A ketone compound and its anti-glycation application

Abstract

The present invention discloses use of a compound of the following formula I or a salt or deuterated product thereof in the preparation of an anti-glycation agent, wherein R1 is a hydroxyl group or hydrogen, R2 is a methoxy group or H, R3 is a hydroxyl group or hydrogen, R4 is a methoxy group, a hydroxyl group or H, R5 is an isopentenyl group, a hydroxyl group or hydrogen, R6 is a methoxy group or a hydroxyl group, and R7 is an isopentenyl group, a methoxy group, a hydroxyl group or hydrogen; wherein R5 and R7 are not both isopentenyl groups. This type of anti-glycation active molecule is a natural #imgabs0# ketone compound (with only one isopentenyl side chain), which is derived from synthetic or natural plants and has good safety. In the experiment of inhibiting the formation of AGEs, compared with the Yangshen drug aminoguanidine (AG) and 1,7-dihydroxy-2-methoxy-3-(3-methylbut-2-enyl) #imgabs1# ketone (1,7-dihydroxy-2-methoxy-3-(3-methylbut-2-enyl)xanthone), the anti-glycation effect is significant; the anti-glycation inhibition rate of the natural #imgabs2# ketone compound protected by the present invention (final concentration of 250 μM) is between 18%-53%, which is much higher than the anti-glycation inhibition rate (17%) of Yangshen AG at the same final concentration (250 μM). #imgabs3#

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to a Ketone compounds and their anti-glycation applications. Background Art

[0002] Protein glycation occurs slowly and continuously throughout the life cycle of an organism, leading to the continuous accumulation of advanced glycation end products (AGEs) and their involvement in the pathogenesis of age-related diseases.

[0003] The process of protein glycation is complex. The process is initiated by the reaction of the following substances: the carbonyl group of reducing sugars (such as glucose, fructose or ribose, etc.) forms Schiff bases with proteins with free amino groups, and then undergoes Amadori rearrangement. The resulting Amadori products are further degraded to form active dicarbonyl compounds such as methylglyoxal, glyoxal and 3-deoxyglucose ketone. The dicarbonyl compounds then react with a single amino group on the side chain of lysine or arginine, or the sulfhydryl group of cysteine ​​in the protein, or cross-link two of the residues. These reactions lead to the formation of various cross-linked adducts, collectively referred to as advanced glycation end products (AGEs). Amadori products can also directly rearrange to form AGEs.

[0004] AGEs tend to bind to proteins with high lysine and arginine unit content and long half-life, such as collagen, serum albumin, elastin, myelin, low-density lipoprotein, plasminogen activator, fibrinogen, etc. These glycated proteins lose their function and are resistant to proteolytic clearance by proteases. The accumulation of AGEs in the extracellular matrix leads to its abnormal cross-linking and reduced vascular elasticity, leading to atherosclerosis. In addition, AGEs receptors (RAGE) are expressed on many cell types. AGE-RAGE interactions lead to the production of reactive oxygen species, inflammation, angiogenesis, and proliferation through various signal transduction pathways. The activation of RAGE stimulates the phosphorylation of extracellular signal-regulated kinases, ultimately leading to the stimulation of the transcription factor NF-κB and the production of proinflammatory cytokines. In addition, the stimulation of RAGE leads to the activation of the transforming growth factor (TGF-β) pathway and induces the overexpression of vascular endothelial growth factor. The formation of intracellular AGEs can also lead to nitric oxide quenching and impaired growth factor function.

[0005] Protein glycation that occurs in heat-treated foods is called the Maillard reaction, and the AGEs produced are introduced into the human body through the exogenous route of diet. AGEs are naturally present in uncooked animal-derived foods, and cooking forms new AGEs in these foods. Nε-(carboxymethyl)lysine (CML) levels have been found to be a useful marker of AGE content in the diet and have been selected to characterize the AGE load of foods. A survey of the CML content of 549 foods showed that grilling, baking, scorching and frying accelerate the formation of new AGEs. These modified proteins are not digested but can be fermented by colon bacteria. After fermentation, AGEs form toxic compounds that have negative effects on human health.

