Medical application of diketone compounds in treating and / or preventing and relieving nerve injury related diseases

Abstract

The invention discloses medical application of diketone compounds in treating and / or preventing and relieving nerve injury related diseases, and relates to application of diketone compounds shown in a general formula (I) or a general formula (II) in preparing medicines for treating and / or preventing and relieving nerve injury related diseases. The compound relieves or recovers damage caused by neuronal excitatory toxicity through at least one of the following mechanisms: blocking allosteric regulation of glutamic acid to acid-sensitive ion channels (ASICs), and inhibiting ASICs or N-methyl-D-aspartic acid receptor (NMDAR) signaling pathways. The invention also relates to application of a medicinal composition, medicinal salt, solvate, hydrate, enantiomer form, diastereoisomer and racemic compound of the compound in preparation of medicines for treating and / or preventing and relieving nerve injury related diseases.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine technology, in particular to diketone compounds, and in particular to the medical use of a class of diketone compounds in treating and / or preventing and alleviating diseases related to nerve damage. Background Art

[0002] Ischemic stroke is the single most disabling disease in my country, with ischemic injury (cerebral infarction) accounting for approximately 70% of cases. Its pathological mechanism involves a complex interaction of glutamate excitotoxicity, oxidative stress, inflammatory cascades, and ion channel dysfunction. Under conditions of ischemia and hypoxia, energy metabolism failure leads to uncontrolled glutamate release from the presynaptic membrane, abnormally elevated glutamate concentrations in the synaptic cleft, and excessive activation of ionotropic glutamate receptors (such as the N-methyl-D-aspartate receptor, NMDAR) triggers intracellular calcium overload, which in turn induces the opening of the mitochondrial permeability transition pore, a burst of reactive oxygen species (ROS), and activation of caspase-dependent apoptotic pathways. Recent studies have found that glutamate significantly amplifies neuronal damage in the acidotic microenvironment by allosterically regulating acid-sensing ion channels (ASICs). ASIC1a isoforms, activated by protons, mediate sustained sodium and calcium influx. Glutamate, by binding to allosteric sites in their extracellular domains (such as the GluR2 acidic pocket), enhances the channel's sensitivity to protons, leading to increased frequency and prolonged opening of the channel. This creates a "glutamate-ASICs" positive feedback loop, ultimately triggering neuronal hyperexcitability, calcium oscillations, and mitochondrial membrane potential collapse. Animal studies have shown that combined inhibition of ASICs and NMDARs can significantly reduce cerebral infarct volume, suggesting the potential advantages of dual-targeted intervention.

[0003] Similar mechanisms are also seen in other neurological injury-related diseases: in Alzheimer's disease (AD), β-amyloid protein (Aβ) deposition can induce abnormal activation of the ASICs / NMDAR pathway; the pathological process of Parkinson's disease (PD) is closely related to oxidative stress and acidosis microenvironment associated with mitochondrial dysfunction; in diseases such as amyotrophic lateral sclerosis (ALS), multiple sclerosis (MS), epilepsy and spinal cord injury (SCI), glutamate excitotoxicity, hyperactivation of the ASICs / NMDAR pathway, free radical accumulation and inflammatory response jointly aggravate neuronal damage; in addition, acidosis-induced ASICs activation and glutamate signaling disorders also exist in the pathological processes of glaucoma, traumatic brain injury (TBI), neuropathic pain and migraine.

[0004] Currently, there are two main categories of neuroprotective agents in clinical use: drugs targeting the glutamate pathway (such as the NMDAR antagonist memantine, the ASICs inhibitor amiloride, and the antioxidant edaravone); and neurorepair metabolic regulators (such as citicoline, butylphthalide, and piracetam). While the former can partially block calcium influx or scavenge free radicals, they suffer from limitations such as a narrow therapeutic window, an inability to intervene in the allosteric enhancement of ASICs by glutamate, and low blood-brain barrier penetration. While the latter can promote membrane repair or improve energy metabolism, they are unable to effectively inhibit the post-stroke glutamate burst release and the synergistic hyperactivation of the ASICs / NMDAR pathway in the acidotic microenvironment, resulting in limited efficacy in intervening on neuronal hyperexcitability and calcium overload. Therefore, developing novel multi-mechanism therapeutic strategies that simultaneously target the dynamic regulation of glutamate and the homeostasis of the acidic microenvironment is of great clinical significance. Summary of the Invention

[0005] In order to overcome the deficiencies of the above-mentioned prior art, the purpose of the present invention is to provide a class of diketone compounds for medical use in the treatment and / or prevention and alleviation of diseases related to nerve damage. Through structural optimization and screening, diketone compounds such as those represented by general formula (I) and general formula (II) were found to block the allosteric regulation of acid-sensing ion channels (ASICs) by glutamate, inhibit ASICs or NMDAR signaling pathways, and alleviate or restore damage caused by neuronal excitotoxicity. They can specifically bind to ASIC1a and NMDAR, significantly reducing nerve cell damage in high-concentration glutamate-induced or oxygen-glucose deprivation (OGD) models, and exhibit free radical scavenging ability. Their multi-target properties can simultaneously intervene in key nodes in the ischemic stroke pathological network (synergistic hyperactivation of the glutamate-ASICs / NMDAR pathway, oxidative stress and inflammatory response), breaking through existing treatment bottlenecks and providing a new strategy for the preparation of drugs for the treatment and / or prevention and alleviation of nerve damage.

[0006] The present invention also relates to the use of pharmaceutical compositions, pharmaceutical salts, solvates, hydrates, enantiomeric forms, diastereoisomers and racemic compounds of such compounds in the preparation of drugs for treating and / or preventing and alleviating diseases related to nerve damage.

[0007] In order to achieve the above object, the technical solution adopted by the present invention is:

[0008] A diketone compound or a pharmaceutically acceptable salt thereof, characterized by being represented by general formula (I) or general formula (II):

[0009]

[0010] In the general formula (I) and formula (II):

[0011] R1 is hydrogen, C1-C4 straight or branched alkyl, C1-C4 acyl, carboxyl, C1-C4 straight or branched alkylcarbonyloxy, or the following groups which are unsubstituted or substituted with 1-2 substituents: C3-C7 cycloalkyl, C5-C7 aromatic ring group, 5-7 membered aromatic heterocyclic group, 5-7 membered heterocyclic group, C2-C7 alkynyl, or C2-C7 alkenyl, wherein each of the heterocyclic group and aromatic heterocyclic group contains 1-3 heteroatoms selected from oxygen, sulfur, and nitrogen;

[0012] The substituents are each independently selected from the following group: halogen, C1-C4 straight or branched alkyl, C1-C4 straight or branched alkoxy, C1-C4 straight or branched ester, cyano, nitro, hydroxy, amino, aminomethyl, dimethylamino, hydroxymethyl, trifluoromethyl, carboxyl, sulfhydryl, C2-C4 acyl, amide, aminosulfonyl, C1-C4 alkyl substituted sulfonyl, or two adjacent substituents together with the carbon atoms to which they are attached form a 5-7 membered ring;

[0013] R2 is a C1-C4 straight-chain or branched alkyl group, a C1-C4 straight-chain or branched alkoxy group, a C1-C4 straight-chain or branched alkylthio group, a C1-C4 straight-chain or branched alkyl-substituted amino group, a hydroxyl group, an amino group, a C3-C7 cycloalkyl group which is unsubstituted or substituted with 1-2 substituents, or a 5-7 membered heterocyclic group;

[0014] The heterocyclic groups each contain 1 to 3 heteroatoms selected from oxygen, sulfur and nitrogen;

[0015] The substituents are independently selected from the following groups: halogen, C1-C4 straight chain or branched alkyl, C1-C4 straight chain or branched alkoxy, C1-C4 straight chain or branched ester, hydroxy, hydroxymethyl, trifluoromethyl, carboxyl, thiol, C2-C4 acyl, amide;

[0016] R3 or R4 is hydrogen, C1-C4 straight or branched alkyl, or both together with the carbon atoms to which they are attached form a 3-7 membered ring;

[0017] n is 0, 1, or 2.

