A compound for treating epilepsy and a preparation method thereof

Abstract

The application provides a compound for treating epilepsy and a preparation method thereof. The compound of the application exhibits significant therapeutic effect in the field of epilepsy treatment, has a strong and effective inhibitory effect on the onset of epilepsy, and has extremely excellent therapeutic effect and leads the CBD group in all indexes according to the data analysis of the detection of the expression of apoptosis-related proteins from the evaluation of the molecular biology method. In addition, the compound can effectively prolong the latency of the onset of epilepsy, thereby reducing the frequency of taking medicine and greatly reducing the toxic side effects possibly caused by the medicine. The new compound provided by the application improves the safety and effectiveness of the treatment of epilepsy in many aspects, and opens up a new direction with great potential in the field of epilepsy treatment.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, specifically to a compound for treating epilepsy and its preparation method. Background Technology

[0002] Epilepsy is one of the most common chronic neurological disorders. During a seizure, abnormal neuronal discharges occur, and patients may experience symptoms such as confusion, convulsions, and mental abnormalities. It is a disease that seriously threatens global human health and safety, with approximately five million people diagnosed worldwide each year.

[0003] Elucidating the pathogenesis of epilepsy has been a research hotspot in recent years. Currently, it is generally believed that epilepsy occurs due to an increase in the levels of excitatory neurotransmitters or a decrease in the levels of inhibitory neurotransmitters. However, it remains unclear which neurotransmitter plays a major role in the development of epilepsy. As one aspect of the epilepsy pathogenesis, the regulation of γ-aminobutyric acid (GABA) levels and abnormal expression or impaired function of GABA receptors are closely related to epileptic seizures.

[0004] In recent years, drugs targeting different pathogenesis mechanisms of epilepsy have been developed, and more than 20 anti-epileptic drugs are now used in clinical practice. However, about 20-30% of patients still cannot achieve effective control of epileptic seizures with drugs and may develop drug resistance. Therefore, there are still huge challenges in the drug treatment of epilepsy patients. Summary of the Invention

[0005] To overcome the shortcomings of the prior art, the present invention provides a compound for treating epilepsy and a method for preparing the same.

[0006] In a first aspect of the present invention, a compound is provided having the following structure:

[0007]

[0008] in,

[0009] R1 is selected from C l-10 alkyl;

[0010] Furthermore, R1 is selected from: -CH3, -CH2CH3, -CH2CH2CH3, -(CH2)3CH3, -(CH2)4CH3, -CH(CH2)CH2CH3, -CH(CH2)CH(CH2)CH3;

[0011] Preferably, R1 is selected from: -CH2CH2CH3, -(CH2)3CH3, -(CH2)4CH3;

[0012] More preferably, R1 is -(CH2)4CH3.

[0013] X1 and X2 are independently selected from single-bonded, alkylene (e.g., C1) bonds. 1-10 Alkylene, C 1-5 Alkylenes), alkylenes separated by one or more -C(=O)O-, alkylenes separated by one or more -OC(=O)-, alkylenes separated by one or more -OC(=O)O-, and alkylenes separated by a combination of the above spacer groups;

[0014] Furthermore, X1 and X2 are independently selected from: -CH2-, -CH2CH2-, -(CH2)2CH2-, -(CH2)3CH2-, -CH2CH(CH3)2-, -CH(CH3)CH2CH2-, -CH2C(=O)OCH2-, -CH2CH2OC(=O)CH2CH2-, -CH2CH2C(=O)OCH2CH2-, -CH2CH2OC(=O)CH2CH(CH3)2-, -CH2CH(CH3)2OC(=O)OCH2-, -CH2CH2C(=O)OCH2CH2OC(=O)CH2-;

[0015] Furthermore, X1 is C 1-5 Alkyl groups, such as -CH2-, -CH2CH2-, -(CH2)2CH2-, -(CH2)3CH2-; further, X2 is a C separated by -C(=O)O-. 1-5 Alkylene or C separated by -OC (=O)- 1-5 Alkyl groups, such as -CH2OC(=O)CH2-, -CH2C(=O)OCH2-, -CH2CH2OC(=O)CH2CH2-, -CH2CH2C(=O)OCH2CH2-;

[0016] Preferably, X1 is -CH2CH2- and X2 is -CH2CH2OC(=O)CH2CH2-.

[0017] R2 and R3 are independently selected from: -H, -OH, -COOH, X3 is selected from CH2, NH, O, and S;

[0018] Preferably, R2 and R3 are independently selected from: -H,

[0019] More preferably, R2 and R3 are independently selected from: -H, More preferably, both R2 and R3 are In one embodiment of the present invention, the compound has the following structure:

[0020]

[0021] In a second aspect of the invention, a stereoisomer of the compound described in the first aspect is provided, having the following structure:

[0022]

[0023] In one embodiment of the present invention, the structure of the stereoisomer is as follows:

[0024]

[0025] In a third aspect of the invention, a salt or crystal form of the compound described in the first aspect or the stereoisomer described in the second aspect is provided, particularly a pharmaceutically acceptable salt.

[0026] Furthermore, the salt is either a base addition salt or an acid addition salt.

[0027] The alkali addition salts can be formed by metals or amines with compounds, especially alkali metal salts and alkaline earth metal salts, such as sodium salts, potassium salts, magnesium salts, and calcium salts;

[0028] The acid addition salt can be formed by inorganic or organic acids and compounds, such as hydrochloride, nitrate, sulfate, phosphate, citrate, formate, fumarate, maleate, acetate, oxalate, succinate, tartrate, methanesulfonate, p-toluenesulfonate, preferably oxalate.