[0006] Skin aging is a complex process, which is manifested by the decline of body function and regeneration capacity. Skin aging includes exogenous and intrinsic aging. Exogenous aging is mainly related to ultraviolet radiation, while intrinsic aging is an inevitable physiological change that occurs over time. In recent years, studies have shown that intrinsic aging is closely related to skin glycation. Glycation is a non-enzymatic reaction between nucleic acids, proteins and lipids and sugars, which ultimately produces stable and irreversible products, namely advanced glycation end products (AGEs). Skin glycation acts on the body through the following pathways, leading to the occurrence of aging. First, the extracellular matrix (ECM) is one of the main targets of glycation. Collagen reacts with sugar to produce AGEs. The AGEs deposited in the matrix cross-link with the ECM, which damages the function of collagen and fibrin, resulting in a decrease in the elasticity and contractility of connective tissue. At the same time, glycated collagen inhibits the degradation of matrix metalloproteinases (MMPs), accelerates the degradation of collagen and reduces the synthesis of functional collagen. In addition, AGEs have the characteristics of yellow-dark color, which plays a key role in the process of skin color changing to dark yellow.

[0007] In short, the accumulation of AGEs in the body, especially on the skin, will cause cross-linking damage to proteins and binding to specific receptors, causing many skin problems, leading to dull skin, wrinkles, decreased collagen fiber elasticity, dark brown spots, and even promoting the production of matrix metalloproteinases that can decompose collagen and elastin. These factors will eventually lead to problems such as accelerated skin aging.

[0008] Although some existing oral medicines, health products, functional foods, and skin care products have anti-glycation effects, they cannot solve the changes in the aging level in the body, and lack dietary protein anti-glycation active ingredients. Therefore, there is an urgent need for safe and effective active ingredients for human anti-glycation and dietary protein anti-glycation as active additives for cosmetics, health products, functional foods, medicines and other products that maintain human health.

[0009] The prior art does not find the use of the compound or its salt or deuterated product required by the present invention in the preparation of an anti-glycation agent. Summary of the invention

[0010] Based on this, the present invention provides a use of a compound of the following formula I or a salt or deuterated product thereof in the preparation of an anti-glycation agent:

[0011]

[0012] R1 is hydroxyl or hydrogen, R2 is methoxy or H, R3 is hydroxyl or hydrogen, R4 is methoxy, hydroxyl or H, R5 is isopentenyl, hydroxyl or hydrogen, R6 is methoxy or hydroxyl, and R7 is isopentenyl, methoxy, hydroxyl or hydrogen;

[0013] wherein R5 and R7 are not isopentenyl at the same time;

[0014] The deuterated substance is a partially deuterated substance and / or a fully deuterated substance.

[0015] Furthermore, the number of isopentenyl groups in the compound of formula I or its salt or deuterated product is 1.

[0016] Furthermore, the number of hydroxyl groups in the compound of formula I or its salt or deuterated product is 2 to 3.

[0017] Furthermore, the number of methoxy groups in the compound of formula I or its salt or deuterated product is 1 to 2.

[0018] Further, the compound of formula I is any one of the following compounds:

[0019]

[0020] According to another aspect of the present invention, there is provided an anti-glycation composition, the anti-glycation composition comprising a compound of formula I or a salt or deuterated substance thereof;

[0021] The anti-glycation composition is a compound having anti-glycation activity that inhibits the generation of end products of non-enzymatic glycation reactions.

[0022] According to another aspect of the present invention, there is provided a use of the anti-glycation composition in the preparation of an anti-glycation agent, wherein the anti-glycation composition is a pharmaceutical composition, a composition for skin use, a health product and / or a functional food.

[0023] Furthermore, the pharmaceutical composition further comprises pharmaceutically acceptable excipients.

[0024] Furthermore, the auxiliary material is selected from one or more of the following: diluent, wetting agent, binder, disintegrant, inclusion agent, flavoring agent, sustained-release agent, retention agent, lubricant, dispersant, plasticizer, opacifier and antioxidant.