[0018] The diketone compounds represented by general formula (I) or general formula (II) or pharmaceutically acceptable salts thereof have solvates, hydrates, enantiomeric forms, diastereomers and racemic compounds, and are used to prepare (a) allosteric regulatory blockers of glutamate on acid-sensitive ion channels (ASICs), ASICs or N-methyl-D-aspartate receptor (NMDAR) signaling pathway inhibitors; and (b) drugs for treating and / or preventing or alleviating diseases related to nerve damage.

[0019] The nerve damage-related disease is selected from the following group: ischemic brain injury (such as stroke), Alzheimer's disease (AD), Parkinson's disease (PD), amyotrophic lateral sclerosis (ALS), multiple sclerosis (MS), epilepsy, spinal cord injury (SCI), glaucoma, traumatic brain injury (TBI), neuropathic pain, migraine and its complications, or a combination thereof.

[0020] The related diseases caused by nervous system disorders are selected from the following group: Alzheimer's disease, sensory or motor disorders, memory loss, myasthenia gravis, progressive muscular dystrophy, post-poliomyelitis sequelae, cerebral palsy in children, multiple neuritis and its complications, or a combination thereof.

[0021] The compound of the general formula (I) or general formula (II) is selected from the following groups:

[0022]

[0023]

[0024]

[0025]

[0026]

[0027] The compound of the general formula (I) or general formula (II) shown or its pharmaceutically acceptable salt has a solvate, hydrate, enantiomeric form, diastereomer or racemate and is one of the compounds No. 4, 11-14, 16-18, 25, 31, 34, 36, 38, 39, 41, 43 in the above group.

[0028] A pharmaceutical composition comprising: a therapeutically effective amount of a diketone compound represented by general formula (I) or general formula (II) or a pharmaceutically acceptable salt thereof in the form of a solvate, a hydrate, an enantiomer, a diastereomer or a racemate, and an optional pharmaceutically acceptable carrier.

[0029] The method comprises administering a safe and effective amount of a diketone compound represented by general formula (I) or general formula (II), or a pharmaceutically acceptable salt, solvate, hydrate, enantiomer, diastereomer or racemate thereof to a subject in need thereof, wherein the diketone compound represented by general formula (I) or general formula (II) is as described in the first aspect of the present invention.

[0030] The subject is a primate mammal, such as a human.

[0031] Method of drug administration:

[0032] Since the compounds of the present invention can block the allosteric regulatory effect of glutamate on ASICs and directly inhibit the ASICs or NMDAR signaling pathway, the compounds of the present invention and pharmaceutically acceptable salts thereof, as well as pharmaceutical compositions containing the compounds of the present invention as the main active ingredient, which can be used to treat and / or prevent and alleviate diseases related to nerve damage are all included in this patent.

[0033] The pharmaceutical compositions of the present invention comprise a safe and effective amount of a compound of the present invention or a pharmacologically acceptable salt thereof, and a pharmacologically acceptable excipient or carrier. "Safe and effective amount" means an amount of the compound sufficient to significantly improve the condition without causing serious side effects. Typically, the pharmaceutical composition contains 1-2000 mg of the compound of the present invention per dose.

[0034] The "pharmaceutically acceptable salt" of the present invention is a conventional non-toxic salt formed by the reaction of the diketone compound represented by general formula (I) or general formula (II) with an inorganic acid or organic acid. The inorganic acid or organic acid includes hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, aminosulfonic acid, phosphoric acid, citric acid, tartaric acid, lactic acid, pyruvic acid, acetic acid, benzenesulfonic acid, p-toluenesulfonic acid, methanesulfonic acid, naphthalenesulfonic acid, ethanesulfonic acid, naphthalene disulfonic acid, maleic acid, malic acid, malonic acid, fumaric acid, succinic acid, propionic acid, oxalic acid, trifluoroacetic acid, stearic acid, pamoic acid, hydroxymaleic acid, phenylacetic acid, benzoic acid, salicylic acid, glutamic acid, ascorbic acid, p-aminobenzenesulfonic acid, 2-acetoxybenzoic acid and isethionic acid, oleic acid, linoleic acid, Linolenic acid, etc.; or sodium salt, potassium salt, calcium salt, aluminum salt or ammonium salt formed by the compound of general formula (I) or general formula (II) and inorganic base; or methylamine salt, ethylamine salt or ethanolamine salt formed by the compound of general formula (I) or general formula (II) and organic base; or the compound of general formula (I) or general formula (II) forms an ester with lysine, arginine, or ornithine and then forms the corresponding inorganic acid salt with hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid or phosphoric acid, or the corresponding organic acid salt formed with formic acid, acetic acid, picric acid, methanesulfonic acid or ethanesulfonic acid.

[0035] The present invention also relates to a medicament comprising at least one compound of the present invention of general formula (I) or general formula (II), together with one or more pharmacologically acceptable excipients or carriers, and to its use for the above-mentioned purposes. "Pharmaceutically acceptable carrier" refers to one or more compatible solid or liquid fillers or gel substances that are suitable for human use and must be of sufficient purity and sufficiently low toxicity. "Compatibility" as used herein refers to the ability of the components of the composition to be mixed with the compound of the present invention, and with each other, without significantly reducing the compound's efficacy. The pharmaceutical carriers herein include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins such as human albumin, phosphates, glycerol, sorbic acid, potassium sorbate, vegetable oils, partial glyceride mixtures of saturated vegetable fatty acids, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, calcium sulfate, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulosic materials, polyethylene glycol, sodium carboxymethyl cellulose, polyacrylates, beeswax, lanolin, gelatin, talc, solid lubricants, emulsifiers, wetting agents, colorants, flavorings, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0036] There is no particular limitation on the mode of administration of the compound of formula (I) or (II) or pharmaceutical composition of the present invention. Representative modes of administration include, but are not limited to, oral administration, intramuscular or intravenous injection, parenteral administration, pulmonary administration, nasal administration, sublingual administration, lingual administration, buccal administration, rectal administration, transdermal administration, conjunctival administration, topical administration, or administration in the form of an implant.