[0029] In a fourth aspect of the invention, a method for preparing a compound is provided, comprising reacting compound I with 1-1 to obtain compound II;

[0030] Compound II undergoes transesterification to yield compound IV;

[0031] Alternatively, compound II can be reacted with a diol, followed by transesterification, to yield compound IV. The specific synthetic route is as follows:

[0032]

[0033] Where R5 is C 1-6 alkyl;

[0034] X4 is selected from alkylene groups (e.g., C). 1-10 Alkylene, C 1-5 (alkylene).

[0035] In some embodiments of the present invention, the preparation method includes the following steps:

[0036] 1) Dissolve compound I, add 1-1, a desiccant and a catalyst, and react to obtain compound II;

[0037] 2) Dissolve compound II, add an alkaline reagent and a diol, react to obtain compound III;

[0038] 3) Dissolve Ⅲ, add 1-2, alkaline reagent, condensing agent and catalyst, react to obtain the compound described in the first aspect (compound Ⅳ);

[0039] The terms 1-2 are:

[0040] R4 is selected from: -H, -OH, -COOH, X3 is selected from CH2, NH, O, and S.

[0041] Specifically, the reaction in step 1) is carried out under the protection of an inert gas.

[0042] Preferably, the inert gas is selected from nitrogen or argon.

[0043] Specifically, the solvent used for dissolving in step 1) is at least one of tetrahydrofuran, dioxane, toluene, xylene, dichloromethane, dichloroethane, ethyl acetate, or isopropyl acetate.

[0044] Preferably, the desiccant is one of anhydrous magnesium sulfate, anhydrous sodium sulfate, anhydrous calcium sulfate, anhydrous copper sulfate, or calcium chloride.

[0045] Preferably, the catalyst in step 1) is one of boron trifluoride diethyl ether, aluminum trichloride, aluminum tribromide, ferric chloride, antimony pentachloride, hydrofluoric acid, or phosphorus pentoxide, and more preferably boron trifluoride diethyl ether.

[0046] Preferably, the molar ratio of I to 1-1 is 1:0.5 to 1:1.5, for example, 1:0.5, 1:0.7, 1:0.9, 1:1.0, 1:1.1, 1:1.2, 1:1.3, 1:1.4 or 1:1.5, etc.

[0047] Preferably, the molar ratio of I to the catalyst is 1:0.01 to 1:0.15, for example, 1:0.01, 1:0.03, 1:0.05, 1:0.07, 1:0.08, 1:0.09, 1:0.10, 1:0.11, 1:0.13 or 1:0.15.

[0048] Preferably, the reaction temperature in step 1) is -40 to -10°C, for example -40°C, -35°C, -30°C, -25°C, -20°C, -15°C, or -10°C.

[0049] Preferably, the reaction time in step 1) is 0.5 to 6 hours, for example, 0.5 hours, 1 hour, 1.5 hours, 2 hours, 2.5 hours, 3 hours, 4 hours, 5 hours or 6 hours.

[0050] Specifically, after the reaction in step 1) is completed, a quenching agent is added for quenching.

[0051] Preferably, the quenching agent is an organic amine, a Lewis base, a polar solvent, or a weak base solution, such as ethylenediamine, triethylamine, ammonia, ethanol, water, a saturated sodium bicarbonate solution, or a saturated potassium bicarbonate solution.

[0052] Specifically, step 1) further includes a purification step, which includes filtration, extraction, drying, concentration, and column chromatography.

[0053] Preferably, the solvent used for extraction is one or more of dichloromethane, trichloromethane, ethyl acetate, tetrahydrofuran, or isopropanol.

[0054] In one embodiment of the present invention, the elution conditions for column chromatography in step 1) are hexane: ethyl acetate = 100:0.5 to 100:2.

[0055] Specifically, the solvent used for dissolving III in step 2) is at least one of methanol, ethanol, isopropanol, tetrahydrofuran, dioxane, dimethyl sulfoxide, dimethylformamide, dimethylacetamide, toluene, or acetonitrile.

[0056] Preferably, the alkaline reagent is at least one selected from triethylamine, butylamine, pentylamine, hexylamine, diethylamine, dibutylamine, dipentylamine, tributylamine, aniline, N-methylaniline, N,N-diisopropylethylamine, imidazole, pyrrole, piperazine, pyrazine, pyridine, sodium methoxide, sodium ethoxide, sodium bicarbonate, potassium bicarbonate, ammonium bicarbonate, sodium carbonate, potassium carbonate, cesium carbonate, lithium carbonate, cesium carbonate, ammonium carbonate, sodium hydroxide, lithium hydroxide, potassium hydroxide, cesium hydroxide, or barium hydroxide.

[0057] Preferably, the diol is one of ethylene glycol, propylene glycol, butanediol, or pentanediol.

[0058] Specifically, the reaction in step 2) is carried out under negative pressure.

[0059] Preferably, the pressure of the negative pressure environment is -0.9 to -0.1 bar, such as -0.9 bar, -0.8 bar, -0.7 bar, -0.6 bar, -0.5 bar, -0.4 bar, -0.3 bar, -0.2 bar, or -0.1 bar.

[0060] Preferably, the reaction temperature in step 2) is 90 to 140°C, such as 90°C, 100°C, 110°C, 120°C, 125°C, 130°C, 135°C or 140°C.

