Application of carboline compound in preparation of product for reducing accumulation of advanced glycosylation end product

Abstract

The invention discloses application of carboline compounds in preparation of products for reducing accumulation of advanced glycosylation end products, and belongs to the technical field of preparation of anti-saccharification products. The carboline compound not only can reduce the generation of AGEs in a BSA-MGO pure in-vitro AGEs generation model, but also can effectively reduce the level of AGEs in a glucose-induced intracellular AGEs generation model, has a remarkable AGEs inhibition effect, has no toxic effect on cells, has good safety, and can meet the application requirements of anti-saccharification products.

Description

Technical Field

[0001] The present application relates to the technical field of preparation of anti-glycation products, and in particular to the application of carboline compounds in the preparation of products for reducing the accumulation of advanced glycation end products. Background Art

[0002] Advanced glycation end products (AGEs) are a class of compounds formed by non-enzymatic reactions between proteins, lipids, and carbohydrates. Their accumulation is closely associated with various chronic diseases and the aging process. The accumulation of AGEs in the body disrupts cellular structure and function, leading to a hardening of the extracellular matrix, particularly in the skin, blood vessels, eyes, and nervous system. Their cross-linking effects on collagen can lead to decreased skin elasticity, wrinkle formation, vascular hardening, and other degenerative changes. AGEs can induce a range of pathological conditions, including free radical formation, inflammatory responses, antioxidant resistance, abnormal mitochondrial metabolism, and vasoconstriction. They are highly correlated with the development and progression of numerous metabolic diseases, including diabetes, metabolic syndrome, obesity, hypertension, arteriosclerosis, Alzheimer's disease, and peripheral neuropathy.

[0003] Current research has identified a variety of natural and synthetic compounds with the potential to reduce the formation of AGEs or accelerate their clearance. For example, antioxidants such as vitamins C and E can neutralize free radicals and inhibit the formation of AGEs. Natural plant extracts such as resveratrol, quercetin, and catechins reduce the accumulation of AGEs by inhibiting glycation and blocking the binding of sugars to proteins.

[0004] In order to fight glycation, common anti-sugar strategies mainly include the following aspects. First, control the sugar intake in the diet and reduce the intake of high-sugar, high-calorie foods, especially refined sugars. Secondly, the intake of foods rich in antioxidants, such as green tea, polyphenols, vitamin C and vitamin E, can reduce the production of glycation products. Increasing foods rich in dietary fiber, such as whole grains, vegetables and fruits, can also help maintain stable blood sugar and prevent glycation reactions. Finally, maintaining good living habits, such as regular exercise, adequate sleep and avoiding excessive stress, can help reduce the risk of glycation reactions in the body.

[0005] However, existing anti-glycation substances, such as plant extracts and antioxidants, although able to slow down glycation, have limited effects in reducing the accumulation of AGEs, especially in terms of stability and human safety during long-term use. Summary of the Invention

[0006] Based on this, the main purpose of this application is to provide the use of carboline compounds in the preparation of products that reduce the accumulation of advanced glycation end products. The carboline compounds have significant AGEs inhibitory effects, are non-toxic to cells, have good safety, and can meet the application requirements of anti-glycation products.

[0007] In a first aspect of the present application, a carboline compound is provided for use in preparing a product for reducing accumulation of advanced glycation end products. The carboline compound is a compound represented by Formula I:

[0008] (Formula I);

[0009] Wherein, R1 is a C2-C5 alkyl group substituted by a hydroxyl group; and R2 is -COOH.

[0010] In some embodiments, the C2-C5 alkyl group is a straight chain alkyl group.

[0011] In some embodiments, the C2-C5 alkyl group substituted with a hydroxyl group is -CH(OH)-R3; R3 is a C1-C4 straight-chain alkyl group or a C1-C4 straight-chain alkyl group substituted with a hydroxyl group at its terminal position.

[0012] In some embodiments, R3 is -CH3 or -CH2-OH.