[0025] Furthermore, the dosage form of the pharmaceutical composition is a patch, emulsion, tablet, pill, capsule, powder, film, lozenge, granule or oral solution.

[0026] Furthermore, the skin composition further comprises one or more skin care substances for preventing and / or improving skin aging and / or accelerated skin aging, such as carnosine, acetylated hyaluronic acid, adenosine and / or kava extract.

[0027] Furthermore, the skin composition is in the form of an emulsion, an essence, a toner, a face cream or a face mask.

[0028] Furthermore, the skin aging and / or accelerated skin aging includes one or more of the following: dull skin, skin wrinkles, decreased elasticity of skin collagen fibers, and skin pigmentation patches.

[0029] Furthermore, the skin composition further comprises other auxiliary materials.

[0030] Furthermore, the auxiliary material is selected from one or more of the following: a thickener, a pH adjuster, a preservative, an antibacterial agent and a stabilizer.

[0031] Furthermore, the thickener is selected from one or more of the following: carbomer, sodium polyacrylate, xanthan gum and hydroxyethyl cellulose.

[0032] Further, the pH adjuster is selected from one or more of the following: triethanolamine, aminomethyl propanol, citric acid and lactic acid.

[0033] Further, the preservative is selected from one or more of the following: methylparaben, propylparaben, butylparaben, p-hydroxyacetophenone and phenoxyethanol.

[0034] Furthermore, the health care product and / or functional food is packaged food and food raw material.

[0035] Beneficial effects of the present invention:

[0036] These anti-glycation molecules are natural Ketone compounds (with only one isopentenyl side chain) are derived from synthetic or natural plants and have good safety. In the experiment of inhibiting AGEs production, they were compared with the Yangshen drug aminoguanidine (AG) and 1,7-dihydroxy-2-methoxy-3-(3-methylbut-2-enyl) Compared with the compound claimed to be protected by the present invention, the natural oxadiazole (1,7-dihydroxy-2-methoxy-3-(3-methylbut-2-enyl)xanthone) (with only one isopentenyl side chain, but the position of the isopentenyl side chain is different from that of the compound claimed to be protected by the present invention), the anti-glycation effect is significant; the natural oxadiazole (1,7-dihydroxy-2-methoxy-3-(3-methylbut-2-enyl)xanthone) The anti-glycation inhibition rate of ketone compounds (final concentration of 250 μM) was between 18% and 53%, which was much higher than the anti-glycation inhibition rate of Yangshen AG at the same final concentration (250 μM) (17%), and close to the anti-glycation inhibition rate of Yangshen AG at a higher final concentration (1 mM) (56%); at the same concentration (250 μM) 1,7-dihydroxy-2-methoxy-3-(3-methylbut-2-enyl) The anti-glycation inhibition rate of ketone is about 1.5%. BRIEF DESCRIPTION OF THE DRAWINGS

[0037] In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the drawings required for use in the description of the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without exceeding the scope of protection required by the present invention.

[0038] Figure 1 is the isopentenyl group of Example 1 Schematic diagram of the screening results of the anti-glycation activity (AGEs inhibition rate %) of ketone compounds. DETAILED DESCRIPTION

[0039] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative work are within the scope of protection of the present invention.

[0040] Unless otherwise indicated, all technical and scientific terms and abbreviations used herein have the meanings commonly understood by those of ordinary skill in the field of the invention or the field in which the terms are used. Although any methods, conditions, substances or materials similar to or equivalent to those disclosed herein may be used in the practice of the present invention, preferred methods, conditions, substances or materials are described herein.

[0041] The present invention is intended to encompass all alternatives, variations and equivalents that may be included in the present invention as defined by the claims. Those skilled in the art will recognize many methods and materials similar or equivalent to those described herein that can be used in the practice of the present invention. The present invention is in no way limited to the methods and materials described.

