Use of organometallic macrocyclic compound molecule in catalytic enhancement of level of intracellular oxidized coenzyme i and in Anti-aging product

Abstract

Disclosed in the present invention is use of an organometallic macrocyclic compound molecule in the catalytic enhancement of the level of intracellular oxidized coenzyme I and in an anti-aging product. The organometallic macrocyclic compound molecule of the present invention enters a cell and catalyzes the conversion of reduced coenzyme I within the cell into oxidized coenzyme I, thereby enhancing the metabolic activity of the cell and prolonging the lifespan of aging animals. The organometallic macrocyclic compound molecule is applied to oxidized coenzyme I supplements, anti-aging drugs, and health care products. By means of daily oral administration, the organometallic macrocyclic compound molecule can improve the metabolic activity of the aging population, thereby maintaining their physiological health and prolonging their lifespan.

Description

Application of an organometallic macrocyclic compound molecule in catalytic enhancement of intracellular oxidized coenzyme I levels and in anti-aging products Technical Field

[0001] This invention relates to the field of biomedicine, specifically to the application of an organometallic macrocyclic compound molecule in catalyzing the enhancement of intracellular oxidized coenzyme I levels and in anti-aging products. Background Technology

[0002] Nicotinamide adenine dinucleotide (coenzyme I) is an important coenzyme in cells, involved in energy metabolism and redox reactions. With age, the level of oxidized coenzyme I in the body gradually declines, and this reduction is considered one of the key factors in the aging process. As the level of oxidized coenzyme I decreases, many cellular functions are affected, including DNA repair, mitochondrial energy production, immune system response, cell viability maintenance, and epigenetic regulation. The weakening of these functions leads to the accumulation of DNA damage, reduced mitochondrial activity, decreased immunity, and the senescence of cells, which in turn affects the regenerative capacity of tissues, causing organs to gradually degenerate. Therefore, increasing or maintaining the level of oxidized coenzyme I can help slow down the aging process and, to some extent, reduce age-related health risks.

[0003] There are two main pathways for supplementing oxidized coenzyme I: the de novo synthesis (DNS) pathway and the rescue pathway. The DNS pathway involves seven enzymatic steps to synthesize oxidized coenzyme I from the substrate tryptophan, a process that is extremely inefficient within cells. In contrast, the rescue pathway utilizes vitamin B3 derivatives, such as nicotinamide (NAM), nicotinamide mononucleotide (NMN), and nicotinamide nucleoside (NR), to bypass the intracellular adenine synthesis process and synthesize oxidized coenzyme I in vivo with fewer steps. Studies have found that supplementing oxidized coenzyme I with tryptophan can effectively improve the health of fruit flies and increase their lifespan (Aging Cell, 2024, 23(4):e14102.). In the rescue pathway, feeding NAM to nematodes, mice, and humans can increase the level of oxidized coenzyme I in these organisms, thereby improving their age-related muscle aging and degenerative muscle diseases (Cell reports, 2021, 34(3)). The use of NMN and NR has been shown to increase the level of oxidized coenzyme I in aging mice and prolong their lifespan (Cell metabolism, 2016, 24(6):795-806; Science, 2016, 352(6292):1436-1443). However, recent studies have found that supplementing with precursor substances can also increase the level of reduced coenzyme I, and excessive reduced coenzyme I can lead to related diseases (Nature medicine, 2024, 30(2):424-434).

[0004] In summary, both de novo synthesis and salvage pathways still require multiple steps of conversion into oxidized coenzyme I within the cell. Precursor supplementation can significantly increase the level of oxidized coenzyme I in the body, but it will also lead to the accumulation of reduced coenzyme I in the body. High levels of reduced coenzyme I can cause a series of diseases in the body (such as cardiovascular diseases, skin diseases, tumors, etc.). Summary of the Invention

[0005] This invention addresses the shortcomings of existing technologies by providing an application of organometallic macrocyclic compounds in catalyzing the enhancement of intracellular oxidized coenzyme I levels and in anti-aging products. By allowing organometallic macrocyclic compounds to enter cells and catalyze the conversion of reduced coenzyme I to oxidized coenzyme I, the level of oxidized coenzyme I is efficiently increased through a one-step catalytic method while simultaneously reducing the level of reduced coenzyme I. This improves cellular metabolic activity, combats aging, and ultimately extends the lifespan of aging animals, maintaining their physiological health.

