Preparation of a vinylpyranone compound with anti-inflammatory and antioxidant effects

Abstract

This invention provides a vinylpyranone compound, which has excellent anti-inflammatory and antioxidant activities.

Description

Technical Field

[0001] This invention belongs to the pharmaceutical field, specifically relating to a vinylpyranone compound. Background Technology

[0002] Studies have shown that excessive free radicals can damage tissue cells, subcellular structures, and molecular structures, harming biomolecules such as nucleic acids, lipids, proteins, enzymes, and sugars. As the damage spreads, it causes functional impairment, leading to cardiovascular diseases, cancer, and aging, forming the pathological basis of many diseases. Antioxidant active ingredients can scavenge free radicals and reduce oxidized substances, thus protecting the body.

[0003] Inflammation is a defensive response of living tissue with a vascular system to damaging agents. Most diseases are accompanied by inflammation, which can exacerbate the onset and progression of diseases. Some chronic inflammations can lead to tumors. Therefore, the control and treatment of inflammation are of great importance.

[0004] The search for and development of antioxidants and anti-inflammatory substances that can scavenge oxygen free radicals has become an important research topic in the fields of biology, medicine, chemistry, and pharmaceutics. Summary of the Invention

[0005] This invention provides the following vinylpyranone compounds, their stereoisomers, tautomers, isotope labels, nitrides, solvates, polymorphs, metabolites, esters, pharmaceutically acceptable salts, or prodrugs:

[0006]

[0007] The present invention also provides a pharmaceutical composition comprising one, two or more of the above-mentioned vinylpyranone compounds, their stereoisomers, tautomers, isotope labels, nitrogen oxides, solvates, polymorphs, metabolites, esters, pharmaceutically acceptable salts, and prodrugs.

[0008] According to embodiments of the present invention, the pharmaceutical composition may optionally also contain at least one pharmaceutically acceptable excipient.

[0009] Pharmaceutically acceptable excipients include solvents, propellants, solubilizers, cosolvents, emulsifiers, colorants, binders, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, flow aids, flavoring agents, preservatives, suspending agents, coating materials, flavoring agents, anti-adhesion agents, binding agents, penetration enhancers, pH adjusters, buffers, plasticizers, surfactants, foaming agents, defoamers, thickeners, encapsulating agents, humectants, absorbents, diluents, excipients, flocculants and anti-flocculation agents, filter aids, and release inhibitors.

[0010] According to embodiments of the present invention, the pharmaceutical composition may optionally contain at least one additional active ingredient; specifically, the pharmaceutical composition may also contain one or more active ingredients other than the above-described vinylpyranone compounds, their stereoisomers, tautomers, isotope labels, nitrogen oxides, solvates, polymorphs, metabolites, esters, pharmaceutically acceptable salts, and prodrugs.

[0011] In the pharmaceutical composition described above, the dosage of the aforementioned vinylpyranone compound, its pharmaceutically acceptable salt, solvate, polymorph, metabolite, stereoisomer, tautomer, isotope label, nitride, ester, and prodrug may be a therapeutically effective amount.

[0012] According to embodiments of the present invention, the pharmaceutical compositions of the present invention can be formulated into dosage forms suitable for administration using methods known in the art. According to embodiments of the present invention, the formulations (or pharmaceutical compositions) include: oral formulations and non-oral formulations. According to embodiments of the present invention, the formulations include: powders, granules, capsules, injections, inhalants, tinctures, oral liquids, tablets, lozenges, or drops.

[0013] The present invention also provides the use of the above-mentioned vinylpyranone compounds, their stereoisomers, tautomers, isotope labels, nitrogen oxides, solvates, polymorphs, metabolites, esters, pharmaceutically acceptable salts, prodrugs, or the above-mentioned pharmaceutical compositions in the preparation of anti-inflammatory and antioxidant drugs.

[0014] The present invention also provides an anti-inflammatory and antioxidant method, the method comprising administering to a therapeutically effective amount of the above-mentioned vinylpyranone compound, its stereoisomers, tautomers, isotope labels, nitrogen oxides, solvates, polymorphs, metabolites, esters, pharmaceutically acceptable salts, prodrugs, or one, two, or more of the above-mentioned pharmaceutical compositions to a therapeutic subject.

[0015] Beneficial effects

[0016] This invention provides a vinylpyranone compound, which has excellent anti-inflammatory and antioxidant activities.