[0061] Preferably, the reaction time in step 2) is 1 to 6 hours, for example, 1 hour, 1.5 hours, 2 hours, 2.5 hours, 3 hours, 4 hours, 5 hours or 6 hours.

[0062] Specifically, step 2) further includes a purification step, which includes extraction and column chromatography.

[0063] In one embodiment of the present invention, the purification method in step 2) is column chromatography, and the elution conditions of column chromatography are hexane: ethyl acetate = 10:1 to 4:1.

[0064] Preferably, the solvent required to dissolve III is at least one selected from dichloromethane, tetrahydrofuran, N,N-dimethylformamide, 2-methyltetrahydrofuran, dimethyl sulfoxide, tert-butyl methyl ether, ethyl acetate, isopropanol, or dimethyl carbonate.

[0065] Preferably, the catalyst in step 3) is at least one of N-hydroxysuccinimide (HOSu), N-hydroxy-7-azabenzotriazole (HOAt), 1-hydroxybenzotriazole (HOBt) or 4-dimethylaminopyridine (DMAP).

[0066] Preferably, the condensing agent is at least one selected from N,N'-dicyclohexylcarbodiimide (DCC), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI), N,N'-carbonyldiimidazole (CDI), N,N'-diisopropylcarbodiimide (DIC), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (HATU), O-benzotriazole-tetramethylurea hexafluorophosphate (HBTU), propyl tricyclic phosphate anhydride (T3P), n-butyl phosphate anhydride (T4P), diphenyl azidophosphate (DPPA), or bis(2-oxo-3-oxazolyl)hypophosphine chloride (BOP-Cl).

[0067] In one embodiment of the present invention, the catalyst is selected from 4-dimethylaminopyridine (DMAP), and the condensing agent is selected from N,N'-dicyclohexylcarbodiimide (DCC).

[0068] Specifically, the reaction temperature in step 3) is 10 to 35°C, for example, 10°C, 15°C, 20°C, 25°C, 30°C, or 35°C.

[0069] Specifically, the reaction time in step 3) is 8 to 24 hours, for example, 8 hours, 12 hours, 14 hours, 16 hours, 18 hours, 20 hours, 22 hours, or 24 hours.

[0070] Preferably, the molar ratio of Ⅲ to 1-2 is 1:1.0 to 1:5.0, for example, 1:1.0, 1:1.5, 1:2.0, 1:2.5, 1:3.0, 1:3.5, 1:4.0, 1:4.5 or 1:5.0, and more preferably 1:2.5 to 1:4.5.

[0071] Preferably, the molar ratio of III to the alkaline reagent is 1:2.0 to 1:8.0, for example, 1:2.0, 1:3.0, 1:3.5, 1:4.0, 1:4.5, 1:5.0, 1:6.0, 1:7.0 or 1:8.0, etc.

[0072] Preferably, the molar ratio of III to the condensing agent is 1:1.5 to 1:4.5, for example, 1:2.0, 1:2.5, 1:3.0, 1:3.5, 1:4.0 or 1:4.5.

[0073] Preferably, the molar ratio of III to the catalyst is 1:0.01 to 1:0.15, such as 1:0.01, 1:0.03, 1:0.05, 1:0.07, 1:0.09, 1:0.1, 1:0.12, 1:0.14 or 1:0.15.

[0074] Specifically, step 3) further includes a purification step, which includes filtration, concentration, and column chromatography.

[0075] In one embodiment of the present invention, the elution conditions for column chromatography in step 3) are hexane: ethyl acetate = 10:1 to 2:1.

[0076] Specifically, the preparation method also includes a salt-forming method.

[0077] Specifically, the salt formation method includes: dissolving compound IV, slowly adding an acidic or alkaline solution at low temperature, filtering and drying to obtain the corresponding salt form of compound IV.

[0078] Preferably, the salt form of compound IV is a base addition salt or an acid addition salt.

[0079] In one embodiment of the present invention, the salt form of compound IV is an oxalate.

[0080] Preferably, the drying method is one or more of vacuum drying, freeze drying, and natural drying.

[0081] In one embodiment of the present invention, the method for preparing the compound is as follows:

[0082] 1) Dissolve raw material I, add raw material 1-1, desiccant and catalyst, and react at low temperature under inert gas protection; after the reaction is completed, quench, filter, extract, dry, concentrate, and column chromatography to finally obtain compound II;

[0083] 2) Dissolve compound II, add an alkaline reagent and a diol, and react at high temperature under low pressure. After the reaction is complete, extract and perform column chromatography to obtain compound III.

[0084] 3) Dissolve Ⅲ, add raw materials 1-2, alkaline reagent, condensing agent and catalyst, react at room temperature, filter, concentrate and column chromatography to finally obtain compound Ⅳ;

[0085] 4) Dissolve N, slowly add acid or alkaline solution at low temperature, filter and dry to obtain the corresponding compound N salt form.

[0086] A fifth aspect of the present invention provides the use of a compound or a pharmaceutically acceptable salt or stereoisomer thereof in the preparation of a medicament for treating epilepsy.

[0087] In a sixth aspect of the invention, a pharmaceutical composition is provided comprising the compound described herein or a pharmaceutically acceptable salt or stereoisomer thereof, and one or more pharmaceutically acceptable excipients.

[0088] Specifically, the pharmaceutically acceptable excipients may be selected from one or more combinations of carriers, excipients, diluents, lubricants, wetting agents, emulsifiers, preservatives, antioxidants, buffers, antibacterial agents, suspending agents, suspending aids, solubilizers, thickeners, stabilizers, sweeteners, and flavorings.

