Reinventing the Gene Vaccine com Andrew Geall

Full Transcript

https://www.youtube.com/watch?v=cesLJwOePMk

[00:41] uh I'm going to switch to English
[00:43] uh I'm going to switch to English because we have an an international speaker today uh Angel
[00:46] because we have an an international speaker today uh Angel
[00:49] speaker today uh Angel Gil uh has been very generous and given us lots of lectures this is his fifth lecture in three days so uh
[00:52] Gil uh has been very generous and given us lots of lectures this is his fifth lecture in three days so uh
[00:56] us lots of lectures this is his fifth lecture in three days so uh
[01:00] Lecture in three days so uh he thought he would just have fun uh.
[01:03] He thought he would just have fun uh coming to Brazil I I I hope he doesn't.
[01:06] Coming to Brazil I I I hope he doesn't get scared uh but Andrew Gil is um uh.
[01:12] Scared uh but Andrew Gil is um uh co-founder and chief development officer.
[01:14] Co-founder and chief development officer of replicate biosciences he has over 20 years of experience in developing drugs.
[01:18] Years of experience in developing drugs delivery system uh as a pioneer in mRNA vaccines and nucleic acid delivery uh he.
[01:24] Vaccines and nucleic acid delivery uh he currently serves as Chief development officer at replicate uh which he co-founded.
[01:31] Currently serves as Chief development officer at replicate uh which he co-founded.
[01:34] Prior to replicate uh he was Chief scientific officer at Precision Nano systems.
[01:37] Chief scientific officer at Precision Nano systems he also served as vice president of formulations analytics and chemistry at avidity bioscience and RNA uh vaccine platform leader at Novartis uh.
[01:40] Nano systems he also served as vice president of formulations analytics and chemistry at avidity bioscience and RNA uh vaccine platform leader at Novartis uh.
[01:42] He also served as vice president of formulations analytics and chemistry at avidity bioscience and RNA uh vaccine platform leader at Novartis uh.
[01:45] President of formulations analytics and chemistry at avidity bioscience and RNA uh vaccine platform leader at Novartis uh.
[01:49] Chemistry at avidity bioscience and RNA uh vaccine platform leader at Novartis uh.
[01:52] RNA uh vaccine platform leader at Novartis uh he was at Novartis for over seven years uh.
[01:55] Novartis uh he was at Novartis for over seven years uh.
[01:58] He was at Novartis for over seven years uh he holds a pharmacy degree from the University of ba in.
[02:02] degree from the University of ba in United Kingdom and completed.
[02:04] United Kingdom and completed pre-registration training in a septic.
[02:06] pre-registration training in a septic dispensing unit at St Helen's uh nossle.
[02:10] dispensing unit at St Helen's uh nossle trust hospital he has a PHD uh at the.
[02:15] trust hospital he has a PHD uh at the University of bath to in nonviral uh.
[02:18] University of bath to in nonviral uh Gene delivery.
[02:21] Gene delivery he also he's also a a rower and he almost went to the Olympics.
[02:25] rower and he almost went to the Olympics and if that happened um maybe the his.
[02:29] and if that happened um maybe the his history of RNA vaccines would be quite different.
[02:32] history of RNA vaccines would be quite different because he he played a pivotal.
[02:37] different because he he played a pivotal role in developing all these Delivery Systems that we use today that modna.
[02:41] role in developing all these Delivery Systems that we use today that modna used that F used and um.
[02:44] used that F used and um uh now there is a Judicial litigation on.
[02:50] uh now there is a Judicial litigation on this patents that because companies made.
[02:55] this patents that because companies made billions during the pandemic and he is
[03:02] billions during the pandemic and he is in the center of five patents that is in the center of five patents that is being litigated so uh definitely the being litigated so uh definitely the story of the the covid would be different um if his uh rowing in career launched uh for good uh and he's also he still Roar every day uh and he's a coach of a rowing uh girls girls rowing team and also he's a scuba div passionate so you will see some photos at least the last photo do yeah the last one is uh a turtle that he uh took himself so angel thank you so much for all this day we have been a lot of fun with you here uh learn a lot so thank you very much thank you Elton and you know I've this is day three presentation number five I had a great time it's been awesome um I I love coming in teaching
[04:04] awesome um I I love coming in teaching the students trying to inspire them to.
[04:07] the students trying to inspire them to hand the bat over uh it's your turn uh.
[04:10] hand the bat over uh it's your turn uh so this is to me I have this is my.
[04:13] so this is to me I have this is my second favorite type of presentation the.
[04:16] second favorite type of presentation the first one is my career Journey with the.
[04:18] first one is my career Journey with the students and we did that one yesterday.
[04:19] students and we did that one yesterday and that was a lot of fun but this is my.
[04:22] and that was a lot of fun but this is my this is my 16-year story with um DNA and.
[04:26] this is my 16-year story with um DNA and RNA but trying to reinvent the gene.
[04:28] RNA but trying to reinvent the gene vaccine and and overcome some Monumental.
[04:32] vaccine and and overcome some Monumental hurdles and this is the story of you.
[04:34] hurdles and this is the story of you know how we achieved it um I've worked.
[04:38] know how we achieved it um I've worked with probably close to a thousand.
[04:40] with probably close to a thousand colleagues now on on this project um.
[04:43] colleagues now on on this project um certainly hundreds and you know I'm one.
[04:46] certainly hundreds and you know I'm one individual I get to tell the story but.
[04:48] individual I get to tell the story but you know as I give the seminar I have.
[04:50] you know as I give the seminar I have memories of all of these people I work.
[04:52] memories of all of these people I work with they all work super hard um it's.
[04:55] it's not just me it's a lot of work from a.
[04:57] not just me it's a lot of work from a lot of people so please appreciate.
[04:59] apprciate the work that they put into um.
[05:02] apprciate the work that they put into um so this is the journey and you know.
[05:04] so this is the journey and you know through the slides and through data um.
[05:07] through the slides and through data um you you'll you'll see the journey there.
[05:08] you you'll you'll see the journey there a few pictures of faces too um so it begins in 2008 this is when I took over.
[05:12] a few pictures of faces too um so it begins in 2008 this is when I took over uh well started the RNA vaccine program at nartis and we'll spend a few slides.
[05:16] begins in 2008 this is when I took over uh well started the RNA vaccine program at nartis and we'll spend a few slides.
[05:18] uh well started the RNA vaccine program at nartis and we'll spend a few slides getting on how how did that happen and.
[05:20] at nartis and we'll spend a few slides getting on how how did that happen and why did it happen um and then the.
[05:22] getting on how how did that happen and why did it happen um and then the Journey Begins I measure performance of a vaccine that's everything does it is.
[05:25] why did it happen um and then the Journey Begins I measure performance of a vaccine that's everything does it is.
[05:27] Journey Begins I measure performance of a vaccine that's everything does it is does it have efficacy neutralizing antibodies is it tolerated what's the safety profile how many doses do you need but the main question for me is is it competitive does it work the same or better than a conventional technology.
[05:30] a vaccine that's everything does it is does it have efficacy neutralizing antibodies is it tolerated what's the safety profile how many doses do you need but the main question for me is is it competitive does it work the same or better than a conventional technology.
[05:32] does it have efficacy neutralizing antibodies is it tolerated what's the safety profile how many doses do you need but the main question for me is is it competitive does it work the same or better than a conventional technology.
[05:34] antibodies is it tolerated what's the safety profile how many doses do you need but the main question for me is is it competitive does it work the same or better than a conventional technology.
[05:37] safety profile how many doses do you need but the main question for me is is it competitive does it work the same or better than a conventional technology.
[05:40] need but the main question for me is is it competitive does it work the same or better than a conventional technology.
[05:43] it competitive does it work the same or better than a conventional technology because if it doesn't um we're not going to displace anything and so the.
[05:45] better than a conventional technology because if it doesn't um we're not going to displace anything and so the Journey's always been about getting up this Innovation curve uh and being able to perform or outperform conventional technology.
[05:48] because if it doesn't um we're not going to displace anything and so the Journey's always been about getting up this Innovation curve uh and being able to perform or outperform conventional technology.
[05:50] to displace anything and so the Journey's always been about getting up this Innovation curve uh and being able to perform or outperform conventional technology.
[05:53] Journey's always been about getting up this Innovation curve uh and being able to perform or outperform conventional technology.
[05:56] this Innovation curve uh and being able to perform or outperform conventional technology.
[05:58] to perform or outperform conventional technology.
[06:00] technology going fast is great in a pandemic but that's all it counts for in.
[06:03] going fast is great in a pandemic but that's all it counts for in.
[06:06] pandemic but that's all it counts for in the end.
[06:09] uh in the real world when we're in non-pandemic situations so you got to compete with conventional technology.
[06:11] so you got to compete with conventional technology.
[06:14] so you know I'll take you through the journey at nartis.
[06:15] we got through the efficacy threshold in the animal models.
[06:17] I'll show you some of that data.
[06:20] some of the work is very interesting and pioneering.
[06:22] um you know we weren't the first to go into clinical trials with RNA.
[06:25] that was madna they went in in 2015 with linear base modified RNA in an LMP in an influenza trial in a pandemic setting.
[06:27] a single antigen data was okay.
[06:29] uh it wasn't going to compete with conventional technology.
[06:34] uh but it was the first real clinical trial showing success.
[06:37] then we had covid not going to spend much time here.
[06:41] the early successes with linear RNA but then there was a self-amplifying RNA from uh Arcturus that took a lot longer to come through.
[06:44] but it's showing better durability.
[07:08] but it's showing better durability uh than the linear systems and then I'm
[07:10] than the linear systems and then I'm going to take you through some pivotal
[07:12] going to take you through some pivotal data that we just released from uh my
[07:14] data that we just released from uh my company uh replicate bioscience in San
[07:16] company uh replicate bioscience in San Diego my passion is much longer RNA uh
[07:20] Diego my passion is much longer RNA uh self-replicating
[07:21] self-replicating systems so my journey really began in
[07:26] systems so my journey really began in 2002 was my first job off my posto I
[07:29] 2002 was my first job off my posto I educated in England and I came over on a
[07:31] educated in England and I came over on a postto I actually worked on malaria uh
[07:34] postto I actually worked on malaria uh learned to be a parasitologist looking
[07:36] learned to be a parasitologist looking at transport Pathways of small molecules
[07:38] at transport Pathways of small molecules in arthrit infected with malaria um and
[07:43] in arthrit infected with malaria um and I managed to uh get a job uh at a
[07:47] I managed to uh get a job uh at a company in San Diego called vial the DNA
[07:50] company in San Diego called vial the DNA vaccine company um Phil falgner was one
[07:53] vaccine company um Phil falgner was one of the founders we delivered DNA with
[07:55] of the founders we delivered DNA with lipoplex Delivery Systems and I took in
[07:58] lipoplex Delivery Systems and I took in a period of four years is four clinical
[08:01] a period of four years is four clinical strategies forward through phase one and
[08:03] strategies forward through phase one and phase two scaling up the material in the
[08:06] phase two scaling up the material in the clean rooms also in the on the bench as
[08:09] Clean rooms also in the on the bench as a research scientist so doing prototypes.
[08:11] A research scientist so doing prototypes and then getting it in the clean room.
[08:13] And then getting it in the clean room making phase one material releasing it.
[08:15] Making phase one material releasing it scaling it further and those delivery.
[08:18] Scaling it further and those delivery systems were very very complex very.
[08:20] Systems were very very complex very difficult to scale 2000 uh and six I.
[08:24] Difficult to scale 2000 uh and six I decided to move on move the family uh to.
[08:28] Decided to move on move the family uh to Boston and I started working um in.
[08:33] Boston and I started working um in uh pharmaceutical profiling group and my.
[08:36] Uh pharmaceutical profiling group and my job was to design the delivery.
[08:39] Job was to design the delivery strategies for short interfering RNA in.
[08:43] Strategies for short interfering RNA in 2006 fire and mellow received the Nobel.
[08:47] 2006 fire and mellow received the Nobel uh prize for medicine for the discovery.
[08:50] Uh prize for medicine for the discovery of RNA interference and so they were.
[08:52] Of RNA interference and so they were able to show that these duplexes of.
[08:54] Able to show that these duplexes of short RNA uh if they're coded correctly.
[08:57] Short RNA uh if they're coded correctly you can get them into cell they'll.
[08:59] You can get them into cell they'll buying in a risk complex and this will.
[09:02] Buying in a risk complex and this will uh.
[09:03] Uh ablate a specific sequence of messenger.
[09:06] Ablate a specific sequence of messenger RNA so it's a NE knockdown technology uh.
[09:09] RNA so it's a NE knockdown technology uh we were at that time focused on liver.
[09:11] we were at that time focused on liver delivery and rare muscle rare diseases.
[09:15] delivery and rare muscle rare diseases within the liver the lead delivery.
[09:17] within the liver the lead delivery system was this lipid.
[09:19] system was this lipid nanoparticle and RNA was going to change the world.
[09:22] nanoparticle and RNA was going to change the world I was on board I was started my job and it really wasn't ready it.
[09:27] my job and it really wasn't ready it wasn't until 2008.
[09:29] wasn't until 2008 that this LMP was finally ready to transition.
[09:32] that this LMP was finally ready to transition it was a product treating people with ram liver diseases.
[09:35] transition it was a product treating people with ram liver diseases what was wrong.
[09:38] people with ram liver diseases what was wrong uh the chemistry on the SI RNA wasn't there.
[09:41] uh the chemistry on the SI RNA wasn't there there was a lot of off-target knockdown effects.
[09:42] wasn't there there was a lot of off-target knockdown effects the potency wasn't there the stability wasn't there.
[09:45] the potency wasn't there the stability wasn't there.
[09:47] wasn't there the stability wasn't there the lipid nanoparticles were highly toxic in the animal models.
[09:49] the lipid nanoparticles were highly toxic in the animal models and we were scaling.
[09:52] toxic in the animal models and we were scaling we were screening rapidly to find more and more potent LMP delivery system.
[09:56] scaling we were screening rapidly to find more and more potent LMP delivery system to get the dose down lower so that we could thread the needle a therapeutic index.
[09:58] system to get the dose down lower so that we could thread the needle a therapeutic index get below the toxicity.
[10:01] that we could thread the needle a therapeutic index get below the toxicity and see efficacy.
[10:03] therapeutic index get below the toxicity and see efficacy.
[10:05] get below the toxicity and see efficacy I like the lipid nanoparticles.
[10:09] efficacy I like the lipid nanoparticles.
[10:12] efficacy I like the lipid nanoparticles because for the first time they were a scalable liposome technology for the.
[10:17] scalable liposome technology for the first time we had an ATS scale way of mixing uh um these particles processing them at scale and so I was focused internally at notis on building novel uh lipids filing patents scaling the process and in.
[10:35] process and in 2008 um somebody approached me from the vaccine division this is Christian mandle uh Chiron was acquired by nartis vaccines in 2006 and in 2008 they relocated um the division to Cambridge in Boston.
[10:52] relocated um the division to Cambridge in Boston he is an academic uh from the University of Vienna uh and he published this paper uh in pnas where they taken uh a Replicon so a self-amplifying RNA from uh tickborne and seyis TBE um encoded an antigen in it they coated it.
[11:14] encoded an antigen in it they coated it onto gold particles and using Gan gun.
[11:18] onto gold particles and using Gan gun they' gone into a mouse they vaccinated.
[11:20] they' gone into a mouse they vaccinated this mouse 10 or 12 times uh and shown.
[11:24] they could seroc convert the mouse to the antigen using this novel technology.
[11:29] really just a proof of concept it wasn't something that they could develop beyond that first experiment and Gan gun is never used by anyone uh you know since.
[11:39] those times really but Christian was passionate about RNA and he knew delivery was the problem and he wanted a delivery person to run the program and he heard that I'd worked at vial was a delivery person had a history and the DNA vaccines and he wanted to try and encourage me to come and work on the program.
[11:59] the first thing I said to him was you know I've read your papers this interesting but there's no way this is going to work and he said no I we need a you know a scalable delivery technology uh and I talked to him about lipid.
[12:14] uh and I talked to him about lipid nanoparticles uh I told him I needed nanoparticles.
[12:16] uh I told him I needed about a month to go and review the literature on how to make RNA um and so for a month I buried myself in in the historical literature from about the 1980s um back to the 1970s where the biochemists had had to scale enzymatic transcription reactions to make RNA so they could bind them with proteins and do crystal structures from that reading it was obvious to me that we could scale the technology that RNA could be made enzymatically now whether we could do it uh and have stability there were a lot of unknown questions but there was enough there for me to think that this was not that risky you move to switch divisions.
