRAL Ph BUT DOES IMPACT SLIGHTLY THE BINDING ACIDIC Ph AND ACTUALLY FUSES AT THE HIGHER Ph THAN WILD-TYPE VIRUS. AND WE THINK THAT ALTHOUGH WE HAVEN'T GENERATED THE DATA IN ANIMALS THAT WE ARE--WE SHOULD BE ABLE TO DEAL WITH THIS MUTATIONS BECAUSE ANTIBODY STILL BINDS TO HA AND INFECTED CELLS AND SHOULD RECRUIT IMMUNE CELLS TO KILL INFECTED CELLS. WE IN FACT HAVE THAT DATA FOR INFLUENZA B WHICH SHOULD THAT WE DON'T HAVE THE INFLUENZA A. SO ANOTHER IMPORTANT FACTOR HERE IS THAT ALL 3 ALLELES ARE LESS FIT IN OUR INVITRO STUDIES. SO WE DON'T KNOW THE CONTRIBUTION OF THESE ALLELES IN HUMAN, IN INFECTION, GIVEN THAT THIS MOLECULE IS ALREADY IN CLINICAL TRIAL. THIS IS--THESE ALLELES ARE THE 1S WE ARE MONITORING IN PATIENTS. SO THIS IS IN A CARTOON FORM, HOW THE ANTIBODY WORKS, SO SHOWN HERE IT'S A HEAVY CHAIN AND LIGHT CHAIN BOUND TO THE CONSERVED REGION OF THE HEMOGLUTENIN AND SO THIS IS JUST NORMAL PART OF THE TRIMER. YOU SEE HERE THIS IS THE FUSION PEPTIDE, AND SO AS THE Ph DROPS, DIFFUSION PEPTIDE AND THE HA HERE HELIX, AV HERE THEN FORMS A HELIX TO EXTRUDE THE FUSION PEPTIDE SO THEN IT BINDS TO THE WHOLE CELL AND THAT BRINGS THE VIRAL MEMBRANE AND HOST MEMBRANE TOGETHER TO ALLOW FOR THE RELEASE OF VIRAL GENOME. AND THE MHCA 549 A BINDS TO THIS REGION AND PREVENTS THE CONFORMATIONAL CHANGE AND THAT THE MECHANISM THAT PREVENTS THE VIRAL FUSION. SO IT WORKS WELL IN VITRO, THE QUESTION IS HOW WELL DOES IT WORK IN VIVO. SO WE HAVE 2 ANIMAL MODELS. ONE IS THE MOUSE MODEL, USING THE DDHA, 2 J STRAINS A GOAL OF THIS PROJECT IS TO TREAT PATIENT WHO IS ARE--WHO HAVE SEVERE INFLUENZA. SO THE CHALLENGE HERE IS TO TREAT MICE LATER AFTER INFECTION. SO HERE WE CHOSE A TIME, 72 HOURS POST INFECTION AT OUR CUT OFF AND ASK WHETHER THIS ANTIBODY WORKS AND YOU CAN SEE FOR GROUP 1 AS WELL AS IN GROUP 2, WE SEE A DOSE DEPENDENT PROTECTION IN THIS SEVERE MODEL. SIMILARLY IN FERRET, YOU CAN SEE HERE THAT IF YOU INITIATE TREATMENT 72 HOURS POST INFECTION, ALSO TAMIVIR DOESN'T WORK VERY WELL AND THAT SEEMS TO ALSO SHOW AND BE TRUE IN PATIENTS. MHC-5 494 A IS STILL PROTECTED. AND EVEN IN SITUATIONS WHERE PATIENTS OR IN THIS CASE, THE MOUSE MODEL IS BEING TREATED WITH MHAA AT SUBEFFICACIOUS DOSE BUT IF THEY ARE USED IN COMBINATION, YOU SEE HERE THAT THEY ARE ACTUALLY PROTECTIVE. SO, THAT MAKES SENSE GIB THAT THE MECHANISM OF ACTION OF THESE DRUGS ARE DIFFERENT. SO THE CLINICAL TRIAL IS REALLY TO HAVE PATIENTS WHO ARE ALREADY REQUIRESOX GENERATEDDATION WHO ARE ON TAMI FLU AND ON TOP OF THIS WE ADD MHAA 5449 A. SO WE DON'T KNOW THE RESULT OF THIS TRIAL BUT WHAT I HAVE TOLD YOU TODAY IS THAT WE HAVE AN ANTIGEN SPECIFIC IN VIVO PLASMA BLAST. THEY'RE EFFICIENTLY IDENTIFIED, VERY RARE FUNCTIONAL ANTIBODY. THIS ANTIBODY IS BROADLY NEUTRALIZING AND IT HAS 2 MECHANISMS OF ACTION, IT'S BLOCK M MEDIATED MEMBRANE FUSION, IT ALSO TRIGGERS ADCC. WE HAVE SHOWN THAT THESE ANTIBODY NEUTRALIZES ALL SEASONAL AND PANDEMIC STRAINS OF INFLUENZA AND THAT THEY SYNERGIZE WITH A TAMI FLEW INFLUENZA TREATMENT OF INFECTION IN THIS ANIMAL MODEL. WE HAVE ALSO DONE A HUMAN CHALLENGE STUDY WHERE WE SHOWED THAT THESE ANTIBODY IS INDEED WELL TOLERATED WITH GOOD SAFETY PROFILES, LOW IMMUNO GENERATEDISSITY RATE, HAS A PSEUDOPK THAT IS CONSISTENT WITH IGG1 WHICH IS 21 DAYS OF HALF LIFE AND IN THE CHALLENGE STUDY WE SHOW THAT THE INFLUENZA LOADS AND SYMPTOM SCORES WERE SIPPING 95 CANTILY DECREASED COMPARED TO PLACENTA SEEK SEEK O AT A DOSE OF 3600-MILLIGRAMS. SO HOPEFULLY IN A FEW YEARS WE WILL BE ABLE TO SEE HOW TRULY THIS ANTIBODY PERFORMS IN SEVERELY ILL PATIENTS IN OUR GLOBAL TRIAL FOR INFLUENZA. I WILL NOW SWITCH GEARS AND TALK ABOUT ANOTHER