[0042] The following definitions shall apply to the present invention unless otherwise indicated. For purposes of the present invention, chemical elements are defined according to the Periodic Table of the Elements, CAS version and Handbook of Chemical Physics, 75th Ed., 1994. In addition, general principles of organic chemistry are found in "Organic Chemistry", Thomas Sorrell, University Science Books, Sausalito: 1999, and "March's Advanced Organic Chemistry", Michael B. Smith and Jerry March, John Wiley & Sons, New York: 2007, all of which are hereby incorporated by reference.

[0043] As used in the specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise.

[0044] In the present invention, the term "comprising" is synonymous with "including". As used herein, the terms "comprising", "including", "having", "containing" or any other variation thereof are intended to cover a non-exclusive inclusion. For example, a composition, process, method, article, or apparatus comprising the listed elements is not necessarily limited to only those elements, but may include other elements not expressly listed or inherent to such composition, process, method, article, or apparatus.

[0045] As described in the background technology section, the prior art does not find the use of the compound or its salt or deuterated product required by the present invention in the preparation of an anti-glycation agent. In order to solve the above problem, the present invention provides a compound of the following formula I or its salt or deuterated product for use in the preparation of an anti-glycation agent,

[0046]

[0047] R1 is hydroxyl or hydrogen, R2 is methoxy or H, R3 is hydroxyl or hydrogen, R4 is methoxy, hydroxyl or H, R5 is isopentenyl, hydroxyl or hydrogen, R6 is methoxy or hydroxyl, and R7 is isopentenyl, methoxy, hydroxyl or hydrogen;

[0048] wherein R5 and R7 are not isopentenyl at the same time;

[0049] The deuterated substance is a partially deuterated substance and / or a fully deuterated substance.

[0050] The compound of formula I or its salt or deuterated product of the present invention is a type of natural active molecule that can be used for anti-glycation, which can reduce the level of advanced glycation end products (AGEs), and is beneficial for preventing and treating skin problems caused by the accumulation of AGEs on the skin, including but not limited to dark yellow skin, wrinkles, decreased collagen fiber elasticity, dark brown spots, etc.

[0051] In a preferred embodiment, the number of isopentenyl groups in the compound of formula I or its salt or deuterated product is 1.

[0052] In a preferred embodiment, the number of hydroxyl groups in the compound of formula I or its salt or deuterated product is 2 to 3.

[0053] In a preferred embodiment, the number of methoxy groups in the compound of formula I or its salt or deuterated product is 1 to 2.

[0054] In a preferred embodiment, the compound of formula I is any one of the following compounds:

[0055]

[0056] According to another aspect of the present invention, there is provided an anti-glycation composition, the anti-glycation composition comprising a compound of formula I or a salt or deuterated substance thereof;

[0057] The anti-glycation composition is a compound having anti-glycation activity that inhibits the generation of end products of non-enzymatic glycation reactions.

[0058] According to another aspect of the present invention, there is provided a use of the anti-glycation composition in the preparation of an anti-glycation agent, wherein the anti-glycation composition is a pharmaceutical composition, a composition for skin use, a health product and / or a functional food.

[0059] In a preferred embodiment, the pharmaceutical composition further comprises a pharmaceutically acceptable excipient.

[0060] In a preferred embodiment, the auxiliary material is selected from one or more of the following: diluent, wetting agent, binder, disintegrant, inclusion agent, flavoring agent, sustained-release agent, retention agent, lubricant, dispersant, plasticizer, opacifier and antioxidant.

[0061] In a preferred embodiment, the pharmaceutical composition is in the form of a patch, emulsion, tablet, pill, capsule, powder, film, lozenge, granule or oral solution.

[0062] In a preferred embodiment, the skin composition further comprises one or more skin care substances for preventing and / or improving skin aging and / or accelerated skin aging, such as carnosine, acetylated hyaluronic acid, adenosine and / or kava extract.

[0063] In a preferred embodiment, the skin composition is in the form of an emulsion, an essence, a toner, a cream or a mask.

