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External compound formula for comprehensively preventing and treating epidermal stem cell reactive oxygen species damage based on apocynin

A technology of epidermal stem cells and reactive oxygen species, applied in the field of external compound formulations, can solve the problems of affecting the natural repair ability of the skin, not fully considering ROS, interfering with the normal function of cells, etc., to alleviate ROS damage, strong innovation, and reduce NADPH Effect of oxidase activity

Inactive Publication Date: 2020-07-31
陕西哲联生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The generation and existence of ROS has an important difference between the cell region and the polar and non-polar phase. The elimination of ROS also has two modes: metabolic free energy drive and reaction itself free energy drive. Covering the main aspects of ROS damage, for example, the application numbers 201710793731.X, 201610719836.6 and 201680013312.3 did not fully consider the relationship between the three main pathways of ROS metabolism, which led to the simple elimination of ROS to interfere with the normal function of cells, but instead affected the natural repair ability of the skin; another application No. 201710733106.6, 201410804802.8 and 201710126577.0 did not target the key mechanism of ROS free radical-induced lipid peroxidation damage to the integrity of cell membrane structure, resulting in ineffective prevention and treatment of ROS-induced lipid peroxidation, a key aging mechanism

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1 Evaluation of formulation effectiveness and safety based on high-throughput standardized cell experiments.

[0028] On the basis of previous work, using high-throughput standard cell experiments, the optimal dosage of each component of the formula was screened out to form formula A: Apocynum apocynum (0.1%), magnesium ascorbyl phosphate (0.8%), tert-butyl Hydroquinone (0.2%) and Dihydrolipoic Acid (0.4%).

[0029] Based on the previous work of the present invention, select the corresponding ratio concentration of each component in formula A respectively (the mass concentration ratio is according to the above-mentioned order: 1:8:2:4, with Apocynum apocynum in culture medium final concentration 10 μ mol / The formula A configured according to the above-mentioned ratio is 1 U) at the time of L), and the cell viability was measured by the CCK8 method, and the T test analysis was carried out on the basis of the data of 5 independent experiments; with P0.01 is used ...

Embodiment 2

[0030] Example 2 ROS free radical scavenging effect experiment of epidermal stem cells.

[0031] Based on the ratio and final concentration units of formula A in Example 1, the intracellular ROS elimination and damage effects were determined at an optimal concentration of 20 U. First, DCF probe was used to measure the relative concentration of intracellular ROS, and it was found that the A pretreatment group was 51.7% lower than the control group; secondly, the cell death rate was measured by propidium iodide, and it was found that the formula A pretreatment group had a mortality rate of 0.7%, and the negative control group was 3.6%. %, after 5 independent experiments, the T test found that the difference was significant. The above data shows that this formula A can reduce the level of intracellular ROS and reduce cell death, so it can effectively protect the cell damage caused by ROS.

Embodiment 3

[0032] Example 3 A randomized double-blind controlled multi-center volunteer experiment of this formula at an optimal concentration.

[0033] From January 2018 to April 2019, in Beijing, Guangzhou and Xianyang, a total of 150 volunteers were selected from each city, with an average age of 36.1 years, a male to female ratio of 0.32, and a body mass index of 20.81. Volunteers used formula A, marketed product B and marketed product C prepared in Example 1, which were indistinguishable in appearance, for a total of 120 days each day (formula A was dosed according to Example 1, and products B and C were taken according to the instructions), Volunteer experiments were conducted using the principle of randomized double-blind control.

[0034] The skin aging indicators of the volunteers: skin thickness, skin mechanical change (Ballistometry method), paper pattern change, wrinkle change and uneven pigmentation (pigmentation index and erythema index) were comprehensively scored. The res...

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Abstract

The invention discloses an external compound formula for comprehensively preventing and treating epidermal stem cell reactive oxygen species damage based on apocynin. The external compound formula comprises the following components of (0.002-0.4%) of apocynin, (0.2-5%) of magnesium ascorbyl phosphate, (0.05-0.8%) of tert-butylhydroquinone and (0.1-3%) of dihydrolipoic acid. The inventor establishes a theoretical framework of improving the ROS proliferation promoting second messenger sensitivity of cells, reducing the Nox enzyme activity and relieving the ROS damage of epidermal stem cells on the basis of regulating mitochondrial respiratory chain and lysosomal ROS production in long-term research; on the basis, the technical scheme of low-activity Nox enzyme with apocynin as the main component is screened in a targeted manner, so that ROS senescence injury is comprehensively relieved. In addition, in formula development, a high-throughput standardized cell experiment and a random double-blind control multi-center volunteer experiment are utilized to accurately detect the action range and toxicity threshold of each component, so that the dosage has a certain tolerance level and extremely high safety, and the formula is suitable for being used as a core formula for product development.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to an external compound formula for comprehensively preventing and treating the damage of active oxygen clusters of epidermal stem cells based on apocynum. Background technique [0002] The skin is the largest organ in the human body, accounting for about 14% of the body weight. It has important functions such as protection, excretion, temperature regulation, and sensing external stimuli. It can be divided into two layers: the dermis and the epidermis. The epidermis is located in the outermost layer and is mainly composed of basal layer, granular layer, transparent layer and stratum corneum, among which some cells in the basal layer have high differentiation potential, and have slow periodicity, adhesion and strong self-renewal and differentiation capabilities, etc. Typical characteristics of stem cells, so they are called epidermal stem cells (ESCs). [0003] Epidermal stem cells prolif...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/12A61K31/661A61K31/385A61K31/05A61P17/18A61P17/00
CPCA61K31/05A61K31/12A61K31/385A61K31/661A61P17/00A61P17/18A61K2300/00
Inventor 王文娟贺小星
Owner 陕西哲联生物科技有限公司
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