[0013] In some embodiments, the carboline compound is selected from at least one of the following compounds:

[0014] 、 .

[0015] In some embodiments, the product for reducing the accumulation of advanced glycation end products is a product for preventing and / or treating diseases associated with the accumulation of AGEs.

[0016] In some embodiments, the diseases associated with the accumulation of AGEs include aging and / or degenerative diseases.

[0017] In some embodiments, the degenerative disease comprises at least one of diabetes, cardiovascular disease, metabolic syndrome, obesity, hypertension, arteriosclerosis, neurodegenerative disease, orthopedic disease and female ovarian dysfunction.

[0018] In some embodiments, the neurodegenerative disease comprises Alzheimer's disease and / or peripheral neuropathy.

[0019] In some embodiments, the product that reduces accumulation of advanced glycation end products comprises a cosmetic, a food, or a pharmaceutical.

[0020] In some embodiments, the product for reducing accumulation of advanced glycation end products is administered orally and / or topically.

[0021] In some embodiments, the dosage form of the product includes a solid dosage form, a semisolid dosage form, or a liquid dosage form.

[0022] In a second aspect, the present application provides a product for reducing the accumulation of advanced glycation end products, including a carboline compound, wherein the carboline compound has a structure represented by Formula I:

[0023] (Formula I);

[0024] wherein R1 is a C2-C5 alkyl group substituted with a hydroxyl group; R2 is -COOH;

[0025] Optionally, the product for reducing accumulation of advanced glycation end products further comprises excipients.

[0026] In a third aspect, the present application provides a product for preventing and / or treating diseases associated with the accumulation of AGEs, comprising a carboline compound having a structure represented by Formula I:

[0027] (Formula I);

[0028] wherein R1 is a C2-C5 alkyl group substituted with a hydroxyl group; R2 is -COOH;

[0029] Optionally, the product for preventing and / or treating diseases associated with the accumulation of AGEs further comprises excipients.

[0030] Compared with the prior art, this application has the following beneficial effects:

[0031] 1. The carboline compounds (R)-(+)-Dichotomine A and Dichotomine B can not only reduce AGEs formation in the BSA-MGO pure in vitro AGEs formation model, but also effectively reduce AGEs levels in the glucose-induced intracellular AGEs formation model. They have significant AGEs inhibitory effects and can be used to prevent, treat or improve diseases related to the accumulation of AGEs.

[0032] 2. Carboline compounds have no toxic effects on cells and have good safety. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] In order to more clearly illustrate the technical solutions in the embodiments of the present application or the prior art, the following briefly introduces the drawings required for the embodiments or the prior art description. The drawings are only used to illustrate the preferred embodiments and are not considered to limit the present application. The same reference symbols are used throughout the drawings to represent the same components. In the drawings:

[0034] Figure 1 The inhibition rate of (R)-(+)-Dichotomine A on AGEs;

[0035] Figure 2 This is the inhibition rate of Dichotomine B on AGEs. DETAILED DESCRIPTION

[0036] To make the objectives, technical solutions, and advantages of this application more clear and to provide a more thorough and comprehensive understanding of the disclosure of this application, the following will provide a clear and complete description of the technical solutions of this application in conjunction with the specific embodiments of this application and the corresponding drawings. The described embodiments are only part of the embodiments of this application, not all of them.

[0037] The following is a detailed description of the implementation of this application in conjunction with the accompanying drawings. This embodiment is implemented based on the technical solution of this application, and provides a detailed implementation method and specific operation process, but the scope of protection of this application is not limited to the following embodiment.

[0038] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art to which this application pertains. The terms used herein in the specification of this application are for the purpose of describing specific embodiments only and are not intended to limit this application.

[0039] the term

[0040] Unless otherwise specified or incompatible herewith, the terms and phrases used herein shall have the following meanings:

[0041] In this application, "a plurality of", "a plurality of", "at least one", etc., unless otherwise specified, refer to a number greater than or equal to 2. For example, "one or more" or "at least one" means one or more than or equal to two.

