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Compound formula for relieving testis senescence injury by comprehensively correcting cell reactive oxygen species metabolic disorders

A technology of metabolic disorder and cell activity, which is applied in the direction of pharmaceutical formulations, organic active ingredients, medical preparations containing active ingredients, etc., can solve the problems of not considering the role of second messengers, spermatogenesis inhibition, and large side effects, etc., to benefit the product Develop and market, improve the level of expression, increase the effect of the effect

Inactive Publication Date: 2020-09-08
NORTHWEST A & F UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] At present, the prevention and health care programs for relieving ROS aging damage of testicular cells generally have problems such as large side effects or poor effects. Oxidative, promotes the elimination of ROS, but the above-mentioned patent does not consider the second messenger function of ROS itself, and simply eliminates intracellular ROS, which may seriously interfere with the effect of ROS as a second messenger to promote the proliferation of spermatogonial stem cells in the testis, and there is a risk of spermatogenesis inhibition Patents such as application number 201610218973.1 only propose the target of drug action, and there is no mature and effective drug molecule that can target this target; patents such as application number 201911011122.X use a matrine derivative and find that the compound has anti- The apoptosis function of radiation cells and the effect of inhibiting the production of ROS, nitric oxide (NO) and inflammatory factors in radiation-damaged cells, but it can only protect radiation damage has the problem that the scope of action is too limited. Generally speaking, the testis is not easy to receive sunlight ultraviolet rays Radiation damage of the testis is almost only seen in people who receive radiotherapy and special occupations, so the scope of application of this patent is limited and cannot be used as a widely used technical solution for pathological aging of the testis
In conclusion, there is currently no safe and effective mature solution for the prevention and treatment of testicular pathological aging by targeting ROS damage in testicular cells

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1 Evaluation of formulation effectiveness and safety based on high-throughput standardized cell experiments.

[0034] 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: melatonin (2mg / d), lipoic acid (200mg / d), nicotinamide riboside (300mg / d).

[0035] Based on the previous work of the present invention, the corresponding ratio concentrations of each component in the formula A were respectively selected (the mass concentration ratio is according to the above order: 1:100:150, when the final concentration of melatonin in the culture solution is 0.1 μmol / L The formula A configured according to the above ratio is 1 U), and the viability and ATP production of vascular endothelial cells were measured by the absorbance method, and the T test analysis was carried out on the basis of the data of 5 independent experiments; with P0.01 is used as the toxic d...

Embodiment 2

[0036] Example 2 Experiment on the elimination effect of ROS free radicals in testicular cells.

[0037] 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 50 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 negative control group; secondly, the cell death rate was measured by propidium iodide staining flow cytometry, and it was found that the formula A pretreatment group had a mortality rate of 0.8 %, the negative control group was 7.9%, and the T test found that the difference was significant after 5 independent experiments. 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

[0038] Example 3 A randomized double-blind controlled multi-center volunteer experiment of this formula at an optimal concentration.

[0039] From April 2018 to April 2019, in Beijing, Xi'an and Xianyang, a total of 150 male volunteers were selected from each city, with an average age of 41.2 years and a body mass index of 22.3. Volunteers used formula A prepared in Example 1 whose appearance could not be distinguished, and products B and C already on the market for a total of 120 days (formula A was dosed according to Example 1, and products B and C were taken according to the instructions). Volunteer experiments were conducted on the principle of blind control.

[0040] The sperm count, motility, acrosome integrity and sperm ROS concentration of the volunteers were measured respectively, and the above results were normalized to generate a sperm pathological aging damage score. The results show that formula A can finally reduce the pathological aging damage of volunteers' sp...

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Abstract

The invention discloses a compound formula for relieving testis senescence injury by comprehensively correcting cell reactive oxygen species (ROS) metabolic disorders. The compound formula comprises the following main components: melatonin (0.5-3mg / d), lipoic acid (50-500mg / d) and nicotinamide ribose (50-700mg / d). The components in the formula have clever metabolic synergy: the nicotinamide ribosepromotes generation of a main carrier of reduction potential and free energy, so that the lipoic acid is used as a redox switch to fully eliminate ROS damage of testicular cells; NADPH oxidase generated by nicotinamide ribose metabolism is catalyzed to generate second messenger ROS for maintaining self-renewal of testicular spermatogonial stem cells, and side effects can be ingeniously avoided when the lipoic acid and the melatonin eliminate ROS and interfere spermatogenesis. According to the formula, a high-flux standard cell experiment and a random double-blind control multi-center volunteer experiment are utilized to accurately ascertain the effects and toxicity threshold values of the components of the formula, so that the use dosages of the components have extremely high tolerance and safety, and the formula is suitable for being used as a core formula for development of related products.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a composite formula for alleviating testicular aging damage by comprehensively correcting the metabolic disorder of active oxygen species in cells. Background technique [0002] With the increasing aging of our country's population, aging and geriatric diseases have become a huge challenge. In addition to causing hypofertility, the aging of the gonads, accompanied by endocrine disorders such as low androgen levels, will cause the decline of multiple systems and organ functions throughout the body. The testis has two functions of producing sperm and secreting androgen, so the aging of the testis will cause the decline of spermatogenesis and fertility, but more importantly, the lack of androgen caused by the aging of the testis can cause sexual dysfunction, osteoporosis, Systemic symptoms such as obesity and metabolic syndrome may also lead to neurological and mental symptoms such as de...

Claims

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Application Information

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IPC IPC(8): A61K31/4045A61K31/385A61K31/706A61P15/08A61P39/06
CPCA61K31/4045A61K31/385A61K31/706A61P15/08A61P39/06A61K2300/00
Inventor 贺鑫
Owner NORTHWEST A & F UNIV
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