A benzotriazole compound, a preparation method and application thereof

Abstract

The application discloses a benzotriazole compound, a preparation method and application thereof, and belongs to the technical field of medicinal chemistry. The compound has structures as shown in general formula (I), (II) and (III). A fragment containing an amide bond substituted amino acid ester or amino acid structure is introduced into the system based on benzotriazole to synthesize a series of benzotriazole compounds containing the amino acid ester or amino acid structure. The compound has a good inhibiting effect on plant pathogenic viruses, and especially has a good inhibiting effect on plant pathogenic viruses such as tobacco mosaic virus.

Description

Technical Field

[0001] This invention relates to the field of medicinal chemistry, and in particular to a benzotriazole compound, its preparation method, and its application. Background Technology

[0002] Plant cancers caused by viruses, pathogens, fungi, bacteria, and oomycetes severely and persistently restrict crop yield and quality. Indeed, plant viruses are representative pathogens of crops. Plant diseases caused by viruses can devastatingly reduce agricultural production and deform the appearance of plants. Plant viruses reside in host cells, which depend on the host for the materials and environment to replicate. Because plants lack a complete immune system, they are vulnerable to external environmental influences, making plant viral damage ubiquitous.

[0003] Tobacco mosaic virus (TMV) is one of the most difficult viruses to manage infecting tobacco plants, and it is the earliest discovered and most studied plant virus. It is known to infect at least 400 species, causing irreparable leaf mosaic in subsequent growth. Among commercial antiviral agents, ribavirin is primarily used to protect plants from viral infection. However, ribavirin's antiviral efficacy is less than 50%, and its field cure rate is unsatisfactory. Furthermore, environmental changes lead to potential viral mutations and enhanced resistance, thus plant antiviral agents rarely produce effective and sustainable long-term suppression of viruses.

[0004] The following are the research advances on the bioactivity based on triazoles, amino acids, or amide bonds:

[0005] In 2013, Li et al. [Li,YD;Mao,WT;Fan,ZJ;Ji,XT;Hua,XW;Zong,GN;Li,FY;Liu,CL;Yu,JHS] synthesized compounds 42, 43, 44, and 45 with a 1,2,4-triazole structure. In vivo assays using the half-leaf method showed that at a concentration of 100 μg / mL, compounds 42, 43, 44, and 45 exhibited inhibition rates of 45.5%, 51.9%, 50.8%, and 40.7% against tobacco mosaic virus, respectively. Compounds 43 and 44 showed superior inhibition compared to the control drug ningnanmycin (45.0%).

[0006] In 2015, Petrova et al. [Petrova,kt; Potewar,TM; Correia-da-Silva,P.; TeresaBarros,M.; Calhelhac,RC; Ciricd,A.; Sokovic d,M.; Ferreira,FR Antimicrobialand cytotoxic activities of 1,2,3-triazole-sucrose derivatives[J]. Carbohydrate Research,2015,417:66-71] designed and synthesized a series of glycocyclic compounds containing 1,2,3-triazole. Through antibacterial, antifungal, and cytotoxicity tests, they found that most of the target compounds showed good inhibitory activity against various clinically and food contaminants of important microorganisms. Among them, compound 5 had the highest activity, with a minimum inhibitory concentration between 1.1 and 4.4 μM and a minimum bactericidal concentration between 2.2 and 8.4 μM, and the cytotoxicity was controlled within a reasonable range.

[0007] In 2017, Wang et al. [Wang, X.; Dai, ZC; Chen, YF; Cao, LL; Yan, W.; Li, SK; Wang, JX; Zhang, ZG; Ye, YGS] used 1,2,3-triazole as the parent compound to perform structural derivatization. They screened the activity of synthesized 1,2,3-triazole compounds by inhibiting the hyphal growth of pathogenic plant fungi in vitro. They found that when compounds contained both 1,2,3-triazole and acyl structural skeletons, the activity of the compounds increased significantly. For example, 6ad showed strong activity against four plant pathogenic fungi: Rhizoctonia solani, Sclerotinia sclerotiorum, Fusarium graminearum, and Bacillus oryzae. 50 The concentrations were 0.18, 0.35, 0.37, and 2.25 mg / mL, respectively. In vivo experiments showed that the primary target of 6ad was the rice sheath.