[13:00] now he offered me two posts and a couple of technicians and I was leaving a group of 16 PhD students half of them in Switzerland half of them based in Boston on on this pipe dream my peers thought I was utterly crazy to be
[13:15] Peers thought I was utterly crazy to be moving but I just felt that it was there.
[13:19] Moving but I just felt that it was there was momentum and there was a great opportunity.
[13:21] I love to innovate and I couldn't resist.
[13:24] My wife who said you'll be happy go and so 2008 I left on this dream and vision of reinventing the gene vaccine.
[13:26] Jeff Olmer was also there um he was the the global head of external research and he we wrote together.
[13:30] I think 25 papers describing the technology uh in various peer-review journals and reviews.
[13:33] Jeff was the editor of those I was the writer and then the scientists who contributed dropped in as the middle authors.
[13:37] So these are you know old memories of of of a great time.
[13:40] So 2008 I began and um you know this is now my career history to date and we're going to kind of pinch point on all of these things.
[13:44] We'll the data preclinical data.
[14:17] things we'll the data preclinical data from novatis for seven years uh then.
[14:21] from novatis for seven years uh then Novartis was sold sold off their vaccine.
[14:24] Novartis was sold sold off their vaccine division to GSK and I left I came back.
[14:26] division to GSK and I left I came back to San Diego where I had started and.
[14:28] to San Diego where I had started and where wife is f I did something.
[14:30] where wife is f I did something completely different uh delivering sna.
[14:34] completely different uh delivering sna but this time as a conjugate on an.
[14:35] but this time as a conjugate on an antibody uh I got one slide describing.
[14:38] antibody uh I got one slide describing it there's a whole lecture and seminar.
[14:40] it there's a whole lecture and seminar on that technology too it's been highly.
[14:42] on that technology too it's been highly successful but while I was doing that I.
[14:44] successful but while I was doing that I was Consulting with the RNA vaccine.
[14:47] was Consulting with the RNA vaccine field because of my you unique.
[14:49] field because of my you unique experiences and over be was able to.
[14:52] experiences and over be was able to overcome some challenges uh by working.
[14:55] overcome some challenges uh by working with this company in Vancouver Precision.
[14:57] with this company in Vancouver Precision Nanos systems particularly about.
[14:59] Nanos systems particularly about building out a lipid library and scaling.
[15:02] building out a lipid library and scaling and mixing.
[15:04] and mixing technology I went full-time in the first.
[15:06] technology I went full-time in the first year of covid there raised a large sum.
[15:09] year of covid there raised a large sum of money from the Canadian government to.
[15:11] of money from the Canadian government to build out the platform for a covid.
[15:12] build out the platform for a covid vaccine and build a biomanufacturing.
[15:16] vaccine and build a biomanufacturing center in Vancouver uh for the next.
[15:19] center in Vancouver uh for the next pandemic the company was bought a year.
[15:22] pandemic the company was bought a year in bought by danah herd now they're part.
[15:25] in bought by danah herd now they're part of the organization stiva and I went.
[15:27] of the organization stiva and I went full-time at repli at um which i'.
[15:31] full-time at repli at um which i' co-founded with a friend in.
[15:34] 2020 so at ntis this was um the vision.
[15:40] uh for flu vaccines so this is a.
[15:42] state-of-the-art mamalian cell culture.
[15:45] facility uh it was finally commissioned.
[15:47] in uh.
[15:49] in uh 2012 uh but they were building it.
[15:52] 2012 uh but they were building it operations were beginning it's a massive.
[15:55] facility.
[15:56] um 475.
[15:59] uh square feet uh 900 employees you can.
[16:02] see the fermentation trains here these.
[16:05] are 2,000 liter uh uh fermenters we're.
[16:09] at the top deck when you look down it's.
[16:11] a couple of hundred feet down uh and.
[16:14] then you've got all the associated.
[16:15] Downstream purification uh systems this.
[16:18] is all clean in place uh it's running.
[16:21] is all clean in place uh it's running nonstop and from that facility you know.
[16:24] nonstop and from that facility you know you can produce uh seasonal uh.
[16:27] you can produce uh seasonal uh pre-pandemic and pandemic uh vaccines it.
[16:30] pre-pandemic and pandemic uh vaccines it was a co-development project with the US.
[16:32] was a co-development project with the US government so if there's a pandemic in.
[16:35] government so if there's a pandemic in flu it's designed to produce 150 million.
[16:38] flu it's designed to produce 150 million doses within six months of that.
[16:41] doses within six months of that decoration and so we were then going to.
[16:44] decoration and so we were then going to compete uh with that technology with RNA.
[16:47] compete uh with that technology with RNA was the.
[16:49] was the plan so this is state of the art in 2008.
[16:55] plan so this is state of the art in 2008 uh the invitro transcription reaction so.
[16:57] uh the invitro transcription reaction so for the first time we have no cells we.
[17:00] for the first time we have no cells we do not need a facility like that built.
[17:03] do not need a facility like that built in hly Springs and the key questions to.
[17:05] in hly Springs and the key questions to us were just what will the dose look.
[17:08] us were just what will the dose look like how is it going to be 100.
[17:10] like how is it going to be 100 micrograms one microgram or below the.
[17:13] micrograms one microgram or below the lower it is the small of the footprint.
[17:16] lower it is the small of the footprint and suddenly that facility can produce.
[17:18] and suddenly that facility can produce 28 different vaccines instead of just.
[17:22] 28 different vaccines instead of just one so you put you need a piece of DNA.
[17:25] One so you put you need a piece of DNA to run an initro transcription reaction.
[17:27] To run an initro transcription reaction you need a promoter sequence for the t7.
[17:30] You need a promoter sequence for the t7 polymerase and Inc comes the enzyme and.
[17:33] Polymerase and Inc comes the enzyme and it will pull in a sequence dependent.
[17:35] It will pull in a sequence dependent manner the nucleotide triphosphates and.
[17:38] Manner the nucleotide triphosphates and off comes your RNA in a sequence.
[17:40] Off comes your RNA in a sequence dependent Manner and it will just keep.
[17:42] Dependent Manner and it will just keep repeating and keep repeating um you.
[17:45] Repeating and keep repeating um you could buy these in commercial kits but.
[17:48] Could buy these in commercial kits but nobody had really scaled them back in uh.
[17:50] Nobody had really scaled them back in uh 2008 there was one little company in.
[17:53] 2008 there was one little company in Germany called covac who were pioneering.
[17:55] Germany called covac who were pioneering the space but they weren't really.
[17:57] The space but they weren't really publishing so we began B to explore.
[18:00] Publishing so we began B to explore scaling up this little trans uh this.
[18:03] Scaling up this little trans uh this invitro transcription reaction and.
[18:05] Invitro transcription reaction and here's our you know gel from about.
[18:10] Here's our you know gel from about 2009 um showing the ambient meas skip.
[18:14] 2009 um showing the ambient meas skip kit we were buying off the shelf this is.
[18:16] A 9kv RNA and here's our at scale uh kit.
[18:20] A 9kv RNA and here's our at scale uh kit independent system and the yields we're.
[18:23] independent system and the yields we're producing are around 5 milligrams per.
[18:27] producing are around 5 milligrams per mil this number this is this is not heard of in the protein world so already we're getting excited uh five migs per.
[18:35] M this is the process uh what it looks like when you buy one of those transcription reaction kits cap analoges back then weren't available so we're enzymatically capping and it's a batch process step by step so you run your RNA in vitro transcription reaction you then digest the DNA template lithium chloride precipitate at uh wash with ethanol and resuspend and dry um you then come in you denat the RNA you cap it more purification and done this is not an industrial scalable process and so we sat down and worked for about eight months to figure out how do we get this looking like an industrial process this is the original slide from.
[19:25] process this is the original slide from probably week one of the project um.
[19:28] probably week one of the project um nonviral delivery of self-amplifying RNA.
[19:32] nonviral delivery of self-amplifying RNA and this was our vision reinvent the gene vaccine uh develop a synthetic vaccine with the properties of a viral.
[19:33] and this was our vision reinvent the gene vaccine uh develop a synthetic vaccine with the properties of a viral vector uh without the complications of.
[19:37] vaccine with the properties of a viral vector uh without the complications of cell culture and anti vector immunity.
[19:39] vector uh without the complications of cell culture and anti vector immunity you see with the viral systems um uh that was safe scalable and widely.
[19:42] cell culture and anti vector immunity you see with the viral systems um uh that was safe scalable and widely applicable so we could drop in antigens.
[19:44] you see with the viral systems um uh that was safe scalable and widely applicable so we could drop in antigens from several different pathogens with the same process have several different.
[19:47] that was safe scalable and widely applicable so we could drop in antigens from several different pathogens with the same process have several different vaccines.
[19:50] applicable so we could drop in antigens from several different pathogens with the same process have several different vaccines.
[19:52] from several different pathogens with the same process have several different vaccines.
[19:54] the same process have several different vaccines.
[19:57] vaccines we were lucky enough to have uh a viral replicon particle uh system uh in place.
[19:59] we were lucky enough to have uh a viral replicon particle uh system uh in place.
[20:03] we were lucky enough to have uh a viral replicon particle uh system uh in place this is a packaging cell line that will take uh uh the same RNA we're using we're using self-amplifying RNA and use a packaging cell line to put a capsid around it an envelope and the glycoproteins it's based off alphav virus and so this is viral delivery we were trying to use a lipid nanoparticle.
[20:06] Replicon particle uh system uh in place this is a packaging cell line that will take uh uh the same RNA we're using we're using self-amplifying RNA and use a packaging cell line to put a capsid around it an envelope and the glycoproteins it's based off alphav virus and so this is viral delivery we were trying to use a lipid nanoparticle.
[20:08] this is a packaging cell line that will take uh uh the same RNA we're using we're using self-amplifying RNA and use a packaging cell line to put a capsid around it an envelope and the glycoproteins it's based off alphav virus and so this is viral delivery we were trying to use a lipid nanoparticle.
[20:11] take uh uh the same RNA we're using we're using self-amplifying RNA and use a packaging cell line to put a capsid around it an envelope and the glycoproteins it's based off alphav virus and so this is viral delivery we were trying to use a lipid nanoparticle.
[20:13] we're using self-amplifying RNA and use a packaging cell line to put a capsid around it an envelope and the glycoproteins it's based off alphav virus and so this is viral delivery we were trying to use a lipid nanoparticle.
[20:16] a packaging cell line to put a capsid around it an envelope and the glycoproteins it's based off alphav virus and so this is viral delivery we were trying to use a lipid nanoparticle.
[20:18] around it an envelope and the glycoproteins it's based off alphav virus and so this is viral delivery we were trying to use a lipid nanoparticle.
[20:20] glycoproteins it's based off alphav virus and so this is viral delivery we were trying to use a lipid nanoparticle.
[20:24] virus and so this is viral delivery we were trying to use a lipid nanoparticle.
[20:26] were trying to use a lipid nanoparticle system uh to deliver it and compare the
[20:28] system uh to deliver it and compare the two head-to-head in the clinic the
[20:31] two head-to-head in the clinic the alphav virus particle systems had been
[20:33] alphav virus particle systems had been used uh they worked well they were safe
[20:37] used uh they worked well they were safe efficacious and at about uh 10 to the 12
[20:40] efficacious and at about uh 10 to the 12 infectious units uh great data so we
[20:45] infectious units uh great data so we knew the RNA self-amplifying RNA from
[20:47] knew the RNA self-amplifying RNA from alpha viruses replicated in humans from
[20:50] alpha viruses replicated in humans from this system uh there were just a number
[20:52] this system uh there were just a number of questions could we even encapsulate
[20:54] of questions could we even encapsulate RNA in a lipid nanoparticle given the
[20:57] RNA in a lipid nanoparticle given the process uh would would it be stable
[20:59] process uh would would it be stable would it launch um were we using the
[21:02] would it launch um were we using the right capping strategies would we get
[21:04] right capping strategies would we get biological
[21:06] biological activity so this is the alpha virus uh
[21:09] activity so this is the alpha virus uh it's a family of uh over 45 species now
[21:15] it's a family of uh over 45 species now um this is the genome um at the five
[21:19] um this is the genome um at the five Prime end you have four non-structural
[21:21] Prime end you have four non-structural proteins that make the viral replicas uh
[21:24] proteins that make the viral replicas uh and allow amplification of the genome in
[21:28] and allow amplification of the genome in the midle here you have a subgenomic
[21:30] the midle here you have a subgenomic promoter and then at the three prime end
[21:33] promoter and then at the three prime end you have the capsid E1 and E2
[21:35] you have the capsid E1 and E2 glycoproteins so when I talked to you
[21:37] glycoproteins so when I talked to you about being able to make a
[21:39] about being able to make a multivalent vaccine we already know that
[21:42] multivalent vaccine we already know that the authentic Alpha virus is encoding
[21:45] the authentic Alpha virus is encoding this much we've certainly in
[21:46] this much we've certainly in synthetically been en able to encode
[21:49] synthetically been en able to encode more cargo than this and we're up to
[21:52] more cargo than this and we're up to five antigens now in
[21:54] five antigens now in 2020 it's positive sense so it rep
[21:58] 2020 it's positive sense so it rep replicates in the cytoplasm so we only
[22:00] replicates in the cytoplasm so we only need a delivery system to get us to the
[22:02] need a delivery system to get us to the cytoplasm with DNA cytoplasm through the
[22:06] cytoplasm with DNA cytoplasm through the nucleopore into the nucleus so from a
[22:08] nucleopore into the nucleus so from a delivery perspective it's a lot easier
[22:11] delivery perspective it's a lot easier and it's self amplifying
[22:15] and it's self amplifying self-replicating theoretically will need
[22:17] self-replicating theoretically will need a much lower
[22:19] a much lower dose so it looks just like a regular RNA
[22:23] dose so it looks just like a regular RNA shown up here five Prime cap polyat Tale
[22:26] shown up here five Prime cap polyat Tale utrs the only difference is you've got
[22:29] utrs the only difference is you've got these non-structural proteins and then
[22:31] these non-structural proteins and then you drop your antigen in after the
[22:33] you drop your antigen in after the subgenomic promoter that means with a
[22:37] subgenomic promoter that means with a single antigen you're already out to
[22:40] single antigen you're already out to 9,000 bases uh if you put five antigens
[22:44] 9,000 bases uh if you put five antigens in you're going to get close to 16,000
[22:47] in you're going to get close to 16,000 bases we've worked very hard at replica
[22:50] bases we've worked very hard at replica We Now can do a GMP compliant production
[22:53] We Now can do a GMP compliant production of a 16,000 base vector and it's taken
[22:57] of a 16,000 base vector and it's taken us many years of work to do
[23:03] that we don't use base modifications at
[23:07] that we don't use base modifications at all and so any of the all the work I'm
[23:10] all and so any of the all the work I'm going to show you today uh doesn't
[23:12] going to show you today uh doesn't contain a base modification it's wild
[23:14] contain a base modification it's wild type RNA um but you know it's through
[23:18] type RNA um but you know it's through covid and then after Nobel committee
[23:21] covid and then after Nobel committee awarded uh uh Drew wisman and Katy kuo
[23:25] awarded uh uh Drew wisman and Katy kuo uh a prized for their pivotal work in
[23:27] uh a prized for their pivotal work in the field we certainly explored it but
[23:29] the field we certainly explored it but for vaccine applications it didn't work
[23:32] for vaccine applications it didn't work and what they're doing is moving from
[23:35] and what they're doing is moving from iDine to a Pudo urine or methy Pudu
[23:38] iDine to a Pudo urine or methy Pudu iDine analog and that's what we saw
[23:40] iDine analog and that's what we saw going forward in the covid uh vaccine
[23:43] going forward in the covid uh vaccine trials uh it allows you to dose higher
[23:45] trials uh it allows you to dose higher doses um as I'll show you as we move
[23:49] doses um as I'll show you as we move forward with these types of replicating
[23:52] forward with these types of replicating vaccines the doses are much
[23:55] vaccines the doses are much lower so now there are five five
[23:58] lower so now there are five five different types of RNA back in 2008 we
[24:02] different types of RNA back in 2008 we had one company curvac in Germany with
[24:05] had one company curvac in Germany with unmodified RNA there's us with self uh
[24:09] unmodified RNA there's us with self uh amplifying it was called then the Sam
[24:11] amplifying it was called then the Sam vaccine platform uh since then base
[24:14] vaccine platform uh since then base modified trans amplifying circular RNA
