0:29
Thanks very much for everybody for taking the time to attend.
0:32
It's been a couple of years since I've spoken at this conference.
0:34
So glad to be back and see so many familiar faces.
0:37
But for those of you who don't know me, I'm Andrew Kennedy.
0:39
I'm part of the BD team here at CPC Scientific and we're a provider of peptides and oligos specialising in late phase manufacturing commercial supply.
0:50
Today I'll be taking you through a case study on peptide receptor radionuclide therapeutics.
0:55
And just because that doesn't trip off the tongue very well, we're going to call it PRRT.
0:58
So I'll take you through one of the case studies that we did, some of the method development and finally, just a few minutes at the end, basically take you through some of the products and services that we offer in case you're interested.
1:12
So what is PRRT?
1:13
So basically, it's a targeted peptide that basically peptides target receptors that are over expressed in in cancer cells.
1:24
And basically we're made-up of three different segments to this compound.
1:29
So we've got the cytotoxic radionuclide chelate, which chelates to a metal isotope separated by a linker and obviously the targeting peptide which is designed to target these species that are overexpressed in different cancer cells.
1:45
So PRRT has been around for quite a while, back in 1900 it was discovered but really wasn't until NETTER-1-trial in 2017 that really led to a series of approvals from the FDA.
1:57
And we've definitely noticed an uptake in the interest for these kind of products coming to clinical trials.
2:06
So this is basically a sharp snapshot of different peptide conjugates that are available on the market.
2:12
And so you can see that they go from kind of a simple structures all the way through to kind of more complex cyclic compounds.
2:20
So I don't really need to explain to this room why peptides are important and PRRT, but basically there's a number of factors that are considered.
2:27
So why they're beneficial compared to antibodies or proteins.
2:31
They're small size, positive pharmacokinetics, high affinities, basically less immunogenic, generally non-toxic with minimal side effects.
2:42
So this is a summary of peptides compared to proteins, antibodies and monoclonals, basically showing the advantages of using them compared to other vectors, including blood clearance, elimination routes, immunogenicity and more importantly the manufacturing costs.
2:59
They are quite cheap to manufacture compared to the other vectors.
3:04
So there's a few steps that you need to consider before heading into clinical trials.
3:10
First of all, target identification, obviously.
3:12
So identifying a specific receptor like you want to target peptide identification.
3:19
So synthetic peptide with a high affinity for that target.
3:23
Step 3, which we are heavily involved in, this is the design of the chelate essentially.
3:28
So being able to design the chelate so that it binds effectively to the receptor that has various kinds of functional groups to increase that binding and that it fits in the binding pocket essentially.
3:41
And there's a few others like selection of radionuclide, there's different ones to select the radio labelling procedure because basically when you radio label these chelates, not all of them are 100% labelled with the isotope.
3:55
So we're kind of looking for high labelling efficiency, which increases the activity and obviously in vitro characterisation in vivo assessment, and obviously the dosimetry data at the end after preclinical studies and toxicological studies.
4:10
So this is a snapshot of different chelates that are basically available that we've worked on at CPC.
4:17
I'll be talking about DOTAGA later on, but there's other ones available.
4:20
I'll also mention TRAP at the end as well and the different kind of advantages of using different chelating compounds.
4:28
And these are the different types of metal isotopes.
4:31
So there's various different types of kind of radioactive isotopes you can use having different particles waves and it also shows the tissue penetration as well.
4:41
So that's something you want to consider depending on the application.
4:46
And this is the kind of list of the peptides.
4:48
So we should be kind of familiar with most of these.
4:50
So bombesin or triotide, etcetera.
4:52
And there are indications for different types of cancer that'll be expressed in the tissue.
4:59
And here's another snapshot of peptides that are synthesised by CPC Scientific, including the targeted receptor in red.
5:07
So these are the ones that we've worked on, and we've got some citations as well, if anyone's interested in referring to any of these components as well.
5:16
So we're going to the manufacturing case study.
