Triazoles for protecting brain tissue and methods of making the same

Abstract

The application provides a triazole compound for protecting brain tissue and a preparation method thereof, and belongs to the technical field of biological medicines, and provides a triazole compound as shown in formula (I) and chiral isomers, enantiomers, diastereoisomers, geometric isomers, free forms or pharmaceutically acceptable salts, hydrates, solvates or esters thereof; and also provides a preparation method thereof; compared with the prior art, the application combines the internal ester ring-opening product of butylphthalide and ligustrazine through triazole to synthesize a novel triazole compound which can be used for protecting brain tissue; the preparation method of the triazole compound is simple and the raw materials are easy to obtain; the triazole compound has better biological activity and brain tissue protection effect than butylphthalide / ligustrazine, and has significant protection activity on a SH-SY5Y cell damage model.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, specifically relating to a triazole compound for protecting brain tissue and its preparation method. Background Technology

[0002] Drug therapy is one of the important methods of stroke treatment. At present, clinical drug therapy mainly relieves symptoms by providing drugs with effects such as lowering blood pressure, inhibiting platelet aggregation, and anticoagulation. However, the treatment effect is not significant and cannot effectively prevent the occurrence and recurrence of stroke. At the same time, there is currently no specific drug that can completely cure stroke. It can only alleviate the condition through a series of treatments, and the effect of drug therapy on stroke is limited.

[0003] Therefore, drug treatment for stroke has always been an important research topic for those skilled in the art. n-Butylphthalide (NBP), commonly known as celery oleoresin, is a major component of celery volatile oil. Researchers at the Institute of Materia Medica, Chinese Academy of Medical Sciences, isolated the levorotatory isomer of butylphthalide from the volatile oil of celery seeds. It is a chemical drug primarily indicated for the treatment of ischemic stroke, encompassing the mechanisms of action of both improving cerebral blood flow and protecting brain tissue, thus offering unique therapeutic effects for patients with ischemic stroke. Although butylphthalide can treat multiple pathological aspects of cerebral ischemia, its poor water solubility results in a generally low overall efficacy.

[0004] Patent application CN106928155A, based on butylphthalide, designed and prepared three compounds combining tetramethylpyrazine and butylphthalide. Tetramethylpyrazine (TMP) is an alkaloid extracted from the umbelliferous plant Ligusticum chuanxiong and is one of the effective components for treating cardiovascular and cerebrovascular diseases. Pharmacological studies have shown that tetramethylpyrazine has the effects of dilating blood vessels, increasing arterial blood flow, inhibiting platelet aggregation, and reducing platelet activity. It is characterized by complete absorption and wide distribution in vivo. However, due to its poor lipid solubility, rapid metabolism, and short half-life, tetramethylpyrazine requires frequent administration in clinical practice to maintain an effective therapeutic concentration, which can easily lead to accumulation and toxicity, thus limiting its application.

[0005] Invention patent CN106928155B discloses a tetramethylpyrazine-butylphthalide compound, its preparation method, and its application in pharmaceuticals. This tetramethylpyrazine-butylphthalide compound has the following general structural formula I: The invention uses phthalic anhydride and tetramethylpyrazine as starting materials, and prepares the tetramethylpyrazine-butylphthalide compound through bromination, nucleophilic addition, catalytic dehydration, ester hydrolysis, reduction, and esterification reactions. The pharmaceutical composition consists of the tetramethylpyrazine-butylphthalide compound as the active pharmaceutical ingredient, and pharmaceutically acceptable carriers, excipients, diluents, adjuvants, mediators, or combinations thereof. The tetramethylpyrazine-butylphthalide compound prepared by this invention has a good inhibitory effect on platelet aggregation induced by adenosine diphosphate (ADP) in vitro and has good in vivo pharmacokinetic properties, and can be used for the prevention and treatment of cardiovascular and cerebrovascular diseases and their complications.

[0006] In implementing the inventive embodiments, the inventors discovered at least the following deficiencies in the prior art:

[0007] This invention primarily employs the Born turbidimetric method to test the inhibitory activity of the target compound on ADP-induced platelet aggregation in rabbits to evaluate the compound's antiplatelet aggregation inhibitory activity. However, currently, there are many drugs available for antiplatelet aggregation, and none have significantly highlighted the unique therapeutic and protective effects of butylphthalide itself for stroke. Therefore, to fully utilize the unique effects of butylphthalide in the treatment and prevention of ischemic stroke, further development of compounds with high brain tissue protective activity based on butylphthalide is still needed for effective prevention and treatment of stroke. Summary of the Invention

[0008] This invention addresses the lack of a highly effective compound for protecting brain tissue in existing technologies. It further introduces a triazole ring and a series of modifying groups onto butylphthalide and tetramethylpyrazine, while also modifying their chiral structure to improve their pharmacological activity. The invention provides a triazole compound for protecting brain tissue and its preparation method.

[0009] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0010] The abbreviations used in this invention and their meanings are as follows:

[0011] In this invention, NBP refers to n-butylphthalide;

[0012] In this invention, DMAP refers to 4-dimethylaminopyridine;

[0013] In this invention, Et3N refers to triethylamine;

[0014] In this invention, TMP refers to tetramethylpyrazine;

[0015] In this invention, TBTU refers to O-benzotriazole-N,N,N',N'-tetramethylurea tetrafluoroboric acid;

[0016] In this invention, DIPEA refers to N,N-diisopropylethylamine;

[0017] In this invention, BPO refers to benzoyl peroxide.

[0018] In this invention, NBS refers to N-bromosuccinic anhydride.

[0019] CuTC is cuprous thiophene-2-carboxylate (I).

[0020] In a first aspect, the present invention provides a triazole compound for protecting brain tissue, the general formula of which is shown in structural formula (I):

[0021]

[0022] Where R is independently selected X is independently selected from H or halogens.

[0023] Preferably, the structural formula (Ⅰ) is any one of the following:

[0024]

[0025]

[0026] Preferably, R is selected from X is selected from H; R is selected from X is selected from H; or R is selected from H; or X is selected from H; or R is selected from X is selected from H; or R is selected from... X is selected from H;

[0027] Structural formula (Ⅰ) can be any of the following:

[0028]

[0029] More preferably, R is selected from R selected from X is selected from H; or R is selected from... X is selected from H.

[0030] Structural formula (Ⅰ) can be any of the following:

[0031]

[0032] Secondly, the present invention also provides a method for preparing a triazole compound for protecting brain tissue as shown in formula (I), comprising the following steps:

[0033] (1) Butylphthalide was hydrolyzed with alkaline solution, pH was adjusted, the solution was concentrated, and chloroacetyl chloride, 4-dimethylaminopyridine and triethylamine were added to react and intermediate 1 was obtained.

[0034] (2) Dissolve intermediate 1 obtained in step (1) with propargylamine, add O-benzotriazole-N,N,N',N'-tetramethylurea tetrafluoroboric acid and N,N-diisopropylethylamine to react and obtain intermediate 2;

[0035] (3) Ligustrazine was brominated with N-bromosuccinic anhydride using benzoyl peroxide as an initiator to obtain intermediate 3;

[0036] (4) Dissolve intermediate 3 obtained in step (3), and then add sodium azide to react and obtain intermediate 4;

[0037] (5) Dissolve intermediate 2 obtained in step (2) and intermediate 4 obtained in step (4), add catalyst to react and obtain intermediate 5;

[0038] (6) The intermediate 5 obtained in step (5) undergoes a substitution reaction with the substituted compound under the action of a catalyst to obtain the triazole compound.

[0039] Preferably, the alkali used in step (1) is selected from at least one of sodium hydroxide, potassium hydroxide and calcium hydroxide.

[0040] Preferably, the hydrolysis conditions in step (1) are: heat preservation at 55-65℃ for 1.5-3 hours, or microwave at 55-65℃ for 30-45 minutes.

[0041] Preferably, the target pH for adjusting pH in step (1) is 3-4, and the reagent used is selected from one or more of dilute hydrochloric acid, dilute sulfuric acid and dilute nitric acid.

[0042] Preferably, the concentration method in step (1) is extraction by adding an extractant; the extractant is selected from one or both of ethyl acetate and diethyl ether.

[0043] Preferably, the molar ratio of chloroacetyl chloride, 4-dimethylaminopyridine and triethylamine to butylphthalide in step (1) is 1.4-1.6:0.08-0.12:1.4-1.6:1.

[0044] Most preferably, the molar ratio of chloroacetyl chloride, 4-dimethylaminopyridine and triethylamine to butylphthalide in step (1) is 1.5:0.1:1.5:1.

[0045] Preferably, the solvent used for dissolution in step (2) includes carbon dichloride.

[0046] Preferably, the molar ratio of propargylamine to intermediate 1 in step (2) is 1.1-1.3:1.

[0047] Most preferably, the molar ratio of propargylamine to intermediate 1 in step (2) is 1.2:1.