[0017] Unless otherwise stated, the definitions of groups and terms recorded in this application specification and claims, including definitions as examples, exemplary definitions, preferred definitions, definitions recorded in tables, and definitions of specific compounds in the examples, can be arbitrarily combined and combined with each other. Such combinations and combinations of group definitions and compound structures should be understood as being within the scope of this application specification and / or claims.

[0018] The stereochemical definitions and rules used in this invention generally follow those of S.P. Parker, Ed., McGraw-Hill Dictionary of Chemical Terms (1984), McGraw-Hill Book Company, New York; and Eliel, E. and Wilen, S., “Stereo chemistry of Organic Compounds,” John Wiley & Sons, Inc., New York, 1994.

[0019] "Stereoisomers" are compounds that have the same chemical structure but whose atoms or groups are arranged differently in space. Stereoisomers include enantiomers, diastereomers, conformational isomers (rotational isomers), geometric isomers (cis / trans) isomers, and hindered isomers, etc.

[0020] "Enantiomers" refer to two non-overlapping but mirror-image isomers of a compound.

[0021] A diastereomer is a stereoisomer that has two or more chiral centers and whose molecules are not mirror images of each other. Diastereomers have different physical properties, such as melting point, boiling point, spectral properties, and reactivity. Mixtures of diastereomers can be separated by high-resolution analytical procedures such as electrophoresis and chromatography, for example, HPLC.

[0022] Any asymmetric atom (e.g., carbon, etc.) in the compounds disclosed in this invention can exist in a racemic or enantiomerically enriched form, such as in (R)-, (S)-, or (R,S)- configurations. In some embodiments, each asymmetric atom has at least 0% enantiomeric excess, at least 60% enantiomeric excess, at least 70% enantiomeric excess, at least 80% enantiomeric excess, at least 90% enantiomeric excess, at least 95% enantiomeric excess, or at least 99% enantiomeric excess in the (R)- or (S)- configuration.

[0023] Any mixture of stereoisomers obtained can be separated into pure or substantially pure geometric isomers, enantiomers, and diastereomers based on differences in the physicochemical properties of the components, for example, by chromatography and / or fractional crystallization.

[0024] In the case of racemic amines, diastereomers are obtained from the mixture by reaction with an optically active resolving agent. Examples of suitable resolving agents are optically active acids, such as tartaric acid in R and S forms, diacetyl tartaric acid, dibenzoyl tartaric acid, mandelic acid, malic acid, lactic acid, suitable N-protected amino acids (e.g., N-benzoylproline or N-benzenesulfonylproline), or various optically active camphorsulfonic acids. Chromatographic enantiomer separation can also be advantageously performed using optically active resolving agents (e.g., dinitrobenzoylphenylglycine immobilized on silica gel, cellulose triacetate or other carbohydrate derivatives, or chiral derivatized isobutylene ester polymers). Suitable eluents for this purpose are aqueous or alcoholic solvent mixtures, such as hexane / isopropanol / acetonitrile.

[0025] The term "tautomer" refers to structural isomers with different energies that can interconvert through a low energy barrier. If tautomerism is possible (e.g., in solution), chemical equilibrium can be achieved for the tautomers. For example, proton tautomers (also called prototropic tautomers) involve interconversions via proton migration, such as keto-enol isomerization and imine-enamine isomerization. Valence tautomers involve interconversions via the rearrangement of some bonding electrons. A specific example of a keto-enol tautomer is the interconversion between pentane-2,4-dione and 4-hydroxypent-3-en-2-one. Another example of tautomerism is phenol-keto tautomerism. A specific example of a phenol-keto tautomer is the interconversion between pyridine-4-ol and pyridine-4(1H)-keto. Unless otherwise stated, all tautomer forms of the compounds of this invention are within the scope of this invention.

[0026] The term "nitrogen oxide" in this invention refers to an N-oxide formed by oxidizing one or more nitrogen atoms when the compound contains several amine functional groups. Specific examples of N-oxides are N-oxides of tertiary amines or N-oxides containing nitrogen atoms in nitrogen-containing heterocyclic nitrogen atoms. The corresponding amines can be treated with oxidizing agents such as hydrogen peroxide or peracids (e.g., peroxycarboxylic acids) to form N-oxides (see Advanced Organic Chemistry, Wiley Interscience, 4th edition, Jerry March, pages). In particular, N-oxides can be prepared using the LWDeady method (Syn. Comm. 1977, 7, 509-514), wherein the amine compound is reacted with m-chloroperoxybenzoic acid (MCPBA), for example in an inert solvent such as dichloromethane.

