The latest advancements driving neuroscience drug discovery research

DDW hosted webinar sponsored by ApconiX

In this webinar the following questions will be answered. Where are the breakthroughs in neuroscience research? How are advanced technologies proving beneficial to drug discovery and development? and how is the current unmet need within drug discovery and development for central nervous system, CNS disorders, providing opportunity?

You will hear from Dr Isaac Klein Chief scientific officer at Dewpoint Therapeutics, Dr Sam Clark CEO of Terran Biosciences and Professor Ruth Roberts Co-founder and Director of Safety Science at ApconiX.

Transcript of the Video

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[Music]

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Hello and welcome to this webinar titled ‘The latest advancements driving neuroscience drug

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discovery research’. My name is Megan Thomas and I’m the multimedia editor for

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DDW and your host for today’s webinar which is being sponsored by ApconiX.

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In this webinar the following questions will be answered. Where are the breakthroughs in

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neuroscience research? How are advanced technologies proving beneficial to drug

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discovery and development? and how is the current unmet need within drug discovery

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and development for central nervous system, CNS disorders, providing opportunity? You will hear

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from Dr Isaac Klein Chief scientific officer at Dewpoint Therapeutics, Dr Sam Clark CEO of Terran

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Biosciences and Professor Ruth Roberts Co-founder and Director of Safety Science at ApconiX. Over

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to you Isaac. Thank you for having me. My name is Isaac Klein, I’m the chief scientific officer at

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Dewpoint Therapeutics and I’m here today to tell you about biomolecular condensates, the Dewpoint

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drug discovery platform and how we can apply it for neuroscience drug discovery. So today I’ll

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give you a brief overview of condensate biology and how Dewpoint leverages our understanding of

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condensate biology to discover a new class of drugs. I’ll tell you a little bit about how

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abnormal condensates drive neuro degenerative diseases and the opportunity that gives us to

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find new molecules, that could potentially treat some of these difficult to treat entities and

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I’ll give you a snapshot of our ALS program, as well as our red syndrome program at Dewpoint. So,

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first a bit about condensate biology. The traditional view of the cell is that it is

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compartmentalized and organized by membrane bound organelles, things like the mitochondria, nucleus

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and golgi apparatus that we all know well. But over the last 15 years we’ve come to appreciate

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that there’s another level of organization in the cell, which is the condensate and these function

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as membraneless organelles, or biomolecules like protein and RNA co-ales and concentrate

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together in specified areas, to prosecute certain biochemical pathways or reactions.

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We now understand that these structures condensates regulate nearly every cellular

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process examined. There’s copious evidence that condensates regulate storage of certain entities

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in the cell, gene regulation is performed by condensation, both gene suppression as well as

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activation, signal transduction, nuclear import, signaling ribosome biogenesis on and on, are all

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cellular processes that are now understood to be regulated by condensates. The mechanisms by which

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condensates regulate these processes, shown on the right, are diverse and important for how those

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processes occur. Things like accelerating certain reactions and repressing reactions by sequestering

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biomolecules required for them into different compartments, as well as organizing cellular

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components such that reactions can take place in an efficient step-wise fashion. At Dewpoint, we use

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this new understanding to find condensate targets that are associated with certain diseases and

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drive those diseases and some examples are shown here. We have several examples, both internally

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at Dewpoint and broadly from the academic community studying this space, where condensates can drive

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oncogenic transcription that causes cancer.

We have examples where we find that abnormal

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condensates cause a toxic gain of function that

causes a cardiomyopathy. Many examples of abnormal

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condensates driving neuro degeneration, as well as condensates driving metabolic abnormalities like

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diabetes. Taken together, we find that these abnormal condensates or condensate opathies

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can drive many diseases and give us thousands of new targets to work on across therapeutic areas.

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We’ve also found that, once characterized, we can identify small molecules that address and

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correct these abnormal condensates that cause disease with profound therapeutic consequences.

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Some examples are shown here and the important point I’d like to make is, that the molecules

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that we discover against these condensates have entirely new mechanisms of action that could not

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be anticipated by traditional drug discovery approaches. In the dilated cardiomyopathy

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example I introduced on the last slide, where we’ve identified an abnormal condensate that

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causes a toxic gain of function and reduced heart function, we’ve discovered small molecules that

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dissolve these abnormal condensates and relieve that toxic gain of function conferred on the

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cell by the abnormal condensate. In colorectal cancer, we’ve identified a transcription factor

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that can be forced to condense upon exposure to a small molecule, thus sequestering it away from

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the genome and preventing anagen activation. In ALS, which is an example I’ll dive into a bit

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more deeply later in the talk, we’ve identified an abnormal cytoplasmic condensate that concentrates

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a factor from the nucleus and we can find small molecules that remove or departition that factor

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from the abnormal cytoplasmic structure and returning it to the nucleus of the motor neuron,

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thus restoring normal function. We’ve built a platform that is an end to end process for

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condensate identification and condensate modulator discovery and development, to explore this new

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space and take advantage of it to find new drugs for patients with diseases that have

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high unmet needs. We start with an identification platform component where we use human genetics to

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nominate new condensate targets in silico that drive specific diseases or pathogenic processes.