[0064] In a preferred embodiment, the skin aging and / or accelerated skin aging includes one or more of the following: dull skin, skin wrinkles, decreased elasticity of skin collagen fibers, and skin pigmentation patches.

[0065] In a preferred embodiment, the skin composition further comprises other auxiliary materials.

[0066] In a preferred embodiment, the auxiliary material is selected from one or more of the following: a thickener, a pH adjuster, a preservative, an antibacterial agent and a stabilizer.

[0067] In a preferred embodiment, the thickener is selected from one or more of the following: carbomer, sodium polyacrylate, xanthan gum and hydroxyethyl cellulose.

[0068] In a preferred embodiment, the pH adjuster is selected from one or more of the following: triethanolamine, aminomethyl propanol, citric acid and lactic acid.

[0069] In a preferred embodiment, the preservative is selected from one or more of the following: methylparaben, propylparaben, butylparaben, p-hydroxyacetophenone and phenoxyethanol.

[0070] In a preferred embodiment, the health care product and / or functional food is packaged food and food raw material.

[0071] According to another aspect of the present invention, there is provided a use of a compound of the following formula I or a salt or deuterated product thereof, or a composition comprising a compound of the following formula I or a salt or deuterated product thereof, in the preparation of an in vitro anti-glycation agent or an in vitro elastase inhibitor,

[0072]

[0073] R1 is hydroxyl or hydrogen, R2 is methoxy or H, R3 is hydroxyl or hydrogen, R4 is methoxy, hydroxyl or H, R5 is isopentenyl, hydroxyl or hydrogen, R6 is methoxy or hydroxyl, and R7 is isopentenyl, methoxy, hydroxyl or hydrogen;

[0074] wherein R5 and R7 are not isopentenyl at the same time;

[0075] The deuterated substance is a partially deuterated substance and / or a fully deuterated substance.

[0076] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental methods in the following examples without specifying specific conditions are usually carried out according to conventional conditions or conditions recommended by the manufacturer.

[0077] Unless otherwise defined, all professional and scientific terms used herein have the same meanings as those familiar to those skilled in the art. In addition, any methods and materials similar or equivalent to those described herein can be applied to the methods of the present invention. The preferred implementation methods and materials described herein are for demonstration purposes only.

[0078] The above features mentioned in the present invention or the features mentioned in the embodiments can be combined in any way. All the features disclosed in this patent specification can be used in combination with any combination form, and each feature disclosed in the specification can be replaced by any alternative feature that can provide the same, equal or similar purpose. Therefore, unless otherwise specified, the disclosed features are only general examples of equal or similar features.

[0079] Example 1

[0080] Anti-glycation test (inhibition of the formation of non-enzymatic glycation end products AGEs) experimental method

[0081] 1. Purpose of the experiment

[0082] The ability of the compounds to inhibit the synthesis of AGEs was evaluated by fluorescence detection of non-enzymatic glycation end products (AGEs).

[0083] 2. Experimental Principle

[0084] Advanced glycation end products (AGEs) refer to a group of stable end products produced by the condensation, rearrangement, cleavage, and oxidation modification of free amino groups of macromolecules such as proteins, amino acids, lipids, or nucleic acids with aldehyde groups of reducing sugars under non-enzymatic conditions. The production of AGEs can be inhibited by preventing the formation of early glycation products of reducing sugars.

[0085] AGEs have the property of spontaneous fluorescence, so the content of AGEs can be reflected by fluorescence detection.

[0086] 3. Reagents and consumables

[0087] Main reagents: bovine serum albumin (BSA; Absin, 9048-46-8), PBS buffer (Absin, abs962-500ml), aminoguanidine hydrochloride (AG; MCE, HY-B1041), methylglyoxal (MGO; Aladdin, M109035-250g), 96 ELISA plate (JET BIOFIT, FEP100096), Ketone compounds (1 to 7) were purchased from Nanchang Chennuo Biotechnology Co., Ltd. with a purity of >95%.

[0088]

[0089]

[0090] 4. Experimental equipment

[0091] The experimental equipment is shown in Table 1.

