Tetrazole derivatives, preparation, pharmaceutical compositions containing them and their use

Abstract

The application discloses a tetrazole derivative, which has a structure shown in a general formula I. The application also discloses a composition containing the tetrazole derivative and application of the tetrazole derivative in preparation of a drug for preventing and resisting stroke. The inventors of the application have confirmed through multiple experiments that the compound has obvious inhibiting effect on ADP-induced platelet aggregation activity, outstanding damage protection effect on OGD / R fetal rat primary cortical neuron cells and neuron-like cells induced and differentiated from neuroblastoma cells (N2A), good anti-stroke efficacy on an animal model of focal cerebral ischemia constructed by a line plug method for temporarily blocking a middle cerebral artery and good rat oral pharmacokinetic properties. Therefore, the compound can be applied to a drug for treating and preventing stroke as a neuroprotective agent.

Description

Technical Field

[0001] This invention relates to the field of pharmaceuticals, specifically to tetrazolium derivatives, preparation methods, pharmaceutical compositions containing them, and their use in the preparation of drugs for the prevention and treatment of cardiovascular and cerebrovascular diseases, the improvement of cardiovascular and cerebrovascular circulation disorders, or the prevention of thrombosis. Background Technology

[0002] Stroke is one of the most common cardiovascular and cerebrovascular diseases, and has become the leading cause of disability and the second leading cause of death among adults worldwide. Ischemic stroke accounts for approximately 80% of all strokes, seriously threatening human health. In the past decade, research has primarily focused on exploring more effective treatment strategies to reduce stroke-related deaths and disabilities. Various drugs have been explored in clinical trials and animal models, but effective treatment strategies remain lacking. Furthermore, there are currently no neuroprotective agents for the prevention of acute ischemic stroke, thus requiring novel compounds with new structural features and mechanisms of action. This field has a broad market and clinical demand. Tetraazole derivatives have wide applications in drug development, demonstrating good efficacy in antihypertensive, anti-inflammatory, antibacterial, platelet aggregation inhibitor, and asthma treatment. For example, the antihypertensive drug Losartan, the first orally active and potent non-peptide angiotensin II receptor antagonist, and the platelet aggregation inhibitor Cilostazol, both incorporate tetraazole structures and have achieved very good pharmacological effects. Summary of the Invention

[0003] The purpose of this invention is to provide a tetrazolium derivative and its pharmaceutically acceptable salt or solvate.

[0004] The present invention also provides a method for preparing the above-mentioned tetrazolium derivatives.

[0005] The present invention also provides a pharmaceutical composition containing the above-mentioned tetrazolium derivatives and their pharmaceutically acceptable salts or solvates.

[0006] The present invention also provides an application of the above-mentioned tetrazolium derivatives and their pharmaceutically acceptable salts or solvates in the preparation of drugs for the prevention and treatment of stroke.

[0007] The present invention adopts the following technical solution:

[0008] This invention provides a tetrazolium derivative, characterized in that it has the structure shown in general formula (I):

[0009]

[0010] Or its optical isomers;

[0011] Or its pharmaceutically acceptable salts or solvates;

[0012] Wherein: R1 is H, a C1-C3 alkyl chain or a haloalkyl chain;

[0013] R2 is H, amino, nitro, hydroxyl, ether, methyl, ester, carbonyl, -CF3, -OCF3, n-butyl, isopropyl, peptide, one or more Cl, one or more F, one or more Br, or a combination formed by at least two of one or more Cl, one or more F, and one or more Br;

[0014] R3 is selected independently.

[0015] Where R4 is H or -C(=O)C1-C3 alkyl;

[0016] A is a benzene ring, a six-membered heterocycle containing at least one nitrogen atom, or a five-membered heterocycle containing at least one nitrogen atom.

[0017] Preferably, when R1 is H and A is a benzene ring, R2 is not H, F, Cl, or Br;

[0018] Furthermore, as a preferred embodiment, the tetrazolium derivatives of the present invention have the structure shown in general formula (II):

[0019]

[0020] Or its optical isomer; or its pharmaceutically acceptable salt or solvate;

[0021] R1, R2, and R3 are defined as above; X is C or N.

[0022] Furthermore, preferably, the tetrazolium derivative has the structure shown in general formula III:

[0023]

[0024] Or its optical isomer; or its pharmaceutically acceptable salt or solvate; or its optical isomer and pharmaceutically acceptable salt or solvate;

[0025] Preferably, R1 is H, methyl, or ethyl; R2 is H, Cl, F, Br, or amino.

[0026] X is C or N.

[0027] Preferably, R1 is methyl or ethyl; preferably, X is N.

[0028] Further, as a preferred embodiment, the pharmaceutically acceptable salt is preferably a salt composed of one or more of sodium hydroxide, potassium hydroxide, lithium hydroxide, sodium methoxide, potassium methoxide, sodium ethoxide, potassium ethoxide, triethylamine, and tert-butylamine.

[0029] More preferably, sodium hydroxide and potassium hydroxide have the general formula Va and Vb structures:

[0030]

[0031] Or its optical isomers, wherein X and R2 are defined as above.

[0032] Preferably, the tetrazolium derivatives include one or more of the following compounds:

[0033] 1-(4-bromo-2-(1-methyl-1H-tetrazol-5-yl)phenyl)pentan-1-ol;

[0034] 1-(6-bromo-2-(1H-tetrazol-5-yl)pyridin-3-yl)pentan-1-ol;

[0035] 1-(6-bromo-2-(1-methyl-1H-tetrazol-5-yl)pyridin-3-yl)pentan-1-ol;

[0036] 1-(4-chloro-2-(1-methyl-1H-tetrazol-5-yl)phenyl)pentan-1-ol;

[0037] 1-(6-chloro-2-(1H-tetrazol-5-yl)pyridin-3-yl)pentan-1-ol;

[0038] 1-(6-chloro-2-(1-methyl-1H-tetrazol-5-yl)pyridin-3-yl)pentan-1-ol;

[0039] 1-(4-fluoro-2-(1-methyl-1H-tetrazol-5-yl)phenyl)pentan-1-ol;

[0040] 1-(6-fluoro-2-(1H-tetrazol-5-yl)pyridin-3-yl)pentan-1-ol;

[0041] 1-(6-fluoro-2-(1-methyl-1H-tetrazol-5-yl)pyridin-3-yl)pentan-1-ol;

[0042] 1-(2-(1-methyl-1H-tetrazol-5-yl)phenyl)pentan-1-ol;

[0043] 1-(2-(1H-tetrazol-5-yl)pyridin-3-yl)pentan-1-ol;

[0044] 1-(2-(1-methyl-1H-tetrazol-5-yl)pyridin-3-yl)pent-1-ol;

[0045] 1-(4-amino-2-(1H-tetrazol-5-yl)phenyl)pentan-1-ol;

[0046] 1-(4-amino-2-(1-methyl-1H-tetrazol-5-yl)phenyl)pentan-1-ol;

[0047] 1-(4-nitro-2-(1H-tetrazol-5-yl)phenyl)pentan-1-ol;

[0048] 1-(4-nitro-2-(1-methyl-1H-tetrazol-5-yl)phenyl)pentan-1-ol;

[0049] 1-(2-(1H-tetrazol-5-yl)-4-(trifluoromethyl)phenyl)pentan-1-ol;

[0050] 1-(2-(1-methyl-1H-tetrazol-5-yl)-4-(trifluoromethyl)phenyl)pentan-1-ol;

[0051] 1-(4-bromo-2-(1H-tetrazol-5-yl)phenyl)pentan-1-ol;

[0052] 1-(4-bromo-2-(1H-tetrazol-5-yl)phenyl)but-1-ol;

[0053] 1-(4-bromo-2-(1-methyl-1H-tetrazol-5-yl)phenyl)but-1-ol;

[0054] 1-(4-bromo-2-(1H-tetrazol-5-yl)phenyl)hex-1-ol;

[0055] 1-(4-bromo-2-(1-methyl-1H-tetrazol-5-yl)phenyl)hex-1-ol;

[0056] 1-(4-chloro-2-(1H-tetrazol-5-yl)phenyl)pentan-1-ol;

[0057] 1-(4-chloro-2-(1H-tetrazol-5-yl)phenyl)but-1-ol;

[0058] 1-(4-chloro-2-(1-methyl-1H-tetrazol-5-yl)phenyl)but-1-ol;

[0059] 1-(4-chloro-2-(1H-tetrazol-5-yl)phenyl)hex-1-ol;

[0060] 1-(4-chloro-2-(1-methyl-1H-tetrazol-5-yl)phenyl)hex-1-ol;

[0061] 1-(4-fluoro-2-(1H-tetrazol-5-yl)phenyl)pentan-1-ol;

[0062] 1-(4-fluoro-2-(1H-tetrazol-5-yl)phenyl)but-1-ol;

[0063] 1-(4-fluoro-2-(1-methyl-1H-tetrazol-5-yl)phenyl)but-1-ol;

[0064] 1-(4-fluoro-2-(1H-tetrazol-5-yl)phenyl)hex-1-ol;

[0065] 1-(4-fluoro-2-(1-methyl-1H-tetrazol-5-yl)phenyl)hex-1-ol;

[0066] 1-(2-(1H-tetrazol-5-yl)phenyl)pentan-1-ol;

[0067] 1-(2-(1H-tetrazol-5-yl)phenyl)but-1-ol;

[0068] 1-(2-(1-methyl-1H-tetrazol-5-yl)phenyl)but-1-ol;

[0069] 1-(2-(1H-tetrazol-5-yl)phenyl)hex-1-ol;

[0070] 1-(2-(1-methyl-1H-tetrazol-5-yl)phenyl)hex-1-ol;

[0071] 1-(5-bromo-2-(1H-tetrazol-5-yl)phenyl)pentan-1-ol;

[0072] 1-(5-bromo-2-(1H-tetrazol-5-yl)phenyl)hex-1-ol

[0073] Or other optical isomers of the above compounds; or pharmaceutically acceptable salts or solvates of the above compounds; or pharmaceutically acceptable salts or solvates of their optical isomers.

