CN-122005835-A - CRGDyk-mitochondrion composite delivery system targeting hepatic stellate cells and preparation method and application thereof

Abstract

The invention discloses a cRGDyk-mitochondrion composite delivery system targeting hepatic stellate cells, and a preparation method and application thereof. The compound system takes cRGDyk targeting peptide and exogenous mitochondria as main functional units, is covalently connected through a sulfhydryl-maleimide click chemical reaction, and introduces cerium oxide nano particles with simulated enzyme activity. The system realizes the efficient targeting of activated hepatic stellate cells through cRGDyk peptide, utilizes the exogenous mitochondrial repair cell metabolism function, and the cerium oxide nano particles provide auxiliary ROS scavenging capability, and the synergistic effect of the three can obviously reverse hepatic fibrosis in vitro cells and in vivo animal models, so that the system has good biocompatibility and wide clinical application prospect.

Inventors

• WANG FENG
• FENG WEI
• QI JING
• ZONG WEI
• LI DAI
• QIN XINYUE

Assignees

• 无锡市人民医院

Dates

Publication Date

20260512

Application Date

20260224

Claims (6)

1. 1. A cRGDyk-mitochondrion composite delivery system for targeting hepatic stellate cells is characterized by comprising a cRGDyk targeting peptide modified by sulfhydrylation, exogenous mitochondria modified by sulfhydrylation, and cerium oxide nanoparticles modified by maleimide as a connecting unit, wherein the cRGDyk targeting peptide and the exogenous mitochondria are respectively connected with maleimide groups on the surfaces of the cerium oxide nanoparticles through the sulfhydryl groups on the surfaces of the targeting peptide and the exogenous mitochondria through covalent bonds.
2. 2. The hepatic stellate cell-targeted cRGDyk-mitochondrial composite delivery system of claim 1, wherein the molar ratio of the cRGDyk targeting peptide to the cerium oxide nanoparticles is 10:1 to 50:1, and the mass ratio of exogenous mitochondria to cerium oxide nanoparticles is 5:1 to 20:1, the ratio being in a range that satisfies simultaneous synergistic performance of targeting, antioxidant activity and metabolic repair functions.
3. 3. The hepatic stellate cell targeted cRGDyk-mitochondrial complex delivery system of claim 1, wherein the complex has a hydration kinetics diameter of 600nm-900nm that avoids rapid clearance by the reticuloendothelial system while ensuring targeted delivery efficiency.
4. 4. A method for preparing the cRGDyk-mitochondria composite delivery system targeting hepatic stellate cells according to any one of claims 1 to 3, comprising the steps of S1 preparing a thiolated cRGDyk targeting peptide, S2 preparing a cerium oxide nanoparticle with a maleimide group modified on the surface, S3 preparing thiolated exogenous mitochondria, S4 incubating the product obtained in step S1 with the product obtained in step S2 to obtain cRGDyk/Ce complex, and S5 incubating the complex obtained in step S4 with the product obtained in step S3 to obtain cRGDyk-mitochondria composite delivery system.
5. 5. Use of a hepatic stellate cell-targeted cRGDyk-mitochondrial complex delivery system of any of claims 1 to 4 for the manufacture of a medicament for the prevention and/or treatment of liver fibrosis, which can target and reverse hepatic stellate cell activation, reduce extracellular matrix deposition and repair impaired cellular metabolic function.
6. 6. The use according to claim 5, wherein the medicament is administered by intravenous injection.

