CN-122005858-A - MKP7 targeted bionic nanoparticle preparation and preparation method and application thereof

Abstract

The invention discloses a MKP7 targeted bionic nanoparticle preparation and a preparation method and application thereof. The MKP7 targeted bionic nanoparticle preparation is pHLIP modified macrophage membrane coated bionic nanoparticle siMKP7@NP-M-pHLIP loaded with siMKP. The preparation takes PLGA-PEI copolymer as a carrier core to wrap siMKP, coats a macrophage membrane on the outer layer, and modifies pHLIP targeting peptide on the surface of the membrane to construct spherical nanoparticles with bionic camouflage and pH sensitive targeting delivery functions. In-vitro experiments show that the nanoparticle can be effectively ingested by macrophages and can obviously inhibit ox-LDL induced macrophage lipid deposition, in-vivo experiments prove that the preparation can be targeted and delivered to an atherosclerosis plaque part, and the preparation can effectively reduce carotid plaque area, reduce serum inflammatory factor level and regulate blood lipid metabolism by inhibiting MKP7 expression in macrophages, thereby inhibiting atherosclerosis development and providing a high-efficiency and low-toxicity nucleic acid drug delivery system for atherosclerosis treatment.

Inventors

• MA JIE
• JIAO PING
• WANG WEI
• LI MINGMING
• ZHAO JIANAN
• HOU KEYI

Assignees

• 吉林大学

Dates

Publication Date

20260512

Application Date

20260416

Claims (9)

1. 1. An MKP7 targeted bionic nanoparticle preparation for treating atherosclerosis, which is characterized in that the MKP7 targeted bionic nanoparticle preparation is pHLIP modified macrophage membrane coated siMKP-NP nanoparticle siMKP7@NP-M-pHLIP, and siMKP comprises a nucleotide sequence shown as SEQ ID NO: 1.
2. 2. The MKP7 targeted biomimetic nanoparticle formulation of claim 1, wherein pHLIP comprises the amino acid sequence shown in SEQ ID No. 2.
3. 3. The MKP7 targeted biomimetic nanoparticle formulation of claim 1, wherein the macrophage membrane is a cell membrane isolated from primary macrophages and macrophages of cell line origin.
4. 4. The MKP7 targeted biomimetic nanoparticle formulation of claim 1, wherein the primary macrophage membrane comprises a bone marrow-derived macrophage membrane, an abdominal cavity macrophage membrane, and a human peripheral blood mononuclear macrophage membrane and an embryo-derived tissue resident macrophage membrane, the cell line-derived macrophage membrane comprises a human macrophage line THP-1 cell membrane, a mouse macrophage line RAW264.7 cell membrane, and a mouse mononuclear macrophage J774 cell membrane.
5. 5. The MKP7 targeted biomimetic nanoparticle formulation of claim 1, wherein the MKP7 targeted biomimetic nanoparticle formulation is of spherical structure.
6. 6. The MKP7 targeted biomimetic nanoparticle formulation of claim 1, wherein the MKP7 targeted biomimetic nanoparticle formulation is an injection.
7. 7. A method for preparing a MKP7 targeted biomimetic nanoparticle formulation, for use in preparing the MKP7 targeted biomimetic nanoparticle formulation according to any one of claims 1 to 6, comprising the steps of: Preparing siMKP7@NP nanoparticles, namely taking a synthetic PLGA-PEI copolymer as a raw material of a nano carrier, and wrapping siMKP < 7 > into the nanoparticles to prepare the siMKP7@NP; Preparing siMKP7@NP-M nanoparticles, namely separating to obtain cell membranes of macrophages after obtaining the macrophages, further preparing to obtain cell membrane vesicles, and wrapping the cell membrane vesicles on the surfaces of the siMKP7@NP nanoparticles to obtain the siMKP7@NP-M nanoparticles; Preparation of siMKP7@NP-M-pHLIP nanoparticles SiMKP7@NP-M nanoparticles were mixed with pHLIP to prepare siMKP7@NP-M-pHLIP nanoparticles.
8. Use of mkp7 as a drug target in the preparation of a formulation for the treatment of atherosclerosis.
9. 9. The use according to claim 8, wherein the formulation treats atherosclerosis by inhibiting the expression or activity of MKP7 in macrophages, the formulation comprising siMKP7, the siMKP comprising the nucleotide sequence set forth in SEQ ID NO: 1.