[0042] In this application, the terms "preferable", "particularly", etc. are used for descriptive purposes to indicate differences in content, but should not be understood as limiting the scope of protection of this application.

[0043] The term "alkyl" refers to a saturated hydrocarbon containing primary (normal) carbon atoms, secondary carbon atoms, tertiary carbon atoms, quaternary carbon atoms, or combinations thereof. Phrases containing this term, such as "C1-C5 alkyl," refer to an alkyl group containing 1 to 5 carbon atoms, each occurrence of which may independently be C1 alkyl, C2 alkyl, C3 alkyl, C4 alkyl, or C5 alkyl. Suitable examples include, but are not limited to, methyl (Me, -CH3), ethyl (Et, -CH2CH3), 1-propyl (n-Pr, n-propyl, -CH2CH2CH3), 2-propyl (i-Pr, i-propyl, -CH(CH3)2), 1-butyl (n-Bu, n-butyl, -CH2CH2CH2CH3), 2-methyl-1-propyl (i-Bu, i-butyl, -CH2CH(CH3)2), 2-butyl (s-Bu, s-butyl, -CH(CH3)CH2CH3), 2-methyl-2-propyl (t- Bu, t-butyl, -C(CH3)3), 1-pentyl (n-pentyl, -CH2CH2CH2CH2CH3), 2-pentyl (-CH(CH3)CH2CH2CH3), 3-pentyl (-CH(CH2CH3)2), 2-methyl-2-butyl (-C(CH3)2CH2CH3), 3-methyl-2-butyl (-CH(CH3)CH(CH3)2), 3-methyl-1-butyl (-CH2CH2CH(CH3)2) and 2-methyl-1-butyl (-CH2CH(CH3)CH2CH3).

[0044] In this application, the technical features described in an open manner include closed technical solutions composed of the listed features, and also include open technical solutions containing the listed features.

[0045] In a first aspect of the present application, a carboline compound is provided for use in preparing a product for reducing accumulation of advanced glycation end products. The carboline compound is a compound represented by Formula I:

[0046] (Formula I);

[0047] Wherein, R1 is a C2-C5 alkyl group substituted by a hydroxyl group; and R2 is -COOH.

[0048] In some embodiments, the C2-C5 alkyl group is a straight chain alkyl group.

[0049] In some embodiments, the C2-C5 alkyl group substituted with a hydroxyl group is -CH(OH)-R3; R3 is a C1-C4 straight-chain alkyl group or a C1-C4 straight-chain alkyl group substituted with a hydroxyl group at its terminal position.

[0050] It is understood that the hydroxy-substituted C2-C5 alkyl group may be substituted by one or more hydroxy groups, preferably by one or two hydroxy groups, wherein one hydroxy group is substituted on the first carbon atom of R1, and when substituted by multiple hydroxy groups, at least one hydroxy group is a terminal substitution.

[0051] In some embodiments, R3 is -CH3 or -CH2-OH.

[0052] In some embodiments, the carboline compound is selected from at least one of the following compounds:

[0053] 、 .

[0054] Carboline compound (R)-(+)-Dichotomine A, molecular formula C 14 H 12 N2O3, carboline compound Dichotomine B, molecular formula C 14 H 12 N2O4, the structural formulas of the two are as follows:

[0055] 、 .

[0056] (R)-(+)-Dichotomine A and Dichotomine B are β-carboline alkaloids. Studies have shown that they can attenuate LPS / ATP-induced neuroinflammatory responses in BV2 microglia, possibly through the TLR4 / MyD88-mTOR signaling pathway and autophagy. In this application, (R)-(+)-Dichotomine A and Dichotomine B not only reduce AGE formation in a pure BSA-MGO in vitro AGE formation model, but also effectively reduce AGE levels in a glucose-induced intracellular AGE formation model, demonstrating significant AGE inhibition and no cellular toxicity. These compounds have a good safety profile and meet the application requirements of anti-glycation products.