[0027] The term "isotope label" includes, but is not limited to, isotopes of hydrogen, carbon, nitrogen, oxygen, fluorine, sulfur, and chlorine (e.g., 2 H, 3 H, 13 C, 14 C, 15 N, 18 O, 17 O, 18 F, 35 S and 36 Cl)-labeled compounds of the present invention. Isotope-labeled compounds of the present invention can be used for the determination of the tissue distribution of the compounds, their prodrugs, and metabolites; preferred isotopes for such determinations include 3 H and 14 C. Furthermore, in some cases, substitution with a heavier isotope (e.g., deuterium (2H or D)) can provide increased metabolic stability, which offers therapeutic advantages such as increased in vivo half-life or reduced dose requirements. The isotopically labeled compounds of the present invention can generally be prepared according to the methods described herein by replacing non-isotopically labeled reagents with isotopically labeled reagents.

[0028] The term "pharmaceutically acceptable" refers to molecular entities and compositions that are physiologically tolerable when administered to humans and generally do not produce allergic or similar inappropriate reactions, such as gastrointestinal upset, dizziness, etc.

[0029] The term "carrier" refers to a diluent, excipient, formulation, or matrix that is administered together with the compound. These drug carriers can be sterile liquids, such as water and oils, including those of petroleum, animal, plant, or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil, etc. Aqueous and aqueous solutions, saline solutions, and aqueous glucose and glycerol solutions are preferred as carriers, particularly injectable solutions. Suitable drug carriers are described in EW Martin's "Remington's Pharmaceutical Sciences".

[0030] The term "prodrug" as used in this invention refers to the conversion of a compound into the compound represented by formula (I) in vivo. Such conversion is influenced by the hydrolysis of the prodrug in the blood or its enzymatic conversion into the parent structure in the blood or tissues. The prodrug compounds of this invention can be esters; among existing inventions, esters that can serve as prodrugs include phenyl esters and aliphatic (C) esters. 1-24Esters, acyloxymethyl esters, carbonates, carbamates, and amino acid esters. For example, one compound in this invention contains a hydroxyl group, meaning it can be acylated to yield a prodrug form. Other prodrug forms include phosphate esters, such as those obtained by phosphorylation of a parent compound with a hydroxyl group. For a complete discussion of prodrugs, please refer to the following literature: T. Higuchi and V. Stella, Prodrugs as Novel Delivery Systems, Vol. 14 of the ACSSymposium Series; Edward B. Roche, ed., Bioreversible Carriers in Drug Design, American Pharmaceutical Association and Pergamon Press, 1987; J. Rautio et al., Prodrugs: Design and Clinical Applications, Nature Review Drug Discovery, 2008, 7, 255-270; and SJ Hecker et al., Prodrugs of Phosphates and Phosphonates, Journal of Medicinal Chemistry, 2008, 51, 2328-2345.

[0031] As used in this invention, the term "metabolite" refers to the product obtained in vivo through the metabolism of a specific compound or its salt. A metabolite of a compound can be identified using techniques known in the art, and its activity can be characterized by experimental methods as described in this invention. Such products can be obtained by administering the compound through oxidation, reduction, hydrolysis, acylation, deacylation, esterification, defatting, enzymatic cleavage, etc. Accordingly, this invention includes metabolites of compounds, including metabolites produced by sufficiently exposing the compounds of this invention to mammals for a period of time.

[0032] Pharmaceutically acceptable salts can be, for example, acid addition salts of compounds of the present invention having sufficient basicity and containing a nitrogen atom in the chain or ring, such as acid addition salts formed with inorganic acids including: hydrochloric acid, hydrofluoric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, pyrosulfuric acid, phosphoric acid or nitric acid, or hydrogen sulfate; or acid addition salts formed with organic acids including: formic acid, acetic acid, acetoacetic acid, pyruvic acid, trifluoroacetic acid, propionic acid, butyric acid, hexanoic acid, heptanoic acid, undecanoic acid, lauric acid, benzoic acid, salicylic acid, 2-(4-hydroxybenzoyl)benzoic acid, camphoric acid, cinnamic acid, cyclopentanepropionic acid, and diglucose. Gluconic acid, 3-hydroxy-2-naphtholic acid, nicotinic acid, pyruvic acid, pectinic acid, persulfate, 3-phenylpropionic acid, picric acid, pentyl acid, 2-hydroxyethanesulfonic acid, itaconic acid, aminosulfonic acid, trifluoromethanesulfonic acid, dodecyl sulfuric acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, methanesulfonic acid, 2-naphthalenesulfonic acid, naphthalenedisulfonic acid, camphorsulfonic acid, citric acid, tartaric acid, stearic acid, lactic acid, oxalic acid, malonic acid, succinic acid, malic acid, adipic acid, alginic acid, maleic acid, fumaric acid, D-gluconic acid, mandelic acid, ascorbic acid, glucohepanoic acid, glycerophosphate, aspartic acid, sulfosalicylic acid, hemisulfonic acid or thiocyanate.