[24:18] modified trans amplifying circular RNA I'm sure every year there'll be a
[24:19] I'm sure every year there'll be a different type of RNA appearing uh but
[24:22] different type of RNA appearing uh but they all have the same principle
[24:24] they all have the same principle enzimatic transcription figure out
[24:27] enzimatic transcription figure out delivery we move forward very
[24:30] delivery we move forward very fast so how does an alpha virus work
[24:33] fast so how does an alpha virus work we'll start with linear RNA first so
[24:36] we'll start with linear RNA first so here's your linear RNA we get it into
[24:38] here's your linear RNA we get it into the cytoplasm it engages the ribosome uh
[24:41] the cytoplasm it engages the ribosome uh it's encoding our antigen and then then
[24:43] it's encoding our antigen and then then it'll make a small amount of the antigen
[24:46] it'll make a small amount of the antigen usually over a period of 24 hours
[24:50] usually over a period of 24 hours self-amplifying systems once inside the
[24:52] self-amplifying systems once inside the cell
[24:55] cell they replicate in what's known as a
[24:57] they replicate in what's known as a spher
[24:58] spher it's a membrane privileged BB and it's
[25:01] it's a membrane privileged BB and it's able to replicate for several days the
[25:05] able to replicate for several days the first thing it'll do is make a negative
[25:07] first thing it'll do is make a negative sense copy of its template and off that
[25:09] sense copy of its template and off that it amplifies The genome but it also
[25:12] it amplifies The genome but it also launches the sub genome this looks like
[25:14] launches the sub genome this looks like endogenous RNA within the cell it binds
[25:17] endogenous RNA within the cell it binds the ribosome and starts to make the
[25:19] the ribosome and starts to make the structural proteins the capsid will pick
[25:22] structural proteins the capsid will pick up the genome the glycoproteins are on
[25:26] up the genome the glycoproteins are on the envelope and it will Bud off live
[25:28] the envelope and it will Bud off live infectious virus because we've deleted
[25:32] infectious virus because we've deleted the structural proteins we can't do this
[25:34] the structural proteins we can't do this but we do replicate in the BLB we make
[25:38] but we do replicate in the BLB we make messenger RNA but encoding our antigen
[25:41] messenger RNA but encoding our antigen and I think all the examples are going
[25:43] and I think all the examples are going to show you those are glycoproteins we
[25:45] to show you those are glycoproteins we put on the cell surface so it's deadend
[25:48] put on the cell surface so it's deadend delivery but it's mimicking of viral
[25:50] delivery but it's mimicking of viral infection so to the cell and
[25:53] infection so to the cell and cells that are transfected with material
[25:57] cells that are transfected with material they are
[25:58] they are in danger signal mode they're seeing
[26:01] in danger signal mode they're seeing that they're starting to the RNA is
[26:03] that they're starting to the RNA is amplifying and the cell intracellular
[26:05] amplifying and the cell intracellular helicases at some point are going to
[26:07] helicases at some point are going to detect it and say we've there's a viral
[26:10] detect it and say we've there's a viral infection going off somewhere the lipid
[26:13] infection going off somewhere the lipid nanoparticle will traffic into the
[26:15] nanoparticle will traffic into the draining lymph node within hours and be
[26:18] draining lymph node within hours and be picked up by the immune system you get
[26:20] picked up by the immune system you get an innate immune response to that
[26:23] an innate immune response to that because it looks like a virus with RNA
[26:25] because it looks like a virus with RNA inside and with that combination of
[26:29] inside and with that combination of events it looks very much like a live
[26:32] events it looks very much like a live attenuated vaccine so it's very potent
[26:35] attenuated vaccine so it's very potent we get strong antibody strong te- cell
[26:40] responses half of my brain I'm an
[26:43] responses half of my brain I'm an engineer and so this is how I think
[26:45] engineer and so this is how I think these are the the pillars of vaccine
[26:47] these are the the pillars of vaccine design in my world uh and the antigen is
[26:51] design in my world uh and the antigen is the most overlooked content so I always
[26:53] the most overlooked content so I always get excited when I'm in places where
[26:55] get excited when I'm in places where they're designing and discovering new
[26:57] they're designing and discovering new antigens
[26:59] antigens because I can design new vectors
[27:01] because I can design new vectors delivery system and Manufacturing if we
[27:04] delivery system and Manufacturing if we want new vaccines we need new antigens
[27:07] want new vaccines we need new antigens and so when I come in a place like this
[27:08] and so when I come in a place like this I get very excited about possibilities
[27:11] I get very excited about possibilities of uh making new uh vaccines and getting
[27:14] of uh making new uh vaccines and getting them into the the public and and saving
[27:17] them into the the public and and saving lives so you need a very well-designed
[27:20] lives so you need a very well-designed antigen I'm going to talk to you about
[27:22] antigen I'm going to talk to you about self-amplifying self-replicating RNA
[27:25] self-amplifying self-replicating RNA lipid nanoparticle Delivery Systems uh
[27:28] lipid nanoparticle Delivery Systems uh and you know manufacturer of those so
[27:32] and you know manufacturer of those so we'll start with RNA uh synthesis using
[27:35] we'll start with RNA uh synthesis using the ivt reaction so that transcription
[27:39] the ivt reaction so that transcription reaction kit that schematic I showed has
[27:41] reaction kit that schematic I showed has got a change if we're going to go to
[27:43] got a change if we're going to go to commercial scale commercial scale for
[27:46] commercial scale commercial scale for self-replicating RNA that I work on wear
[27:49] self-replicating RNA that I work on wear less than a liter bio reactor uh
[27:51] less than a liter bio reactor uh probably about a half a liter so it's
[27:54] probably about a half a liter so it's very small very compact madna bisa biion
[27:58] very small very compact madna bisa biion Tech they're at about a 30 liter scale
[28:00] Tech they're at about a 30 liter scale so quite a large vessel um but here's
[28:03] so quite a large vessel um but here's your bioreactor everything has to happen
[28:05] your bioreactor everything has to happen in there so you can't do stepwise
[28:08] in there so you can't do stepwise production intermediate purifications so
[28:11] production intermediate purifications so we had to quickly figure out how to do a
[28:13] we had to quickly figure out how to do a onepot RNA synthesis so we you make the
[28:17] onepot RNA synthesis so we you make the RNA you did digest the dnas and then you
[28:21] RNA you did digest the dnas and then you you come in and you cap it all in the
[28:23] you come in and you cap it all in the one pot no intermediates just one two
[28:26] one pot no intermediates just one two three steps and then into to
[28:28] three steps and then into to purification certainly in 2008 there
[28:31] purification certainly in 2008 there really wasn't much available uh now
[28:34] really wasn't much available uh now through covid there's a lot of
[28:36] through covid there's a lot of purification Technologies available
[28:39] purification Technologies available built for purpose ready for RNA so you
[28:42] built for purpose ready for RNA so you have to purify away from the fragmented
[28:46] have to purify away from the fragmented DNA the ntps the enzymes um any
[28:51] DNA the ntps the enzymes um any antibiotic that may have come through
[28:54] antibiotic that may have come through during ecoli production of the
[28:56] during ecoli production of the plasmid but the purification levels that
[29:00] plasmid but the purification levels that you need to achieve are monumentally
[29:04] you need to achieve are monumentally smaller than anything from cell culture
[29:06] smaller than anything from cell culture we don't have any of the debris coming
[29:08] we don't have any of the debris coming through from from that hostel production
[29:11] through from from that hostel production line so it it's truly a lot easier right
[29:14] line so it it's truly a lot easier right from the beginning in 2008 we were
[29:16] from the beginning in 2008 we were looking at the residual content of the
[29:19] looking at the residual content of the raw reaction mixture and we were pretty
[29:22] raw reaction mixture and we were pretty close to a an ecoli produced uh vaccine
[29:26] close to a an ecoli produced uh vaccine without any purifications so there
[29:28] without any purifications so there wasn't much that needed to happen but we
[29:31] wasn't much that needed to happen but we had to get creative and figure out which
[29:33] had to get creative and figure out which contaminants were inhibiting uh
[29:38] immunogenicity we need to dialy and
[29:40] immunogenicity we need to dialy and concentrate at an industrial scale
[29:42] concentrate at an industrial scale that's tangental flow filtration I've
[29:44] that's tangental flow filtration I've been it up in the labs and I've seen
[29:46] been it up in the labs and I've seen systems there so you know all about it
[29:48] systems there so you know all about it uh and then
[29:50] uh and then sterilization um yields even at scale we
[29:53] sterilization um yields even at scale we up to close to 8 milligram per mil so
[29:57] up to close to 8 milligram per mil so moving from the kit we were able to
[29:58] moving from the kit we were able to scale quite well and keep production
[30:01] scale quite well and keep production high and we found that this is a generic
[30:05] high and we found that this is a generic process small tweaks to the the process
[30:08] process small tweaks to the the process when we change antigens change vectors
[30:11] when we change antigens change vectors uh but it's pretty
[30:13] uh but it's pretty generic so LMP uh delivery so there's a
[30:18] generic so LMP uh delivery so there's a lot of this is the first publication of
[30:20] lot of this is the first publication of an LMP delivered RNA vaccine back in
[30:23] an LMP delivered RNA vaccine back in 2012 this work was actually done in 2008
[30:27] 2012 this work was actually done in 2008 uh but we had to work hard with the
[30:28] uh but we had to work hard with the patent lawyers to file patents uh but
[30:31] patent lawyers to file patents uh but we're encapsulating the RNA
[30:33] we're encapsulating the RNA encapsulation efficiencies were very
[30:35] encapsulation efficiencies were very high even with the first experiments uh
[30:38] high even with the first experiments uh these long viral derived RNA uh sit when
[30:43] these long viral derived RNA uh sit when we looked by uh cryo em and other
[30:46] we looked by uh cryo em and other techniques we're
[30:47] techniques we're about 60 nanometers in a ball uh
[30:51] about 60 nanometers in a ball uh phosphate backbone on the outside bases
[30:53] phosphate backbone on the outside bases on the middle just ideal for
[30:56] on the middle just ideal for encapsulating a lot EAS easier than
[30:58] encapsulating a lot EAS easier than linear RNA in most
[31:00] linear RNA in most respects um in our crude first
[31:03] respects um in our crude first experiments we were about 140 nimet now
[31:06] experiments we were about 140 nimet now we're down close to the size of a virus
[31:09] we're down close to the size of a virus which is at about 70 80
[31:12] which is at about 70 80 nanm composed of four components
[31:14] nanm composed of four components phospholipid cholesterol that the base
[31:17] phospholipid cholesterol that the base stabilizing uh the the the lipid
[31:20] stabilizing uh the the the lipid envelope um and a peg lipid on the
[31:23] envelope um and a peg lipid on the outside we we needed the peg lipid in
[31:26] outside we we needed the peg lipid in order to be able to man manufacture the
[31:28] order to be able to man manufacture the LMP we've got to process it Downstream
[31:32] LMP we've got to process it Downstream through tangental flow filtration
[31:33] through tangental flow filtration sterile filtration without the peg it's
[31:36] sterile filtration without the peg it's too sticky and we can't process it um
[31:40] too sticky and we can't process it um the most important was this ionizable
[31:42] the most important was this ionizable catonic lipid and we'll come to it in
[31:43] catonic lipid and we'll come to it in the subsequent slides um I'll just say
[31:47] the subsequent slides um I'll just say that for us uh we had to publish a lot a
[31:50] that for us uh we had to publish a lot a lot of the early team were posts uh uh
[31:54] lot of the early team were posts uh uh we had a few PhD students later on so we
[31:57] we had a few PhD students later on so we pulled this ionizable catonic lipid from
[31:59] pulled this ionizable catonic lipid from the literature from petsis group it had
[32:02] the literature from petsis group it had been published we had a big pot
[32:04] been published we had a big pot synthesized and for everything we
[32:06] synthesized and for everything we published this is the lipid we used but
[32:08] published this is the lipid we used but internally we had stuff that was a lot
[32:11] internally we had stuff that was a lot more potent um but this worked pretty
[32:14] more potent um but this worked pretty well even from the
[32:16] well even from the beginning so this is what the process
[32:18] beginning so this is what the process looked like this is our first prototype
[32:21] looked like this is our first prototype it's a Gerry rigged uh syringe driver um
[32:24] it's a Gerry rigged uh syringe driver um we've got a one syringe filled with the
[32:27] we've got a one syringe filled with the lip in ethanol the RNA is in buffer
[32:30] lip in ethanol the RNA is in buffer we've got a dilution buffer and we're
[32:33] we've got a dilution buffer and we're shooting it through a t- mixer um pH is
[32:38] shooting it through a t- mixer um pH is low ph6 so that lipid is protonated uh
[32:42] low ph6 so that lipid is protonated uh it it's uh positively charged the
[32:47] it it's uh positively charged the phosphate backbone's negatively charged
[32:49] phosphate backbone's negatively charged so the two lipids interact and then the
[32:51] so the two lipids interact and then the rest of the lipids Pile in and you form
[32:53] rest of the lipids Pile in and you form your
[32:55] your lipm we're at 33% ethanol it's metast
[32:59] lipm we're at 33% ethanol it's metast stable and you hold it for an hour
[33:01] stable and you hold it for an hour everything stabilizes the particles
[33:04] everything stabilizes the particles start to rearrange and then finally you
[33:07] start to rearrange and then finally you bring up the pH by diluting with a
[33:09] bring up the pH by diluting with a phosphate buffer saline uh and then
[33:11] phosphate buffer saline uh and then you're into tangental flow filtration to
[33:14] you're into tangental flow filtration to diiz away the ethanol and change the
[33:17] diiz away the ethanol and change the buffer into your final formulation
[33:18] buffer into your final formulation buffer now because I'm a nartist we're
[33:22] buffer now because I'm a nartist we're all about scalability we don't want to
[33:24] all about scalability we don't want to do spin columns or artificial dialysis
[33:27] do spin columns or artificial dialysis bags we were lucky enough that Spectrum
[33:30] bags we were lucky enough that Spectrum Labs had just launched this new
[33:33] Labs had just launched this new crosso uh tff system and at a 2 mil
[33:36] crosso uh tff system and at a 2 mil volume we could process this LMP and so
[33:40] volume we could process this LMP and so we could mimic a process at 2 Ms that we
[33:44] we could mimic a process at 2 Ms that we could do at 100 lers and you can connect
[33:47] could do at 100 lers and you can connect it up and measure trans member and
[33:49] it up and measure trans member and pressures and everything a chemical
[33:51] pressures and everything a chemical engineer wants to know about what
[33:53] engineer wants to know about what happens so through as I scale and so at
[33:56] happens so through as I scale and so at a 2 mil scale we could capture all the
[33:59] a 2 mil scale we could capture all the information and do predictions on
[34:03] information and do predictions on scalability uh we then filter it 02
[34:06] scalability uh we then filter it 02 micron filter from from a pharmaceutics
[34:09] micron filter from from a pharmaceutics perspective you need to be under 100
[34:12] perspective you need to be under 100 nanometers in order to be not blocking
[34:14] nanometers in order to be not blocking that filter if you look at the
[34:16] that filter if you look at the distribution of your Dynamic like
[34:18] distribution of your Dynamic like scattering if you're anything above 200
[34:22] scattering if you're anything above 200 nanometers is blocking that filter So
[34:24] nanometers is blocking that filter So you you're running
[34:26] you you're running a volume through the filter versus time
[34:29] a volume through the filter versus time eventually the back pressure is buil and
[34:31] eventually the back pressure is buil and it will
[34:32] it will block so this is what a process looks
[34:35] block so this is what a process looks like now you have a mixing device and
[34:38] like now you have a mixing device and we'll come on to you know my favorite
[34:40] we'll come on to you know my favorite mixing device it's just one example of
[34:43] mixing device it's just one example of of
[34:44] of of but it was a severe problem back in
[34:47] of but it was a severe problem back in 2008 that needed to be solved we were
[34:50] 2008 that needed to be solved we were not moving forward with that syringe
[34:52] not moving forward with that syringe driver t- mixing setup uh we felt
[34:55] driver t- mixing setup uh we felt tangental flow filtration sterile
[34:57] tangental flow filtration sterile filtration was saved were were were safe
[35:01] filtration was saved were were were safe it's standard operating equipment in
[35:03] it's standard operating equipment in most um uh Labs at industry scale uh but
[35:07] most um uh Labs at industry scale uh but the membranes certainly weren't uh where