[0057] In some embodiments, the product for reducing the accumulation of advanced glycation end products is a product for preventing and / or treating diseases associated with the accumulation of AGEs.

[0058] In some embodiments, the diseases associated with the accumulation of AGEs include aging and / or degenerative diseases.

[0059] In some embodiments, the degenerative disease comprises at least one of diabetes, cardiovascular disease, metabolic syndrome, obesity, hypertension, arteriosclerosis, neurodegenerative disease, orthopedic disease and female ovarian dysfunction.

[0060] In some embodiments, the neurodegenerative disease comprises Alzheimer's disease and / or peripheral neuropathy.

[0061] It can be understood that the carboline compounds can not only reduce AGEs production in the BSA-MGO pure in vitro AGEs production model, but can also effectively reduce AGEs levels in the glucose-induced intracellular AGEs production model, and have a significant inhibitory effect on AGEs production, which can prevent the accumulation of AGEs. Therefore, they are highly feasible for preventing, treating or ameliorating diseases caused by AGEs accumulation, such as aging, as well as degenerative diseases such as diabetes, cardiovascular disease, metabolic syndrome, obesity, hypertension, arteriosclerosis, neurodegenerative diseases, orthopedic diseases and female ovarian dysfunction.

[0062] In some embodiments, the product that reduces accumulation of advanced glycation end products comprises a cosmetic, a food, or a pharmaceutical.

[0063] In some embodiments, the product for reducing accumulation of advanced glycation end products is administered orally and / or topically.

[0064] It is understandable that the carboline compounds are not only suitable for oral administration and enter the human body through gastrointestinal absorption to treat diseases or regulate the body's physiological functions, but are also suitable for external use, acting locally on the affected area to improve symptoms, and can therefore be used in cosmetics, food or medicine.

[0065] In some embodiments, the dosage form of the product includes a solid dosage form, a semisolid dosage form, or a liquid dosage form.

[0066] Specifically, the solid dosage forms include tablets, capsules, pills, and granules; the semisolid dosage forms include ointments, gel ointments, and patches; and the liquid dosage forms include solutions, syrups, emulsions, and tinctures. When the product for reducing the accumulation of advanced glycation end products is administered in a liquid dosage form, the effective concentration can be 0.1-100 μM, preferably 0.4-100 μM, and specifically 0.4 μM, 1 μM, 3 μM, 5 μM, 10 μM, 20 μM, 30 μM, 40 μM, 50 μM, 60 μM, 70 μM, 80 μM, 90 μM, 100 μM, and the like.

[0067] In a second aspect, the present application provides a product for reducing the accumulation of advanced glycation end products, including a carboline compound, wherein the carboline compound has a structure represented by Formula I:

[0068] (Formula I);

[0069] wherein R1 is a C2-C5 alkyl group substituted with a hydroxyl group; R2 is -COOH;

[0070] Optionally, the product for reducing accumulation of advanced glycation end products further comprises excipients.

[0071] In a third aspect, the present application provides a product for preventing and / or treating diseases associated with the accumulation of AGEs, comprising a carboline compound having a structure represented by Formula I:

[0072] (Formula I);

[0073] wherein R1 is a C2-C5 alkyl group substituted with a hydroxyl group; R2 is -COOH;

[0074] Optionally, the product for preventing and / or treating diseases associated with the accumulation of AGEs further comprises excipients.

[0075] It is understandable that the carboline compound can be prepared into different dosage forms by adding excipients, and the excipients may specifically include carriers, fillers, diluents, excipients, solvents, functional additives, coating materials or encapsulation materials, etc.

[0076] Unless otherwise specified, the raw materials used in the following experiments can be purchased from the market.

[0077] The following are specific examples.

[0078] 1. The effects of different anti-glycation and anti-aging substances on the amount of AGEs produced in the BSA-MGO model.