[0033] Alternatively, another suitable pharmaceutically acceptable salt of the compounds of the present invention having sufficient acidity is an alkali metal salt (e.g., sodium or potassium salt), an alkaline earth metal salt (e.g., calcium or magnesium salt), an ammonium salt, or a salt formed with an organic base that provides a physiologically acceptable cation, such as a salt formed with sodium ions, potassium ions, N-methylglucosamine, dimethylglucosamine, ethylglucosamine, lysine, dicyclohexylamine, 1,6-hexanediamine, ethanolamine, glucosamine, meglumine, sarcosine, serine, trihydroxymethylaminomethane, aminopropylene glycol, 1-amino-2,3,4-butanetriol. As an example, the pharmaceutically acceptable salts include salts formed by the -COOH group with the following substances: sodium ion, potassium ion, calcium ion, magnesium ion, N-methylglucosamine, dimethylglucosamine, ethylglucosamine, lysine, dicyclohexylamine, 1,6-hexanediamine, ethanolamine, glucosamine, meglumine, sarcosine, serine, trihydroxymethylaminomethane, aminopropylene glycol, and 1-amino-2,3,4-butanetriol.

[0034] In addition, basic nitrogen-containing groups can be quaternized using the following reagents: lower alkyl halides, such as methyl, ethyl, propyl, and butyl chlorides, bromides, and iodides; dialkyl sulfates, such as dimethyl sulfate, diethyl sulfate, dibutyl sulfate, and dipentyl sulfate; long-chain halides, such as decyl, lauryl, myristyl, and stearyl chlorides, bromides, and iodides; and aralkyl halides such as benzyl and phenethyl bromides. As examples, pharmaceutically acceptable salts include hydrochlorides, sulfates, nitrates, hydrogen sulfates, hydrobromides, acetates, oxalates, citrates, methanesulfonates, formates, or meglumine salts.

[0035] Since the compounds of the present invention can have multiple salt-forming sites, the pharmaceutically acceptable salt includes not only the salt formed at one salt-forming site of the compound of the present invention, but also the salt formed at two, three, or all of the salt-forming sites. Therefore, the molar ratio of the compound of formula (I) to the anion of the acid or the cation of the base required for salt formation in the pharmaceutically acceptable salt can vary over a wide range, for example, from 4:1 to 1:4, such as 3:1, 2:1, 1:1, 1:2, 1:3, etc.

[0036] In this invention, "solvent" refers to an association formed by one or more solvent molecules with the compound of this invention. Solvents that form solvates include, but are not limited to, water, isopropanol, ethanol, methanol, dimethyl sulfoxide, ethyl acetate, acetic acid, and aminoethanol. The term "hydrate" refers to an association formed when the solvent molecules are water.

[0037] In this invention, "ester" refers to an ester that is hydrolyzable in vivo, formed from a compound containing a hydroxyl or carboxyl group. Such an ester is, for example, a pharmaceutically acceptable ester that, upon hydrolysis in a human or animal body, produces a parent alcohol or acid. The compounds of formula (I) of this invention contain a carboxyl group and can form hydrolyzable esters in vivo with suitable groups, including, but not limited to, alkyl, arylalkyl, etc.

[0038] As used in this invention, the term "treatment" refers to any disease or condition, and in some embodiments, it means improving the disease or condition (i.e., slowing down or stopping or alleviating the development of the disease or at least one of its clinical symptoms). In other embodiments, "treatment" means alleviating or improving at least one bodily parameter, including bodily parameters that may not be perceived by the patient. In still other embodiments, "treatment" means regulating the disease or condition physically (e.g., stabilizing perceptible symptoms) or physiologically (e.g., stabilizing bodily parameters) or both. In still other embodiments, "treatment" means preventing or delaying the onset, occurrence, or worsening of the disease or condition.