UTILITY OF THE ANTIBODY AND THAT IS THE PLATFORM USING ANTIBODY THAT IS CONJUGATED TO AN ANTIBIOTIC USING A CLEAVABLE LINKER AND I WILL TELL YOU A STORY OF HOW THAT'S BEEN DEVELOPED TO TREAT STAFF AU REUS, ON THIS. AND I WILL SHOW THAT WHO WORKED ON THE PROGRAM, THE SMALL MOLECULE AND THEN DICK VANDLEN ON LINKING THOSE 2 MOLECULES TOGETHER. I THINK MANY OF US KNOW THAT STAFF AUREUS IS A ORGANISM THAT CAN CAUSE A VARIETY OF INFECTIONS IT RANGES FROM WOUND AND SOFT TISSUE INFECTIONS TO LIFE THREATENING ENDOCARTITEIS OR NECK ROUGH ATOMITIZING PNEUMONIA AND SEPTIC SHOCK. AND EVEN WITH WITH THE ADVANCEMENT OF THIS NEW ANTIBIOTIC I TALKED TO YOU ABOUT, LINEZOLID, AND DAPTOMYCIN,EE WE ARE SEEING THESE ASSOCIATE WIDE THE INFECTION AND BETWEEN 10-20,000 PATIENTS DIE OF MRSA INFECTION EACH YEAR. SO THE QUESTION THAT CAME TO MIND WAS WHY DO ANTIBIOTIC THERAPY FAIL EVEN WITH THE NEW CLASS OF ANTIBIOTIC. SO TO ADDRESS THAT QUESTION ACCIDENT WE WENT AND LOOKED AT THE INFECTION CYCLE OF STAFF AUREUS. SO JUST TO RECAP WHAT I TOLD YOU HERE, DESPITE APPROPRIATE ANTIBIOTIC TREATMENT, 40% OF PATIENTS HAVE PERSISTENT BACTERIAEREMMIA, IT'S MORE THAN 4 DAYS AND 70% OF THE PATIENTS WHO HAVE PERRIST SENT DAN DEVELOPED THE METASTATIC TREAD AND THAT LEADS TO 20% MORTALITY. SO WHAT MAY BE GOING ON HERE SO TYPICALLY STAPH AUREUS IS ENTER THROUGH AN OPEN WOUND AND WHEN IT ENTERS THE BLOOD STREAM IT'S RAPIDLY INTERNALIZED BY NUTRIFILLS. SO AND NUTRIFILLS IS FIRST LINE OF DEFENSE. SO WESTBOUND THESE CELLS, STAPH AUREUS CAN SURE VIVE AND IS NOT ONLY SURVIVES BUT IT USES THIS SPREAD FLEW THE BLOOD STREAM TO THE OTHER TISSUES, THE BONE, LUNG, HARD VALVE AND THE LUNG. AND THEN, I WILL LYSE NUTRIFILLS AFTER SEVERAL DAYS. STAPH IS ALSO ENDOW WIDE A VARIETY OF PROTEINS ON THE SURFACE TO INNOVATE NONPHAGOCYTIC CELLS. SO IT USES NONBINDING PROTEIN SUCH AS A AND B THAT BINDS TO FIBER LECTIN AS WELL AS NMD, DRKSA AND INSIDE THE CELLS STAPH PROLIFERATES AT SMALL COLONY VARIANT AND CAN PERSIST FROM DAYS TO WEEKS. OVER THE PROCESS OF IT BEING IN THE CELLS, IT UNDERGOES TRANSCRIPTIONAL CHANGE AND IT STEADILY INCREASES THE LEVEL OF THE BINDING PROTEIN SO THAT WHEN IT LYSIS, THE EPITHELIAL CELLS AND THE NONPHAGOCYTIC CELLS IT'S TRYING TO REINVADE THE OUTER CELLS. SO STAPH CAN INVADE THE PHAGOCYTIC CELLS AND CAN SO WE ASK WHAT THE RESDENSE OF THE THIS ON THE CELLS ON THE EFFICACY OF THE ANTIBIOTIC. AND SO THE QUESTION WE ASK IS WHAT IS THE MINIMUM RECONCENTRATION OF ANTIBIOTIC NEEDED TO KILL STAPH AUREUS, WHEN THEY'RE GROWING CELLULARLY AND THIS IS THE TYPICAL MICRO BIOLOGY ASSAY WHY DO HERE IN A MICRO BIOLOGY LAB. AND YOU CAN SEE IT'S 4 NANO GRAMS PER MILL. HOWEVER WHEN STAPH AND INSIDE THE CELL AS SHOWN HERE THAT THEY'RE NOW WITHIN MACROPHAGE, UP SEE HERE THAT THE CONCENTRATION NEEDED TO KILL THE INTRACELLULAR BACTERIA IN FACT SIGNIFICANTLY INCREASED HIGHER THAN IT COULD BE ACHIEVED SO THE C-MAX OF THE SERUM IS LOWER THAN THE CONCENTRATION NEEDED TO KILL EPITHELIAL RACELLULAR BACTERIA. SO WE HYPOTHESIZE THAT INTRACELLULAR BACTERIA ARE REFRACTORY TO ANTIBIOTIC AND THIS IS THE REASON FOR THERAPEUTIC FAILURES USING ANTIBIOTIC. SO WE WANT TO DEVELOP A FEW ASSAYS TO SEE IF WE CAN RECAPITULATE THIS HYPOTHESIS. AND SO HERE WE USE A FEW HUMAN CELL TYPES. A549 IS LOW CELLS, HUMAN BLOOD AND MICRO EPITHELIAL CELLS. YOU SEE HERE THAT STAPH AUREUS, CAN GROW VERY WELL IN THOSE CELLS, AND THEN PLANK TONIC CELLS CAN BE KILLED BY SANK O MICEIN AND HOWEVER, WHEN THIS SEALS ARE FIRST INFECTED WITH STAPH AUREUS, SO STAFF AUREUS IS NOW EPITHELIAL RACELLULAR, SANK O MICEIN IS NO LONGER ABLE TO KILL THIS BACTERIA. THAT'S TRUE FOR EPITHELIAL CELLS, ENDOTHELIAL CELLS AND OSTEOBLAST. SO THAT'S HUMAN CELLS. WHAT ABOUT IN THE MOUSE INFECTION MODSLE?