[0074] Specifically, according to general formula I, the preferred compound of the present invention is:

[0075]

[0076]

[0077]

[0078]

[0079]

[0080]

[0081] And optical isomers of the above compounds; or pharmaceutically acceptable salts or solvates thereof.

[0082] Or other optical isomers of the above compounds; or pharmaceutically acceptable solvates of the above compounds.

[0083] Preferably, the pharmaceutically acceptable salt is a salt formed with one or more of the following bases: sodium hydroxide, potassium hydroxide, lithium hydroxide, sodium methoxide, potassium methoxide, sodium ethoxide, potassium ethoxide, triethylamine, and tert-butylamine.

[0084] Preferably, the tetrazolium derivatives include:

[0085] 1-(6-bromo-2-(1H-tetrazol-5-yl)pyridin-3-yl)pentan-1-ol sodium salt

[0086] 1-(6-bromo-2-(1H-tetrazol-5-yl)pyridin-3-yl)pent-1-ol potassium salt

[0087] 1-(6-bromo-2-(1H-tetrazol-5-yl)pyridin-3-yl)pentan-1-ol lithium salt

[0088] 1-(6-bromo-2-(1H-tetrazol-5-yl)pyridin-3-yl)pent-1-ol triethylamine salt

[0089] 1-(6-bromo-2-(1H-tetrazol-5-yl)pyridin-3-yl)pent-1-ol tert-butylamine salt

[0090] 1-(2-(1H-tetrazol-5-yl)-4-(trifluoromethyl)phenyl)pentan-1-ol sodium salt

[0091] 1-(2-(1H-tetrazol-5-yl)-4-(trifluoromethyl)phenyl)pentan-1-ol potassium salt

[0092] 1-(2-(1H-tetrazol-5-yl)-4-(trifluoromethyl)phenyl)pentane-1-ol lithium salt

[0093] 1-(2-(1H-tetrazol-5-yl)-4-(trifluoromethyl)phenyl)pent-1-ol triethylamine salt

[0094] 1-(2-(1H-tetrazol-5-yl)-4-(trifluoromethyl)phenyl)pent-1-ol tert-butylamine salt

[0095] 1-(4-amino-2-(1H-tetrazol-5-yl)phenyl)pentan-1-ol sodium salt

[0096] 1-(4-amino-2-(1H-tetrazol-5-yl)phenyl)pentan-1-ol potassium salt

[0097] 1-(4-amino-2-(1H-tetrazol-5-yl)phenyl)pentane-1-ol lithium salt

[0098] 1-(4-amino-2-(1H-tetrazol-5-yl)phenyl)pent-1-ol triethylamine salt

[0099] 1-(4-amino-2-(1H-tetrazol-5-yl)phenyl)pentane-1-ol tert-butylamine salt

[0100] 1-(4-bromo-2-(1H-tetrazol-5-yl)phenyl)pentan-1-ol sodium salt

[0101] 1-(4-bromo-2-(1H-tetrazol-5-yl)phenyl)pentan-1-ol potassium salt

[0102] 1-(4-bromo-2-(1H-tetrazol-5-yl)phenyl)pentane-1-ol lithium salt

[0103] 1-(4-bromo-2-(1H-tetrazol-5-yl)phenyl)pentane-1-ol triethylamine salt

[0104] 1-(4-bromo-2-(1H-tetrazol-5-yl)phenyl)pentane-1-ol tert-butylamine salt

[0105] Or other optical isomers of the above compounds; or pharmaceutically acceptable solvates of the above compounds.

[0106] Preferably, the tetrazolium derivatives include:

[0107] (R)-1-(4-bromo-2-(1H-tetrazol-5-yl)phenyl)pentan-1-ol

[0108] (S)-1-(4-bromo-2-(1H-tetrazol-5-yl)phenyl)pentan-1-ol

[0109] (R)-1-(4-chloro-2-(1H-tetrazol-5-yl)phenyl)pentan-1-ol

[0110] (S)-1-(4-chloro-2-(1H-tetrazol-5-yl)phenyl)pentan-1-ol

[0111] (R)-1-(4-fluoro-2-(1H-tetrazol-5-yl)phenyl)pentan-1-ol

[0112] (S)-1-(4-fluoro-2-(1H-tetrazol-5-yl)phenyl)pentan-1-ol

[0113] (R)-1-(4-bromo-2-(1H-tetrazol-5-yl)phenyl)hex-1-ol

[0114] (S)-1-(4-bromo-2-(1H-tetrazol-5-yl)phenyl)hex-1-ol

[0115] (R)-1-(4-chloro-2-(1H-tetrazol-5-yl)phenyl)hex-1-ol

[0116] (S)-1-(4-chloro-2-(1H-tetrazol-5-yl)phenyl)hex-1-ol

[0117] (R)-1-(4-fluoro-2-(1H-tetrazol-5-yl)phenyl)hex-1-ol

[0118] (S)-1-(4-fluoro-2-(1H-tetrazol-5-yl)phenyl)hex-1-ol

[0119] (R)-1-(5-bromo-2-(1H-tetrazol-5-yl)phenyl)pentan-1-ol

[0120] (S)-1-(5-bromo-2-(1H-tetrazol-5-yl)phenyl)pentan-1-ol

[0121] (R)-1-(5-bromo-2-(1H-tetrazol-5-yl)phenyl)hex-1-ol

[0122] (S)-1-(5-bromo-2-(1H-tetrazol-5-yl)phenyl)hex-1-ol

[0123] Or other optical isomers of the above compounds; or pharmaceutically acceptable solvates of the above compounds.

[0124] A method for preparing the aforementioned tetrazolium derivatives involves reacting compound (1) with an acyl protecting agent to obtain compound (2), then reacting compound (2) with sodium azide, followed by the addition of hydrochloric acid to simultaneously remove the protecting group and eliminate residual sodium azide, resulting in compound (3). Compound (3) is then reacted with a Grignard reagent to obtain the compound shown in formula (I). Alternatively, the compound shown in formula (I) may be further reacted with an alkylating agent to obtain a tetrazolium alkyl-substituted product as shown in formula (I').

[0125]

[0126] Preferably, the acyl protecting agent is ethylene glycol.

[0127] The compound (1) can be obtained by bromination and hydrolysis of the compound of formula (1-1) as a raw material:

[0128]

[0129] When preparing compound (1-2) from compound (1-1), NBS is used as the brominating agent, and AIBN can be added as an initiator. Carbon tetrachloride is used as the reaction solvent, the reaction temperature is reflux temperature, and the reaction time is 10-24 hours.

[0130] The molar ratio of the brominating agent to compound (1-1) was 1.5–3:1. After the reaction was complete, water and ethanol (1:1) and an equivalent amount of silver nitrate as the compound were added directly to carry out the second hydrolysis reaction at a temperature of 60–80 °C for 4 hours.

[0131] Preferably, the reaction temperature of compound (1) with the acyl protecting agent is 70–130°C; the reaction agent can be toluene, and the catalyst can be p-toluenesulfonic acid. The molar ratio of ethylene glycol to compound (1) is 1–10:1, more preferably 2–8:1, and even more preferably 4–7:1. The molar ratio of p-toluenesulfonic acid to compound (1) is 0.03–0.2:1, more preferably 0.05–0.1:1.

[0132] Preferably, the reaction temperature of compound (2) with sodium azide is 110–170°C. Ammonium chloride is added, DMF is used as the reaction solvent, and the reaction is quenched with dilute hydrochloric acid after completion. The molar ratio of sodium azide to compound (2) is 2–10:1, more preferably 4–7:1. The molar ratio of ammonium chloride to compound (2) is 2–10:1, more preferably 4–7:1.

[0133] The reagent can be a commercially available product or prepared on-site. It can be obtained by reacting brominated C3-C5 alkanes with magnesium, and iodine particles can be added during the preparation process. The molar ratio of the brominated C3-C5 alkanes to compound (2) is calculated to be 2 to 10:1, and more preferably 4 to 7:1.

[0134] Preferably, the reaction temperature of compound (3) with the alkylating agent (iodotin) is 30–50 °C, the reaction solvent is DMF, and the catalyst is sodium hydroxide. The molar ratio of the iodoalkane to compound (3) is 1–2:1, more preferably 1–1.5:1. The molar ratio of sodium hydroxide to compound (3) is 2–3:1, more preferably 2.5–3:1.

[0135] A pharmaceutical composition comprising at least one active ingredient and one or more pharmaceutically acceptable carriers or excipients, wherein the active ingredient is a tetrazolium derivative as described in any of the above-mentioned technical solutions.

[0136] The use of any of the above-mentioned tetrazolium derivatives in the preparation of drugs for preventing and combating cardiovascular and cerebrovascular diseases, improving cardiovascular and cerebrovascular circulation disorders, or preventing thrombosis.

[0137] The term “pharmaceutically acceptable derivative” as used in this article refers to the salts and solvates of the selected compounds.

[0138] As used herein, the term "alkyl" refers to a straight-chain or branched alkane group containing carbon atoms. Examples of "alkyl" as used herein include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, etc. "alkyl" also includes substituted alkyl groups. Alkyl groups may optionally be substituted with halogens once or multiple times.

[0139] The term "halogen" as used herein refers to fluorine, chlorine, bromine, or iodine, preferably fluorine, chlorine, or bromine.