[35:10] the membranes certainly weren't uh where they needed to be since covid uh we've
[35:13] they needed to be since covid uh we've seen prototype membranes being developed
[35:16] seen prototype membranes being developed specifically for lipid nanoparticles and
[35:18] specifically for lipid nanoparticles and RNA and that's a good thing for the
[35:20] RNA and that's a good thing for the field so here we go early proof of
[35:23] field so here we go early proof of concept uh it's all published in these
[35:25] concept uh it's all published in these two papers there are several others
[35:27] two papers there are several others these are just my two favorite uh the
[35:30] these are just my two favorite uh the first one uh is we're tackling a
[35:32] first one uh is we're tackling a respiratory sensial virus um it's
[35:36] respiratory sensial virus um it's uh an envelope viran the key Target is
[35:40] uh an envelope viran the key Target is this rsbf protein internally at nus at
[35:44] this rsbf protein internally at nus at the time there was a subunit protein
[35:47] the time there was a subunit protein vaccine being developed so we had all of
[35:50] vaccine being developed so we had all of the um protein Design coming from them
[35:53] the um protein Design coming from them that we knew it needed to be a trer they
[35:56] that we knew it needed to be a trer they were doing Fusion deletion uh approaches
[35:58] were doing Fusion deletion uh approaches and all of that information was
[36:00] and all of that information was available to us to drop in to our RNA
[36:04] available to us to drop in to our RNA vaccine however it's a competition
[36:07] vaccine however it's a competition whoever is the best goes forward into
[36:09] whoever is the best goes forward into the clinic and we to in order for us to
[36:13] the clinic and we to in order for us to have been chosen we would have to be
[36:15] have been chosen we would have to be superior equivalence isn't going to
[36:17] superior equivalence isn't going to count there is no manufacturing
[36:19] count there is no manufacturing infrastructure for RNA making a subunit
[36:22] infrastructure for RNA making a subunit agented with alum is big farmers bread
[36:25] agented with alum is big farmers bread and butter so but this was the first
[36:27] and butter so but this was the first Target not my choice but it's a
[36:31] Target not my choice but it's a competitive environment in big
[36:33] competitive environment in big farmer so here's the first experiment
[36:36] farmer so here's the first experiment we're in BC mice we're vaccinating twice
[36:39] we're in BC mice we're vaccinating twice three weeks apart taking a bleed either
[36:41] three weeks apart taking a bleed either post first at day 14 or postc at day 35
[36:45] post first at day 14 or postc at day 35 we're just measuring antibodies by Eliza
[36:49] we're just measuring antibodies by Eliza specific to that F protein we have naked
[36:52] specific to that F protein we have naked RNA here this is post first so naked RNA
[36:55] RNA here this is post first so naked RNA LMP formulated 10 micrograms down to uh
[37:00] LMP formulated 10 micrograms down to uh 01 microgram this is the control so this
[37:04] 01 microgram this is the control so this is this virus viral Replicon particle so
[37:08] is this virus viral Replicon particle so it's the same RNA but we're delivering
[37:10] it's the same RNA but we're delivering it virally um it's probably a picogram
[37:13] it virally um it's probably a picogram of RNA it's pretty low uh one to the six
[37:17] of RNA it's pretty low uh one to the six infectious units this is at the upper
[37:19] infectious units this is at the upper level of the dose response so this is a
[37:22] level of the dose response so this is a good control if we can match it uh we're
[37:25] good control if we can match it uh we're in good shape and we know that this
[37:26] in good shape and we know that this works clinically Inhumans the vrp and so
[37:30] works clinically Inhumans the vrp and so what you can see post first we we we
[37:33] what you can see post first we we we were utterly amazed all the mice Zer
[37:35] were utterly amazed all the mice Zer converted and we saw a dose response
[37:37] converted and we saw a dose response it's very flat because it's
[37:38] it's very flat because it's self-amplifying system but all the my
[37:41] self-amplifying system but all the my Ser converted we then boosted them and
[37:44] Ser converted we then boosted them and they all boosted and at one to 0.1
[37:47] they all boosted and at one to 0.1 microgram were equivalent to this
[37:50] microgram were equivalent to this vrp and so we're very excited go read
[37:54] vrp and so we're very excited go read the paper there's lots of t- cell
[37:55] the paper there's lots of t- cell responses but for RSV it's all about
[37:58] responses but for RSV it's all about antibodies and neutralizing antibodies
[38:00] antibodies and neutralizing antibodies so we went into head-to-head competition
[38:03] so we went into head-to-head competition with the subunit now we're in the cotton
[38:06] with the subunit now we're in the cotton wrap model where you can do an
[38:07] wrap model where you can do an intranasal challenge very similar setup
[38:10] intranasal challenge very similar setup two doses uh three weeks apart we're
[38:13] two doses uh three weeks apart we're going to look at some serology at day 35
[38:16] going to look at some serology at day 35 and then look at viral tits in the lung
[38:19] and then look at viral tits in the lung after challenge uh PBS as a control
[38:23] after challenge uh PBS as a control naked RNA is another control one
[38:26] naked RNA is another control one microgram of RNA encapsulated in our LMP
[38:29] microgram of RNA encapsulated in our LMP the viral control system at 10 to the
[38:32] the viral control system at 10 to the six and then the Alum agented subunit uh
[38:38] six and then the Alum agented subunit uh at 10
[38:39] at 10 micrograms and what you can see here on
[38:41] micrograms and what you can see here on the new tius we're not at the levels of
[38:44] the new tius we're not at the levels of the subunit uh we're equivalent to the
[38:46] the subunit uh we're equivalent to the vrp um looking at uh antibodies by Liza
[38:51] vrp um looking at uh antibodies by Liza again we're we're below uh but we're
[38:53] again we're we're below uh but we're seeing good levels of protection uh
[38:56] seeing good levels of protection uh really no different between any of the
[38:59] really no different between any of the vaccines based on this data and other
[39:01] vaccines based on this data and other datas there's no path forward uh for the
[39:05] datas there's no path forward uh for the first iterations of our RNA technology
[39:09] first iterations of our RNA technology uh That vaccine the subunit vaccine is
[39:13] uh That vaccine the subunit vaccine is the one that got commercialized by uh uh
[39:15] the one that got commercialized by uh uh GSK recently so it's very effective but
[39:19] GSK recently so it's very effective but in order to win you've got to be
[39:21] in order to win you've got to be competitive
[39:23] competitive so in order to be competitive we needed
[39:26] so in order to be competitive we needed money we needed innovate so I reached
[39:29] money we needed innovate so I reached out uh through uh my networks at uh
[39:33] out uh through uh my networks at uh novatis I'd been told no so many times
[39:37] novatis I'd been told no so many times uh there's no more money in the budget
[39:39] uh there's no more money in the budget finally the CEO told me you can go get a
[39:42] finally the CEO told me you can go get a grant and and so I went out and got a
[39:44] grant and and so I went out and got a very big grun uh from uh the defense
[39:47] very big grun uh from uh the defense Advanced research project agency uh it
[39:50] Advanced research project agency uh it it's valued there $14.2 million but it's
[39:54] it's valued there $14.2 million but it's a cost sharing initiative so as a senior
[39:56] a cost sharing initiative so as a senior research science ITI at nartis suddenly
[39:59] research science ITI at nartis suddenly I have $28.4 million to spend to
[40:02] I have $28.4 million to spend to innovate on the program people were kind
[40:04] innovate on the program people were kind of sleeping during the writing of the
[40:06] of sleeping during the writing of the project plans not really paying much
[40:10] project plans not really paying much attention because I don't think they
[40:11] attention because I don't think they thought we were going to get it and so
[40:13] thought we were going to get it and so what I wrote in there was we wanted to
[40:15] what I wrote in there was we wanted to go after disease targets that there were
[40:18] go after disease targets that there were commercial vaccines against there were
[40:20] commercial vaccines against there were Coral of protection and so we started to
[40:22] Coral of protection and so we started to do studies in this contract on flu and
[40:28] do studies in this contract on flu and rabies and so which was a little
[40:31] rabies and so which was a little controversial because we didn't need a a
[40:33] controversial because we didn't need a a new flu vaccine because we had the new
[40:36] new flu vaccine because we had the new uh uh facility B built in hly Springs uh
[40:40] uh uh facility B built in hly Springs uh but it was sold by me as mechanism of
[40:43] but it was sold by me as mechanism of action understanding of the technology
[40:46] action understanding of the technology we'll understand it we'll improve it and
[40:48] we'll understand it we'll improve it and then we'll find where we'll fit
[40:51] then we'll find where we'll fit it oh and and so we were not alone in in
[40:55] it oh and and so we were not alone in in in DARPA funding uh CAC sopi incel art
[40:59] in DARPA funding uh CAC sopi incel art madna they nartis pumped over a billion
[41:02] madna they nartis pumped over a billion dollars into the field in and around
[41:06] dollars into the field in and around 2012 and because of that money The
[41:09] 2012 and because of that money The Innovation happened the data started to
[41:12] Innovation happened the data started to look much better Venture Capital started
[41:15] look much better Venture Capital started spending the banks started spending a
[41:17] spending the banks started spending a big farmer got interested so I can't
[41:20] big farmer got interested so I can't sing enough praises to Dara uh they get
[41:23] sing enough praises to Dara uh they get no recognition for it but we certainly
[41:26] no recognition for it but we certainly have have seen benefit for
[41:28] have have seen benefit for it
[41:30] it so the next Target is flu I think
[41:33] so the next Target is flu I think everyone knows here flu the major
[41:35] everyone knows here flu the major antigen of neutralization is hemog
[41:38] antigen of neutralization is hemog glutenin uh so we're now just we've
[41:40] glutenin uh so we're now just we've switched from RSV F now to hemog
[41:43] switched from RSV F now to hemog glutenin and dropping it into our
[41:45] glutenin and dropping it into our self-amplifying RNA here's the first
[41:47] self-amplifying RNA here's the first experiment it's H1 from uh uh the a
[41:51] experiment it's H1 from uh uh the a California swine flu strain from
[41:54] California swine flu strain from 2009 we are uh measuring hemog gluten
[41:58] 2009 we are uh measuring hemog gluten inefficient titis so functional
[42:00] inefficient titis so functional neutralizing antibodies uh the target
[42:03] neutralizing antibodies uh the target again the the the the benchmarks are the
[42:08] again the the the the benchmarks are the material coming from that hly Springs
[42:10] material coming from that hly Springs site either with or without an adant at
[42:13] site either with or without an adant at the upper dose uh point in the
[42:16] the upper dose uh point in the mouse we're dosing uh at 0.1 and one
[42:19] mouse we're dosing uh at 0.1 and one micrograms in an LMP we're either doing
[42:22] micrograms in an LMP we're either doing a 3-we interval between vaccinations or
[42:26] a 3-we interval between vaccinations or uh uh an 8we interval the longer the
[42:29] uh uh an 8we interval the longer the interval between the vaccination the
[42:32] interval between the vaccination the stronger the boosting response with
[42:33] stronger the boosting response with these self-amplifying systems uh and
[42:36] these self-amplifying systems uh and this was one of the first experiments to
[42:37] this was one of the first experiments to do that if you go to 16 weeks the Titus
[42:40] do that if you go to 16 weeks the Titus go even higher so at three weeks what
[42:43] go even higher so at three weeks what we're seeing is we're seeing uh priming
[42:46] we're seeing is we're seeing uh priming uh events happening they're equivalent
[42:49] uh events happening they're equivalent to the the subunit without an adant but
[42:52] to the the subunit without an adant but we're not at the levels with mf59 I
[42:54] we're not at the levels with mf59 I think everyone's familiar with mf59 here
[42:57] think everyone's familiar with mf59 here so it's a high bar in the flu
[42:59] so it's a high bar in the flu world we boost our responses boost all
[43:03] world we boost our responses boost all Mouse have zeroc converted in that LMP
[43:06] Mouse have zeroc converted in that LMP formulation as we've seen before the
[43:08] formulation as we've seen before the data's clustering nicely uh we're at the
[43:10] data's clustering nicely uh we're at the level of the Su unit at 1 microgram but
[43:13] level of the Su unit at 1 microgram but here's
[43:14] here's mf59 we increased the interval of the
[43:18] mf59 we increased the interval of the the Boost and now we're starting to
[43:21] the Boost and now we're starting to differentiate for the unagented but
[43:23] differentiate for the unagented but we're still not at the levels of the
[43:25] we're still not at the levels of the agented vaccines but but at least we're
[43:27] agented vaccines but but at least we're on Target and we began several years of
[43:32] on Target and we began several years of optimization of the vector of the
[43:34] optimization of the vector of the delivery system pushing the dose down
[43:37] delivery system pushing the dose down and pushing potency up none of that data
[43:39] and pushing potency up none of that data uh has been
[43:41] uh has been published but one of the true advantages
[43:45] published but one of the true advantages of RNA was speed this is the first
[43:49] of RNA was speed this is the first example published in the literature of
[43:51] example published in the literature of speed of the technology so in
[43:54] speed of the technology so in 201 uh 13 in March Mar three people in
[43:58] 201 uh 13 in March Mar three people in China died of uh a novel bird flu an
[44:03] China died of uh a novel bird flu an h7n9 the Chinese CDC put the sequence
[44:06] h7n9 the Chinese CDC put the sequence online on a global sharing database in
[44:10] online on a global sharing database in the paper uh you'll find two members of
[44:14] the paper uh you'll find two members of the Chinese CDC as co-authors as the
[44:16] the Chinese CDC as co-authors as the thank you for sharing data allowing us
[44:19] thank you for sharing data allowing us uh to do Global surveillance of of flu
[44:23] uh to do Global surveillance of of flu but we were collaborating with Craig
[44:26] but we were collaborating with Craig Venter in in San Diego uh they just
[44:29] Venter in in San Diego uh they just invented Gibson synthesis so they were
[44:31] invented Gibson synthesis so they were able to quickly synthesize the ha Gene
[44:34] able to quickly synthesize the ha Gene in San Diego um proof freed it check it
[44:38] in San Diego um proof freed it check it and then by day three uh it it's it
[44:41] and then by day three uh it it's it arrived um in in Boston into my facility
[44:46] arrived um in in Boston into my facility uh we cloned it into our plasmid we grew
[44:48] uh we cloned it into our plasmid we grew up the plasmid we made RNA um uh and so
[44:52] up the plasmid we made RNA um uh and so by day eight we're showing here on an
[44:55] by day eight we're showing here on an agos gel here's are we're calling them
[44:58] agos gel here's are we're calling them Sam vaccine self-amplifying message the
[45:00] Sam vaccine self-amplifying message the marketers by this point have got a hold
[45:02] marketers by this point have got a hold of us we're not calling this
[45:04] of us we're not calling this self-amplifying RNA we're not calling it
[45:06] self-amplifying RNA we're not calling it a replica we need a a name that's more
[45:09] a replica we need a a name that's more friendly to the public and so we're now
[45:11] friendly to the public and so we're now the Sam vaccine platform but here it is
[45:14] the Sam vaccine platform but here it is around 10kb containing that novel 87
[45:18] around 10kb containing that novel 87 there's a sequence online there are no
[45:21] there's a sequence online there are no viral isolates in the US at all uh we
[45:24] viral isolates in the US at all uh we then throw it on a uh on sales
[45:27] then throw it on a uh on sales uh and we look on the cell surface of
[45:29] uh and we look on the cell surface of the the cells and run a western blot and
[45:33] the the cells and run a western blot and here's the H7 we had an antibody from an
[45:36] here's the H7 we had an antibody from an earlier an 87 N3 that crossreacted and
[45:40] earlier an 87 N3 that crossreacted and so we've got full length RNA it's
[45:43] so we've got full length RNA it's replicating in cell culture and it's
[45:45] replicating in cell culture and it's producing the antigen we repurposed some
[45:49] producing the antigen we repurposed some Mice from a an experiment and so by day
[45:51] Mice from a an experiment and so by day 13 we vaccinated mice either at a 3-we
[45:55] 13 we vaccinated mice either at a 3-we interval or an 8 we interval there are
[45:57] interval or an 8 we interval there are no controls available there's no there's
[46:00] no controls available there's no there's no subunit vaccine existing um this is
[46:04] no subunit vaccine existing um this is the first pandemic response uh uh
[46:08] the first pandemic response uh uh documented you can go read about it in
[46:10] documented you can go read about it in in this paper we're measuring hog
[46:13] in this paper we're measuring hog glutenin inhibition at uh Titus naked
[46:17] glutenin inhibition at uh Titus naked RNA formulated one microgram we have a
[46:20] RNA formulated one microgram we have a negative control an H1 uh uh ha I showed
