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So welcome back.
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I would like to invite Soehnke Voss to for his presentation.
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Soehnke holds a PhD in biochemistry from the Eberhardt Corals University in Tubingen, where he specialised in immunology and organic chemistry.
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His doctoral research conducted under Professor Gunter Jung focused on the development of toll like receptor 2 and 4 modulators for use in septic shock treatment and vaccine adjuvants for the solid foundation R&D.
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Soehnke has held roles at EMC Micro Collections, CureVac and Evonik Industries. Since joining Croda in 2007, he has progressed from account manager to his current role of business development in EMEA region for Croda Pharma, where he leverages the scientific industry expertise to drive strategic growth.
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Thank you.
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Thanks so much.
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Yeah, thanks for the nice introduction.
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I’m business development manager as at Avanti and Croda.
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So I would speak about lipids and what we are doing at Avanti and yeah, starting with this slide actually.
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So lipids have raised a lot of attention in the past years due to its ability to deliver RNA.
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But this molecule, these molecules are much more versatile.
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And so they provide delivery solutions for all kinds of APIs like proteins, adjuvant systems, small molecules and so on.
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And using different types of cargos like small vesicles for instance, or the LNPs and multi laminar vesicles for sustained release and things like that.
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So quite interesting molecules.
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And today I would like to focus on LNPs.
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So lipid nanoparticles they are distinctively different to other lipid based delivery systems because of its electron dense core and composed of cationic lipids bound to and negatively charged nucleic acids surrounded by structural components and typically use for lipids to generate an LNP with different functional properties.
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So sterols and structural lipids are providing structure, shape and stability.
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The cationic lipids sequester the nucleic assets and provides facilitate facilitates and the zonal release.
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The PEG lipids avoids or prevents the application of the system and the clearance from circulation.
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So as state-of-the-art advances, we see an increased need for delivery of nucleic acid at Avanti, with increased or new challenges in the field.
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And I put some of these challenges on the slides.
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So this can be, for instance, stability at higher temperatures.
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You know that the LNPs performing our team’s needs to be cooled down for storage temperatures, for instance varying payloads.
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You can formulate siRNA, very small molecules in comparison to mRNA, but also quite large RNA molecules like saRNA.
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For instance.
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The PEG dilemma, some customers want one PEG, others don't want one PEG molecules because of some drawbacks.
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I come to this later and targeted delivery is a big topic at the moment.
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And also this will be addressed in a few slides and other routes of administration, for instance, LNPs for inhalation.
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So these are the topics.
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So Avanti is providing and developing lipids for all kind of these challenges.
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And I would like to address these challenges, and I would like to go through some of our novel lipids or solutions that we can offer.
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I'm starting with the structural lipids.
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So DSPC is the most commonly used lipid in LNP formulation in all commercial drugs at the moment.
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So it makes very stable bilayers.
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So it has a cylinder shape actually due to its saturated fatty acids acid chain and a quite large head group.
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And if you include double bonds into the lipids and reduce the size of the head group, then you get inverted hexagonal structures actually.
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So this promotes inverted hexagonal structures that is connected with higher transfection efficacy.
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So changing the phospholipid already changed your or is able to change your system.
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Coming to the cationic lipids, this has an impact on optimization of the system for specific cargoes.
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So we differentiate three types of cationic lipids.
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A fixed cation cationic has a positive charge of a wide range of pH, proven to be very good transfection reagents.
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One example, DOTAP for instance, often used with DNA delivery as a cargo.
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You see some toxic effects at higher concentrations and so we need to consider this in your formulations.
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Ionizable lipids are less toxic than fixed cationic lipids.
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The PKA is between 6 and 7, which is mandatory to have a neutral charge in circulation and a positive charge in the endosome to enable endosomal release.
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There are also multivalent and cationic lipid.
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One example from our portfolio is MVL5 and having 5 positive charges.
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It has been shown to be beneficial in combination with siRNA for instance.
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Some novel lipids from our portfolio are shown here.
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So EPC.
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Ethylphosphatidylcholine is the first cationic lipid that is derived from a natural component and it's ethylphosphatidylcholine.
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So it's a tri-ester ethylphosphatidylcholine shown to be biodegradable in the system and have a low toxicity.
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Another one is DC cholesterols or cationic derivative of cholesterol shown to be very good transactional reagent in combination with DOPE.
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So that's some examples coming to the PEG lipids and alternatives. Historically, PEG is the yeah, most prominent lipids polymer conjugate prevents proteins from binding to the LNPs and aggregation.
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However, there are some drawbacks.
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So as mentioned some customers want to avoid or these or formulators want to avoid these and because of the in induction of anti-PEG antibodies after repeated administration.
