0:00
We have Iolanda Micco, head of discovery chemistry from Axxam, and she's going to talk to us about targeting P2X7 from early discovery to a clinical stage candidate. Very exciting.
0:17
Thank you.
0:24
Thank you for the introduction.
0:26
OK, so let's start from a few words: Axxam is a science driven drug discovery company.
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We are in Italy.
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We were founded in 2001, and our headquarters is in Milan, and we also have additional chemistry labs in Naples, and business offices in Cambridge in the US, Copenhagen, and also here in Basel.
1:01
So we work in the drug discovery process and specifically in the early segment of the drug discovery process covering from the target identification on to the lead or the preclinical candidate selection.
1:16
So during the years we have developed a series of platforms and skills in-house.
1:25
We are expert in vitro biology, generation, validation, optimization of vitro assays which can be a cell based assays and cell free assays.
1:38
We as everyone work constantly the evolution of science which can be chemistry or the biology.
1:47
So we have introduced very niche biology platforms such as, you know, some optogenetics, iPSC cells, organellar electrophysiology, RNA biology.
2:00
We put also everything together to offer a pretty comprehensive package of for hit identification because the drug discovery process of course is a complicated process, and it goes through different phases.
2:21
But no matter what the hit identification phase is the first phase where we actually select the chemical matter that we bring forward. A mistake out there that's paid pretty heavily later.
2:35
So it's important to pay attention to what we do, what we select and how we select the chemical matter at that stage.
2:44
So we put a lot of effort trying to really boost that phase and putting all the disciplines that are required to select the best hit.
3:02
By doing that we also included different tools which for example in the last year we introduced the virtual screening and the hit that come from whatever screen that invalidated and qualified in house also from the chemical and the preliminary ADME perspective.
3:22
So we basically want to have all the needs to select inform the decisions our hits.
3:31
We also support our client as they hit to lead face and ultimately we have our compound libraries, compound management facilities which take care of our internal libraries, but also our clients’ libraries.
3:48
So during the years we put everything together trying to bring values into the scientific community.
3:57
So Axxam, many years ago had its own proprietary research pipeline and during these years, we have sort of generated new assets that were then transferred into Golgi Neurosciences.
4:16
So back in 2023, Golgi Neurosciences kind of split from Axxam and then they transferred those assets either in exchange of equity with existing companies such as Acousia Therapeutics, Muna Therapeutics or Breye Therapeutics, or they just used those assets to generate new companies.
4:40
And that was the case for example, for Lario, for Rewind Therapeutics or for Libra Therapeutics.
4:46
So the story I'm going to talk about today is the story of three main players, Axxam, Golgi Neurosciences and Breye. It’s one of those examples of molecules usually from bench to clinic.
5:03
It's one of my favourite.
5:06
So why Breye Therapeutics?
5:09
Breye Therapeutics which is a Danish company is really focused on the development of new therapies for vascular retinal diseases such as diabetic retinopathy or age-related macular degeneration.
5:29
And why is that? Because of the current treatment for those diseases are really first of all are limited and then second of all, they have really bad compliance for the client because most often they require intravitreal injections.
5:49
So the goal here for that company is to identify oral treatment, which would be amazing if you can have an ocular exposure, high ocular exposure for a company. That it doesn't need an injection.
6:07
So why P2X7? P2X7 is a purinergic receptor, is a ligand gated cation channel, it's activated by high concentration of ATP.
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Now the important thing of this receptor is that is first of all its localisation is found in CNS and also in photoreceptors, and immunocompetent cells, and is involved in inflammation cascades.
6:46
So the activation of P2X7 due to high concentration of ATP will eventually, ultimately bring an excess of proinflammatory cytokines or whatever other signalling molecules.
7:05
So for example, if we see in the microglia, P2X7 is found in microglia and in there it was the secretion of interleukin 1 beta.
7:20
And for that, it has also been considered potentially an interesting target for Alzheimer's disease.