[0037] Solid dosage forms for oral administration include capsules, tablets (uncoated or coated), pills, powders, and granules. In these solid dosage forms, the active compound of formula (I) or formula (II) is mixed with at least one conventional inert excipient or carrier, such as sodium citrate or dicalcium phosphate, or with the following ingredients: (a) fillers or extenders, such as starch, lactose, sucrose, glucose, mannitol and silicic acid; (b) binders, such as hydroxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose and gum arabic; (c) humectants, such as glycerol; (d) disintegrants, such as agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates, sodium carbonate; (e) solubilizing agents, such as paraffin; (f) absorption accelerators, such as quaternary ammonium compounds; (g) wetting agents, such as cetyl alcohol or glyceryl monostearate; (h) adsorbents, such as kaolin; and (i) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, or mixtures thereof. Capsules, tablets and pills may also contain buffering agents.

[0038] Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells, such as enteric coatings and other materials known in the art. They may contain opacifying agents, and the release of the active compound or compounds in such compositions may be delayed in a certain portion of the digestive tract. Examples of useful encapsulating components are polymeric substances and waxes; if desired, the active compound may be microencapsulated with one or more of the above-mentioned excipients.

[0039] Liquid dosage forms for oral administration include pharmaceutically acceptable solutions, emulsions, suspensions, syrups or tinctures. In addition to the active compound, the liquid dosage form may contain an inert diluent conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, such as ethanol, isopropyl alcohol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butylene glycol, dimethylformamide, dimethyl sulfoxide, and cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or mixtures thereof.

[0040] In addition to these inert diluents, the composition may also contain adjuvants such as wetting agents, emulsifying and suspending agents, sweeteners, flavoring agents and perfumes. In addition to the active compound, the suspension may contain suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these substances.

[0041] Compositions for parenteral injection may include physiologically acceptable sterile aqueous or anhydrous ethanol, polyol solutions, dispersions, suspensions, emulsions, lyophilized products and sterile powders, and sterile powders for reconstitution into sterile injectable solutions or dispersions, suitable aqueous and non-aqueous carriers and diluents.

[0042] Dosage forms suitable for other routes of administration include nasal drops / solutions, sprays; tablets or capsules for lingual, sublingual or buccal administration, suppositories, preparations for the ears and eyes, vaginal capsules, aqueous suspensions (lotions, shaken mixtures), lipophilic suspensions, ointments, creams, emulsions, pastes, dusting powders or implants.

[0043] The active ingredient of general formula (I) or general formula (II) can be converted into the above-mentioned administration forms by methods known per se, which can be achieved by using suitable inert non-toxic pharmaceutical excipients, including carriers such as microcrystalline cellulose, solvents such as liquid polyethylene glycol, emulsifiers such as sodium lauryl sulfate, dispersants such as polyvinyl pyrrolidone, synthetic and natural biopolymers, stabilizers such as antioxidants, colorants such as inorganic pigments or flavoring agents and / or taste-masking agents. Where appropriate, the active ingredient can be present in one or more of the above-mentioned carriers in the form of microencapsulation.

[0044] In addition to the compound of general formula (I) or general formula (II) of the present invention, the above-mentioned pharmaceutical preparations may also contain other pharmaceutically active ingredients.

[0045] When using a pharmaceutical composition, a safe and effective amount of the compound of the present invention is applied to a mammal (such as a human) in need of treatment, wherein the administered dosage is a pharmaceutically effective dosage. For a person weighing 60 kg, the daily dosage is generally 1 to 2000 mg; of course, the specific dosage should also be considered in light of factors such as the route of administration and the patient's health condition.

[0046] The advantages of the present invention mainly include: the present invention provides a diketone compound with a structure represented by general formula (I) or general formula (II), which can block the allosteric regulation of glutamate on ASICs, inhibit ASICs or NMDAR signaling pathways, and has the potential to prepare drugs for treating and / or preventing and alleviating diseases related to nerve damage. DETAILED DESCRIPTION

[0047] The present invention is further described below with reference to the following embodiments. Within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features described in detail below can be combined with each other to form a new technical solution. The present invention is not limited to the following embodiments.

[0048] English abbreviations and their full Chinese names

[0049]

[0050] the term:

[0051] The term "aromatic ring group" refers to a group having aromaticity, such as but not limited to: phenyl, naphthyl.

[0052] The term "5- to 6-membered aromatic heterocyclic group" refers to a 5- or 6-membered aromatic group having one or more heteroatoms selected from nitrogen, oxygen or sulfur, which can be selected from the following group: pyridyl, pyrimidinyl, thiazolyl, isothiazolyl, furyl, thienyl, pyrrolyl.

[0053] The term "1 to 2 azaphenyl groups" refers to a 6-membered aromatic group having 1 or 2 nitrogen heteroatoms, which can be selected from the following group: pyridyl, pyrimidinyl, pyridazinyl, pyrazinyl.

[0054] The term "5- to 6-membered heterocyclic group" refers to a 5-, 6-, or 7-membered cyclic group having one or more heteroatoms selected from nitrogen, oxygen, or sulfur, which can be selected from the following groups: tetrahydrofuranyl, piperidinyl, piperazinyl, morpholinyl, pyrrolidinyl, tetrahydrothiazolyl, dioxane, or similar groups.

[0055] The term "halogen" refers to fluorine, chlorine, bromine, or iodine.

[0056] The term "halogenated" refers to fluoro, chloro, brominated, iodinated.

[0057] The term "C1-C4 alkyl" refers to a straight or branched alkyl group having 1 to 4 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, and tert-butyl.

[0058] The term "C1-C4 alkoxy" refers to a straight or branched alkoxy group having 1 to 4 carbon atoms, such as methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec-butoxy, tert-butoxy, or the like.

[0059] The term "C2-C6 ester group" refers to a ROC(=O)- group having 2 to 6 carbon atoms, such as -COOCH3, -COOC2H5, -COOC3H7, -COOC4H9, or the like.

[0060] In this document, unless otherwise specified, the term "substituted" means that one or more hydrogen atoms on the group are replaced by a substituent selected from the following groups: C1-C4 alkyl, C1-C4 alkoxy, halogen, hydroxy, carboxyl, C2-C4 acyl, C2-C6 ester, amino, phenyl; the phenyl group includes unsubstituted phenyl or substituted phenyl with 1-3 substituents selected from the group consisting of halogen, C1-C4 alkyl, cyano, hydroxy, nitro, C3-C6 cycloalkyl, C1-C4 alkoxy, and amino.

[0061] Unless otherwise indicated, the structural formulas described herein are intended to include all isomeric forms (e.g., enantiomers, diastereomers, and geometric isomers (or conformers)): for example, R and S configurations containing asymmetric centers, (Z) and (E) isomers of double bonds, and (Z) and (E) conformers. Therefore, individual stereochemical isomers of the compounds of the present invention, or mixtures of their enantiomers, diastereomers, or geometric isomers (or conformers), are within the scope of the present invention.

