0:00
Smaller particles, so these are very small particles, 20 nanometres, not 2 millimetres, so about 10,000 times smaller I guess.
0:06
So when we talk about Cytolytix and some formulation aspects around this programme that we've got, it's a next generation oncolytic peptide.
0:14
And so main focus is in cancer therapy.
0:17
We're listed.
0:18
So we've got the usual stuff.
0:20
So just top level in terms of the company.
0:22
So I'm the CEO of ValiRx.
0:23
ValiRx is a company that basically screens for early stage assets.
0:27
So we typically look for assets from preclinical in tech transfer or small biotech.
0:33
If we like them, we evaluate them.
0:34
If we still like them, we bring them in house and we set them up in a special purpose vehicle.
0:38
So Cytolytix is one of our special purpose vehicles.
0:40
I'll explain that a little more, but you know, our kind of job is to kind of identify early stage assets, take them through this evaluation and then licence some kind of pathway to de risk assets taking for taking them forward.
0:53
So we're looking to basically develop assets through to IND ready and then try and partner them at that later stage.
0:57
So we've got some examples here of companies and universities that we've been working with across the board going forward.
1:07
It's a joint venture.
1:08
So Cytolytix is a joint venture with the academic institution that we partner with King's College London.
1:14
We've been basically developing this membrane disrupting peptide that was invented at King's College London.
1:20
It's got a high degree of cancer selectivity.
1:22
It's really interesting.
1:22
So you know, there are oncolytic peptides in the markets, but one of the key features of this one, we see a 200 fold differential activity between healthy cells and cancer cells, which is important for therapeutic index.
1:36
There's also the potential to activate the host immune system and that's a key part of this.
1:40
So you can basically activate the host immune system by releasing neoantigens from the cells that you kill.
1:46
And then you can get kind of an abscopal effect where you can basically see a broader systemic activity and also for synthetic lethality.
1:52
So if you can punch holes in cells, you can let small molecules in there as well.
1:56
So that's the interesting co-formulation.
1:58
We're looking at applications where we can use small molecule drugs in combination with the peptide.
2:04
Our primary focus is on triple negative breast cancer.
2:06
So it's a subset of the broader breast cancer market, but highly metastatic, highly aggressive and real high unmet clinical need.
2:15
So as a company we're focused generally on Women's Health and oncology so that it's really keeping with that ethos market slide here.
2:23
But basically you know, it's a large market triple negative breast cancer.
2:27
All of the assets that we look at including this one though have pan cancer potential.
2:30
So we're not just limiting ourselves to a single indication.
2:34
So it opens up kind of partnership opportunities around immune oncology space, small molecule space.
2:40
I'm actually very interested in ovarian cancer as well.
2:42
So we have some prospective collections underway for ovarian patient derived cell models at the moment that allows us then to bring in combinations with the immune system because we can collect blood samples alongside those primary biopsies, and you can look at combinations there.
2:57
We're also interested in prostate cancer, and we have a collaboration with the Open University in the UK, particular expertise in neuroendocrine prostate cancer, which is again a highly metastatic, highly aggressive disease.
3:08
You can see the kind of the space that we're playing in with the PD-L1 inhibitors in the combination therapies with immune oncology, it's very attractive.
3:16
So just a top level where we are with the technology, how it works.
3:19
So it's a nano therapeutic, as I said, so very small particles and that's important from the ability to target into solid tumours.
3:26
But there's some molecular simulation design that went into this basically.
3:31
And you can see that what we've essentially done is taken this peptide, it's a 15 amino acid peptide built around essentially host defence peptide.
3:38
So these are designed to essentially to protect you against bacterial infection.
3:42
We've got this section that forms an alpha Helix, or it wants to form an alpha Helix.
3:46
So it's not quite formed an alpha Helix, but what happens is when it interacts with the membrane of a cell, it basically creates a hydrophobic environment which drives this alpha Helix formation.
3:58
And we've built a library.
3:59
So there's a base peptide of 14-15 amino acids and then you switch out selectively different amino acids within that.
4:05
And that's the screening programme.
4:07
So we use molecular dynamic simulations to optimise the sequence.
4:10
It's a rational design.
4:12
They're all natural amino acids, which again is important for certain applications.
4:15
I'll touch on where you might want to express that peptide rather than synthesise it.
4:21
We screened using a range of models, so 2D and 3D organoid, spheroid type models.
4:27
And again, the characterisation in silico is really important because you can see basically you can visualise these pores that form.
4:34
So it's important also to remember that the peptide itself is zwitterionic.
4:37
A lot of these host defence peptides tend to be cationic.
4:40
So you lose the selectivity there because you get this nonspecific interaction with cells.
4:44
So the zwitterionic nature of the peptide is a key part of its selectivity and it also means that as you drop pH in the low extracellular environment and also within the endosomal compartment, you then you start to see pH driven activation as well.