[0140] As used herein, the term "solvate" refers to a complex with variable stoichiometry formed by a solute (e.g., formula (I) of the present invention) and a solvent. For the purposes of the present invention, the solvent must not interfere with the biological activity of the solute. Examples of suitable solvents include, but are not limited to, water, methanol, ethanol, and acetic acid. Preferably, the solvent used is a pharmaceutically acceptable solvent. Suitable pharmaceutically acceptable solvents include, but are not limited to, water, ethanol, and acetic acid. More preferably, the solvent used is water.

[0141] The present invention can prepare the salts of the tetrazolium compounds described herein using methods well known to those skilled in the art. The salts can be organic or inorganic base salts, including sodium methoxide, potassium methoxide, sodium ethoxide, potassium ethoxide, triethylamine, tert-butylamine, etc.; and inorganic base salts including sodium hydroxide, potassium hydroxide, lithium hydroxide, etc.

[0142] A second object of the present invention is to provide a pharmaceutical composition comprising at least one active component and one or more pharmaceutically acceptable carriers or excipients, wherein the active component may be any one or more of the following: a tetrazolium compound with the structure shown in general formula (I) of the present invention and its preferred compounds, an optical isomer of the compound, a pharmaceutically acceptable salt of the compound or its optical isomer, or a solvate of the compound or its optical isomer.

[0143] The carrier includes one or more of the following conventional pharmaceutical diluents, excipients, fillers, binders, humectants, disintegrants, absorption enhancers, surfactants, adsorbents, lubricants, etc., and flavoring agents, sweeteners, etc. may be added if necessary. The drug of this invention can be formulated into various forms such as tablets, powders, granules, capsules, oral liquids, and injectable drugs, and all of the above dosage forms can be prepared according to conventional pharmaceutical methods.

[0144] The present invention also provides the use of the compound described in (I), its optical isomers or pharmaceutically acceptable salts or solvates thereof in the preparation of medicaments for the prevention and treatment of cardiovascular and cerebrovascular diseases, for improving cardiovascular and cerebrovascular circulatory disorders, or for antithrombotic purposes.

[0145] This invention also provides the use of the compounds described herein or their pharmaceutically acceptable salts in the preparation of remedies for the prevention and treatment of cardiovascular and cerebrovascular diseases, for improving cardiovascular and cerebrovascular circulatory disorders, or for antithrombotic purposes, particularly in the preparation of remedies for the treatment of acute ischemic stroke. In other words, this invention provides the use of tetrazolium compounds or their pharmaceutically acceptable salts, alone or in combination with other drugs, in the treatment of acute ischemic stroke. Anti-stroke remedies that can be used in combination with the compounds or their pharmaceutically acceptable salts provided by this invention include, but are not limited to, at least one of the following: free radical scavengers (such as edaravone); neuroprotective agents (such as septophan); antiplatelet drugs (such as clopidogrel, aspirin); and antithrombotic drugs (such as rivaroxaban).

[0146] Through multiple experiments, the inventors of this invention have confirmed that the compounds of this invention exhibit inhibitory activity against ADP-induced platelet aggregation and good oral pharmacokinetic properties in rats. Therefore, the compounds of this invention can be used as neuroprotective agents in the preparation of drugs for the treatment and prevention of acute ischemic stroke. Attached Figure Description

[0147] Figure 1 The structures of the positive control drugs butylphthalide (NBP) and BZP;

[0148] Figure 2 The protective effect of the compound on primary cortical neurons in fetal rats induced by OGD / R modeling;

[0149] Figure 3 The protective effect of the compound against neuron-like cells induced by OGD / R-induced neuroblastoma cells (N2A); where *p<0.05, ***p<0.001 vs. indicated group n=12. Error bars, SEM;

[0150] Figure 4 The pharmacodynamic effects of the compound in constructing a focal cerebral ischemia animal model by transient middle cerebral artery occlusion using online embolization; where #p<0.05, ##p<0.01, ###p<0.001 versus vehicle group; *p<0.05 versus NBP group. Error bars, SEM. Detailed Implementation

[0151] The feasibility of the present invention will be illustrated below through examples. Those skilled in the art should understand that modifications or substitutions to the corresponding technical features based on the teachings of the prior art still fall within the scope of protection claimed by the present invention.

[0152] Example 1. Synthesis of intermediates 1-3

[0153]

[0154] In a three-necked flask, starting material 1-1 (13.2 g, 0.1 mol), NBS (N-bromosuccinimide) (44.5 g, 0.25 mol), AIBN (azobisisobutyronitrile) (720 mg, 3 mmol), and carbon tetrachloride (20 mL) were added. The mixture was heated under reflux for 16 h, cooled to room temperature, and concentrated under reduced pressure to remove carbon tetrachloride, yielding a mixture containing intermediate 1-2. Water and ethanol (100 mL: 100 mL) were added to the mixture, and the reaction was carried out at 70 °C for 4 h. The organic phase was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to evaporate the ethyl acetate. The crude product was purified by silica gel column chromatography (ethyl acetate: petroleum ether = 1:50-1:10) to give approximately 8.4 g of white to pale yellow crystalline intermediate 1-3, with a yield of 40% and a purity greater than 99%. HRMS: m / z (ESI)calcd for C8H6N2O[M+H] + 147.05 found: 147.67.

[0155] Example 2: Synthesis of intermediates 1-5

[0156]

[0157] Step 1: In a single-necked flask, add intermediate 1-3 (2.8 g, 20 mmol), ethylene glycol (6.2 g, 0.1 mol), toluene (20 ml), and p-toluenesulfonic acid (275 mg, 1.6 mmol). Heat to reflux, removing water using a separator until the water level in the separator no longer increases. Cool to room temperature. Add saturated sodium bicarbonate solution until the aqueous layer is neutral or alkaline. Separate the aqueous layer, and concentrate the organic layer under reduced pressure to obtain 4.5 g of a pale yellow, oily, semi-solid intermediate 1-4. Yield: 89%. Purity: >99%.

[0158] Step 2: Intermediate 1-4 (0.7 g, 3.93 mmol), sodium azide (1.28 g, 19.65 mmol), ammonium chloride (1.05 g, 19.65 mmol), and DMF (10 mL) were added to a single-necked flask. The mixture was heated to reflux for 12 h, cooled to room temperature, quenched with 1 N hydrochloric acid, and stirred at room temperature for 1 h. After TLC confirmation of deprotection, the organic phase was extracted with ethyl acetate. The ethyl acetate was removed by concentration under reduced pressure. The crude product was purified by silica gel column chromatography (dichloromethane:methanol = 10:1) to obtain 924 mg of intermediate 1-5. Yield: 93%. HRMS: m / z (ESI) calcd for C8H7N5O[MH] - 189.07 found: 189.56.

[0159] Example 3: Synthesis of intermediates 2-3

[0160]

[0161] Following the steps of Example 1, 5-(trifluoromethyl)-2-methylbenzonitrile 2-1 (18.5 g, 0.1 mol) was used as the starting material to yield 15.2 g of solid 2-3, with a yield of 76% and a purity greater than 99%. HRMS: m / z (ESI) calcd for C9H4F3NO[M+H] + 200.02 found: 200.25.

[0162] Example 4: Synthesis of intermediates 2-5

[0163]

[0164] Referring to steps one and two of Example 2, intermediate 1-3 was replaced with intermediate 2-3, and intermediate 1-4 was replaced with intermediate 2-4, yielding 2.3 g of solid 2-5, with a yield of 56% (two steps), a purity greater than 99%, and HRMS: m / z (ESI) calcd for C9H5F3N4O[MH]. - 241.04 found: 240.89.

[0165] Example 5: Synthesis of Intermediate 3-1

[0166]

[0167] Following the steps of Example 1, 5-nitro-2-methylbenzonitrile 3-1 (16.2 g, 0.1 mol) was used as the starting material to obtain 8.9 g of solid 3-3, with a yield of 50% and a purity greater than 99%. HRMS: m / z (ESI) calcd for C8H4N2O3[M+H] + 177.02 found: 177.23.

[0168] Example 6: Synthesis of intermediates 3-5

[0169]

[0170] Referring to steps one and two of Example 2, intermediate 1-3 was replaced with intermediate 3-3, and intermediate 1-4 was replaced with intermediate 3-4, yielding 3.5 g of solid 3-5, with a yield of 72% (two steps), a purity greater than 99%, and HRMS: m / z (ESI) calcd for C8H5N5O3[MH]. - 218.04 found: 217.92.

[0171] Example 7: Synthesis of intermediate 4-3

[0172]

[0173] Following the steps of Example 1, using 19.5 g (0.1 mol) of 6-bromo-3-methylpyridinine 4-1 as a starting material, 6.2 g of solid 4-3 was obtained, with a yield of 29% and a purity greater than 99%. HRMS: m / z (ESI) calcd for C7H3BrN2O[M+H] + 210.94 found: 211.05.

[0174] Example 8: Synthesis of intermediates 4-5

[0175]

[0176] Referring to steps one and two of Example 2, intermediate 1-3 was replaced with intermediate 4-3, and intermediate 1-4 was replaced with intermediate 4-4, yielding 2.2 g of solid 4-5, with a yield of 35% (two steps), a purity greater than 99%, and HRMS: m / z (ESI) calcd for C7H4BrN5O[MH]. - 251.96 found: 252.05.

[0177] Example 9: Synthesis of Intermediate 5-3

[0178]

[0179] Following the steps of Example 1, using 19.4 g (0.1 mol) of 5-bromo-2-methylbenzonitrile 5-1 as a starting material, 6.2 g of solid 5-3 was obtained, with a yield of 29% and a purity greater than 99%. HRMS: m / z (ESI) calcd for C8H4BrNO[M+H] + 209.95 found: 210.10.