[0062] Example 1: Synthesis of Compound 1 (9-(4-hydroxy-3-methoxyphenyl)-3,3,6,6-tetramethyl-3,4,6,7,9,10-hexahydroacridine-1,8(2H,5H)-dione)

[0063] Step 1. Synthesis of catalyst (Cata-1)

[0064]

[0065] To a 250mL round-bottom flask, add 11.22g (100mmol) of triethylenediamine and dissolve it in 70mL of 75% ethanol. Stir in an ice-water bath. Add 24.43g of 1,3-propane sultone (200mmol, 5mL of ethanol to prevent solidification) dropwise into a constant-pressure dropping funnel over 15 minutes. Stir for 12 hours. Add 75mL of distilled water and stir at room temperature for another 4 hours. Add 20.2g of concentrated sulfuric acid (202mmol) slowly dropwise over 15 minutes. Stir at room temperature for 30 minutes, then at 60°C for 10 hours to obtain the desired liquid catalyst (Cata-1).

[0066] Step 2. Synthesis of Compound 1 (9-(4-hydroxy-3-methoxyphenyl)-3,3,6,6-tetramethyl-3,4,6,7,9,10-hexahydroacridine-1,8(2H,5H)-dione)

[0067]

[0068] To a 100mL round-bottom flask, add 0.76g (5mmol) of 3-methoxy-4-hydroxybenzaldehyde (vanillin), 1.40g (10mol) of 5,5-dimethyl-1,3-cyclohexanedione (dalmethylenedione), and 0.58g (7.5mol) of ammonium acetate. Add 20mL of 95% ethanol and 0.1mL of Cata-1 catalyst. Reflux in an oil bath for 8 hours to precipitate. Stop heating and stirring, cool naturally to below 40°C, filter, and wash with 10mL of anhydrous ethanol, then 10mL of methyl tert-butyl ether. Drain to dryness to obtain a white solid. Dry in a vacuum at 75°C and weigh 1.34g, for a yield of 68%.

[0069] 1 H NMR(400MHz,DMSO)δ9.10(d,J=7.5Hz,1H),8.60(s,1H),6.67(d,J=8.0Hz,2H),6.51(s,1H),4.6 7(s,1H),3.57(d,J=69.3Hz,3H),2.47-2.22(m,4H),2.22-1.83(m,4H),0.94(d,J=47.3Hz,12H). 13 C NMR(101MHz,DMSO)δ194.42(2C),148.94(2C),146.67,144.31,138.48,119.74,114.73,11 2.19,111.73(2C),55.47,50.29(2C),32.09(2C),31.85(2C),29.15,26.36(4C).HRMS(ESI + )m / z calcd for C 24 H30 NO4 + 396.2175found396.2169Da.

[0070] Example 2: Synthesis of Compound 3 (4-(4-hydroxy-3-methoxyphenyl)-2,6-dimethyl-1,4-dihydropyridine-3,5-dicarboxylic acid diethyl ester)

[0071]

[0072] To a 50 mL round-bottom flask, add 0.76 g (5 mmol) of 3-methoxy-4-hydroxybenzaldehyde (vanillin), 1.30 g (10.0 mmol) of ethyl acetoacetate, and 0.58 g (7.5 mmol) of ammonium acetate. Add 10 mL of acetonitrile, then add 0.09 g (0.5 mmol) of p-toluenesulfonic acid (p-TSA). Heat the mixture under reflux in an oil bath and stir for 12 hours. After cooling to room temperature, the reaction mixture was directly mixed and purified by column chromatography (DCM / MeOH = 30:1, 0.1% aqueous ammonia) to obtain a light yellow solid weighing 1.69 g, with a yield of 90%.

[0073] 1 H NMR(400MHz, CDCl3)δ7.26(s,1H),6.96-6.69(m,1H),5.58(s,1H),4.34-4.00( m,4H),3.86(d,J=18.0Hz,3H),2.46-2.14(m,6H),1.27(dt,J=22.2,7.1Hz,6H). 13 C NMR(101MHz,DMSO)δ167.68(2C),145.80,143.91,143.46(2C),140.06,120.51,113. 85,110.92,104.40(2C),59.70(2C),55.77,39.16,19.64(2C),14.33(2C).HRMS(ESI - )m / zcalcd for C 20 H 24 NO6 - 374.1604 found 374.1603Da.

[0074] Example 3: Synthesis of Compound 5 (4-ethyl-2,6-dimethyl-1,4-dihydropyridine-3,5-dicarboxylic acid dimethyl ester)

[0075]

[0076] To a 50 mL round-bottom flask, 0.35 g (6 mmol) of propanal, 1.30 g (10.0 mmol) of ethyl acetoacetate, and 0.58 g (7.5 mmol) of ammonium acetate were added, along with 8 mL of acetonitrile. p-Toluenesulfonic acid (p-TSA) (0.09 g, 0.5 mmol) was then added. The mixture was heated to reflux in an oil bath and stirred for 10 h to precipitate. Heating and stirring were discontinued, the mixture was cooled to room temperature, filtered, and washed with 10 mL of anhydrous ethanol, then 10 mL of methyl tert-butyl ether, and dried to obtain a pale yellow solid. TLC (DCM / MeOH = 20:1, 0.1% ammonia water) confirmed the product to be pure. The product was dried under vacuum at 75°C, weighing 0.90 g, for a yield of 64%.

[0077] 1 H NMR (400MHz, CDCl3) δ4.34-4.06(m,4H),3.91(t,J=5.6Hz,1H),2.29(s,6H),1.42-1.32(m,2H),1.29(t,J=7.1Hz,6H),0.75(t,J=7.5Hz,3H). 13 C NMR(101MHz, CDCl3)δ168.19(2C),144.73(2C),102.81(2C),59.56(2C),34.10,29.29,19.48(2C),14.42(2C),9.25.HRMS(ESI + )m / z calcd for C 15 H 24 NO4 + 282.1705found 282.1687Da.

[0078] Example 4: Synthesis of Compound 11 (9-(3-hydroxy-5-(hydroxymethyl)-2-methylpyridin-4-yl)-3,4,6,7,9,10-hexahydroacridine-1,8(2H,5H)-dione)

[0079]

[0080] To a 100 mL round-bottom flask, add 0.84 g (5 mmol) of 3-hydroxy-5-(hydroxymethyl)-2-methylisonicotinamide aldehyde (pyridoxal), 1.12 g (10 mol) of 1,3-cyclohexanedione, and 0.58 g (7.5 mol) of ammonium acetate. Add 20 mL of 95% ethanol and 0.1 mL of Cata-1 catalyst. Reflux in an oil bath for 15 hours to precipitate. Discontinue heating and stirring, cool naturally to 20°C, filter, and wash with 10 mL of anhydrous ethanol, then 10 mL of methyl tert-butyl ether. Drain dry to obtain a pale yellow solid. Dry in a vacuum at 75°C and weigh 0.87 g, for a yield of 49%.