[0006] To address the aforementioned technical problems, this invention provides the application of organometallic macrocyclic compounds in catalytically enhancing intracellular oxidized coenzyme I levels and in anti-aging products.

[0007] Furthermore, the structural formula of the organometallic macrocyclic compound molecule is as follows:

[0008] Where M is a metal, R1-R 12 Independently selected from chain alkyl, cyclic alkyl, substituted alkyl, hydrogen or aromatic groups, R 13 Selected from hydrogen, hydroxyl, halogen groups, or oxygen. Organometallic macrocyclic compounds may also contain macrocyclic structures with more than 13 members.

[0009] Furthermore, M is Cu, Fe, Co, Ni, Mn, Zn, Pt, Pb, V, Ag, Sn, Gd, Ga, Cd, Ir, Rh, or In.

[0010] Furthermore, the catalytic enhancement of intracellular oxidized coenzyme I levels specifically involves: catalytically converting intracellular reduced coenzyme I into oxidized coenzyme I, thereby increasing intracellular oxidized coenzyme I levels.

[0011] Furthermore, the cells include cardiomyocytes, hepatocytes, lung cells, fibroblasts, epithelial cells, and immune cells.

[0012] Furthermore, the immune cells include macrophages, T cells, NK cells, and B cells.

[0013] Furthermore, the organometallic macrocyclic compound molecule improves mitochondrial metabolic function by increasing intracellular levels of oxidized coenzyme I.

[0014] Furthermore, the functions of the mitochondria include the ability to synthesize ATP and the ability to perform aerobic respiration.

[0015] Furthermore, the product includes oxidized coenzyme I supplements.

[0016] Furthermore, the products include anti-aging medicines or health supplements.

[0017] The beneficial effects of this invention are:

[0018] The organometallic macrocyclic compound molecules of this invention enter the cell and catalyze the conversion of reduced coenzyme I into oxidized coenzyme I. While reducing the level of reduced coenzyme I, it efficiently increases the level of oxidized coenzyme I in the cell through a one-step catalytic method, thereby improving cell activity, anti-aging, and thus prolonging the lifespan of aging animals and maintaining their physiological health.

[0019] The organometallic macrocyclic compound molecules of this invention can be applied to oxidized coenzyme I supplements, anti-aging drugs or health products. Daily use can increase the lifespan of aging people and greatly improve their physiological health. Attached Figure Description

[0020] To more clearly illustrate the technical solution of the present invention, the drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0021] Figure 1 is a comparison of the drug administration concentration of the organometallic macrocyclic compound in Example 1 of the present invention with the ratio of oxidized coenzyme I to reduced coenzyme I in cells.

[0022] Figure 2 is a comparison of mitochondrial activity in cells before and after administration of the organometallic macrocyclic compound molecule in Example 2 of the present invention.

[0023] Figure 3 is a comparison of the ratio of oxidized coenzyme I to reduced coenzyme I in mouse liver before and after administration of the organometallic macrocyclic compound molecule in Example 3 of the present invention.

[0024] Figure 4 is a comparison of the lifespan extension of mice before and after administration of the organometallic macrocyclic compound molecule in Example 4 of the present invention.

[0025] Figure 5 is a comparison of the improvement in mouse health before and after administration of the organometallic macrocyclic compound molecule in Example 5 of the present invention. Detailed Implementation

[0026] The technical solution of the present invention will be clearly and completely described below with reference to specific embodiments. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0027] This embodiment provides an application of an organometallic macrocyclic compound molecule in catalytically enhancing intracellular oxidized coenzyme I levels and in anti-aging products. The structural formula of the organometallic macrocyclic compound molecule is as follows:

[0028] Where M is a metal, R1-R 12 Independently selected from chain alkyl, cyclic alkyl, substituted alkyl, hydrogen or aromatic groups, R 13 It is selected from hydrogen, hydroxyl, halogen groups or oxygen.

[0029] As a specific example, M is Cu, Fe, Co, Ni, Mn, Zn, Pt, Pb, V, Ag, Sn, Gd, Ga, Cd, Ir, Rh, or In.