5:17
So this is where the DOTAGA labelled urea based PSMA inhibitor.
5:23
I'll take you through the kind of synthesis steps that we used to get to the quite high crude purity and then the kind of purification stage to get to a really high and numerically pure compound as well.
5:36
So PSMA targeted peptide therapeutics are basically heavily over expressed.
5:43
So this is why it's a good advantage to target this type of type of receptor.
5:51
So it's over expressed by about 100- to 1000-fold in normal tissue and it's expressed at all stages of the disease.
5:58
So it can be detected quite easily.
6:01
And yeah, so the transmembrane conformational structure of PSMA results in internalisation of the therapeutic and I'll go through a diagram of how that is represented later.
6:11
So these are basically types of peptide-based inhibitors that you can synthesise; phosphorus, thiol and urea.
6:22
Phosphorus and thiol have some limitations, including the ability to penetrate the blood brain barrier and insufficient metabolic stability.
6:30
So urea-based ligands are usually highly favoured.
6:34
And that's the kind of ligand I'll go through for this example.
6:39
So for the urea-based ligands, there's a couple of structural elements that you really want to consider including charge, chemical components and hydrophobicity.
6:49
And this is just to increase that lock and key effect the binding to the receptor on the linkage composition.
6:56
So we'd like to incorporate amino acids into the linkage also include multiple aromatic rings because that helps the binding to this S1 accessory pocket, which I'll show you in the diagrams later.
7:07
And also using D amino acids for enhanced stability.
7:10
So to prevent any enzymatic degradation or physiological degradation in the body.
7:15
So this is kind of a schematic of the PSMA active binding site.
7:20
There's a 20-angstrom entrance funnel and quite a long dip into the actual binding pocket.
7:25
So we've got a glutamate receptor, our glutamate functional group at the at the end of the peptide that binds to the very end of the pocket.
7:34
We've also got those aromatic rings providing increased binding to the receptor and also the length of the spacer as well has to fit long enough to be able to allow that glutamate to bind to the receptor.
7:51
So these are kind of the main kind of sections of the PSMA receptor that we synthesised, including the urea binding moiety with the glutamate motif at the end, the linker that's long enough to access the pocket and then also those aromatic rings to help bind to the pocket as well.
8:10
And obviously the most important part DOTAGA for the collating to the radionuclide.
8:17
So we went through three different methods to try and synthesise this.
8:21
So a number of stages of process development.
8:24
First of all, this is the fragment condensation kind of hybrid approach where we're coupling quartile resin, D-lysine and then on D-PHE or D-phenol, D-tyrosine and then finally with the DOTAGA coupling at the end and followed by cleavage and deprotection.
8:39
But the issue here was the actual liquid phase coupling at the end for the DOTAGA.
8:46
So it requires two cleavage steps instead of one and one liquid phase reaction.
8:52
The synthesis of the special material succinimide is relatively difficult and the purities were pretty poor.
8:59
So less than let's say 47% for yield and 49% for purity.
9:05
So method 2, this was a solid phase kind of resin bound roots that we use the lysine with DD protection.
9:14
So coupling the resin, coupling the amino acids going right through to the coupling of the DLys(Dde) and then the target at the end.
9:24
And this yeah.
9:26
So the disadvantages were the synthesis of our special material is relatively difficult.
9:31
So the purity low to begin with. This ended up with a yield of 66 and purity of 51.
9:37
So still not great and similar to the first method.
9:41
But finally we ended up with the solid phase preparation of the resin bound protected peptides and so on resin.
9:48
We then use these fragments and so the Lys-ureido fragment followed by D-lysine and D-Phe-D-tyre and then followed finally by the DOTAGA coupling at the end.
9:59
So this was quite successful.
10:01
So for a number of reasons, the synthesis route was successful to begin with.
10:05
But we also have some proprietary cleavage technology that allows for fast and efficient cleavage of the peptide and of the protecting groups and also the total yield.
