RU2012158391A - MODULAR MOLECULAR CONJUGATE FOR DIRECTED DELIVERY OF GENETIC STRUCTURES AND METHOD FOR ITS PRODUCTION - Google Patents

Abstract

1. A modular molecular conjugate for targeted delivery of genetic constructs based on peptide ligands of cell receptors, including ligand and cationic components, connected through a hydrozone derivative of oxocarboxylic acid of the formula: 2. The modular molecular conjugate according to claim 1, comprising, as a ligand component, the peptide X-R-His-R-Ser-R-D-Lys-Leu-Arg-Pro-R, where R is pGlu; R-Trp, D-Trp, Pro, or D-Pal (3); R is Tyr, or Arg; R-Gly-NH, D-Ala-NH, or NH-CH-CH; X is a hydrogen atom, or lower alkyl group (C-C), the cationic peptide H-Progo-Lys-Lys-Lys-Arg-Lys-Val is attached to the e-amino group D-Lys at position 6 of the peptide via a hydrazone derivative of pyruvic acid. 3. The modular molecular conjugate according to claim 1 or 2, comprising the Arg-Gly-Asp-Phe-OH peptide as the ligand component, the cationic peptide H-Pro-Lys-Lys-Lys-Arg-Lys-Val is attached to the N-terminal amino group of the peptide via a hydrazone derivative of pyruvic acid. 4. The method for producing the modular molecular conjugate according to claim 1, which consists in synthesizing a peptide ligand component and a peptide cationic component using solid-phase peptide synthesis on a Wang polymer using the Fmoc / But strategy, add oxocarboxylic acid to the peptide ligand, using pyruvic or levulenic acid then, in an aqueous solution of trifluoroacetic acid, the keto peptide is separated from the polymer carrier, and the peptide cationic stavlyayuschey 2-chlorotrityl resin with a content of C1 1.55 mmol / g using Fmoc / But-strategy, then the peptide is modified cationic component in a reaction mixture comprising a solvent D

Claims (4)

 1. A modular molecular conjugate for targeted delivery of genetic constructs based on peptide ligands of cell receptors, including ligand and cationic components, connected through a hydrozone derivative of oxocarboxylic acid of the formula:

2. The modular molecular conjugate according to claim 1, comprising as a ligand component the peptide XR ¹ -His-R ³ -Ser-R ⁵ -D-Lys-Leu-Arg-Pro-R ¹⁰ , where R ¹ is pGlu; R ³ is Trp, D-Trp, Pro, or D-Pal (3); R ⁵ is Tyr or Arg; R ¹⁰ is Gly-NH ₂ , D-Ala-NH ₂ , or NH-CH ₂ -CH ₃ ; X is a hydrogen atom, or a lower alkyl group (C ₂ -C ₅ ), the cationic peptide H-Progo-Lys-Lys-Lys-Arg-Lys-Val is attached to the e-amino group D-Lys at position 6 of the peptide via the hydrazone derivative of pyruvic acid acids. 3. The modular molecular conjugate according to claim 1 or 2, comprising the Arg-Gly-Asp-Phe-OH peptide as the ligand component, the cationic peptide H-Pro-Lys-Lys-Lys-Arg-Lys is attached to the N-terminal amino group of the peptide -Val via a hydrazone derivative of pyruvic acid. 4. The method for producing the modular molecular conjugate according to claim 1, which consists in the synthesis of a peptide ligand component and a peptide cationic component using solid-phase peptide synthesis on a Wang polymer using the Fmoc / But strategy, oxocarboxylic acid is added to the peptide ligand, in which pyruvic acid is taken or levulenic acid, after which, in an aqueous solution of trifluoroacetic acid, the keto peptide is separated from the polymer carrier, and the peptide cationic is synthesized at the same time component on a 2-chlorotrityl polymer with a C1 content of 1.55 mmol / g using the Fmoc / But strategy, after which the peptide cationic component is modified in the reaction mixture containing DMF, DIEA in an amount of 2 equivalents, Boc-NH-NH ₂ in an amount 2 eq. and HCTU in an amount of 1.1 equiv., for 3 hours with continuous stirring, after which evaporation is carried out in a rotary evaporator, the resulting residue is triturated with water, filtered and dried in a desiccator over P ₂ O ₅ pairs, after which the protective groups of the keto peptide are exposed a mixture containing a solvent of TFA, TIS and H ₂ O in a ratio of 95: 2.5: 2.5 for at least 3 hours until they are completely removed, after which the solvent is evaporated on a rotary evaporator, and the remaining oily residue of the peptide cationic component is poured with diethyl ether, precipitated k is filtered, washed with diethyl ether and dried in a vacuum desiccator, then a conjugation reaction of the obtained ligand keto peptide and hydrazide derivative of the cationic component in an aqueous solution is carried out using a five-fold excess of N-hydroxysuccinimide ether, and the formation of the corresponding hydrazones is monitored using a reversed-phase highly effective liquid chromatography.