阅读说明：本技术 烟碱酸衍生物在缓解女性生殖系统衰老损伤中的应用 (Application of nicotinic acid derivative in relieving female reproductive system aging injury ) 是由 贺鑫 于 2020-08-27 设计创作，主要内容包括：女性生殖系统衰老作为女性衰老的关键问题,其核心是细胞衰老,活性氧簇代谢紊乱广泛损伤生物大分子等,是细胞病理性衰老损伤主要分子机制。发明人经过长期实验筛选验证,发现烟酰胺、烟酸与烟酰胺单核苷酸等多种烟碱酸基衍生物均可提高胞内NADH和DNADP水平,矫正活性氧簇代谢紊乱,进而提高女性生殖系统细胞抗病理性氧化损伤的能力。还发现多种烟碱酸基衍生物仅存在药代动力学等方面的不同,一定程度上可以互相替代。在此基础上本发明公开了一种通过烟碱酸基衍生物通过综合矫正活性氧簇代谢紊乱缓解女性生殖系统衰老损伤在药物、保健品或功能食品方面的应用。(The female reproductive system aging is a key problem of female aging, and the key points of the female reproductive system aging are cell aging, active oxygen cluster metabolic disturbance widely damages biomacromolecules and the like, and the main molecular mechanism of cell pathological aging damage. The inventor finds that various nicotinic acid-based derivatives such as nicotinamide, nicotinic acid and nicotinamide mononucleotide can improve intracellular NADH and DNADP levels and correct metabolic disorder of active oxygen clusters through long-term experimental screening verification, and further improves the capacity of female reproductive system cells to resist pathological oxidative damage. It has also been found that various nicotinic acid-based derivatives differ only in terms of pharmacokinetics and the like and can be substituted for each other to some extent. On the basis, the invention discloses application of nicotinic acid-based derivatives in the aspects of medicines, health-care products or functional foods for relieving female reproductive system aging injury by comprehensively correcting active oxygen cluster metabolic disturbance.)

1. The invention provides a biotechnological scheme for comprehensively correcting metabolic disorders through nicotinic acid derivatives to relieve female reproductive system aging injury.

2. The nicotinic acid derivative as claimed in claim 1 refers to compounds containing nicotinoyl-like skeleton including but not limited to nicotinamide, nicotinic acid, nicotinamide riboside, nicotinic acid riboside and nicotinamide mononucleotide.

3. The compound of claim 1, which is administered primarily through the digestive tract, including but not limited to pharmaceuticals, nutraceuticals, and functional foods, but not including intravenous, intramuscular, topical, and aerosol inhalation.

Technical Field

The invention belongs to the technical field of biology, and relates to an application of a medicine, a health-care product or a functional food for comprehensively correcting metabolic disorder and relieving female reproductive system aging injury through a nicotinic acid derivative.

Background

With the increasing aging of the population in China, the health problem caused by aging becomes a great challenge at present. In addition to the decline of fertility, the aging of female reproductive system causes endocrine disorders such as low estrogen and progestogen levels, which causes the decline of functions of various systems and organs of the whole body, and may also cause nervous system and psychopsychological symptoms such as energy decline, memory decline, sleep disorder, depression, and the like, and even is closely related to the occurrence and development of serious chronic diseases such as diabetes, cardiovascular and cerebrovascular diseases, and the like, so the aging of female reproductive system is one of the core problems of female aging prevention and treatment, and needs to be paid high attention.

The heart of organ senescence is cell senescence, and telomere shortening, DNA damage, oxidative stress, and mitochondrial dysfunction are major causes of cell senescence. Cellular senescence is manifested by proliferation arrest and physiological function decline, and also secretes a series of cytokines, including proinflammatory factors, growth factors, chemokines, matrix remodeling enzymes and the like, which are called senescence-associated secretory phenotypes, thereby worsening local microenvironment, causing vicious circle and severely accelerating senescence process.