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So this may happen also it interferes with transfection efficacy.
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So 1 alternative that came up in the past years is lipids with the polymer of sarcosine.
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So amino acid, non-biogenic amino acid that demonstrate similar stealth properties like the PEG lipid, but improved transfection, a better safety profile.
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So I have put some of our poly-sarcosine lipids in the slide.
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Cholesterol is a major component in the LNP as you know 30 to 40% and in all commercial drugs.
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But a lot of research has been done to find alternatives to cholesterol and one of the most promising candidates is beta sitosterol, also from our portfolio substitute or a beta sitosterol substituted LNPs and have comparable size and encapsulation efficacy, but also show higher transfection efficacy and makes them a very interesting component if you look for alternatives here.
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It has also been successfully used for LNPs for nebulization for an inhaled administration.
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So there's the paper on that yeah, really shows this nice application for this material.
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Coming now to targeting delivery, very hot topic.
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We get a lot of requests on this topic actually.
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So the liver is the main organ for accumulation of LNPs and after intravenous administration, but also observed with subcutaneous or intramuscular administration.
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This can be beneficial for some drugs.
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So if you want to use the liver as a production factory for your protein, other drugs would benefit from extrahepatic delivery to specific organs or cell types.
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And there are two main strategies for targeting delivery.
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One is active targeting, meaning you bind the targeting molecule to the outer surface of the particle or passive targeting to change the physical chemical properties of the LNP to make it suitable for organ trafficking. And so for active targeting, head group modified PEG lipids are commonly used here either with functional groups that are able to bind to functional groups in the targeting molecule.
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Another option is to use lipid that are suitable for click chemistry.
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If you look for by orthogonal coupling which is more specific.
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Don't under mild conditions.
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However you need to introduce the counterpart.
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So if you have a DBCO lipids then you need an outside head site targeting molecule with an outside moiety actually.
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And the last way is to use lipids that already contain a targeting ligand, which is most convenient.
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And I will show some examples on the lipids that we have in our portfolio so far.
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The classical way we recommend PEG lipids with maleimides to bind to thiol groups, for instance. Carboxylic acid making mean carboxylic acid congregation amines and so on.
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So there's a large range of molecules for the click chemistry.
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We have lipids with DBCO, but also with outside moieties.
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And for click chemistry, as mentioned most convenient you use lipids that already contain targeting and moieties.
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I have 4 examples here.
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1 is folate, so the latest used to target the folate receptor which is over expressed on cancer cells of epithelial origin.
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So the strong affinity between folate and folate receptor leads to an increased uptake of the particle actually when you use the system and due to receptor or mediated endocytosis.
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Yeah, some examples of lipids that we have GalNAc and already discussed today or heard it several times today.
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So it binds to the asialoglycoprotein receptor over expressed on liver cells that can be used for liver targeting.
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The binding affinity increases when you use a multivalent ligand.
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And actually we have two molecules in our portfolio that with a triGalNAc conjugation, tri-mannose.
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So the mannose receptor is a potential target for cancer cell delivery.
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So also here we have tri-mannose lipids.
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It was also successfully used to deliver antigens or mRNA coding for antigens to APCs.
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Because APCs, specifically dendritic cells, have increased expression of mannose receptor and makes them suitable molecule.
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Lipopeptides, so we recently launched to lipopeptides containing the RGD and tripeptide, which is used from by several viruses to attach and for attachment and uptake into cells.
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And this actually lipids or RGD lipids can also be used to, for an increased cellular adhesion and uptake.
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Coming now to passive targeting.
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And so mainly the ionizable lipids plays an important role here.
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And we are very proud to start a collaboration with a company called Certest.
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We developed ionizable lipids suitable for passive targeting.
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And so we have now three Certest lipids in our portfolio and one for lung tissue, one for liver, and one for spleen targeting, passive targeting.
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So taking everything together.
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So this was yeah, a small number of lipids that we have in our portfolio, but we have much more.
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So over 2000 lipids in research quality and we are very innovative and always generate new lipids.
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So it absolutely makes sense to look on our web page what is new we can offer, and we also do lipids in GMP quality.
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So we have over 25 lipids that we can manufacture in GMP quality and all lipids that are used for LNP formulation, liposomal formulation and also lipids with targeting moiety.
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So we would have a maleimide PEG lipid in GMP quality by end of the year.
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And if a lipid is not available in research or in GMP quality, so we can make it custom.
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So if you have a custom proprietary lipid, for instance, that you want to have in research or GMP quality, we're happy to make it.
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And we also support formulation and work with our formulation experts.
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I think it was quite fast.
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Is that correct?
17:47
So if you have any questions, happy to take them.