7:28
And we also actually tested that during this programme as well.
7:34
But it's also found in the eye, as I was saying, and specifically in photoreceptors and Muller cells.
7:43
Now here the ATP readings that increases during hyperglycaemia that will activate the P2X7 and eventually that will cause the release of cytokines TNF alpha, interleukin 1 beta, also VEGF.
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So eventually that will bring like a worsening of the disease.
8:13
So potentially blocking the P2X7 can really be, yeah, an interesting way to go for this disease.
8:25
So the programme started years ago in Axxam.
8:29
As I was saying, we ran 2 high throughput screening in-house.
8:34
We identified some interesting hits that were brought into the hit to lead phase and lead optimization.
8:41
Everything was ran in Axxam and now as I was saying is in Breye’s hands and they are trying to develop the molecule that was born in our labs and potentially bring it into the clinics very soon.
8:59
So I'm trying to see what we did and sorry I cannot show you a lot of things, but I tried to do what I could.
9:10
So the hit identification phase, as I mentioned, we predicted the assay generation in house, and we ran this through high throughput screening campaigns which provided actually a multiple of genotypes.
9:29
As I said before, this is an important phase of the drug discovery phase.
9:35
So we put a lot of effort trying to narrow down the two chemical classes that we wanted to bring forward.
9:44
And here, I reported 2 molecules, 1, on the left hand side, representing the main class and the other one representing the backup class.
9:57
So the main class was brought into the lead generation phase where we obviously, as always tried to improve the potency.
10:05
That's what we always do.
10:09
But together with that we worked also on other physical chemical parameters and ADME profile.
10:15
We were we generated about 500 molecules which were designed and synthesised in Axxam.
10:22
I cannot show you the structure of the new compound, but as you can see we sort of worked in the ethereal cycle on the left hand side of the molecule.
10:32
They preparedly infused with the ring in the centre and also the substitutions and potentially also other heterocycles in that position in the right hand side of the molecule as well.
10:46
So we identified a good lead which was then brought into the lead optimization phase where we synthesise about 100 molecules and here I had to narrow down even more of the structure.
11:01
But we identified a particular candidate which is now having further exploration.
11:11
So the way how we explore that during the hit to lead phase and also the following phase is through a classical iterative cycle.
11:28
We investigated the potency of the molecules, trying to identify the most interesting molecules, ran FLIPR assays, but also [unclear] assays with QPatch and also secondary assays, evaluating also the impact in vitro on as I said before cytokines and other signalling molecules involved in the inflammation path.
11:57
We investigated the selectivity towards P2X and P2Y, which was not a problem for that class to be honest, never an issue also for hERG.
12:06
Whereas we found some difficulties for the DDI part of it, because there was no CYP inhibition problem, but some of the molecule had some induction.
12:18
So we had to work a lot trying to figure out the way how to abolish that.
12:25
And obviously we managed not a problem on permeability. Whereas on the metabolic stability, some of the molecules were showing some tendency to be pretty [unable].
12:41
So overall, the lead compound really showed a pretty robust profile with a pretty very high potency in the in vitro assays, clear potency also in the secondary assays, no ADME problems, no hERG liabilities.
13:03
And as I said before, where some of the molecules had some of the CYP induction, the molecule was completely clean and interestingly the PK, which was, you know, as I said before, all those things without a good PK, oral PK, wouldn't have been enough at all.
13:24
We're really happy to see that the PK of either mouse or rat were actually pretty good also after oral administration.
13:34
So that gave us the confidence to explore.
13:37
I didn't put the data on the PK, but I decided to show you that the exposure in the eye, which in the end is what counted the most.
13:47
So the good PK in rat and mouse brought us to explore even more.
13:52
And we did this exploration on the exposure, the ocular exposure in rats.
13:58
As you can see, we explored, we investigated the concentration on retina, optic nerve, and pigmented epithelium.