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We then model those condensates in disease relevant cell lines and using an inhouse

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platform and bespoke small molecule library, search for chemistry that changes that condensate

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in some desirable way. Using a battery of both condensate specific and functional assays,

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we then optimize those condensate modulators into drug-like molecules. We translate them

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in downstream models and then ultimately turn out drug candidates that can be brought to the clinic.

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The pipeline the companies develop using this platform is shown here. We’re primarily focused

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internally on oncology and neuro degeneration. Our lead program in colorectal cancer,

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which is looking at beta catene condensation, is set to hit the clinic at the end of 2025.

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Our next program is looking at disrupting mick condensation for treatment of ovarian cancer

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and that comes about a year later. In neuro degeneration, our pipeline is led by ALS, for

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which we’re selecting a development candidate now and targeting the clinic by the end of 2025. We’ve

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launched multiple collaborations with large pharma partners, with Bayer and cardiovascular disease,

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with whom we are prosecuting four programs in this space and with Novo Nordisk, in the diabetes and

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insulin resistance space, where we have multiple programs now in early discovery. In both cases,

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these companies have come to us and asked to use our platform to identify novel condensates that

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drive these diseases, as well as develop small molecules that address them for the first time.

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Briefly about neuro degeneration and condensates. This is a uniquely interesting space for the

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intersection of condensates and drug discovery. There are examples throughout the space of neuro

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degeneration, of condensates driving disease and some examples of that are shown here and it’s just

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a few. There are prominent examples, for example of condensates driving pathology in Alzheimer’s

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disease, in ALS, in several types of myotonic dystrophy, as well as other triplet expansion

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diseases like fuches retinopathy, condensates have been implicated in spinocerebellar ataxias,

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in Rett syndrome and many other neuro degenerative diseases as well. In an area, where there is a

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paucity of novel treatment approaches for these diseases with high unmet need, this is an exciting

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area to identify new targets as well as identify new molecules that can address these targets

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for the first time. At Dewpoint specifically, we’ve launched programs throughout this space.

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We have a lead program in ALS that I’ll tell you a bit about, as well as a program a bit further

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behind in Rett syndrome. We also have programs internally in myotonic dystrophy, Fewets retinopathy,

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as well as frontal temporal dementia. So a bit about our ALS program first. ALS is a difficult

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to treat disease and it’s caused by a seemingly desperate array of both genetic, environmental

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fact and environmental factors. What we’ve discovered is that nearly all cases of ALS, 97% of

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them regardless of the ideology of this disease, are caused by abnormal condensates that are formed

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in the cytoplasm of motor neurons and you can see those here. These condensates are specifically

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characterized by a concentration of a key splicing factor called tdp43, that in the disease state,

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is sequestered in these abnormal condensates,

as opposed to being in a nucleus where it should

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normally reside. This causes two things; one, there’s a toxic gain of function by inappropriate

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sequestration of this protein in the cytoplasm and two, there’s a loss of function because the

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protein is not where it should be in the nucleus. We modeled this in our laboratory and went to

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search for small molecules that move tdp43 out of the inappropriate condensate and back to the

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nucleus where it should be and that’s exactly what we found, this series of molecules that from which

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we are nominating a development candidate just in the next few months, the consequences of it are

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quite profound and shown on the next slide. First, across now 30 IPSC derived motor neuron models

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from patients with ALS, we can see a restoration of normal tdp43 function. We can see that in terms

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of normal splicing of tdp43 regulated genes like pole dip 3 and we can can see it in the expression

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of key ALS associated genes that are also driven

by tdp43 function, like statman 2. When we examine

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motor neuron survival as well as degeneration in the laboratory, we can see that our molecule

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condensate modulators is able to rescue neuro degeneration of IPSC drive motor neurons from

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ALS patients and remarkably in mouse model of ALS, we’re able to eliminate the expression

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of neurofilament light chain in the CSF of these mice. Neurofilament light chain is one

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of the only clinically validated biomarkers for ALS that is accepted by the FDA on for the basis

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of an approval of a new molecule. So this is particularly exciting as this has not been seen

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to our knowledge with any other therapies that are currently on the market. Briefly for Rett

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syndrome, I wanted to share some progress we’ve made in this disease. Rett syndrome is caused by

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an array of mutations in a protein called mecp2 but our platform nominated this is potentially

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being associated with an abnormal condensate behaviour. Indeed what we found, is when this

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protein mecp2 is mutated instead of localizing to the heterochromatin where it should be,

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it concentrates in a nucleolar condensate where it should not be, and this similar to ALS causes

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two things. One, is by abnormally localizing in the nucleolar condensate, it disrupts nucleolar

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function and by not being properly located in the heterochromatin compartment, it loses its ability

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to regulate normal gene expression. We modeled this in the laboratory and went to look for small

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molecules that reverse this condensate behavior and indeed we found chemistry that can pull the

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mutant mecp2 out of the nucleolus and restore its normal localization on the heterochromatin and the

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consequences of that molecule in cellular models are shown here. First on the left, you can see

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a normalization of mutant mecp2 localization out of the nucleolar compartment on the left. In the

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middle slide, you can see that not only is this mutant mecp2 removed from the nucleolus but it

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actually restores it, its localization in the heterochromatin is restored and on the right,

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you can see that nucleolar function is normalized upon exposure to this condensate modulator,

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as examined by phosphorylation of the S6 subunit of the ribosome which

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is a key measure of nucleolar function and health. Plenty more to see on our website.