[0081] 1 H NMR(400MHz,DMSO)δ9.86(s,1H),7.83(s,1H),5.76(s,1H),5.01(s,2H),4 .39(s,1H),2.50(s,4H),2.29(s,3H),2.16(s,4H),1.81(d,J=35.2Hz,4H). 13 C NMR(101MHz, CDCl3)δ195.78(2C),189.33,153.54(2C),153.40,147.12,139.90,137.72 ,133.74,109.74(2C),59.56,48.44(2C),39.31,21.35(2C),20.44(2C),19.50.HRMS(ESI + )m / zcalcd forC 20 H 23 N2O4 + 355.1658found 355.1653Da.

[0082] Example 5: Synthesis of Compound 13 (2-(1,8-dioxo-1,2,3,4,5,6,7,8,9,10-decahydroacridin-9-yl)benzenesulfonic acid)

[0083]

[0084] To a 100 mL round-bottom flask, add 0.93 g (5 mmol) of 2-formylbenzenesulfonic acid, 1.12 g (10 mol) of 1,3-cyclohexanedione, and 0.58 g (7.5 mol) of ammonium acetate. Add 20 mL of 95% ethanol and 0.1 mL of Cata-1 catalyst. Reflux in an oil bath for 10 hours to precipitate. Discontinue heating and stirring, cool naturally to below 40°C, filter, and wash with 10 mL of anhydrous ethanol, then 10 mL of methyl tert-butyl ether. Drain dry to obtain a white solid. Dry under vacuum at 75°C and weigh 1.72 g, for a yield of 92%.

[0085] 1 H NMR (400MHz, DMSO) δ9.15(s,1H),7.71(d,J=7.6Hz,1H),7.33(d,J=7.5Hz,1H),7.11(dd,J=15.2,7.6 Hz,3H),6.98(t,J=7.2Hz,1H),5.72(s,1H),2.37(dd,J=26.6,19.6Hz,4H),2.08(s,4H),1.79(s,4H). 13C NMR(101MHz,DMSO)δ194.79(2C),151.68(2C),145.41,142.80,133.06,128.64,1 27.68,124.63,113.20(2C),37.00(2C),33.89,26.66(2C),20.41(2C).HRMS(ESI + )m / z calcd for C 25 H 24 NO8S2 - 530.0943 found 530.1952Da.

[0086] Example 6: Synthesis of Compound 17 (9-(4-(diethylamino)-2-hydroxyphenyl)-3,3,6,6-tetramethyl-3,4,6,7,9,10-hexahydroacridine-1,8(2H,5H)-dione)

[0087]

[0088] To a 100mL round-bottom flask, add 0.97g (5mmol) of 4-(diethylamino)-2-hydroxybenzaldehyde, 1.40g (10mol) of 5,5-dimethyl-1,3-cyclohexanedione (Damione), and 0.58g (7.5mol) of ammonium acetate. Add 20mL of 95% ethanol and 0.1mL of Cata-1 catalyst. Reflux in an oil bath for 7h to precipitate. Stop heating and stirring, cool naturally to 20°C, filter, and wash with 10mL of anhydrous ethanol, then 10mL of methyl tert-butyl ether. Drain to dryness to obtain an off-white solid. Dry in a vacuum at 75°C and weigh 0.61g, for a yield of 29%.

[0089] 1 H NMR (400MHz, CDCl3) δ6.80 (d, J = 8.1Hz, 1H), 6.45-6.19 (m, 2H), 4.57 (s, 1H), 3.29 (d, J = 6.9Hz, 4H), 2.57 (d, J = 17.3Hz, 1H), 2. 41(d,J=17.3Hz,1H),2.32-2.12(m,4H),2.02(d,J=14.7Hz,2H),1.13(t,J=6.9Hz,6H),1.09(s,3H),1.04(s,3H),0.96(s,6H). 13CNMR(101MHz, CDCl3)δ194.18(2C),157.54,147.37(2C),128.60,121.16,113.89,111.92(3C),108.34,9 8.51,50.85(2C),44.31(2C),41.68(2C),32.11(2C),31.60,27.60(2C),28.99(2C),12.67(2C).HRMS(ESI + )m / z calcd for C 27 H 37 N2O3 + 437.2804found437.2799Da.

[0090] Example 7: Synthesis of Compound 31 (9-acetyl-3,3,6,6-tetramethyl-3,4,6,7,9,10-hexahydroacridine-1,8(2H,5H)-dione)

[0091]

[0092] To a 100mL round-bottom flask, add 0.43g (6mmol) of methylglyoxal, 1.40g (10mol) of 5,5-dimethyl-1,3-cyclohexanedione (Damione), and 0.58g (7.5mol) of ammonium acetate. 15mL of 95% ethanol was added, followed by 0.1mL of Cata-1 catalyst. The mixture was heated and refluxed in an oil bath for 18h to precipitate. Heating and stirring were stopped, and the mixture was naturally cooled to 20°C. Filtered, washed with 10mL of anhydrous ethanol, then 10mL of methyl tert-butyl ether, and dried to obtain a white solid. The solid was vacuum dried at 75°C and weighed 1.12g, for a yield of 71%.

[0093] 1 H NMR (400MHz, DMSO) δ10.86(s,1H),9.52(s,1H),2.58(q,J=17.2Hz,2H),2.36(q,J=17. 2Hz, 2H), 2.05 (dd, J=32.7, 17.0Hz, 4H), 1.84 (s, 3H), 1.12 (s, 3H), 1.08-0.90 (m, 9H). 13 C NMR(101MHz,DMSO)δ190.97(2C),168.82,140.20,126.27,117.44,109.19,108.44,52 .49(2C),36.31,35.08(2C),31.54(2C),28.28(2C),27.84(2C),11.25(2C).HRMS(ESI + )m / z calcd for C19 H 26 NO3 + 316.1913found 316.1907Da.

[0094] Example 8: Synthesis of Compound 40 (3,3,6,6-tetramethyl-3,4,6,7,9,10-hexahydroacridine-1,8(2H,5H)-dione)

[0095]

[0096] To a 100mL round-bottom flask, add 0.81g (10mmol) of 37% formaldehyde solution, 1.40g (10mol) of 5,5-dimethyl-1,3-cyclohexanedione (Damione), and 0.58g (7.5mol) of ammonium acetate. 15mL of 95% ethanol and 0.1mL of Cata-1 catalyst were added. The mixture was heated and refluxed in an oil bath for 15h to precipitate. Heating and stirring were stopped, and the mixture was naturally cooled to 20°C. Filtered, washed with 10mL of anhydrous ethanol, then 10mL of methyl tert-butyl ether, and dried to obtain a white solid. Dry in a vacuum at 75°C and weigh 1.06g, for a yield of 77%.

[0097] 1 H NMR (400MHz, DMSO) δ8.86(s,1H),2.83(s,2H),2.24(s,4H),2.13(s,4H),0.99(s,12H). 13 C NMR(101MHz,DMSO)δ195.36(2C),150.33(2C),107.20(2C),49.94(2C),31.94(2C),27.83(4C),18.30.HRMS(ESI + )m / z calcd for C 17 H 24 NO2 + 274.1807found274.1802Da.