14:08
And in all cases we had an excellent exposure.
14:11
The concentration of the molecule was way or a few folds higher than the effective IC50 in vitro.
14:20
So we were, the team was really happy with that.
14:29
We studied also the in vivo pharmacology. I reported here this maybe the modulation of the interleukin beta and the TNF alpha after 50 and 100 milligrammes per kilo where you can see that both interleukin beta and TNF alpha, they basically decrease after four and eight weeks of treatment.
14:59
On the bottom side of the slide, you can also see that the glucose level were also checked.
15:09
First of all because we wanted to make sure that the glucose levels were up.
15:15
So the condition of diabetes was actually up there, but you also want to check that the molecule doesn't have an impact on the blood glucose, because you want to make sure that what you are seeing is not because glucose is coming down and the ATP is going down.
15:35
You want to make sure what you're seeing in your phenotypic results are actually due to the activity of the receptor.
15:53
So the main serious progress from compound one which was the hit compound into a preclinical candidate was really exemplary, it was started from a high nanomolar potency, but it was still a three digit nanomolar [unclear].
16:22
The poor solubility and with a few liabilities which were then solved at least partially going, you know from the hit to the lead.
16:32
So from compound 1 to compound 2 where the potency was improved, the ADME the was also improved and we had this excellent exposure in our ultimately the preclinical candidate compound number 3 showed a pretty good potency, actually was at the same level of compound 2.
16:55
But there was no, and there are no issues whatsoever.
16:59
So we are really confident that the molecule can continue its path. On the backup class, as I said there was a backup class that was also brought forward.
17:17
Compound 4 was the hit that was identified during the high throughput screening campaign with low micromolar potency, still a good IP, lower solubility and stability, that was further improved.
17:35
As you can see the benzol portion was replaced in the compound 5 to obtain validated hit with improved advancing and chemical properties obviously.
17:48
And then compound 6 which represents the most advanced molecule in the series is really shows still a good potency.
18:00
Now we are in the nanomolar range as well and the promising ADME as well.
18:05
Still we had some liabilities in terms of CYP induction.
18:10
So that's something that is kind of slowing down this backup series.
18:18
So during this programme we have generated about five patents where we covered the most important, the most promising molecules either identified after the high throughput screening or generated later.
18:36
And the last one has been deposited for the preclinical candidate molecule.
18:47
So I think in conclusion, this is an example of programme from hit identification to the preclinical candidate selection.
18:58
It really showcases Axxam’s jet engine for these integrated programmes. We identified 2 main classes.
19:13
One of them made the run very quickly into the preclinical phases.
19:20
And sometimes when we say here, when we tell stories, everything looks pretty easy and straightforward.
19:31
We all know that it's never the case.
19:34
It was complicated.
19:35
It required efforts, resilience, patience, maybe 5 platforms, 7 disciplines.
19:41
But we say that it was easy.
19:44
But I think if there is one thing I've learned in my life is that great science starts with great ideas, but great ideas always come from happy teams.
19:57
So when we tend to tell our people to read papers and increase their knowledge and their skills, let's not forget to let the team have fun together.
20:10
I think it's the first element.
20:12
So that's what we do all the time and while we work on programmes and have fun and work very hard.
20:20
We never forget to go out, have fun together and.
20:26
Know each other also on other from other angles.
20:31
So I think that's probably all.
20:35
I'm going to give a special thanks to all the people in Axxam that contributed to the success of this programme.
20:43
Also, all the others that did not contribute to this programme but worked really hard every day.
20:49
I want to thank Breye Therapeutics’ team that allowed us to use the story, which is not always the case.
20:57
So I'm really grateful.
20:59
I would like to thank Golgi Neuroscience not only for the great collaboration but also to be an example of company creator.
21:10
I also want to thank the other CROs that contributed into part of this programme like Symeres and WuXi, and of course all of you for your attention despite the fact that it's lunch time.
21:27
So thank you.