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There’s a QR code on the left to the Dewpoint website, which has more information about our lead

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programs, our platform and our company and on the right, is a QR code that links to condensates.com,

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which is our educational website. More detail about condensate science, research, updates

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from our company in the field, as well as expert insights from those that study this broadly. Thank

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you. Thanks very much for that presentation, it was very enlightening. Next up, we’ll hear from

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Dr Sam Clark CEO of Terran Biosciences. Over to you Sam. All right, thanks. Today we’re going to

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talk about pro drug chemistry, breakthroughs in the schizophrenia space, focusing on how

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medicinal chemistry has been utilized to address unmet medical need in drug development and making

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better therapeutics in the neuroscience space. Now, first thing is how medicinal chemistry fits

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into a drug discovery engine. Typically when we think about drug discovery, we’re thinking

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about the development of molecules that have never been discovered before. The issue there is that it

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presents a high risk, in that most molecules are found to be toxic before they ever get into humans

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and then most don’t make it through phase one. So very few truly new molecules ever actually get

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into the target population with the disease. Then when they do, there’s the question of efficacy and

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that’s so why from such a large pool of molecules, so few make it through. There is a second way to

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address unmet medical need and that is through improving existing drugs, that have critical

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pharmacological issues that limit their efficacy. In this case, you have drugs that are potentially

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highly effective, could already address an unmet medical need but are not doing so due

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to pharmacokinetic limitations. What that means is that these molecules which may have been approved

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before, have problems getting absorbed into the body or metabolized creating either challenges for

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patient populations or limiting their efficacy, due to the way they’re metabolized in the body.

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Medicinal chemistry comes in, in it seeks to design and develop molecules that are targeted

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towards specific indications or development paths and, in this case, it involves designing, then

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synthesizing the molecules in the lab, putting them into animal studies and then eventual GMP

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scale up and human studies. So let’s talk about how that plays out. So a pro drug approach has

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been utilized to approve some of the leading anti psychotics on the market today and we’ll talk

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about the breakthroughs in schizophrenia around that. So first we need to cover what is a pro

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drug. Now pro drug is an inactive molecule that’s metabolized into an active form of the drug in the

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body and this molecule is engineered to provide advantages over the original drug to overcome

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pharmacokinetic limitations and let’s talk about how that’s been applied to the neuro psych space.

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Two of the most effective antipsychotics and most widely used on the market, are Aripiprazole which

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was Abilify and paliperidone which is Invega. Now, Abilify was originally invented as a once

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daily oral pill and it wasn’t until a number of years later that it was developed into a long

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acting injectable antipsychotic. That provides several benefits for patients in that patients

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with schizophrenia, often have poor medication compliance and when they are not taking the pills

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daily it can lead to reductions in the effective blood levels of the drug and a worsening of the

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disease. With diseases like schizophrenia we actually know that antipsychotics are disease

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modifying, patients that don’t receive sufficient doses of antipsychotics have a worse course of

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the disease over time and worse outcomes. So it’s very critical to have medications that

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have pharmacokinetics that work well with these patients and in this case Abilify, Aripripazol,

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was later improved via a pro- drug approach into this long acting injectable Aristata. Paliperidone

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similarly was also improved to make a long acting antipsychotic that first lasted 1 month,

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then the 3 month was developed and now they’ve developed the six-month injectable.

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So imagine instead of having to take pills every day, the patients just come in for a once,

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twice a year injection. So let’s just talk about that a little bit. The way they did that was,

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they attached pro drug moieties and what these are, are inactive esters on the side here with

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long carbon chains and this the palmitate esther here and this is the lauroxil esther here. Those

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provide lipophilicity to the molecules enabling them to be turned into long acting injectables and

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thus last a long time in the intramuscular Depot injection. There’s a second big advantage where

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pro drugs can address an unmet medical need and that is that they are able to take advantage of

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the FDA’s accelerated regulatory pathways. So for example, with Aristata, they were able to utilize

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the 505 B2 pathway and what that means is, instead of having to go all the way back to phase 2 phase,

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multiple phase three efficacy studies, which can take many years to get a drug developed and all

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the while patients are waiting, the FDA recognizes that pro drugs which are ultimately releasing the

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active molecule that was already shown to be safe

and effective. In this case, for Aripripazole laroxil, the

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active molecule had previously been approved as Abilify and so what the FDA requires for these