[0079] Conventional AGEs have fluorescent properties. This experimental example uses the principle that methyl glycol and bovine serum albumin undergo glycation reaction in vitro to form AGEs for modeling. After adding anti-glycation and anti-aging substances, the AGE inhibition rate is calculated by fluorescence quantitative method to evaluate its anti-glycation efficacy.

[0080] 1. Test materials and instruments

[0081] 1.1 Test Reagents

[0082] Bovine serum albumin (BSA), methylglyoxal (MGO), and glucose: analytical grade, Sigma, USA.

[0083] 1.2 Test samples

[0084] Anti-glycation and anti-aging substances: (R)-(+)-Dichotomine A, CAS#: 1449383-70-3; Dichotomine B, CAS#: 755036-41-0; Aminoguanidine, CAS#: 1937-19-5.

[0085] 1.3 Test instruments

[0086] Microplate reader (Synergy H1), Bio-Tek, USA.

[0087] 2. Test methods

[0088] 2.1 Establishment of in vitro protein AGE formation system

[0089] Bovine serum albumin (BSA) and methylglyoxal (MGO) were prepared at 10 mg / ml and 25 mM concentrations, respectively, in phosphate buffer (10 mM, pH 7.4). This formed a BSA-MGO reaction system, which served as a model for AGE formation. The OD values ​​of the formed AGEs were measured using a microplate reader (Synergy H1, Bio-Tek) at a detection wavelength of 370 nm and a reference wavelength of 440 nm.

[0090] 2.2 Determination of inhibition of AGE formation in vitro

[0091] Mixed incubation solutions prepared in Examples 1A-3A, Examples 1B-3B, and Comparative Examples 1-2 were prepared using a solution containing 10 mg / ml BSA and 25 mM MGO. A control (blank) solution consisted of a BSA-MGO reaction system without the test substance. All solutions were sterile-filtered using a 0.2 μm filter and aliquoted into 1.5 mL conical centrifuge tubes.

[0092] Table 1: Contents of anti-glycation and anti-aging substances in different examples and comparative examples

[0093]

[0094] Then, 100 μl of the reaction solution was placed in a black 96-well microplate, and the fluorescence value was measured on day 0. The OD value of the formed AGEs was measured using a microplate reader (Synergy H1, Bio-Tek) (detection wavelength 370 nm, reference wavelength 440 nm).

[0095] The remaining reaction solution was placed in a 37°C incubator for 4 days. On the fourth day, 100 μl of the reaction solution was transferred to a black 96-well microtiter plate and the fluorescence value was measured on the fourth day. The OD value of the formed AGEs was measured using a microtiter plate reader (Synergy H1, Bio-Tek) (detection wavelength: 370 nm, reference wavelength: 440 nm).

[0096] 2.3 Data processing and statistical analysis methods (each experiment was repeated at least three times)

[0097] The AGE inhibition rate was calculated using the formula;

[0098] AGE inhibition rate = (CT) / C × 100%;

[0099] Wherein, C is the fluorescence intensity of the control group (blank case), that is, the fluorescence intensity of the BSA-MGO reaction system in the absence of anti-glycation and anti-aging substances; T is the fluorescence intensity of the BSA-MGO reaction system in the presence of anti-glycation and anti-aging substances.

[0100] 3. Test results

[0101] The inhibitory effect of the test samples on AGEs, such as Figure 1-2 and as shown in Table 2.

[0102] Table 2: AGE inhibition rates in Examples, Comparative Examples, and Blank Examples

[0103]

[0104] Through Figure 1-2 The AGEs inhibition rate was compared with the results in Table 2. It was found that the AGEs inhibition rate of aminoguanidine at 1 mM was 9.24%, and the AGEs inhibition rate at 10 mM was 51.84%.

[0105] As the concentration of (R)-(+)-Dichotomine A increased at 1μM, 3μM, and 10μM, the AGEs inhibition rate increased concentration-dependently, reaching 28.05%, 48.94%, and 68.85%, respectively. The AGEs inhibition effect was higher than that of 1mM aminoguanidine.

