Application of benzophenone C-glycoside compound in preparation of hypoglycemic drugs

Abstract

The invention relates to the technical field of medicines, in particular to application of a benzophenone C-glycoside compound in preparation of hypoglycemic drugs, and the structural formula of the benzophenone C-glycoside compound is as shown in a formula I: # imgabs0 #. The invention discloses the effect of the benzophenone C-glycoside compound on treating diabetes mellitus for the first time, especially for inhibiting reabsorption of SGLT-2 in glomerulus on metabolic glucose, the pharmacological activity of the benzophenone C-glycoside compound is verified through cell experiments, the benzophenone C-glycoside compound has obvious inhibitory activity on SGLT-2, and the benzophenone C-glycoside compound can be used for treating diabetes mellitus. The traditional Chinese medicine composition can effectively reduce blood sugar of diabetics, has a remarkable curative effect, and has remarkable social value and market value.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to the use of a benzophenone carbonyl glycoside compound in the preparation of a hypoglycemic drug. Background Art

[0002] Diabetes is a group of metabolic diseases characterized by hyperglycemia. Hyperglycemia is caused by defects in insulin secretion, impaired insulin action, or both. Long-term hyperglycemia leads to chronic damage and dysfunction of various tissues, particularly the eyes, kidneys, heart, blood vessels, and nerves. While there are currently numerous clinical frontline treatments with diverse targets, these glucose-lowering drugs are primarily Western medicines, which can be toxic and have significant side effects, and long-term medication can impose a significant financial burden on patients.

[0003] The kidneys contribute approximately 15-55 g / day to gluconeogenesis, meaning that after an overnight fast, the kidneys release 20-25% of glucose into the circulation. The kidneys maintain glucose homeostasis primarily through glomerular glucose reabsorption, which occurs primarily in the proximal renal tubule (PXT). Hyperglycemia in diabetic patients is partially due to glomerular reabsorption of metabolized glucose. Sodium-glucose cotransporter 2 (SGLT-2), located on the luminal side of the first segment of the PXT, has a high affinity for glucose transport. In healthy individuals, SGLT-2 reabsorbs approximately 90% of filtered glucose. Therefore, inhibiting glomerular SGLT-2 reabsorption of metabolized glucose can effectively lower blood glucose in diabetic patients. SGLT-2 inhibitors have become a popular class of glucose-lowering drugs for diabetic patients.

[0004] Benzophenone carbonyl glycosides, as C-glycosides, are characterized by a C-C glycosidic bond at the anomeric carbon. Their unique chemical structure and biological activity have led to diverse pharmaceutical applications, including antioxidant, antiviral, and host-mediated antitumor properties. Compared to their O-glycoside counterparts, C-glycosides exhibit greater stability to hydrolases in vivo. Currently, there are no reports of the benzophenone carbonyl glycosides described herein being used to prevent or treat diabetes. Summary of the Invention

[0005] The purpose of the present invention is to provide a new medical use of benzophenone carbonyl glycoside compounds in terms of hypoglycemic activity.

[0006] In a first aspect, the present invention provides a benzophenone carbonyl glycoside compound having a structure as shown in Formula I or a pharmaceutically acceptable salt, tautomer, or stereoisomer thereof in the preparation of a hypoglycemic drug. Formula I.

[0007] Furthermore, the following applications are also within the scope of protection of the present invention: The benzophenone carbonyl glycoside compound shown in Formula I or its pharmaceutically acceptable salt, tautomer, stereoisomer is used to prepare an SGLT-2 inhibitor.

[0008] The benzophenone carbonyl glycoside compound shown in Formula I or its pharmaceutically acceptable salt, tautomer, stereoisomer can be used alone in the preparation of the drug, or can be used in combination with other components having hypoglycemic activity.

[0009] Furthermore, the drug uses the benzophenone carbonyl glycoside compound shown in Formula I as an active ingredient and is prepared into a pharmaceutically acceptable dosage form using pharmaceutically acceptable excipients.

[0010] Furthermore, the excipients include one or more of solvents, disintegrants, flavoring agents, preservatives, colorants, adhesives, lubricants, diluents and drug carriers.

[0011] It is well understood by those skilled in the art that excipients (e.g., pharmaceutical excipients) include solvents, propellants, solubilizers, cosolvents, emulsifiers, colorants, binders, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, glidants, flavoring agents, preservatives, suspending agents, coating materials, fragrances, anti-adhesive agents, integrities, penetration enhancers, pH regulators, buffers, plasticizers, surfactants, foaming agents, defoaming agents, thickeners, inclusion agents, humectants, absorbents, diluents, flocculants and deflocculating agents, filter aids, and release retardants.