[0039] The term "effective amount" or "therapeutic effective amount" refers to the amount of the compound of this invention sufficient to achieve the intended application (including, but not limited to, the treatment of diseases as defined below). Therapeutic effective amounts may vary depending on factors such as the intended application (in vitro or in vivo), the subject being treated, and the condition of the disease, such as the subject's weight and age, the severity of the disease, and the route of administration, which can be readily determined by those skilled in the art. Specific dosages will vary depending on factors such as the particular compound selected, the administration regimen, whether it is administered in combination with other compounds, the timing of administration, the tissue to which the drug is administered, and the physical delivery system used.

[0040] The pharmaceutical excipients described herein are those widely used in the pharmaceutical manufacturing industry. Excipients primarily serve to provide a safe, stable, and functional pharmaceutical composition, and may also provide methods to ensure that the active ingredient dissolves at a desired rate after administration to a subject, or to promote the effective absorption of the active ingredient after administration to a subject. The pharmaceutical excipients may be inert fillers, or provide a function such as stabilizing the overall pH of the composition or preventing the degradation of the active ingredient. The pharmaceutical excipients may include one or more of the following: binders, suspending agents, emulsifiers, diluents, fillers, granulating agents, adhesives, disintegrants, lubricants, anti-adhesion agents, flow aids, wetting agents, gelling agents, absorption delay agents, dissolution inhibitors, enhancers, adsorbents, buffers, chelating agents, preservatives, colorants, flavoring agents, and sweeteners.

[0041] Substances that can be used as pharmaceutically acceptable excipients include, but are not limited to, ion exchangers, aluminum, aluminum stearate, lecithin, serum proteins such as human serum albumin, buffering substances such as phosphates, glycine, sorbic acid, potassium sorbate, mixtures of partial glycerides of saturated vegetable fatty acids, water, salts or electrolytes such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silicates, magnesium trisilicate, polyvinylpyrrolidone, polyacrylates, waxes, polyethylene-polyoxypropylene-blocking polymers, lanolin, sugars such as lactose, glucose, and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as carboxymethyl cellulose. Sodium thiosulfate, ethyl cellulose and cellulose acetate; gum powder; malt; gelatin; talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols such as propylene glycol and polyethylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffers such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic salts; Ringer's solution; ethanol, phosphate buffer solution, and other non-toxic and suitable lubricants such as sodium lauryl sulfate and magnesium stearate, colorants, release agents, coatings, sweeteners, flavorings and spices, preservatives and antioxidants.

[0042] The pharmaceutical compositions of the present invention can be prepared using any method known to those skilled in the art, based on the disclosure. For example, conventional mixing, dissolving, granulation, emulsification, grinding, encapsulation, embedding, or lyophilization processes.

[0043] The dosage form of the pharmaceutical product of this invention can be selected according to specific circumstances. Pharmaceutical dosage forms often consist of a drug, excipients, and a container / sealing system. One or more excipients (also known as inactive ingredients) can be added to the compounds of this invention to improve or promote the manufacture, stability, administration, and safety of the drug, and to provide a method for obtaining the desired drug release profile. Therefore, the type of excipient added to the drug can be determined by various factors, such as the physical and chemical properties of the drug, the route of administration, and the preparation steps. Pharmaceutical excipients exist in this field and include those listed in various pharmacopoeias. The pharmaceutical compositions of this invention may include one or more physiologically acceptable inactive ingredients that facilitate the processing of the active molecule into a formulation for pharmaceutical use.

[0044] The appropriate formulation depends on the desired route of administration. Routes of administration include intravenous injection, administration via mucosa or nose, and oral administration. For oral administration, compounds can be formulated into liquid or solid dosage forms and presented as immediate-release or controlled-release / sustained-release formulations. Suitable dosage forms for individual oral intake include tablets, pills, sugar-coated pills, hard-shell and soft-shell capsules, liquids, gels, syrups, ointments, suspensions, and emulsions.