[0030] As a specific example, the catalytic enhancement of intracellular oxidized coenzyme I levels specifically involves: catalyzing the conversion of intracellular reduced coenzyme I into oxidized coenzyme I, thereby increasing intracellular oxidized coenzyme I levels; the cells include cardiomyocytes, hepatocytes, lung cells, fibroblasts, epithelial cells, and immune cells; the immune cells include macrophages, T cells, NK cells, and B cells.

[0031] Specifically, the organometallic macrocyclic compound molecule improves mitochondrial function by increasing the level of oxidized coenzyme I in cells, and the mitochondrial function includes ATP synthesis capacity and aerobic respiration capacity.

[0032] As a specific example, the products include oxidized coenzyme I supplements, anti-aging drugs, or health products.

[0033] The following are specific examples.

[0034] Example 1

[0035] This embodiment investigates the effect of organometallic macrocyclic compounds on increasing intracellular oxidized coenzyme I levels. HepG2 hepatocytes were incubated with different concentrations of the organometallic macrocyclic compound hemin, and the intracellular reduced coenzyme I and oxidized coenzyme I levels were detected by LC-MS / MS. The ratio of oxidized coenzyme I to reduced coenzyme I in each cell was calculated.

[0036] The extraction methods for intracellular reduced coenzyme I and oxidized coenzyme I were as follows: After processing, cells in a 6-well plate were cleaned and washed three times with pre-chilled PBS (4°C). 1 mL of phosphate-buffered saline (PBS) was added to each well, and cells were scraped from the bottom of the plate into the PBS using a cell scraper. The cells were thoroughly mixed with a pipette and transferred to a 1.5 mL centrifuge tube. The centrifuge tube containing the cell suspension was centrifuged at 1000 rpm for 5 min, and the supernatant was removed. 50 μL of pre-chilled deionized water (4°C) was added, and the cell pellet was resuspended and subjected to repeated freeze-thaw cycles in liquid nitrogen (3 cycles, 10 min each). 50 μL of the lysate was mixed with 200 μL of pre-chilled methanol and incubated at -80°C for 20 min, followed by centrifugation at 20000 g for 10 min at 4°C. The supernatant was collected to obtain the cell lysate. Add 150 μL of methanol extraction buffer (-20℃) to the cell pellet, and pipette to mix the cell pellet thoroughly. Centrifuge at 20000g for 10 min, collect the supernatant, and mix it with the previous extraction buffer. Dry the extraction buffer with nitrogen gas and store it at -80℃ for later analysis.

[0037] Detection method:

[0038] a) Reduced Coenzyme I and Oxidized Coenzyme I: The sample injection volume was set to 5 μL. A C18 reversed-phase column (2.1 mm × 150 mm) was used, with an elution flow rate of 300 μL / min. Mobile phase A: 20 mM ammonium acetate solution; mobile phase B: 100% acetonitrile. The mobile phase gradient for sample separation was as follows: 98% mobile phase A was used for isocratic separation for 5 min, followed by a linear change of 10% gradient in mobile phase A within 2 min, then isocratic separation was maintained at 10% mobile phase A for 2 min, followed by a linear change of 98% gradient in mobile phase A within 0.1 min.

[0039] b) Dihydroxyacetone phosphate: The injection volume, column and flow rate were the same as those of reduced coenzyme I. The separation gradient was set as follows: 95% mobile phase A isocratic elution for 2 min, the gradient of mobile phase A changed linearly by 5% within 6 min, this gradient was maintained for 7 min isocratic elution, and finally the gradient of mobile phase A was increased to 95% within 0.1 min.

[0040] Selective reaction monitoring (SRM) was used for detection. In positron emission tomography (PET) mode, the precursor and daughter ions detected were oxidized coenzyme I (m / z 664.1→136.0) and reduced coenzyme I (m / z 666.2→302.3), respectively. In anion mode, the precursor and daughter ions of dihydroxyacetone phosphate were detected (m / z 168.8→97.3). LC-MS / MS data were analyzed using Xcalibur 2.0 software. The results are shown in Figure 1. It is evident that the organometallic macrocyclic compound hemin significantly increased the intracellular oxidized coenzyme I / reduced coenzyme I ratio, with the best effect observed at a hemin concentration of 3 μg / mL.