[0012] Among them, the diluent can be one or more of mannitol, sucrose, lactose, sorbitol, xylitol, polyethylene glycol, propylene glycol, vegetable oil, and mineral oil; the disintegrant can be one or more of cross-linked sodium carboxymethyl cellulose, colloidal silicon dioxide, and citric acid; the binder can be one or more of starch slurry, ethanol, water, and povidone alcohol solution; the preservative can be one or more of ethyl paraben, propyl paraben, sorbic acid, potassium sorbate, calcium propionate, sodium dehydroacetate, sodium diacetate, and sodium lactate; the antioxidant can be one or more of ethylenediaminetetraacetic acid, disodium ethylenediaminetetraacetic acid, butylated hydroxytoluene, glycine, inositol, ascorbic acid, hydroxypropyl benzoate ... The flavoring agent may be one or more of aspartame, sucrose, xylitol, steviol glycosides, cyclamate, sorbitol, cocoa, pure vanilla, vanillin, ethyl vanillin, chocolate, malt, and mint; the suspending agent may be one or more of xanthan gum, polyvinyl pyrrolidone, sodium alginate, aluminum stearate, and hydrogenated vegetable oil; the emulsifier may be one or more of alkyl sulfate, soap, dodecylbenzene sulfonate, lactate, sulfosuccinate, monoglyceride sulfonate, phosphate, silicone, and taurate.

[0013] Furthermore, the dosage form includes any one of tablets, capsules, pills, powders, ointments, pills, suspensions, powders, injections, sustained-release preparations, and controlled-release preparations.

[0014] In a second aspect, the present invention provides a hypoglycemic drug comprising a benzophenone carbonyl glycoside compound as shown in Formula I and pharmaceutically acceptable salts, tautomers, stereoisomers and pharmaceutically acceptable excipients.

[0015] The drug is in the form of tablets, capsules, pills, powders, granules, syrups, solutions, emulsions, injections, sprays, aerosols, patches, and can also be in the form of controlled-release or sustained-release dosage forms known in the modern pharmaceutical industry.

[0016] The drug is administered via the gastrointestinal tract and parenteral routes.

[0017] Particularly, the non-gastrointestinal administration route is selected from injection, respiratory tract administration, skin administration, mucosal administration or cavity administration.

[0018] Among them, the parenteral preparation is selected from injections, sprays, aerosols, patches and the like.

[0019] Particularly, the gastrointestinal administration preparation is selected from tablets, capsules, powders, granules, pills, solutions, emulsions or syrups.

[0020] Furthermore, the drug with hypoglycemic activity provided by the present invention can be used in combination with other components with hypoglycemic activity.

[0021] The pharmaceutical composition of the invention contains 0.1-90% (wt) of active ingredients.

[0022] Pharmaceutical compositions can be prepared according to methods known in the art. For this purpose, if necessary, the active ingredient can be combined with one or more solid or liquid pharmaceutical excipients and / or adjuvants to prepare a suitable administration form or dosage form for human use.

[0023] In addition, if necessary, coloring agents, preservatives, perfumes, flavoring agents, sweeteners or other materials may be added to the pharmaceutical preparations.

[0024] Beneficial effects: The present invention discloses for the first time the efficacy of benzophenone glycoside compounds in treating diabetes, especially for inhibiting the reabsorption of metabolic glucose by SGLT-2 in the glomeruli, and verifies the pharmacological activity of benzophenone glycoside compounds through cell experiments. Benzophenone glycoside compounds have obvious inhibitory activity on SGLT-2, can effectively lower the blood sugar of diabetic patients, have significant clinical efficacy, and have significant social value and market value. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 Benzophenone carbonyl glycoside compounds 1 H NMR spectrum; Figure 2 Benzophenone carbonyl glycoside compounds 13 C NMR spectrum; Figure 3 The effect of benzophenone carbonyl glycosides on the inhibitory activity of SGLT-2 in vitro; Figure 4 This is the molecular docking diagram of dibenzophenone carbonyl glycoside compounds and SGLT-2. DETAILED DESCRIPTION

[0026] In order to make those skilled in the art better understand the technical scheme of the present invention, the present invention is described in detail below in conjunction with specific embodiments. The experimental methods for which specific conditions are not specified in the following examples are usually based on conventional conditions or the conditions recommended by the manufacturer. The test materials used in the following examples, unless otherwise specified, are purchased from conventional biochemical reagent stores. Unless otherwise specified, percentages and parts are calculated by weight. Unless otherwise defined, all professional and scientific terms used in the text have the same meaning as those familiar to those skilled in the art. In addition, any method and material similar to or equivalent to the described content can be applied to the present invention. The preferred implementation methods and materials described in the text are only for demonstration purposes.