4:56
So the key part and the reason that we're at the meeting here is to talk about the nanoformulation.
5:01
So it's nano stabilised.
5:02
So that allows you to then think about systemic delivery.
5:05
So again, there's a couple of peptides on the in clinical development at the moment, but they're administered intratumorally because of the systemic toxicity by formulating within a nanoparticle, then it opens up options then for different types of delivery.
5:17
And it also allows you to think about PK benefits and targeting where the nanoparticle goes rather than what the peptide itself is doing.
5:24
So you can change the kind of route of administration, you get local concentration effects as well.
5:30
And yeah, so basically to IV routes of administration.
5:33
We've got some nice mouse data showing that we could administer these by IV.
5:37
So this is just a representation of the peptide.
5:40
You can see the key features here.
5:41
So the EEK relates to the kind of the glutamic acid residues here and then the lysine residue on the end here.
5:48
So those are the bits that we've switched out, the differential bits that build up the peptide.
5:52
There's a tryptophan on the end that's really there just to provide a UV absorber that you can use for quantification.
5:59
So it's 15 amino acids, it's all natural nanoformulation, and then there's just really nice molecular dynamic simulation work.
6:07
This was done by the academic founders.
6:09
We're basically showing this principle where you've got the interaction initially with the membrane, so individual unstructured kind of peptides that interact with the membrane.
6:17
They basically assemble on the membrane and then you get 8 of them that make this pore.
6:20
So viewed from the top and from the side, you get this pore in the membrane.
6:24
Cells don't like holes in them.
6:26
So that basically, you know, kills a cell, allows neoantigens out and also is a portal then for small molecules going in.
6:33
So just some data showing the effect here.
6:35
So this is a triple negative breast cancer cell line, and you can see across the top in grey, these are the controlled nanoparticles.
6:42
So just PLGA nanoparticles on their own, completely innocuous.
6:46
The peptide on its own basically has this kind of IC50 of around 60 micromolar, which sounds high, but it's typical for these types of these peptides.
6:56
But what you see when you basically formulate it into the nanoparticle, you get a shift in terms of the IC50.
7:02
So it's already 4 times more active.
7:04
And we think that's because of basically this concentration effect.
7:06
You need eight of these peptides in order to form a pore.
7:09
So by bringing them together on the nanoparticle, and I'll show you an image of this, you can actually present them in a more efficacious way.
7:17
So here's the picture.
7:18
These are the nanoparticles.
7:19
So you can basically see that it's a solid.
7:25
Now it's a solid nanoparticle, but there is a small core, there's a hole in the centre.
7:34
So we think this peptide in the centre is not active because the nanoparticle would have to dissolve in order to release and the release profile from PLGA is in the order of weeks and months.
7:45
So at the nanoparticle level, what we're seeing is the PEG on the outside, a solid hydrophobic core of PLGA and then the peptide that sits on the surface is basically in the active form.
7:55
So what we think is happening is an exchange from the surface of the particle into the membrane of the cell, another key piece as well.
8:04
So these peptides, as I said, you don't want them interacting with cells non-specifically.
8:10
And a key metric is the ability of the cells not cause a haemolysis.
8:16
So rising the red blood cells.
8:18
And you can see the free peptide on its own does have a, about a 20% level of haemolysis here.
8:24
But in the nanoparticle, you can bring that back down to base level.
8:26
So that means you can systemically administer these in a safe form.
8:31
And then there's just an example of the mouse work that we've done here.
8:34
So you can see basically that you get really nice control of the tumour size in a mouse model after treatment, whereas in the controlled nanoparticles you get essentially uncontrolled growth of the tumour.
8:48
So just one word in terms of the uptake.
8:50
So the free peptide we think is active on the extracellular membrane of the cell.
8:54
In a nanoparticle format, it does change the route of administration either by passive uptake into the cell through phagosomes or pinocytosis into these vesicles or in fact receptor mediated endocytosis which can trigger accelerated uptake but also position the peptide nanoparticles at the kind of interior surface.
9:15
So at the moment everything we're doing is non targeted, but we have got programme under way with a group with a HER2 targeting aptamer.
9:23
And we think that will again enhance both the uptake speed but also present the peptide at the interior surface of the vesicles, which could contribute then to an increased level of activity and push that IC50 even lower.
9:38
So just in terms of overview, the formulation is the key challenge, but it does present opportunities for partnering.
9:45
And we've been talking to a number of people at this meeting about where we go forward with that.
9:50
We've got efficacy and safety testing under way with a range of different formulations.
9:53
And I'll show you those.
9:55
The idea is that we can pick the best formulation, and it may be that it's a different formulation for depending on the route of administration.
10:02
So you know, 1 route of administration may be benefit from a different type of formulation.
10:08
And we've got this ovarian, sorry, the prostate cancer target expansion underway working with the Open University.
