Application of resorcylic acid lactone 5Z-7 in preparation of medicament for treating osteoarthritis

Abstract

The invention discloses application of resorcylic acid lactone 5Z-7 in preparation of a medicament for treating osteoarthritis and belongs to the field of biological medicines. The resorcylic acid lactone 5Z-7 is a small-molecule inhibitor for TAK1, and activation of immuno-associated pathway, such as NF-kB, p38 and JNK mediated by inflammatory cytolines and a matrix degradation product in synovium and cartilage tissue can be inhibited through targeted inhibition of the enzymatic activity of the TAK1 in an articular cavity, thereby achieving the targets of reducing synovial inflammatory response, inhibiting cartilage degradation and improving the microenvironment of the articular cavity through controlling a series of gene expression related to the osteoarthritis on the downstream of the pathways; the resorcylic acid lactone 5Z-7 is of great significance in delaying the osteoarthritis process and effectively treating the osteoarthritis. In addition, the resorcylic acid lactone 5Z-7 is taken as a small molecule compound, the action target is single and high in specificity, and the resorcylic acid lactone 5Z-7 is free of a toxic or side effect on cartilage tissue through detection, and is safer and more reliable.

Description

technical field

[0001] The invention relates to the field of biomedicine, in particular to the application of a dihydroxybenzoic acid lactone 5Z-7 in the preparation of medicines for treating osteoarthritis. Background technique

[0002] Osteoarthritis (OA) is a common chronic and degenerative joint disease. Its clinical manifestations are slowly developing joint pain, tenderness, stiffness, joint swelling, limited mobility and joint deformity, which seriously interfere with the daily life of patients. Life. Therefore, timely and effective treatment of osteoarthritis is necessary.

[0003] The existing technologies for the treatment of osteoarthritis mainly include non-drug treatment and drug treatment. Generally speaking, non-drug treatment includes weight reduction, moderate exercise, physical therapy and surgical treatment. The physical therapy cycle is long and the effect is slow, and surgical treatment can Choose arthroscopic joint debridement, osteotomy and artificia...

Examples

Embodiment 1

[0064] This example clarifies that the overexpression of TAK1 in the rat joint cavity can cause pathological changes related to osteoarthritis, and the specific operation steps are as follows:

[0065] Fifteen male SD rats with a body weight of about 100 g were randomly divided into three groups, 5 rats in each group, and the rats in each group were injected with control empty adenovirus, 1×10 7 pfu dose of TAK1 adenovirus and 1×10 8 The pfu dose of TAK1 adenovirus corresponds to the empty adenovirus control group, the low-dose TAK1 adenovirus group and the high-dose TAK1 adenovirus group (wherein, pfu represents the quantitative unit of adenovirus). The rats were sacrificed after the injection was continued twice a week for one month. The articular cartilage of the rats in each group was taken out, and the general photos were taken, and then they were fixed in paraformaldehyde with a mass concentration of 4% for 48 hours, and then placed in decalcification solution for 48 ho...

Embodiment 2

[0072] This example clarifies the effect of the TAK1 inhibitor dihydroxybenzoic acid lactone 5Z-7 (hereinafter referred to as 5Z-7) on the proliferation of chondrocytes and synoviocytes, as well as its effect on the activity of NF-κB and MAPK pathways. The specific operation The steps are as follows:

[0073] 1. CCK-8 method (Cell Counting Kit-8) to detect the effect of 5Z-7 on the activity of chondrocytes and synoviocytes

[0074] Chondrocytes and synovial cells from patients with osteoarthritis were isolated and cultured, as well as primary chondrocytes from normal rats. 4 Inoculated into each well of a 96-well plate at a density of 200 μL, placed in a cell culture incubator for pre-culture for 24 hours (at a temperature of 37°C, with a volume fraction of 5% CO 2 ), then sequentially add 0, 0.1, 0.2, 0.3, 0.5, 1, 2, 3 μmol / L of 5Z-7 (6 replicate wells for each concentration), and add 10 μL of CCK-8 solution to each well after culturing for 24 hours , place the culture plat...