[0180] Example 10: Synthesis of Intermediate 5-5

[0181]

[0182] Referring to steps one and two of Example 2, intermediate 1-3 was replaced with intermediate 5-3, and intermediate 1-4 was replaced with intermediate 5-4, yielding 2.6 g of solid 5-5, with a yield of 37% (two steps), a purity greater than 99%, and HRMS: m / z (ESI) calcd for C8H5BrN4O[MH]. - 250.96 found: 251.32.

[0183] Example 11: Synthesis of intermediate 6-3

[0184]

[0185] Following the steps of Example 1, 5-chloro-2-methylbenzonitrile 6-1 (15.1 g, 0.1 mol) was used as the starting material to obtain 7.3 g of solid 6-3, with a yield of 44% and a purity greater than 99%. HRMS: m / z (ESI) calcd for C8H4ClNO[M+H] + 166.00 found: 166.23.

[0186] Example 12: Synthesis of intermediate 6-5

[0187]

[0188] Referring to steps one and two of Example 2, intermediate 1-3 was replaced with intermediate 6-3, and intermediate 1-4 was replaced with intermediate 6-4, yielding 3.2 g of white solid 6-5, with a yield of 65% (two steps) and a purity greater than 99%. HRMS: m / z (ESI) calcd for C8H5ClN4O[MH] - 207.02 found: 206.95.

[0189] Example 13: Synthesis of intermediate 7-3

[0190]

[0191] Following the steps of Example 1, 5-fluoro-2-methylbenzonitrile 7-1 (13.5 g, 0.1 mol) was used as the starting material to obtain 4.5 g of solid 7-3, with a yield of 30% and a purity greater than 99%. HRMS: m / z (ESI) calcd for C8H4FNO[M+H] + 150.03 found: 149.68.

[0192] Example 14: Synthesis of intermediate 7-5

[0193]

[0194] Referring to steps one and two of Example 2, intermediate 1-3 was replaced with intermediate 7-3, and intermediate 1-4 was replaced with intermediate 7-4, yielding 1.9 g of solid 7-5, with a yield of 39% (two steps), a purity greater than 99%, and HRMS: m / z (ESI) calcd for C8H5FN4O[MH]. - 191.04 found: 191.25.

[0195] Example 15: Synthesis of Intermediate 8-5

[0196]

[0197] Referring to steps one and two of Example 2, using 2-formylbenzonitrile 8-3 (13g, 0.1mol) as the starting material, intermediate 1-4 was replaced with intermediate 8-4 to obtain 6g of solid 8-5, with a yield of 34% (two steps) and a purity greater than 99%. HRMS: m / z (ESI) calcd for C7H5N5O[MH] - 173.05 found: 173.55.

[0198] Example 16: Synthesis of Intermediate 9-3

[0199]

[0200] Following the steps of Example 1, using (15.2 g, 0.1 mol) 6-chloro-3-methylpyridinoline 9-1 as a starting material, 6.9 g of solid 9-3 was obtained, with a yield of 41% and a purity greater than 99%. HRMS: m / z (ESI) calcd for C7H3ClN2O[M+H] + 166.99 found: 167.20.

[0201] Example 17: Synthesis of Intermediate 9-5

[0202]

[0203] Referring to steps one and two of Example 2, intermediate 1-3 was replaced with intermediate 9-3, and intermediate 1-4 was replaced with intermediate 9-4, yielding 3.5g of solid 9-5, with a yield of 69% (two steps), a purity greater than 99%, and HRMS: m / z (ESI) calcd for C7H4BrN5O[MH]. - 251.96 found: 252.05.

[0204] Example 18: Synthesis of intermediate 10-3

[0205]

[0206] Following the steps of Example 1, 13.6 g (0.1 mol) of 5-fluoro-2-methylbenzonitrile 10⁻¹ was used as the starting material to yield 7.5 g of solid 10⁻³, with a yield of 50% and a purity greater than 99%. HRMS: m / z (ESI) calcd for C7H₃BrN₂O[M+H] + 151.02 found: 151.20.

[0207] Example 19: Synthesis of Intermediate 10-5

[0208]

[0209] Referring to steps one and two of Example 2, intermediate 1-3 was replaced with intermediate 10-3, and intermediate 1-4 was replaced with intermediate 10-4, yielding 2.7 g of solid 10-5, with a yield of 64% (two steps), a purity greater than 99%, and HRMS: m / z (ESI) calcd for C7H4BrN5O[MH]. - 192.04 found: 191.89.

[0210] Example 20: Synthesis of intermediate 11-5

[0211]

[0212] Referring to steps one and two of Example 2, using (13.2 g, 0.1 mol) 3-formylpyridinium 11-3 as the starting material, and replacing intermediate 1-4 with intermediate 11-4, 4.5 g of solid 11-5 was obtained, with a yield of 25% (two steps), a purity greater than 99%, and HRMS: m / z (ESI) calcd for C7H5N5O[MH]. - 174.05 found: 173.99.

[0213] Example 21: Synthesis of target compound V-1

[0214]

[0215] In a three-necked flask, bromobutane (688.84 mg, 5.03 mmol), iodine granules (100 mg, 0.39 mmol), magnesium strips (100.56 mg, 4.19 mmol), and 10 mL of anhydrous tetrahydrofuran were added. Under nitrogen protection, the mixture was heated to reflux for 12 h. In the same three-necked flask, 189 mg of intermediate 1-5 was dissolved in 15 mL of anhydrous tetrahydrofuran. Under nitrogen protection, the prepared Grignard reagent was injected into the mixture under ice bath conditions. The reaction was carried out at room temperature for 10 h. The crude product was purified by silica gel column chromatography (dichloromethane:methanol = 10:1) to obtain 210 mg of the target compound V-1 with a purity greater than 99%. 1 HNMR (CD3OD, 500MHz) δ: 0.70 (t, 3H, CH3), 0.96~1.25 (m, 6H, CH2), 4.23 (t, 1H, C(OH)H), 7.56 (d, 1H, ArH), 7.75 (dd, 1H, ArH), 7.97 (d, 1H, ArH). HRMS:m / z(ESI)calcd forC 12 H 17 N5O[MH] - 246.14 found: 246.77.

[0216] Example 22: Synthesis of target compound V-2

[0217]

[0218] In a single-necked flask, 100 mg (0.404 mmol) of V-1, 30 mg (1.209 mmol) of sodium hydride, and 10 mL of anhydrous DMF were added. Under nitrogen protection, the mixture was reacted at 45 °C for 0.5 h. Then, 60 mg (0.404 mmol) of iodomethane was injected using a syringe, and the reaction was continued at 38 °C for 4 h. The crude product was purified by silica gel column chromatography (dichloromethane:methanol = 10:1) to obtain 30 mg of the target compound V-2. The purity was greater than 99%. 1 HNMR (CDCl3, 500MHz) δ: 0.71 (t, 3H, CH3), 0.85 ~ 1.35 (m, 6H, CH2), 4.15 (t, 1H, C ( OH)H), 4.65 (s, 3H, N-CH3) 7.77 (d, 1H, ArH), 7.85 (dd, 1H, ArH), 8.01 (d, 1H, ArH). HRMS:m / z(ESI)calcd for C 13 H 19 N5O[M+H] + 262.16 found: 262.35.

[0219] Example 23: Synthesis of target compound V-3

[0220]

[0221] Referring to the steps of Example 21, intermediate 1-5 was replaced with intermediate 2-5 to obtain 120 mg of target compound V-3, with a yield of 62% and a purity greater than 99%. 1 HNMR (CD3OD, 500MHz) δ: 0.85 (t, 3H, CH3), 0.9~1.25 (m, 6H, CH2), 4.13 (t, 1H, C(OH)H), 8.25 (d, 1H, ArH), 8.65 (dd, 1H, ArH), 8.77 (d, 1H, ArH). HRMS:m / z(ESI)calcd for C 13 H 15 F3N4O[MH] - 299.12 found: 299.25.

[0222] Example 24: Synthesis of target compound V-4

[0223]

[0224] Following the steps of Example 22, V-3 was used instead of V-1 to obtain 30 mg of the target compound V-4, with a yield of 65% and a purity greater than 99%.1 HNMR (CDCl3, 500MHz) δ: 0.86 (t, 3H, CH3), 0.95 ~ 1.29 (m, 6H, CH2), 4.37 (t, 1H, C (O H)H), 4.72 (s, 3H, N-CH3), 8.13 (d, 1H, ArH), 8.77 (dd, 1H, ArH), 8.85 (d, 1H, ArH). HRMS:m / z(ESI)calcd for C 14 H 17 F3N4O[M+H] + 315.14 found: 315.26.

[0225] Example 25: Synthesis of target compound V-5

[0226]

[0227] Referring to the steps of Example 21, intermediate 1-5 was replaced with intermediate 3-5 to obtain 80 mg of the target compound V-5, with a yield of 35% and a purity greater than 99%. 1 HNMR (CD3OD, 500MHz) δ: 0.67 (t, 3H, CH3), 0.72~1.26 (m, 6H, CH2), 4.51 (t, 1H, C(OH)H), 8.13 (d, 1H, ArH), 8.35 (dd, 1H, ArH), 8.75 (d, 1H, ArH). HRMS:m / z(ESI)calcd for C 12 H 15 N5O3[MH] - 276.12 found: 276.25.