[0027] The endpoints of the ranges and any values ​​disclosed herein are not limited to the precise ranges or values, and these ranges or values ​​should be understood to include values ​​close to these ranges or values. For numerical ranges, the endpoints of each range, the endpoints of each range and individual point values, and the individual point values ​​can be combined with each other to obtain one or more new numerical ranges, which should be considered to be specifically disclosed herein.

[0028] The technical solutions of the present invention are further described in detail below in conjunction with specific embodiments and drawings. It should be understood that the following embodiments are only used to explain the present invention and are not used to limit the present invention.

[0029] Example 1 Structural Characterization of Benzophenone Carbonyl Glycosides The present inventors obtained benzophenone carbonyl glycoside compounds by multi-step synthesis or extraction from plants according to the aforementioned publications or other public documents; and determined the structure of the compounds by nuclear magnetic resonance spectroscopy. The results of mass spectrometry and nuclear magnetic resonance characterization (such as Figure 1-2 As shown), its chemical structure is determined as shown in Formula I, Formula I.

[0030] like Figure 1-2 As shown, the results of nuclear magnetic resonance spectrometer measurement are as follows: 1 HNMR (400 MHz, CD3OD) spectrum: 1 H-NMR (400 MHz, CD3OD) δ H : 3.36 (1H, m, H-5′′), 3.44 (2H, m, H-3′′, 4′′), 3.71(1H, dd, J = 12.1, 4.8 Hz, H a -6′′), 3.82 (1H, dd, J = 12.1, 2.4 Hz, H b -6′′), 3.88(1H, m, H-2′′), 4.82 (1H, d, J = 6.9 Hz, H-1′′), 6.74 (2H, d, J = 8.8 Hz, H-2′,6′), 7.57 (2H, d, J = 8.7 Hz, H-3′, 5′); 13 CNMR (100 MHz, CD3OD) spectrum: 13 C-NMR (100 MHz, CD3OD) δC : 198.91 (C=O),162.86 (C-2, 6), 161.52 (C-4′), 160.97 (C-3, 5), 133.21 (C-1′), 132.84 (C-2′,6′), 115.47 (C-3′, 5′), 107.13 (C-1), 104.6 (C-4), 82.52 (C-5′′), 79.89 (C-3′′), 76.55 (C-1′′), 73.61 (C-2′′), 71.41 (C-4′′), 62.42 (C-6′′).

[0031] The above spectral data are completely consistent with those reported in the literature. Therefore, it can be confirmed that the dibenzophenone carbon glycoside compound is 2,3,4ʹ,5,6-pentahydroxybenzophenone-4-C-β-D-glucopyranoside.

[0032] Example 2 Investigation of the hypoglycemic activity of 2,3,4ʹ,5,6-pentahydroxybenzophenone-4-C-β-D-glucopyranoside The full-length complementary deoxyribonucleic acid (cDNA) sequence of human SGLT-2 was cloned using standard techniques and stably transfected into Chinese hamster ovary (CHO) cells. Radiolabeled glucose analogs [ 14 C]-methyl-α-D-glucopyranoside ([ 14 C]-AMG, Perkin Elmer) as the transporter substrate.

[0033] Buffer: Prepare the experimental buffer to simulate the low protein conditions of glomerular filtration. The buffer contains: 10 mM hydroxyethylpiperazine ethanesulfonic acid (Hepes), 1.2 mM magnesium chloride (MgCl2), 120 mM sodium chloride (NaCl), 4.7 mM potassium chloride (KCl) and 2.2 mM calcium chloride (CaCl2), pH = 7.4. Use this buffer to prepare 6 μM [ 14 C]-AMG buffer.

[0034] Experimental Methods: CHO cells stably expressing the human SGLT-2 gene were seeded into 96-well cell culture plates containing F12 (1X) medium (Invitrogen) and incubated overnight at 37°C in a 5% CO2 incubator. Two groups were set up: the treatment group received 49 μL of buffer, 1 μL of glycoside compound solution (diluent: DMSO), and 50 μL of 6 μM [ 14C]-AMG buffer; positive group: empagliflozin. Incubate cells at 37 ºC for 1 hour. Stop the uptake reaction by removing the culture medium and stop the incubation by washing the cells with ice-cold lysis buffer. Then add 50 μL of 10% NaOH ice-cold lysis buffer to lyse the cells, transfer the cell lysate into a picoproas-vial, and add 2 mL of Ultima Gold Cocktail. Quantify the radioactivity content in the cells using Tri-carb, and analyze the data using Origin 8.0 software. Adjust the response curve to fit the empirical four-parameter model to determine the half-maximal response concentration of the test compound, i.e., IC 50 The results are as follows Figure 3 shown.