Description

CRGDyk-mitochondrion composite delivery system targeting hepatic stellate cells and preparation method and application thereof Technical Field The invention belongs to the field of biomedical technology and nano-drugs, and particularly relates to a novel targeted delivery system for treating liver fibrosis diseases, in particular to a composite delivery system modified by cRGDyk targeted peptide and taking exogenesis as a treatment core, wherein cerium oxide nano-particles are auxiliary antioxidant units. Background Liver fibrosis is a repair response following chronic liver injury characterized by activation and transformation of Hepatic Stellate Cells (HSCs) into myofibroblasts, leading to excessive deposition of extracellular matrix. Currently there is a lack of effective drugs that specifically target and reverse activated HSCs (ahcs). Conventional nano-drugs face the problems of low targeting efficiency, easy clearance by normal cells, single therapeutic mechanism and the like. The cRGDyk peptide is a cyclic arginine-glycine-aspartic acid peptide, can specifically identify and activate the high-expression integrin alpha v beta 3 on the surface of hepatic stellate cells, has excellent active targeting capability, but lacks a therapeutic function. Mitochondria serve as an energy factory for cells, and their dysfunction is closely related to the activation of hepatic stellate cells. Exogenous mitochondrial transplantation, an emerging alternative therapy for organelles, can effectively repair the energy metabolism of damaged cells, but is not sufficiently targeted and stable in vivo. Cerium oxide nano particles have antioxidant enzyme simulation activity and can effectively remove active oxygen, but are mostly used as dominant treatment units in the existing system, and the metabolic repair of a fibrosis root is difficult to realize by a single antioxidant mechanism. Therefore, the invention provides a novel composite system taking cRGDyk-mitochondria as a main body and cerium oxide as an auxiliary functional module, and aims to realize triple cooperative therapy of targeted delivery, metabolic repair and antioxidation. Disclosure of Invention Purpose(s) The invention aims to provide a cRGDyk-mitochondrion composite delivery system for targeting hepatic stellate cells, which can realize active targeting of aHSCs through cRGDyk peptide, repair cell metabolism function by utilizing exogenous mitochondria, remove active oxygen by means of cerium oxide nano particles, and efficiently reverse hepatic fibrosis progress under the synergistic effect of the three. Technical proposal The invention firstly provides a mitochondrion-nanoenzyme compound delivery system for targeting hepatic stellate cells, which is characterized by comprising thiolation modified cRGDyk targeting peptide, maleimide modified cerium oxide nanoparticles and thiolation modified exogenous mitochondria, wherein the cRGDyk targeting peptide and the exogenous mitochondria are respectively connected with maleimide groups on the surfaces of the cerium oxide nanoparticles through mercapto groups on the surfaces of the thiolation targeting peptide and the exogenous mitochondria through covalent bonds. Preferably, the molar ratio of cRGDyk targeting peptide to cerium oxide nanoparticle is 10:1 to 50:1. Preferably, the mass ratio of exogenous mitochondria to cerium oxide nanoparticles is 5:1 to 20:1. Preferably, the kinetic diameter of hydration of the cerium oxide nanoparticles is 2nm-10nm. Preferably, the exogenous mitochondria are derived from c57BL/6 mouse heart tissue. Preferably, the composite delivery system has a hydration kinetic diameter of 600nm to 900nm. The present invention next provides a method of preparing the composite delivery system, comprising the steps of: s1, preparing a thiolated modified cRGDyk targeting peptide (SH-cRGDyk); s2, preparing cerium oxide nano-particles (Mal-Ce NPs) with the surfaces modified by maleimide groups; s3, extracting mitochondria from heart tissues of the c57BL/6 mice, and carrying out sulfhydrylation modification on the mitochondria by using Traut' S reagent to obtain sulfhydrylation modified exogenous mitochondria (SH-Mito); s4, incubating the product obtained in the step S1 and the product obtained in the step S2 to obtain cRGDyk-cerium oxide nanoparticle compound (cRGDyk-Ce); and S5, incubating the compound obtained in the step S4 and the product obtained in the step S3 on ice to enable the residual maleimide groups on the surface of the compound to undergo a coupling reaction with sulfhydryl groups on the surface of mitochondria, and purifying to obtain the cRGDyk-mitochondria composite delivery system. The invention finally provides the use of the above-described composite system for the preparation of a medicament for the prevention and/or treatment of liver fibrosis. Preferably, the medicament is administered by intravenous injection. Compared with the prior art, the application has the followi