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improved versions, is pharmacokinetic bridging studies, to bridge the data to the old effective

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levels. This represents a major breakthrough for patients because patients with these pathways,

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you can get the drugs approved much faster with minor bridging studies, that are quick to do and

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often just one pivotal bridging study is all that’s required. In that sense, it’s a big

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win for patients because they don’t have to wait all of the extra years that are typically in play

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for a full phase 3 development program. Now let’s talk about one of the biggest breakthroughs right

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now in the schizophrenia space a new drug, first new mechanism in 50 years which is called KarXT,

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it’s a combination of these two molecules, xanomeline and trospium. Now the reason why this is such a big

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breakthrough is that xanomeline has a new mechanism of action, in fact it represents the first new

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mechanism of antipsychotics in over 50 years. All of the current antipsychotics on the market either

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directly affect the dopamine type 2 receptor and sometimes they also affect the serotonin 2A

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receptor and they work by blocking those receptors, but that mechanism has been rehashed over multiple

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different antipsychotics with different degrees of potency and binding over the last 50 years.

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Xanomeline acts on a completely different receptor system, the muscarinic receptor system, and in

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doing so that represents a big step forward, the first new way to treat psychosis and

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schizophrenia in half a century. However, xanomeline has some side effects because it activates the muscarinic

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receptors, which are beneficial in the brain for

patients with schizophrenia, it’s also activating them in

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the peripheral nervous system; in the stomach, in the intestines and activation of the muscarinic

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system there can cause problems like diarrhea, nausea, vomiting, sweating and so it’s been combined

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with another drug trospium. Trospium blocks the muscarinic receptors but it doesn’t cross the

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blood brain barrier and so the combination of the two represents a really unique mechanism where

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xanomeline is able to activate the receptors in the brain helping patients with schizophrenia. In trospium

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is able to block those receptors in the rest of

the body reducing side effects. Xanomeline has also

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previous been in trials where it’s shown efficacy in Alzheimer’s and other conditions so it really

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represents a paradigm shift for psych disorders. However, there is an issue with xanomeline and trospium

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and well first, before we’ll go into that, we’ll talk about that as part of the clinical studies.

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It’s been through three phase three clinical trials, shown a big reduction in schizophrenia

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symptoms and it’s up for potential approval

just next month in September. So very exciting.

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The downside is for all of these trials they have to use a twice daily oral formulation. Now most

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antipsychotics are at least once daily so to move to a twice daily represents a potential burden

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for patients. Now, with twice daily, the reason why is because the drug has issues with absorption and so

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the pharmacokinetic limitations are because these two molecules are very old, for example xanomeline

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was originally invented in the 1990s and trospium was invented in the 1960s, so as old molecules as

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you can imagine, they have not the pharmokinetics has not have not been improved since then. However,

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it still represents a big breakthrough and addressing an unmet medical need for a new

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mechanism that treats schizophrenia, without the side effects that typically are associated with

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classical anti-psychotics which can cause parkinsonian like symptoms in patients. Xanomeline and trospium don’t

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have those effects so it’s a big breakthrough for patients but it comes with a downside of a

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relatively, potentially difficult dosing paradigm, of having to take two pills a day every day to

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maintain efficacy and in a patient population like schizophrenia, where there are other

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options like long acting injectables, this poses a potential problem. Now, if we think about classic

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drug development such as you know developing an entirely new molecule, that could make patients

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wait a long time for a new therapeutic and it also comes with the potential for less efficacy. So far

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other muscarinic drugs in the space have not been able to reach the same efficacy and right now

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there are no other muscarinic close to approval but in clinical trials other muscarinic drugs, that

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have shown efficacy, have not matched the level of efficacy of xanomeline and trospium, the karXT

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efficacy. So, as you can see, sometimes developing a new drug isn’t the best option. So this is an

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example where medicinal chemistry can address that large unmet medical need and for example, creating

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pro drugs of xanomeline and trospium to fix the absorption issue and thus create a once daily form. This is where medicinal chemistry has been utilized in, for example, a terXT where they’ve

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created the decanoate pro drugs, here is what I’m showing you, where just similar to what we saw with

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Invega or Aristata, attaching fatty acids to the molecules it changes the properties and then once

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in the body the that body’s natural enzymes will cleave these, allowing normal xanomeline and trospium to

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produce its effects. That’s very exciting for patients because in that sense you can get the

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best of both worlds. You can get the better dosing regimen, the once daily oral and the long acting

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multi-month injectable, but also have the improved

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matched by any other more selective muscarinic drug in development. Just to illustrate

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what typically goes into a comprehensive prodrug program, these are very in-depth designed for terXT,

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involved more than 10,000 pro drugs, synthesizing over a thousand pro drugs with more than 200

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full-time chemists. Then in vitro studies, more than 500 of those and another 500 in vivo studies and

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PK modeling. So medicinal chemistry programs are no less intensive than classic novel drug discovery

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programs but they can result in enormous benefits for patients, cutting the weight time down by a

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large degree for patients waiting for new drugs and improving drugs that have efficacy and fixing

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the problems that may limit their use and that basically sums it up. Ready for any questions.