[0092] Table 1 Experimental equipment

[0093] Device Name brand Part Number Constant temperature oscillator Hangzhou Aosheng MB100-4A ELISA reader Thermo VL0L00D0

[0094] 5. Reagent preparation

[0095] a) BSA: Prepare 20 mg / ml BSA in PBS and filter with 0.22 μm filter membrane after complete dissolution;

[0096] b) MGO: The original solution was 40% concentration, which was diluted with PBS to 1% MGO solution;

[0097] c) The positive control compound was aminoguanidine hydrochloride (AG), with final concentrations of 1 mM and 250 μM;

[0098] d) The sample to be tested is dissolved in DMSO to a 20-40 mM stock solution, and the final reaction concentration is 250 μM.

[0099] 6. Experimental process

[0100] a) Add 20 mg / ml BSA solution and 1% MGO solution into a 96-well ELISA plate at a ratio of 1:1;

[0101] b) Add the Yangshen compound / test sample / DMSO to a 96-well ELISA plate, shake and mix, and use a fluorescence ELISA reader to detect the sample fluorescence value RFU under the conditions of an excitation wavelength of 370 nm and an emission wavelength of 440 nm;

[0102] c) Place the 96-well plate in a constant temperature oscillator at 45°C in the dark and incubate for 16 hours;

[0103] d) After 16 hours, the 96-well plate was centrifuged at 1000 rpm for 5 minutes, and the fluorescence value RFU of the sample was detected using a fluorescence microplate reader under the conditions of an excitation wavelength of 370 nm and an emission wavelength of 440 nm.

[0104] 7. Data Analysis

[0105] The AGEs inhibition rate was calculated using the formula for the inhibition rate of the compound on AGEs generation, and the experimental results were expressed as Mean±SD:

[0106]

[0107] Note: The negative control is the fluorescence value of BSA and MGO detected after 16 hours of incubation, the test group is the fluorescence value of the test object detected after 16 hours of incubation, and the blank control is the fluorescence value of BSA and MGO detected at 0 hours.

[0108] T-test is calculated using the T.TEST function of Microsoft Office365 Excel. The specific parameter selection is T.TEST(data-1, data-2, tails=2, type=2), i.e. T.TEST(array-1, array-2, tails=2, type=2).

[0109] 8. Experimental results

[0110] The experimental results are as follows Figure 1 As shown in Tables 2 to 4, in the experiment of inhibiting the formation of AGEs, the AG and 1,7-dihydroxy-2-methoxy-3-(3-methylbut-2-enyl) Compared with 1,7-dihydroxy-2-methoxy-3-(3-methylbut-2-enyl)xanthone (with only one isopentenyl side chain, but the position of its isopentenyl side chain is different from that of the compound claimed in the present invention), the anti-glycation effect is significant; the anti-glycation inhibition rate of Yangshen AG at the same concentration (250μM) is about 17%, and the same concentration (250μM) of 1,7-dihydroxy-2-methoxy-3-(3-methylbut-2-enyl) The anti-glycation inhibition rate of ketone was about 1.5%. Compared with the negative control, the significant T test P values ​​of compounds 1 to 6 were all less than 0.0001, while the significant T test P value of compound 7 was greater than 0.05.

[0111] Table 2 Anti-glycation activity of compounds (AGEs inhibition rate %)

[0112] sample Inhibition rate % (average value) SD T-test vs negative control 1 42.0 1.6 1.38334E-06 2 18.2 0.4 5.22253E-07 3 37.3 0.6 6.48155E-08 4 47.2 1.4 6.24002E-07 5 52.6 1.3 3.14108E-07 6 44.3 0.9 1.4638E-07 7 0.8 0.6 0.101191507 AG 56.1 1.0 8.15937E-08 Negative control (DMSO) 0.0 0.3 1

[0113] Table 3 Significance T test of absorbance values ​​of compound 7 and negative control (DMSO)

[0114]

[0115] Table 4 Significance T test of anti-glycation inhibition rate of compound 7 and negative control (DMSO)