[46:25] negative control an H1 uh uh ha I showed you previously we shouldn't see
[46:27] you previously we shouldn't see conversion so we're priming we're seeing
[46:30] conversion so we're priming we're seeing responses we're boosting uh at week
[46:33] responses we're boosting uh at week three 3 we interval at uh day 35 we're
[46:37] three 3 we interval at uh day 35 we're above 40 the coral of protection uh so
[46:40] above 40 the coral of protection uh so the vaccines working well very well if
[46:42] the vaccines working well very well if we boost at an 8we interval now we have
[46:44] we boost at an 8we interval now we have titter up in the thousands so now we're
[46:47] titter up in the thousands so now we're starting to be competitive we have speed
[46:50] starting to be competitive we have speed is an advantage for the
[46:53] is an advantage for the technology this is the last paper that I
[46:55] technology this is the last paper that I wrote while I was at uh of artist I
[46:57] wrote while I was at uh of artist I wrote it with Jeff ommer who I showed
[46:59] wrote it with Jeff ommer who I showed you that picture of uh it was called
[47:01] you that picture of uh it was called vaccines on demand science fiction or
[47:04] vaccines on demand science fiction or future reality here's a statement it's
[47:07] future reality here's a statement it's basically saying RNA vaccines are the
[47:09] basically saying RNA vaccines are the future for pandemic uh and we started to
[47:12] future for pandemic uh and we started to argue about uh why that would be and
[47:15] argue about uh why that would be and production capacity and so here is a a
[47:18] production capacity and so here is a a graph uh we're looking at a human doses
[47:21] graph uh we're looking at a human doses per liter um and as the dose comes down
[47:25] per liter um and as the dose comes down if we can get to a human dose of 1
[47:27] if we can get to a human dose of 1 microgram we'll be able to produce 5
[47:31] microgram we'll be able to produce 5 million human doses per liter in a few
[47:33] million human doses per liter in a few hours and so the the site and Holly
[47:36] hours and so the the site and Holly Springs are now getting very worried I
[47:38] Springs are now getting very worried I was thrown out of that building given a
[47:40] was thrown out of that building given a seminar I told I wasn't welcome because
[47:43] seminar I told I wasn't welcome because they were going to have to eventually
[47:44] they were going to have to eventually Embrace a new technology and change and
[47:47] Embrace a new technology and change and change is always very difficult we
[47:49] change is always very difficult we calculated the viral delivery uh system
[47:52] calculated the viral delivery uh system that Viral Replicon particle in humans
[47:55] that Viral Replicon particle in humans 10 to the8 was the dose uh it was about
[47:58] 10 to the8 was the dose uh it was about 0.5 nanograms so from a scientific
[48:01] 0.5 nanograms so from a scientific perspective we thought one or 01
[48:05] perspective we thought one or 01 microgram if you got everything right
[48:07] microgram if you got everything right was probably doable we're never going to
[48:09] was probably doable we're never going to be as efficient during delivery as a
[48:11] be as efficient during delivery as a virus with an LMP but maybe if we
[48:14] virus with an LMP but maybe if we optimize them we can get within you know
[48:17] optimize them we can get within you know 100,000
[48:20] 100,000 fold these are all of the uh different
[48:23] fold these are all of the uh different uh species you went into at nartis
[48:26] uh species you went into at nartis different uh disease targets much of it
[48:29] different uh disease targets much of it didn't get published um there was uh we
[48:32] didn't get published um there was uh we worked with the US Army uh in 2003 Eola
[48:36] worked with the US Army uh in 2003 Eola had been weaponized by the US government
[48:38] had been weaponized by the US government and they wanted a vaccine and so we went
[48:41] and they wanted a vaccine and so we went into the aerosol uh challenge model uh
[48:44] into the aerosol uh challenge model uh we cloned in the glycoprotein uh from uh
[48:47] we cloned in the glycoprotein uh from uh Ebola it was very effective 100% uh
[48:50] Ebola it was very effective 100% uh protection uh didn't get to publish I
[48:53] protection uh didn't get to publish I showed everyone the data on malaria uh
[48:56] showed everyone the data on malaria uh from from Yale um uh
[49:03] yesterday just a list of all the
[49:05] yesterday just a list of all the Publications um but I often get asked
[49:09] Publications um but I often get asked well why did it take so long to publish
[49:11] well why did it take so long to publish well because in Industry you have to do
[49:13] well because in Industry you have to do a lot of work with the patent lawyers so
[49:15] a lot of work with the patent lawyers so I got a patent attorney to do a search
[49:18] I got a patent attorney to do a search Gil is the inventor uh over time
[49:21] Gil is the inventor uh over time applications a patent applications uh
[49:24] applications a patent applications uh grants are granted
[49:27] grants are granted by the the patent office and so what you
[49:30] by the the patent office and so what you can see 2008 the program began this is
[49:33] can see 2008 the program began this is some of my work on DNA vaccines you can
[49:35] some of my work on DNA vaccines you can see this huge Spike here uh and so I
[49:38] see this huge Spike here uh and so I have a massive patent portfolio I get no
[49:41] have a massive patent portfolio I get no money I signed off my rights uh and it's
[49:44] money I signed off my rights uh and it's it's nearly you reached its time of uh
[49:47] it's nearly you reached its time of uh patent life you know the cliff for these
[49:49] patent life you know the cliff for these are 2028 uh
[49:52] are 2028 uh 2030 but a lot of work from a lot of
[49:54] 2030 but a lot of work from a lot of scientists to file intellectual property
[49:57] scientists to file intellectual property and a lot of experiments to uh enable
[50:00] and a lot of experiments to uh enable these
[50:01] these claims as I said before 2015 the artist
[50:04] claims as I said before 2015 the artist decided to get out the vaccine game they
[50:06] decided to get out the vaccine game they wanted to focus on oncology so they did
[50:09] wanted to focus on oncology so they did some asset swaps with GSK uh I chose not
[50:13] some asset swaps with GSK uh I chose not to stay um uh they were moving everyone
[50:16] to stay um uh they were moving everyone from Boston down to Maryland madna were
[50:19] from Boston down to Maryland madna were hiring most of my team went to work for
[50:21] hiring most of my team went to work for madna uh I wanted to take some time out
[50:24] madna uh I wanted to take some time out and maybe do something different and let
[50:27] and maybe do something different and let the RNA Technologies mature a little bit
[50:29] the RNA Technologies mature a little bit and do some consultancy work uh and
[50:32] and do some consultancy work uh and that's what I did this is my list of
[50:36] that's what I did this is my list of what I thought needed to change when I
[50:38] what I thought needed to change when I left novatis so uh um they needed to be
[50:42] left novatis so uh um they needed to be better accessibility to lipid
[50:46] better accessibility to lipid nanoparticles um a scalable mixing
[50:49] nanoparticles um a scalable mixing technology was missing uh more potent
[50:52] technology was missing uh more potent vectors were needed uh reliable sources
[50:55] vectors were needed uh reliable sources of GMP enzyme times uh Downstream
[50:58] of GMP enzyme times uh Downstream equipment specifically designed for RNA
[51:01] equipment specifically designed for RNA and lmps cdmos that could manufacture
[51:05] and lmps cdmos that could manufacture RNA um to enable small companies to
[51:08] RNA um to enable small companies to innovate and uh there were a number of
[51:11] innovate and uh there were a number of analytical techniques that needed to
[51:13] analytical techniques that needed to come along covid solved most of these um
[51:17] come along covid solved most of these um and so small companies like replica we
[51:20] and so small companies like replica we don't have internal manufacturing we we
[51:23] don't have internal manufacturing we we Outsource it um and so we've gone from
[51:28] Outsource it um and so we've gone from designing novel vectors we began in 2020
[51:31] designing novel vectors we began in 2020 to having phase one clinical data um in
[51:34] to having phase one clinical data um in this year um so it's very rapid now the
[51:38] this year um so it's very rapid now the economics are much less building a clean
[51:41] economics are much less building a clean room for RNA hundred million now you can
[51:44] room for RNA hundred million now you can just Outsource it to a cdmo and they can
[51:47] just Outsource it to a cdmo and they can produce both the drug substance and the
[51:49] produce both the drug substance and the drug
[51:50] drug product so this is the detour uh for 5
[51:54] product so this is the detour uh for 5 years uh while I was Consulting
[51:57] years uh while I was Consulting um this is avidity biosciences this is
[51:59] um this is avidity biosciences this is the first publication down here it's a
[52:02] the first publication down here it's a Daran antibody conjugate highly potent
[52:05] Daran antibody conjugate highly potent sna we're in multiple um uh rare muscle
[52:10] sna we're in multiple um uh rare muscle diseases um I left as we were about to
[52:14] diseases um I left as we were about to uh um manufacture the phase one lot and
[52:18] uh um manufacture the phase one lot and file an indd uh for myotonic distrophy
[52:22] file an indd uh for myotonic distrophy now uh it's
[52:24] now uh it's 2025 uh they have they're in
[52:26] 2025 uh they have they're in registrational trials for three products
[52:28] registrational trials for three products so it's been pretty rapid uh the public
[52:30] so it's been pretty rapid uh the public is now uh the company is now public uh
[52:33] is now uh the company is now public uh worth over six billion dollars I was
[52:35] worth over six billion dollars I was part of the early crew the 25 people
[52:39] part of the early crew the 25 people that invented the technology we scaled
[52:41] that invented the technology we scaled it transitioned it into the clinic very
[52:44] it transitioned it into the clinic very proud of that work but it's a whole
[52:45] proud of that work but it's a whole another seminar but during that time I
[52:48] another seminar but during that time I was chairman of the board at Precision
[52:50] was chairman of the board at Precision assistants they became stiva I helped
[52:53] assistants they became stiva I helped them uh build out uh a lipid library
[52:57] them uh build out uh a lipid library that they wanted then to you know out
[52:59] that they wanted then to you know out license to the world and and create as
[53:02] license to the world and and create as research re agents which to me was kind
[53:04] research re agents which to me was kind of democratizing uh the the vaccine and
[53:07] of democratizing uh the the vaccine and gene therapy field uh they also worked
[53:10] gene therapy field uh they also worked on uh a modern uh microfluidic mixer
[53:14] on uh a modern uh microfluidic mixer this is the architecture they finally
[53:16] this is the architecture they finally came up with a toroidal mixer you can
[53:19] came up with a toroidal mixer you can change uh the diameter of of these uh
[53:22] change uh the diameter of of these uh inlets uh and change the flow rates that
[53:26] inlets uh and change the flow rates that can go through so at a small bench scale
[53:29] can go through so at a small bench scale at 1 millim minute you can make your
[53:30] at 1 millim minute you can make your preclinical material and then at GMP at
[53:33] preclinical material and then at GMP at 200 250 Ms a minute you can make 100
[53:36] 200 250 Ms a minute you can make 100 liters it's the same particles either
[53:40] liters it's the same particles either system so it's seamless way of scaling
[53:43] system so it's seamless way of scaling um it took many years of work and
[53:45] um it took many years of work and iterations with them uh to get it right
[53:47] iterations with them uh to get it right this is the system we use to produce our
[53:49] this is the system we use to produce our clinical material here's just a cryo em
[53:52] clinical material here's just a cryo em showing these solid dense particles and
[53:55] showing these solid dense particles and whatever scale you're at whichever of
[53:57] whatever scale you're at whichever of their devices um it it it produces the
[54:00] their devices um it it it produces the same material just allows us in a small
[54:03] same material just allows us in a small biotech to really move quickly out from
[54:05] biotech to really move quickly out from preclinical and get into the clinic uh
[54:08] preclinical and get into the clinic uh and show the technolog is working so
[54:11] and show the technolog is working so covid came in
[54:12] covid came in 2020 um I think everybody knows the
[54:15] 2020 um I think everybody knows the story but you know RNA vaccines are the
[54:18] story but you know RNA vaccines are the only ones that don't require cell
[54:20] only ones that don't require cell culture I've shown you speed it's fast
[54:23] culture I've shown you speed it's fast it's rapid you can clone from sequences
[54:26] it's rapid you can clone from sequences and move quickly but we got incredibly
[54:29] and move quickly but we got incredibly lucky uh because we knew a lot about the
[54:31] lucky uh because we knew a lot about the antigen and we had previous Corona virus
[54:35] antigen and we had previous Corona virus outbreaks Here's the the the main Front
[54:38] outbreaks Here's the the the main Front Runners uh Kovac had an unmodified RNA
[54:42] Runners uh Kovac had an unmodified RNA biion Tech uh fiser and madna with their
[54:46] biion Tech uh fiser and madna with their base modified and then over here um some
[54:50] base modified and then over here um some self-amplifying systems all of these are
[54:52] self-amplifying systems all of these are in lmps uh these guys all fell behind
[54:57] in lmps uh these guys all fell behind because of manufacturing issues
[54:58] because of manufacturing issues manufacturing longer RNA is very
[55:01] manufacturing longer RNA is very difficult compared to linear uh the
[55:03] difficult compared to linear uh the winners were in here with the base
[55:07] winners were in here with the base modified we' had outbreaks of SARS in 20
[55:12] modified we' had outbreaks of SARS in 20 2002 Ms this Middle Eastern variant in
[55:17] 2002 Ms this Middle Eastern variant in uh about 10 years later a lot of work
[55:19] uh about 10 years later a lot of work had gone in from the vaccine uh research
[55:22] had gone in from the vaccine uh research units particularly at the NIH on what's
[55:25] units particularly at the NIH on what's the key antigen they identified the
[55:27] the key antigen they identified the spike they were making prefusion
[55:29] spike they were making prefusion versions of it and here's just
[55:32] versions of it and here's just a an image the cryoem image of the that
[55:36] a an image the cryoem image of the that that Spike protein prefusion that came
[55:38] that Spike protein prefusion that came out from Barney grahames group so we got
[55:41] out from Barney grahames group so we got incredibly lucky and the RNA vaccines
[55:43] incredibly lucky and the RNA vaccines one if we didn't have that knowledge we
[55:47] one if we didn't have that knowledge we certainly would have have vaccines in
[55:49] certainly would have have vaccines in time and so antigen Discovery and trying
[55:52] time and so antigen Discovery and trying to survey what's out there is still very
[55:55] to survey what's out there is still very important just some timelines you know
[55:58] important just some timelines you know uh pandemic is starting to happen and
[56:01] uh pandemic is starting to happen and declared in December and we're starting
[56:03] declared in December and we're starting to have licensed vaccines less than a
[56:05] to have licensed vaccines less than a year later under emergency use
[56:08] year later under emergency use authorization normally vaccine approval
[56:10] authorization normally vaccine approval takes eight years it's certainly going
[56:12] takes eight years it's certainly going to slow down now we're probably looking
[56:14] to slow down now we're probably looking at four to five years in the US mRNA
[56:17] at four to five years in the US mRNA vaccines maybe never again we'll see
[56:19] vaccines maybe never again we'll see there's a lot of politics going
[56:22] there's a lot of politics going on this was the race in in the
[56:24] on this was the race in in the self-amplifying system
[56:26] self-amplifying system uh and we had a a a failure from
[56:30] uh and we had a a a failure from Imperial College London I I'm just going
[56:33] Imperial College London I I'm just going to talk a little bit about Urus the San
[56:35] to talk a little bit about Urus the San Diego company uh they they probably
[56:37] Diego company uh they they probably changed their vaccine four times during
[56:40] changed their vaccine four times during the clinical process optimizing the
[56:42] the clinical process optimizing the antigen optimizing manufacturing but
[56:44] antigen optimizing manufacturing but they finally got there um and in uh
[56:49] they finally got there um and in uh 2004 they got approval of the first
[56:52] 2004 they got approval of the first self-amplifying RNA vaccine in a lipid n
[56:55] self-amplifying RNA vaccine in a lipid n article as a CO
[56:56] article as a CO booster uh here's the paper they
[56:59] booster uh here's the paper they published um recently um they're
[57:04] published um recently um they're comparing it's a booster trial looking
[57:06] comparing it's a booster trial looking for non- inferiority they're comparing
[57:08] for non- inferiority they're comparing against the the biontech fiser vaccine
[57:11] against the the biontech fiser vaccine at 30 micrograms they're in at five so
[57:14] at 30 micrograms they're in at five so base modified versus
[57:16] base modified versus unmodified uh the safety profiles are
[57:19] unmodified uh the safety profiles are pretty similar they're seeing severe
[57:21] pretty similar they're seeing severe Adverse Events this may be under