Description

MKP7 targeted bionic nanoparticle preparation and preparation method and application thereof Technical Field The invention relates to the technical field of biological medicines, in particular to a MKP7 targeted bionic nanoparticle preparation and a preparation method and application thereof. Background Atherosclerosis (AS) is a major pathological basis of cardiovascular diseases (such AS coronary heart disease, myocardial infarction and cerebral apoplexy) and is caused by plaque formation caused by deposition of substances such AS lipid, cholesterol and the like on the inner wall of an arterial blood vessel, thickening and hardening of the blood vessel wall and stenosis of a lumen, and finally, blood flow supply is influenced, so that immune chronic progressive diseases are caused. The main causes of AS include vascular endothelial injury caused by factors such AS hypertension, high cholesterol, smoking, diabetes, etc., lipid deposition caused by lipid accumulation such AS low density lipoprotein cholesterol (LDL-C), etc., inflammatory reaction caused by immune cell accumulation, etc., and in addition, aging, unhealthy diet, smoking, lack of exercise, obesity, diabetes, etc. are also risk factors for causing AS lesions. Current therapeutic strategies for AS mainly include lifestyle interventions, drug therapies, and interventional and surgical therapies. Among the drugs used in AS treatment are mainly statin drugs for reducing LDL-C content, stabilizing plaque, antiplatelet drugs for preventing thrombosis, and antihypertensive/hypoglycemic drugs for controlling blood pressure and blood sugar. However, the existing therapeutic strategies have a certain limitation, which fails to consider the biological complexity of AS, and conventional lipid-lowering statin therapy and the like cannot be directed to various mechanisms driving plaque formation, and thus, there is an urgent need to develop a new formulation for AS treatment. Mitogen-activated protein kinases (MAPKs) are a class of serine/threonine protein kinases that are involved in regulating growth, differentiation and proliferation of cells, and in regulating cell cycle and apoptosis. MAPK phosphatases (Mitogen kinase phosphatases, MKPs), also known as bispecific phosphatases (Dual specificity phosphatase, DUSP), can down-regulate MAPK by dephosphorylating tyrosine/threonine residues. MKP7 is an important bispecific phosphatase, one of the members of the MKPs family, playing a key role in cell signaling, inflammatory regulation and tumorigenesis. MKP7 plays an important role in metabolism and diabetes, such as enhancing MKP7 expression can inhibit ROS or JNK activation induced by endoplasmic reticulum stress, protecting islet beta cell survival, and MKP7 relieves liver steatosis induced by metabolic stress by negatively regulating JNK signals, and MKP7 regulates congenital and adaptive immune responses by inhibiting JNK/p38 pathways in the immune system, and MKP7 deficiency leads to overactivation of CD4+ T cells, promoting Th17 cell differentiation. At present, the mechanism of action of MKP7 in AS is not yet clear. The invention reveals the key role of MKP7 in AS development for the first time, and intervenes by taking MKP7 AS a target point. Nanoparticles (NPs) are formulations composed of a wide variety of molecular compounds, which are generally structurally complex, with a complex array of functions that can facilitate diagnosis and treatment of AS. Unlike traditional drug therapies that have only a single specific therapeutic function, engineered multifunctional NPs can be designed with the ability to sense and deliver a pharmacological activator, inhibitor, or genetic material load. NPs were originally designed for the treatment of cancer, and also have a high AS potential due to the high overlap of tumors with the biological mechanisms of AS plaque formation, such AS risk factors, inflammation, and the action of macrophages. The nanocarriers for delivering nucleic acids mainly include liposomes, viruses, and biomimetic nanoparticles. The virus vector has higher transfection efficiency and is widely used for delivering miRNA, however, the virus vector has great hidden danger in the aspect of safety, further clinical application is limited, the problems of lower targeting property of the lipid vector, certain immunogenicity and the like are not solved at present, and the bionic nanoparticle is a recently developed nano vector with lower immunogenicity and higher biocompatibility, so that the MKP 7-loaded targeted bionic nanoparticle is to be developed for AS treatment. As foreign matter, the nanoparticles are easily recognized and removed by the immune system, and it is difficult to reach the lesion. In recent years, bionic nanoparticles are becoming research hotspots in the tumor-related field with unique advantages, and the bionic nanoparticles mainly construct a drug delivery system by simulating or directly utilizing natural cell components (s