The progressive decline in the structure and function of the female reproductive system with age is a normal physiological process, but aging involves not only age-related physiological aging, but also premature aging and pathological aging due to various pathogenic factors, the latter being critical in prevention and treatment. Wherein telomeres are the aging clock built in the cell, so that replicative telomere shortening can be used as an indicator of physiological aging; however, activation of telomerase may lead to serious risks such as canceration of cells, and thus is not a good therapeutic target. DNA damage, oxidative stress and mitochondrial dysfunction are reasons causing pathological aging, wherein abnormal generation of Reactive Oxygen Species (ROS) metabolism is the root cause of the pathological aging, and ROS is the root cause of the oxidative stress, the DNA damage and the mitochondrial dysfunction, so that the prevention and treatment of the pathological aging are started from regulating and optimizing intracellular ROS level, and the problem is fundamentally solved.

Superoxide anions leaked by a mitochondrial respiratory chain are a main source of ROS (reactive oxygen species) of cells, during the aging process of a female reproductive system, the increased ROS leakage of the respiratory chain can damage the mitochondria in turn and aggravate the ROS leakage, and the positive feedback process is an important mechanism of pathological aging damage; because superoxide anion can be catalyzed by superoxide dismutase to form hydrogen peroxide, the hydrogen peroxide can be catalyzed by iron ions to form hydroxyl radicals, macromolecular damages such as DNA and protein are generated, and the structures of mitochondria and cell membranes are damaged, and serious pathological aging is caused.

Normally, the level of pro-oxidant and the level of antioxidant in the body are in dynamic balance, but the generation and the existence of ROS are regional, and the important difference between a polar phase and a non-polar phase exists, and the elimination of ROS also has two modes of other metabolic free energy driving and reaction self free energy driving, so that in cell aging metabolic disorder, mitochondrial damage causes insufficient free energy supply, and pathological phenomena such as ROS local runaway and the like are often generated. The high-concentration ROS damages biomacromolecules in cells, inhibits an organism antioxidant system, reduces activities of antioxidant enzymes such as superoxide dismutase, catalase and glutathione-S-transferase, further aggravates imbalance of the level of an oxidant and an antioxidant, and accelerates the pathological aging process of a female reproductive system.

DNA is an important target for ROS attack, ROS can cause DNA strand breaks, site mutations, double strand aberrations leading to DNA deamination, base oxidation, etc., and mitochondrial DNA is more susceptible to oxidative damage than nuclear DNA. Most forms of DNA mutation by ROS oxidize G to 8-oxoguanine (8-oxoG), and 8-oxoG no longer pairs with C but pairs with A, so that G is converted into A, and the generation of progressive DNA damage causes the continuous accumulation of gene mutation, which is an important reason for pathological aging.

Nicotinamide adenine dinucleotide phosphate oxidase (NADPH oxidase, Nox) reduces oxygen molecules in vivo into superoxide anions through an NADPH dependent process, is the only enzyme which directly generates ROS in vivo, has a signal transduction effect in cells by catalyzing the generated moderate ROS, and is an essential regulatory factor for processes such as cell proliferation and the like. And Nox production of ROS is a free energy driven, controlled metabolic process that is derived from NADPH produced by cellular metabolism and has some complementarity to mitochondrial leakage of ROS.

In conclusion, the core of female reproductive system aging is cellular aging, wherein DNA damage, oxidative stress and mitochondrial dysfunction are major causes of pathological cellular aging, and intracellular ROS metabolic disorder is the root cause of DNA damage, oxidative stress and mitochondrial dysfunction, so that pathological damage induced by the ROS factors other than the telomere-dominated physiological aging is called ROS metabolic disorder-related aging damage.

At present, the problems of great side effect or poor effect and the like generally exist in a prevention health care scheme for relieving cell ROS aging injury, for example, patents with application numbers of 201810104899.X, 201610041188.3, 201910680707.4 and the like try to utilize probiotics and the like to effectively maintain flora balance, the treatment effect of probiotic combination used for regulating intestinal flora in the prior art is not doubtful, more importantly, the flora of female reproductive systems such as vagina and the like is obviously different from the intestinal tract, and the intestinal probiotic combination is not suitable for being settled in tissues such as the vagina and the like, so the technical route has fundamental defects and good treatment effect. The traditional Chinese medicine complexing agent is used for regularly cleaning the vagina in the patents with application numbers of 201811644710.2, 201910893065.6 and the like, firstly, the traditional Chinese medicine formula is complex, the traditional Chinese medicine complexing agent contains various complex chemical components such as alkaloid and the like, the problem of causing allergy of patients and the like is difficult to ensure, in addition, the cleaning may damage the lactic acid bacteriostatic environment formed by vaginal glycogen fermentation, the vagina resistance is weakened, and additional infection is caused. Patent 201710140768.2 utilizes vitamin E, folic acid, vitamin B12, etc. to promote reproductive health in women of childbearing age, and such nutrients are not deficient in the dietary intake of most women, and excessive supplementation may not be beneficial or harmful. In conclusion, the female reproductive system is a complex multi-organ interaction system, so that the problem can be solved only by starting from the problem of cell aging, but no safe and effective mature scheme aiming at preventing and treating the pathological aging of the female reproductive system by cell ROS damage exists until the application of a patent.