[0035] It can be seen that the positive drug group inhibits SGLT-2 to further absorb glucose IC 50 The IC50 of the benzophenone carbon glycoside compound of the present invention in CHO cells by inhibiting SGLT-2 further uptake of glucose is 4.92 nM, while the IC50 of the benzophenone carbon glycoside compound of the present invention in CHO cells by inhibiting SGLT-2 further uptake of glucose is 9.45 nM.

[0036] Example 3 Investigation of the target site of binding of 2,3,4ʹ,5,6-pentahydroxybenzophenone-4-C-β-D-glucopyranoside to SGLUT-2 Molecular docking experiments were performed using AutoDock Vina software. The software was used (version 1.5.7, The Scripps Research Institute). The crystal structure of the target protein hSGLT2 was obtained from the Protein Data Bank (PDB ID: 8HB0) and pre-processed by removing water molecules, adding polar hydrogen atoms, and assigning Gasteiger charges using AutoDock Tools (ADT). The ligand molecular structure was optimized and converted to PDBQT format. To explore all potential binding sites, a full coverage docking grid region covering the entire protein was defined, and the grid size was set to include the complete protein structure. The exhaustiveness parameter was set to 4, and the maximum number of generated binding modes was 9. The docking simulation was run under default conditions, and the binding affinity was calculated using the Vina scoring function. Post-docking analysis used PyMOL software (version 3.0.3, Schrödinger) to visualize and analyze ligand-protein interactions and binding modes. The experimental results are shown in

[15] . Figure 4 and Table 1.

[0037] Table 1

[0038] Molecular docking results show that the compound's binding free energy to the SGLT-2 protein is comparable to that of the positive control, empagliflozin, indicating that the compound can bind to SGLT-2 via hydrogen bonds, effectively inhibiting the SGLT-2 protein's glucose absorption, thereby achieving a hypoglycemic effect. This is completely consistent with the results of hypoglycemic experiments using CHO cells, further confirming that the target ingredient's hypoglycemic activity is achieved through binding to SGLT-2.

[0039] The benzophenone carbonyl glycoside compound of the present invention is the first reported natural product that achieves hypoglycemic activity by inhibiting the action of SGLT-2. This type of natural product can be used as a lead compound for the development of new hypoglycemic drugs.

[0040] Finally, it should be noted that the above description is only a preferred embodiment of the present invention. Under the guidance of the present invention, those skilled in the art can make various similar expressions without violating the purpose and claims of the present invention. Such changes fall within the scope of protection of the present invention.

Claims

1. Use of a benzophenone carbonyl glycoside compound having a structure as shown in Formula I or a pharmaceutically acceptable salt, tautomer, or stereoisomer thereof in the preparation of a hypoglycemic drug, Formula I.

2. The use according to claim 1, characterized in that For the preparation of SGLT-2 inhibitors.

3. The use according to claim 1, characterized in that The benzophenone carbonyl glycoside compound shown in Formula I or its pharmaceutically acceptable salt, tautomer, stereoisomer can be used alone in the preparation of the drug, or can be used in combination with other components having hypoglycemic activity.

4. The use according to claim 1, characterized in that The drug uses a benzophenone carbonyl glycoside compound with a structure as shown in Formula I as an active ingredient and is prepared into a pharmaceutically acceptable dosage form using pharmaceutically acceptable excipients.

5. The use according to claim 4, characterized in that: The auxiliary materials include one or more of solvents, disintegrants, flavoring agents, preservatives, colorants, binders, lubricants, diluents and drug carriers.

6. The use according to claim 4, characterized in that: The dosage form is an oral dosage form or an injection dosage form, and can also adopt a controlled-release or sustained-release dosage form well known in the modern pharmaceutical industry.

7. A hypoglycemic drug, characterized in that: The invention comprises a benzophenone carbonyl glycoside compound as shown in formula I and pharmaceutically acceptable salts, tautomers, stereoisomers and pharmaceutically acceptable excipients.

8. The hypoglycemic drug according to claim 7, characterized in that Other components with hypoglycemic activity are also included.

9. The hypoglycemic drug according to claim 7 or 8, which is in the form of an oral dosage form or an injectable dosage form, and can also be in the form of a controlled-release or sustained-release dosage form known in the modern pharmaceutical industry.

10. The hypoglycemic drug according to claim 9, characterized in that The auxiliary materials include one or more of solvents, disintegrants, flavoring agents, preservatives, colorants, binders, lubricants, diluents and drug carriers.

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