10:16
We've also got proprietary purification technology which not only purifies but also non-numerically purifies the material as well.
10:23
So the, the actual crude purity was about 85/90% and the final purified product was close to 100.
10:31
So this is a summary of the three methods.
10:33
So fragment condensation, the hybrid approach, the Dde protection and the final tBu protection.
10:41
So obviously number 3 was the winner of this process development.
10:46
And these are examples of the chromatogram.
10:48
So really, I mean, crude couldn't ask for better than that really.
10:51
And then purified and again, numerically pure as well.
10:54
I haven't shown the EE here, but it's 99.3% for purity with pretty much close to 100 and non numerically pure as well.
11:02
So to give you an idea of how long a typical project like this might take so we can get to for that particular compound to synthesise, you know, multi gramme batches take around 10 months, which is actually quite a quick timeline for this type of project.
11:18
And it's really a benefit of carrying out the on a local method development and the peptide process development in parallel and then followed by the multi gramme production.
11:28
I'm using proprietary cleavage technology and the purification techniques gives high yield, high purity and then managed to do the IND submission within 10 months.
11:38
So there's a number of citations here for other peptide conjugate projects.
11:44
If you're interested in taking them, please stop by the booth, then we can chat about the different references.
11:49
There's also a number of different citations that CPC has.
11:52
Obviously won't go through all these, but we've been working on this for quite some time.
11:56
And these are basically researchers and groups that have used our products for this type of application.
12:01
I just want to touch on this as well.
12:04
So the TRAP is another type of collating ligand.
12:08
And we've been working on a couple of compounds here.
12:11
But just to mention, if you are working in this area, TRAP is more stable than DOTAGA over a large pH range, chemically inert in acidic and alkaline conditions.
12:21
It's got high selectivity for Ga(3+)
12:25
And that's really important for when you're actually labelling the isotope to give a high labelling selectivity.
12:33
And also this complex is done quickly within minutes and quantitatively even at a really low pH as well.
12:40
So that kind of goes through the case study.
12:41
I'll give a quick introduction to CPC because I think not many people are too familiar with our products.
12:48
So my colleague Tim here, he's here as well.
12:51
He's Senior VP of Strategic Portfolio and I'm part of the BD team along with a number of others.
12:57
Diego covers Europe as well and the others are based in the US. To give you an idea of our kind of history.
13:06
So CBC was founded back in 2001 by Sean Lee, who also founded American Peptide, if you're familiar with them.
13:12
And this kind of gives us a timeline of the last 20 years of the expansion of our Hangzhou site in China.
13:19
And also just showing the number of FDA inspections that we've had over the previous years, probably had about 7 in the past 15-20 years.
13:28
And we're also inspected by a number of other regulatory agencies throughout the globe.
13:33
We also have our own drug master files for a number of generics that we sell commercially throughout the world as well.
13:41
And recently we've also broken ground on a new manufacturing facility in the US and that should be open in 2025.
13:49
So, and also the expansion of the Hangzhou site over those years as well.
13:54
So in all of our manufacturing at the moment, it's carried out in Hangzhou in China.
13:57
Our offices are based in San Jose, CA.
14:00
And then as I say, we've broken ground on the site in Rockland.
14:03
Well, that should be up and running for peptides phase I and II early next year.
14:07
So giving an overview of the campus for the Hangzhou site.
14:11
So this is quite a large area, 700,000 square feet of production with close to 500 scientists now, isolated process suites for synthesis, cleavage purification as you'd expect and capacity of about half a tonne per year.
14:24
And we also started our oligo synthesis platform about 3 or 4 years ago.
14:29
And that really was because we got a lot of interest in peptide oligo conjugates and we decided to bring the oligo platform in house.
14:36
So we've now got quite an experienced group to do oligos for various different applications as well.
14:42
So we can do that up to GMP, up to just under a kilogramme scale.
14:46
So that's kind of a picture of the Rockland facility, our site, our offices in San Jose.
14:53
And so thank you very much for your attention.