[0228] Example 26: Synthesis of target compound V-6

[0229]

[0230] Following the steps of Example 22, V-5 was used instead of V-1 to obtain 40 mg of the target compound V-6, with a yield of 78% and a purity greater than 99%. 1 HNMR (CDCl3, 500MHz) δ: 0.69 (t, 3H, CH3), 0.75~1.27 (m, 6H, CH2), 4.25 (s, 3H, N-C H3), 4.37 (t, 1H, C(OH)H), 7.95 (d, 1H, ArH), 8.23 ​​(dd, 1H, ArH), 8.64 (d, 1H, ArH). HRMS:m / z(ESI)calcd for C 12 H 17 N5O[M+H]+ 292.13 found: 292.25.

[0231] Example 27: Synthesis of target compound V-7

[0232]

[0233] Referring to the steps of Example 21, intermediate 1-5 was replaced with intermediate 4-5 to obtain 168 mg of the target compound V-7, with a yield of 69% and a purity greater than 99%. 1 HNMR (CD3OD, 500MHz) δ: 0.69 (t, 3H, CH3), 1.24~1.45 (m, 6H, CH2), 4.57 (t, 1H, C(OH)H), 7.24 (d, 1H, ArH), 7.69 (d, 1H, ArH). HRMS:m / z(ESI)calcd forC 11 H 14 BrN5O[MH] - 310.04 found: 309.88.

[0234] Example 28: Synthesis of target compound V-8

[0235]

[0236] Following the steps of Exercise 22, V-7 was substituted for V-1 to obtain 30 mg of the target compound V-8 with a purity greater than 99%. 1 HNMR (CD3OD, 500MHz) δ: 0.71 (t, 3H, CH3), 0.83~1.25 (m, 6H, CH2), 3.95 (s, 3H, N-CH3), 4.67 (t, 1H, C(OH)H), 7.69 (d, 1H, ArH), 8.13 (d, 1H, ArH). HRMS:m / z(ESI)calcd for C 12 H 16 BrN5O[M+H] + 326.05 found: 325.69.

[0237] Example 29: Synthesis of target compound V-9

[0238]

[0239] Step 1:

[0240] Referring to the steps of Examples 21 and 22, intermediate V-1 is replaced with intermediate 5-5 to obtain intermediate V-9a. 1¹H NMR (500 MHz, CDCl₃) δ: 0.79 (t, 3H, CH₃), 1.24 (m, 6H, CH₂), 7.69 (d, 1H, ArH), 7.77 (dd, 1H, ArH), 7.87 (d, 1H, ArH). Yield 90%, purity >99%, ESI (MH). - =310.

[0241] Step Two:

[0242] Replacing V-1 with V-9a yielded 30 mg of the target compound V-9 with a purity greater than 99%. 1 HNMR (CDCl3, 500MHz) δ: 0.71 (t, 3H, CH3), 0.83~1.25 (m, 6H, CH2), 4.35 (s, 3H, N-C H3), 4.44 (t, 1H, C(OH)H), 7.24 (d, 1H, ArH), 7.75 (dd, 1H, ArH), 7.89 (d, 1H, ArH). HRMS:m / z(ESI)calcd for C 13 H 17 BrN4O[M+H] + 325.06 found: 325.45.

[0243] Example 30: Synthesis of target compound V-10

[0244]

[0245] Step 1:

[0246] Following the steps of Examples 21 and 22, intermediate V-1 was replaced with intermediate 6-5 to obtain intermediate V-10a, with a yield of 85% and a purity greater than 99%. ¹H NMR (500MHz, Methanol-d⁴) values ​​were: δ 7.76 (dd, J = 8.8, 5.7Hz, 1H), 7.42 (dd, J = 9.3, 2.7Hz, 1H), 7.33 (td, J = 8.5, 2.8Hz, 1H), 5.12 (t, J = 6.5Hz, 1H), 1.62 (q, J = 6.8Hz, 2H), 1.48–1.18 (m, 4H), 0.85 (t, J = 7.1Hz, 3H). ESI (MH) - =265.

[0247] Step Two:

[0248] Replacing V-1 with V-10a yielded 45 mg of the target compound V-10 with a purity greater than 99%. 1HNMR (CDCl3, 500MHz) δ: 0.79 (t, 3H, CH3), 0.85 ~ 1.35 (m, 6H, CH2), 4.39 (t, 1H, C (O H)H), 4.56 (s, 3H, N-CH3), 7.35 (d, 1H, ArH), 7.67 (dd, 1H, ArH), 7.98 (d, 1H, ArH). HRMS:m / z(ESI)calcd for C 13 H 17 ClN4O[M+H] + 281.11 found: 281.25.

[0249] Example 31: Synthesis of target compound V-11

[0250]

[0251] Step 1:

[0252] Referring to the steps of Examples 21 and 22, V-1 was replaced with 7-5 to obtain intermediate V-11a, with a yield of 86% and a purity greater than 99%. 1 H NMR (500MHz, Methanol-d4) δ7.74(d,J=8.5Hz,1H),7.69(d,J=2.3Hz,1H),7.59(dd,J=8.5,2.2Hz,1H),5.14(d d,J=7.5,5.3Hz,1H),1.67–1.50(m,2H),1.48–1.36(m,1H),1.35–1.12(m,5H),0.86(t,J=7.0Hz,3H).ESI(MH) - =249.

[0253] Step Two:

[0254] Replacing V-1 with V-11a yielded 35 mg of the target compound V-11 with a purity greater than 99%. 1 HNMR (CDCl3, 500MHz) δ: 0.67 (t, 3H, CH3), 0.75 ~ 1.24 (m, 6H, CH2), 4.36 (s, 3H, N-CH3), 4.41 (t, 1H, C(OH)H), 7.68 (d, 1H, ArH), 7.98 (dd, 1H, ArH), 8.12 (d, 1H, ArH) HRMS: m / z (ESI)calcd for C 13 H 17 FN4O[M+H] + 265.14 found: 265.39.

[0255] Example 32: Synthesis of target compound V-12

[0256]

[0257] Step 1:

[0258] Referring to the steps of Examples 21 and 22, intermediate 8-5 was used instead of V-1, resulting in a yield of 90% and a purity greater than 99%. 1 HNMR (500MHz, CDCl3) δ: 0.90 (t, 3H, CH3, J = 7.5Hz), 1.35-2.05 (m, 6H, CH2), 5.44 (q, 1H, CH, J = 5Hz), 7 .26 (s, 1H, ArH), 7.32 (d, 1H, ArH, J=1Hz), 7.78 (dd, 1H, ArH, J=7.5, 1Hz), 8.02 (d, 1H, ArH, J=7.5Hz). ESI(MH) - =231.

[0259] Step Two:

[0260] Replacing V-1 with V-12a yielded 40 mg of the target compound V-12 with a purity greater than 99%. 1 HNMR (CDCl3, 500MHz) δ: 0.68 (t, 3H, CH3), 0.72 ~ 1.05 (m, 6H, CH2), 3.98 (s, 3H, N-CH3), 4.3 7 (t, 1H, C(OH)H), 7.27 (d, 1H, ArH), 7.48 (m, 1H, ArH), 7.58 (m, 1H, ArH), 7.82 (d, 1H, ArH). HRMS:m / z(ESI)calcd forC 13 H 18 N4O[M+H] + 247.15 found: 247.29.

[0261] Example 33: Synthesis of target compound V-13

[0262]

[0263] Referring to the steps of Example 21, intermediate 1-5 was replaced with intermediate 9-5 to obtain 69 mg of the target compound V-13, with a yield of 38% and a purity greater than 99%. 1HNMR (CD3OD, 500MHz) δ: 0.72 (t, 3H, CH3), 0.85~1.12 (m, 6H, CH2), 4.27 (t, 1H, C(OH)H), 7.69 (d, 1H, ArH), 7.98 (d, 1H, ArH). HRMS:m / z(ESI)calcd forC 11 H 14 ClN5O[MH] - 266.09 found: 265.79.

[0264] Example 34: Synthesis of target compound V-14

[0265]

[0266] Referring to the steps of Example 22, V-1 was replaced with V-13 to obtain 36 mg of the target compound V-14 with a purity greater than 99%. 1 HNMR (CD3OD, 500MHz) δ: 0.69 (t, 3H, CH3), 0.79~1.24 (m, 6H, CH2), 4.29 (t, 1H, C(OH)H), 4.35 (s, 3H, N-CH3), 7.86 (d, 1H, ArH), 8.02 (d, 1H, ArH). HRMS:m / z(ESI)calcd forC 12 H 16 ClN5O[M+H] + 282.10found:282.64.

[0267] Example 35: Synthesis of target compound V-15

[0268]

[0269] Referring to the steps of Example 21, intermediate 1-5 was replaced with intermediate 10-5 to obtain 135 mg of the target compound V-15, with a yield of 64% and a purity greater than 99%. 1 HNMR (CD3OD, 500MHz) δ: 0.76 (t, 3H, CH3), 0.87~1.35 (m, 6H, CH2), 4.39 (t, 1H, C(OH)H), 7.67 (d, 1H, ArH), 8.24 (d, 1H, ArH). HRMS:m / z(ESI)calcd forC 11 H 14 FN5O[MH] - 250.12found:250.15.

[0270] Example 36: Synthesis of target compound V-16

[0271]

[0272] Referring to the steps of Example 22, V-1 was replaced with V-15 to obtain 65 mg of the target compound V-16, with a yield of 64% and a purity greater than 99%. 1 HNMR (CD3OD, 500MHz) δ: 0.75 (t, 3H, CH3), 0.85~1.31 (m, 6H, CH2), 4.46 (s, 3H, N-CH3), 4.55 (t, 1H, C(OH)H), 7.99 (d, 1H, ArH), 8.23 ​​(d, 1H, ArH). HRMS:m / z(ESI)calcd forC 12 H 16 FN5O[M+H] + 266.13 found: 266.36.