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Thank you for that presentation Sam. Next we’ll hear from Professor Ruth Roberts, Co-founder

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you Ruth. Thank you for the introduction and today

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I’m going to talk to you about iSLA, which is an integrated in vitro seizure liability assay which

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uses human induced pluripotent stem cell neurones or hiPSC and human ion channels. So let’s look

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at the problem we’re working on today. In drug discovery and drug development, as many of you

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will already know, safety is the main reason why drugs fail and you can look at this excellent

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paper from Cook et al in nature reviews, that looks at the AstraZeneca data set, showing that safety in beige

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brown there, is the main reason for failure. When we dig into the data further on in that paper,

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you can see that CNS arrowed here is top of the clinical failure table. So the main reason drugs

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fail in the clinic is due to CNS issues. It’s also an important reason for failure preclinically

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as we can see here and of those CNS failures, a significant number of those, are due to seizure

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which is unwanted electrical activity in the brain that crops up during the pre-clinical

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animal testing or even more devastatingly in clinical development as we can see here. So why

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does this happen? Well essentially our current approaches for screening for seizure don’t work

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and this is a rough schematic adapted from Alison Easter’s paper from 2009, where we can see during

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the important GLP toxicology phase in the center of the screen, that precedes first time in humans,

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we conduct a standard regulatory package. Within that, we can also look at rodent behavior and EEG

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to confirm any seizure type activities that may have been seen in the animal testing in GLP tox

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and back in 2009, Alison’s paper was suggesting that we could screen to prevent these seizures

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as a pre-screen by using rat blown brain slicers or perhaps zebra fish assays, to avoid going

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into animal testing with compounds that had the potential to cause these seizure like activities.

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Those tests were introduced but in my view they haven’t really made that much of a difference to

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the failure rate as you can see. So we really need to improve preclinical seizure screening

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because essentially it currently, it’s very hard to detect, the current methods are very low throughput,

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they’re expensive and if they’re done in rats and dogs or in rat brain slices or zebra fish, we have

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poor translation to humans. So we thought it was a good time to rethink and redesign our approach to

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screening for seizure. How did we do this? Well we started by putting an idea to an SOT workshop way

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back in 2019 co-chaired by Jennifer Pearson from HESI and described in our paper from 2019.

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We put to this committee, or this workshop, could we actually approach screening for seizure based

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on human cellular models and human ion channels and this was discussed at this workshop in great

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detail and subsequently published in these papers you see here. What we put to this workshop was, we

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could screen for seizure using a micro electrode array abbreviated to MEA. This describes the MEA

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assay, so on the left hand side you’re looking at neurons plated in the micro electrode array plates

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shown here and each plate has a number of wells and each well has 16 electrodes in the bottom of

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the well. We then look at electrical traces shown here, from each of the 16 electrodes numbered 1

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to 16 on the day five pattern and over the time course between day five to day 23, these plated

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human neurons settle down into a nice harmonous, peaceful firing pattern as you can see in day

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  1. So they need to make connections in order to behave like normal neurons but by day 23, we’re

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seeing a nice normal firing pattern. We then tested these neurons with a variety of compounds known to

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cause seizure. shown on the left hand side. So as you can see in this graph, we’re looking at the

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alterations in the micro electro parameters with the number of seizurogenic compounds compared to,

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at the bottom the negative control paracetamol or acetamenophen. We see a whole different range

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of impacts on the MEA parameters with compounds known to cause seizure. So essentially in this

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human neuronal assay in vitro, the majority of seizuregenic compounds increase network burst frequencies,

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decreased duration and decrease the number of spikes per network bursts. So we’re able to

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see the phenotypic manifestation of seizure in this human invitro

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assay. As well as looking at a phenotypic assay, which is human neurons, we wanted to also look at ion

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channels because we know they’re associated with seizure. So we know there are variety of

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epilepsies there are human ion channel mutations, so we listed all of the potential ion channels

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that could be associated with seizure and as you can see at the bottom, we started with hundreds

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of ion channels and then based on expression profile literature on their function and the

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pharmacology associated with them, we narrowed it down to 15 seizure related ion channels. This is

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the same compound you saw on the MEA data across the top but down the left hand side are our 15

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ion channels associated with seizure. What you can can see here, is with CNS active therapies

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associated with seizure particularly like, for example amoxapine chlorpromazine, you see a high

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impact in other words inhibition at under 30 micromolar of signaling at these ion channels

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which is associated with seizure. So we now have a really elegant assay where we’re able to see the

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seizurogenic phenotype and induce human pluripotent stem cell neurons in vitro and then we can look at

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the mechanistic basis of that seizure, looking at the different ion channels implicated in seizure

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and looking at the profile of each compound against those ion channels. So we published

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this work back in 2023, where we described this integrated approach. All the data I’ve shared

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with you today are in this paper. Please feel free to take a look or email me and I can send you the