[0041] Example 2

[0042] This example investigated the effects of organometallic macrocyclic compounds on mitochondrial function. Antimycin A was added to the cell culture medium to a final concentration of 1 μM to construct a cell model of mitochondrial damage. The ability of intracellular mitochondria to synthesize ATP before and after administration of the organometallic macrocyclic compound hemin chloride was measured (expressed by the ratio of oxidized coenzyme I to reduced coenzyme I). The results are shown in Figure 2, demonstrating that the organometallic macrocyclic compound hemin chloride can restore mitochondrial function.

[0043] Example 3

[0044] This study investigated the effects of organometallic macrocyclic compounds on increasing the level of oxidized coenzyme I in mouse liver. LC-MS / MS was used to detect changes in oxidized coenzyme I levels in mouse liver before and 24 hours after a single oral administration of different concentrations of organometallic macrocyclic compounds (ferroheme, heme chloride, and methylcobalamin). The results, normalized to nicotinamide ribose, are shown in Figure 3. The results confirm that organometallic macrocyclic compounds can effectively increase the ratio of oxidized to reduced coenzyme I in mouse tissues, with ferroheme showing the best effect.

[0045] Example 4

[0046] This study investigated the effect of organometallic macrocyclic compounds on the lifespan of mice. The organometallic macrocyclic compound Ferroheme was dissolved in drinking water and administered orally to 17-month-old male and female mice. The time of death of the mice was continuously recorded. The results are shown in Figure 4. The results showed that the lifespan of older mice administered organometallic macrocyclic compounds was significantly increased.

[0047] Example 5

[0048] This study investigated the effects of organometallic macrocyclic compounds on improving the health of mice. Using a rotarod experiment, we examined the muscle endurance and balance of aging mice after long-term intake of the organometallic macrocyclic compound Ferroheme (approximately 10 mg / kg) through drinking water for 300 days. The results are shown in Figure 5, confirming that organometallic macrocyclic compounds can effectively improve the health of aging mice.

[0049] The present invention has been described in detail above with reference to specific embodiments and exemplary examples; however, these descriptions should not be construed as limiting the present invention. Those skilled in the art will understand that various equivalent substitutions, modifications, or improvements can be made to the technical solutions and embodiments of the present invention without departing from the spirit and scope of the invention, and all such modifications and improvements fall within the scope of the present invention. The scope of protection of the present invention is defined by the appended claims.

Claims

1. The application of an organometallic macrocyclic compound molecule in catalytically increasing intracellular oxidized coenzyme I levels and in anti-aging products.

2. The application as described in claim 1, characterized in that, The structural formula of the organometallic macrocyclic compound molecule is: Where M is a metal, R1-R 12 Independently selected from chain alkyl groups, cyclic alkyl groups, hydrogen, aromatic groups, or functional groups containing oxygen, nitrogen, or sulfur, R 13 It is selected from hydrogen, or functional groups containing oxygen, nitrogen, sulfur, or halogen.

3. The application as described in claim 2, characterized in that, M is Cu, Fe, Co, Ni, Mn, Zn, Pt, Pb, V, Ag, Sn, Gd, Ga, Cd, Ir, Rh, or In.

4. The application as described in claim 1, characterized in that, The catalytic enhancement of intracellular oxidized coenzyme I levels specifically involves: catalyzing the conversion of intracellular reduced coenzyme I into oxidized coenzyme I, thereby increasing intracellular oxidized coenzyme I levels.

5. The application as described in claim 1, characterized in that, The cells include cardiomyocytes, hepatocytes, lung cells, fibroblasts, epithelial cells, and immune cells.

6. The application as described in claim 5, characterized in that, The immune cells include macrophages, T cells, NK cells, and B cells.

7. The application as described in claim 1, characterized in that, The organometallic macrocyclic compound molecules improve mitochondrial metabolic function by increasing intracellular levels of oxidized coenzyme I.

8. The application as described in claim 7, characterized in that, The functions of mitochondria include ATP synthesis and aerobic respiration.

9. The application as described in claim 1, characterized in that, The products include oxidized coenzyme I supplements.

10. The application as described in claim 1, characterized in that, The products include anti-aging drugs or health supplements.

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