Combinations of mrnas encoding immune modulating polypeptides and uses thereof
A technology of encoding and composition, applied in immunotherapy methods and compositions, polynucleotides of interleukin-23 polypeptide, encoding interleukin-36γ field, capable of solving problems such as undesired immune response and safety
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[1177] Formulations of the pharmaceutical compositions described herein may be prepared by any method known or hereafter developed in the art of pharmacology. Generally, such preparation methods include the steps of bringing the active ingredient into association with excipients and / or one or more other auxiliary ingredients, and then, if necessary and / or desired, dividing, shaping and / or packaging the product as Single-dose or multiple-dose units as desired.
[1178] The relative amounts of the active ingredients, pharmaceutically acceptable excipients and / or any additional ingredients in the pharmaceutical composition according to the present disclosure will depend on the identity, size and / or condition of the subject to be treated and further depend on The route of administration of the composition will vary. By way of example, the composition may comprise between 0.1% and 100%, such as between 0.5% and 50%, between 1% and 30%, between 5% and 80%, or at least 80% (w / w) act...
Embodiment 1
[2422] Synthesis of compounds according to formula I
[2423] A. General considerations
[2424] All solvents and reagents used were obtained commercially and used as received unless otherwise stated. CDCl at 300K using a Bruker Ultrashield 300MHz instrument 3 record in 1 H NMR spectrum. for 1 H, Chemical shifts are reported as parts per million (ppm) relative to TMS (0.00). Silica gel chromatography was performed on an ISCO CombiFlash Rf+Lumen instrument using ISCO RediSep Rf Gold Flash Cartridges (particle size: 20-40 microns). Reverse phase chromatography was performed on an ISCO CombiFlash Rf+Lumen instrument using a RediSep Rf Gold C18 high-efficiency column. By using a Waters Acquity UPLC instrument with DAD and ELSD and a ZORBAX Rapid Resolution High Definition (RRHD) SB-C18LC column (2.1 mm, 50 mm 1.8 μm) and a gradient of 65% to 100% acetonitrile in water containing 0.1% TFA All final compounds were determined to be greater than 85% pure by reverse phase UPLC-MS...
Embodiment 2
[2494] Generation of Nanoparticle Compositions
[2495] A. Generation of Nanoparticle Compositions
[2496] Nanoparticles can be mixed using mixing methods such as microfluidics and T-junction of two fluid streams, where one fluid stream contains the polynucleotide and the other fluid stream has the lipid component.
[2497] The lipid composition is prepared by combining in ethanol at a concentration of about 50 mM: an ionizable amino lipid disclosed herein, for example a lipid according to formula (I); a phospholipid (such as DOPE or DSPC, available from Avanti Polar Lipids, Alabaster, AL); PEG lipids (such as 1,2-dimyristoyl-sn-glycerol methoxypolyethylene glycol (also known as PEG-DMG), available from Avanti Polar Lipids, Alabaster, AL) and structural lipids (such as cholesterol, available from Sigma-Aldrich, Taufkirchen, Germany, or corticosteroids (such as prednisolone, dexamethasone, prednisone, and hydrocortisone) or combinations thereof). Solutions should be refriger...
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