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Application of alpha-ketoglutaric acid in preparation of medicine

A kind of ketoglutaric acid, a technology for preparing medicines, applied in the biological field, to achieve the effect of promoting the regeneration of skin hair follicles, promoting a significant increase, and preventing the progression of osteoarthritis

Pending Publication Date: 2021-06-25
TONGJI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, there is no report of α-KG in the treatment of arthritis. Therefore, providing a drug containing α-KG has great potential application value for the clinical treatment of osteoarthritis

Method used

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  • Application of alpha-ketoglutaric acid in preparation of medicine
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  • Application of alpha-ketoglutaric acid in preparation of medicine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] Embodiment 1 The cultivation of mouse chondrocytes

[0067] Mouse chondrocytes were obtained from 7d-old C57BL / 6 mice (Shanghai Slack Animal Co., Ltd.). Neutral collagenase (Serva, Germany) was digested for 8 h, and the cell pellet was obtained by centrifugation. After that, the cells were resuspended with 10% FBS-containing modified Eagle medium (DMEM) (Gibco, USA) culture medium and inoculated in a culture dish at 37°C. , 5% CO2 constant temperature incubator, cultured with high-sugar DMEM medium (containing 10% FBS, 100U / mL penicillin and 100mg / L streptomycin). The medium was changed for the first time at 48 hours. When the cell fusion rate reached 80%-90%, it was digested and passaged with 0.25% trypsin (Gibco, USA). According to this method, the medium was changed and subcultured, and the obtained cells were respectively recorded as P1, P2, P3 generation and so on.

Embodiment 2I

[0068] Example 2 Effect of IL-1β on glutamine content levels in chondrocytes cultured in glutamine-containing medium

[0069] 1 Experimental method

[0070] (1) Preparation of standard solution

[0071] Accurately weigh nineteen kinds of amino acid standards, and prepare a mixed solution with water to make the final concentration 0.05μg / mL, 0.1μg / mL, 0.2μg / mL, 0.5μg / mL, 1μg / mL, 2μg / mL, 5μg / mL, 10μg / mL, 20μg / mL, 50μg / mL, 100μg / mL eleven standard concentration gradients.

[0072] (2) Sample pretreatment

[0073] Take the sample and vortex for 30s, freeze it quickly in liquid nitrogen, dissolve the quenched cell solution at room temperature, vortex for 30s, centrifuge at 800g for 1min, take the supernatant into a centrifuge tube, and place it on dry ice; use 500μL methanol (-80℃) Reset the cells, place them in liquid nitrogen for rapid freezing, repeat the vortex and centrifugation steps, combine the supernatants, dry them with liquid nitrogen, add 1000 μL of alanine-d4 isoto...

Embodiment 3I

[0086] Example 3 Effects of IL-1β on the decomposition phenotype and synthesis phenotype of chondrocytes cultured without glutamine

[0087] 1 Experimental method

[0088] (1) Treatment of mouse chondrocytes

[0089] The P1 generation mouse chondrocytes cultured in Example 1 were used, and after the P1 generation chondrocytes grew adherently, they were treated with serum-free DMEM culture medium with 10 ng / ml IL-1β. For chondrocytes treated without glutamine, the culture medium without glutamine was changed 12 hours in advance, and then chondrocytes were treated with IL-1β.

[0090] (2) Real-time fluorescent quantitative PCR (Real Time q-PCR) analysis

[0091] 1) Using the P1 generation mouse chondrocytes cultured in Example 1, suck out the culture medium in the culture dish and wash it once with PBS.

[0092] 2) Extraction of Total RNA

[0093] ① Add chloroform (1 / 5 volume of RNAiso Plus) to the homogenate lysate, close the cap of the centrifuge tube tightly, shake vigoro...

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Abstract

The invention relates to the technical field of biology, in particular to an application of alpha-ketoglutaric acid in preparation of drugs, and the drugs are used for 1) treating osteoarthritis and related diseases; and / or, 2) inhibiting the decomposition phenotype of cartilage cells; and / or 3) promoting synthetic phenotypes of cartilage cells; and / or 4) promoting skin hair follicle regeneration. The pharmaceutical composition provided by the invention can protect cartilage tissues and prevent the progress of osteoarthritis, and has great potential application value for clinical treatment of osteoarthritis. In addition, the medicine provided by the invention can promote obvious increase of skin hair follicles, and also has great potential application value in treatment and improvement of alopecia.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the use of alpha-ketoglutarate in the preparation of medicines. Background technique [0002] Osteoarthritis (osteoarthritis osteoarthritis, OA) is a common chronic joint disease. Osteoarthritis has a high prevalence rate and ranks sixth in the world for disabling diseases. It is expected that by 2020, it will rise to the fourth place among disabling diseases. The onset is more common in middle-aged and elderly people, and women are more common than men. It mainly occurs in the heavy-bearing knee joints, ankle joints, spine and frequently moving finger joints. The main pathological change of osteoarthritis is the degeneration of articular cartilage, and the clinical symptoms usually include joint pain, tenderness, stiffness, and functional limitation of osteoarthritis, accompanied by synovitis and secondary osteophyte formation. Osteoarthritis causes destruction of articular cart...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/194A61P29/00A61P19/02A61P19/04A61P19/08A61P17/14
CPCA61K31/194A61K9/0019A61P29/00A61P19/02A61P19/04A61P19/08A61P17/14
Inventor 崔磊李帅军
Owner TONGJI UNIV
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