--MODEL? THIS IS A TYPICAL INFECTION MODEL WE USE TO DETERMINE THE EFFICACY OF AN ANTIBIOTIC. SO BACTERIA MATERIAL HAS GROWN AS PLAN TONIC STAGE USED TO INFECT MICE THAT'S BEEN TREATED WITH YOUR PDF CONTROL OR WHATEVER SOLVING CONTROL AND THEN COMPARED TO A MOUSE THAT IS BEING TREATED WITH SANK O MICEIN, AND YOU CAN SEE HERE IN THIS EXPERIMENT, SANK O MICEIN IS VERY EFFECTIVE AT CLEARING INFECTION. HOWEVER, IF YOU ADD A WRINKLE TO THIS EXPERIMENT BY FIRST RECOVER BACTERIA SO YOU DO THIS EXPERIMENT HERE, BUT YOU NOW RECOVER BACTERIA THAT ARE ALREADY INFECT THE CELL BY TAKING PERO TON EEL LAVAGE AND NOW USE THIS CELL TO INFECT MOUSE IN THE PRESENCE OR ABSENCE OF VANCO MICEIN, EVEN WITH VANCO PRESENT, THE KIDNEY OF THESE MICE ARE STILL--COULD BE INFECTED AND THERE IS PROLIFERATION. AND SO THAT SUGGESTS TO US IS THAT ABLATING INTRACELLULAR STAPH AUREUS IS KEY TO CLINICAL SUCCESS. SO WITH THAT HYPOTHESIS IN MIND, WE WANT TO TEST WHETHER THE HYPOTHESIS HOLDS TRUE AND WHETHER WE CAN DEVISE THIS STRATEGY TO KILL THIS CELLULAR BACTERIA. SO THAT LED US TO THINK ABOUT THE ANTIBODY, ANTIBIOTIC CONJUGATE WHERE OUR IDEA HERE IS TO OPTIMIZE, OR COAT STAPH AUREUS, AND THEN WE HAVE A LINKER, THAT HAS AN ANTIBIOTIC. SO WHAT WILL HAPPEN THEN IS THAT THIS AAC WHICH I CALL IT NOW, WHILE TECHNICALLY IT IS A BIOMAP, ANTIBI BODY CONJUGATE BECAUSE WE HAVE ENGINEERED THIS IN THE LYSEEN CHANGE SO THIS CARRIES TOO ANTIBIOTICS PER MOLECULE. SO THIS IS REALLY WHAT THE MOLECULE LOOKS LIKE. SO HAVE YOU BACTERIAL THAT IS IN THE BLOOD STREAM THAT WILL BE COATED WITH THE A. A. C. AND THEN IT'S OPTIMIZATION WILL CAUSE THE BACTERIAL TO ENTER THE CELLS IN MACROPHAGE, NUTRIFILLS, OR GAMMA CONTAINING CELLS OR KILL THE BACTERIA. OR STAPH AUREUS, FROM THE PHAGOCYTIC CELLS WILL VERY QUICKLY BE CAPTURED BY THE AAC AND DRIVE IT INTO FC CONTAINING CELLS FOR KILLING OR FOR CELLS THAT CONTAIN--EPITHELIAL CELLS THAT CONTAIN THE STAPH AUREUS CONTAINING, BECAUSE STAPH WANTS TO IN THE ENDOTHELIAL CELLS, SO IF THEY ARE ALREADY BEEN COATED WITH THE AAC, AS A ENTER THE CELLS, THEY ALSO BRING THE AAC INTO THE INFECTED CELLS, SO UNDER THE SUPER INFECTION CONDITION, YOU ARE ALSO BRINGING IN THE DRUG TO PROVIDE THIS BI STANDARD KILLING. SO, I WANT TO SHOW YOU WHAT HAPPENS AND I WILL PRESENT YOU THE DATA TO SUPPORT WHAT HAPPENS. SO WHAT HAPPENS HERE IS THAT WITHIN THE PH AGOLYSOSOME, THIS VALVE CLEAVER IS ENGINEERED BY CLEAVE, WE ENGINEERED IT TO BE CLEAVED BY [INDISCERNIBLE] AND SO WE HAVE ENGINEERED A PATH HERE THAT IS SELF-EMULATING AND SO IT WILL THEN DESTROY ITSELF AND NOW RELEASE THE ACCESS AND FREE ANTIBIOTIC INSIDE THIS VERY CONFINED ENVIRONMENT IN CLOSE PROXIMITY TO THE BACTERIA. AND THAT'S HOW WE ARE ABLE TO BRING HIGH CONCENTRATION OF ANTIBIOTIC TO THE BUG. SO WHAT IS NEEDED IN ORDER FOR THIS TO WORK? SO WE NEED AN ANTIBIOTIC THAT HAS HIGH POTENCY, ACTIVE IN LOW Ph BECAUSE THIS IS THE ENVIRONMENT THAT THE ACCESS OF THE ANTIBIOTIC IS RELEASED AND IT NEEDS TO BE CAPABLE OF CONJUGATION. SO WE HAVE TRIED MANY DIFFERENT ANTIBIOTICS AND THE 1 I WILL TALK TO YOU ABOUT IS A DERIVATIVE OF RIFASMIN, YOU CAN SEE THAT THE PH 5 AND 7 ARE EQUIVALENT. THE AAC IS COMPARABLE, MAYBE I LITTLE LESS EFFICACIOUS, AT Ph 5 AND 7. BOTH COULD BE RELEASED BY CITAFSIN, HOWEVER, RIFAMPIN IS NOT FOR THE AAC AND THE REASON YOU CAN SHOW IN THIS GRAPH IS THAT IT HAS A SHORT RETENTION TIME WITHIN THE CELLS. SO THESE CELLS ALONG WITH THE FAG O SIGNIFYITOSE HERE, AND YOU CAN SEE HERE BY MOST OF THIS BY RIFAMPIN IS RELEASED IS NOW BEING FOUND INSIDE OF THE CELL WHEREAS