Application of mannose sodium in preparation of anti-aging medicine

Abstract

The invention discloses an application of mannose sodium in preparation of an anti-aging drug. Researches show that the mannose sodium has a relatively good anti-aging effect, can improve appetite and memory of an aging model animal, can remarkably reverse organ atrophy caused by D-galactose, can up-regulate liver antioxidant enzyme SOD (superoxide dismutase) and CAT (catalase) level activity, can down-regulate the content of MDA (malondialdehyde) in the liver, can relieve oxidative stress and can delay aging; meanwhile, the mannose sodium can reduce the activity of beta-galactosidase (SA-beta-gal) of aging animals, has the effect of inhibiting SA-beta-gal, can delay fat aging of aging mice induced by high fat diet, and provides more effective drugs for research and development of novel anti-aging drugs. And the mannose sodium has wide pharmacological activity, remarkable treatment effect, small toxic and side effects and high safety, and can replace beta-nicotinamide mononucleotide to be used as a new anti-aging drug for delaying aging.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and more specifically, relates to the application of sodium mannitol in the preparation of anti-aging drugs. Background Art

[0002] Aging is a degenerative disease in which the body's functions deteriorate and the functional decline of tissues and organs occurs over time. The free radical aging theory believes that the increase in oxidative stress response is an important driving factor for aging, and the antioxidant capacity comes from superoxide dismutase (SOD), which exists in every body. Oxidative stress (OS) refers to a state of imbalance between oxidation and antioxidant effects in the body, which tends to oxidize, leading to inflammatory infiltration of neutrophils, increased secretion of proteases, and the production of a large number of oxidative intermediates. Oxidative stress is a negative effect produced by free radicals in the body and is considered to be an important factor leading to aging and disease. With the increase in life expectancy, the activity of SOD in the human body will gradually decrease until the free radicals cannot be effectively removed and accumulate in the body, leading to aggravated aging. With the progress of aging, the incidence and prevalence of diseases such as coronary heart disease, diabetes, and atherosclerosis will increase. The harm of excessive oxidation in the human body will accelerate aging, disease, and death; cause rapid growth of tumors; cause inflammation and autoimmune reactions that damage physical health; produce pigmentation and spots. Increasing the superoxide dismutase (SOD) content of the body to avoid oxidation plays an important role in resisting the aging of the body. Therefore, it is of great significance to find more anti-aging drugs that can effectively delay aging and prevent and treat aging and related diseases.

[0003] Chemical drug treatment is an important means of anti-aging at present. At present, the anti-aging drug used in clinical practice to delay aging is mainly β-nicotinamide mononucleotide (NMN), and existing studies have reported the anti-aging effect of β-nicotinamide mononucleotide. However, the application of β-nicotinamide mononucleotide also has many limitations, and the side effects of this product are a major problem in anti-aging work. The most common side effects reported in clinical trials of β-nicotinamide mononucleotide are hypoglycemia, insomnia, and acne. At the same time, long-term, large-scale and repeated use of β-nicotinamide mononucleotide can easily cause the body to develop tolerance to it, resulting in a decrease in its anti-aging effect. Therefore, it is urgent to find an anti-aging drug that is highly safe, has fewer side effects, has good anti-aging effects, and can replace β-nicotinamide mononucleotide. It is of great significance and practical application value to provide more anti-aging drugs. Summary of the invention

[0004] The technical problem to be solved by the present invention is to overcome the defects and shortcomings of existing anti-aging drugs and provide a new application of sodium mannitol in the preparation of anti-aging drugs.

[0005] The purpose of the present invention is to provide the application of sodium mannitol in preparing anti-aging medicine.

[0006] Another object of the present invention is to provide the use of sodium mannitol in the preparation of β-galactosidase activity inhibitors.