[0048] Preferably, in step (2), the molar ratio of O-benzotriazole-N,N,N',N'-tetramethylurea tetrafluoroboric acid, N,N-diisopropylethylamine and intermediate 1 is 1.1-1.3:1.8-2.2:1.

[0049] Most preferably, the molar ratio of O-benzotriazole-N,N,N',N'-tetramethylurea tetrafluoroboric acid, N,N-diisopropylethylamine and intermediate 1 in step (2) is 1.2:2:1.

[0050] Preferably, the reaction in step (2) is carried out at a temperature of 20-30°C for 5.5-7 hours.

[0051] Most preferably, the reaction in step (2) is carried out at a temperature of 25°C for 6 hours.

[0052] Preferably, the molar ratio of benzoyl peroxide to tetramethylpyrazine in step (3) is 0.08-0.12:1.

[0053] Most preferably, the molar ratio of benzoyl peroxide to tetramethylpyrazine in step (3) is 0.1:1.

[0054] Preferably, the molar ratio of N-bromosuccinic anhydride to tetramethylpyrazine in step (3) is 0.25-0.35:1.

[0055] Most preferably, the molar ratio of N-bromosuccinic anhydride to tetramethylpyrazine in step (3) is 0.3:1.

[0056] Preferably, the bromination reaction in step (3) is carried out by reflux.

[0057] More preferably, the reflux temperature is 75-85°C and the time is 3.5-5 hours.

[0058] Preferably, the solvent used for dissolution in step (4) includes acetonitrile.

[0059] Preferably, the molar ratio of sodium azide to intermediate 3 in step (4) is 1.15-1.25:1.

[0060] Most preferably, the molar ratio of sodium azide to intermediate 3 in step (4) is 1.2:1.

[0061] Preferably, the reaction conditions in step (4) are microwaved at 55-65°C for 55-70 minutes.

[0062] Most preferably, the reaction conditions in step (4) are microwave at 60°C for 60 min.

[0063] Preferably, the molar ratio of intermediate 2 to intermediate 4 in step (5) is 1:0.9-1.1.

[0064] Most preferably, the molar ratio of intermediate 2 to intermediate 4 in step (5) is 1:1.

[0065] Preferably, the solvent used for dissolution in step (5) includes dichloromethane.

[0066] Preferably, the catalyst in step (5) comprises cuprous thiophene-2-carboxylate (I).

[0067] Preferably, the reaction temperature in step (5) is 20-30°C.

[0068] Preferably, the catalyst in step (6) includes cesium carbonate; the molar ratio of cesium carbonate to intermediate 5 is 1.4-1.6:1.

[0069] Preferably, the molar ratio of the substituted compound to intermediate 5 in step (6) is 1.4-1.6:1.

[0070] More preferably, the substituted compound in step (6) is selected from morpholine, N-methylpiperazine, piperazine, diethylamine and dimethylamine.

[0071] Preferably, the substitution reaction in step (6) is carried out at 60-70°C for 100-140 min; or at 60-70°C under microwave for 50-65 min.

[0072] Most preferably, the substitution reaction in step (6) is carried out at 65°C for 120 min; or at 65°C under microwave for 60 min.

[0073] Thirdly, the present invention also provides chiral isomers, enantiomers, diastereomers, geometric isomers, free forms, or pharmaceutically acceptable salts, hydrates, solvates, or esters of triazole compounds as shown in formula (I) for protecting brain tissue.

[0074] Preferably, the general formula of the chiral isomer is shown in structural formula S-(Ⅰ):

[0075]

[0076] Where R is independently selected X is independently selected from H or halogens.

[0077] Fourthly, the present invention also provides the use of triazole compounds of formula (I) for protecting brain tissue, their chiral isomers, enantiomers, diastereomers, geometric isomers, free forms, or pharmaceutically acceptable salts, hydrates, solvates, or esters in the preparation of medicaments; said medicaments are used to prevent and / or treat the following human diseases: stroke, Alzheimer's disease, epilepsy, and ALS.

[0078] Preferably, the general formula of the chiral isomer is shown in structural formula S-(Ⅰ).

[0079] Preferably, the drug is used for the prevention and / or treatment of ischemic stroke.

[0080] Fifthly, the present invention also provides a pharmaceutical composition comprising a triazole compound of formula (I) for protecting brain tissue, a chiral isomer, an enantiomer, a diastereomer, a geometric isomer, a free form or a pharmaceutically acceptable salt, hydrate, solvate or ester thereof.

[0081] Preferably, the pharmaceutical composition further comprises pharmaceutically acceptable excipients, said excipients comprising at least one of the following: binders, fillers, diluents, disintegrants, suspending agents, suspending aids, sustained-release agents, lyophilization protectants, coating agents, enteric materials, lubricants, flow aids, anti-adhesion agents, sweeteners, flavoring agents, plasticizers, opacifiers, solubilizers, humectants, solvents, osmotic pressure regulators, colorants, pigments, surfactants, emulsifiers, water-soluble matrices, fat-soluble matrices, lipophilic matrices, pore-forming agents, gelling agents, preservatives, buffers, chelating agents, and antioxidants.

[0082] Preferably, the general formula of the chiral isomer is shown in structural formula S-(Ⅰ).

[0083] In a sixth aspect, the present invention also provides the use of the above-described pharmaceutical composition in the preparation of a medicament for the prevention and / or treatment of the following human diseases: stroke, Alzheimer's disease, epilepsy, and ALS.

[0084] Preferably, the drug is used for the prevention and / or treatment of ischemic stroke.

[0085] Compared with the prior art, the present invention has the following beneficial effects:

[0086] This invention synthesizes a novel triazole compound that can be used to protect brain tissue by combining the lactone ring-opening product of butylphthalide with tetramethylpyrazine via triazole.

[0087] The triazole compound preparation method provided by this invention uses readily available raw materials and has a simple synthetic route; it has better biological activity and brain tissue protection effect than butylphthalide / tetramethylamine, and has significant protective activity against SH-SY5Y cell injury model. Detailed Implementation

[0088] It is worth noting that the raw materials used in this invention are all commercially available products, and their sources are not specifically limited.

[0089] The following sources of raw materials are provided as examples:

[0090] The reagents used in this invention, namely NBP, sodium iodide, HCl, HBr, chloroacetyl chloride, triethylamine, DMAP, methanol, potassium carbonate, cesium carbonate, acetone, tetrahydrofuran, TMP, NBS, BPO, carbon tetrachloride, TBTU, DIPEA, morpholine, N-methylpiperazine, diethylamine, dimethylamine, and 1-Boc-piperazine, were purchased from Energy Chemical Company and are all commercially available reagents of analytical or chemical purity.

[0091] The instruments and equipment used in this invention:

[0092] IKA RCT basic magnetic stirrer [IKA Instrument Equipment Co., Ltd.]

[0093] Mettler Toledo AL104 electronic balance [Mettler Toledo (Beijing) Precision Instruments Co., Ltd.];

[0094] Bruker Avance II 600 nuclear magnetic resonance spectrometer [Bruker (Beijing) Technology Co., Ltd.]

[0095] Thermo Fisher Scientific LTQ-Obitrap XL LC-MS / MS (Thermo Fisher Scientific).

[0096] The present invention will be further described below with reference to the embodiments.

[0097] Examples of triazole compounds for protecting brain tissue and their preparation methods

[0098] Examples 1-20 of this invention provide triazole compound B1-20 for protecting brain tissue and its preparation method, as detailed below:

[0099] Preparation of Intermediate 1

[0100] The structural formulas of intermediate 1 and its chiral isomer (S) are as follows:

[0101]

[0102] Compound NBP (10.0 g, 52.6 mmol) was added to 60 mL of an ethanol-water mixture (2:1 v / v), followed by potassium hydroxide (4.4 g, 78.4 mmol). The mixture was microwaved at 60 °C for 40 min. After the reaction, the solvent was evaporated to dryness. The concentrate was diluted with water, and the pH was adjusted to 3-4 with dilute hydrochloric acid, resulting in the precipitation of a white solid. The solid was extracted three times with ethyl acetate, and the organic layers were combined and dried over anhydrous magnesium sulfate. Chloroacetyl chloride (6.3 mL, 78.9 mmol) was added to the ethyl acetate solution of intermediate 1, followed by the addition of DMAP (642.3 mg, 5.26 mmol) and then triethylamine (11 mL, 78.9 mmol). The mixture was stirred and reacted. After the reaction, the solid was extracted with water, and the organic layer was collected, dried over anhydrous sodium sulfate, filtered, and the solvent was evaporated to dryness to obtain the crude product. The crude product was purified by silica gel column chromatography with a V (petroleum ether):V (ethyl acetate) ratio of 10:1 to obtain 7.6 g of a pale yellow solid, with a yield of 51%. This crude product was used in the next reaction.