[0273] Example 37: Synthesis of target compound V-17

[0274]

[0275] Referring to the steps of Example 21, intermediate 1-5 was replaced with intermediate 11-5 to obtain 116 mg of the target compound V-17, with a yield of 59% and a purity greater than 99%. 1 HNMR (CD3OD, 500MHz) δ: 0.72 (t, 3H, CH3), 0.85~0.98 (m, 6H, CH2), 4.43 (t, 1H, C(OH)H), 7.34 (d, 1H, ArH), 7.86 (m, 1H, ArH), 8.42 (d, 1H, ArH). HRMS:m / z(ESI)calcd for C 11 H 15 N5O[MH] - 232.13 found: 232.15.

[0276] Example 38: Synthesis of target compound V-18

[0277]

[0278] Following the steps of Example 13, V-1 was replaced with V-17 to obtain 62 mg of the target compound V-18, with a yield of 93% and a purity greater than 99%. 1HNMR (CD3OD, 500MHz) δ: 0.67 (t, 3H, CH3), 0.82~1.21 (m, 6H, CH2), 4.35 (s, 3H, N- CH3), 4.45 (t, 1H, C(OH)H), 7.45 (d, 1H, ArH), 7.86 (m, 1H, ArH), 8.55 (d, 1H, ArH). HRMS:m / z(ESI)calcd for C 12 H 17 N5O[M+H] + 248.14 found: 247.99.

[0279] Example 39. Synthesis of intermediate 4-5-3

[0280]

[0281] Referring to the synthesis method of Example 1, replacing raw material 1-1 with 4-5-1 yields intermediate 4-5-3, ESI(M+H). + =210.

[0282] Example 40. Synthesis of intermediate 4-5-5

[0283]

[0284] Referring to the synthesis method of Example 2, raw materials 1-3 were replaced with 4-5-3 to obtain intermediate 4-5-5, ESI(MH). - =251.

[0285] Example 41. Synthesis of target compound V-19

[0286]

[0287] Referring to the steps of Example 21, intermediate 1-5 was replaced with intermediate 8-5, and bromopropane was replaced with bromobutane to obtain 160 mg of the target compound V-19, with a yield of 83% and a purity greater than 99%. 1 HNMR (500MHz, CDCl3) δ: 0.82 (t, 3H, CH3, J = 7.5Hz), 1.25-1.63 (m, 4H, CH2), 5.01 (q, 1H, CH, J = 5Hz), 7.39 (td, 1 H, ArH, J=7.5, 1Hz), 7.52 (td, 1H, ArH, J=7.5, 1Hz), 7.60 (dd, 1H, ArH, J=7.5, 1Hz), 7.67 (td, 1H, ArH, J=7.5Hz). ESI(MH) - =217.

[0288] Example 42. Synthesis of target compound V-20

[0289]

[0290] Referring to the steps of Example 21, intermediate 1-5 was replaced with intermediate 8-5, and bromopentane was replaced with bromobutane, yielding 203 mg of the target compound V-20, with a yield of 93%. 1 HNMR (500MHz, CDCl3) δ: 0.85-1.62 (m, 8H, CH2), 1.37 (m, 3H, CH3), 5.07 (t, 1H, CH, J = 6.5H z), 7.43 (td, 1H, ArH, J=8, 1.5Hz), 7.58 (t, 2H, ArH, J=7.5Hz), 7.43 (d, 1H, ArH, J=7.5Hz). ESI(MH) - =245.

[0291] Example 43. Synthesis of target compound V-21

[0292]

[0293] Referring to the steps of Example 21, intermediate 1-5 was replaced with intermediate 5-5, and bromopentane was replaced with bromobutane to obtain the target compound V-21 in 91% yield. 1 H NMR(500MHz, Methanol-d4)δ7.74(d,J=8.5Hz,1H),7.69(d,J=2.3Hz,1H),7.62–7.56(m,1H),5.14(dd,J=7.7,5.2 Hz,1H),1.69–1.52(m,2H),1.38(tdd,J=10.9,6.0,2.6Hz,1H),1.33–1.19(m,3H),0.85(t,J=7.1Hz,3H).ESI(MH) - =323.

[0294] Example 44. Synthesis of target compound V-22

[0295]

[0296] Referring to the steps of Example 21, intermediate 1-5 was replaced with intermediate 6-5, and bromopentane was replaced with bromobutane to obtain the target compound V-22 in 95% yield. 1H NMR (500MHz, Methanol-d4) δ7.83(d,J=2.1Hz,1H),7.74(dd,J=8.5,2.1Hz,1H),7.68(d,J=8.4Hz,1H),5.13(dd,J=7.7,5 .2Hz,1H),1.69–1.51(m,2H),1.41(dtd,J=11.9,7.1,5.7,2.8Hz,1H),1.35–1.08(m,4H),0.86(t,J=7.0Hz,3H).ESI(MH) - =279.

[0297] Example 45: Synthesis of target compound V-23

[0298]

[0299] Referring to the steps of Example 21, intermediate 1-5 was replaced with intermediate 7-5, and bromopentane was replaced with bromobutane to obtain the target compound V-23 in 89% yield, ESI (MH). - =263.

[0300] Example 46: Synthesis of target compound V-24

[0301]

[0302] Referring to the steps of Example 21, intermediate 1-5 was replaced with intermediate 4-5-5 to obtain the target compound V-24. 1 HNMR(500MHz,Methanol-d4)δ7.92(d,J=2.0Hz,1H),7.60(dd,J=8.3,2.1Hz,1H),7.55(d,J=8.3H z,1H),5.16(dd,J=8.3,4.4Hz,1H),1.68–1.49(m,2H),1.46–1.14(m,3H),0.85(t,J=7.2Hz,3H). ESI(MH) - =310.

[0303] Example 47: Synthesis of target compound V-25

[0304]

[0305] Referring to the steps of Example 21, replacing 1-5 with 4-5-5 yields the target compound V-25, ESI(MH). - =323.

[0306] Example 48: Synthesis of target compound V-26

[0307]

[0308] Referring to the steps of Example 21, 5-5 was used instead of 1-5, and bromopropane was used instead of bromobutane to obtain the target compound V-26, ESI(MH). - =295.

[0309] Example 49: Synthesis of target compound V-27

[0310]

[0311] Referring to the steps of Example 21, 6-5 was used instead of 1-5, and bromopropane was used instead of bromobutane to obtain the target compound V-27, ESI(MH). - =251.

[0312] Example 50: Synthesis of target compound V-28

[0313]

[0314] Referring to the steps of Example 21, 7-5 was used instead of 1-5, and bromopropane was used instead of bromobutane to obtain the target compound V-28, ESI(MH). - =235.

[0315] Example 51: Synthesis of target compound V-29

[0316]

[0317] Referring to the steps of Example 22, V-1 was replaced with V-26 to obtain the target compound V-29 in 81% yield, ESI (MH). - =309.

[0318] Example 52: Synthesis of target compound V-30

[0319]

[0320] Referring to the steps of Example 22, V-1 was replaced with V-27 to obtain the target compound V-30, ESI(MH). - =265.

[0321] Example 53: Synthesis of target compound V-31

[0322]

[0323] Referring to the steps of Example 22, V-1 was replaced with V-28 to obtain the target compound V-31, ESI(MH). - =249.

[0324] Example 54: Synthesis of target compound V-32

[0325]

[0326] Referring to the steps of Example 22, V-1 was replaced with V-21 to obtain the target compound V-32, ESI(MH). - =337.

[0327] Example 55: Synthesis of target compound V-33

[0328]

[0329] Referring to the steps of Example 22, V-1 was replaced with V-22 to obtain the target compound V-33, ESI(MH). - =293.

[0330] Example 56: Synthesis of target compound V-34

[0331]

[0332] Referring to the steps of Example 22, V-1 was replaced with V-23 to obtain the target compound V-34, ESI(MH). - =277.

[0333] Example 57: Synthesis of target compound V-35

[0334]

[0335] Referring to the steps of Example 22, V-1 was replaced with V-20 to obtain the target compound V-35, ESI(MH). - =259.

[0336] Example 58: Synthesis of target compounds V-36 and V-37

[0337] Using V-9a as a starting material, compounds V-36 and V-37 were obtained by chiral column chromatography.

[0338] Example 59: Synthesis of target compounds V-38 and V-39

[0339] Using V-10a as a starting material, compounds V-38 and V-39 were obtained by chiral column chromatography.

[0340] Example 60: Synthesis of target compounds V-40 and V-41

[0341] Using V-11a as a starting material, compounds V-40 and V-41 were obtained by chiral column chromatography.

[0342] Example 61: Synthesis of target compounds V-42 and V-43

[0343] Using V-21 as a raw material, compounds V-42 and V-43 were obtained by chiral column chromatography.

[0344] Example 62: Synthesis of target compounds V-44 and V-45

[0345] Using V-22 as a raw material, compounds V-44 and V-45 were obtained by chiral column chromatography.

[0346] Example 63: Synthesis of target compounds V-46 and V-47

[0347] Using V-23 as a starting material, compounds V-46 and V-47 were obtained by chiral column chromatography.

[0348] Example 64: Synthesis of target compounds V-48 and V-49

[0349] Using V-24 as a raw material, compounds V-48 and V-49 were obtained by chiral column chromatography.

[0350] Example 65: Synthesis of target compounds V-50 and V-51

[0351] Using V-25 as a raw material, compounds V-50 and V-51 were obtained by chiral column chromatography.

[0352] Example 66: Inhibitory effect of the compound disclosed in this invention on ADP-induced platelet aggregation in vitro.