YOU CAN SEE HERE THE RIFALOG CONTINUES TO BE RETAINED IN THE CELLS. SO 1 IMPORTANT FEATURE OF A POTENT ANTIBI THETIC IS WE WANT TO RELEASE AN ANTIBIOTIC INSIDE THE CELL AND IT'S RETAINED IN THE CELL. SO THIS, ANOTHER VERY IMPORTANT FEATURE OF THIS IS THAT NOW WE CAN USE A BROAD SPECTRUM ANTIBIOTIC AND KILLS BANTHERRIA AND BECAUSE THIS IS ONLY RELEASED INSIDE THE CELL. SO BY THE TIME YOU GET OUT OF THE CELL IT'S UNDERGONE ORDERS THAT LEAD TO DILUTION SO NOT ONLY DOES THE ANTIBODY PROVIDE SPECIFICITY ON THE DRUG AND WE CAN NOW BRING IN A BROADLY ACTIVE ANTIBIOTIC TO KILL A BUG AND IT'S UNLIKELY THAT THAT CONCENTRATION AFTER DILUTION WILL HAVE ANY IMPACT ON THE GUT MICRO BIOTA. SO THE QUESTION IS WHAT IS IT? WHAT ADDITIONAL FEATURES MAKE THIS RIFALOG MORE ACTIVE? AND SO HERE WE WERE LUCKY. SO IN ORDER FOR US TO LINK THIS MOLECULE. WE NEED MODIFICATIONS, MADE MOLLIFICATIONSOT LEFT-HAND SIDE AND NOW WE SEE THAT RIFA LOG GAIN ACTIVITY THAT WAS PREVIOUSLY ABSENT IN THE RIFAMPIN. SO WE DID THE ABILITY TO KILL NONREPLICATING BACTERIA AND IT'S STATIONARY FAITH ARE PUT IN PBS AND YOU CAN SEE HERE OVER TIME THAT BACTERIA DON'T SURVIVE THAT WELL IN PBS AND THERE'S A LOG REDUCTION, BUT CAN YOU SEE HERE THAT RIFAMPIN IS NOT VERY EFFECTIVE AT KILLING STATIONARY PHASE BACTERIA BUT THE RIFA LOG IS ABLE TO DO SO. ANOTHER FEATURE WE BI BEGIN TO UNDERSTAND NOW FROM THE SCIENTIFIC COMMUNITY IS THAT WHEN BACTERIA ARE INTRACELLULAR THEY ENTER A STATE CALLED PERSISTERS. AND PERSISTERS ARE DEFINED BY CELLS THAT WHEN THEY'RE TREATED WITH ANTIBIOTIC ARE STILL ABLE TO SURVIVE. SO SAY CIPRO, YOU TREAT ANTI--BACTERIA WITH CIPRO AND IT'S ABLE TO KILL ABOUT 99.9% OF THE BACTERIA AND PICTURES 1% OF THE BACTERIA ALTHOUGH THEY ARE RESISTANT TO CIPRO, DO NOT HAVE GENETIC ALTERATION, THEY'RE SIMPLY A PHENOTYPIC MANNESTATION BECAUSE WHEN YOU RECULTURE THIS BACTERIA AND TREAT THEM WITH CIPRO, 99.9% OF THESE BACTERIA WILL ALSO BE KILLED AND THE .1% WILL SURVIVE AS PERSISTERS AND THOSE ARE INTRACELLULARLY DISTINCT. SO WE ASK THE QUESTION, WHETHER THE RIFA LOG OR RIFAMPIN HAD ANY IMPACT ON THE PERSISTER CELL. SO WE ADDED A SECOND ANTIBIOTIC. SO BY ADDING RIFAMPIN IT DOESN'T DO ANYTHING OF THE ABILITY TO KILL THE INTERRACELLULAR PERSISTERS HOWEVER RIFALOG CAN ELIMINATE TO THE LIMIT OF DETECTION THE PERSISTERS AS WELL. SO WE HAVE FORTUNATELY GAINED 2 M. O. A. WE DON'T KNOW IF THEY'RE DIRECTLY RELATED OR NOT BUT WE KNOW IT'S ABLE TO KILL NONREPLICATING BACTERIA AS WELL AS NONANTIBIOTIC RESISTANCE. SO WE HAVE THE ANTIBIOTIC PORTION OF THIS MOLECULE. SO NEXT WE NEED TO FIND THE ANTIBODY. SO INITIALLY WHEN WE WERE WORKING ON STAPH AND LEARNING FROM OUR INFLUENZA PROGRAM, WE THOUGHT THAT MAYBE WE CAN FIND NEUTRALIZING ANTIBODY THAT CAN KILL STAFF AUREUS, WE WERE LUKEY WITH INFLUENZA, WE LOOKED AND LOOKED VERY, VERY HARD FOR ANTIBODY THAT ARE ABLE TO BIND AND KILL STAPH AUREUS AND WE LOOKED AT OVER 40 MOAN O CLONAL ANTIBODIES AND WE TESTED COMBINATIONS UP TO 4 DIFFERENT ANTIBODIES AND NONE OF THESE WORKED. SO ANTIBODY ALONE FOR BACTERIAL INFECTION DOESN'T SEEM TO BE SUFFICIENT. SO THE NEXT QUESTION WE ASKED IS THAT CAN WE FIND ANTIBODY THAT BINDS HIGHLY ABUNDANT, HIGHLY CONSERVED, AND STABLY EXPRESSED EPITOPES. THAT MEANS THAT THEY'RE FATALLY EXPRESSED INVITRO AS WELL AS INFECTION AND THIS ANTIBODY AS RECOVERED FROM CONVALESCENT PATIENT AND THIS ANTIBODY FALLS INTO DIFFERENT CLASSES, SOME ANTIWIDES BINDS TO PROTEINS SO CAN YOU SEE THAT WITH THIS 1 THERE'S GOOD BINDING BUT INFECTION, THE CLUMPING FACTORA IS NOT WELL EXPRESSED SO IT'S A GOOD EPITOPE FOR OUR AAC. WE HAVE GLYCOSYLATEDDED PROTEINS WE HAVE LCA, PEP TID O GLYCAN AND