Disclosure of Invention

The general formula of Nicotinic acid derivatives (Nicotinic derivatives) is shown in FIG. 1, wherein R is structure A or structure B₁And R₂Can be a hydrogen atom, an amine group or other group (e.g., nicotinamide is R in structure B₁Is an amino group, nicotinamide mononucleotide is R in structure A₁Is amino, R₂Is a nucleotide group), X^-Is a certain electronegative group.

Intracellular ROS are produced in the metabolism and eliminated in the metabolism as the key of the pathological female reproductive system aging damage, the root of the ROS metabolic disturbance is a metabolic problem, and the core of metabolic regulation is various high-efficiency catalytic enzymes and cofactors, so that the correction of the ROS metabolic disturbance should be started from the enzymes and the cofactors thereof.

Specifically, along with the physiological or pathological aging process of the female reproductive system, the intracellular pro-oxidant level and the antioxidant level begin to be unbalanced, and especially, the decrease of the activities of antioxidase such as superoxide dismutase, catalase, glutathione S-transferase and the like causes the increase of ROS concentration and aggravation of damage, so that a small molecular substance for gently promoting the expression level of the antioxidase needs to be searched.

Starting from the theory of biochemistry and molecular biology, in the human cofactor for eliminating ROS enzymes, Nicotinamide Adenine Dinucleotide (NADH) and Nicotinamide Adenine Dinucleotide Phosphate (NADPH) are in the core position, NADH exists in each cell, and the NADH directly participates in a plurality of physiological activities of substance metabolism, energy synthesis, cell DNA repair and the like of human vital activities. Of these, NADH is most noteworthy the only substrate for DNA repair enzymes and is an important raw material for intracellular DNA repair systems. DNA damage is a major root cause of cell aging and the occurrence of various diseases; and NADH can also directly improve the functions of mitochondria, prevent ROS leakage of the mitochondrial respiratory chain and reduce the ROS yield from the source. More importantly, NADH is a core cofactor for eliminating ROS in the body, is used as a transfer electron, is a coenzyme of a plurality of dehydrogenases in the body, for example, NADH can transfer reduced free energy to GSH through NADPH to eliminate lipid peroxidation and the like, so that NADH is a core intermediate carrier for actively eliminating the free energy and the reduction potential of ROS by the body, and is a key for delaying the pathological aging damage of the female reproductive system caused by ROS.

However, NADH and DNADP are too large in molecular weight to be directly absorbed by the human body. In view of the above, the inventor of the present invention has found through long-term experimental screening and verification that despite the differences in effective dosage and action effect, nicotinic acid derivatives such as nicotinamide, nicotinic acid inositol ester, nicotinamide riboside, nicotinic acid riboside and nicotinamide mononucleotide can all improve the ability of female reproductive system cells to resist pathological oxidative damage, because from the biochemical metabolism, they can all be used as effective sources for synthesizing nicotinyl groups of NADH and DNADP by cells, but there are differences in pharmacokinetics.

On the basis of the analysis and the experiment, the inventor refers to a theoretical framework that the core of female reproductive system aging damage is cell aging, wherein the core of female reproductive system aging damage is systematically clarified in a previous working research stage is cell aging, and DNA damage, oxidative stress and mitochondrial dysfunction induced by ROS metabolic disorder are main reasons for generating pathological cell aging damage, and specifically screens an ROS metabolic disorder correction formula under the guidance of the theoretical framework, and then the conclusion that the nicotinic acid-based derivative is most suitable for preventing and treating the ROS-induced cell damage is obtained through long-term cell experiments and random double-blind control experiments of human volunteers through improvement and verification.

Compared with the prior art, the invention has the following advantages.