[0103] Preparation of intermediate 2

[0104] The structural formulas of intermediate 2 and its chiral isomer (S) are as follows:

[0105]

[0106] Intermediate 1 (5 g, 17.6 mmol) was dissolved in 30 mL of dichloromethane, followed by the addition of TBTU (6.67 g, 29.6 mmol) and DIPEA (5.8 mL, 44.2 mmol). After the reaction was complete, the mixture was extracted three times with brine, and the organic layer was collected. After removing water, the solvent was evaporated to dryness, yielding the crude product. The crude product was purified by silica gel column chromatography with a ratio of V (petroleum ether):V (ethyl acetate) of 10:1 to give 3.53 g of a pale yellow solid, with a yield of 62.3%. This was used for the next reaction.

[0107] Intermediate 2 1 H NMR, 13 The C NMR and HRMS data are shown below:

[0108] 1 H-NMR (600MHz, CDCl3) δ: 8.09 (dd, J=8.0Hz, 1.1Hz, 1H, H-3), 7.61-7.58 (td, J=8.1Hz, 1.4Hz, 1H, H-5), 7.58-7.56 (dd, J=8 .1Hz,1.5Hz,1H,H-6),7.39(td,1H,J=7.4Hz,1.5Hz,H-4),6.77(m,1H,C(CH)O),4.10(d,2H,CO(CH2)Cl),0.91(t,3H,CH3);13 C-NMR (150)

[0109] MHz, CDCl3)δ:172.0,166.7,143.7,133.5,131.5,127.7,126.9,126.1,75..0,41.1,36.5,27.9,22.3,14.0; HRMS:Calcd.for C 14 H 17 ClO4(M+Na):307.0708.Found:307.0710

[0110] Preparation of intermediate 3

[0111] The structural formulas of intermediate 3 and its chiral isomer (S) are as follows:

[0112]

[0113] Compound TMP (5 g, 36.7 mmol) was dissolved in carbon tetrachloride (50 mL), followed by the addition of NBS (2.179 g, 12.2 mmol) and BPO (890 mg, 3.6 mmol). The mixture was then microwaved at 65 °C for 1 h. After the reaction was complete, the mixture was filtered, the solvent was evaporated to dryness, and the solution was purified by silica gel column chromatography at a ratio of V (petroleum ether):V (ethyl acetate) of 20:1 to give 1.83 g of a pale white solid, with a yield of 69%. This solid was used in the next reaction.

[0114] Preparation of intermediate 4

[0115] The structural formulas of intermediate 4 and its chiral isomer (S) are as follows:

[0116]

[0117] Dissolve intermediate 3 (500 mg, 2.32 mmol) in acetonitrile (2 ml), then add NaN3, and microwave at 60 °C for 1 h. After the reaction is complete, filter under vacuum, evaporate the solvent to dryness, and use it directly in the next reaction without further processing.

[0118] Preparation of intermediate 5

[0119] The structural formulas of intermediate 5 and its chiral isomer (S) are as follows:

[0120]

[0121] Accurately weigh 2 g (7 mmol) of intermediate 2, dissolve it in 5 mL of dichloromethane, and add intermediate 4 (1.16 g, 7 mmol) and Cutc (260 mg, 7 mmol) sequentially at room temperature. Stir for 2 h at room temperature. After the reaction is complete as detected by TLC, filter and concentrate to obtain the crude product. Purify the crude product by silica gel column chromatography with a V(dichloromethane):V(methanol) ratio of 15:1 to obtain 1.8 g of a pale yellow solid, with a yield of 56%. Used for the next reaction.

[0122] Intermediate 5 1 H NMR, 13 The C NMR and HRMS data are shown below.

[0123] 1 H-NMR(600MHz, CDCl3)δ:7.72(s,1H,CCHN),7.45-7.42(m,1H,H-3),7.42-7.41(m,1H,H-5),7.41-7.40(m,1H,H-6),7.33-7.29( m,1H,H-4),7.20(m,1H,CONH),6.05(m,1H,C(CH)O),5.61(m,2H,NCH2C),4.74-4.66(m,2H,COCH2Cl),4.04(d,J=14.8Hz,2H,NHC H 2),2.54(s,3H,CH3),2.51(s,3H,CH3),2.50(s,3H,CH3),0.86(t,J=7.2Hz,3H,CH3); 13 C-NMR (150MHz, CDCl3) δ:169.0,167.3,152.1,149.6,148.9,144.8,143.6,138.4,135.3,130.6 ,128.2,127.3,126.2,122.5,75.9,53.2,41.0,36.4,35.6,27.6,22.4,21.7,21.5,20.7,13.9.

[0124] HRMS:Calcd.for C 25 H 31 ClN6O3(M+Na):521.2038.Found:521.2038.

[0125] Example 1: Preparation of compound B1

[0126] The structural formulas of compound B1 and its levorotatory isomer, compound B1(S), are as follows:

[0127]

[0128] Preparation method: Accurately weigh (1 g, 1.6 mmol) of intermediate 5 or intermediate 5(S) into a microwave tube, dissolve it in 3 ml of tetrahydrofuran, and then add cesium carbonate (0.9 g, 3.8 mmol) and morpholine (0.24 ml, 3.8 mmol) sequentially. Microwave reaction is carried out at 65 °C for 1 h. After the reaction is complete as detected by TLC, the crude product is obtained by filtration and concentration. Purification by silica gel column chromatography (V:V:ethyl acetate = 1:4) yields 0.43 g of a yellow viscous liquid, with a yield of 39%. Compound B1 and its levorotatory isomer B1(S) are also discussed. 1 H NMR, 13 CNMR, HRMS, and specific rotation values ​​are shown below.

[0129] (4-iodo-2-(((1-((3,5,6-trimethylpyrazin-2-yl)methyl)-1H-1,2,3-triazol-4-yl)methyl)carbamoyl)phenyl)pentyl 2-morpholinoacetate(TY-LY-NBP-74):Pale yellow oily; 72% yield; -28.2 (MeOH); 1 H-NMR(600MHz, CDCl3)δ:7.67(d,J=1.9Hz,1H,H-3),,7.65-7.63(m,1H,H-5),7.64(s,1H,CCHN),7.60(m, 1H,CONH),7.03(d,J=8.4Hz,1H,H-6),5.79(m,1H,C(CH)O),5.54(d,J=14.5Hz,2H,NCH2C),4.59(m,2H,NHC H 2),3.63(t,J=4.5Hz,4H,CH2OCH2),3.12(d,J=16.7Hz,2H,COCH2N),2.48(s,3H,CH3),2.47-2. 44,2.42-2.38(m,4H,CH2NCH2),2.44(s,3H,CH3),2.43(s,3H,CH3),0.76(t,J=7.3Hz,3H,CH3); 13C-NMR (150MHz, CDCl3) δ:169.5,166.4,151.0,148.6,147.8,143.6,142.5,138.3,137.2,136.2,135.3,126.8,,12 1.5,92.3,73.1,65.6(2C),58.5,52.2(2C),52.1,35.2,34.5,26.4,21.2,20.7,20.5,19.6,12.8.; HRMS:Calcd.for C 29 H 38 IN7O4(M+H):676.2103.Found:676.2103.

[0130] Example 2: Preparation of compound B2

[0131] The structural formulas of compound B2 and its levorotatory isomer B2(S) are as follows:

[0132]

[0133] Preparation method: Referring to Example 1, only 3-iodo-butylphthalide was replaced with 3-bromo-butylphthalide. The product was a yellow viscous liquid with a yield of 43%. Compound B2 and its levorotatory isomer B2(S) were also discussed. 1 H NMR, 13 The C NMR, HRMS, and specific rotation data are shown below.

[0134] -31.7 (MeOH); 1 H-NMR (600MHz, CDCl3) δ: 7.64 (s, 1H, CCHN), 7.63-7.60 (m, 1H, CONH), 7.48 (d, J = 2.1Hz, 1H, H-3), 7.44 (dd, J = 8.5H z,2.1Hz,1H,H-5),7.17(d,J=8.4Hz,1H,H-6),5.80(m,1H,C(CH)O),5.54(d,J=14.5Hz,2H,NCH2C),4.60(m,2H,NHC H 2),3.63(t,J=4.6Hz,4H,CH2OCH2),3.11(d,J=16.7Hz,2H,COCH2N),2.48(s,3H,CH3),2.47-2. 44,2.42-2.38(m,4H,CH2NCH2),2.44(s,3H,CH3),2.43(s,3H,CH3),0.76(t,J=7.2Hz,3H,CH3); 13C-NMR (150MHz, CDCl3) δ:169.6,166.5,151.1,148.6,147.8,143.6,142.5,136.6,136.1,132.4,129.6,12 6.8,121.5,120.9,73.0,65.6(2C),58.6,52.3(2C),52.1,35.2,34.5,26.4,21.2,20.7,20.5,19.6,12.8.

[0135] HRMS:Calcd.for C 29 H 38 BrN7O4(M+H):628.2242.Found:628.2242.

