EP-4734968-A1 - CALIXARENE-BASED DELIVERY SYSTEM AND METHOD OF USE

Abstract

The current invention relates to a delivery system to deliver one or more cargo to one or more cells, wherein the cargo delivery system comprises at least a calixarene, a phospholipid, an additional lipid such as sterol. The invention further relates to a method of delivering cargo to a subject using the delivery system and a pharmaceutical composition comprising the delivery system. The invention also relates to the use of a calixarene in an immunogenic composition, wherein said composition comprises an immunogenic component encapsulated in a lipid nanoparticle (LNP) comprising said calixarene and wherein said LNP has an adjuvant effect in said immunogenic composition. The invention also relates to a vaccine, wherein said vaccine comprises an immunogenic component encapsulated in a lipid nanoparticle, wherein said lipid nanoparticle comprises at least one calixarene molecule and said lipid nanoparticle acts as an adjuvant in said vaccine. The invention also relates to a method of preparing an immunogenic composition and a composition comprising a lipid nanoparticle (LNP) adjuvant comprising calixarene.

Inventors

• VANDER STRAETEN, Aurélien
• BEVERNAEGIE, Robin

Assignees

• Phoenix Biosciences SA

Dates

Publication Date

20260506

Application Date

20240429

Claims (20)

1. 1. A delivery system to deliver one or more cargo to one or more cells, wherein the delivery system comprises a calixarene, a phospholipid, and an additional lipid wherein the additional lipid is selected from a sterol, a fatty acid, a glycerol monooleate, a trioleate or a short saturated molecule.
2. 2. The delivery system according to claim 1, wherein said calixarene is a calix[4]arene.
3. 3. The delivery system according to any of the previous claims, wherein said calixarene is an ionizable calixarene, a cationic calixarene or any combination thereof.
4. 4. The delivery system according to any of the previous claims, further comprising an ionizable lipid.
5. 5. The delivery system according to any of the previous claims, further comprising a PEGylated lipid and/or a polysarcosine.
6. 6. The delivery system according to any of the previous claims, wherein said system comprises a calixarene, a phospholipid, a sterol and a PEGylated lipid, wherein said calixarene is an ionizable calixarene.
7. 7. The delivery system according to claim 6, wherein said calixarene is a calix[4]arene bearing one or more ionizable head groups.
8. 8. The delivery system according to claim 7, wherein one or more of said ionizable head groups comprises ternary amine groups.
9. 9. The delivery system according to any of the previous claims 7-8, wherein said calixarene comprises 4 identical ionizable head groups.
10. 10. The delivery system according to any of the previous claims 6-9, wherein said ionizable calixarene is present in said delivery system at a mass fraction of 10-60 % (w/w).
11. 11. The delivery system according to any of the previous claims 6-10, wherein said phospholipid is present in said delivery system at a mass fraction of 5-35% (w/w), more preferably 10-30% (w/w).
12. 12. The delivery system according to any of the previous claims 6-11, wherein said sterol is present in said delivery system at a mass fraction of 15-50% (w/w).
13. 13. The delivery system according to any of the previous claims 6-12, wherein said PEGylated lipid is present in said delivery system at a mass fraction of 2-24 % (w/w), more preferably 10-24 % (w/w).
14. 14. The delivery system according to any of the previous claims 1 to 5, wherein said system comprises a calixarene, wherein said calixarene is a cationic calixarene; a phospholipid; a sterol; a PEGylated lipid; and an ionizable lipid.
15. 15. The delivery system according to claim 14, wherein said calixarene is a calix[4]arene bearing 4 cationic head groups.
16. 16. The delivery system according to claim 15, wherein one or more of said cationic head groups comprises at least one quaternary amine group.
17. 17. The delivery system according to any of the previous claims 15-16, wherein said calixarene comprises 4 identical cationic head groups.
18. 18. The delivery system according to any of the previous claims 14-17, wherein said cationic calixarene is present in said delivery system at a mass fraction of 0.1-50 % (w/w).
19. 19. The delivery system according to any of the previous claims 14-18, wherein said phospholipid is present in said delivery system at a mass fraction of 5-35 % (w/w), more preferably 10-30% (w/w).
20. 20. The delivery system according to any of the previous claims 14-19, wherein said sterol is present in said delivery system at a mass fraction of 15-50 % (w/w).