[0008] In 2020, Garg et al. [Garg, A.; Borah, D.; Trivedi, P.; Gogoi, D.; Chaliha, AK; Ali, AA; Chetia, D.; Chaturvedi, V.; Sarma, DA. Simple Work-Up-free, Solvent-free Approach to Novel Amino Acid Linked 1,4-Disubstituted 1,2,3-Triazoles as Potent Antituberculasis Agents[J]. ACS Omega 2020, 5, 29830-29837] synthesized novel amino acid molecules containing 1,2,3-triazole structures using 1,8-diazabicyclic acetate ionic liquids. Anti-tuberculosis and antibacterial activity analyses showed that these molecules exhibited good antibacterial activity and considerable affinity for the DprE1 target protein of Mycobacterium tuberculosis in experimental studies. Cytotoxicity assays and computer analysis indicated that these synthetic molecules have potential application value.

[0009] In 2020, Kim et al. [Kim, K.; Kwon, H.; Barinka, C.; Motlova, L.; Nam, S.; Choi, D.; Ha, H.; Nam, H.; Son, SH; Minn, I.; Pomper, MG; Yang, X.; Kutil, Z.; Byun, Y. Novel β-and γ-Amino Acid-Derived Inhibitors of Prostate-Specific Membrane Antigen[J]. J. Med. Chem. 2020, 63, 3261-3273] designed and synthesized a series of compounds containing the Lys-urea-Glu motif targeting prostate-specific membrane antigen (PSMA). Structure-activity relationship studies revealed that compound 13c, a β-amino acid analog with an (R) configuration, exhibited the most potent PSMA inhibitory activity, with an IC50 value of 3.97 nM. The X-ray crystal structure of PSMA-13c composites provides a mechanistic basis for the stereochemistry of PSMA and guides the development of PSMA inhibitors.

[0010] In 2021, Shao et al. [Shao, WB; Wang, PY; Fang, ZM; Wang, JJ; Guo, DX; J, J.; Zhou, X.; Qi, PY; Liu, LW; Yang, S. Synthesis and Biological Evaluation of 1,2,4-Triazole Thioethers as Both Potential Virus Factor Inhibitors against Plant Bacterial Diseases and Agrictural Antiviral Agents against TobaccoMosaic Virus Infections[J]. J. Agric. Food Chem. 2021, 69, 15108-15122] prepared and screened several types of 1,2,4-triazole thioethers with amide bonds. Bioassay results showed that 1,2,4-triazole thioether A10 with a specific N-(3-nitrophenyl)acetamide fragment exhibited extremely strong biological activity against Xanthomonas oryzae. The EC50 value of oryzae (Xoo) was 5.01 μg / mL. In vivo anti-Xoo assays showed that compound A10 had a control efficacy (54.2-59.6%) superior to copper thiamethoxam and dimethylthiazole (38.1-44.9%). Furthermore, compound A10 exhibited significant antiviral activity against tobacco mosaic virus (TMV), with therapeutic and protective activities of 54.6% and 76.4%, respectively, comparable to the control agent ningnanmycin (55.2% and 60.9%).

[0011] However, traditional antiviral agents suffer from poor stability and unsatisfactory field efficacy, and their large-scale use also poses potential safety risks. Therefore, there is an urgent need to develop new green pesticides with high activity and novel mechanisms of action. Summary of the Invention

[0012] In order to find pesticide candidates with high antiviral activity, this invention synthesizes a series of benzotriazole compounds with amide bond substitution or amino acid structure based on benzotriazole, and investigates their bioactivity against plant viruses, providing an important scientific basis for the research and development and creation of new antiviral agents.

[0013] This invention provides a benzotriazole compound with the general structural formulas shown in formulas (I), (II), and (III):

[0014]

[0015] Wherein, R1 is selected from one or more of hydrogen, deuterium, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted alkoxy, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl.

[0016] R2 is selected from one or more of hydrogen, deuterium, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted alkoxy, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl.

[0017] R3 is selected from one or more of hydrogen, deuterium, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted alkoxy, substituted or unsubstituted cycloalkyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl.

[0018] Further, R1 is selected from one or more of hydrogen, deuterium, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, substituted or unsubstituted C6-C15 aryl, substituted or unsubstituted C6-C10 heteroaryl, wherein the substitution refers to being substituted by one or more of C1-C6 alkyl, C1-C6 alkoxy, amino, hydroxyl, halogen, nitro, and trifluoromethyl;

[0019] R2 is selected from one or more of hydrogen, deuterium, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, substituted or unsubstituted C6-C15 aryl, substituted or unsubstituted C6-C10 heteroaryl, wherein the substitution refers to being substituted by one or more of C1-C6 alkyl, C1-C6 alkoxy, amino, hydroxyl, halogen, nitro, and trifluoromethyl.