10:13
It's got multi cancer potential and we're looking to exploit these three partnerships going forward.
10:18
So the key for the primary formulation that we're looking at is this Poly Lactic-co-Glycolic pegylated version.
10:24
And that was the original, the formulation that was developed by Kings.
10:27
We've got our own in house liposomal formulation now.
10:30
And that's interesting because we think the lipid helps to form the alpha helix.
10:35
So it's already in a pre-activated form.
10:37
So there's some advantages there.
10:40
We've got a Canadian company, hence the Canadian tag that we've been working with on a micelle format.
10:45
So this is a spontaneously forming kind of micelle format, which again we're in the process of developing that at the moment.
10:51
So we'll know if that has advantages in terms of efficacy.
10:53
But then the novel system here, this is actually a virus.
10:56
So Omios is a San Francisco based company with an oncolytic virus and they're looking at basically incorporating the peptide into the virus to enhance its ability.
11:06
So the virus itself is active, it's oncolytically active, but it doesn't escape from the cells exceptionally well.
11:12
So the idea is that we use the pore formation to allow the virus to escape locally and be more efficacious.
11:20
So just in terms of how we make these things at the lab scale, it's relatively simple.
11:23
This is a slight oversimplification, but essentially you've got a peptide and PLGA dissolved into an aqueous miscible organic solvent.
11:31
Drop it into an aqueous solvent, you get solvent extraction and spontaneous formation of nanoparticles, which you can see up here, really nice tight control.
11:39
So as I said, they're about 20 nanometres.
11:41
And then the just looking at the Zeta potential here as well.
11:44
So you know, the Zeta potential reflects the nanoparticle itself, not so much the peptide on the surface of it is protected.
11:50
And you can see that you've got this kind of opalescent kind of view.
11:53
One of the key formulation issues with these nanoparticles is they do tend to aggregate.
11:57
So we spend a lot of time looking at how we do that stabilisation.
12:00
And there are some tricks to that.
12:02
I'm not going to say what those are because they're proprietary, but we've got a stabilised formulation, freeze dried now.
12:12
So particular stabiliser that we've got in there, you need to do solvent extraction to get rid of that organic solvent.
12:19
And then stabilisation's relatively straight forward in terms of the lyophilisation.
12:23
You just use sucrose as a cryoprotectant.
12:25
So the key features that we're looking at are things like particle size, particle stability, Zeta potential, encapsulation efficiency, which is a key metric really how much peptide do we have in each of those nanoparticles?
12:36
And we have a protocol now it's completely scalable.
12:38
So I promised I'd give these guys a shout out Unchained.
12:41
They've got a booth in the main hall 59, I believe it is.
12:46
And they've been working with us with their, with one of their lipid formulation systems also works very nicely with nanoparticles.
12:53
So you can use these kind of high efficiency mixers, speeds things up, it's really reproducible.
12:57
It gets you much better encapsulation efficiency as well.
13:01
So making the particles themselves is relatively straightforward.
13:03
It's driving that nanoparticle loading, which is the key in the reproducibility around that.
13:11
So, yeah, essentially it works by using these tangential mixers in a chip format.
13:16
You get really nice assembly, and you can see that using different percentages of stabiliser here, you can basically reduce the size, and you get to a kind of a critical size that's completely stable.
13:28
And yeah, just an image of the mixing device here.
13:32
So then last slide, I just wanted to talk about the immune activation component because I think this is really exciting for us, the ability to engage the host immune system.
13:40
So you see a big amplification effect in terms of the overall efficacy of the peptide itself, but also a more sustained effect, right.
13:51
So ultimately what you get here is essentially a vaccination approach.
13:54
So you get local cell killing release of neoantigens, particularly, this is important for things like prostate cancer, for example, which is historically it's a kind of biologically, it's a kind of a cold tumour from the immune perspective.
14:05
So treating with these types of activation systems.
14:08
This LTX-315 is a, is another oncolytic peptide.
14:13
We've been tracking these guys.
14:14
They've got an intratumoral clinical trial underway for melanoma and they see that basically their peptide basically works in a very similar way.
14:23
It'll kill cells, presents neoantigens basically that are recognised by dendritic cells, androgen presenting cells.
14:30
These are then presented to T cells.
14:32
T cells then infiltrate into the tumour and show basically increased levels of infiltration after treatment.
14:39
And this is a way of basically from them, they've got an intratumoral treatment, but they see a systemic effect.
14:47
So we think we'll see, you know, this type of effect will be beneficial that it gives you then a sustained response.
14:52
So you know, once you've got over the initial treatment with the nanoparticles, the immune system is basically then activated for a longer term response.
14:59
So that's kind of our angle that we're looking at for this more sustained response, but also the ability to target tumours that are more deep-seated.
15:06
So pancreatic talked about triple negative breast, prostate areas that are traditionally difficult to treat.
15:14
That's it.
15:15
Thank you.