[0116]

[0117]

[0118] Example 2

[0119] Elastase inhibitory activity

[0120] 1. Principle of Elastase Inhibition Assay

[0121] The main characteristics of aging skin are thinning of the epidermis, flattening of the dermal-epidermal junction, and overall degradation of the extracellular matrix (ECM) of the dermis. Elastic fibers play an important role in skin aging. Their degradation and destruction can cause skin elasticity loss, sagging, various wrinkles, and even patches and nodules with lost skin color. Elastase is one of the most important enzymes in the matrix metalloproteinase family. It is mainly synthesized and secreted by fibroblasts. It is a highly selective and specific proteolytic enzyme that can degrade elastin in the skin, leading to a series of aging symptoms such as reduced skin elasticity, sagging, and wrinkles.

[0122] Elastase reacts with the enzyme substrate SucAla3-pNA (N-succinyl-tri-L-alanine 4-nitroanilide) to generate p-nitroaniline (pNA), which is yellow and absorbs at 410nm. The absorbance changes after adding the active substance, and the magnitude of the absorbance change can reflect the inhibition rate of the elastase inhibitor.

[0123] 2. Main materials, reagents and instruments

[0124] Elastase (Porcine pancreas), MedChemExpress; SucAla3-pNA, Tris, Sigma-Aldrich, Co.; Quercetin (purity>98%), Anhui Zesheng Technology Co., Ltd.; Oleanolic acid (purity>98%), Nanjing Yuanzhi Biotechnology Co., Ltd.; ELISA plate, sample tank, Guangzhou Jiete Biofiltration Co., Ltd.; Microplate constant temperature oscillator, Hangzhou Aosheng Instrument Co., Ltd.; XSR205DU / A analytical balance, Mettler-Toledo Instrument Co., Ltd.; Varioskan TM LUX Multi-Mode Microplate Reader, Thermo Fisher Scientific Inc.

[0125] 3. Experimental methods

[0126] 3.1 Solution preparation

[0127] Buffer: 100 mM Tris-HCl, pH 8.0

[0128] SucAla3-pNA: weigh an appropriate amount and dissolve it in Tris-HCl buffer to 1.6 mM

[0129] Elastase: Weigh an appropriate amount and dissolve it in ddH2O to 1 mg / mL, then dilute it with an equal volume of Tris-HCl buffer solution.

[0130] Quercetin: weigh an appropriate amount and dissolve it in DMSO to 3.02 mg / mL

[0131] Oleanolic acid: weigh an appropriate amount and dissolve in DMSO to 5.00 mg / mL

[0132] 3.2 Reaction system and sample testing

[0133] As shown in Table 5, 140 μL Tris-HCl buffer solution, 5 μL sample solutions of different concentrations (test sample solution, or positive control solution, or DMSO as negative control solution) and 5 μL 0.5 mg / mL elastase were added to a 96-well plate, shaken and mixed, and incubated at room temperature for 15 min, then 50 μL 1.6 mM elastase substrate SucAla3-pNA was added, and immediately shaken and mixed, and the absorbance at 0 min was measured at 410 nm using an enzyme reader, followed by incubation at room temperature for 15 min. After 15 min, the absorbance at 15 min was measured at 410 nm using an enzyme reader, and the inhibition rate of the test sample on elastase was calculated by analyzing and processing the data.

[0134] Table 5 Reaction system and sample detection

[0135]

[0136] Note: “-” means not included

[0137] Inhibition rate calculation formula:

[0138] #imgpt22#

[0139] Where:

[0140] A 样品0 The absorption wavelength OD value of the product pNA generated by the sample under test at 410nm at 0min;

[0141] A 样品15 The characteristic absorption wavelength OD value of the product pNA generated when the sample to be tested is measured at 410nm at the 15th minute;

[0142] A 空白0 The characteristic absorption wavelength OD value of the generated product pNA was measured at 410 nm for the negative control group (DMSO) at 0 min;

[0143] A 空白15At 15 minutes, the negative control group (DMSO) was measured at 410 nm for the characteristic absorption wavelength OD value of the generated product pNA.

