# US 11,077,205 B2 · Synthetic compound for improving efficiency of transfection > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 11,077,205 B2 - Document ID: US11077205B2 - Status: Granted - Kind: B2 - Application number: 16/088,730 - Filed: 2017-03-30 - Published: 2019-03-21 - Granted: 2021-08-03 - Earliest claimed: 2016-03-31 - Assignees: Klinikum der Universität München - Applicants: Klinikum der Universität München - Inventors: Shan Guan; Joseph Rosenecker - Web page: https://patentgrep.com/patent/US11077205B2 ## CPC - A61K 48/0041: {the non-active part being polymeric} - C07K 7/08: having 12 to 20 amino acids (gastrins C07K14/595; somatostatins C07K14/655; melanotropins C07K14/68) - C07K 19/00: Hybrid peptides {, i.e. peptides covalently bound to nucleic acids, or non-covalently bound protein-protein complexes} - C12N 15/87: Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation - C07K 2319/01: containing a localisation/targetting motif - C07K 2319/09: containing a nuclear localisation signal - C07K 2319/80: containing a DNA binding domain, e.g. Lacl or Tet-repressor - C07K 2319/85: containing an RNA binding domain - A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D) - C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00) - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) ## Abstract The invention concerns a synthetic compound for enabling transfection of eukaryotic cells by means of an amphiphilic block copolymer which synthetic compound comprises a peptide residue having at least one targeting sequence and a nucleic acid binding sequence which nucleic acid binding sequence comprises or consists of at least four consecutive amino acid residues which are positively charged at pH 7.4, wherein the targeting sequence is any sequence that causes a direction of the sequence to a position or a structure inside or on the surface of eukaryotic cells, characterized in that the synthetic compound further comprises a hydrophobic moiety covalently linked to the peptide residue, wherein the hydrophobic moiety comprises or consists of lipoic acid residue, lipoamide residue, a tetradecyl residue, a cholesteryl residue, or a further peptide residue having a sequence with more than 40% amino acid residues with hydrophobic side chains. ## Claims ### Claim 1 A synthetic compound for enabling transfection of eukaryotic cells, in combination with an amphiphilic block copolymer which synthetic compound comprises a peptide residue having at least one targeting sequence and a nucleic acid binding sequence which nucleic acid binding sequence comprises at least four consecutive amino acid residues which are positively charged at pH 7.4, wherein the peptide residue comprises SEQ ID Nos: SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ILS NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9 or SEQ ID NO: 10, wherein the synthetic compound further comprises a hydrophobic moiety covalently linked to the peptide residue, wherein the hydrophobic moiety comprises lipoic acid residue, lipoamide residue, a tetradecyl residue, a cholesteryl residue, or a further peptide residue having a sequence with more than 40% amino acid residues with hydrophobic side chains, the further peptide comprising at least four amino acid residues with hydrophobic side chains, wherein the amino acid residues with hydrophobic side chains are amino acid residues of phenylalanine, tyrosine and tryptophan and the targeting sequence comprises a nuclear localization signal (NLS) sequence, a skeletal muscle cell ligand, a myocardium ligand, and/or an epithelium ligand sequence. ### Claim 2 (depends on 1) The synthetic compound according to claim 1, wherein the epithelium ligand sequence is a human airway epithelium ligand sequence. ### Claim 3 (depends on 1) The synthetic compound according to claim 1, wherein the positively charged amino acid residues comprise histidine residue(s), arginine residue(s) and/or lysine residue(s). ### Claim 4 (depends on 1) The synthetic compound according to claim 1, wherein the hydrophobic moiety is covalently linked to the amino terminus or to the carboxy terminus or to any side chain of any amino acid residue of the peptide residue. ### Claim 5 (depends on 4) The synthetic compound according to claim 4, wherein the side chain is the side chain of lysine. ### Claim 6 (depends on 1) The synthetic compound according to claim 1, wherein the maximal length of the peptide residue, optionally together with the further peptide residue, is 45 amino acid residues. ### Claim 7 (depends on 1) A pharmaceutical composition comprising the synthetic compound according to claim 1 together with a pharmaceutically acceptable carrier. ### Claim 8 (depends on 1) The synthetic compound according to claim 1 for use in the treatment of a genetically caused disease of a human being or an animal by gene therapy, wherein the synthetic compound is used together with the amphiphilic block copolymer and a nucleic acid to be transferred into the cells of the human or an animal. ### Claim 9 (depends on 8) The synthetic compound for use in the treatment of a genetically caused disease of a human being or an animal by gene therapy or by an immunotherapy or for use in preventing an infection by vaccination according to claim 8, wherein the amphiphilic block copolymer is a poloxamer or a poloxamine. ### Claim 10 (depends on 9) The synthetic compound of claim 9, wherein the poloxamer is poloxamer 184 and/or the poloxamine is an ethylene oxide-propylene oxide copolymer with tetraether with (1,2-ethanedinitrilo)tetrakis(propanol). ### Claim 11 (depends on 8) The synthetic compound for use in the treatment of a genetically caused disease of a human being or an animal by gene therapy according to claim 8, wherein the compound is present in a nebulized form. ### Claim 12 (depends on 9) The synthetic compound for use in the treatment of a genetically caused disease of a human being or an animal by gene therapy or by an immunotherapy or for use in preventing an infection by vaccination according to claim 9, wherein the compound is present in a nebulized form. ### Claim 13 (depends on 1) Method for an in vitro transfection of eukaryotic cells in cell culture comprising the following steps: a1) Mixing a plurality of molecules of the synthetic compound according to claim 1 with a plurality of molecules of an amphiphilic block copolymers to allow the formation of binary complexes, b1) mixing the formed complexes with a plurality of molecules of a nucleic acid to be transferred into the cells to allow the formation of ternary complexes, or a2) mixing a plurality of molecules of a nucleic acid to be transferred into the cells with a plurality of molecules of an amphiphilic block copolymers to allow the formation of binary complexes, b2) mixing the formed complexes with a plurality of molecules of the synthetic compound according to claim 1 to allow the formation of ternary complexes and c) contacting the ternary complexes with the cells to be transfected. ### Claim 14 (depends on 13) Method according to claim 13, wherein the amphiphilic block copolymer is a poloxamer or a poloxamine. ### Claim 15 (depends on 9) The synthetic compound of claim 9, wherein the amphiphilic block copolymer is poloxamer 184. ### Claim 16 (depends on 1) The synthetic compound according to claim 1, wherein the further peptide residue has a sequence with more than 50% amino acid residues with hydrophobic side chains.