[0089] Specifically, the compound or its pharmaceutically acceptable salts or stereoisomers may be used alone or in combination with other types of active ingredients.

[0090] Preferably, the pharmaceutical composition may contain the compound in a weight ratio of 0.01 to 99.5% (specifically, 0.01%, 0.1%, 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 99%, 99.5%).

[0091] Specifically, the above-mentioned pharmaceutical compositions can be in any dosage form or administration method, including but not limited to tablets (including sugar-coated tablets, film-coated tablets, sublingual tablets, orally disintegrating tablets, oral tablets, etc.), pills (water pills, honey pills, paste pills, concentrated pills), powders, granules, capsules (including soft capsules, microcapsules), lozenges, syrups, solutions, emulsions, suspensions, controlled-release formulations (e.g., instantaneous-release formulations, sustained-release formulations, sustained-release microcapsules), aerosols, and films (e.g., oral). Disintegrating film preparations, oral mucosa-adhesive film preparations, injections (e.g., subcutaneous injection, intravenous injection, intramuscular injection, intraperitoneal injection), intravenous infusions, transdermal absorption preparations, ointments, lotions, adhesive preparations, suppositories (e.g., rectal suppositories, vaginal suppositories), nasal preparations, pulmonary preparations (inhalation), eye drops, etc.; forms of administration, such as oral administration or parenteral administration (e.g., intravenous, intramuscular, subcutaneous, intra-organ, intranasal, intradermal, infusion, intracerebral, rectal, etc.).

[0092] A seventh aspect of the present invention provides a method for treating epilepsy, the method comprising administering to a subject in need an effective dose of the compound of the present invention or a pharmaceutically acceptable salt, stereoisomer, or pharmaceutical composition thereof of the present invention.

[0093] Specifically, the effective dose depends on many factors, including the patient's age, weight, gender, natural health condition, nutritional status, drug activity intensity, administration time, metabolic rate, severity of illness, etc.

[0094] The term "comprising" as used in this invention is an open-ended description, encompassing the specified ingredients or steps described, as well as other specified ingredients or steps that do not substantially affect the description.

[0095] The term "treatment" as used in this invention refers to slowing down, interrupting, preventing, controlling, stopping, alleviating, reducing, or reversing a sign, symptom, disorder, condition, or progression or severity of a disease after it has begun to develop, but does not necessarily involve the complete elimination of all disease-related signs, symptoms, conditions, or disorders.

[0096] The “effective amount” as used in this invention refers to the amount or dose of the product of this invention that provides the desired treatment or prevention after being administered to a patient or organ in one or more doses.

[0097] The "subject" described in this invention can be a human or a non-human animal, such as a non-human mammal. The non-human mammal can be a wild animal, a zoo animal, an economically important animal, a pet, or a laboratory animal, etc. Preferably, the non-human mammal includes, but is not limited to, pigs, cattle, sheep, horses, donkeys, foxes, minks, jackals, camels, dogs, cats, rabbits, mice (e.g., rats, mice, guinea pigs, hamsters, gerbils, chinchillas, squirrels), or monkeys, etc.

[0098] The beneficial effects of this invention are:

[0099] This invention provides a novel compound that demonstrates significant efficacy in the treatment of epilepsy, exhibiting a powerful and effective inhibitory effect on epileptic seizures. Evaluation using the Ono's grading system successfully induced rats experiencing clonic seizures and hindlimb standing to transition to a seizure-free state within 7 days, strongly demonstrating the compound's excellent efficacy in controlling the progression of epileptic seizures. Molecular biological evaluation and analysis of apoptosis-related protein expression data show that this compound exhibits extremely superior therapeutic effects, with all indicators surpassing those of the CBD group.

[0100] Furthermore, this compound can effectively prolong the latency period of epileptic seizures, thereby reducing the frequency of medication and significantly lowering the potential toxic side effects. Simultaneously, by modifying its structure, compound 1 exhibits approximately 40% improved solubility compared to cannabidiol. This improvement not only effectively enhances drug solubility but also increases bioavailability and strengthens the compound's stability, creating favorable conditions for drug formulation development.

[0101] The novel compounds provided by this invention improve the safety and effectiveness of epilepsy treatment in many ways, opening up a new and highly promising direction for the field of epilepsy treatment. Attached Figure Description

[0102] Figure 1 Compound 1 oxalate 1 H-NMR spectrum. Detailed Implementation

[0103] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0104] Reagents used in the examples: Pentylenetetrazole (PTZ): Siama; Product No.: P6500-25G; Gamma-aminobutyric acid (GABA): Shanghai Tongwei Industrial Co., Ltd.; Catalog No.: TW1442; Rat Gly Elisa Kit: Wuhan Bailehui Biotechnology Co., Ltd.; Catalog No.: orb782652; Colorimetric L-Aspartate (Aspartic Acid) Assay Kit: Wuhan AmyJet Technology Co., Ltd.; Catalog No.: AAT Bioquest.

[0105] Preparation of reagents in the examples:

[0106] a. PTZ solution: Weigh 525.2 mg of PTZ, dissolve it in physiological saline, and prepare a drug solution with a concentration of 17.5 mg / ml.

[0107] b. Sodium valproate solution: Weigh 1640.4 mg of sodium valproate, dissolve it in purified water, and prepare a drug solution with a concentration of 20 mg / ml.