[0144] 4. Experimental results

[0145] The results of the elastase inhibitory activity assay showed that the inhibition rates (%) of compounds 1 to 6 were between 10% and 50%.

[0146] The embodiments of the present invention are described in detail above. Specific examples are used herein to illustrate the principles and implementation methods of the present invention. The description of the above embodiments is only used to help understand the method of the present invention and its core idea. At the same time, changes or deformations made by those skilled in the art based on the ideas of the present invention, the specific implementation methods and application scope of the present invention, all belong to the scope of protection of the present invention. In summary, the content of this specification should not be understood as limiting the present invention.

Claims

1. Use of a compound of the following formula I or a salt or deuterated product thereof in the preparation of an anti-glycation agent, characterized in that: R1 is hydroxyl or hydrogen, R2 is methoxy or H, R3 is hydroxyl or hydrogen, R4 is methoxy, hydroxyl or H, R5 is isopentenyl, hydroxyl or hydrogen, R6 is methoxy or hydroxyl, and R7 is isopentenyl, methoxy, hydroxyl or hydrogen; wherein R5 and R7 are not isopentenyl at the same time; Wherein, the deuterated substance is a partially deuterated substance and / or a fully deuterated substance.

2. The use according to claim 1, characterized in that The number of isopentenyl groups in the compound of formula I or its salt or deuterated product is 1; Preferably, the number of hydroxyl groups in the compound of formula I or its salt or deuterated product is 2 to 3; Preferably, the number of methoxy groups in the compound of formula I or its salt or deuterated product is 1 to 2; Preferably, the compound of formula I is any one of the following compounds:

3. An anti-glycation composition, characterized in that: The anti-glycation composition comprises a compound of formula I or a salt or deuterated substance thereof; Wherein, the anti-glycation composition is a compound having anti-glycation activity that inhibits the generation of end products of non-enzymatic glycation reactions.

4. Use of the anti-glycation composition according to claim 3 in the preparation of an anti-glycation agent, characterized in that: The anti-glycation composition is a pharmaceutical composition, a composition for skin use, a health product and / or a functional food.

5. The use according to claim 4, characterized in that The pharmaceutical composition further comprises a pharmaceutically acceptable excipient; Preferably, the auxiliary material is selected from one or more of the following: diluent, wetting agent, binder, disintegrant, inclusion agent, flavoring agent, sustained-release agent, retention agent, lubricant, dispersant, plasticizer, opacifier and antioxidant.

6. The use according to claim 4, characterized in that The dosage form of the pharmaceutical composition is a patch, emulsion, tablet, pill, capsule, powder, film, lozenge, granule or oral solution.

7. The use according to claim 4, characterized in that The skin composition further comprises one or more skin care substances for preventing and / or improving skin aging and / or accelerated skin aging, such as carnosine, acetylated hyaluronic acid, adenosine and / or kava extract; Preferably, the skin composition is in the form of an emulsion, an essence, a toner, a cream or a mask.

8. The use according to claim 7, characterized in that The skin aging and / or accelerated skin aging include one or more of the following: dull skin, skin wrinkles, decreased elasticity of skin collagen fibers, and skin pigmentation patches.

9. The use according to claim 7, characterized in that: The skin composition further comprises other auxiliary materials; Preferably, the auxiliary materials are selected from one or more of the following: thickeners, pH regulators, preservatives, antibacterial agents and stabilizers; More preferably, the thickener is selected from one or more of the following: carbomer, sodium polyacrylate, xanthan gum and hydroxyethyl cellulose; More preferably, the pH adjuster is selected from one or more of the following: triethanolamine, aminomethyl propanol, citric acid and lactic acid; More preferably, the preservative is selected from one or more of the following: methylparaben, propylparaben, butylparaben, p-hydroxyacetophenone and phenoxyethanol.

10. The use according to claim 4, characterized in that The health care products and / or functional foods are packaged foods and food raw materials.

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