[0136] Example 3: Preparation of compound B3

[0137] The structural formulas of compound B3 and its levorotatory isomer, compound B3(S), are as follows:

[0138]

[0139] Preparation method: Referring to Example 1, only 3-iodo-butylphthalide was replaced with 3-chloro-butylphthalide. The product was a yellow viscous liquid with a yield of 66%. Compound B3 and its levorotatory isomer B3(S) 1 H NMR, 13 The C NMR, HRMS, and specific rotation values ​​are shown below.

[0140] -27.1 (MeOH); 1 H-NMR (600MHz, CDCl3) δ: 7.65 (s, 1H, CCHN), 7.64-7.62 (m, 1H, CONH), 7.33 (d, J = 2.1Hz, 1H, H-3), 7.29 (dd, J = 8.4H z,2.1Hz,1H,H-5),7.24(d,J=8.5Hz,1H,H-6),5.82(m,1H,C(CH)O),5.54(d,J=14.5Hz,2H,NCH2C),4.60(m,2H,NHC H 2),3.64(t,J=4.5Hz,4H,CH2OCH2),3.15(d,J=16.6Hz,2H,COCH2N),2.48(s,3H,CH3),2.52-2. 48,2.47-2.44(m,4H,CH2NCH2),2.44(s,3H,CH3),2.43(s,3H,CH3),0.76(t,J=7.1Hz,3H,CH3); 13C-NMR (150MHz, CDCl3) δ:169.6,166.7,151.1,148.6,147.8,143.6,142.5,136.0,135.9,132.9,129.5,12 6.7,126.6,121.5,73.0,65.7(2C),58.6,52.3(2C),52.1,35.3,34.5,26.4,21.2,20.7,20.5,19.6,12.8.

[0141] HRMS:Calcd.for C 29 H 38 ClN7O4(M+H):584.2746.Found::584.2746.

[0142] Example 4: Preparation of compound B4 and its levorotatory isomer, compound B4(S)

[0143]

[0144] Referring to Example 1, only 3-iodo-butylphthalide was replaced with butylphthalide; the product was a yellow viscous liquid with a yield of 39%. Compound B4 and its levorotatory isomer, B4(S), were also discussed. 1 H NMR, 13 The C NMR, HRMS, and specific rotation values ​​are shown below. -24.8 (MeOH); 1 H-NMR(600MHz, CDCl3)δ:7.65(s,1H,CCHN),7.54(m,1H,CONH),7.35-7.33(m,1H,H-3),7.33-7.31(m,1H,H-5),7. 31-7.29(m,1H,H-6),7.25-7.21(m,1H,H-4),5.87(m,1H,C(CH)O),5.53(d,J=14.6Hz,2H,NCH2C),4.62(m,2H,NHC H 2),3.62(t,J=4.6Hz,4H,CH2OCH2),3.12(d,J=16.6Hz,2H,COCH2N),2.47(s,3H,CH3),2.47-2. 44,2.42-2.37(m,4H,CH2NCH2),2.44(s,3H,CH3),2.43(s,3H,CH3),0.77(t,J=7.2Hz,3H,CH3); 13C-NMR (150MHz, CDCl3) δ:169.5,168.2,151.0,148.5,147.9,143.9,142.5,137.6,134.4,129.3,127.0,126.6,125 .0,121.5,73.5,65.7(2C),58.6,52.2(2C),52.1,35.4,34.5,26.5,21.3,20.7,20.5,19.6,12.9.; HRMS:Calcd.for C 29 H 39 N7O4(M+H):550.3135.Found:550.3135.

[0145] Example 5: Preparation of compound B5

[0146] The structural formulas of compound B5 and its levorotatory isomer, compound B5(S), are as follows:

[0147]

[0148] Referring to Example 1, only morpholine was replaced with N-methylpiperazine; the product was a yellow viscous liquid with a yield of 64%. Compound B5 and its levorotatory isomer B5(S) were also discussed. 1 H NMR, 13 The C NMR, HRMS, and specific rotation values ​​are shown below.

[0149] -27.8 (MeOH); 1 H-NMR (600MHz, CDCl3) δ: 7.67 (d, J=1.8Hz, 1H, H-3), 7.65-7.63 (m, 1H, H-5), 7.65 (s, 1H, CCHN), 7.63-7.60 ( m,1H,CONH),7.03(d,J=8.3Hz,1H,H-6),5.78(m,1H,C(CH)O),5.54(d,J=14.5Hz,2H,NCH2C),4.59(m,2H,NHC H 2),3.15(d,J=16.9Hz,2H,COCH2N),2.80-2.50(m,8H,(CH2NCH2)2),2.48(s,3H,CH3 ),2.44(s,3H,CH3),2.43(s,3H,CH3),2.36(s,3H,NCH3),0.76(t,J=7.1Hz,3H,CH3); 13C-NMR (150MHz, CDCl3) δ:169.5,166.4,151.1,148.6,147.8,143.6,142.5,138.4,137.1,136.2,135.3,1 26.9,,121.6,92.4,73.2,57.9,53.2,52.1,50.7,44.0,35.2,34.5,26.4,21.2,20.7,20.5,19.7,12.8.;

[0150] HRMS:Calcd.for C 30 H 41 IN8O3(M+H):689.2418.Found:689.2418.

[0151] Example 6: Preparation of compound B6

[0152] The structural formulas of compound B6 and its levorotatory isomer, compound B6(S), are as follows:

[0153]

[0154] Referring to Example 1, only 3-iodobutylphthalide was replaced with 3-bromobutylphthalide, and morpholine was replaced with N-methylpiperazine. The product was a yellow viscous liquid with a yield of 45%. Compound B6 and its levorotatory isomer B6(S) were also discussed. 1 H NMR, 13 The C NMR, HRMS, and specific rotation values ​​are shown below.

[0155] -35.3 (MeOH); 1 H-NMR(600MHz, CDCl3)δ:7.69-7.65(m,1H,CONH),7.65(s,1H,CCHN),7.48(d,J=1.9Hz,1H,H-3),7.45(m, 1H,H-5),7.18(d,J=8.4Hz,1H,H-6),5.79(m,1H,C(CH)O),5.54(d,J=14.5Hz,2H,NCH2C),4.60(m,2H,NHC H 2),3.15(d,J=16.6Hz,2H,COCH2N),2.90-2.49(m,8H,(CH2NCH2)2),2.48(s,3H,CH3 ),2.44(s,3H,CH3),2.43(s,3H,CH3),2.33(s,3H,NCH3),0.76(t,J=7.2Hz,3H,CH3); 13C-NMR (150MHz, CDCl3) δ:169.5,166.5,151.0,148.6,147.8,143.6,142.5,136.5,136.1,132.4,129.5,1 26.8,121.6,,120.9,73.1,57.9,53.3,52.1,50.9,44.2,35.2,34.5,26.4,21.2,20.7,20.5,19.6,12.8.;

[0156] HRMS:Calcd.for C 30 H 41 BrN8O3(M+H):641.2558.Found:641.2558.

[0157] Example 7: Preparation of compound B7

[0158] The structural formulas of compound B7 and its levorotatory isomer, compound B7(S), are as follows:

[0159]

[0160] Preparation method: Referring to Example 1, only 3-iodo-butylphthalide was replaced with 3-chloro-butylphthalide, and morpholine was replaced with N-methylpiperazine. The product was a yellow viscous liquid with a yield of 58%. Compound B7 and its levorotatory isomer B7(S) 1 H NMR, 13 The C NMR, HRMS, and specific rotation values ​​are shown below.

[0161] -37.8 (MeOH); 1 H-NMR(600MHz, CDCl3)δ:7.69-7.66(m,1H,CONH),7.65(s,1H,CCHN),7.33(d,J=2.2Hz,1H,H-3),7.30(dd,J=8.4H z,2.3Hz,1H,H-5),7.24(d,J=8.4Hz,1H,H-6),5.81(m,1H,C(CH)O),5.54(d,J=14.6Hz,2H,NCH2C),4.60(m,2H,NHC H 2),3.16(d,J=16.9Hz,2H,COCH2N),2.76-2.49(m,8H,(CH2NCH2)2),2.48(s,3H,CH3 ),2.44(s,3H,CH3),2.43(s,3H,CH3),2.38(s,3H,NCH3),0.76(t,J=7.2Hz,3H,CH3); 13C-NMR (150MHz, CDCl3) δ:169.5,166.7,151.0,148.6,147.8,143.6,142.5,136.0,135.9,132.9,129.5,1 26.7,126.6,121.6,73.1,57.8,53.2,52.1,50.5,44.0,35.2,34.5,26.4,21.2,20.7,,20.5,19.6,12.8.;

[0162] HRMS:Calcd.for C 30 H 41 ClN8O3(M+H):597.3061.Found::597.3061.