[0108] c. Solutions of each compound: Compound 1 is the oxalate form of compound 1, weigh 1640.3 mg, and compound 2 is cannabidiol (CBD), weigh 1640.4 mg. They are dissolved in MCT oil to prepare a drug solution with a concentration of 20 mg / ml.

[0109] Animals used in the examples: SD rats, 5-6 weeks old, male, SPF grade, purchased from Spiford (Beijing) Biotechnology Co., Ltd., experimental animal batch number: 110324220103746757.

[0110] The rearing conditions in this example were as follows: Three animals of the same sex were housed in each cage in an SPF-grade animal room with a room temperature controlled between 20°C and 26°C and a relative humidity controlled between 40% and 70%. Light was provided in alternating light and dark conditions for 12 hours per day. The bedding was high-pressure sterilized corn cob-based experimental material. Except when fasting was required, animals were fed qualified rodent feed daily with free access to food. During quarantine, training, and the experiment, purified water was provided via drinking bottles with free access to water. The drinking bottles and stoppers were autoclaved before use.

[0111] Example 1: Preparation of Compound 1

[0112]

[0113] first step

[0114] (1'R,2'R)-2,6-dihydroxy-5'-methyl-4-pentyl-2'-(prop-1-en-2-yl)-1',2',3',4'-tetrahydro-[1,1'-biphenyl]-3-carboxylic acid

[0115] Ethyl acetate

[0116] Add (1S,4R)-1-methyl-4-(1-methylvinyl)-2-cyclohexen-1-ol (compound i, 6.62 g, 43.56 mmol), ethyl 2,4-dihydroxy-6-pentylbenzoate (10 g, 39.6 mmol), and 400 mL of dry dichloromethane to a 1 L single-necked flask. Stir at room temperature until the reactants are completely dissolved, then add anhydrous magnesium sulfate (14.4 g). Place the reaction flask in a dry ice-ethanol bath under nitrogen protection, controlling the external temperature at around -20 °C. After the reaction temperature stabilizes, add boron trifluoride diethyl ether (1.06 mL, 3.96 mmol) to the reaction flask and maintain the reaction temperature for 2 h. After extracting and separating the layers with a small amount of the reaction solution using saturated sodium bicarbonate solution, the dichloromethane layer was sampled and monitored by TLC. Once the reaction was confirmed to be complete, the solids in the reaction solution were filtered off. The filtrate was transferred to a 1L separatory funnel, and approximately 500ml of saturated sodium bicarbonate solution was added. After thorough mixing, the mixture was allowed to stand in the separatory funnel until separation was complete. The aqueous layer was separated, and the dichloromethane layer was washed once more with saturated sodium chloride solution. After extraction, the aqueous layer was discarded. The dichloromethane layer was dried with anhydrous sodium sulfate for 30 minutes. After drying, the anhydrous sodium sulfate was filtered off. The filtrate was concentrated under reduced pressure by rotary evaporation until it no longer decreased in weight, yielding approximately 18g of a pale yellow oily crude product. Silica gel column chromatography (n-hexane:ethyl acetate = 100:0.5-100:2) yielded compound ii, a colorless oily product (15.35g, 100%), with a purity of 90%.

[0117] Compound ii was confirmed by LCMS and H-NMR, and the results are shown below.

[0118] MS (ESI, positive): 387 [M+H] + ;

[0119] 1 H-NMR (CDCl3, 400MHz): δ12.10 (1H, s), 6.49 (1H, s), 6.20 (1H, s), 5.55 (1H, s) ,4.40(1H,d),4.38-4.34(3H,m),4.10-4.08(1H,m),2.88-2.83(1H,m),2.78- 2.73 (1H, m), 2.39-2.37 (1H, m), 2.22 (1H, m), 2.11-2.06 (1H, m), 1.82 (3H, s), 1.78 (3H, s), 1.74-1.63 (4H, m), 1.56 (3H, t), 1.53-1.42 (4H, m), 0.88 (3H, t).

[0120] Step 2

[0121] 2-Hydroxyethyl(1'R,2'R)-2,6-dihydroxy-5'-methyl-4-pentyl-2'-(prop-1-en-2-yl)-1',2',3',4'-tetrahydro-[1,1'-biphenyl]-3-carboxylic acid ester

[0122] Compound ii (8.52 g, 22 mmol), ethylene glycol (40 ml), and potassium hydroxide (1.33 g, 23.84 mmol) were added to a 100 ml single-necked flask. The flask was placed in an oil bath and stirred, with the external temperature controlled at approximately 120 °C. A vacuum pressure of 0.5 bar was maintained using a water pump. After the reaction temperature stabilized, the reaction was allowed to proceed for 2 hours. A small amount of the reaction solution was taken for monitoring and TLC to confirm the completion of the reaction. The flask was then allowed to cool to room temperature and diluted with 200 ml of dichloromethane. The diluted reaction solution was transferred to a 500 ml separatory funnel and washed with saturated sodium chloride solution (200 ml × 3). After extraction, the aqueous layer was discarded. The dichloromethane layer was dried with anhydrous sodium sulfate for 30 minutes. After drying, the anhydrous sodium sulfate was filtered off. The filtrate was concentrated under reduced pressure by rotary evaporation until it no longer decreased in weight, yielding approximately 9.3 g of a reddish-brown oily crude product. Silica gel column chromatography (n-hexane:ethyl acetate = 10:1-4:1) yielded compound iii, a red oily product (6 g, 67.6%), purity: 95%.

[0123] Compound iii was confirmed by LCMS and H-NMR, and the results are shown below.