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paper and we were very excited, on the back of this paper, when it was mentioned in a publication from

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the FDA, where they were looking at new approach methodologies in the nonclinical assessment of

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pharmaceuticals and our paper is cited in this paper from FDA, Avira et al, as a new approach methodology

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with a context of use around screening for seizure. So please take a look at that paper too, it’s very

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interesting reading. So what we’re saying is, that iSLA, the invitro seizure liability assay, is an

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innovative new approach to screen for seizure and essentially what we’re using it to do, is to

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replace the in vitro or ex vivo models here at brain slice and zebra fish, to screen very early

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for the compound’s ability to potentially cause seizure, but can also be used if seizures

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are seen in animal studies in the GLP tox phase or in humans in the clinic, to go back and look

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at the mechanistic basis of those potential seizures, looking at ion channels. So we’re really

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saying that this essay has got multiple uses in understanding seizure and it really is a complete

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redesign of our thinking. We’re not trying to replace a particular animal test with an invitro

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assay. We’re saying let’s step back, look at what we’re trying to do to ensure patient and volunteer

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safety and then design something that’s human specific, that can be translated directly to human

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risk benefit in the clinic. So how is iSLA used? As I’ve already alluded to, there are two main modes

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of use of iSLA. The first is the proactive which is the predictive, in other words we’re looking

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at MEAs for a chemical series to prioritize which is the more safe series regarding seizure or early

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hazard identification and redesigning compounds that may have a seizurogenic potential, via

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interaction at an ion channel and then later on,

as I already alluded to, when issues have been seen

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either in the animal tests or in clinical trials, we can do an MEA study to understand the human

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relevance of the seizures seen in animals and an ion channel panel to then understand the molecular

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basis. I have a couple of case studies to share with you which are very recent data of work

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we’ve been doing with clients. In this example, there was a very difficult situation where the

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compound caused convulsions in the dog but not in other preclinical species. So was the convulsion

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seen in the dog relevant for humans is of course the really important question. When we looked at

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this on the MEA assay, there was a dog specific metabolite that caused seizurogenic phenotype. This

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metabolite was not present in the rat and is not present in humans, so it looks like this is dog

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specific. Then when we went to an ion channel panel, we saw a hit on an ion channel panel that

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gave us mechanistic insight into the molecular basis of the dog specific seizures that were seen with

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this compound and this data accompanied the IND, when that was submitted shown here. In the second

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example, there was seizure seen with a chemical series and the challenge here was, we need to

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identify a chemical series with low seizurogenic risk, so we see here chemical series through A, B, C, D

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and E and when we looked in MEA, we can see here that series A had a low risk whereas series E had

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a high risk and then a smattering of scores in between the two. So we guided this client towards

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series A, which had distinct structural features from the other compounds being considered in lead

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optimization. So we were able to design from the very beginning, a compound with low seizurogenic risk

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compared to those that may have a risk in either preclinical or clinical development,

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which is a really great outcome and saves a lot of time and effort later on, trying to understand

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issues that can be avoided. So to conclude what I’ve shared with you today, seizures remain an

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issue in preclinical and clinical development. So we still see a high incidence of seizures

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in animal tests in the geop animal studies and then also in the clinic. iSLA uses human induced

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pluripotent stem cell neurons and micro electrode array in a bespoke ion channel pane,l to detect

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and understand the molecular basis of seizures when they occur or to predict and prevent them. We

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have great translation to humans because these are human cells and human ion channels. As well, we

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have really great three R’s benefits so it’s been an interesting discussion, I have been asked which

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test are we replacing? Are we reducing animal use or are we replacing animal use? Yes we are in a sense,

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that only those compounds that go forward into GLP toxicology testing in animals, have only

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those compounds that have been already screened for seizure liability will progress so we’re not

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wasting valuable resources and time testing compounds that we could have predicted, would

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not make it through to the clinic. As I’ve already described, this work has been used for the

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proactive mode which is the ideal place to be, to design compounds, select between the lead series and

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optimized molecules and then, in problem solving where we are looking to explain observed

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seizures and their mechanistic basis. Finally I’d like to acknowledge the team who were behind this

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work. Here we see the whole ApconiX team at a recent science day, top left, particularly I’d like to

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acknowledge the ion channel lab and MEA team led by Mike Morton. The work was done primarily by

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Kim Rockley, shown bottom left and also the rest of the lab team; Karen, Emily, Magalie,

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Hannah, Beth, Katie and Louisa, all of whom have contributed to this extensively. I’d also like

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to acknowledge our collaborators Bristol Meers Squib, who provided some of the rat brain slice

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data for our comparisons and Axion Biosciences, who provided the maestro machine that the MEA

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recordings were done in and Fujifilm Cellular Dynamics for the human induced pluripotent stem

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cell neurones. And finally, just a happy to say we won the Kings Award recently. So those lovely

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pictures top right, is me smiling away at Windsor Castle going into receive The King’s Award for

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the great work we’ve done at ApconiX so thank you. Thank you for that presentation Ruth and