[0106] As the concentration of Dichotomine B increased at 1μM, 3μM, and 10μM, the AGEs inhibition rate gradually increased in a concentration-dependent manner, reaching 31.41%, 37.65%, and 59.30%, respectively. The AGEs inhibition effect was higher than that of 1mM aminoguanidine.

[0107] Furthermore, when the drug concentration of (R)-(+)-Dichotomine A or Dichotomine B was higher than 10 μM, the AGEs inhibition effect was higher than that of 10 mM aminoguanidine.

[0108] The above results prove that in the in vitro BSA-MGO system, (R)-(+)-Dichotomine A and Dichotomine B can significantly inhibit the formation of AGEs, and this inhibitory effect is concentration-dependent.

[0109] 2. Effects of different glucose concentrations on AGEs levels in HaCaT cells.

[0110] 1. Test materials and instruments

[0111] 1.1 Test Reagents

[0112] Human keratinocytes (HaCaT) and glucose: analytical grade, Sigma, USA. BCA (bicinchoninic acid) protein assay kit: Beyotime Biotechnology Co., Ltd. Human advanced glycation end products (AGEs) enzyme-linked immunosorbent assay kit: Wuhan Huamei Biotechnology Co., Ltd.

[0113] 1.2 Test instruments

[0114] Microplate reader (Synergy H1), Bio-Tek, USA.

[0115] 2. Test methods

[0116] 2.1 Construction of cell glycation model

[0117] This application uses glucose (D-Glucose) (MCE, HY-B0389, purity: ≥98.0%) as an inducer for the construction of a human keratinocyte (HaCaT) cell glycation model. Cells were grown to the logarithmic growth phase and plated, with the confluence of cells in a 6 cm dish reaching 60% on the second day.

[0118] 2.2 Effects of different glucose concentrations on HaCaT cell viability

[0119] HaCaT was seeded in a 96-well plate, and a concentration of 1.5×10 5 100 μl of a suspension of 100 μg / ml was added and cultured until the next day. The drug was diluted with culture medium to obtain Examples 4-6 and a blank as shown in Table 3, respectively, so that the final glucose concentrations were 100 mM, 150 mM, and 200 mM, respectively. After medium replacement, the cells were cultured for another 24 hours. 10 μl of Cell Counting Kit-8 (CCK-8, Dojindo) was added to each well of the 96-well plate and incubated for 1-2 hours. The OD value was measured at 450 nm using an enzyme microplate reader. CCK-8 is a kit based on WST-8 and widely used for rapid and highly sensitive detection of cell proliferation and cytotoxicity. In the presence of an electron coupling reagent, WST-8 can be reduced by certain dehydrogenases within the mitochondria to produce an orange-yellow formazan. The more and faster the cell proliferation, the darker the color; the greater the cytotoxicity, the lighter the color. For the same cells, the depth of the color is linearly related to the number of cells.

[0120] Table 3: Glucose content in different examples

[0121]

[0122] Table 4: Cell viability in Examples 4-6 and blank examples

[0123]

[0124] 2.3 Effects of different glucose concentrations on AGE levels in HaCaT cells

[0125] The drugs were diluted in complete culture medium (10% FBS (Gibco) + DMEM (Gibco)) to obtain Examples 4-6 and a blank as shown in Table 3, respectively, with final glucose concentrations of 100 mM, 150 mM, and 200 mM, respectively. After incubating the cells for 48 hours, cell samples were collected for BCA assay and intracellular AGE content determination, and the AGE content per unit protein was calculated.