[0098] Example 9: Synthesis of Compound 41 (4-(4-(3,3,6,6-tetramethyl-1,8-dioxo-1,2,3,4,5,6,7,8,9,10-decahydroacridin-9-yl)benzamido)benzoic acid ethyl ester)

[0099] Synthesis route:

[0100]

[0101] Step 1. Synthesis of Compound 22 (4-(3,3,6,6-tetramethyl-1,8-dioxo-1,2,3,4,5,6,7,8,9,10-decahydroacridin-9-yl)benzoic acid)

[0102] To a 100 mL round-bottom flask, 0.75 g (5 mmol) of p-formylbenzoic acid, 1.40 g (10 mol) of 5,5-dimethyl-1,3-cyclohexanedione (Damione), and 0.58 g (7.5 mol) of ammonium acetate were added. 15 mL of 95% ethanol was then added, followed by 0.1 mL of Cata-1 catalyst. The mixture was heated and refluxed in an oil bath for 16 h to precipitate. Heating and stirring were stopped, and the mixture was naturally cooled to below 40°C. The mixture was filtered, washed with 10 mL of anhydrous ethanol, then 10 mL of methyl tert-butyl ether, and dried to obtain a light yellow solid. The solid was dried in vacuo at 75°C and weighed 1.77 g, compound 22, with a yield of 90%.

[0103] 1 H NMR (400MHz, DMSO) δ12.67(s,1H),9.36(s,1H),7.75(d,J=8.2Hz,2H),7.27(d,J=8.3Hz,2H),4.87(s,1H),2.47(d,J=17 .2Hz,2H),2.34(d,J=17.0Hz,2H),2.18(d,J=16.1Hz,2H),1.98(d,J=16.1Hz,2H),0.99(d,J=19.1Hz,6H),0.85(s,6H). 13 CNMR(101MHz,DMSO)δ194.30(2C),167.25,151.97,149.61(2C),128.80(2C),128.09,127. 80(2C),110.89(2C),50.14(2C),33.36(2C),32.11(3C),29.04(2C),26.39(2C).HRMS(ESI + )m / z calcd for C 24 H 28 NO4 + 394.2018found 394.2014Da.

[0104] Step 2. Synthesis of Compound 41 (ethyl 4-(4-(3,3,6,6-tetramethyl-1,8-dioxo-1,2,3,4,5,6,7,8,9,10-decahydroacridin-9-yl)benzamido)benzoate)

[0105] To a 50 mL round-bottom flask was added 220.78 g (2 mmol) of the compound obtained in the previous step, 10 mL of DMF, 0.34 g (2 mmol) of benzocaine, 0.34 g (2.5 mmol) of HOBT, 0.48 g (2.5 mmol) of EDCI, and 0.51 g (5 mmol) of triethylamine. The mixture was stirred at room temperature for 12 hours. 20 mL of distilled water was added to the reaction solution, and the mixture was extracted with 2 x 30 mL of CH2Cl2. The organic phase was washed with 0.1 N hydrochloric acid (10 mL), saturated aqueous sodium bicarbonate (10 mL), and saturated brine (10 mL). The organic phase was dried over anhydrous Na2SO4, filtered, rotary evaporated, and purified by column chromatography (CH2Cl2 / MeOH = 30 / 1, 0.1% ammonia) to obtain a white solid weighing 0.78 g, with a yield of 78%.

[0106] 1 H NMR (400MHz, CDCl3) δ8.14(d,J=7.4Hz,2H),8.08(d,J=8.7Hz,1H),7.61(d,J=7.4Hz,2H),7.59-7.52(m,1H),7.45(d,J=5.4 Hz,2H),6.56(s,1H),5.19(s,1H),2.44(d,J=16.7Hz,2H),2.36-1.92(m,8H),1.36-1.16(m,3H),1.10(s,7H),0.96(s,7H). 13 C NMR(101MHz, CDCl3)δ195.22(2C),162.83,154.84,148.11(2C),143.53,130.69(2C),129.19(2C),128.84( 2C),124.93,122.05,120.38,112.82(2C),108.54(2C),50.62,41.18,34.79,32.72,29.49,27.11.HRMS(ESI + )m / z calcd for C 33 H 37 N2O5 + 541.2702found 541.2711Da.

[0107] Example 10: Synthesis of Compound 43 (9-(3-(diethylamino)methyl)-4-hydroxyphenyl)-3,3,6,6-tetramethyl-3,4,6,7,9,10-hexahydroacridine-1,8(2H,5H)-dione)

[0108] Synthesis route:

[0109]

[0110] Step 1. Synthesis of the intermediate (3-((diethylamino)methyl)-4-hydroxybenzaldehyde)

[0111] To a 100-mL round-bottom flask were added 3.61 g (30 mmol) of p-hydroxybenzaldehyde, 30 mL of ethanol, and 1.28 g (43 mmol) of paraformaldehyde. Then, 3.3 g (45 mmol) of diethylamine and 0.3 mL of concentrated hydrochloric acid were added, and the mixture was heated under reflux for 8 h. The solvent was concentrated under reduced pressure, 10 mL of water was added, and the pH was adjusted to approximately 7 with saturated aqueous sodium bicarbonate solution. The mixture was then extracted with 2 x 30 mL of ethyl acetate. The organic phase was washed with 20 mL of saturated brine, dried over Na2SO4, filtered, rotary evaporated, and purified by column chromatography (CH2Cl2 / MeOH = 20 / 1, 0.1% aqueous ammonia) to obtain a light yellow solid weighing 4.1 g, for a yield of 66%.

[0112] 1 H NMR (CDCl3, 400MHz) δ (ppm) 12.22–11.55 (m, 1H), 9.78 (s, 1H), 7.67 (dd, J = 8.3, 1.9Hz, 1H), 7.53 ( d,J=1.9Hz,1H),6.86(d,J=8.3Hz,1H),3.84(s,2H),2.64(q,J=7.2Hz,4H),1.12(t,J=7.2Hz,6H).

[0113] Step 2. Synthesis of Compound 43 (9-(3-(diethylamino)methyl)-4-hydroxyphenyl)-3,3,6,6-tetramethyl-3,4,6,7,9,10-hexahydroacridine-1,8(2H,5H)-dione)

[0114] To a 100 mL round-bottom flask, add 1.04 g (5 mmol) of the intermediate (3-((diethylamino)methyl)-4-hydroxybenzaldehyde) obtained in the previous step, 1.40 g (10 mol) of 5,5-dimethyl-1,3-cyclohexanedione (Damione), and 0.58 g (7.5 mol) of ammonium acetate. Add 20 mL of 95% ethanol and 0.1 mL of the catalyst Cata-1. Reflux in an oil bath for 10 h to precipitate. Stop heating and stirring, cool naturally to 20°C, filter, wash with 10 mL of anhydrous ethanol, then with 10 mL of methyl tert-butyl ether, and drain to obtain a white solid. Dry in a vacuum at 75°C and weigh 1.55 g, for a yield of 69%.