[0007] The above-mentioned purpose of the present invention is achieved through the following technical solutions:

[0008] The present invention provides a new application of the old drug mannitol sodium. By establishing an aging animal model induced by D-galactose and a fat aging model for in vivo experiments, the anti-aging effect of mannitol sodium is confirmed. Studies have shown that mannitol sodium can improve the appetite and memory of aging model animals, increase appetite, significantly increase food intake, and effectively improve the memory of aging mice; increase the spleen organ index of aging mice, significantly reverse the organ atrophy caused by D-galactose, upregulate the activity of liver antioxidant enzymes superoxide dismutase (SOD) and catalase (CAT), downregulate the content of malondialdehyde (MDA) in the liver, relieve oxidative stress, and delay aging. At the same time, mannitol sodium can reduce the activity of β-galactosidase (SA-β-gal) in aging animals, has the effect of inhibiting SA-β-gal, can delay the fat aging of aging mice induced by a high-fat diet, and provide more effective drugs for the development of new anti-aging drugs. In addition, sodium mannitol has a wide range of pharmacological activities, with little toxicity and side effects, high safety, and significant effects. It can replace the anti-aging drug β-nicotinamide mononucleotide in delaying aging, and has important significance and practical application value in providing more anti-aging drugs.

[0009] Therefore, the present invention provides the use of sodium mannitol in preparing anti-aging medicines.

[0010] Furthermore, the drug can increase appetite.

[0011] Furthermore, the drug can improve aging-related memory ability.

[0012] Furthermore, the drug can delay the atrophy of aging organs.

[0013] Preferably, the method is capable of reversing atrophy of the aged spleen organ.

[0014] Further, the drug can alleviate oxidative stress.

[0015] Furthermore, the drug can increase the contents of liver SOD and liver CAT.

[0016] Furthermore, the drug can reduce the MDA content in the liver.

[0017] Further preferably, the drug can delay fat aging.

[0018] The sodium mannitol (GV-971) used in the present invention is a low molecular weight acidic oligosaccharide compound, i.e., oligosaccharide, prepared from marine brown algae extract, which has a wide range of pharmacological activities and is a new drug for the treatment of Alzheimer's disease that targets the brain-gut axis. It has good in vivo safety and few toxic and side effects. The structural formula of sodium mannitol is shown in the following formula (I):

[0019]

[0020] The invention also provides application of mannitol sodium in preparing beta-galactosidase activity inhibitor.

[0021] Preferably, the dosage form of the drug is capsule, tablet, oral preparation, microcapsule preparation or injection.

[0022] The present invention has the following beneficial effects:

[0023] The invention provides a new application of sodium mannitol in the preparation of a drug for delaying aging. Studies have shown that sodium mannitol has a good anti-aging effect, can improve the appetite and memory of aging model animals, significantly reverse organ atrophy caused by D-galactose, upregulate the activity of liver antioxidant enzymes SOD and CAT levels, downregulate the content of MDA in the liver, relieve oxidative stress, and delay aging; at the same time, sodium mannitol can reduce the activity of β-galactosidase (SA-β-gal) of aging animals, has the effect of inhibiting SA-β-gal, can delay fat aging of aging mice induced by a high-fat diet, and provides more effective drugs for the research and development of new anti-aging drugs.

[0024] The anti-aging drug sodium mannitol provided by the present invention has a wide range of pharmacological activities, including anti-neuroinflammatory and anti-tumor effects, and has small toxic and side effects. No toxic effects or deaths were found in mice given 400 mg / kg of sodium mannitol by gavage. It has better safety and less toxic and side effects than β-nicotinamide mononucleotide, and can replace the use of β-nicotinamide mononucleotide in delaying aging, providing more possibilities and treatment means for avoiding clinical tolerance. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 This is a statistical chart of the food intake results of aging mice after the action of sodium mannitol.

[0026] Figure 2 This is a statistical chart of the Y-maze results of aging mice after the treatment of sodium mannitol.

[0027] Figure 3 This is a statistical chart of the spleen organ index of aging mice after the action of sodium mannitol.

[0028] Figure 4This is a statistical chart of T-SOD, CAT and MDA levels in the liver of aging mice after the treatment of sodium mannitol.

[0029] Figure 5 This is the staining result of SA-β-gal in the inguinal fat of aged mice after the action of sodium mannitol.

[0030] Figure 6 This is a statistical chart of SA-β-gal levels in the inguinal fat of aged mice after the action of mannitol sodium.

[0031] The quantitative results in the figures are expressed as mean ± standard error (SEM). All data were statistically analyzed using GraphPad Prism 9.0 (GraphPad Software, San Diego, USA). The differences among the groups were analyzed by one-way analysis of variance (ANOVA). All bar graphs were drawn by GraphPad Prism 9.0. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. DETAILED DESCRIPTION

[0032] The present invention is further described below in conjunction with the accompanying drawings and specific examples, but the examples do not limit the present invention in any form. Unless otherwise specified, the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in the art.