[0124] MS (ESI, positive): 403 [M+H] + ;

[0125] 1 H-NMR (CDCl3, 400MHz): δ11.88 (1H, s), 6.53 (1H, s), 6.22 (1H, s), 5.52 (1H, s), 4.52 (1 H, s), 4..47-4.45 (2H, m), 4.38 (1H, s), 4.11-4.08 (1H, m), 3.97-3.94 (2H, t), 2.89-2. 81(1H,m),2.97-2.73(1H,m),2.39-2.37(1H,m),2.21(1H,m),2.07-2.04(1H,m),1.82 -1.71 (2H, m), 1.81 (3H, s), 1.76 (3H, s), 1.54-1.52 (2H, m), 1.33 (5H, m), 0.88 (3H, t).

[0126] Step 3

[0127] 2-((3-morpholinopropionyl)oxy)ethyl(1'R,2'R)-2-hydroxy-5'-methyl-6-(3-morpholinopropionyl)oxy)-4-pentyl-2'-(prop-1-en-2-yl)-1',2',3',4'-tetrahydro-[1,1'-biphenyl]-3-carboxylic acid ester

[0128] A stir bar was placed in a 250 mL single-necked flask, and compound iii (2.3 g, 5.72 mmol) and 100 mL of dichloromethane were added. A magnetic stirrer was turned on to dissolve all the reactants. At room temperature, 3-morpholine propionate (3.37 g, 17.16 mmol), triethylamine (3.2 mL, 22.88 mmol), dicyclohexylcarbodiimide (4.1 g, 20 mmol), and 4-dimethylaminopyridine (70 mg, 0.57 mmol) were added to the reaction flask. After the addition was complete, the reaction was stirred overnight at room temperature. The insoluble matter generated during the reaction was filtered off. The filtrate was concentrated under reduced pressure by rotary evaporation to obtain an oily crude product. 100 mL of n-hexane was added to this product, and the mixture was stirred thoroughly for 30 minutes to disperse the insoluble matter evenly. The insoluble matter was filtered off again, and the filtrate was concentrated under reduced pressure by rotary evaporation to obtain 6 g of a pale yellow oily crude product. Purification was performed by silica gel column chromatography. The column was packed with 200-300 mesh silica gel using a wet packing method, and the sample was loaded using a wet packing method. The elution conditions were hexane:ethyl acetate = 10:1 to 2:1. After collecting the purity point, the chromatographic solution was concentrated to obtain 850 mg of compound 1, a colorless oily product with an HPLC purity of 97.0%.

[0129] 850 mg of compound 1 was added to a 100 mL single-necked flask, followed by 5 mL of acetone. The mixture was stirred until completely dissolved, and the flask was cooled in an ice-water bath. 8 mL of acetone was measured, and 313 mg of solid oxalic acid was added. The mixture was stirred until completely dissolved to prepare an oxalic acid solution. This oxalic acid solution was slowly added dropwise to the flask cooled in the ice-water bath using a constant-pressure dropping funnel. The addition process was maintained in an ice-water bath. After the addition was complete, the pH of the solution in the flask was measured to be 3–4. The mixture was then stirred in the ice-water bath for 1 hour. The resulting white solid was gel-like and could not be stirred or filtered. The solvent in the flask was evaporated to dryness under reduced pressure, dissolved in a small amount of methanol, and then concentrated again to dryness using a rotary evaporator. After high-vacuum drying with an oil pump, a pale yellow powder was obtained. The filter cake was thoroughly dried to obtain a white solid product (780 mg, 19.9%) with an HPLC purity of 95.7% (containing 1.83% oxalic acid).

[0130] Compound 1, oxalate type, was analyzed by LCMS and 1 H-NMR confirmed, 1 H-NMR spectrum as shown Figure 1 As shown.

[0131] MS (ESI, positive): 685 [M+H] + ;

[0132] After salt formation: 1 H-NMR (CD3OD, 400MHz): δ6.52 (1H, s), 5.12 (1H, s), 4.63 (2H, m), 4.51-4.42 (4H, m), 4.08(1H,m), 3.95-3.90-3.88(8H,m), 3.51-3.46(6H,m), 3.36(4H,m), 3.25-3.19(4 H, m), 2.95-2.79 (5H, m), 2.68 (1H, m), 2.22 (1H, m), 2.07-2.04 (1H, m), 1.78 (2H, m), 1.68(6H,m), 1.57(2H,m), 1.58-1.55(2H,t), 1.37-1.34(4H,m), 0.93-0.90(3H,t).

[0133] Example 2: Pharmacological activity study of the compound against PTZ-induced chronic epilepsy in SD rats

[0134] 1. Model Construction

[0135] 1.1 Test Methods

[0136] Prepare a fresh 1.75% PTZ solution using physiological saline. Inject the prepared PTZ solution intraperitoneally at a dose of 35 mg / kg. Administer PTZ once every other day. Observe behavioral changes in rats within 1 hour after PTZ injection, and record the latency period, seizure grade, and seizure duration.

[0137] 1.2 Test Results

[0138] Behavioral seizure indicators were graded according to Ono's classification, as shown in Table 1. Rats with five consecutive records of grade 2 or above were considered to have a successful model.