[0115] 1H NMR (400MHz, DMSO) δ7.15(d,J=1.8Hz,1H),7.00(dd,J=8.4,2.1Hz,1H),6.75(d,J=8.4Hz,1H),4.72(s,1H),4.09(s,2H),3.12-2.90(m,4H), 2.44(d,J=17.1Hz,2H),2.32(d,J=17.0Hz,2H),2.18(d,J=16.2Hz,2H),1.97(d,J=16.1Hz,2H),1.27-1.13(m,6H),1.00(s,6H),0.84(s,6H). 13 C NMR(101MHz,DMSO)δ194.90(2C),153.99,149.39(2C),148.03,147.84,138.33,117.55,114 .71,111.36(2C),50.15,46.59(2C),32.05(2C),28.99(2C),26.31(4C),8.52(2C).HRMS(ESI + )m / z calcd for C 28 H 39 N2O3 + 451.2961found451.2955Da.

[0116] According to the similar synthesis method of Examples 1-10, the following compounds were prepared:

[0117]

[0118]

[0119]

[0120]

[0121]

[0122]

[0123] Example 11: In vitro testing of the protective ability of compounds against glutamate-damaged nerve cells:

[0124] This study aimed to evaluate the protective effect of compounds on glutamate-induced neuronal excitotoxicity and its dose-dependent mechanism. Primary cultured mouse cortical neurons or mouse hippocampal neuron HT22 cell lines were cultured in Neurobasal medium (containing 2% B27 and 1% GlutaMAX, neurons) or DMEM / F12 medium (HT22), respectively. The experimental groups included a blank control group (no glutamate and compound), a glutamate injury group (25mM glutamate treatment), a compound intervention group (25mM glutamate + target compound 10μM gradient concentration) and a positive control group (10mM glutamate + 10μM sefotaxime, NMDA receptor antagonist). The specific process was as follows: First, cells were seeded in 96-well plates (neuron density 5×10 4 / well, HT22 cell density 9×10 3 / well), after the neurons mature or the HT22 cell density reaches 70% confluence, 25 mM glutamate (prepared in culture medium, pH 7.4) and the target compound or sefotaxime were added simultaneously for treatment for 24 hours, respectively; after termination of the experiment, cell viability was detected by CCK-8 assay (in a dark environment, 10 μL CCK-8 reagent was added to each well, incubated at 37°C for 2 hours (neurons) / 40 minutes (HT22), the absorbance at 450 nm was measured and calculated according to the formula "survival rate (%) = [(experimental group OD - blank OD) / (control group OD - blank OD)]×100%"). In addition, cytotoxicity was detected using an LDH kit, and absorbance was measured at 490 nm. Cell damage was assessed according to the formula "Cytotoxicity or mortality (%) = (absorbance of treated sample - absorbance of sample control well) / (absorbance of maximum cell enzyme activity - absorbance of sample control well) × 100". After loading with the Fluo-4 AM fluorescent probe, the intracellular calcium ion concentration (quantitative calcium overload level) was measured by fluorescence microscopy or flow cytometry. Experimental data were expressed as mean ± SD and statistically analyzed using GraphPad Prism software. The regulatory effects of the compounds on neuronal cell survival, membrane integrity, and calcium homeostasis in the glutamate toxicity model were systematically analyzed, and their protective efficacy and potential mechanism were verified by comparison with the positive drug sefotaxime.

[0125] Table 1: Protective ability of the compounds synthesized according to the examples against glutamate-damaged nerve cells

[0126]

[0127] Example 12: In vitro testing of the protective ability of compounds on nerve cells in the OGD model:

[0128] This study aimed to evaluate the protective effect of the target compound on oxygen-glucose deprivation (OGD)-induced ischemic injury of neural cells and its dose-dependent mechanism. Primary cultured mouse hippocampal / cortical neurons or HT22 neural cell lines were cultured in Neurobasal+B27 (neurons) or DMEM high glucose medium (HT22), respectively. The experimental groups included a normal control group, an OGD model group (compound intervention group (OGD treatment + target compound 10 μM gradient concentration) and a positive control group (OGD treatment + edaravone 10 μM or safortide 10 μM). The specific process was as follows: first, the cells were seeded in a 96-well plate (neuron density 5×10 4 / well, HT22 cell density 9×10 3 Once the neurons are mature or the HT22 cell density reaches 70% confluence, remove the cells from the incubator and discard the culture medium. Add an appropriate amount of sugar-free DMEM to the culture dish or plate and disinfect the hypoxic chamber with a 75% alcohol solution. Then, move the dish or plate to a hypoxic chamber (95% N2 / 5% CO2) and incubate at 37°C for 6-8 hours. Return to complete DMEM and add the compound and positive drug treatments before incubating for an additional 24 hours. Negative control cells remain untreated and continue to incubate in complete DMEM. Finally, the cell viability was detected by CCK-8 assay (10 μL CCK-8 reagent was added to each well, incubated at 37°C in the dark for 2 hours (neurons) / 40 minutes (HT22), and the absorbance at 450 nm was measured to calculate the survival rate), the cytotoxicity was detected by LDH kit, the reactive oxygen species (ROS) level was detected by DCFH-DA fluorescent probe (10 μM loading for 30 minutes, Ex / Em=488 / 525 nm), and the early apoptosis was quantified by Annexin V-FITC / PI double staining flow cytometry (Annexin V + / PI-) and late apoptosis / necrosis (Annexin V + / PI + ) cell ratio; GraphPad Prism software was used to statistically analyze the test results (experimental data are expressed as mean ± SD). The target compounds' regulatory effects on neuronal cell survival, oxidative stress, and apoptosis in the OGD injury model were systematically analyzed, and their protective efficacy was verified by comparison with positively charged drugs. Table 2 shows the protective ability of the compounds synthesized according to the examples against neuronal cells treated with the OGD model.

[0129] Table 2: Protective ability of the compounds synthesized in the examples on nerve cells treated with OGD model

[0130]

[0131]

[0132] Example 13: In vitro surface plasmon resonance testing of compounds with ASIC1a and NMDAR proteins

[0133] This experiment used surface plasmon resonance (SPR) technology to directionally immobilize recombinant ASIC1a and NMDAR proteins on a carboxymethyl dextran chip (CM5). The proteins were fixed to the chip surface using EDC / NHS-activated amino coupling, and a reference channel was set for background subtraction. The target compound and positive control (saforin) were serially diluted (0.1-100 μM, containing 1% DMSO) and injected into the chip in a multi-cycle kinetic mode. The binding and dissociation processes were monitored (binding for 120 seconds and dissociation for 300 seconds). The chip surface was regenerated with glycine (pH 2.0). Based on the real-time response curves, two types of equilibrium dissociation constants were calculated using kinetic analysis (1:1 Langmuir model) and steady-state analysis (equilibrium binding response value-concentration fitting), respectively: ① K D (kinetic)=k off / k on , through the kinetic rate constant (k on 、k off ) characterizes dynamic binding properties; ②K D Affinity: Steady-state binding curve fitting (response value saturation platform) directly reflects overall affinity. System specificity is verified through positive controls, while DMSO concentration, protein stability, and regeneration conditions are strictly controlled to ensure data reliability. This allows for a systematic analysis of the interaction kinetics and thermodynamic characteristics between the compound and the target protein, providing two-dimensional binding parameters (dynamic rate and static affinity) for drug design. Table 3 shows the SPR test results for some selected compounds:

[0134] Table 3: SPR test results of some compounds synthesized according to the examples

[0135]

[0136]

Claims

1. A diketone compound, characterized in that: As shown in general formula (I) or general formula (II): , in the general formula (I) and formula (II): R1 is hydrogen, C1-C4 straight or branched alkyl, C1-C4 acyl, carboxyl, C1-C4 straight or branched alkylcarbonyloxy, or the following groups which are unsubstituted or substituted with 1-2 substituents: C3-C7 cycloalkyl, C5-C7 aromatic ring group, 5-7 membered aromatic heterocyclic group, 5-7 membered heterocyclic group, C2-C7 alkynyl, or C2-C7 alkenyl, wherein each of the heterocyclic group and aromatic heterocyclic group contains 1-3 heteroatoms selected from oxygen, sulfur, and nitrogen; The substituents are each independently selected from the following group: halogen, C1-C4 straight or branched alkyl, C1-C4 straight or branched alkoxy, C1-C4 straight or branched ester, cyano, nitro, hydroxy, amino, aminomethyl, dimethylamino, hydroxymethyl, trifluoromethyl, carboxyl, sulfhydryl, C2-C4 acyl, amide, aminosulfonyl, C1-C4 alkyl substituted sulfonyl, or two adjacent substituents together with the carbon atoms to which they are attached form a 5-7 membered ring; R2 is a C1-C4 straight-chain or branched alkyl group, a C1-C4 straight-chain or branched alkoxy group, a C1-C4 straight-chain or branched alkylthio group, a C1-C4 straight-chain or branched alkyl-substituted amino group, a hydroxyl group, an amino group, a C3-C7 cycloalkyl group which is unsubstituted or substituted with 1-2 substituents, or a 5-7 membered heterocyclic group; The heterocyclic groups each contain 1 to 3 heteroatoms selected from oxygen, sulfur and nitrogen; The substituents are independently selected from the following groups: halogen, C1-C4 straight chain or branched alkyl, C1-C4 straight chain or branched alkoxy, C1-C4 straight chain or branched ester, hydroxy, hydroxymethyl, trifluoromethyl, carboxyl, thiol, C2-C4 acyl, amide; R3 or R4 is hydrogen, C1-C4 straight or branched alkyl, or both together with the carbon atoms to which they are attached form a 3-7 membered ring; n is 0, 1, 2.

2. A diketone compound according to claim 1, characterized in that The compound of the general formula (I) or general formula (II) is selected from the following groups: ； ； ； ； 。 3. A diketone compound according to claim 1, characterized in that The diketone compounds represented by general formula (I) or general formula (II) or their pharmaceutically acceptable salts, solvates, hydrates, enantiomeric forms, diastereomers and racemic compounds are used to prepare (a) allosteric blockers of glutamate-sensitive acid-sensitive ion channels (ASICs), ASICs or N-methyl-D-aspartate receptor (NMDAR) signaling pathway inhibitors; and (b) drugs for treating and / or preventing or alleviating diseases related to nerve damage.

4. A diketone compound according to claim 3, characterized in that The nerve damage-related disease is selected from the following group: ischemic brain injury (such as stroke), Alzheimer's disease (AD), Parkinson's disease (PD), amyotrophic lateral sclerosis (ALS), multiple sclerosis (MS), epilepsy, spinal cord injury (SCI), glaucoma, traumatic brain injury (TBI), neuropathic pain, migraine and its complications, or a combination thereof.

5. A diketone compound according to claim 3, characterized in that The related diseases caused by nervous system disorders are selected from the following group: Alzheimer's disease, sensory or motor disorders, memory loss, myasthenia gravis, progressive muscular dystrophy, post-poliomyelitis sequelae, cerebral palsy in children, multiple neuritis and its complications, or a combination thereof.

6. The diketone compound according to claim 1, characterized in that The compound of the general formula (I) or general formula (II) or its pharmaceutically acceptable salt having a solvate, hydrate, enantiomeric form, diastereomer or racemate is one of the compounds No. 4, 11-14, 16-18, 25, 31, 34, 36, 38, 39, 41, 43 in the above group.

7. A pharmaceutical composition, characterized in that include: A therapeutically effective amount of a diketone compound represented by general formula (I) or general formula (II) or a pharmaceutically acceptable salt thereof in the form of a solvate, a hydrate, an enantiomeric form, a diastereomer or a racemate, and optionally a pharmaceutically acceptable carrier, one or more compatible solid or liquid fillers or gel substances, including but not limited to: ion exchangers, alumina, aluminum stearate, lecithin, serum proteins such as human albumin, phosphates, glycerol, sorbic acid, potassium sorbate, vegetable oils, partial glyceride mixtures of saturated vegetable fatty acids, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, calcium sulfate, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulosic materials, polyethylene glycol, sodium carboxymethyl cellulose, polyacrylates, beeswax, lanolin, gelatin, talc, solid lubricants, emulsifiers, wetting agents, colorants, flavorings, stabilizers, antioxidants, preservatives, and pyrogen-free water.

8. A pharmaceutical composition according to claim 7, characterized in that A safe amount of a diketone compound represented by general formula (I) or general formula (II), or a pharmaceutically acceptable salt, solvate, hydrate, enantiomer, diastereomer or racemate thereof, is administered to a subject in need thereof to significantly improve the condition without causing serious side effects.

9. The diketone compound according to claim 1, characterized in that The pharmaceutically acceptable salt is a conventional non-toxic salt formed by the reaction of the diketone compound represented by the general formula (I) or the general formula (II) with an inorganic acid or an organic acid, wherein the inorganic acid or the organic acid includes hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, aminosulfonic acid, phosphoric acid, citric acid, tartaric acid, lactic acid, pyruvic acid, acetic acid, benzenesulfonic acid, p-toluenesulfonic acid, methanesulfonic acid, naphthalenesulfonic acid, ethanesulfonic acid, naphthalenedisulfonic acid, maleic acid, malic acid, malonic acid, fumaric acid, succinic acid, propionic acid, oxalic acid, trifluoroacetic acid, stearic acid, pamoic acid, hydroxymaleic acid, phenylacetic acid, benzoic acid, salicylic acid, glutamic acid, ascorbic acid, p-aminobenzenesulfonic acid acid, 2-acetoxybenzoic acid and isethionic acid, oleic acid, linoleic acid, linolenic acid, etc.; or sodium salt, potassium salt, calcium salt, aluminum salt or ammonium salt formed by the compound of general formula (I) or general formula (II) with an inorganic base; or methylamine salt, ethylamine salt or ethanolamine salt formed by the compound of general formula (I) or general formula (II) with an organic base; or the compound of general formula (I) or general formula (II) forms an ester with lysine, arginine or ornithine and then forms the corresponding inorganic acid salt with hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid or phosphoric acid, or the corresponding organic acid salt formed with formic acid, acetic acid, picric acid, methanesulfonic acid or ethanesulfonic acid.

10. A pharmaceutical composition according to claim 7, characterized in that A safe dose is administered to a subject in need thereof. For a person weighing 60 kg, the daily dose is usually 1 to 2000 mg.

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