[0045] Solid oral dosage forms can be obtained using excipients, including fillers, disintegrants, binders (dry and wet), dissolution retardants, lubricants, flow aids, anti-adhesion agents, cation exchange resins, humectants, antioxidants, preservatives, colorants, and flavoring agents. These excipients can be synthetic or of natural origin. Examples of such excipients include cellulose derivatives, citric acid, dicalcium phosphate, gelatin, magnesium carbonate, magnesium lauryl sulfate / sodium lauryl sulfate, mannitol, polyethylene glycol, polyvinylpyrrolidone, silicates, silica, sodium benzoate, sorbitol, starch, stearic acid or its salts, sugars (i.e., dextrose, sucrose, lactose, etc.), talc, tragacanth gum, hydrogenated vegetable oils, and waxes. Ethanol and water can be used as granulation aids. In some cases, tablets need to be coated with, for example, a taste-masking film, an acid-resistant film, or a delayed-release film. Natural and synthetic polymers are often combined with colorants, sugars, and organic solvents or water to coat tablets, resulting in sugar-coated pills. When capsules are preferred over tablets, their drug powders, suspensions, or solutions can be delivered in compatible hard-shell or soft-shell capsule forms.

[0046] The effective therapeutic dose can first be estimated using various methods well known in the art. The initial dose for animal studies can be based on the effective concentration established in cell culture assays. A suitable dose range for humans can be determined, for example, using data obtained from animal studies and cell culture assays. In some embodiments, the compounds of the present invention can be prepared as oral formulations.

[0047] The appropriate formulation, route of administration, dosage, and dosing interval can be selected based on methods known in the art and taking into account the specific circumstances of the individual.

[0048] In the description of this specification, the references to terms such as "one embodiment," "some embodiments," "example," "specific example," or "some examples," etc., indicate that a specific feature, structure, material, or characteristic described in connection with that embodiment or example is included in at least one embodiment or example of the present invention. In this specification, the illustrative expressions of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more embodiments or examples. Moreover, without contradiction, those skilled in the art can combine and integrate the different embodiments or examples described in this specification, as well as the features of different embodiments or examples.

[0049] Although embodiments of the present invention have been shown and described above, it is understood that the above embodiments are exemplary and should not be construed as limiting the present invention. Those skilled in the art can make changes, modifications, substitutions and variations to the above embodiments within the scope of the present invention. Detailed Implementation

[0050] Preparation Example 1: Preparation of Compound 5a

[0051]

[0052] Step 1:

[0053] Jomeconic acid 1 (5.0 g, 45 mmol), NBS (N-bromosuccinimide) (9.9 g, 56 mmol), and ammonium acetate (4.3 g, 56 mmol) were sequentially added to 50 mL of tetrahydrofuran, and the mixture was stirred and refluxed overnight at 70 °C. The reaction was monitored by TLC until complete. After the reaction mixture cooled to room temperature, insoluble matter was removed by filtration. The filtrate was concentrated to dryness to obtain the crude product. The crude product was purified by silica gel column chromatography to give compound 2 (7.2 g, 85%), a pale yellow solid.

[0054] Step 2:

[0055] Compound 2 (7.0 g, 37 mmol), 18-crown ether-6 (0.96 g, 3.7 mmol), and benzyl bromide (6.8 g, 40 mmol) were sequentially added to 100 mL of dichloromethane, followed by 70 mL of 15% potassium hydroxide aqueous solution, and the mixture was stirred overnight at room temperature. The reaction was monitored by TLC until complete. A suitable amount of water was added to the reaction mixture, and the mixture was extracted three times with dichloromethane. The organic phase was concentrated to dryness to give the crude product. The crude product was purified by silica gel column chromatography to give compound 3 (6.1 g, 59%), a yellow oily liquid.

[0056] Step 3:

[0057] Compound 3 (2.0 g, 7.1 mmol) and 1,2-dimethoxy-3-styrene (1.46 g, 8.9 mmol) were weighed and added to a solution of triethylamine (0.90 g, 8.9 mmol) in 20 mL of N,N-dimethylformamide (DMF). Under N2 protection, palladium acetate (0.16 g, 0.71 mmol) was added to the reaction mixture, and the mixture was stirred overnight at 95 °C. The reaction was monitored by TLC to determine its completeness. A suitable amount of water was added to the reaction mixture, and the mixture was extracted three times with ethyl acetate. The combined organic layers were washed with water and dried over Na2SO4. The organic layers were concentrated to dryness under reduced pressure to give the crude product. The crude product was purified by silica gel column chromatography to give product 4a (0.95 g, 37%).