[0139] Table 1: Ono's classification of PTZ ignition attacks

[0140] Ono's rating Seizure manifestations 0 No seizures 1 Nodding or head twitching 2 Generalized myoclonus 3 Head twitching and forelimb clonus 4 Clonic seizures with hindlimb standing 5 fall 6 Generalized tonic-clonic seizures

[0141] 2. Animal grouping and dosage

[0142] After successful modeling, the experimental animals were randomly divided into groups of 8. Using appropriate disposable sterile syringes and matching gavage needles, the drugs were administered by gavage once daily for 2 weeks. The model group received physiological saline, the positive control group received sodium valproate by gavage (100 mg / kg), the compound 1 group received compound 1 oxalate prepared in Example 1 by gavage (100 mg / kg, based on the pure compound 1), and the compound 2 group received CBD by gavage (100 mg / kg). The administration continued for 2 weeks after grouping. Grouping and administration details are shown in Table 2.

[0143] Table 2. Animal grouping and drug dosage

[0144]

[0145] Before administration, and on days 7 and 14 after administration, the prepared PTZ solution was injected intraperitoneally 1 hour after gavage administration, with an injection dose of 35 mg / kg.

[0146] Clinical observation: During the trial, observations were conducted at least twice a day (once in the morning and once in the afternoon). The observations included, but were not limited to, death, onset of disease, respiration, secretions, feces, and diet and water intake.

[0147] Detection indicators: Record Ono's grade before and after drug administration (according to Table 1), record the latency period, grade and duration of the attack;

[0148] The levels of γ-aminobutyric acid, glutamate, and aspartate in the brain tissue of rats after drug administration were detected.

[0149] The expression of Bax and Bcl-2 proteins was detected by Western blotting (WB) in rat hippocampus, and the expression levels of each group were compared.

[0150] 3. Experimental Results

[0151] 3.1 Ono's rating

[0152] Before administration, on day 7 and day 13 after administration, rats in each group were intraperitoneally injected with PTZ solution (35 mg / kg) 1 hour after gavage administration. The Ono's grading before and after administration was recorded, as well as the latency period, grading, and duration of the attack.

[0153] The experimental results showed that after 7 days of treatment, compared with the model control group, the Ono's score of all experimental groups showed a decreasing trend, with compound 2 showing a significant decrease (3.4 vs. 0.7; P<0.05). After 13 days of treatment, compared with the model control group, the Ono's score of both compound 1 and compound 2 groups showed a decreasing trend, with compound 1 (2.6 vs. 0.3; P<0.05) and compound 2 (2.6 vs. 0.7; P<0.05) showing significant decreases, and the differences were statistically significant. See Tables 3 and 4 for details.

[0154] Table 3. Ono's classification (7 days of treatment)

[0155]

[0156] Note: P-value: Model group vs. results of each group after 7 days of treatment.

[0157] Table 4. Ono's classification (after 13 days of treatment)

[0158]

[0159] Note: P-value: Model group vs. results of each group after 13 days of treatment.

[0160] 3.2 Incubation period

[0161] The experiment found that after 7 days of treatment, compared with the model control group, the latency time of epileptic seizures in all experimental groups showed a prolonged trend, with compound 2 group (48.1 vs. 21.8) showing a significantly prolonged latency time. After 13 days of treatment, compared with the model control group, the latency time of epileptic seizures in all experimental groups showed a prolonged trend, with compound 1 group (55.0 vs. 25.3) and compound 2 group (50.3 vs. 25.3) showing significantly prolonged latency times, and the differences were statistically significant. See Table 5 for details.

[0162] Table 5. Seizure latency (min)

[0163]

[0164] Note: P-value: model control group vs. each treatment group

[0165] 3.3 Molecular biological detection results

[0166] 1) γ-aminobutyric acid content

[0167] The experimental results showed that after 13 days of treatment, compared with the model control group, the content of γ-aminobutyric acid in each experimental group showed an increasing trend. Among them, the positive control group (22.08 vs. 29.31; P<0.05), compound 1 group (22.08 vs. 29.96; P<0.05), and compound 2 group (22.08 vs. 29.15; P<0.01) showed significant increases, and the differences were statistically significant (see Table 6).

[0168] Table 6. Results of γ-aminobutyric acid content detection

[0169]

[0170] 2) Glutamic acid content

[0171] After 13 days of treatment, compared with the model control group, the levels of glutamate and aspartate in all experimental groups showed a decreasing trend. Among them, the levels of glutamate were significantly lower in the positive control group (139.9 vs. 108.5; P<0.05), compound 1 group (139.9 vs. 105.9; P<0.05), and compound 2 group (139.9 vs. 106.2; P<0.05), and the differences were statistically significant. The results are shown in Table 7.

[0172] Table 7. Glutamic Acid Content Detection Results

[0173]

[0174] 3) Aspartic acid content

[0175] Regarding the aspartic acid content, the positive control group (420.9 vs. 323.3; P<0.05) and compound 1 group (420.9 vs. 328.6; P<0.05) showed a significant decrease, and the difference was statistically significant. The results are shown in Table 8.

[0176] Table 8. Results of Aspartic Acid Content Detection

[0177]

[0178]

[0179] 4) Bax and Bcl-2 protein content and ratio

[0180] Western blot analysis of Bax and Bcl-2 protein expression in each experimental group revealed that, compared with the model control group, the expression of Bax and Bcl-2 proteins in each experimental group was consistent, with all groups showing an increased Bcl-2 / Bax ratio. The ratios in the positive control group (0.18 vs. 0.24; P<0.05), compound 1 group (0.18 vs. 0.41; P<0.05), and compound 2 group (0.18 vs. 0.24; P<0.05) were significantly higher, and the differences were statistically significant (see Table 9 for details).