[0163] Example 8: Preparation of compound B8

[0164] The structural formulas of compound B8 and its levorotatory isomer B8(S) are as follows:

[0165]

[0166] Referring to Example 1, only 3-iodobutylphthalide was replaced with butylphthalide and morpholine was replaced with N-methylpiperazine. The product was a yellow viscous liquid with a yield of 39%. Compound B8 and its levorotatory isomer B8(S) were also discussed. 1 H NMR, 13 The C NMR, HRMS, and specific rotation values ​​are shown below.

[0167] -24.3 (MeOH); 1 H-NMR (600MHz, CDCl3) δ: 7.66 (s, 1H, CCHN), 7.50 (m, 1H, CONH), 7.35-7.34 (m, 1H, H-3), 7.34-7.32 (m, 1H, H-5), 7. 31-7.29(m,1H,H-6),7.25-7.22(m,1H,H-4),5.89(m,1H,C(CH)O),5.54(d,J=14.5Hz,2H,NCH2C),4.60(m,2H,NHC H 2),3.22(d,J=16.9Hz,2H,COCH2N),3.08-2.64(m,8H,(CH2NCH2)2),2.62(s,3H,NCH 3),2.47(s,3H,CH3),2.44(s,3H,CH3),2.43(s,3H,CH3),0.77(t,J=7.2Hz,3H,CH3); 13C-NMR(150MHz, CDCl3)δ:169.0,168.1,151.1,148.6,147.9,143.8,142.5,137.4,134.3,129.5,127.1,, 126.6,125.0,121.6,73.8,57.2,52.7,52.1,48.8,42.8,35.3,34.4,26.5,21.3,20.7,20.5,19.7,12.9.

[0168] HRMS:Calcd.for C 30 H 42 N8O3(M+H):563.3453.Found:563.3453.

[0169] Example 9: Preparation of compound B9

[0170] The structural formulas of compound B9 and its levorotatory isomer, compound B9(S), are as follows:

[0171]

[0172]

[0173] Referring to Example 1, only morpholine was replaced with diethylamine; the product was a yellow viscous liquid with a yield of 18%. Compound B9 and its levorotatory isotropic form, B9(S), were also discussed. 1 H NMR, 13 The C NMR, HRMS, and specific rotation values ​​are shown below.

[0174] -29.8 (MeOH); 1 H-NMR (600MHz, CDCl3) δ:7.71-7.68(m,1H,CONH),7.67(d,J=1.8Hz,1H,H-3),7.66-7.63(dd,J=8.3Hz,1.9Hz,1H,H-5 ),7.64(s,1H,CCHN),7.04(d,J=8.3Hz,1H,H-6),5.77(m,1H,C(CH)O),5.53(d,J=14.6Hz,2H,NCH2C),4.60(m,2H,NHC H 2),3.24(d,J=17.0Hz,2H,COCH2N),2.52(m,4H,N(C H 2)2(CH3)2),2.48(s,3H,CH3),2.44(s,3H,CH3),2.43(s,3H,CH3),0.94(t,J=7.2Hz,3H,N(CH2)2(CH 3)2),0.76(t,J=7.1Hz,3H,CH3); 13 C-NMR (150MHz, CDCl3) δ: 166.4 (2C), 151.0, 148.6, 147.8, 143.7, 142.5, 138.3, 137.3, 136.2, 135.4, 126 .9,121.5,92.3,72.9,53.0,52.1,46.6(2C),35.3,34.5,,26.4,21.2,,20.7,20.5,19.6,12.8,10.9(2C).

[0175] HRMS:Calcd.for C 29 H 40 IN7O3(M+H):662.2310.Found:662.2310.

[0176] Example 10: Preparation of compound B10

[0177] The structural formulas of compound B10 and its levorotatory isomer, B10(S), are as follows:

[0178]

[0179] Referring to Example 1, only 3-iodo-butylphthalide was replaced with 3-bromo-butylphthalide, and morpholine was replaced with diethylamine. The product was a yellow viscous liquid with a yield of 20%. Compound B10 and its levorotatory isomer, B10(S), were also discussed. 1 H NMR, 13 The C NMR, HRMS, and specific rotation values ​​are shown below.

[0180] -33.5 (MeOH); 1 H-NMR (600MHz, CDCl3) δ: 7.80 (m, 1H, CONH), 7.71 (s, 1H, CCHN), 7.56 (d, J = 2.1Hz, 1H, H-3), 7.51 (dd, J = 8.4Hz, 2 .0Hz,1H,H-5),7.25(d,J=8.4Hz,1H,H-6),5.84(m,1H,C(CH)O),5.61(d,J=14.7Hz,2H,NCH2C),4.68(m,2H,NHC H 2),3.28(d,J=16.9Hz,2H,COCH2N),2.56(m,4H,N(C H2)2(CH3)2),2.55(s,3H,CH3),2.51(s,3H,CH3),2.50(s,3H,CH3),0.99(t,J=7.2Hz,3H,N(CH2)2(C H 3)2),0.83(t,J=7.2Hz,3H,CH3); 13 C-NMR (150MHz, CDCl3) δ:172.2,167.7,152.1,149.6,148.9,144.7,143.6,137.7,137.2,133.4,130.7,127 .9,122.6,121.8,73.8,54.3,53.1,47.7(2C),36.4,35.6,27.5,,22.3,21.7,,21.5,20.7,13.9,12.1(2C).

[0181] HRMS:Calcd.for C 29 H 40 BrN7O3(M+H):614.2449.Found:614.2449.

[0182] Example 11: Preparation of compound B11

[0183] The structural formulas of compound B11 and its levorotatory isomer, compound B11(S), are as follows:

[0184]

[0185] Referring to Example 1, only 3-iodo-butylphthalide was replaced with 3-chloro-butylphthalide, and morpholine was replaced with diethylamine. The product was a yellow viscous liquid with a yield of 20%. Compound B11 and its levorotatory isomer B11... 1 H NMR, 13 The C NMR, HRMS, and specific rotation values ​​are shown below.

[0186] -38.4 (MeOH); 1 H-NMR (600MHz, CDCl3) δ: 7.72 (m, 1H, CONH), 7.65 (s, 1H, CCHN), 7.33 (d, J = 1.7Hz, 1H, H-3), 7.30-7.26 (m, 1H,H-5),7.25(d,J=8.5Hz,1H,H-6),5.80(m,1H,C(CH)O),5.53(d,J=14.5Hz,2H,NCH2C),4.61(m,2H,NHC H 2),3.28(d,J=16.9Hz,2H,COCH2N),2.56(m,4H,N(CH 2)2(CH3)2),2.48(s,3H,CH3),2.44(s,3H,CH3),2.43(s,3H,CH3),0.95(t,J=7.1Hz,3H,N(CH2)2(C H 3)2),0.81(t,J=6.8Hz,3H,CH3); 13 C-NMR (150MHz, CDCl3) δ:170.5,166.7,151.0,148.6,147.8,143.7,142.5,136.0,135.9,132.8,129.4,1 26.7,126.6,121.5,72.8,52.8,52.1,46.5(2C),35.3,34.5,26.4,21.2,20.7,20.5,19.6,12.8,10.7(2C)

[0187] HRMS:Calcd.for C 29 H 40 ClN7O3(M+H):570.2954.Found::570.2954.

[0188] Example 12: Preparation of compound B12

[0189] The structural formulas of compound B12 and its levorotatory isomer, B12(S), are as follows:

[0190]

[0191] Referring to Example 1, only 3-iodo-butylphthalide was replaced with butylphthalide and morpholine was replaced with diethylamine. The product was a yellow viscous liquid with a yield of 28%. Compound B12 and its levorotatory isomer B12(S) were also discussed. 1 H NMR, 13 The C NMR, HRMS, and specific rotation values ​​are shown below.

[0192] -21.8 (MeOH); 1 H-NMR(600MHz, CDCl3)δ:7.66(s,1H,CCHN),7.63(m,1H,CONH),7.36-7.33(m,1H,H-3),7.33-7.32(m,1H,H-5),7. 32-7.30(m,1H,H-6),7.23-7.21(m,1H,H-4),5.85(m,1H,C(CH)O),5.53(d,J=14.5Hz,2H,NCH2C),4.62(m,2H,NHC H2),3.26(d,J=16.7Hz,2H,COCH2N),2.64(m,4H,N(C H 2)2(CH3)2),2.47(s,3H,CH3),2.43(s,3H,CH3),2.42(s,3H,CH3),0.94(t,J=7.2Hz,3H,N(CH2)2(C H 3)2),0.77(t,J=7.1Hz,3H,,CH3); 13 C-NMR(150MHz, CDCl3)δ:168.2(2C),151.0,148.5,147.9,144.0,142.6,137.6,134.4,129.3,126.9,126 .6,125.0,121.5,73.3,67.0,52.1,46.6(2C),35.5,34.5,26.6,21.3,20.7,20.5,19.6,12.9,10.9(2C).

[0193] HRMS:Calcd.for C 29 H 41 N7O3(M+H):536.3344 Found:536.3344.