[0126] The OD values ​​of the formed AGEs were measured using a microplate reader (Synergy H1, Bio-Tek) at a detection wavelength of 370 nm and a reference wavelength of 440 nm. BCA (bicinchoninic acid) was measured using techniques known in the art. Under alkaline conditions, divalent copper ions are reduced to monovalent copper ions by proteins. The monovalent copper ions interact with the unique BCA Solution A to produce a sensitive color reaction. This water-soluble complex exhibits strong absorbance at 562 nm. The absorbance and protein concentration show a good linear relationship over a wide range, allowing protein concentration to be estimated based on the absorbance. Results are shown in Table 5.

[0127] Table 5: Effects of different examples on AGEs levels in HaCaT cells

[0128]

[0129] According to the cell viability test results in Section 2.2 and microscopic observations, when the glucose concentration reached 200 mM, cells showed obvious cell death. Combined with Table 5, it can be seen that when the final glucose concentration reached 150 mM, the AGEs level increased to 1.45 times the normal level. Therefore, 150 mM glucose can be used as the induction condition for the cell glycation model.

[0130] 3. (R)-(+)-Dichotomine A and Dichotomine B can reduce the production of intracellular AGEs in a glucose-induced cell glycation model.

[0131] 1. Test materials and instruments

[0132] 1.1 Test Reagents

[0133] Human keratinocytes (HaCaT) and glucose: analytical grade, Sigma, USA. BCA (bicinchoninic acid) protein assay kit: Beyotime Biotechnology Co., Ltd. Human advanced glycation end products (AGEs) enzyme-linked immunosorbent assay kit: Wuhan Huamei Biotechnology Co., Ltd.

[0134] 1.2 Test instruments

[0135] Microplate reader (Synergy H1), Bio-Tek, USA.

[0136] 2. Test methods

[0137] 2.1 Construction of cell glycation model

[0138] This application uses glucose (D-Glucose) (MCE, HY-B0389, purity: ≥98.0%) as an inducer for the construction of a human keratinocyte (HaCaT) cell glycation model. Cells were grown to the logarithmic growth phase and plated, with the confluence of cells in a 6 cm dish reaching 60% on the second day.

[0139] 2.2 Effects of (R)-(+)-Dichotomine A and Dichotomine B on HaCaT cell viability

[0140] HaCaT was seeded in a 96-well plate, and a concentration of 1.5×10 5 / ml suspension 100μl, culture until the next day, add the complete culture medium containing 0.4μM, 2μM, 10μM (R)-(+)-Dichotomine A or Dichotomine B and continue to culture for 24h. Add 10μl Cell Counting Kit-8 (CCK-8, Dojindo) to each well of the 96-well plate and culture for 1-2h. The OD value is detected at 450nm using an enzyme microplate reader.

[0141] Table 6: Cell viability of different concentrations of (R)-(+)-Dichotomine A, Dichotomine B and blank samples

[0142]

[0143] 2.3 Effects of different glucose concentrations on AGE levels in HaCaT cells

[0144] The drugs were diluted in complete culture medium (10% FBS (Gibco) + DMEM (Gibco)) to obtain Examples 7A-9A, Examples 7B-9B, and a blank as shown in Table 7, respectively. The final concentrations of glucose were 150 mM, and the final concentrations of (R)-(+)-Dichotomine A and Dichotomine B were 0.4 μM, 2 μM, and 10 μM, respectively. After incubating the cells for 48 hours, cell samples were collected for BCA assay and intracellular AGE content, and the AGE content per unit protein was calculated.

[0145] The OD value of the formed AGEs was measured by a microplate reader (Synergy H1, Bio-Tek) (detection wavelength 370 nm, reference wavelength 440 nm).

[0146] Table 7: Contents of glucose and (R)-(+)-Dichotomine A and Dichotomine B in different examples

[0147]

[0148] Table 8: Effects of different examples on AGEs levels in HaCaT cells

[0149]

[0150] As can be seen from the table above, after treatment with 150 mM glucose, the intracellular AGEs level in the comparative example increased to 1.34 times that of the blank example (control group).