WILD-TYPE ACID SO GIVEN THE EFFECTS PLOT SHIFT, WE CHOSE TO USE THE WILD TECH EPITOPE AND SUGSO JUST A LITTLE PRIMER ON WHAT [INDISCERNIBLE]-ACID, IT'S BOUND TO THE PEP TID O GLYCAN OF THE GRAM CELL WALL HERE, SHOWN HERE OF MRSA. AND SO THE BINDING IS MEDIATED BY TAG O GLCNAC, AND THE GLUCOSAMINE IS APPENDED TO IN 2 ORIENTATIONS. BY 2 SEPARATE GLUE MARIOUS COSILLEGALS TRANSFERASES WHICH APPEND THE GLUCOSAMINE IN THE BETA BOND AND TAR M IN THE ALPHA BOND AND WE HAVE ANTIBODY AGAINST BOTH FORMS OF THE GLYCOSYLATEDDED WTA. AND SO, WE WERE ABLE TO SHOW THAT THIS ANTIBODY WAS SPECIFIC USING GENETICS HERE. WE SHOW THAT THE ANTIBETTA WTA BINDS TO WILD-TYPE AND TO BE ALPHA ALSO BINDS TO WILD-TYPE BUT CAN YOU SEE HERE THAT IN THE M-MUTE ANT, WE NO LONGER SEE, THE FACTS SHIFTUTESSING THE ALPHA ANTIBODY AND THEN THE MUTANT WE SEE THE CONVERSION. IF I TOLD YOU THE ABUNDANCE OF THE EPITOPE ON THE SURFACE OF THE ANTIBODY,OT SURFACE OF THE BACTERIA WILL BE IMPORTANT BECAUSE IT DICTATES THE NUMBER OF ANTIBIOTIC THAT WE CAN COAT THE BUG WITH. SO FOR THAT REASON WE ACTUALLY CHOSE THE BETA WTA ANTIBODY BECAUSE WHEN WE MEASURE THE BINDING SITES PER BACTERIUM, WE FOUND THERE ARE 50,000 BINDING SITES FOR THE ABET BODY AND ONLY 16 THIS HAPPENED SITES FOR THE ALPHA: AND ALSO BETA, WTA OR TAR S THAT APPENDS BETA WTA IS HIGHLY CONSERVED ACROSS ALL STAFF AUREUS. SO WITH 50,000 BINDING SITES O THEORETICALLY 1 CAN COAT PER BACTERIUM A HUNDRED THOUSAND COPIES OF RIFA LOG. SO WHAT ABOUT THE LINKER? WELL, WE WANT TO SHOW THAT THIS IS CLEAVABLE IN VIVO AND STABLE INVITRO. SO WHAT WE HAVE DONE HERE IS USE THE FRAC TECHNIQUE WHERE THE ALEXIA 488 IS QUENCHED BY TAMARA, AND IT'S ONLY ABOUT CLEAVAGE OF THIS VALVE LINKER THAT WE WILL SEE THE GREEN FLUORESCENCE. SO WE THEN HAVE MACROPHAGE THAT IS BEING--HAVE INTERNALIZED THE STAFF AUREUS, AND YOU CAN SEE THE ABILITY OF 5 MINUTES OF INFECTION, WE BEGENERATED TO SEE THE GREEN FLUORESCENCE INSIDE THE CELL SHOWING THAT THIS LINKER IS INDEED CLEAVED INTRACELLULARLY. AND IT IS REALLY THE CLEAVAGE OF THE LINKER THAT RELEASES ACTIVE ANTIBIOTIC BECAUSE YOU CAN SHOW HERE THAT THIS AAC HAS NO ACTIVITY AGAINST STAPH AUREUS, IT ONLY HAS ACTIVITY WHEN THE CATAPP SIN IS ENTERED IN THE AREA. SO YOU CAN SEE THAT CLEAVABLE AAC IS ABLE TO CLEAR INFECTION IN MACROPHAGE BUT WHEN WE ALTER THE ALANINE TO A D ALANINE WE NO LONGER--THE LINKER IS NO LONGER CLEAVABLE AND THE YOU SEE THE NONCLEAVABLE AAC DOES NOT KILL BACTERIA WITHIN MURINE MACROPHAGE: THIS KILL SUGGEST NOT SPECIFIC TO THE MOUSE BECAUSE WE HAVE ALSO TESTED MULTIPLE HUMAN CELL TYPE AND THE DATA IS CONSISTENT WITH THE ACTIVITY OF THIS AAC IN MACROPHAGES AS WELL AS IN EPITHELIAL AND ENDOTHELIAL CELLS. SO THIS IS ALL IN GOOD. WE HAVE ACTIVITY INVITRO. THE QUESTION IS THAT CAN WE SHOW ACTIVITY IN VIVO? NOW IN ORDER FOR THIS AAC TO WORK, WE WANT TO SHOW THAT IT IS ACTUALLY BETTER THAN THE ANTIBIOTIC STANDARD OF CARE THAT'S BEING USED BECAUSE THAT'S THE UNMET NEED THAT WE WANT TO TACKLE. AND SO, FIRST WE WANT TO SAY WHAT IS THE ANIMAL MODEL THAT CAN HELP US DEMONSTRATE THIS SUPERIORITY AND SO, I KIND OF ALLUDED TO THIS DAT WHEN I SHOWED YOU THAT THE TREATMENT ANIMAL MODEL USING VANCO MICEIN AND MICE TREATED TO VANCOMICEIN TREATED LIKE 1 HOUR POST INFECTION, IT'S VERY EFFICACIOUS. BUT IF YOU NOW BEGIN TO DELAY TREATMENT AT WHICH POINT MANY OF THE BACTERIA ALREADY ENTERED INTO CELLS, YOU CAN SEE HERE THAT BY 24 HOURS POST INFECTION VANCO CAN ONLY HAVE A MINIMAL IMPACTED OF A LOT KILLING. IT'S NO LONGER ABLE TO CLEAR INFECTION. SO WE ASKED THEN WOULD THE AAC ABLE TO CLEAR INFECTION WHEN YOU BEGIN TREATMENT AT 24 HOURS POST INFECTION WHEN VANCO