[0194] Example 13: Preparation of compound B13

[0195] The structural formulas of compound B13 and its levorotatory isomer, B13(S), are as follows:

[0196]

[0197] Referring to Example 1, only morpholine was replaced with dimethylamine; the product was a yellow viscous liquid with a yield of 20%. Compound B13 and its levorotatory isomer, compound B13... 1 H NMR, 13 The C NMR, HRMS, and specific rotation values ​​are shown below.

[0198] -31.8 (MeOH); 1 H-NMR(600MHz, CDCl3)δ:7.68(d,J=1.9Hz,1H,H-3),7.67-7.65(m,1H,CONH),7.65-7.64(m,1H,H-5),7.64( s,1H,CCHN),7.05(d,J=8.4Hz,1H,H-6),5.79(m,1H,C(CH)O),5.54(d,J=14.5Hz,2H,NCH2C),4.60(m,2H,NHC H2),3.08(d,J=16.5Hz,2H,COCH2N),2.48(s,3H,CH3),2.44(s,3H,CH3),2.43(s,3H,CH3),2.21(s,6H,N(CH3)2,0.76(t,J=7.1Hz,3H,CH3); 13 C-NMR (150MHz, CDCl3) δ:170.0,166.5,151.0,148.6,147.9,143.7,142.5,138.3,137.2,136.3,135 .4,126.9,121.5,92.3,73.1,59.3,52..1,44.2(2C),35.2,34.5,26.4,21.2,20.7,20.5,19.6,12.8.

[0199] HRMS:Calcd.for C 27 H 36 IN7O3(M+H):634.1997.Found:634.1997.

[0200] Example 14: Preparation of compound B14

[0201] The structural formulas of compound B14 and its levorotatory isomer, B14(S), are as follows:

[0202]

[0203] Referring to Example 1, only 3-iodo-butylphthalide was replaced with 3-bromo-butylphthalide, and morpholine was replaced with dimethylamine. The product was a yellow viscous liquid with a yield of 32%. Compound B14 and its levorotatory isomer B14(S) were also discussed. 1 H NMR, 13 The C NMR, HRMS, and specific rotation values ​​are shown below.

[0204] -33.8 (MeOH) 1 H-NMR (600MHz, CDCl3) δ: 7.80 (m, 1H, CONH), 7.72 (s, 1H, CCHN), 7.56 (d, J = 2.1Hz, 1H, H-3), 7.52 (dd, J = 8.5Hz, 2 .3Hz,1H,H-5),7.27(d,J=8.5Hz,1H,H-6),5.88(m,1H,C(CH)O),5.61(d,J=14.5Hz,2H,NCH2C),4.68(m,2H,NHC H2),3.14(d,J=16.5Hz,2H,COCH2N),2.55(s,3H,CH3),2.51(s,3H,CH3),2.50(s,3H,CH3),2.26(s,6H,N(CH3)2,0.83(t,J=7.2Hz,3H,CH3); 13 C-NMR (150MHz, CDCl3) δ:171.2,167.6,152.1,149.6,148.9,144.7,143.6,137.6,137.3,133.4,130 .6,128.0,122.6,121.9,74.0,60.5,53.1,45.3(2C),36.2,35.6,27.5,22.3,21.7,21.5,20.7,13.9.

[0205] HRMS:Calcd.for C 27 H 36 BrN7O3(M+H):586.2131.Found:586.2131.

[0206] Example 15: Preparation of compound B15

[0207] The structural formulas of compound B15 and its levorotatory isomer B15(S) are as follows:

[0208]

[0209] Referring to Example 1, only 3-iodo-butylphthalide was replaced with 3-chloro-butylphthalide, and morpholine was replaced with dimethylamine. The product was a yellow viscous liquid with a yield of 30%. Compound B15 and its levorotatory isomer B15(S) were also discussed. 1 H NMR, 13 The C NMR, HRMS, and specific rotation values ​​are shown below.

[0210] -40.1 (MeOH) 1 H-NMR(600MHz,CDCl3)δ:7.70(m,1H,CONH),7.64(s,1H,CCHN),7.34(m,1H,H-3),7.30-7.27(m,1H,H -5),7.25(d,J=8.4Hz,1H,H-6),5.81(m,1H,C(CH)O),5.54(d,J=14.7Hz,2H,NCH2C),4.61(m,2H,NHC H2),2.21(d,J=16.4Hz,2H,COCH2N),2.48(s,3H,CH3),2.44(s,3H,CH3),2.43(s,3H,CH3),2.26(s,,6H,N(CH3)2,0.81(t,J=6.7Hz,3H,CH3); 13 C-NMR (150MHz, CDCl3) δ: 171.0, 167.8, 152.1, 149.6, 148.9, 144.7, 143.6, 137.1, 137.0, 133.9, 130. 5,127.8,127.7,122.5,74.0,60.4,53.1,45.2(2C),36.3,35.6,27.5,22.3,21.7,21.5,20.7,13.9.;

[0211] HRMS:Calcd.for C 27 H 36 ClN7O3(M+H):542.2641.Found::542.2641.

[0212] Example 16: Preparation of compound B16

[0213] The structural formulas of compound B16 and its levorotatory isomer, compound B16(S), are as follows:

[0214]

[0215] Referring to Example 1, only 3-iodo-butylphthalide was replaced with butylphthalide, and morpholine was replaced with dimethylamine. The product was a yellow viscous liquid with a yield of 21%. Compound B16 and its levorotatory isomer B16(S) were also discussed. 1 H NMR, 13 The C NMR, HRMS, and specific rotation values ​​are shown below.

[0216] -24.8 (MeOH) 1 H-NMR(600MHz, CDCl3)δ:7.65(s,1H,CCHN),7.60(m,1H,CONH),7.36-7.33(m,1H,H-3),7.33-7.32(m,1H,H-5),7. 22-7.31(m,1H,H-6),7.24-7.21(m,1H,H-4),5.88(m,1H,C(CH)O),5.53(d,J=14.6Hz,2H,NCH2C),4.62(m,2H,NHC H2),3.11(d,J=16.6Hz,2H,COCH2N),2.47(s,3H,CH3),2.43(s,3H,CH3),2.42(s,3H,CH3),2.23(s,6H,N(CH3)2,0.76(t,J=7.2Hz,3H,CH3); 13 C-NMR (150MHz, CDCl3) δ:169.8,168.2,151.0,148.5,147.9,144.0,142.6,137.5,134.4,129.3,127 .0,126.6,125.1,121.5,73.5,59.3,52.1,44.1(2C),35.4,34.5,26.6,21.3,20.7,20.5,19.6,12.9.

[0217] HRMS:Calcd.for C 27 H 37 ClN7O3(M+H):508.3031.Found::508.3031.

[0218] Example 17: Preparation of compound B17

[0219] The structural formulas of compound B17 and its levorotatory isomer, B17(S), are as follows:

[0220]

[0221] Accurately weigh 1 g (1.6 mmol) of intermediate 5 into a microwave tube, dissolve it in 3 mL of tetrahydrofuran, and then add cesium carbonate (0.9 g, 3.8 mmol) and 1-Boc-piperazine (0.5 g, 3.8 mmol) sequentially. React in a microwave-safe environment at 65 °C for 1 h. After the reaction is complete as detected by TLC, filter and concentrate to obtain the crude product. Purify the crude product by silica gel column chromatography (V:V:ethyl acetate = 8:1) to obtain a yellow viscous liquid B17-Boc. Add trifluoroacetic acid and react at room temperature for 1 h to obtain a yellow viscous liquid B17, with a yield of 48%. The compounds B17-Boc, B17, and the levorotatory isomer B17(S) are also discussed. 1 H NMR, 13 The C NMR, HRMS, and specific rotation values ​​are shown below.

[0222] B17-Boc: -50.8 (MeOH) 1H-NMR(600MHz,CDCl3)δ:7.74(d,J=1.7Hz,1H,H-6),7.71(dd,J=8.3Hz,1.9Hz,1H,H-5),7.71(s,1H,CCHN),7.65(t,J=5.8Hz,1H,CONH),7.10(d,J=8.3Hz,1H,H-3),5.86(m,1H,C(CH)O),5.61(d,J=14.6Hz,,2H,NCH2C),4.67(m,2H,NHC H 2),3.43(t,J=4.5Hz,4H,CH2NCH2),3.20(d,J=16.5Hz,2H,COCH2N),2.55(s,3H,CH3),2.51(s,3H,CH3),2.50(s,3H,CH3),2.49-2.40(m,4H,C H 2N(CO)C H 2),1.45(s,9H,Boc-(CH3)3),0.83(t,J=7.1Hz,3H,CH3); 13 C-NMR(150MHz,CDCl3)δ:170.6,167.4,154.6,152.1,149.6,148.9,144.6,143.5,139.4,138.3,137.2,136.4,127.8,122.6,93.4,79.8,74.2,59.3,53.1,52.7,36.3,35.5,28.4,27.4,22.3,21.8,21.6,20.7,13.9.;HRMS:Calcd.forC 34 H 47 IN8O5(M+H):775.2787.Found:775.2787.