[57:24] Adverse Events this may be under emergency use this is acceptable
[57:26] emergency use this is acceptable in my world uh we we need to be have a
[57:30] in my world uh we we need to be have a lot better safety
[57:33] lot better safety profile but did they did see better
[57:35] profile but did they did see better durability and so here we're looking at
[57:37] durability and so here we're looking at antibodies for a couple of different
[57:39] antibodies for a couple of different strains and looking at durability over
[57:41] strains and looking at durability over time out to day 181 uh so showing
[57:45] time out to day 181 uh so showing superiority in the durability and so
[57:48] superiority in the durability and so that's now you know a common thought
[57:50] that's now you know a common thought within the field is the self-amplifying
[57:52] within the field is the self-amplifying replicating systems will give you better
[57:55] replicating systems will give you better durability lower
[57:57] durability lower dust the other thing that came from the
[58:00] dust the other thing that came from the pandemic we got started getting modeling
[58:02] pandemic we got started getting modeling systems uh this one came from Imperial
[58:04] systems uh this one came from Imperial College London uh and their modeling
[58:07] College London uh and their modeling system is fairly crude but the here are
[58:09] system is fairly crude but the here are the different RNA Technologies here are
[58:12] the different RNA Technologies here are the proposed doses uh whether they're
[58:14] the proposed doses uh whether they're using modified bases or not um and the
[58:18] using modified bases or not um and the common size of a bioreactor you'd expect
[58:20] common size of a bioreactor you'd expect to use if you have one Factory how long
[58:23] to use if you have one Factory how long does it take to vaccinate the world and
[58:25] does it take to vaccinate the world and no surprises if you can get down to 1.1
[58:29] no surprises if you can get down to 1.1 microgram it's going to be a lot
[58:31] microgram it's going to be a lot easier so replicate we found it in 2020
[58:36] easier so replicate we found it in 2020 I took a year sabatical and worked with
[58:38] I took a year sabatical and worked with the group in Vancouver I never went
[58:40] the group in Vancouver I never went there uh because of covid but
[58:42] there uh because of covid but coordinated and got the lipid systems
[58:45] coordinated and got the lipid systems ready the the the microfluid technology
[58:48] ready the the the microfluid technology ready there um and came on board in in
[58:51] ready there um and came on board in in 2021 we're still 19 employees uh we
[58:55] 2021 we're still 19 employees uh we focused on develop these second
[58:57] focused on develop these second generation vectors that are more potent
[59:00] generation vectors that are more potent than the first generation
[59:01] than the first generation self-amplifying uh going to show you
[59:03] self-amplifying uh going to show you some phase one
[59:05] some phase one data we the sequencing data for uh the
[59:09] data we the sequencing data for uh the alpha virus species got dramatically
[59:11] alpha virus species got dramatically better it's one of the reasons we
[59:13] better it's one of the reasons we founded the company we felt there was an
[59:15] founded the company we felt there was an opportunity to pull out novel uh Vector
[59:19] opportunity to pull out novel uh Vector designs that might be more potent
[59:21] designs that might be more potent opportunity to file IP raise money it's
[59:24] opportunity to file IP raise money it's the biotech mantra in the uh the
[59:28] the biotech mantra in the uh the US most if not all those first
[59:31] US most if not all those first generation use a common Vector uh from
[59:34] generation use a common Vector uh from Venezuelan Ean and sephtis uh tc83 it's
[59:38] Venezuelan Ean and sephtis uh tc83 it's derived from the live attenuated vaccine
[59:41] derived from the live attenuated vaccine developed by the US Army in the 90s they
[59:44] developed by the US Army in the 90s they published the paper with the sequence
[59:46] published the paper with the sequence before they filed the patent and
[59:48] before they filed the patent and therefore that is freely available to
[59:51] therefore that is freely available to everyone to use um but there are over 45
[59:54] everyone to use um but there are over 45 species of alpha virus sequencing datas
[59:57] species of alpha virus sequencing datas better so we start fishing out novel
[01:00:00] better so we start fishing out novel variants getting them and launching them
[01:00:02] variants getting them and launching them in uh cell culture filing patterns and
[01:00:05] in uh cell culture filing patterns and we found a way of optimizing the three
[01:00:08] we found a way of optimizing the three and five Prime ends to make them a
[01:00:10] and five Prime ends to make them a little more potent and easier to
[01:00:12] little more potent and easier to manufacture
[01:00:14] manufacture so here's some data this is an
[01:00:17] so here's some data this is an independent study that came from uh uh
[01:00:20] independent study that came from uh uh Anna blakey's lab she's a co-author on
[01:00:22] Anna blakey's lab she's a co-author on this paper uh so we she had uh the tc83
[01:00:27] this paper uh so we she had uh the tc83 vector we're encoding luciferase we
[01:00:29] vector we're encoding luciferase we Center over an optimized V Vector so
[01:00:32] Center over an optimized V Vector so we're comparing species to species the
[01:00:35] we're comparing species to species the in the same LMP they're dosed at the
[01:00:37] in the same LMP they're dosed at the same dose and we're measuring total flux
[01:00:40] same dose and we're measuring total flux bioluminescence over time and what you
[01:00:42] bioluminescence over time and what you can see is uh higher levels of
[01:00:46] can see is uh higher levels of bioluminescence and better durability
[01:00:49] bioluminescence and better durability typically we see uh out to 30 40 Days
[01:00:52] typically we see uh out to 30 40 Days for these self-replicating systems
[01:00:56] for these self-replicating systems when we went into a mouse model uh with
[01:00:59] when we went into a mouse model uh with flu remember in the previous work at
[01:01:02] flu remember in the previous work at nartis we're dosing a one or 0.1
[01:01:04] nartis we're dosing a one or 0.1 microgram here we're dosing uh at 10
[01:01:09] microgram here we're dosing uh at 10 nanog down to one picogram uh We've
[01:01:13] nanog down to one picogram uh We've either got an H5 variant from a Vietnam
[01:01:17] either got an H5 variant from a Vietnam or the H1 a California strain uh we're
[01:01:20] or the H1 a California strain uh we're doing a prime or a boost we're measuring
[01:01:23] doing a prime or a boost we're measuring hemog gluten inhibition Titus and and so
[01:01:26] hemog gluten inhibition Titus and and so you can see the M of priming even at
[01:01:28] you can see the M of priming even at these low doses little better for the H5
[01:01:31] these low doses little better for the H5 antigen and then we boost for the H5
[01:01:35] antigen and then we boost for the H5 we're not seeing a dose response until
[01:01:37] we're not seeing a dose response until we're down at one peam so we've pushed
[01:01:39] we're down at one peam so we've pushed the technology we've got a delivery
[01:01:42] the technology we've got a delivery system in licensed from Precision Nano
[01:01:45] system in licensed from Precision Nano systems we've optimized the vector and
[01:01:47] systems we've optimized the vector and we've gone from one or 0.1 microgram now
[01:01:51] we've gone from one or 0.1 microgram now down to uh a picogram
[01:01:56] we chose to use a cdmo network to not
[01:02:00] we chose to use a cdmo network to not build the infrastructure ourselves it
[01:02:02] build the infrastructure ourselves it was quite a collaboration uh they
[01:02:04] was quite a collaboration uh they weren't ready to go so at curia we spent
[01:02:07] weren't ready to go so at curia we spent a couple of years teaching them how to
[01:02:08] a couple of years teaching them how to make long RNA and the way you do that is
[01:02:11] make long RNA and the way you do that is you have assays that tell you if it's
[01:02:13] you have assays that tell you if it's working we have a cell-based potency
[01:02:15] working we have a cell-based potency assay we have a capillary gel electris
[01:02:18] assay we have a capillary gel electris system to measure integrity and we
[01:02:20] system to measure integrity and we probed the process all the way through
[01:02:23] probed the process all the way through to figure out where we were seeing
[01:02:24] to figure out where we were seeing degradation longer rnas will hydrolize
[01:02:27] degradation longer rnas will hydrolize faster and time at temperature and pH
[01:02:30] faster and time at temperature and pH are important so we optimize the
[01:02:32] are important so we optimize the conditions to keep potency and we did
[01:02:34] conditions to keep potency and we did the same with Precision Nanos systems
[01:02:36] the same with Precision Nanos systems who are ntiva but we en licensed the
[01:02:39] who are ntiva but we en licensed the lipid from them so we didn't have to
[01:02:42] lipid from them so we didn't have to build that capacity ourselves so we're
[01:02:43] build that capacity ourselves so we're we're saving time by incensing the lipid
[01:02:47] we're saving time by incensing the lipid Outsourcing the manufacturing but using
[01:02:49] Outsourcing the manufacturing but using our skills to teach them what to do um
[01:02:52] our skills to teach them what to do um and the the lipid systems we used went
[01:02:56] and the the lipid systems we used went through safety talkx testing genotox
[01:03:00] through safety talkx testing genotox testing under the Canadian government at
[01:03:02] testing under the Canadian government at the beginning of
[01:03:03] the beginning of covid so this is the clean this is the
[01:03:07] covid so this is the clean this is the the RBI 4000 our rabies vaccine it's our
[01:03:11] the RBI 4000 our rabies vaccine it's our uh V Vector with the glycoprotein from
[01:03:14] uh V Vector with the glycoprotein from rabies in and we're comparing it to RAB
[01:03:17] rabies in and we're comparing it to RAB the whole inactivated virus so the the
[01:03:20] the whole inactivated virus so the the full protein
[01:03:21] full protein repertoire um why rabies This was meant
[01:03:25] repertoire um why rabies This was meant as a proof of concept for the platform
[01:03:27] as a proof of concept for the platform so in the US nobody has rabies so it's a
[01:03:31] so in the US nobody has rabies so it's a clean patient population can we Prime
[01:03:34] clean patient population can we Prime can we boost that response in covid
[01:03:36] can we boost that response in covid everyone is positive it's a very
[01:03:39] everyone is positive it's a very complicated uh uh immunological
[01:03:42] complicated uh uh immunological interpretation of what's going on uh so
[01:03:44] interpretation of what's going on uh so we chose not to do flu we chose not to
[01:03:46] we chose not to do flu we chose not to do covid um there's also a World Health
[01:03:49] do covid um there's also a World Health Organization established C of protection
[01:03:52] Organization established C of protection in a new assay if you're above 05
[01:03:54] in a new assay if you're above 05 International units per Mill you're
[01:03:56] International units per Mill you're considered protective there's a
[01:03:58] considered protective there's a commercial product to Benchmark
[01:04:01] commercial product to Benchmark against so here's the trial design 0.1
[01:04:05] against so here's the trial design 0.1 one and 10 micrograms two doses eight
[01:04:08] one and 10 micrograms two doses eight weeks
[01:04:09] weeks apart cohort 3 is the anomaly it's a
[01:04:12] apart cohort 3 is the anomaly it's a single dose um and then rabbert is
[01:04:17] single dose um and then rabbert is cohort 5 the standard CDC guidelines now
[01:04:20] cohort 5 the standard CDC guidelines now are two doses for a prophylactic vaccine
[01:04:23] are two doses for a prophylactic vaccine uh one week about about so we're dosing
[01:04:24] uh one week about about so we're dosing eight weeks
[01:04:26] eight weeks one week for RAB primary endpoint is
[01:04:30] one week for RAB primary endpoint is safety reactogenicity determining that
[01:04:33] safety reactogenicity determining that uh dose for phase two secondary endpoint
[01:04:35] uh dose for phase two secondary endpoint is uh the new assay and exploratory te-
[01:04:39] is uh the new assay and exploratory te- cells healthy people 18 to 45 serer
[01:04:43] cells healthy people 18 to 45 serer negative for viral infection normal
[01:04:45] negative for viral infection normal blood uh no prior rabies vaccination or
[01:04:48] blood uh no prior rabies vaccination or exposure there were three people that
[01:04:51] exposure there were three people that were positive for rabies they were
[01:04:53] were positive for rabies they were pulled from The Trial I'll show you the
[01:04:54] pulled from The Trial I'll show you the data at the end it's very interesting uh
[01:04:57] data at the end it's very interesting uh it's published so you can go read about
[01:04:59] it's published so you can go read about it but this is the the overall summary
[01:05:03] it but this is the the overall summary of the data out to day
[01:05:06] of the data out to day 84 so in Gray this means in that group
[01:05:10] 84 so in Gray this means in that group they've got detectable neutralizing
[01:05:11] they've got detectable neutralizing antibodies above Baseline blue means
[01:05:14] antibodies above Baseline blue means they've gone to or above serero
[01:05:17] they've gone to or above serero conversion uh for that surrogate of
[01:05:19] conversion uh for that surrogate of protection so even at 0.1 microgram
[01:05:22] protection so even at 0.1 microgram we're priming the majority we've got
[01:05:24] we're priming the majority we've got measurable responses some of them are
[01:05:26] measurable responses some of them are protected we boost at 0.1 the majority
[01:05:29] protected we boost at 0.1 the majority are protected nearly 100% have Ser
[01:05:33] are protected nearly 100% have Ser converted if this was covid this would
[01:05:36] converted if this was covid this would probably be viable 0.1 microgram we're
[01:05:39] probably be viable 0.1 microgram we're at 50 million human doses per liter in a
[01:05:42] at 50 million human doses per liter in a few hours so this changes the world of
[01:05:46] few hours so this changes the world of pandemic
[01:05:47] pandemic response one microgram 100% two doses uh
[01:05:51] response one microgram 100% two doses uh nearly all of them are protected even a
[01:05:54] nearly all of them are protected even a single dose
[01:05:56] single dose uh conventional linear RNA with base
[01:05:58] uh conventional linear RNA with base modifications if you look they're barely
[01:06:01] modifications if you look they're barely priming in that new response here with
[01:06:04] priming in that new response here with the single dose we've nearly we've Ser
[01:06:07] the single dose we've nearly we've Ser converted everybody they've got use but
[01:06:09] converted everybody they've got use but nearly all of them are protected two
[01:06:13] nearly all of them are protected two doses 100% protection and we can match
[01:06:16] doses 100% protection and we can match rabber it so here are the kinetics out
[01:06:19] rabber it so here are the kinetics out to day 84 and you can see no rabbert
[01:06:23] to day 84 and you can see no rabbert comes up Prime in a boost and then the
[01:06:25] comes up Prime in a boost and then the Titus start to Decay our RNA Titus come
[01:06:28] Titus start to Decay our RNA Titus come up and then we boost and they all come
[01:06:31] up and then we boost and they all come up nicely we're monitoring out we're
[01:06:34] up nicely we're monitoring out we're just rounding off the data sets to day
[01:06:36] just rounding off the data sets to day 240 it looks like two doses at 10
[01:06:39] 240 it looks like two doses at 10 micrograms are superior in durability to
[01:06:43] micrograms are superior in durability to RAB if we look at post Prime what
[01:06:47] RAB if we look at post Prime what happens with the RNA at day 28 where we
[01:06:49] happens with the RNA at day 28 where we see the maximum titis we see a nice dose
[01:06:52] see the maximum titis we see a nice dose response in in the new assay here's the
[01:06:55] response in in the new assay here's the correlate of protection there's that one
[01:06:57] correlate of protection there's that one that didn't zero
[01:06:59] that didn't zero convert we're kind of come on to the
[01:07:01] convert we're kind of come on to the safety data but what we see from the
[01:07:03] safety data but what we see from the safety data is no dose limiting tox no
[01:07:06] safety data is no dose limiting tox no severe Adverse Events so we could dose
[01:07:08] severe Adverse Events so we could dose higher so in phase two we're going to
[01:07:12] higher so in phase two we're going to continue this dose escalation can we
[01:07:15] continue this dose escalation can we dose 15 or 20 micrograms is it tolerated
[01:07:18] dose 15 or 20 micrograms is it tolerated and can we push uh a single dose to the
[01:07:21] and can we push uh a single dose to the level of rabber over here so when we're
[01:07:24] level of rabber over here so when we're boosting this is RT at day uh 18 um and
[01:07:29] boosting this is RT at day uh 18 um and here's our two doses of 10 microgram uh
[01:07:33] here's our two doses of 10 microgram uh at day 56 the maximum titles we're
[01:07:35] at day 56 the maximum titles we're seeing statistically these are
[01:07:37] seeing statistically these are equivalent obviously you know these guys
[01:07:40] equivalent obviously you know these guys here Ser convert at a different time
[01:07:43] here Ser convert at a different time point so we're starting to look very
[01:07:46] point so we're starting to look very competitive with a commercial vaccine
[01:07:48] competitive with a commercial vaccine we've got the opportunity to do single
[01:07:50] we've got the opportunity to do single dosing we're in a place where other RNA
[01:07:53] dosing we're in a place where other RNA Technologies are not
[01:07:56] Technologies are not safety wise uh this is uh the local
[01:08:00] safety wise uh this is uh the local reactions um either uh first dose or