[0033] Unless otherwise specified, the reagents and materials used in the following examples are commercially available.

[0034] The experimental animals C57BL / 6J mice used in the following examples were purchased from Beijing Weitonglihua Experimental Animal Co., Ltd., D-galactose was purchased from Shanghai MacLean Biochemical Technology Co., Ltd.; and mannitol sodium (GV-971) was purchased from Shanghai Green Valley Pharmaceutical Co., Ltd.

[0035] Example 1 Establishment of an aging mouse model

[0036] 1. D-galactose-induced aging model

[0037] Eight-week-old male C57BL / 6J mice were randomly divided into a normal control group (control, n = 9), a D-galactose modeling group (aging, n = 11) and a treatment group (n = 9). Except for the normal control group, the remaining mice were subcutaneously injected with D-galactose (1000 mg / kg / day) for 63 consecutive days, while the normal control group (control) was subcutaneously injected with the same volume of saline (10 μL / g).

[0038] D-galactose modeling group (aging): mice were subcutaneously injected with D-galactose (1000 mg / kg / day).

[0039] The treatment groups were treated with sodium mannitol prepared with 0.9% saline. Mice were gavaged with 200 mg / kg and 400 mg / kg sodium mannitol every day, respectively, which were recorded as 200 mg / kg sodium mannitol treatment group (LG+aging) and 400 mg / kg sodium mannitol treatment group (HG+aging).

[0040] The treatment group was gavaged with sodium mannitol, and the D-galactose modeling group (aging) and the normal control group (control) were gavaged with the same volume of normal saline, once a day for a total of 63 days.

[0041] 2. Fat aging mouse model

[0042] Since obesity accelerates the aging process of the human body, an obesity model was constructed by inducing a high-fat diet, and aging analysis was performed in combination with the activity of the adipose aging marker β-galactosidase (SA-β-gal). The adipose aging mouse model was successfully induced by feeding mice with a high-fat diet (60kcal% fat) for 10 weeks.

[0043] A normal control group and a treatment group were set up at the same time, with 5 mice in each group, and the entire experimental period lasted for 26 weeks. The treatment group was first given a high-fat diet for 10 weeks to successfully induce an obese mouse model, and then given sodium mannitol for intervention treatment and maintained a high-fat diet. Sodium mannitol was used for intervention at two doses of 200 mg / kg and 400 mg / kg. Sodium mannitol was used for a total of 15 weeks of intervention, and samples of mice in each group were collected after the end of the experimental period.

[0044] Example 2 Anti-aging effect of sodium mannitol

[0045] For the D-galactose-induced aging model, the food intake of mice was tested daily. On the 61st day after subcutaneous injection of D-galactose, the Y-maze behavioral test of mice was performed. The specific method refers to the prior art: Xiao Y, et al. Human gut-derived B. longum subsp. longum strains protect against aging in a d-galactose-induced aging mouse model. Microbiome 9, 1-21 (2021). On the 64th day after subcutaneous injection of D-galactose, the weight of mice was measured, and then the mice were anesthetized with isoflurane gas and blood was collected from the eye sockets. The blood was left to stand at room temperature for more than one hour, centrifuged at 5000 rpm and 4°C for 20 minutes, and then the serum was collected. Then the mice in different treatment groups were taken, cervical dislocation was performed, and the spleen and liver were collected. The spleen organ index (ratio of spleen weight to body weight) was detected, and the indicators of liver oxidative stress phenotype were detected using kits: liver SOD (kit model A001-1-2, Nanjing Jiancheng), liver CAT (kit model A007-1-1, Nanjing Jiancheng) and liver MDA (kit model A003-1-2, Nanjing Jiancheng).

[0046] The statistical results of food intake of aged mice are as follows Figure 1 As shown in the figure, compared with the normal control group, the daily food intake of mice in the aging group was significantly reduced, indicating that the appetite of aging mice decreased; after the intervention of sodium mannitol, the daily food intake of mice in the sodium mannitol intervention group was significantly increased, and the difference was statistically significant, indicating that sodium mannitol can effectively improve the appetite of aging mice.

[0047] The behavioral test results of aging mice are as follows Figure 2 As shown, compared with the normal control group, the spontaneous round-robin behavior scores of mice in the aging group decreased, indicating that the memory of aging mice declined; after the intervention of sodium mannitol, the spontaneous round-robin behavior scores of mice in the sodium mannitol intervention group increased significantly, and the difference was statistically significant, indicating that sodium mannitol can effectively improve the memory of aging mice.