DOES NOT WORK AS WE THINK WOULD MIMIC THE CLINICAL CONDITION. SO HERE'S THE EXPERIMENT. SO WE HAVE MICE THAT WERE TREATED WITH SALINE, MICE THAT WERE TREATED WITH THE ANTIBODY ALONE. THE ANTIBODY USED THROUGH AAC, THAT IS MINIMAL EFFECT, VACIN HCO IS PARTIALLY EFFECTIVE BUT THE AAC IS NOW ABLE TO CLEAR INFECTION. LET SO DEMONSTRATING THAT IT IS SUPERIOR TO VACIN, COMICEIN IN THIS ANIMAL MODEL. WE ALSO TEST THAD THE ABILITY OF THIS AAC SUPERIORITY IN THE MODEL, LIKE INDEPT O MICEIN. SOPHISTICATED IF WE BEGIN TREATMENT POST INFECTION, WE SEE THIS VARIABLE EFFECT, RIGHT? THERE IS SOME TREATMENT EFFECT BUT OTHERS ARE NOT WELL CONTROLLED FOR THE DAPTOMYCIN, IT IS CLEARED AND WE SEE THERE IS NO NEGATIVE-NEGATIVE DRUG INTERACTION. THERE THE DATA ARE ENCOURAGING BUT 1 CAVEAT IS THAT MICE ARE NOT NATURAL COLONIZERS--ARE NOT NATURALLY COLONIZED WITH STAPH AUREUS, SO THEY DON'T HAVE NTWA ANTIBODY IN THE SERUM. MANY OF US AREICALONNIZED WITH THE STAPH AUREUS AND MANY DO HARBOR NTWA IN OUR SYSTEM. SO THAT COULD COMPETE AND DETERMINE THE AMOUNT OF ANTIBODY THAT COULD BE FOUND IN HUMAN SO WE MIMIC THIS ENVIRONMENT NOW BY FIRST TREATING THE MICE, NOW WE USE SKID MICE WITH IGIV AT 10 MICRO GRAM PER MILL AND SO NOW THIS MICE HAVE NTWA ANTIBODY IN THEM. AND THEN WE REPEAT THE EXPERIMENT AS I'VE SHOWN YOU BEFORE AND YOU CAN SEE HERE THAT EVEN IN THE PRESENCE OF COMPETING WTA ANTIBODY, THIS [INDISCERNIBLE] ALSO WORKS WELL. AND WE ALSO ADDED THE CONTROL, IN THIS CASE ANTIMRSAAAC, AND YOU SEE AND YOU WONDE-PRESCRIBING WHY? WELL STAPH AUREUS, AS YOU KNOW CONTAINS PROTEIN A AND IT WILL BIND ANY ANTIBODY BUT OBVIOUS LEAP, THE AMOUNT OF PROTEIN A ON THE STAPH AER EUS IS MAGNITUDES LESS AND WE SEE THAT THE EFFICACY WE SEE HERE IS DUE TO ANTICMTAA BINDING TO PROTEIN A. BUT I DID TELL YOU THAT THE ANTIBIOTIC THAT WE IDENTIFIED, IT'S A VERY POTENT AND EXCITING MOLECULE IN AND OF ITSELF. MAYBE THE ANTIBODY ITSELF IS GOOD ENOUGH FOR ACTHAT WE SEE. SO THIS IS--ACTIVITY THAT WE SEE. SO THIS IS EXPERIMENT TO TEST THAT IDEA IS SO HERE SAME EXPERIMENT AS I SHOWED PREVIOUSLY EXCEPT THAT WE HAVE TRIED TO ADDRESS 2 OTHER QUESTIONS: IF WE USE NONCLEAVABLE AAC, WE ARE NOT ABLE TO SEE EFFICACY. IF WE JUST USE THE ANTIBIOTIC ALONE EITHER A SINGLE TREATMENT OR DAILY TREATMENT WE ALSO DO NOT SEE EFFICACY. SO THIS BRINGS HOME AN IMPORTANT LESSON. THAT WE THINK THAT THIS PLATFORM COULD PROVIDE FOR ANTIBIOTIC. FOR ANTIBIOTIC THAT CANNOT ENTER CELLS VERY WELL OR ANTIBI THETIC THAT HAS POOR FARM CO KINEETICS. BY LINKING ANTIBIOTIC TO THE ANTIBODY WE CAN NOW HAVE THE FARM COKINETTIC AS THE ANTIBODY OF THE DRIVER. SO WHAT I TOLD YOU HERE TODAY IS THAT WE HAVE A NOVARTIS AND HE WILL POTENT EFFECT EVALUATION PROCESS TREATMENT AGAINST MRSA. THE EFFICACY OF AAC SUGGESTS THAT INTRACELLULAR INFECTION DOES CONTRIBUTE TO FAILURE OF SOC ABET BIOTICS. I SHOWED YOU THAT VANCOMICEIN AND DAPTOMICEIN AND ANTIBODY OF THE AAC PROVIDES SUPERIOR PK AND THIS AAC IS PRESENT IN THE HUMAN SYSTEM AS A PRO DRUG THAT IS CONSTANTLY CIRCULATING IN THE ASPECT BODY AND WHENEVER THERE IS STAPH AUREUS, IT WILL BIND TO STAFF OR DRIVE IT INTO THE CELL AND ONLY INSIDE THE CELL IS ANTIBEING RELEASED. AND SO WE ARE HOPEFUL THAT THIS PLATFORM WILL ENABLE THE USE OF MORE TOXIC ANTIBIOTIC BECAUSE OF THE DILUTION EFFECT THAT I TOLD YOU ABOUT AS AS IT WILL BE ABLE TO SPARE HUMAN MICRO BIOME AND WE THINK THAT THIS IS A PROOF OF CONCEPT STUDY THAT SUGGESTS THAT THE AAC PLATFORM COULD BE USED TO TREAT OTHER BACTERIAL INFECTIONS AS WELL. SO 1 THING THAT I LIKE TO KIND OF MAKE A PLEA TO SCIENTIFIC COMMUNITY IS THAT OFTEN TIMES WE GET RID OF ANTIBIOTIC THAT ARE EITHER TOXIC OR