Description

CALIXARENE-BASED DELIVERY SYSTEM AND METHOD OF USE FIELD OF THE INVENTION The present invention relates to a delivery system, specifically a calixarene-based delivery system for transporting various cargos, such as nucleic acids, proteins, chemical substances, and polysaccharides, to target cells. The invention also encompasses a method of delivering cargo to a subject using the described delivery system. BACKGROUND The efficient and targeted delivery of therapeutic agents, such as nucleic acids, proteins, and chemical substances, to cells is a critical aspect of modern medicine. Various delivery systems have been developed to enhance the efficacy and safety of these agents. However, there is a continuous need for improved delivery systems that can effectively deliver diverse types of cargo to target cells. RNA therapeutics comprise a rapidly expanding category of drugs that will change the standard of care for many diseases and actualize personalized medicine. These drugs are cost effective, relatively simple to manufacture, and can target previously undruggable pathways. However, employing nucleic acids as therapeutics is challenging because they are susceptible to degradation by nucleases, contribute to immune activation and have unfavorable physicochemical characteristics that prevent facile transfection into cells. Safe and effective nucleic acid therapeutics therefore require sophisticated delivery platform technologies. Lipid nanoparticles are the leading technology for nonviral nucleic acid delivery. Naked RNA is quickly degraded after administration by cellular ribonucleases (RNases). LNPs slow down the degradation process to ensure RNA stability while also promoting cellular internalization via endocytosis and allowing intracellular release of RNA into the cytoplasm for translation by cellular machinery. The composition of LNPs typically includes an ionizable cationic lipid and three neutral helper lipids: phospholipid, cholesterol, and lipid-anchored polyethylene glycol (PEGylated lipid). The ionizable cationic lipids are complexed with polyanionic RNA through ion pairing interactions to enable its encapsulation by the neutral lipids and facilitate cellular uptake and endosomal escape. LNPs were initially optimized for formulating siRNA (~23 nt) and have recently evolved to encapsulate larger RNA-agents, including mRNA (-1000 nt). Recently, there has been increased activity for even larger RNA payloads, such as selfamplifying RNA (saRNA). saRNA is a promising alternative to mRNA as it has been shown to induce immune responses with up to 100-fold lower doses and extended protein expression in vivo compared to mRNA. However, saRNA (-10000 nt) is larger than mRNA (-1000 nt) and has more secondary structure, making it more difficult to encapsulate and deliver. The inherent chemical and structural differences between mRNA and saRNA in terms of length, stability, and charge density suggests that LNP delivery formulations for saRNA may require conditions significantly different from those developed for mRNA delivery. The ionizable lipid is considered to be the most important factor for improving encapsulation efficiency, as this is the component responsible for complexing with RNA cargo. However, in order to obtain a sufficient encapsulation efficiency for saRNA, high amounts of ionizable lipids are necessary (for instance 20 times the amount of ionizable lipid compared to the amount of saRNA). Optimizing the efficiency of cargo encapsulation will minimize waste of expensive materials (i.e. RNA), and generate a higher concentration drug formulation. Besides influencing encapsulation efficiency, cellular uptake and promotion of endosomal escape of nucleic acid cargo, the various constituents of the LNP are also important to facilitate monodisperse nanoparticle formation and improve nanoparticle stability. Polydispersity index (PDI) is a normalized value that indicates nanoparticle size range in a sample, and is a useful indicator of sample quality. In samples with high dispersity, larger particles in the distribution will tend to aggregate and sediment, which leads to diminished effective RNA concentration and inconsistent dosing. Typically, LNP formulations developed for biological application should have a PDI below 0.2, which indicates the colloid is acceptably monodisperse. Monodispersity of nanoparticle drugs is crucial to ensure the consistent behavior of the intended drug, as size influences how particles interact with the body. However, in order to obtain a monodisperse population of LNPs, especially for those having an saRNA cargo, high amounts of ionizable lipids are necessary. The present invention aims to resolve at least some of the problems and disadvantages mentioned above. SUMMARY OF THE INVENTION The present invention provides a delivery system for delivering one or more cargos to one or more cells according to claim 1. The delivery system comprises a calixarene, a phospholipid, an a