[0048] The statistical results of spleen organ index of aged mice are as follows Figure 3 As shown, compared with the normal control group, the spleen organ index of the aging group mice decreased, indicating that the spleen organ of the aging mice was atrophied; after the intervention of sodium mannitol, the spleen organ index of the mice in the sodium mannitol intervention group increased, and the difference was statistically significant, indicating that sodium mannitol can significantly reverse the atrophy of the spleen organ in aging mice.

[0049] The statistical results of liver biomarkers of aging in each group of mice are as follows Figure 4 As shown in the figure, compared with the aging group, the SOD ( Figure 4 A) CAT( Figure 4 B) levels increased significantly, liver MDA ( Figure 4 C) levels decreased significantly, and the difference was statistically significant, indicating that sodium mannitol has a good anti-aging effect and can effectively improve the levels of liver biomarkers in aging mice.

[0050] Example 3 Inhibitory activity of sodium mannitol on β-galactosidase

[0051] For the aging mouse model induced by a high-fat diet, at the 26th week of high-fat diet, the mice were anesthetized with isoflurane gas and then dissected to obtain inguinal fat. Inguinal fat samples were taken from mice in each treatment group, and the activity of β-galactosidase (SA-β-gal), a marker of mouse inguinal fat aging, was detected using a kit (CBA-230, CELL BIOLABS).

[0052] The staining results of the aging marker SA-β-gal are shown in the figure Figure 5 As shown in the figure, compared with the normal control group, the blue area ratio of the inguinal fat in the aging group increased significantly, indicating that the SA-β-gal activity of the inguinal fat of the aging mice increased significantly; compared with the aging group, the blue area ratio of the inguinal fat in the mannitol sodium intervention group decreased significantly, indicating that the SA-β-gal activity of the inguinal fat of the aging mice decreased significantly. The statistical chart of the aging marker SA-β-gal is shown in the figure below. Figure 6 As shown, compared with the normal control group, the SA-β-gal activity of the inguinal fat in the aging group was significantly increased, indicating that the inguinal fat of the aging mice was aged; compared with the aging group, the SA-β-gal activity of the inguinal fat in the mannitol sodium intervention group was significantly decreased, and the difference was statistically significant, indicating that mannitol sodium can inhibit the activity of β-galactosidase, has anti-aging effects, and can effectively improve the aging of the inguinal fat of aging mice.

[0053] In summary, it is shown that mannitol sodium has a good anti-aging effect, can improve the appetite and memory of aging mice induced by D-galactose, significantly reverse the organ atrophy caused by D-galactose, upregulate the activity of liver antioxidant enzymes, and downregulate the content of MDA in the liver to relieve oxidative stress; at the same time, it can reduce the activity of β-galactosidase in the inguinal fat of aging mice induced by a high-fat diet, and delay aging. Therefore, the present invention provides a new use of mannitol sodium in anti-aging. Since mannitol sodium has small toxic and side effects and high in vivo safety, it can replace the use of the anti-aging drug β-nicotinamide mononucleotide, and at the same time provide more safe and effective anti-aging drugs, which is of great significance and practical application value for providing more anti-aging drugs.

[0054] The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be equivalent replacement methods and are included in the protection scope of the present invention.

Claims

1. Application of sodium mannitol in the preparation of anti-aging drugs.

2. The use according to claim 1, characterized in that: The drug can increase appetite.

3. The application according to claim 2, characterized in that: The drug can improve aging-related memory ability.

4. The use according to claim 3, characterized in that: The medicine can delay the atrophy of aging organs.

5. The application according to claim 4, characterized in that: The drug can alleviate oxidative stress.

6. The use according to claim 5, characterized in that: The drug can increase the levels of superoxide dismutase and catalase in the liver.

7. The use according to claim 5, characterized in that: The drug can reduce the content of malondialdehyde in the liver.

8. The use according to any one of claims 1 to 7, characterized in that: The medicine can delay fat aging.

9. Application of sodium mannitol in the preparation of β-galactosidase activity inhibitors.

10. The use according to claim 1 or 9, characterized in that: The dosage form of the drug is capsule, tablet, oral preparation, microcapsule preparation or injection.

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