HAVE NO GOOD PK BECAUSE IT'S NOT POSSIBLE TO DEVELOP THAT MOLECULE AT THE STAND ALONE ANTIBIOTIC TO TREAT INFECTION. MENT BUT THOSE MOLECULES COULD POTENTIALLY RESCUED BY USING THIS PLATFORM. SO IF YOU ARE AWARE OF ANY ANTIBIOTIC THAT IS POTENT BUT HAS POOR PK AND MAYBE EVEN A LITTLE TOXIC, WE MIGHT STILL BE ABLE TO TEST WHETHER THAT ANTIBIOTIC COULD BE USED IN THIS CONTEXT. AND WOULD BE AN ANTIBIOTIC AGAINST GRAM NEGATIVE BACTERIA OR AGAIN TB. SOPHISTICATEDY WITH THAT I WANT TO THANK AS YOU CAN APPRECIATE THE LARGE AMOUNT OF WORK THAT'S NEEDED TO BUILD AND FIND THIS ANTIBODY SO THE INFLUENZA TEAM SHOWN IN GREEN ARE THE INDIVIDUALS WHO ARE SPEAR HEADING THE PROGRAM. WE WANT TO THANK THE CDC FOR TESTING SOME OF THE [INDISCERNIBLE] AND FOR THE VOLUNTEERS AND PATIENTS. THIS MOLECULE IS IN THE PHASE 2 B CLINICAL TRIAL. SO WE'RE AWAITING ANXIOUSLY TO HAVE THE DATA UNBLINDED IN THE FUTURE AND FOR THE AAC PROGRAM [INDISCERNIBLE] TAKE OTHER LEADERS ON TED BIOLOGY, CHEMISTRY AND CONJUGATION OF THE EXPERIMENT, AND SOVEY DID MOST OF THE EXPERIMENT THAT I PRESENTED AND I WANT TO ACKNOWLEDGE RICK WHO HAD THE FORESIGHT TO START INFECT YOWS DISEASE RESEARCH IN GENENTECH, UTILIZING THE AND LEVERAGING THE KNOW-HOW ON ANTIBODY AND USE IT FOR INFECTION DISEASE. WE DO HAVE OTHER PROGRAMS IN SMALL MOLECULE AND HOPEFULLY IN THE NEAR FUTURE WE WILL BE ABLE TO TELL YOU ABOUT THEM AS WELL AND WITH THAT I TANK YOU FOR YOUR ATTENTION AND I--I THANK YOU FOR YOUR ATTENTION AND I AM HAPPY TO TAKE QUESTIONS. [ APPLAUSE ] >> [INDISCERNIBLE] FOR THE STAPH AUREUS, WE HAVE MANY LINKAGE SO WE HAVE MANY [INDISCERNIBLE] I WAS WONDERING IF IN THE DIFFICULT COMPLEX [INDISCERNIBLE] PLASMA, HOW DOES THE SURVIVAL OF THOSE SECTION OF THE AACs THAT COULD BE AVAILABLE FOR [INDISCERNIBLE] LIGATION IS KILLING THE BACTERIA. >> YEAH, SO WE HAVE SELECTED THE VALVES THAT LINGER SO WE EXPERIENCE AND THIS IS WHERE WE ACTUALLY DREW OUR EXPERIENCE ON THE AAC FROM THE ANTIBODY DRUG CONJUGATE PROGRAM IN ONCOLOGY. SO VAL-SET HAS BEEN TESTED IN PATIENTS AND ARE STABLE IN HUMAN SERUM. SOPHISTICATEDY WE KNOW THAT AND THEN WE HAVE ALSO TESTED THE STABILITY OF THIS LINKER TO PROTEASES THAT HAVE BEEN PRODUCED BY BACTERIA AND WE SHOWED THAT THIS LINKER IS STABLE. >> SO THE REQUESTY IS WHAT PRESENTATION BREAKS DOWN-- >> WE'RE GOING TO HAVE VERY FEW QUESTIONS-- >> SORRY. >> MY OVERALL QUESTION IS THE RIFALYNN IF I AM CORRECT, THE PRONOUNCEIATION, YOU MENTION SOMETHING INTERESTING REGARDING THE ACIDIC CONDITION THAT YOU INDUCE BUT IT WAS A LITTLE BIT ISOLATED FROM THE EXPERIMENT TO EXPERIMENT. THE QUESTION IS WHETHER THE ACIDIFICATION OF THE ENVIRONMENT CAN HELP INDUCE HIDROLLISS AND ALSO HIDE ROLIZES OF THE EO GOLGI AND IN THE PLASMID--RIBOSOMAL ENZYMES FOR RECYCLING AND CHEWING DOWN PROBABLY THE BACTERIA? >> LET ME SEE IF I UNDERSTAND YOUR QUESTION CORRECTLY. SO WHETHER THE PROTEASE WAS PRODUCED BY THE HOST ITSELF IS SUFFICIENT TO-- >> ACIDDIVEIFICATION. >> ACIDIFICATION. >> SO ACIDIFICATION ITSELF IS NOT THE PROCESS FOR THE LINKER, THE LINKER THAT IS TO BE CLEAVED BY C ATAPSIN, WHICH IS PART OF THE ENDOSEDDIC PROCESS, SO THE ACIDIFICATION HERE IS THE ENVIRONMENT IN WHICH THE CATAFSIN IS RELEASED SO WE NEED THE ANTIBIOTIC TO BE ACTIVE UNDER THAT ENVIRONMENT. DID I ADDRESS THAT QUESTION? >> THAT'S FINE. THAT'S FINE. WE CAN TALK. >> SO BACK TO THE--SORRY, I WAS WONDERING WHAT'S THE RELATIVE ENFORCEMENT OF [INDISCERNIBLE] VERSUS DIRECT ACTIVITY OF THE ANTIBODY? THE REASON I ASK IS IF YOU HAVE A RESISTANCE VIRUS WHERE THE ANTIBODY CAN STILL BIND, IS THAT VIRUS STILL PROTECTED AGAINST IN VIVO? >> YES, SO I'VE NOT SHOWN THE DAT AWE HAVE THE DATA