[0058] Step 4:

[0059] Weigh 0.20 g of 4a and dissolve it in 10 mL of dry dichloromethane. Under N2 protection, add 0.5 mL of BBr3 dropwise and stir at 0 °C. The reaction is monitored by TLC until complete. Quench the reaction with water and concentrate to dryness under reduced pressure to obtain the crude product. Purify the crude product by silica gel chromatography to obtain product 5a (0.078 g, 58%).

[0060] Following the preparation method of compound 5a, compounds 5b, 5d, 5g, 5j, 5k, 5m, and 5n were prepared. The structural formulas and spectral data of compounds 5a, 5b, 5d, 5g, 5j, 5k, 5m, and 5n are shown in Table 1.

[0061] Table 1. Structural formulas and spectral data of compounds.

[0062]

[0063]

[0064] Example 1: Evaluation of in vitro antioxidant activity

[0065] 1. Experimental Materials

[0066] Compounds: Examples: compounds 5a, 5b, 5d, 5g, 5j, 5k, 5m, 5n; Comparative compounds: compound D30 and resveratrol.

[0067] 2 Experimental Methods

[0068] All compounds were screened at a concentration of 2 μM, with three replicates and three independent experiments.

[0069] Add 120 μL of fluorescein (FL, 70 nM, final concentration) and 20 μL of the test compound (2 μM, final concentration) to a black 96-well plate. Incubate at 37 °C for 15 min. Then, rapidly add 60 μL of 2,2'-azobisisobutylamidine dihydrochloride solution (AAPH, 12 mM, final concentration) using a multichannel pipette. Record fluorescence every minute for 120 min, automatically shaking the plate before each read. Use PBS instead of the test compound as a blank control. (Excitation wavelength: 485 nm; emission wavelength: 535 nm)

[0070] ORAC-FL value calculation formula:

[0071] [(AUC S -AUC0) / (AUC T -AUC0)]×[C T / C S ]

[0072] AUC S AUC of the test group; AUC0: AUC of the control group; AUC T Trolox's AUC;

[0073] C T : Concentration of Trolox; C S : Sample concentration

[0074] 3 Experimental Results

[0075] The experimental results are shown in Table 2. Among the tested compounds, compounds 5a, 5b, 5d, and 5g all showed superior antioxidant activity compared with resveratrol and compound D30.

[0076] Table 2 ORAC test results of the compounds

[0077] compound ORAC(eq.) 5a 5.50±0.82 5b 3.65±0.48 5d 5.08±0.74 5g 6.51±0.63 5j 1.85±0.13 5k 1.46±0.10 5m 1.62±0.19 5n 1.30±0.19 D30 3.20±0.43 Resveratrol 3.30±0.18

[0078] Example 2: Evaluation of in vitro anti-inflammatory activity

[0079] 1. Experimental Materials

[0080] Compounds: Examples of compounds 5a, 5b, 5d, 5g, 5j, 5k, 5m, 5n, and compound D30, resveratrol.

[0081] 2 Experimental Methods

[0082] This experiment used mouse microglia (BV-2) as a model. First, the MTT assay was used to test whether the compound had cytotoxicity to BV-2 cells to exclude the interference of the compound on cell growth activity. The drug concentration when the cell viability was >90% was taken. The Griess method was used to determine the NO content in the culture medium and the ELISA method was used to determine the interleukin-6 and tumor necrosis factor content in the culture medium.

[0083] (1) The effect of each compound on the viability of BV-2 cells was detected by MTT assay.

[0084] 100 μL of BV-2 cell suspension (cell density 5 × 10⁶ cells / well) was seeded into 96-well plates. 3 Cells were incubated at 37°C with 5% CO2 for 24 hours before drug administration. Three replicates were set up for background wells, test groups, and blank groups. Different concentrations of sample solution were added to the test groups (DMEM medium was added to the blank groups). After another 24 hours of incubation, 20 μL of MTT solution (5 mg / mL) was added to each well, and the cells were incubated for 4 hours to allow crystals to form. The supernatant was removed, and 150 μL of DMSO was added to each well. The cells were then shaken at low speed for 15 minutes to fully dissolve the crystals. The absorbance at 490 nm was measured using a microplate reader, and cell viability was calculated using the following formula.

[0085] Cell survival rate = (OD) s -OD r ) / (OD0-OD r )×100%

[0086] OD s : Absorbance of the sample group; OD r OD0: Absorbance of the background group; OD0: Absorbance of the blank group.