[0223] B17: -31.7(MeOH) 1 H-NMR(600MHz,CDCl3)δ:7.68-7.66(m,1H,CONH),7.66-7.65(m,1H,H-6),7.65-7.64(m,1H,CCHN),7.64-7.63(m,1H,H-5),7.04(d,J=8.2Hz,1H,H-3),5.82(m,1H,C(CH)O),5.53(d,J=14.6Hz,2H,NCH2C),4.57(d,J=5.7Hz,2H,NHC H2),3.20(d,J=17.0Hz,2H,COCH2N),3.11(t,J=4.9Hz,4H,CH2NCH2),2.71(m,4H,C H 2N(CO)C H 2),2.47(s,3H,CH3),2.44(s,3H,CH3),2.43(s,3H,CH3),0.76(t,J=7.3Hz,3H,CH3); 13 C-NMR (150MHz, CDCl3) δ:168.9,166.4,151.1,148.6,147.8,143.5,142.4,138.4,137.3,136.0,135.2,126.8 ,121.8,92.3,73.2,57.4,52.1,48.1,42.3,35.1,34.3,26.4,21.2,20.7,20.5,19.6,12.8.; HRMS:Calcd.for C 29 H 39 IN8O3(M+H):675.2263.Found:675.2263.

[0224] Example 18: Preparation of compound B18

[0225] The structural formulas of compound B18 and its levorotatory isomer, B18(S), are as follows:

[0226]

[0227] Referring to Example 17, only 3-iodo-butylphthalide was replaced with 3-bromo-butylphthalide, and the product was a yellow viscous liquid with a yield of 35%. Compound B18-Boc and compound B18 and its levorotatory isomer B18(S) 1 H NMR, 13 The C NMR, HRMS, and specific rotation values ​​are shown below.

[0228] B18-Boc: -54.3 (MeOH) 1 H-NMR (600MHz, CDCl3) δ: 7.72 (s, 1H, CCHN), 7.68 (t, J = 5.8Hz, 1H, CONH), 7.55 (d, J = 2.0Hz, 1H, H-3), 7.52 (dd, J = 8.4 Hz,2.0Hz,1H,H-5),7.24(d,J=8.4Hz,1H,H-6),5.88(m,1H,C(CH)O),5.61(d,J=14.6Hz,2H,NCH2C),4.67(m,2H,NHC H2),3.43(t,J=5.0Hz,4H,CH2NCH2),3.21(d,J=16.4Hz,2H,COCH2N),2.55(s,3H,CH3),2.51(s,3H,CH3),2.50(s,3H,CH3),2.49-2.41(m,4H,C H 2N(CO)C H 2),1.45(s,9H,Boc-(CH3)3),0.83(t,J=7.1Hz,3H,CH3); 13 C-NMR(150MHz,CDCl3)δ:170.6,167.6,154.6,152.1,149.6,148.9,144.6,143.5,137.6,137.1,133.5,130.6,127.8,122.6,121.9,,79.8,74.1,59.3,53.1,52.7,36.3,35.5,28.4,27.4,22.3,21.7,21.5,20.7,13.9.;HRMS:Calcd.forC 34 H 47 BrN8O5(M+H):727.2925.Found:727.2925.

[0229] B18: -31.2(MeOH) 1 H-NMR(600MHz,CDCl3)δ:7.65(s,1H,CCHN),7.60(t,J=5.9Hz,1H,CONH),7.48(d,J=2.0Hz,1H,H-3),7.46(dd,J=8.5Hz,2.2Hz,1H,H-5),7.19(d,J=8.4Hz,1H,H-6),5.84(m,1H,C(CH)O),5.54(d,J=14.6Hz,2H,NCH2C),4.59(d,J=6.0Hz,2H,NHC H 2),3.21(d,J=16.9Hz,2H,COCH2N),3.13(t,J=5.1Hz,4H,,CH2NCH2),2.75(m,4H,C H 2NHC H 2),2.47(s,3H,CH3),2.44(s,3H,CH3),2.43(s,3H,CH3),0.76(t,J=7.1Hz,3H,CH3); 13C-NMR (150MHz, CDCl3) δ:168.9,166.5,151.1,148.6,147.8,143.5,142.4,136.6,136.0,132.5,129.5,126.8 ,121.7,120.9,73.2,57.5,52.1,48.1,42.4,35.1,34.4,26.4,21.2,20.7,20.5,19.7,12.9.; HRMS:Calcd.for C 29 H 39 BrN8O3(M+H):627.2402.Found:627.2402.

[0230] Example 19: Preparation of compound B19

[0231] The structural formulas of compound B19 and its levorotatory isomer, compound B19(S), are as follows:

[0232]

[0233] Referring to Example 17, only 3-iodo-butylphthalide was replaced with 3-chloro-butylphthalide; the product was a yellow viscous liquid with a yield of 20%. Compound B19-Boc and compound B19 and its levorotatory isotropic form B19(S) were also discussed. 1 H NMR, 13 The C NMR, HRMS, and specific rotation values ​​are shown below.

[0234] B19-Boc: -55.2 (MeOH) 1 H-NMR (600MHz, CDCl3) δ: 7.64 (s, 1H, CCHN), 7.61 (t, J = 5.8Hz, 1H, CONH), 7.33 (d, J = 2.1Hz, 1H, H-3), 7.29 (dd, J = 8.4 Hz,2.2Hz,1H,H-5),7.23(d,J=8.4Hz,1H,H-6),5.82(m,1H,C(CH)O),5.54(d,J=14.5Hz,2H,NCH2C),4.60(m,2H,NHC H 2),3.36(t,4H,CH2NCH2),3.13(d,J=16.7Hz,2H,COCH2N),2.48(s,3H,CH3),2.44(s,3H,CH3),2.43(s,3H,CH3),2.42-2.32(m,4H,C H 2N(CO)C H2),1.38(s,9H,Boc-(CH3)3),0.76(t,J=7.2Hz,3H,CH3); 13 C-NMR(150MHz,CDCl3)δ:170.7,167.7,154.6,,152.1,149.6,148.9,144.6,143.5,137.1,136.9,133.9,130.6,127.7,127.6,122.6,79.8,74.1,59.4,53.1,52.7,36.4,35.5,28.4,27.5,22.3,21.8,21.5,20.7,13.9.;HRMS:Calcd.for C 34 H 47 ClN8O5(M+H):683.3431.Found:683.3431.

[0235] B19: -33.8(MeOH) 1 H-NMR(600MHz,CDCl3)δ:7.70(t,J=5.6Hz,1H,CONH),7.66(s,1H,CCHN),7.32(d,J=2.2Hz,1H,H-3),7.30(dd,J=8.4Hz,2.2Hz,1H,H-5),7.24(d,J=8.4Hz,1H,H-6),5.85(m,1H,C(CH)O),5.53(d,J=14.6Hz,2H,NCH2C),4.58(d,J=5.6Hz,2H,NHC H 2),3.20(d,J=16.9Hz,2H,COCH2N),3.11(t,J=7.3Hz,4H,,CH2NCH2),2.71(m,4H,C H 2NHC H 2),2.46(s,3H,CH3),2.44(s,3H,CH3),2.42(s,3H,CH3),0.76(t,J=7.2Hz,3H,CH3); 13 C-NMR(150MHz,CDCl3)δ:169.9,167.7,152.2,149.7,148.8,144.6,143.5,137.1,136.8,133.9,130.6,127.7,127.6,122.8,74.1,58.5,53.1,49.1,43.4,36.2,35.4,27.5,22.3,21.7,21.5,20.7,13.9.

[0236] HRMS:Calcd.for C29 H 39 ClN8O3(M+H):583.2906.Found::583.2906.

[0237] Example 20: Preparation of compound B20

[0238] The structural formulas of compound B20 and its levorotatory isomer, B20(S), are as follows:

[0239]

[0240] Referring to Example 17, only 3-iodo-butylphthalide was replaced with butylphthalide; the product was a yellow viscous liquid with a yield of 60%. Compound B20-Boc and compound B20 and its levorotatory isotropic form B20(S) were also discussed. 1 H NMR, 13 The C NMR, HRMS, and specific rotation values ​​are shown below.