SPECIFICALLY FOR THE INFLUENZA B: SO WE HAVE ISOLATED RESISTANT STRAIN THAT ABOLISHS BINDING AT ACIDIC Ph BUT RETAINS BINDING AT NEUTRAL Ph. WE THEN INFECTED MICE WITH THIS RESISTANT MUTE ANT AND SHOWED THAT THE--MUTANT AND SHOWED THAT THE ASPECT BODY CAN PROTECT THE MOUSE COMPLETELY. SO THAT SUGGESTS TO US THAT UNDER THAT CONDITION THE ACC IS PLAYING THE MAJORITY OF THE ROLE IN CLEARING INFECTION. >> THANK YOU SO MUCH WONDERFUL TALK. I WAS WONDERING, SPECIFIC QUESTION, HOW WELL THESE ANTIBODIES PERFORM IN LOCALIZED INFECTION MODELS? YOU KNOW WHERE YOU MAY HAVE THINGS LIKE BIOFILMS AND ISSUES LIKE THAT? AND THEN WHAT CAN WE DO TO MAKE MONOCLONAL ANTIBODIES AND,A ACs POTENT AGAINST THE LOCALIZED TARGET. >> YES, SO WE HAVE NOT DIRECTLY ADDRESSED THAT QUESTION AND MY REQUESTS IS THAT FOR THE BIOSTUDIES OF MULTIPLE ENDOCRINE IS NOT GOING TO BE VERY EFFECTIVE. BUT TO THE POINT ABOUT HOW DO WE ELIMINATE BIOFIRM, SO WE THINK ABOUT THIS AS A PLATFORM OR IF YOU WANT TO GO SPECIFICALLY TO BIOFILM 1 COULD THINK ABOUT THE POSSIBILITY OF FINDING ANTIBODY THAT BINDS TO BIOFILM, RIGHT? THE SUBSTRATE OF THE BIOFILM. ONE CAN CHANGE THE LINKER TO A LINKER THAT'S CLEAVABLE BY AN ENZYME THAT'S PRODUCED BY THE BACTERIA. AND THE BIOFILM. SO NOW EFFECTIVELY WHAT YOU HAVE DONE IS CONCENTRATE THE ANTIBIOTIC TO THE BIOFILM AND RELEASED AN ACTIVE ANTIBIOTIC AT THE BIOFILM SITE. SO I THINK THE CHALLENGE HERE WITH AAC IS THAT THE SITE OF INFECTION WILL BE IMPORTANT BECAUSE ANTIBODY IS BEING DISTRIBUTED BY THE BLOOD STREAM AND SO, IN IN AREAS WHERE ANTIBODY DOES NOT GET TO WOULD BE A CHALLENGE TO THE AAC CONCEPT. BUT 1 CAN THINK ABOUT HOW WE MODIFY THE ANTIBODY AS AS LINKER FOR BIOFILM AND OTHER DISEASES. >> THANK YOU. >> HAVE YOU EVER OBSERVED A DRUG RESISTANCE AGAINST AAC COMPARED TO WITH [INDISCERNIBLE]--THANK YOU. >> SO VERY GOOD QUESTION. WE HAVE. IT'S VERY DIFFICULT TO DO THE EXPERIMENT WHERE WE SCREEN FOR RESISTANCE AGAINST THE AAC AS A WHOLE. WE HAVE DONE--ISOLATE, JUST DO THE TYPICAL RESISTANCE STUDY WITH THE DRUG ALONE AND WE CAN FIND RESISTANCE OBVIOUSLY. FELT AT LEAST IN THE ANIMALS IN WHICH WE HAVE BEEN INFECTED THIS STAPH AUREUS, WE HAVEN'T RESISTED IT DOESN'T MEAN WE HAVEN'T RESIST, IT MEANS WE HAVEN'T FOUND THEM. THE DIFFICULTLE IN FINDING RESISTANCE IS BECAUSE UNLIKE THE ANTIBIOTIC THAT'S FREELYY FLOATING IN THE SYSTEM SO IT'S BEEN EXPOSED TO 10 TO THE 16 TO THE 7 BACTERIUM, AND SO YOU HAVE NOW, THE RESISTANCE IN 1 TO THE 10 RESISTANT BACTERIA AND YOU'RE ONLY DRIVING HUNDREDINGS OF BACTERIA INSIDE 1 CELL SO THE PROBABILITY OF THE IT BEING RESISTANT IS GOING TO BE MUCH SMALLER BUT WE ARE CONCERNED ABOUT THAT AND IN ORDER FOR THIS BE EFFICACIOUS AND FOR THE--FOR WHAT WE DESIGNED IT THE AAC TO BE DO IN OUR CLINICAL TRIAL WE ARE GOING TO BE TREATING PATIENTS WHO ARE ALREADY TREATED WITH THE STANDARD OF CARE AND NOT PROGRESSING WELL IN THE PRESENCE OF THE STANDARD OF CARE. AND THEN WE ADD THE AAC TO HELP THIS 20% OF PATIENTS ARE GOING TO POTENTIALLY DIE OF THE INFECTION AND SO EVEN IF THERE IS RESISTANCE THERE ARE GOING TO BE 2 MECHANISMS OF RESIEVANCE THAT THE BUG WILL DEAL WITH. SO WE DID NOW TEST THE RESISTANT OR THE RESISTANT MUTE ANT. WE ARE NOT--THEY DO NOT SO INCREASED RESISTANCE TO ADAPTER OR VANCO OR ANY OTHER STANDARD OF CARE OR ANTIBIOTIC. SO WITHIN THE SITUATION WHERE THIS AAC WILL WORK, WILL BE USED. WE DO NOT THINK THAT THE RESISTANCE WILL BE AS CRITICAL. >> OKAY, THANK YOU VERY MUCH. THERE IS A REACCEPTION THAT FOLLOWS THIS SEMINAR RIGHT ACROSS THE HALL IN THE LIBRARY AND I THANK YOU ALL FOR COMING AND MAN-WAH FOR SUCH A BEAUTIFUL PRESENTATION. >> THANK >> [ APPLAUSE ] YOU. >> [ APPLAUSE ]