[0353] Washed platelet samples without any reagents or drugs were used as a blank control group; washed platelet samples treated with ADP and dimethyl sulfoxide (DMSO) were used as negative controls; and washed platelet samples treated with ADP and the marketed drug butylphthalide (NBP) were used as positive controls. The inhibitory effect of the compounds obtained in this invention on ADP-induced platelet aggregation was determined using an in vitro platelet aggregation assay.

[0354] The experimental method for inhibiting ADP-induced platelet aggregation by the compounds of this invention is as follows:

[0355] Experimental materials:

[0356] Drug: Test drug: The monomeric compound obtained in this invention. Positive control: Butylphthalide, purchased from Shanghai Bide Pharmaceutical Technology Co., Ltd. Purity >95%.

[0357] Experimental animals: SPF-grade male SD rats. They were housed in stainless steel wire cages with a volume of 500×350×200 mm (length×width×height), and no more than 5 rats were kept in each cage. During the experiment, no animals of other species were housed in the same room area. License number for the use of experimental animals: SYXK(Zhe)2012-0178. The temperature was strictly controlled at 18-26°C, the humidity was 40%-70%, the daily temperature difference was not more than 4°C, the ventilation rate was >8 times / hour, and the light was controlled with a 12-hour light / 12-hour dark cycle (light from 8:00 to 20:00). The experimental animals drank water freely from water bottles and ate freely.

[0358] Method for obtaining washed platelet samples: SD rats were anesthetized by intraperitoneal injection of 10% chloral hydrate at 0.3 ml / 100 g. Blood was collected from the abdominal aorta, and the blood volume collected from each animal each time was about 10 ml. The blood was placed in a centrifuge tube containing citrate glucose anticoagulant (ACD). The blood and ACD were mixed evenly at a volume ratio of 9:1. Centrifuged at 120 g and 25°C for 20 min to obtain the supernatant, which was platelet-rich plasma (PRP). The PRP was diluted with ACD to prepare washed platelets, and the blood and ACD were mixed evenly at a volume ratio of 1:3, then centrifuged at 800 g and 25°C for 10 min. The precipitate was platelets. Carefully aspirate the upper supernatant and transfer it to a new centrifuge tube. Centrifuge the lower liquid at 800 g for 10 minutes, and aspirate the upper liquid as platelet-poor plasma (PPP) for standby. The precipitate was resuspended with platelet resuspension solution (Tyrode's buffer). After counting with a hemocytometer, the platelets were diluted to 200×10 9 cells / L with the resuspension solution Tyrode's buffer.

[0359] In vitro platelet aggregation assay: Blank control group: Washed platelet samples (290 μL) were incubated at room temperature for 3 min in a disposable sample cup equipped with a disposable stir bar, then placed in the incubation well of a Prisen four-channel platelet aggregation analyzer and incubated at 37°C for 3 min. After zeroing with PPP, 10 μL of the inducer ADP was added (ADP system concentration was 10 μM; the blank control group was treated with 10 μL of physiological saline). Platelet aggregation rate was measured, and the maximum platelet aggregation rate (%) within 300 s was recorded. The platelet aggregation-inducing effect of ADP was confirmed. Negative control group: Washed platelet sample (288 μl) was placed in a disposable sample cup equipped with a disposable stir bar, 2 μL of DMSO was added, and the sample was incubated at room temperature for 3 min. Then, it was placed in the incubation well of the Prisen four-channel platelet aggregation analyzer and incubated at 37°C for 3 min. After zeroing with PPP, 10 μL of ADP (10 μM concentration in the ADP system) was added, and the platelet aggregation rate was measured. The maximum platelet aggregation rate (%) within 300 s was recorded. Each group was repeated three times. Positive control group and experimental group: 288 μl of washed platelet sample was placed in a disposable sample cup equipped with a disposable stir bar. 2 μL of butylphthalide or the compound obtained in this invention (system concentration of compound: 0.1 mM) was added. The sample was incubated at room temperature for 3 min, then placed in the incubation well of a Prisen four-channel platelet aggregation assay instrument and incubated at 37°C for 3 min. After zeroing with PPP, 10 μL of the inducer ADP was added (system concentration of ADP: 10 μM), and the platelet aggregation rate was measured. The maximum platelet aggregation rate (%) within 300 s was recorded. Each group was repeated three times. Inhibition rate calculation formula:

[0360]

[0361] The experimental data were analyzed using data analysis software provided by Graphpad Prism; a one-way analysis was used here. The data is shown in Table 1.

[0362] Table 1. Inhibitory effect of the compounds disclosed in this invention on ADP-induced platelet aggregation.

[0363]

[0364]

[0365] +++ indicates an inhibition rate greater than 30%.

[0366] ++ indicates an inhibition rate greater than 20% and less than 30%.

[0367] + indicates an inhibition rate greater than 10% and less than 20%.

[0368] NS indicates a value less than 10%.

[0369] As shown in Table 1, the 0.1 mM compound disclosed in this invention showed no significant difference in the inhibition rate of ADP-induced in vitro platelet aggregation compared to the positive control drug butylphthalide, indicating that the compound disclosed in this invention and the positive control drug have comparable inhibitory activity on platelet aggregation.

[0370] Example 67: Protective effect of the compounds disclosed in this invention on primary cortical neurons of OGD / R fetal rats

[0371] Normal neurons that did not undergo OGD / R modeling were used as the control group, neurons that underwent OGD / R modeling but were not treated with drugs were used as the model group, neurons that underwent OGD / R modeling and were treated with the marketed drugs butylphthalide (NBP) and BZP were used as the positive control, and neurons that underwent OGD / R modeling and were treated with the compounds of this invention were used as the experimental group. The protective effect of the compounds obtained in this invention on primary cortical neurons of fetal rats modeled with OGD / R was determined by the CCK8 assay.

[0372] The pharmacological experimental method for the neuronal cell protective effect of the compounds of this invention in OGD / R modeling is as follows:

[0373] Experimental materials:

[0374] Drug: Test drug: The monomeric compound obtained in this invention. Positive control drugs: Butylphthalide and BZP were purchased from Shanghai Bide Pharmaceutical Technology Co., Ltd. Purity >95%.

[0375] Cell line: Neurons in the cerebral cortex of ICR fetal rats on day 15.

[0376] Culture medium and additives: Neurobasal (NB culture medium), B27 additive, GlutaMax additive, sugar-free DMEM, FBS, manufactured by Gibco, USA.

[0377] Culture medium preparation method:

[0378] NBM / B27 medium - low glutamine version: If using a 50mL centrifuge tube, add 45mL NB medium, 0.9mL B27 additive, 1% penicillin-antibiotic, and 11.25μL Glutamax additive.

[0379] NBM / B27 medium - high glutamine version: If using a 50mL centrifuge tube, add 45mL NB medium, 0.9mL B27 additive, 1% penicillin and antibiotic, and 27μL Glutamax additive.

[0380] Drug preparation method: Dissolve the drug in DMSO to prepare a stock solution of the corresponding concentration, and then dilute it with culture medium at a certain ratio.

[0381] Cell acquisition and culture:

[0382] The pregnant mouse was euthanized by cervical dislocation, placed in a supine position, and its uterus was removed through an abdominal incision. The fetus was then placed in an HBSS (Hepatocellular Carcinoma Spectrostomy System), and its brain was removed and the surface vascular membrane was separated using surgical scissors and forceps.

[0383] Digest cells with trypsin and DNase at 37°C for 25-30 min. After digestion, terminate digestion with 10% FBS + DMEM, then transfer to DMEM. Use pipette tips of decreasing diameter to pipette the cells until there is no resistance, collect the supernatant and centrifuge. Discard the supernatant after centrifugation, dilute the pellet with medium (NBM / B27 medium - high glutamine version) to 500,000 cells / mL, and seed into 96-well plates (100,000 cells / well, 100 μL / well).

[0384] The obtained cells were incubated in a 37°C, 5% CO2 cell culture incubator, with half the medium changed every three days (NBM / B27 medium - low glutamine version), and the drug was administered 2 hours before OGD on the seventh day.

[0385] Drug administration: The compound was dissolved in dimethyl sulfoxide (DMSO) to prepare a 20,000 μM stock solution, which was then diluted with culture medium (NBM / B27 medium-low glutamine version) to obtain culture media containing the drug (20 μM and 2 μM). 50 μL of the culture medium was removed from each well, and 50 μL of the culture medium containing the drug was added to each well to a final concentration of 10 μM. The 1 μM solutions were incubated together in a 37°C, 5% CO2 cell culture incubator for 2 hours. The blank group and the model group did not have their medium changed.

[0386] Modeling:

[0387] ODG: After 2 hours, the cell culture medium was replaced with sugar-free DMEM (pre-placed in an anaerobic chamber for 20 minutes to remove O2, and washed three times), and placed in an anaerobic chamber (95% N2 + 5% CO2) and cultured at 37°C for 2 hours.

[0388] R: After 2 hours, remove from the anaerobic chamber and replace the cell culture medium with NBM / B27 medium - low glutamine version. Incubate at 37°C, 5% CO2 in a cell culture incubator for 24 hours.

[0389] Cell viability was determined using the CCK8 kit: 10 kits of cell viability assay solution were added to each well and incubated in a 37°C, 5% CO2 cell culture incubator for 2 hours. The absorbance of each well was measured at 450 nm.

[0390] The formula for calculating cell viability is: Cell viability % = (OD value of drug-treated group - OD value of background in blank wells) / (OD value of control group cells - OD value of background in blank wells) / (OD value of background in blank wells) / (OD value of background in blank wells)

[0391] The experimental data were analyzed using data analysis software provided by Graphpad Prism; a one-way analysis was employed here. The data are shown in Table 2. Figure 2 result.

[0392] Table 2. Protective effects of compounds on primary cortical neurons in fetal rats induced by OGD / R.