[0087] (2) Griess method for detecting the effect of each compound on LPS-induced NO release from BV-2 cells

[0088] 100 μL of BV-2 cell suspension (cell density 5 × 10⁶ cells / well) was seeded into 96-well plates. 4 Cells were cultured in 96-well plates (1 cell / well) for 24 hours. LPS (1 μg / mL) was added and incubated for 1 hour, followed by drug administration. After another 24 hours of culture, 50 μL of cell supernatant was transferred to another 96-well plate, and 50 μL each of Griess A and B reagents were added. The plates were incubated in the dark for 10 minutes, and the OD value was measured at 540 nm. The NO concentration was calculated based on a standard curve prepared using NaNO2.

[0089] The experiment was set up with a blank group (added with DMEM medium, no LPS stimulation), an LPS model group (added with DMEM medium, with LPS stimulation), a positive drug group (resveratrol, with LPS stimulation), and a test group (drug, with LPS stimulation), each with 3 replicates, and the experiment was performed 3 times independently.

[0090] (3) ELISA method was used to detect the effects of each compound on LPS-induced release of interleukin-6 and tumor necrosis factor from BV-2 cells.

[0091] The effects of different concentrations of LPS on the release of interleukin-6 and tumor necrosis factor from BV-2 cells were determined according to the experimental procedures of the Thermo Fisher Interleukin-6 and Tumor Necrosis Factor Detection Kit. The IC50 values ​​were calculated by fitting the data using GraphPad Prism8 software. 50 .

[0092] The experiment was set up with a blank group (added with DMEM medium, no LPS stimulation), an LPS model group (added with DMEM medium, with LPS stimulation), a positive drug group (resveratrol, with LPS stimulation), and a test group (drug, with LPS stimulation), each with 3 replicates, and the experiment was performed 3 times independently.

[0093] 3 Experimental Results

[0094] (1) The effect of each compound on the viability of BV-2 cells was detected by MTT assay.

[0095] The MTT assay showed that the cell viability of the compounds was greater than 90% at a concentration of 20 μM. Therefore, the next step of NO determination was carried out under this condition.

[0096] (2) Griess method for detecting the effect of each compound on LPS-induced NO release from BV-2 cells

[0097] The experimental results are shown in Table 3. Compared with resveratrol and compound D30, the tested compounds all showed superior activity in inhibiting the release of NO from BV-2 cells.

[0098] Table 3. Effects of compounds on LPS-induced NO release from BV-2 cells.

[0099] compound <![CDATA[IC that inhibits NO production 50 (μM)]]> 5a 17.96±2.59 5b >20 5d >20 5g 6.57±1.37 5j >20 5k >20 5m >20 5n >20 D30 25.14±1.24 Resveratrol 23.67±1.96

[0100] (3) ELISA method was used to detect the effects of each compound on LPS-induced release of IL-6 and TNF-α from BV-2 cells.

[0101] The experimental results are shown in Table 4. Compared with resveratrol and D30 compound, the tested compounds all showed superior performance in inhibiting the release of interleukin-6 and tumor necrosis factor from BV-2 cells.

[0102] Table 4. Effects of each compound on LPS-induced release of interleukin-6 and tumor necrosis factor from BV-2 cells.

[0103] compound <![CDATA[IC that inhibits IL-6 production 50 (μM)]]> <![CDATA[IC that inhibits TNF-α production 50 (μM)]]> 5a 7.495±1.93 30.14±2.35 5b 5.495±2.14 >20 5d >20 >20 5g 3.065±0.646 20.71±1.49 5j >20 >20 5k >20 >20 5m >20 >20 5n >20 >20 D30 >20 >20 Resveratrol 35.14±1.56 21.97±0.99

[0104] The embodiments of the present invention have been described above. However, the present invention is not limited to the above embodiments. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. The following vinylpyranone compounds or their pharmaceutically acceptable salts:

2. A pharmaceutical composition comprising the vinylpyranone compound of claim 1 or a pharmaceutically acceptable salt thereof.

3. The pharmaceutical composition according to claim 2, characterized in that, The pharmaceutical composition further comprises at least one pharmaceutically acceptable excipient.

4. The pharmaceutical composition according to claim 2, characterized in that, The pharmaceutical composition also contains at least one additional active ingredient.

5. The use of the vinylpyranone compound of claim 1 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of any one of claims 2-4, in the preparation of an antioxidant medicament.

6. The use of the vinylpyranone compound of claim 1 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of any one of claims 2-4, in the preparation of an anti-inflammatory medicament.

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