[01:08:04] reactions um either uh first dose or second dose uh the thing to note is
[01:08:06] second dose uh the thing to note is there are no grade three reactions there
[01:08:08] there are no grade three reactions there are certainly no severe reactions or any
[01:08:11] are certainly no severe reactions or any dose limiting tox uh it's a small
[01:08:14] dose limiting tox uh it's a small patient participant group it's 18 people
[01:08:17] patient participant group it's 18 people uh if we start maybe the second dose
[01:08:20] uh if we start maybe the second dose looks less reactive than the first uh
[01:08:23] looks less reactive than the first uh this is certainly not what we see with
[01:08:25] this is certainly not what we see with other platforms usually reactor dasty
[01:08:27] other platforms usually reactor dasty goes up uh but they're W they're very
[01:08:30] goes up uh but they're W they're very mild transient and people recover within
[01:08:33] mild transient and people recover within a few
[01:08:34] a few days this is uh systemic reactions we
[01:08:37] days this is uh systemic reactions we saw one uh elevated temperature in the
[01:08:39] saw one uh elevated temperature in the middle dose cohort that was reported
[01:08:42] middle dose cohort that was reported from home 103 temperature for 4 hours
[01:08:45] from home 103 temperature for 4 hours there were no other symptoms the
[01:08:47] there were no other symptoms the participant got the booster reported no
[01:08:50] participant got the booster reported no adverse events at all the physician
[01:08:52] adverse events at all the physician thinks they misread the thermometer it's
[01:08:54] thinks they misread the thermometer it's reported we don't think it's real uh but
[01:08:57] reported we don't think it's real uh but overall uh grade one and grade two and
[01:09:00] overall uh grade one and grade two and again those drop on the second dose so
[01:09:03] again those drop on the second dose so from a vaccine perspective in the MRNA
[01:09:05] from a vaccine perspective in the MRNA World we've got potency we've got great
[01:09:08] World we've got potency we've got great uh tolerability data uh and we're very
[01:09:13] uh tolerability data uh and we're very competitive three people were positive
[01:09:15] competitive three people were positive at Baseline which was unexpected for us
[01:09:19] at Baseline which was unexpected for us uh we they got the the first shot um and
[01:09:22] uh we they got the the first shot um and but what you can see is we got a
[01:09:24] but what you can see is we got a dramatic uh increase in titer 13 fold
[01:09:28] dramatic uh increase in titer 13 fold there was one in each dose cohort and
[01:09:30] there was one in each dose cohort and the titter uh remain very high haven't
[01:09:34] the titter uh remain very high haven't come down and we continue to monitor
[01:09:36] come down and we continue to monitor them uh here's the the surrogate level
[01:09:39] them uh here's the the surrogate level of
[01:09:44] protection
[01:09:49] no we think so so I know you guys have
[01:09:52] no we think so so I know you guys have been exploring Prime boost strategies
[01:09:55] been exploring Prime boost strategies [Laughter]
[01:09:58] [Laughter] you know and certainly it's a way of do
[01:10:00] you know and certainly it's a way of do of dosing you know uh vaccines you Prime
[01:10:04] of dosing you know uh vaccines you Prime with one antigen boost with another um
[01:10:07] with one antigen boost with another um or you can mix them together but we were
[01:10:09] or you can mix them together but we were surprised by the data we don't know what
[01:10:11] surprised by the data we don't know what why they were Sero positive our
[01:10:13] why they were Sero positive our assumption is they were in the military
[01:10:15] assumption is they were in the military or a vet tech assistant and they've been
[01:10:17] or a vet tech assistant and they've been given a conventional vaccine but we we
[01:10:20] given a conventional vaccine but we we love the data no dose response 01110
[01:10:23] love the data no dose response 01110 micrograms pretty similar and they keep
[01:10:26] micrograms pretty similar and they keep going out over
[01:10:28] going out over time uh so overall uh clean safety
[01:10:32] time uh so overall uh clean safety profile we we're in very good shape uh
[01:10:36] profile we we're in very good shape uh uh to do continue the program we're not
[01:10:39] uh to do continue the program we're not continuing the program we're in active
[01:10:41] continuing the program we're in active discussions with people in in South
[01:10:43] discussions with people in in South America uh potentially somebody in
[01:10:45] America uh potentially somebody in Brazil uh and hopefully we can get the
[01:10:48] Brazil uh and hopefully we can get the program uh through Phase 2 three and
[01:10:51] program uh through Phase 2 three and lure both as a prophylactic and a
[01:10:53] lure both as a prophylactic and a postexposure uh strategy um for uh South
[01:10:58] postexposure uh strategy um for uh South America uh but we're definitely seeing
[01:11:00] America uh but we're definitely seeing improved
[01:11:01] improved bioactivity potential of a single dose
[01:11:04] bioactivity potential of a single dose everything that we could have helped for
[01:11:05] everything that we could have helped for hoped for for improvements in the
[01:11:09] hoped for for improvements in the technology so overall how do we compare
[01:11:11] technology so overall how do we compare to the other vaccine most of the data is
[01:11:14] to the other vaccine most of the data is in covid we're in in in rabies so the
[01:11:18] in covid we're in in in rabies so the easiest way to compare is therapeutic
[01:11:21] easiest way to compare is therapeutic index highest clinical dose reported
[01:11:23] index highest clinical dose reported with favorable safety profile divided by
[01:11:26] with favorable safety profile divided by the lowest clinical dose showing 50%
[01:11:29] the lowest clinical dose showing 50% protection and greater than 100 because
[01:11:32] protection and greater than 100 because we didn't see any talks and everyone
[01:11:35] we didn't see any talks and everyone else whether they're self-amplifying
[01:11:37] else whether they're self-amplifying with modifications without they're all
[01:11:40] with modifications without they're all around 3x so they're all dosing as high
[01:11:43] around 3x so they're all dosing as high as they can till they hit tolerability
[01:11:45] as they can till they hit tolerability issues and that's their dose that's not
[01:11:47] issues and that's their dose that's not what where we're at we've we think we're
[01:11:49] what where we're at we've we think we're in a competitive
[01:11:51] in a competitive space um this is our
[01:11:53] space um this is our pipeline so we're hoping to move rabies
[01:11:57] pipeline so we're hoping to move rabies forward uh our lead program is in
[01:11:59] forward uh our lead program is in Epstein bar virus uh we're going to go
[01:12:02] Epstein bar virus uh we're going to go in and TR treat infectious mon nucleosis
[01:12:05] in and TR treat infectious mon nucleosis in the Adolescent po population this has
[01:12:08] in the Adolescent po population this has five antigens so this is out pushing out
[01:12:11] five antigens so this is out pushing out to 16 KB this is where we think there's
[01:12:13] to 16 KB this is where we think there's a unique opportunity with the technology
[01:12:16] a unique opportunity with the technology one vector five antigens or six antigens
[01:12:20] one vector five antigens or six antigens uh so and hopefully we can optimize the
[01:12:24] uh so and hopefully we can optimize the cassette and the sequence and how we
[01:12:26] cassette and the sequence and how we express the antigens to get equal
[01:12:29] express the antigens to get equal potency from all of the
[01:12:31] potency from all of the antigens we have some undisclosed
[01:12:33] antigens we have some undisclosed targets we have some prary data in uh
[01:12:36] targets we have some prary data in uh infl inflammatory disease we have a a
[01:12:39] infl inflammatory disease we have a a breast cancer vaccine parked up ready
[01:12:41] breast cancer vaccine parked up ready waiting for funding where we're going
[01:12:43] waiting for funding where we're going after uh mutations known mutations that
[01:12:46] after uh mutations known mutations that occur in estrogen positive uh uh breast
[01:12:50] occur in estrogen positive uh uh breast cancer when they're on hormonal
[01:12:52] cancer when they're on hormonal therapies uh and so if the cells mutate
[01:12:55] therapies uh and so if the cells mutate T sales will take them
[01:12:58] out with that just like to say thank you
[01:13:01] out with that just like to say thank you to the 100,000 probably people I've
[01:13:03] to the 100,000 probably people I've worked with this is the team at replica
[01:13:06] worked with this is the team at replica uh thanks to all the study volunteers
[01:13:08] uh thanks to all the study volunteers you know uh the site Personnel Anna
[01:13:11] you know uh the site Personnel Anna Blakey and Credo are up at UBC hopefully
[01:13:14] Blakey and Credo are up at UBC hopefully I can get them talking to you uh you
[01:13:16] I can get them talking to you uh you know she's a she's a self-amplifying RNA
[01:13:19] know she's a she's a self-amplifying RNA expert knows about delivery uh she'd be
[01:13:22] expert knows about delivery uh she'd be a great place to send interns to to do
[01:13:25] a great place to send interns to to do training on um thanks to the clinical
[01:13:28] training on um thanks to the clinical people clinical sites uh and and our
[01:13:31] people clinical sites uh and and our cdmos who who made the
[01:13:33] cdmos who who made the material and that's me that's my wife
[01:13:38] material and that's me that's my wife somebody's taking a picture of me taking
[01:13:40] somebody's taking a picture of me taking a picture of the
[01:13:46] turtle next time you should go
[01:13:50] turtle next time you should go to
[01:13:53] Great okay okay uh thank you Andy uh do
[01:13:57] Great okay okay uh thank you Andy uh do we have
[01:14:00] questions we we've been discussing for
[01:14:03] questions we we've been discussing for two days now but I still have few
[01:14:06] two days now but I still have few questions regarding the optimization of
[01:14:09] questions regarding the optimization of the vector yes because that's a big
[01:14:12] the vector yes because that's a big issue if you want to have like four five
[01:14:15] issue if you want to have like four five six antigens do you really need or all
[01:14:19] six antigens do you really need or all four nonstructural proteins because only
[01:14:22] four nonstructural proteins because only one is the RNA replicates isn't it
[01:14:25] one is the RNA replicates isn't it why you need all four NS protein the the
[01:14:30] why you need all four NS protein the the biology is quite complicated uh you've
[01:14:33] biology is quite complicated uh you've got to cap You' got to put a polyat tail
[01:14:36] got to cap You' got to put a polyat tail on you've got to get it to cleave off
[01:14:38] on you've got to get it to cleave off the subgenomic promoter but those
[01:14:41] the subgenomic promoter but those proteins also go into the host cell
[01:14:44] proteins also go into the host cell genome yeah and they co-opt and you
[01:14:47] genome yeah and they co-opt and you suppress antiviral responses you skew
[01:14:50] suppress antiviral responses you skew pro protein expression towards the the
[01:14:53] pro protein expression towards the the the viral genome needs so they do a lot
[01:14:56] the viral genome needs so they do a lot and but we so we now have 17 sets of
[01:15:00] and but we so we now have 17 sets of non-structural proteins that do
[01:15:02] non-structural proteins that do different things so if if we want a t-
[01:15:05] different things so if if we want a t- cell vaccine uh we we screen and it's
[01:15:10] cell vaccine uh we we screen and it's usually not V it's something
[01:15:12] usually not V it's something else yeah I was wondering if you can
[01:15:15] else yeah I was wondering if you can make some sort of deletions in this NS
[01:15:18] make some sort of deletions in this NS protein to make the whole thing smaller
[01:15:21] protein to make the whole thing smaller and it not sometimes one deletion kills
[01:15:25] and it not sometimes one deletion kills SK the whole thing and and so people
[01:15:27] SK the whole thing and and so people have been trying for years to to do that
[01:15:30] have been trying for years to to do that and we we've like we we've just
[01:15:34] and we we've like we we've just said that's the way it is we're going to
[01:15:36] said that's the way it is we're going to go with a a library of different vectors
[01:15:39] go with a a library of different vectors with different properties and explore
[01:15:40] with different properties and explore that rather than trying to do point
[01:15:42] that rather than trying to do point mutations
[01:15:44] mutations or okay and second quick question how
[01:15:47] or okay and second quick question how much uh plasma DNA you need to start
[01:15:50] much uh plasma DNA you need to start your Biore reator like for one liter how
[01:15:53] your Biore reator like for one liter how much DNA you need to put on
[01:15:55] much DNA you need to put on it's like a 1 to 300
[01:16:06] conversion yeah something like that I'm
[01:16:09] conversion yeah something like that I'm I don't have that number no it's not a
[01:16:12] I don't have that number no it's not a huge
[01:16:17] amount yeah yeah in a liter we can make
[01:16:20] amount yeah yeah in a liter we can make five grams of of of RNA
[01:16:25] yeah so it's about you divide by two or
[01:16:29] yeah so it's about you divide by two or 300 and that's the amount of plasmid you
[01:16:32] 300 and that's the amount of plasmid you need but you know we're still we
[01:16:35] need but you know we're still we we we're we're always optimizing the
[01:16:37] we we're we're always optimizing the plasmids and so now we're looking at
[01:16:39] plasmids and so now we're looking at synthetic plasmids where they're a lot
[01:16:42] synthetic plasmids where they're a lot simpler that you haven't got any of the
[01:16:44] simpler that you haven't got any of the spinach you need to grow them in eoli uh
[01:16:47] spinach you need to grow them in eoli uh maybe we'll need less of them uh they're
[01:16:50] maybe we'll need less of them uh they're not quite ready for 9 16 KB yet but it's
[01:16:54] not quite ready for 9 16 KB yet but it's coming
[01:16:55] coming and and then you we can it's a lot
[01:16:59] and and then you we can it's a lot faster for us it's 3 four months to make
[01:17:03] faster for us it's 3 four months to make a GMP compliant batch of plasmid
[01:17:06] a GMP compliant batch of plasmid synthetically you could do it in
[01:17:08] synthetically you could do it in probably three
[01:17:12] weeks thank you very much for the
[01:17:14] weeks thank you very much for the presentation Dr Gil uh I'm a pharmacist
[01:17:17] presentation Dr Gil uh I'm a pharmacist not an immunologist so I'm aist yeah I'm
[01:17:20] not an immunologist so I'm aist yeah I'm going to ask you something about the uh
[01:17:23] going to ask you something about the uh the RNA production uh how much more
[01:17:26] the RNA production uh how much more expensive or more complicated is to make
[01:17:29] expensive or more complicated is to make this self-replicating RNA compared to
[01:17:32] this self-replicating RNA compared to the m it's the same recipe okay you it
[01:17:35] the m it's the same recipe okay you it you just got to play around with time
[01:17:38] you just got to play around with time and temperature and the sequence of
[01:17:40] and temperature and the sequence of addition and it's it's it's very similar
[01:17:45] addition and it's it's it's very similar um and you know we we're kind of do I'm
[01:17:48] um and you know we we're kind of do I'm back and forth now with some commercial
[01:17:50] back and forth now with some commercial enzyme producers and they think you know
[01:17:53] enzyme producers and they think you know if a dose
[01:17:55] if a dose with the self let's say it was 10
[01:17:56] with the self let's say it was 10 micrograms they think they can get below
[01:17:59] micrograms they think they can get below a dollar for so we we think we can the
[01:18:02] a dollar for so we we think we can the cost of goods is going to be very low
[01:18:04] cost of goods is going to be very low because the doses are lower and and our
[01:18:06] because the doses are lower and and our aim is to try and get to less than a
[01:18:08] aim is to try and get to less than a dollar so so that you know it it then
[01:18:12] dollar so so that you know it it then it's a it's cost effective in in South
[01:18:16] it's a it's cost effective in in South America and Beyond because you can't we
[01:18:18] America and Beyond because you can't we can't be charging $100 for a vaccine
[01:18:21] can't be charging $100 for a vaccine here yes $10 would kill everyone
[01:18:24] here yes $10 would kill everyone probably it needs to be a couple of
[01:18:27] probably it needs to be a couple of dollar okay and the other quick question
[01:18:30] dollar okay and the other quick question is uh in some point in our presentation
[01:18:32] is uh in some point in our presentation you showed when you were uh doing the
[01:18:34] you showed when you were uh doing the Consulting work uh you have uh you
[01:18:37] Consulting work uh you have uh you listed the challenges uh for developing
[01:18:41] listed the challenges uh for developing the vaccines by that time but uh now
[01:18:44] the vaccines by that time but uh now what's what would you consider the most
[01:18:47] what's what would you consider the most uh the biggest challenge that we have to
[01:18:49] uh the biggest challenge that we have to face to expand the pipelines of uh Mr
[01:18:53] face to expand the pipelines of uh Mr vaccines or
[01:18:55] vaccines or antigens good antigens antigens yeah so
[01:18:59] antigens good antigens antigens yeah so antigen Discovery so CMV is a huge unmet
[01:19:03] antigen Discovery so CMV is a huge unmet need in the developing world I think it
[01:19:05] need in the developing world I think it it's 96 proteins in the genome which one
[01:19:10] it's 96 proteins in the genome which one which one do you need for the perfect
[01:19:12] which one do you need for the perfect vaccine so it's always antigen design
[01:19:16] vaccine so it's always antigen design and finding the right antigen um to to