[0393]

[0394]

[0395] Note: +: 0-0.2, ++: 0.2-0.4, +++: 0.4 or more

[0396] t-test, compared with the control group, ####: P<0.0001.

[0397] t-test, compared with the model group, ****: P<0.0001, ***: P<0.001, *: P<0.05.

[0398] As shown in Table 2, there were significant differences between the control group and the model group in this pharmacological experiment, indicating that the OGD / R model was successful and the cell survival rate decreased after OGD / R treatment. There were also significant differences between the model group and each drug administration group, indicating that the positive drugs NBP and BZP and the target compound all have protective effects on nerve cells and can improve their survival rate.

[0399] Depend on Figure 2 It can be seen that the cell viability of the target compound group was higher than that of the NBP and BZP groups. Among them, there was no significant difference between the V-9a-1μM group and the NBP-10μM and BZP-10μM groups; however, the 10μM group of the present invention showed a significant difference compared with the NBP-10μM and BZP-10μM groups, and was significantly higher than the NBP-10μM and BZP-10μM groups. It can be considered that the compound of the present invention exhibits a high neuroprotective effect at low concentrations.

[0400] Example 68: Protective effect of the compound disclosed in this invention against OGD / R damage in neuroblastoma cells (N2A).

[0401] Neuroblastoma cells were induced to differentiate into neuron-like cells and then used for OGD / R modeling. After 4 hours of oxygen-glucose deprivation, reperfusion was achieved by adding DMEM medium containing serum and glucose. Subsequent analysis was performed on cells 24 hours after reperfusion. The experiment consisted of 8 groups: Control group, model group, positive control drug NBP-10μM group, V-9a 0.1μM group, V-9a 1μM group, V-9a 10μM group, and V-9a 100μM group. The corresponding drug for each group was added to the cell culture medium during reperfusion.

[0402] Cell viability was assessed using the Cell Counting Kit-8 (CCK-8), and the results are shown below. Figure 3 The results showed that cell viability was significantly reduced after OGD / R modeling, while both V-9a and NBP significantly improved the activity of neuron-like cells after ischemia and hypoxia, with the V-9a 10μM group showing a better protective effect than NBP. The release of cytoplasmic LDH indicated loss of cell membrane integrity, representing cell death. LDH release was significantly increased in the OGD / R modeling group, while both V-9a and NBP significantly reduced LDH release from neuron-like cells.

[0403] Example 69: Properties of the compounds disclosed in this invention in oral pharmacokinetics in rats

[0404] Experimental design: Three male SD rats were used to conduct the experiment according to the table below.

[0405]

[0406] Sample collection: 0.10 mL of blood was collected from each animal via the orbital cavity, anticoagulated with EDTAK2, at 15 min, 30 min, 1 h, 2 h, 4 h, 6 h, and 8 h after administration of the test substance. Blood samples were placed on ice after collection and centrifuged within 30 minutes to separate the plasma (centrifugation conditions: 5000 rpm, 10 min, 4 °C). The plasma was stored at -80 °C before analysis.

[0407] LC-MS / MS conditions:

[0408] Liquid chromatography method: Column: ACQUITY BEH C18 2.1x50mm 1.7μm

[0409] Mobile phase A: 0.1% formic acid in water; Mobile phase B: acetonitrile; Flow rate: 0.35 mL / min

[0410] Changes in mobile phase ratio over time:

[0411]

[0412] Injection volume: 10 μL

[0413] Mass spectrometry method: capillary voltage: 3.5 kV, desolvation gas temperature: 500 ℃, desolvation gas flow rate: 1000 L / Hr, cone gas flow rate: 50 L / Hr.

[0414] Drug concentration standard curve preparation: The stock solution of the test compound was diluted with 50% methanol-water to prepare standard working solutions containing concentrations of 40, 100, 200, 400, 1000, 2000, 4000, 10000, and 20000 ng / mL for each compound, and quality control working solutions containing concentrations of 120, 1200, and 12000 ng / mL. 2.50 μL of standard curve working solution and quality control working solution were added to 47.5 μL of blank matrix to prepare drug concentrations of 2... Standard curves with concentrations of 0.00, 5.00, 10.00, 20.00, 50.00, 100.00, 200.00, 500.00, and 1000.00 ng / mL, and quality control samples with concentrations of 6.00, 60.00, and 600.00 ng / mL, were added to 200 μL of acetonitrile (containing 1 ng / mL of loratadine). After vortexing for 3 min, the samples were centrifuged at 20000 rcf and 4 °C for 10 min. The supernatant was then used for LC-MS / MS analysis.

[0415] Blood drug concentration determination: Take 50 μL of plasma sample, add 200 μL of acetonitrile (containing loratadine 1 ng / mL), vortex for 3 min, centrifuge at 20000 rcf, 4℃ for 10 min, and collect the supernatant for LC-MS / MS analysis. The blood drug concentration is calculated using the standard curve, and Cmax (ug / L) and AUC are calculated using Das 2.0. 0-t (hr*ng / mL), as shown in Table 3.

[0416] Table 3. Partial pharmacokinetic parameters of the test compounds (IG 10 mg / kg)

[0417]

[0418] Note: NA: Below the detection limit, cannot be calculated.

[0419] As shown in Table 3, when the compounds disclosed in this invention are administered orally, they achieve high plasma drug concentrations and exposure levels in SD rats, demonstrating significant advantages over butylphthalide and BZP. Experimental results indicate that the compounds disclosed in this invention can be ideal candidate molecules for oral drug administration.

[0420] Example 70: Pharmacological effects of the compounds disclosed in this invention in animal models of stroke.

[0421] A focal cerebral ischemia animal model was established using the suture occlusion method with transient middle cerebral artery occlusion to test the in vivo anti-stroke activity of V-9a. SPF-grade C57 mice, 10 weeks old, healthy males, weighing 22-25g, were used in the experiment. The experiment was divided into five groups: MCAO + Saline group (1ml saline), MCAO + NBP (10mg / kg, IV), MCAO + V-9a (3mg / kg, IV), MCAO + V-9a (10mg / kg, IV), and MCAO + V-9a (30mg / kg, IV), with 10 animals in each group. Animals in each group received a single intravenous injection of the drug immediately after MCAO reperfusion. 24 hours later, behavioral scores, laser speckle imaging to observe cerebral blood flow recovery, and TTC staining of brain tissue were performed.

[0422] Compared with the model group, both NBP and V-9a reduced neurological scores and significantly improved behavioral symptoms in mice. TTC staining showed a significant reduction in cerebral infarction volume after V-9a administration, and the ischemic protective effect of V-9a was dose-dependent. Furthermore, the cerebral infarction volume in the 30 mg / kg V-9a group was significantly lower than that in the 10 mg / kg NBP group. This suggests that high-dose V-9a has a superior protective effect against cerebral ischemia-reperfusion injury compared to NBP.

[0423] In summary, the tetrazolium derivatives involved in this invention have broad prospects for drug application in the prevention and treatment of cardiovascular and cerebrovascular diseases, the improvement of cardiovascular and cerebrovascular circulation disorders, or the prevention of thrombosis.

Claims

1. A tetrazolium derivative, characterized in that, The tetrazolium derivative is 1-(6-bromo-2-(1H-tetrazol-5-yl)pyridin-3-yl)pent-1-ol or a pharmaceutically acceptable salt thereof.

2. The tetrazolium derivative according to claim 1, characterized in that, The pharmaceutically acceptable salt is a salt formed with one or more of the following bases: sodium hydroxide, potassium hydroxide, lithium hydroxide, sodium methoxide, potassium methoxide, sodium ethoxide, potassium ethoxide, triethylamine, and tert-butylamine.

3. The tetrazolium derivative according to claim 1, characterized in that, The tetrazolium derivatives include: 1-(6-bromo-2-(1H-tetrazol-5-yl)pyridin-3-yl)pentan-1-ol sodium salt 1-(6-bromo-2-(1H-tetrazol-5-yl)pyridin-3-yl)pent-1-ol potassium salt 1-(6-bromo-2-(1H-tetrazol-5-yl)pyridin-3-yl)pent-1-ol lithium salt 1-(6-bromo-2-(1H-tetrazol-5-yl)pyridin-3-yl)pent-1-ol triethylamine salt 1-(6-bromo-2-(1H-tetrazol-5-yl)pyridin-3-yl)pent-1-ol tert-butylamine salt.

4. A method for preparing the tetrazolium derivative according to claim 1, characterized in that, Compound 4-3 reacts with an acyl protecting agent to give compound 4-4. Compound 4-4 then reacts with sodium azide, and hydrochloric acid is added to remove the protecting group and eliminate the remaining sodium azide, giving compound 4-5. Compound 4-5 reacts with a Grignard reagent to give 1-(6-bromo-2-(1H-tetrazol-5-yl)pyridin-3-yl)pentan-1-ol. 。 5. The method for preparing the tetrazolium derivative according to claim 4, characterized in that, The acyl protecting agent is ethylene glycol, and the deprotecting agent is hydrochloric acid.

6. The method for preparing the tetrazolium derivative according to claim 4, characterized in that, The reaction temperature of compound 4-3 with the acyl protecting agent is 110~130℃; the reaction temperature of compound 4-4 with sodium azide is 90~110℃.

7. A pharmaceutical composition, characterized in that, The pharmaceutical composition comprises at least one active ingredient and one or more pharmaceutically acceptable carriers or excipients, wherein the active ingredient is a tetrazolium derivative as described in claim 1.

8. The use of a tetrazolium derivative of claim 1 in the preparation of a medicament for the prevention and treatment of cardiovascular and cerebrovascular diseases, the improvement of cardiovascular and cerebrovascular circulation disorders, or the prevention of thrombosis.

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