Embodiment 3

[0093] This example clarifies the regulatory effect of the TAK1 inhibitor 5Z-7 on the expression and regulation of osteoarthritis-related genes in chondrocytes and synoviocytes. The specific operation process is as follows:

[0094] 1) To clarify the regulatory effect of 5Z-7 on the expression of osteoarthritis-related genes in chondrocytes

[0095] In this example, experiments were carried out on the effect of 5Z-7 on the expression of matrix degrading enzymes, inflammatory factors and extracellular matrix components in chondrocytes. Specifically, IL-1β was added to normal rats alone or together with 5Z-7 In the culture supernatant of primary mouse chondrocytes, after 2 hours and 4 hours of treatment, the cell pellet was collected with Trizol, and the expression of osteoarthritis-related genes in the cells of each group was detected by real-time fluorescent quantitative PCR instrument. Among them, matrix degrading enzymes include: MMP1, MMP13, ADAMTS1, ADAMTS5, inflammatory f...

technical field

[0001] The invention relates to the field of biomedicine, in particular to the application of a dihydroxybenzoic acid lactone 5Z-7 in the preparation of medicines for treating osteoarthritis. Background technique

[0002] Osteoarthritis (OA) is a common chronic and degenerative joint disease. Its clinical manifestations are slowly developing joint pain, tenderness, stiffness, joint swelling, limited mobility and joint deformity, which seriously interfere with the daily life of patients. Life. Therefore, timely and effective treatment of osteoarthritis is necessary.

[0003] The existing technologies for the treatment of osteoarthritis mainly include non-drug treatment and drug treatment. Generally speaking, non-drug treatment includes weight reduction, moderate exercise, physical therapy and surgical treatment. The physical therapy cycle is long and the effect is slow, and surgical treatment can Choose arthroscopic joint debridement, osteotomy and artificia...

Examples

Embodiment 1

[0064] This example clarifies that the overexpression of TAK1 in the rat joint cavity can cause pathological changes related to osteoarthritis, and the specific operation steps are as follows:

[0065] Fifteen male SD rats with a body weight of about 100 g were randomly divided into three groups, 5 rats in each group, and the rats in each group were injected with control empty adenovirus, 1×10 7 pfu dose of TAK1 adenovirus and 1×10 8 The pfu dose of TAK1 adenovirus corresponds to the empty adenovirus control group, the low-dose TAK1 adenovirus group and the high-dose TAK1 adenovirus group (wherein, pfu represents the quantitative unit of adenovirus). The rats were sacrificed after the injection was continued twice a week for one month. The articular cartilage of the rats in each group was taken out, and the general photos were taken, and then they were fixed in paraformaldehyde with a mass concentration of 4% for 48 hours, and then placed in decalcification solution for 48 ho...

Embodiment 2

[0072] This example clarifies the effect of the TAK1 inhibitor dihydroxybenzoic acid lactone 5Z-7 (hereinafter referred to as 5Z-7) on the proliferation of chondrocytes and synoviocytes, as well as its effect on the activity of NF-κB and MAPK pathways. The specific operation The steps are as follows:

[0073] 1. CCK-8 method (Cell Counting Kit-8) to detect the effect of 5Z-7 on the activity of chondrocytes and synoviocytes

[0074] Chondrocytes and synovial cells from patients with osteoarthritis were isolated and cultured, as well as primary chondrocytes from normal rats. 4 Inoculated into each well of a 96-well plate at a density of 200 μL, placed in a cell culture incubator for pre-culture for 24 hours (at a temperature of 37°C, with a volume fraction of 5% CO 2 ), then sequentially add 0, 0.1, 0.2, 0.3, 0.5, 1, 2, 3 μmol / L of 5Z-7 (6 replicate wells for each concentration), and add 10 μL of CCK-8 solution to each well after culturing for 24 hours , place the culture plat...

Embodiment 3

[0093] This example clarifies the regulatory effect of the TAK1 inhibitor 5Z-7 on the expression and regulation of osteoarthritis-related genes in chondrocytes and synoviocytes. The specific operation process is as follows:

[0094] 1) To clarify the regulatory effect of 5Z-7 on the expression of osteoarthritis-related genes in chondrocytes

[0095] In this example, experiments were carried out on the effect of 5Z-7 on the expression of matrix degrading enzymes, inflammatory factors and extracellular matrix components in chondrocytes. Specifically, IL-1β was added to normal rats alone or together with 5Z-7 In the culture supernatant of primary mouse chondrocytes, after 2 hours and 4 hours of treatment, the cell pellet was collected with Trizol, and the expression of osteoarthritis-related genes in the cells of each group was detected by real-time fluorescent quantitative PCR instrument. Among them, matrix degrading enzymes include: MMP1, MMP13, ADAMTS1, ADAMTS5, inflammatory f...

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