[0151] When glucose and (R)-(+)-Dichotomine A were treated simultaneously, the level of intracellular AGEs was significantly reduced compared with the group treated with glucose alone. When the concentration of (R)-(+)-Dichotomine A was 10 μM, the intracellular AGEs were only 0.70 times that of the blank case, indicating that (R)-(+)-Dichotomine A has a highly effective AGE inhibitory effect.

[0152] When glucose and Dichotomine B were treated simultaneously, the level of intracellular AGEs was significantly reduced compared to the group treated with glucose only. When the Dichotomine B concentration was 0.2 μM, the intracellular AGEs were only 0.66 times that of the blank case, indicating that Dichotomine B has a highly efficient AGEs inhibitory effect.

[0153] When (R)-(+)-Dichotomine A or Dichotomine B is used in the preparation of anti-glycation foods and anti-glycation health products, especially in beauty products, the effective concentrations of (R)-(+)-Dichotomine A and Dichotomine B are recommended to refer to the data recorded in the above examples.

[0154] The technical features of the above-mentioned embodiments can be combined arbitrarily. In order to make the description concise, not all possible combinations of the technical features in the above-mentioned embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0155] The above-described embodiments merely represent several implementation methods of the present application. While the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the present invention. It should be noted that a person skilled in the art could make various modifications and improvements without departing from the spirit of the present application, all of which fall within the scope of protection of the present application. Therefore, the scope of protection of the present patent application shall be determined by the appended claims.

Claims

1. Use of a carboline compound in the preparation of a product for reducing accumulation of advanced glycation end products, characterized in that: The carboline compound is a compound represented by formula I: (Formula I); Wherein, R1 is a C2-C5 alkyl group substituted by a hydroxyl group; and R2 is -COOH.

2. The use according to claim 1, characterized in that The C2-C5 alkyl group is a straight-chain alkyl group.

3. The use according to claim 1, characterized in that The C2-C5 alkyl group substituted by a hydroxyl group is -CH(OH)-R3; R3 is a C1-C4 straight-chain alkyl group or a C1-C4 straight-chain alkyl group substituted by a hydroxyl group at the terminal.

4. The use according to claim 3, characterized in that R3 is -CH3 or -CH2-OH.

5. The use according to claim 4, characterized in that The carboline compound is selected from at least one of the following compounds: 、 。 6. The use according to any one of claims 1 to 5, characterized in that: The product for reducing the accumulation of advanced glycation end products is a product for preventing and / or treating diseases associated with the accumulation of AGEs.

7. The use according to claim 6, characterized in that The diseases associated with the accumulation of AGEs include aging and / or degenerative diseases; Optionally, the degenerative disease includes at least one of diabetes, cardiovascular disease, metabolic syndrome, obesity, hypertension, arteriosclerosis, neurodegenerative disease, orthopedic disease and female ovarian dysfunction; Optionally, the neurodegenerative disease comprises Alzheimer's disease and / or peripheral neuropathy.

8. The use according to any one of claims 1 to 5, characterized in that: The product for reducing the accumulation of advanced glycation end products includes cosmetics, food or medicine; and / or the administration of the product for reducing accumulation of advanced glycation end products includes oral administration and / or topical administration; And / or the dosage form of the product includes a solid dosage form, a semisolid dosage form or a liquid dosage form.

9. A product for reducing accumulation of advanced glycation end products, characterized in that Including carboline compounds, the carboline compound structure is a compound shown in formula I: (Formula I); Wherein, R1 is a C2-C5 alkyl group substituted with a hydroxyl group; R2 is -COOH; Optionally, the product for reducing accumulation of advanced glycation end products further comprises auxiliary materials.

10. A product for preventing and / or treating diseases associated with the accumulation of AGEs, characterized in that Including carboline compounds, the carboline compound structure is a compound shown in formula I: (Formula I); Wherein, R1 is a C2-C5 alkyl group substituted with a hydroxyl group; R2 is -COOH; Optionally, the product for preventing and / or treating diseases associated with the accumulation of AGEs further comprises excipients.

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