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Application of neuronal guidance factor Sema gene recombinant lentiviral vector in preparation of drug for treating osteoarthritis

A technology of lentiviral vectors and guiding factors, applied in the direction of viruses/bacteriophages, the use of vectors to introduce foreign genetic material, applications, etc., to achieve continuous and efficient effects, prevent and slow down the progress of osteoarthritis

Active Publication Date: 2020-01-14
WEST CHINA HOSPITAL SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] Although the prior art discloses the application of Sema3A for the treatment of bone-related diseases, it is only for the treatment of bone and does not involve cartilage. The present invention focuses on the treatment of articular cartilage

Method used

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  • Application of neuronal guidance factor Sema gene recombinant lentiviral vector in preparation of drug for treating osteoarthritis
  • Application of neuronal guidance factor Sema gene recombinant lentiviral vector in preparation of drug for treating osteoarthritis
  • Application of neuronal guidance factor Sema gene recombinant lentiviral vector in preparation of drug for treating osteoarthritis

Examples

Experimental program
Comparison scheme
Effect test

preparation example 1

[0032] Preparation example 1 Preparation of Sema 3a gene lentiviral recombinant vector

[0033] 1. Plasmid Linearization

[0034] Use Bbs1 to excise the shRNA fragment from the plasmid to obtain a linearized plasmid fragment.

[0035] 2. Integrate the Sema 3a gene into the plasmid

[0036] Mix the linearized plasmid fragment with the Sema 3a gene sequence of the target gene, and realize the connection of the plasmid and the target gene under the action of T4-DNA ligase.

[0037] 3. 293T cell culture

[0038] 293T cells are derived from the commonly used HEK293 cell line. HEK293 cells are a cell line obtained by transforming human embryonic kidney cells with sheared adenovirus type 5 DNA. On this basis, the cells transformed with the SV40T antigen are called 293T cells, which grow faster and have more vigorous metabolism.

[0039] 3.1 Recovery of 293T cells

[0040] 1) preheating culture medium;

[0041] 2) Take out the cell cryopreservation tube from the liquid nitrogen...

Embodiment 1

[0090] The preparation of the medicine of embodiment 1 Sema 3a gene lentiviral recombinant vector

[0091] A drug solution was prepared by dissolving 100 μL of the Sema 3a gene lentiviral recombinant vector solution prepared in Preparation Example 1 in 900 μL of PBS solution with pH=7.4.

Embodiment 2

[0092] The preparation of the medicine of embodiment 2 Sema 3a gene lentiviral recombinant vector

[0093] A drug solution was prepared by dissolving 100 μL of the Sema 3a gene lentiviral recombinant vector solution prepared in Preparation Example 1 in 900 μL of D-PBS solution with pH=7.4.

[0094] Effect example Therapeutic effect of Sema 3a gene lentiviral recombinant vector drug

[0095] 1. Purpose of the test: Taking C57 mice as a model, evaluate the therapeutic effect of the Sema 3a gene lentiviral recombinant vector drug prepared by the present invention on the mouse arthritis model.

[0096] 2. The test process is as follows:

[0097] ① Knee arthritis model establishment:

[0098] Animals: 30 8-week-old C57 mice were randomly divided into 3 groups, ACLT of the right leg: model of anterior cruciate ligament transection.

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Abstract

The invention discloses application and a preparation method of a neuronal guidance factor Sema gene recombinant lentiviral vector in preparation of a drug for treating osteoarthritis. A gene sequenceof neuronal guidance factor Sema is a complete sequence shown in SEQ ID NO:1, a sequence similar to a sequence shown in the SEQ ID NO:1 or a variant of the sequence shown in the SEQ ID NO:1. The drugprepared by the method can induce target tissue of organisms to highly express sema polypeptide with lentivirus as the vector after acting on the human body, can make a patient locally and highly express sema 3a protein for a long time, and has continuous and efficient functions.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to the application of a recombinant lentiviral vector of the nerve guiding factor Sema gene in the preparation of a medicine for treating osteoarthritis. Background technique [0002] Osteoarthritis (OA) refers to a synovial joint disease characterized by cartilage loss and periarticular bone reaction, also known as osteoarthritis, degenerative osteoarthritis, hypertrophic or hypertrophic arthritis, etc. Most common rheumatic disease worldwide. Osteoarthritis is the degenerative injury of articular cartilage, reactive hyperplasia of joint margins and subchondral bone caused by aging, obesity, strain, trauma, congenital abnormalities of joints, joint deformities and many other factors. Osteoarthritis is the most common joint disease among middle-aged and elderly people. According to survey research, the prevalence rate of people over 60 years old can reach 50%, and that of peo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/867C12N15/66A61K38/19A61P19/02A61P19/08
CPCC12N15/86A61K38/19A61K9/0014A61P19/02A61P19/08C12N2740/15043Y02A50/30
Inventor 黄石书宁宁陈佳丽龙丹吴迪伟尤炫合
Owner WEST CHINA HOSPITAL SICHUAN UNIV
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