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thank you to everyone who has presented today. Before we continue to the Q&A,

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I’d just like to remind everyone that this webinar will be available to access on demand on the DDW

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website www.ddw-online.com. We’ll now move on to the questions. Isaac, this first question is

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for you. Can the same condensate drive more than one disease, can cmods developed for a particular

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target be used in other diseases? This is one of the reasons we’re excited about condensates

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for drug discovery broadly and ALS actually provides a really nice example of this. So

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abnormal TDP 43 condensates in the cytoplasm, well established to dry almost 100% of ALS cases, but we

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also now understand that it drives about half cases of frontal temporal dementia and may also

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be associated and drive other neurodegenerative diseases, like chronic traumatic brain injury and

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even a certain percentage of patients diagnosed with Alzheimer’s. So we’ve already shown in

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cellular models, that our molecule can be potentially used to treat frontal temporal

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dementia. We’re exploring other indications now and this pattern of one condensate driving multiple

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diseases is also seen in the oncology space and so short answer is, yes a molecule discovered

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by our platform for a particular condensate, could potentially be applied to other diseases

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driven by the same structure. Thanks for that answer. Isaac, the next question I have here is,

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can condensates be used as biomarkers and can they be used to identify patient populations that would

00:42:26,000 –> 00:42:33,520

benefit from treatment with a specific cmod? Yes, so one of the many advantages of condensate based

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drug discovery, is that they’re highly translatable entities. ALS here provides another outstanding

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example, where the same condensate abnormality can be seen in cell line models, animal models as well

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as in patient biopsies. So that provides us a nice chain of translatability all the way from

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discovery to the patient. In terms of biomarker use, that is certainly an area that we’re excited

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about. We’ve already demonstrated that condensates can be used as biomarkers or specifically in terms

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of patient selection, both in the neurodegenerative space as well as in the oncology space. Also

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importantly the downstream consequences of abnormal condensate function, once understood, can

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also be used as biomarkers for patient selection and potentially even to monitor the response to

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therapy with a condensate modulator. That was a great answer Isaac thank you. The last question

00:43:28,240 –> 00:43:33,880

I have for you here is, what role do condensates play in more common neurodegenerative diseases

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like Alzheimer’s or Parkinson’s? Could the Dewpoint platform address these needs? Absolutely,

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they do play a role in those more common diseases like Alzheimer’s disease, Parkinson’s disease and

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others. We simply haven’t tackled them because there are areas that aren’t quite bite-sized

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for a small biotech to tackle from a development standpoint but we’re excited to, once we reach that

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stage and certainly if there were any partners, that are interested in leveraging our platform

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to find and develop small molecules for those larger indications, we’d be very excited to tackle

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them sooner than we had anticipated. That’s great Isaac thank you for that answer. Sam I now have a

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question for you. How does a pro drug approach to improving existing therapeutics compare to

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designing new therapeutic molecules? So when you take a pro drug approach you’re very much focused

00:44:29,280 –> 00:44:36,520

on one target that you already know. So you have a drug that you already know works and you already

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know that drug is safe, so those are two massive benefits. There are big benefits for patients

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in addressing an unmet medical need. In a classic drug discovery program you’re starting with an

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unknown. Perhaps you used modelling to try to design a number of new structures or classically you know

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had people come and design them those by hand. But either way, you’re working with new structures that

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have never been tested in humans, have never been shown to be safe and more than 90% get knocked out

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before phase one and/or post phase one and then by then, you still have the efficacy issue. Even

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if they’re found to eventually be safe and most aren’t, then the question of whether they’ll be

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effective is a big question. That’s what we’ve seen with the newer muscarinic, this a normally was

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invented in the 1990s but it’s still, to date, the most effective muscarinic in clinical trials so

00:45:30,440 –> 00:45:36,440

far to treat schizophrenia. The newer ones developed since then, in the 2000s and the last

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decade, have not been able to match the efficacy of the original. So with medicinal chemistry you start

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with the molecule you know is safe and effective and then you just fix a few problems on it to

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improve its pharmacokinetics, its absorption and potentially even reduce side effects. So it’s very

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exciting by starting with a known component. However similar to classic drug development, it still can

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be extremely in-depth program involving thousands of designs. Thanks for that answer Sam. The next

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question I have here for you is, how would changing the pharmacokinetics of previously

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approved therapeutics contribute to addressing the unmet medical need for CNS disorders? So

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there’s actually a couple ways. One of which that we talked about earlier in the talk, was

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simply through improving the dosing paradigm. With patients like schizophrenia or patients with

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Alzheimer’s disease, taking pills multiple times a day represents a burden and a potential for

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where these the patients may not be compliant with the medications and thus not get the amount

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of the drug that’s effective. With a pro drug approach, you can improve the pharmacokinetics

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and thus improve the experience for patients and ease of dosing and thus the potential outcome of

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taking that medication. A second way, though that we haven’t gotten into as much, is that you can