[01:19:19] and finding the right antigen um to to go so I think we have a great platform
[01:19:22] go so I think we have a great platform um and it's now for us it's finding the
[01:19:26] um and it's now for us it's finding the right antigens to par we're not going to
[01:19:28] right antigens to par we're not going to be able to tackle every infectious
[01:19:30] be able to tackle every infectious disease but there are many we can tackle
[01:19:33] disease but there are many we can tackle quite easily you know for instance
[01:19:35] quite easily you know for instance seasonal flu putting in four
[01:19:38] seasonal flu putting in four Haas not a problem but if those ha
[01:19:41] Haas not a problem but if those ha constructs are not designed well the
[01:19:43] constructs are not designed well the vaccine won't
[01:19:46] work uh thank you for your presentation
[01:19:49] work uh thank you for your presentation it was really good and I have two
[01:19:51] it was really good and I have two questions uh the first one is uh for how
[01:19:55] questions uh the first one is uh for how long does this uh self replicating mRNA
[01:19:59] long does this uh self replicating mRNA they they belong they still there in the
[01:20:01] they they belong they still there in the cell and uh could you measure the the
[01:20:05] cell and uh could you measure the the the amount uh over the
[01:20:08] the amount uh over the time
[01:20:09] time so in the mouse with luciferase it keeps
[01:20:13] so in the mouse with luciferase it keeps we see bioluminesence for um 30 40 days
[01:20:18] we see bioluminesence for um 30 40 days um and the half life of Lucifer is ours
[01:20:22] um and the half life of Lucifer is ours so but when we put an pen in it's much
[01:20:26] so but when we put an pen in it's much shorter so there's a published
[01:20:28] shorter so there's a published experiment in 2017 from the team at
[01:20:30] experiment in 2017 from the team at nartis where we coex we did luciferase
[01:20:34] nartis where we coex we did luciferase alone we did luciferase with uh a
[01:20:37] alone we did luciferase with uh a bicistronic with rvf and the
[01:20:40] bicistronic with rvf and the bioluminescence truncated down so when
[01:20:42] bioluminescence truncated down so when you put a glycoprotein on the surface
[01:20:45] you put a glycoprotein on the surface the t- cells are going to see it they're
[01:20:46] the t- cells are going to see it they're going to come in t- cell positive for
[01:20:48] going to come in t- cell positive for the antigen around day seven and that's
[01:20:51] the antigen around day seven and that's when you start to see uh clearance of
[01:20:54] when you start to see uh clearance of those
[01:20:55] those transfected cells in cell
[01:20:58] transfected cells in cell culture 3 or 4 days and and they go into
[01:21:01] culture 3 or 4 days and and they go into apoptosis because it spills out into the
[01:21:04] apoptosis because it spills out into the cytoplasm and it goes into apoptosis
[01:21:08] cytoplasm and it goes into apoptosis yeah so the second question it it was
[01:21:11] yeah so the second question it it was dependent of this this answer so uh do
[01:21:15] dependent of this this answer so uh do you think that you can extend the the
[01:21:19] you think that you can extend the the the the duration of this yeah that's one
[01:21:22] the the duration of this yeah that's one of the reasons we went in to look at the
[01:21:24] of the reasons we went in to look at the different species they all have
[01:21:26] different species they all have different patterns of the way they
[01:21:29] different patterns of the way they replicate and and some of them replicate
[01:21:32] replicate and and some of them replicate at a lower level but they go on for
[01:21:34] at a lower level but they go on for months yeah and uh doing that do you
[01:21:38] months yeah and uh doing that do you think that you can use this this kind of
[01:21:41] think that you can use this this kind of technology to deliver proteins to cells
[01:21:44] technology to deliver proteins to cells that's so so behind behind the vaccine
[01:21:47] that's so so behind behind the vaccine engine there's a there's a a gene
[01:21:50] engine there's a there's a a gene therapy engine where we we're trying to
[01:21:53] therapy engine where we we're trying to come up with way of expressing
[01:21:55] come up with way of expressing antibodies so the cell line development
[01:21:58] antibodies so the cell line development for commercial antibody is years uh from
[01:22:02] for commercial antibody is years uh from sequence in the RNA in your arm make it
[01:22:06] sequence in the RNA in your arm make it endogenously it it would be much faster
[01:22:08] endogenously it it would be much faster we could and so you know if there's a
[01:22:11] we could and so you know if there's a future pandemic we're thinking in The
[01:22:13] future pandemic we're thinking in The Hot Zone you're vaccinating with the
[01:22:16] Hot Zone you're vaccinating with the antibody system and then around the
[01:22:18] antibody system and then around the outside you're coming in with with with
[01:22:20] outside you're coming in with with with a vaccine uh to induce immunity and uh
[01:22:24] a vaccine uh to induce immunity and uh as a an a a therapy
[01:22:28] as a an a a therapy to challenge the those genetic therapies
[01:22:32] to challenge the those genetic therapies that we have now do you think that's
[01:22:34] that we have now do you think that's feasible for example we have a a crisper
[01:22:38] feasible for example we have a a crisper based genetic therapy that will uh
[01:22:42] based genetic therapy that will uh correct some some yeah if you
[01:22:45] correct some some yeah if you so crisper is a linear RNA speciality
[01:22:50] so crisper is a linear RNA speciality they need to be in Express done so it's
[01:22:53] they need to be in Express done so it's shorten up and you get your effect this
[01:22:57] shorten up and you get your effect this kind of self-replicating systems they're
[01:22:59] kind of self-replicating systems they're going to they're around for too long and
[01:23:01] going to they're around for too long and okay and I I the the gene editing
[01:23:05] okay and I I the the gene editing systems linear RNA with the guide uh
[01:23:09] systems linear RNA with the guide uh it's all about delivery they need to
[01:23:10] it's all about delivery they need to move away from the liver now and find
[01:23:12] move away from the liver now and find new targets and the deliver is the
[01:23:15] new targets and the deliver is the challenge and it's going to be a tough
[01:23:17] challenge and it's going to be a tough challenge yeah good thank
[01:23:22] you I think they asked my questions most
[01:23:26] you I think they asked my questions most but going back to the antibody therapy
[01:23:29] but going back to the antibody therapy so do you have a to have a some type of
[01:23:33] so do you have a to have a some type of attack that drives the RNA to B cells or
[01:23:37] attack that drives the RNA to B cells or you think that the any cells you put at
[01:23:40] you think that the any cells you put at RNA wood um the the early experiments
[01:23:44] RNA wood um the the early experiments are just I am injection and you know the
[01:23:47] are just I am injection and you know the muscle cells are able to produce lots of
[01:23:51] muscle cells are able to produce lots of antibod yeah and you know you code in
[01:23:52] antibod yeah and you know you code in the heavy light chain
[01:23:55] the heavy light chain the liver does a much better job so a
[01:23:57] the liver does a much better job so a lot of people have gone systemically and
[01:24:00] lot of people have gone systemically and and got production from the liver
[01:24:01] and got production from the liver they're better designed to produce
[01:24:03] they're better designed to produce antibodies you can measure high levels
[01:24:06] antibodies you can measure high levels of antibody in the they're not high
[01:24:08] of antibody in the they're not high enough yet so with some of the really
[01:24:11] enough yet so with some of the really potent antibodies serum concentrations
[01:24:14] potent antibodies serum concentrations can get up to levels of of efficacy
[01:24:17] can get up to levels of of efficacy uhhuh uh but it it's going to be 10
[01:24:20] uhhuh uh but it it's going to be 10 years of development uh uh we have to we
[01:24:25] years of development uh uh we have to we could probably get to the levels but
[01:24:27] could probably get to the levels but we've got to avoid immune stimulation
[01:24:30] we've got to avoid immune stimulation and generating antibodies against the
[01:24:33] and generating antibodies against the antibodies and the other question I had
[01:24:35] antibodies and the other question I had was the cost you already answered
[01:24:39] was the cost you already answered so what do you see what would be the
[01:24:43] so what do you see what would be the main challenge in terms of regulatory
[01:24:47] main challenge in terms of regulatory issues to approve the self
[01:24:51] issues to approve the self replicating yeah as a as a prod it's
[01:24:55] replicating yeah as a as a prod it's going to be perceived risks so um you
[01:24:58] going to be perceived risks so um you know the how long does the RNA persist
[01:25:01] know the how long does the RNA persist for which we can measure um what happens
[01:25:04] for which we can measure um what happens if is usually the what happens if you
[01:25:06] if is usually the what happens if you get a co- infection uh with an alpha
[01:25:10] get a co- infection uh with an alpha virus could they combine and so and
[01:25:14] virus could they combine and so and there are lots of studies showing that
[01:25:16] there are lots of studies showing that that that doesn't happen you can induce
[01:25:17] that that doesn't happen you can induce it in a Cell culture system where you
[01:25:20] it in a Cell culture system where you massively overload the system but they
[01:25:23] massively overload the system but they the alpha
[01:25:25] the alpha viruses super infection exclusion happen
[01:25:28] viruses super infection exclusion happen so one tends to replicate in the cell
[01:25:31] so one tends to replicate in the cell and not
[01:25:33] and not two so but we have to address them and
[01:25:36] two so but we have to address them and we have to generate data and certainly
[01:25:38] we have to generate data and certainly phase one everything went well but it's
[01:25:42] phase one everything went well but it's an ongoing dialogue with with The
[01:25:44] an ongoing dialogue with with The Regulators to to understand where and
[01:25:47] Regulators to to understand where and the public are going to react completely
[01:25:49] the public are going to react completely differently when they hear the word
[01:25:51] differently when they hear the word self-replicating and so we've got uh you
[01:25:54] self-replicating and so we've got uh you know educate them that this is a live
[01:25:56] know educate them that this is a live attenuated viral vaccine this this is a
[01:25:59] attenuated viral vaccine this this is a lot safer and why and convince them and
[01:26:02] lot safer and why and convince them and generate safety data and keep moving
[01:26:06] generate safety data and keep moving forward and it'll be a it'll take
[01:26:09] forward and it'll be a it'll take time we have a a uh question from the
[01:26:14] time we have a a uh question from the internet uh Daniel Duru he's asking if
[01:26:18] internet uh Daniel Duru he's asking if there is any limit to how much info in
[01:26:23] there is any limit to how much info in base pairs these uh vaccine constructs
[01:26:26] base pairs these uh vaccine constructs can can carry you said that
[01:26:29] can can carry you said that so there's no limit in terms of the
[01:26:33] so there's no limit in terms of the plasmid that we've seen yet but in terms
[01:26:35] plasmid that we've seen yet but in terms of
[01:26:36] of manufacturing uh at GMP compliant
[01:26:40] manufacturing uh at GMP compliant process where we have to have defined
[01:26:42] process where we have to have defined Integrity of full length and maintained
[01:26:45] Integrity of full length and maintained potency 16 KB is what we can do so
[01:26:48] potency 16 KB is what we can do so that's about uh 9 KB of coding uh
[01:26:53] that's about uh 9 KB of coding uh sequence which which is a lot there's a
[01:26:55] sequence which which is a lot there's a lot there's a lot we can do with that
[01:26:57] lot there's a lot we can do with that way more than what we are
[01:27:06] used Andrew um I was discussing with
[01:27:09] used Andrew um I was discussing with Sara if we use um nonm modify mRNA it's
[01:27:14] Sara if we use um nonm modify mRNA it's important to
[01:27:18] important to to to purify to remove the double strain
[01:27:21] to to purify to remove the double strain right because otherwise the immune
[01:27:23] right because otherwise the immune respones will be too strong right yeah
[01:27:27] respones will be too strong right yeah okay well so uh when you dose low it's
[01:27:32] okay well so uh when you dose low it's it's a dose dependent effect so so and
[01:27:35] it's a dose dependent effect so so and for vaccines it doesn't have to be zero
[01:27:38] for vaccines it doesn't have to be zero and so what what we
[01:27:41] and so what what we found is there's two ways three ways of
[01:27:44] found is there's two ways three ways of of achieving low double stranded RNA one
[01:27:47] of achieving low double stranded RNA one of them is you purify out I think you
[01:27:49] of them is you purify out I think you were some one of the students was
[01:27:51] were some one of the students was showing uh uh the fog
[01:27:54] showing uh uh the fog cellulose yeah uh you can some of the
[01:27:57] cellulose yeah uh you can some of the modern enzymes t7s have much much lower
[01:28:01] modern enzymes t7s have much much lower uh the yields are much lower though in
[01:28:04] uh the yields are much lower though in in the ivt reaction but I'm sure that'll
[01:28:06] in the ivt reaction but I'm sure that'll change the other one is you can if you
[01:28:08] change the other one is you can if you can measure it you can optimize the ivt
[01:28:12] can measure it you can optimize the ivt reaction so you can change temperatures
[01:28:14] reaction so you can change temperatures and times and concentrations of things
[01:28:17] and times and concentrations of things and the level of residual double
[01:28:19] and the level of residual double standard RNA will go up and down and so
[01:28:22] standard RNA will go up and down and so you can you can reduce it three ways
[01:28:24] you can you can reduce it three ways for me of the dosers we are it's easiest
[01:28:28] for me of the dosers we are it's easiest just to optimize the ivt reaction get it
[01:28:30] just to optimize the ivt reaction get it down to a level that uh is already
[01:28:34] down to a level that uh is already acceptable okay thanks and we're lucky
[01:28:36] acceptable okay thanks and we're lucky we have data where we know it's
[01:28:38] we have data where we know it's acceptable and and so with our assay and
[01:28:41] acceptable and and so with our assay and the problem is everyone has a different
[01:28:43] the problem is everyone has a different assay there's no standards yet uh but we
[01:28:47] assay there's no standards yet uh but we now as we switch programs we now know
[01:28:49] now as we switch programs we now know what residual levels are okay H and
[01:28:52] what residual levels are okay H and we'll just maintain those levels
[01:28:54] we'll just maintain those levels going forward okay
[01:28:58] going forward okay thanks okay um I think we we are good um
[01:29:04] thanks okay um I think we we are good um Andrew thank you very much again for
[01:29:07] Andrew thank you very much again for your generosity donating your time to be
[01:29:10] your generosity donating your time to be here three days five lectures uh we
[01:29:14] here three days five lectures uh we learned a lot and uh we hope that this
[01:29:17] learned a lot and uh we hope that this will be be just the beginning of
[01:29:20] will be be just the beginning of longlasting collaboration I do and we're
[01:29:22] longlasting collaboration I do and we're going to work together and conspire and
[01:29:25] going to work together and conspire and figure out how to make things happen I
[01:29:28] figure out how to make things happen I was overwhelmed by what you do here it's
[01:29:31] was overwhelmed by what you do here it's very impressive it's like end to endend
[01:29:33] very impressive it's like end to endend you do everything and uh with the new
[01:29:36] you do everything and uh with the new facility when you if you when you're
[01:29:39] facility when you if you when you're capable of GMP material going to phase
[01:29:41] capable of GMP material going to phase one it's going to be amazing yeah thank
[01:29:45] one it's going to be amazing yeah thank you thank you so much
[01:29:48] you thank you so much [Applause]

https://www.youtube.com/watch?v=cesLJwOePMk

Summary

TL;DR — Andrew Geall, a pioneer in mRNA vaccine delivery systems, discusses his extensive career journey and the development of self-amplifying RNA (saRNA) vaccines. He highlights the evolution of RNA vaccine technology, from early challenges to its current prominence, emphasizing the importance of scalable delivery systems and manufacturing processes.

Key points

• Geall's career spans over two decades, with significant contributions to drug delivery systems and mRNA vaccine platforms at companies like Novartis, Precision Nano Systems, and currently, Replicate Biosciences.

• He details the technical hurdles in developing RNA vaccines, including challenges in RNA synthesis, delivery via lipid nanoparticles (LNPs), and achieving competitive efficacy against conventional technologies.

• The presentation covers the journey of self-amplifying RNA (saRNA) vaccines, which offer potential advantages like lower doses and improved durability compared to linear mRNA.

• Geall showcases the rapid development and manufacturing capabilities for RNA vaccines, particularly highlighted during the COVID-19 pandemic, and discusses ongoing efforts to optimize vector design and production.

• He touches upon the potential of RNA technology for various applications beyond infectious diseases, including gene therapy and cancer vaccines, and the importance of antigen discovery.

Takeaway — The development of RNA vaccines is a complex, iterative process that has overcome significant scientific and manufacturing challenges, paving the way for potentially more effective and rapidly deployable vaccines and therapies.