1. The invention has strong innovation, the inventor gradually establishes a theoretical framework that 'the core of female reproductive system aging is cell aging, wherein DNA (deoxyribonucleic acid) damage, oxidative stress and mitochondrial dysfunction induced by ROS (reactive oxygen species) metabolic disturbance are main reasons for generating pathological aging damage of cells' on the basis of long-term research on ROS (reactive oxygen species) damage cell mechanism of the female reproductive system, and under the guidance of the theoretical framework, a mature technical scheme for comprehensively relieving the ROS of the cells is pertinently screened, and the expression level of antioxidant enzyme is improved by adding nicotinic acid-based derivatives, so that the ROS-induced cell damage is relieved.

About 3% of oxygen intake in the body leaks in the respiratory chain of mitochondria and is converted into ROS, and the actual mechanism of eliminating ROS by using a reducing agent such as dihydrolipoic acid is to use free energy and reduction potential generated by metabolism of the body as a switch, but the free energy can only come from the metabolic process of a pentose phosphate pathway, so that the simple supplement effect is poor. The inventor utilizes nicotinic acid-base derivative to promote the generation of NADH and NADPH, which are important hydrogen carriers of organisms and are also main carriers of free energy and reduction potential generated by metabolism, and can continuously reduce the oxidized lipoic acid into the dihydrolipoic acid, thereby reliably exerting the effect of each other and eliminating the ROS level of cells.

The body has free-energy driven ROS elimination and also non-free-energy driven ROS elimination, and nicotinic acid-based derivatives can improve the activity level of the ROS eliminated free-radical driven enzyme system. However, the free energy and non-free energy systems for eliminating ROS are interdependent and complementary, for example, superoxide anion leaked by mitochondria must be converted into hydrogen peroxide by superoxide dismutase, the latter is converted into hydroxyl radical to damage a cell membrane system by fenton reaction catalyzed by iron ions, and then the hydrogen peroxide must be eliminated by GPX4 driven by free energy, while the free energy of GPX4 comes from a metabolic pathway of NADPH-GSH, therefore, the supplementation of nicotinic acid-based derivatives can play an important auxiliary role of increasing NADPH concentration and the like, and the ROS damage of cells can be effectively eliminated.

More importantly, cellular physiology requires certain concentrations of ROS levels as secondary messengers for proliferation, but current protocols fail to address the problem of excessive interference of ROS elimination with cellular metabolism. However, the concentration of NADPH is increased by supplementing the tobacco base derivative, and ROS is generated by the catalytic reaction of NADPH oxidase, so that the high concentration of NADPH can promote the generation of ROS which is believed to be used in a specific cell region, and further maintain and promote the cell activity.

2. The formula of the invention has obvious effect from the experimental data of a cell nucleus animal model.

See examples 1 and 2 for details, the compound formula has obvious effect on standardized high-flux cell and animal model experiments.

3. From the trial data of volunteers, the formula of the invention is obviously stronger than the mainstream scheme in the market.

In terms of action and effect, the cell viability experiment and experimental methods such as random double-blind comparison of a large sample of a plurality of centers of volunteers are comprehensively utilized, statistical analysis is carried out on the basis of obtaining a large amount of data, and the effect of the formula is found to be better than that of certain marketed formulas (see example 3 in detail).

4. In terms of safety, the ingredients of the present protocol are derived from metabolism-regulatory nutrients recognized in the industry as having low toxicity, and the cytotoxicity of the formulation was specifically checked in preliminary experiments, ensuring that the dose used did not exceed 10% of the toxicity threshold. In addition, the early work of the invention accurately proves the action range and the toxicity threshold of the components through a standardized high-flux cell experiment, thereby ensuring that the use dosage of each component has certain tolerance and being beneficial to the development and marketing of products.

Drawings

FIG. 1 shows the general structure (structure A or structure B, X) of Nicotinic acid derivatives (Nicotinic derivatives)^-Is a certain electronegative group, wherein R₁And R₂Can be a hydrogen atom, an amine group or other group, e.g. nicotinamide is R in structure B₁Is an amino group, nicotinamide mononucleotide is R in structure A₁Is amino, R₂Is a nucleotide group).

Detailed description of the preferred embodiments.

The technical embodiments of the present invention will be described in further detail with reference to specific examples.