[0241] B20-Boc: -47.8 (MeOH) 1 H-NMR (600MHz, CDCl3) δ: 7.70 (s, 1H, CONH), 7.59 (t, J = 5.8Hz, 1H, CCHN), 7.41-7.39 (m, 1H, H-3), 7.39-7.36 (m, 1H, H-5 ),7.36-7.33(m,1H,H-6),7.30-7.26(m,1H,H-4),5.92(m,1H,C(CH)O),5.59(d,J=14.5Hz,2H,NCH2C),4.67(m,2H,NHC H 2),3.41(t,J=4.9Hz,4H,CH2NCH2),3.19(d,J=16.7Hz,2H,COCH2N),2.53(s,3H,CH3),2.49(s,3H,CH3),2.48(s,3H,CH3),2.47-2.37(m,4H,C H 2N(CO)C H 2),1.43(s,9H,Boc-(CH3)3),0.76(t,J=7.3Hz,3H,CH3); 13C-NMR(150MHz,CDCl3)δ:169.5,168.1,153.6,151.0,148.5,147.9,143.9,142.5,137.6,134.3,129.4,127.0,126.6,125.0,121.5,78.7,73.5,51.7,35.4,34.4,27.3,26.5,21.3,20.7,20.5,19.6,12.9.;HRMS:Calcd.for C 34 H 48 N8O5(M+H):649.3819.Found:649.3819.

[0242] B20: -27.8(MeOH) 1 H-NMR(600MHz,CDCl3)δ:7.66(s,1H,CONH),7.58(t,J=5.8Hz,1H,CCHN),7.36-7.34(m,1H,H-3),7.34-7.31(m,1H,H-5),7.31-7.28(m,1H,H-6),7.24-7.20(m,1H,H-4),5.89(m,1H,C(CH)O),5.54(d,J=14.5Hz,2H,NCH2C),4.60(d,J=5.8Hz,2H,NHC H 2),3.18(d,J=16.6Hz,2H,COCH2N),3.04(t,J=5.1Hz,4H,CH2NCH2),2.64(m,4H,C H 2N(CO)C H 2),2.47(s,3H,CH3),2.44(s,3H,CH3),2.43(s,3H,CH3),0.82(t,J=7.2Hz,3H,CH3); 13 C-NMR(150MHz,CDCl3)δ:169.0,168.1,151.1,148.6,147.9,143.8,142.5,137.6,134.2,129.4,127.0,126.6,125.0,121.6,73.6,57.9,52.1,49.3,42.9,35.3,34.4,28.7,26.6,21.3,20.7,20.5,19.6.

[0243] HRMS:Calcd.for C 29 H 40 N8O3(M+H):549.3296.Found:549.3296.

[0244] Pharmacological studies of triazole compounds used to protect brain tissue.

[0245] This invention utilizes the MTT assay to evaluate the protective effect of compounds on SH-SY5Y cells (neurons): A hypoxia-reoxygenation model of SY5Y cells was established to simulate cerebral ischemia-reperfusion injury. SH-SY5Y cells in logarithmic growth phase were seeded at 1.2 × 10⁴ cells per well in 96-well plates and cultured at 5% CO₂ and 37°C for approximately 24 hours until cell coverage reached 80%. Then, different treatment groups were selected as follows:

[0246] ① Blank control group: Replace with serum-free high-glucose culture medium and incubate in a normal cell culture incubator.

[0247] ② Hypoxia-reoxygenation model group (model group): The serum-free high-glucose medium was replaced and cultured in a normal cell culture incubator for 4 hours. Then, the medium was replaced with serum-free and glucose-free medium and placed in an anaerobic incubator for 3.5 hours to induce hypoxic injury. The cells were then removed from the anaerobic incubator, replaced with serum-free high-glucose medium, and cultured in a normal cell culture incubator for another 12 hours.

[0248] ③ Triazole compound group for protecting brain tissue (pretreatment group): The culture medium was replaced with serum-free high-glucose medium containing different concentrations of one triazole compound B1-20 for protecting brain tissue. After incubation in a normal cell culture incubator for 4 hours, the medium was replaced with serum-free and glucose-free medium and placed in an anaerobic incubator for 3.5 hours to induce hypoxic injury. Then, the culture was removed from the anaerobic incubator, replaced with serum-free high-glucose medium, and placed in a normal cell culture incubator for 12 hours.

[0249] After the above three treatments were completed, 10 μL of CCK-8 solution was added to each well, and the mixture was incubated at 37°C for 1 hour. The absorbance (OD value) was then measured at 450 nm using a microplate reader.

[0250] Cell protection rate % = (OD value of pretreatment group ③ - OD value of model group ②) / (OD value of blank control group ① - OD value of model group ②) × 100%; the data are shown in Table 1:

[0251] Table 1. Evaluation of the protective activity of compound B1-20 against the SY5Y cell injury model.

[0252]

[0253] The results of the protective activity experiments of the compounds against the SY5Y cell injury model showed that, compared with the model group, compounds B1, B2, B3, B4, B5, B6, B7, B8, B10, B12, B14, B16, B18, and B20 all exhibited protective activity against the SH-SY5Y cell injury model. Among them, compound B4 showed the most outstanding activity, with a nerve cell protection rate of 58.59% at a concentration of only 1.56 μM and a nerve cell protection rate as high as 100.23% at 12.5 μM. Compounds B8, B12, B16, and B20 also showed very significant protective activity against the SH-SY5Y cell injury model, with nerve cell protection rates all exceeding 44% at specific concentrations.

[0254] It is evident that the triazole compounds for protecting brain tissue provided by this invention have good development prospects and can be used to effectively protect brain tissue and prevent and treat ischemic stroke.

[0255] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, and is not intended to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions made by those skilled in the art to the technical solution of the present invention do not depart from the essence and scope of the technical solution of the present invention.

Claims

1. A triazole compound for protecting brain tissue, characterized in that, The structural formula is any one of the following: 、 、 、 、 、 、 、 、 、 、 、 、 、 。 2. The method for preparing the triazole compound according to claim 1, characterized in that, Includes the following steps: (1) Butylphthalide or halogen-substituted butylphthalide is hydrolyzed with alkaline solution, pH is adjusted, concentration is achieved, and chloroacetyl chloride, 4-dimethylaminopyridine and triethylamine are added to react and intermediate 1 is obtained. (2) Dissolve intermediate 1 obtained in step (1) with propargylamine, add O-benzotriazole-N,N,N',N'-tetramethylurea tetrafluoroboric acid and N,N-diisopropylethylamine to react and obtain intermediate 2; (3) Ligustrazine was brominated with NBS using benzoyl peroxide as an initiator to obtain intermediate 3; (4) Dissolve intermediate 3 obtained in step (3), and then add sodium azide to react and obtain intermediate 4; (5) Dissolve intermediate 2 obtained in step (2) and intermediate 4 obtained in step (4), add catalyst to react and obtain intermediate 5; (6) The intermediate 5 obtained in step (5) undergoes a substitution reaction with the substituted compound under the action of a catalyst to obtain the triazole compound; The intermediate 1 is The intermediate 2 is ; The intermediate 3 is The intermediate 4 is The intermediate 5 is Where X is H, I, or Br.

3. The preparation method according to claim 2, characterized in that, The alkali used in step (1) is selected from at least one of sodium hydroxide, potassium hydroxide, and calcium hydroxide; the hydrolysis conditions in step (1) are: incubation at 55-65℃ for 1.5-3 hours, or microwave treatment at 55-65℃ for 30-45 minutes; the target pH for pH adjustment in step (1) is 3-4, and the reagent used is selected from one or more of dilute hydrochloric acid, dilute sulfuric acid, and dilute nitric acid; the concentration method in step (1) is extraction by adding an extractant; the extractant is selected from ethyl acetate and diethyl ether. Or two; the molar ratio of chloroacetyl chloride, 4-dimethylaminopyridine and triethylamine to butylphthalide or halogen-substituted butylphthalide in step (1) is 1.4-1.6:0.08-0.12:1.4-1.6:1; the molar ratio of propargylamine to intermediate 1 in step (2) is 1.1-1.3:1; the molar ratio of O-benzotriazole-N,N,N',N'-tetramethylurea tetrafluoroboric acid, N,N-diisopropylethylamine to intermediate 1 in step (2) is: 1.1-1.3：1.8-2.2：1。 4. The preparation method according to claim 2, characterized in that, The molar ratio of benzoyl peroxide to tetramethylpyrazine in step (3) is 0.08-0.12:1; the molar ratio of NBS to tetramethylpyrazine in step (3) is 0.25-0.35:1; the molar ratio of sodium azide to intermediate 3 in step (4) is 1.15-1.25:1; the reaction conditions in step (4) are microwave at 55-65℃ for 55-70 min.

5. The preparation method according to claim 2, characterized in that, The molar ratio of intermediate 2 to intermediate 4 in step (5) is 1:0.9-1.1; the catalyst in step (5) includes cuprous thiophene-2-carboxylate (I); the molar ratio of the substituted compound to intermediate 5 in step (6) is 1.4-1.6:1; the substituted compound in step (6) is selected from morpholine, N-methylpiperazine, piperazine, diethylamine and dimethylamine; the substitution reaction conditions in step (6) are 60-70℃ for 100-140 min; or microwave at 60-70℃ for 50-65 min.

6. A pharmaceutical composition comprising a triazole compound or a pharmaceutically acceptable salt as described in claim 1.

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