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potentially reduce side effects by changing the pharmacokinetics. Many side effects of drugs are

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often of immediate release drugs the ones that are not on an extended release, are often due

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to the spikes in plasma concentrations due to the immediate effect of the drug. Now with a pro

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drug approach enabling longer acting formulations, it smooths out the pharmacokinetic profile of the

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drug, reducing plasma spikes that could cause side effects and thus potentially improving things like

00:47:38,680 –> 00:47:45,400

nausea, vomiting and diarrhea and other spike related incidences, with a nice smooth long-

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acting profile. It further addressing the unmet medical need by providing patients with drugs

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with better safety profiles as well. So in this sense, we have two ways that medicinal chemistry

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and thus the pro drug approach has been used to really address this unmet medical need, with on a

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fast timeline using the FDA’s accelerated pathway and pro drugs now have been used to improve many

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neuroscience drugs to date, such as, for example, a Gabapentin, other ones and as well as,

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drugs in the antibiotic space and across all the different systems. So it’s a widely used strategy

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that results in big breakthroughs for patients. Thanks Sam. A final question for you here. What

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role does medicinal chemistry play in advancing drug discovery? With medicinal chemistry it’s

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not just making small modifications, it those modifications that shed a lot of light on structure

00:48:48,920 –> 00:48:54,960

activity relationship and that’s help push the field forward in the development of understanding

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how structural modifications to molecules, can create large improvements in those molecules.

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So, in addition medicinal chemistry often utilizes a lot of novel synthetic roots

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to make these molecules in a way that would be safe for human consumption and often GMP

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processes and so medicinal chemistry in addition to improving the molecules is also pushing forward

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the chemical field and our depth of what’s possible in chemistry because there may be

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molecules out there that could potentially be improved by pro drugs but no one knows

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how because they’re considered difficult molecules. With new approaches in medicinal

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chemistry’ been able to improve upon that and thus widen in the space and this has

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been utilized in neuroscience, as well as many other systems now to improve a lot of existing

00:49:47,240 –> 00:49:52,400

molecules and and even push through classic drug discovery based on the discoveries from medicinal

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chemistry. Great thanks for those answer Sam. The next question I have here is for you Ruth.

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Why is CNS safety such a concern? Yes, that’s a really great question. CNS safety is a big

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concern for two reasons. One is because it’s the primary reason for clinical failure and having

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a drug fail by the time it gets to the clinic

is really bad because you’ve invested so much

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resource by that time. The other thing is one of the simple ways that people say to avoid

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CNS liability or CNS issues, is to avoid blood brain barrier penetration. Which is fine if your

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compound isn’t intended to get into the brain but for CNS therapeutics of which many people

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are involved in delivering and developing CNS therapeutics for anxiety, for Parkinson’s,

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for depression etc, these compounds need to get into the brain so we need to understand early

00:50:56,800 –> 00:51:04,120

a mechanistic level how to distinguish the efficacy from the unwanted side effects at

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seizure related ion channels. Thanks for that answer Ruth. The next question I have for you is,

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is this assay designed only for CNS targeting? Yes, so again I touched on that before, is that

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this assay is vital for anyone developing CNS targeting therapeutics because they are

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intended to penetrate the blood brain barrier and therefore, if they have any interaction with these

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ion channels, they are likely to be associated with a CNS risk. However, if you look back at

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the AstraZeneca data set, although CNS targeting therapies are a hot spot for CNS toxicities as

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you would expect, there are also many compounds in other therapeutic areas such as oncology,

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rheumatoid arthritis, anti-infectives, that also have a seizurogenic risk and that’s presumably

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because in order to get the efficacy required, those compounds happened to have an interaction

00:52:03,800 –> 00:52:08,840

with ion channels in the brain. But knowing now what we know about ion channels and their

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associated with seizure, we can work to design that out early not just for CNS therapeutics but

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for other therapeutics as well. Thanks Ruth and a last question for you. Why are current methods not

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able to detect these issues? I think the primary reason is translation. So we are dependent on two

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species toxicology testing in the GLP phase, which would more often than not be rat and dog.

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It would seem that the seizurogenic mechanisms do not translate from the rat or the dog into humans.

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So we can’t assume that testing in those tox species will detect human risk. Having said that,

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we do have some interesting in vitro models of course, zebra fish and rat brain slicers, that

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are not using whole animals but again there’s a translation challenge. So I think it’s primarily

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around translation and some of the tests in the past have aim to replace whole animal studies with

00:53:14,400 –> 00:53:21,280

maybe in vitro tests done on zebra fish or rat cells. So we essentially have gone back to basics

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and said this needs to be a human-based system if we’re going to improve at help’s testing so again

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I think the primary reason is lack of translation to humans. That brings us to the end of this

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webinar. I’d like to thank everyone for tuning in and a big thanks to our presenters. If you would

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like to listen to the webinar again it will be available on demand at www.ddw-online.com. Thank

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00:53:50,320 –> 00:54:00,240

you for the sponsor of today’s webinar ApconiX and thank you to the audience for listening.