[0181] Table 9. Results of Bax and Bcl-2 protein expression analysis in each experimental group

[0182]

[0183] The therapeutic effects of the drugs on seizures in a rat model of epilepsy were evaluated using the Ono's grading system. Molecular biological methods were used to evaluate the effects of the drugs on the levels of amino acids in the brains of each group of rats, as well as on the expression of apoptosis-related proteins. The results showed that both compounds 1 and 2 had therapeutic effects on the SD epilepsy model rats. Compound 1 showed superior performance compared to the CBD group in all indicators, indicating that compound 1 was more effective than compound 2. Furthermore, compound 1, as a derivative of cannabidiol, exhibited significant advantages in solubility through structural modification, with a 40% improvement compared to cannabidiol. This improvement not only effectively enhanced drug solubility but also increased bioavailability and stability, creating favorable conditions for drug formulation development and contributing to expanding its application potential and prospects in the pharmaceutical field.

[0184] The preferred embodiments of the present invention have been described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the scope of the technical concept of the present invention, various simple modifications can be made to the technical solution of the present invention, and these simple modifications all fall within the protection scope of the present invention.

[0185] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, the present invention will not describe the various possible combinations separately.

Claims

1. A compound or its pharmaceutically acceptable salt or stereoisomer, having the following structure: in, R1 is selected from C l-10 alkyl; X1 and X2 are independently selected from single bonds, alkylene groups, alkylene groups separated by one or more -C(=O)O- groups, alkylene groups separated by one or more -OC(=O)- groups, alkylene groups separated by one or more -OC(=O)O- groups, and alkylene groups separated by a combination of the above spacer groups; R2 and R3 are independently selected from: -H, -OH, -COOH, X3 is selected from CH2, NH, O, and S.

2. The compound according to claim 1, characterized in that, R1 is -(CH2)4CH3, X1 is -CH2CH2-, and X2 is -CH2CH2OC(=O)CH2CH2-.

3. The compound according to claim 1, characterized in that, R2 and R3 are independently selected from: -H, Preferably, both R2 and R3 are 4. The compound according to claim 1, characterized in that, The compound has the following structure: Preferably, the stereoisomers of the compound have the following structures:

5. A method for preparing the compound according to any one of claims 1-4, characterized in that, Compound I reacts with 1-1 to give compound II; Compound II undergoes transesterification to yield compound IV; Alternatively, compound II reacts with a diol, followed by transesterification, to yield compound IV; The specific synthetic route is as follows: Where R5 is C 1-6 alkyl; X4 is selected from alkylene groups.

6. The preparation method according to claim 5, characterized in that, It includes the following steps: 1) Dissolve compound I, add 1-1, a desiccant and a catalyst, and react to obtain compound II; 2) Dissolve compound II, add an alkaline reagent and a diol, react to obtain compound III; 3) Dissolve compound III, add 1-2, alkaline reagent, condensing agent and catalyst, react to obtain compound IV; The terms 1-2 are: R4 is selected from: -H, -OH, -COOH, X3 is selected from CH2, NH, O, and S.

7. The preparation method according to claim 6, characterized in that, The catalyst in step 1) is one of boron trifluoride diethyl ether, aluminum trichloride, aluminum tribromide, ferric chloride, antimony pentachloride, hydrofluoric acid, or phosphorus pentoxide, preferably boron trifluoride diethyl ether; Preferably, the molar ratio of I to 1-1 is 1:0.5 to 1:1.5; Preferably, the molar ratio of I to the catalyst is 1:0.01 to 1:0.15; Preferably, the reaction temperature in step 1) is -40 to -10°C, and the reaction time is 0.5 to 6 hours. Preferably, the reaction in step 2) is carried out under negative pressure, wherein the pressure of the negative pressure environment is -0.9 to -0.1 bar; Preferably, the reaction temperature in step 2) is 90–140°C, and the reaction time is 1–6 h.

8. The preparation method according to claim 6, characterized in that, The catalyst mentioned in step 3) is at least one of N-hydroxysuccinimide (HOSu), N-hydroxy-7-azabenzotriazole (HOAt), 1-hydroxybenzotriazole (HOBt) or 4-dimethylaminopyridine (DMAP); Preferably, the condensing agent is at least one selected from N,N'-dicyclohexylcarbodiimide (DCC), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI), N,N'-carbonyldiimidazole (CDI), N,N'-diisopropylcarbodiimide (DIC), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (HATU), O-benzotriazole-tetramethylurea hexafluorophosphate (HBTU), propyl tricyclic phosphate anhydride (T3P), n-butyl phosphate anhydride (T4P), diphenyl azidophosphate (DPPA), or bis(2-oxo-3-oxazolyl)hypophosphine chloride (BOP-Cl). Preferably, the molar ratio of Ⅲ to 1-2 is 1:1.0 to 1:5.0; Preferably, the molar ratio of III to the alkaline reagent is 1:2.0 to 1:8.0; Preferably, the molar ratio of III to the condensing agent is 1:1.5 to 1:4.5; Preferably, the molar ratio of III to the catalyst is 1:0.01 to 1:0.

15.

9. The use of a compound as described in any one of claims 1-4, or a pharmaceutically acceptable salt or stereoisomer thereof, in the preparation of a medicament for treating epilepsy.

10. A pharmaceutical composition, characterized in that, The pharmaceutical composition comprises any one of the compounds of claims 1-4 or their pharmaceutically acceptable salts, stereoisomers, and one or more pharmaceutically acceptable excipients.

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