[0020] R3 is selected from one or more of hydrogen, deuterium, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, substituted or unsubstituted C6-C15 aryl, substituted or unsubstituted C6-C10 heteroaryl, wherein the substitution refers to being substituted by one or more of C1-C6 alkyl, C1-C6 alkoxy, amino, hydroxyl, halogen, nitro, and trifluoromethyl.

[0021] Further, R1 is selected from hydrogen, deuterium, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, 1,1-dimethyl, 1,5-dimethylhexyl, 1,1-diethanolyl, propenyl, allyl, methoxy, ethoxy, propoxy, butoxy, substituted or unsubstituted phenyl, substituted or unsubstituted benzyl, wherein the substitution refers to being substituted by one or more of C1-C6 alkyl, C1-C6 alkoxy, amino, hydroxyl, halogen, nitro, and trifluoromethyl;

[0022] R2 is selected from hydrogen, deuterium, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, 1,1-dimethyl, 1,5-dimethylhexyl, 1,1-diethanolyl, propenyl, allyl, methoxy, ethoxy, propoxy, butoxy, substituted or unsubstituted phenyl, substituted or unsubstituted benzyl, wherein the substitution refers to being substituted by one or more of C1-C6 alkyl, C1-C6 alkoxy, amino, hydroxyl, halogen, nitro, and trifluoromethyl.

[0023] R3 is selected from hydrogen, deuterium, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, 1,1-dimethyl, 1,5-dimethylhexyl, 1,1-diethanolyl, propenyl, allyl, methoxy, ethoxy, propoxy, butoxy, substituted or unsubstituted phenyl, substituted or unsubstituted benzyl, wherein the substitution refers to being substituted by one or more of C1-C6 alkyl, C1-C6 alkoxy, amino, hydroxyl, halogen, nitro, and trifluoromethyl.

[0024] Further, R1 is selected from methyl 2-propanoate, ethyl acetate, methyl 2-(3-methyl)butanoate, methyl 2-(3-methyl)valerate, methyl 2-(4-methyl)valerate, methyl 2-(3-hydroxy)propanoate, methyl 2-(4-methylthio)butanoate, methyl 2-(4-methoxycarbonyl)butanoate, methyl 2-(3-phenyl)propanoate, and methyl 2-(3-(3-indolyl))propanoate.

[0025] R2 is selected from 3-pyridinemethyl, 2-pyridinemethyl, 4-fluorobenzyl, 4-chlorobenzyl, benzyl, 2-furanmethyl, n-butyl, isopropyl, n-propyl, ethyl;

[0026] R3 is selected from isopropyl, 2-(1-methylthio)ethyl, benzyl, and 3-indolemethyl.

[0027] Furthermore, the benzotriazole compound is selected from the following compounds:

[0028]

[0029] This invention provides a method for preparing the benzotriazole compounds, the synthetic route of which is as follows:

[0030]

[0031] The present invention provides a composition containing the benzotriazole compound or its stereoisomer or its salt or its solvate.

[0032] The present invention provides the use of the composition in the preparation of fungicides, insecticides or herbicides.

[0033] Furthermore, the formulation of the fungicide, insecticide, or herbicide is selected from emulsifiable concentrate (EC), powder (DP), wettable powder (WP), granules (GR), aqueous solution (AS), suspension concentrate (SC), ultra-low volume spray (MLV), soluble powder (SP), microcapsule (MC), fumigant (FU), emulsion (EW), or water-dispersible granules (WG).

[0034] This invention provides the application of the composition in the prevention and control of agricultural pests and diseases, wherein the agricultural pests and diseases are bacterial, fungal, or viral plant diseases. Preferably, the agricultural pests and diseases are plant leaf blight and plant canker; more preferably, the agricultural pests and diseases are rice bacterial leaf blight, cucumber bacterial leaf blight, konjac bacterial leaf blight, citrus canker, tobacco bacterial wilt, grape canker, tomato canker, kiwifruit canker, apple canker, cucumber gray mold, pepper wilt, rapeseed sclerotinia stem rot, wheat scab, potato late blight, blueberry root rot, grape cysticercosis, dragon fruit anthracnose, rice sheath blight, and tobacco mosaic virus; most preferably, the agricultural pests and diseases are tobacco mosaic virus.

[0035] This invention provides the application of the composition in protecting plants from agricultural pests and diseases, wherein the agricultural pests and diseases are bacterial, fungal, or viral diseases of plants. Preferably, the agricultural pests and diseases are plant leaf blight and plant canker; more preferably, the agricultural pests and diseases are rice bacterial leaf blight, cucumber bacterial leaf blight, konjac bacterial leaf blight, citrus canker, tobacco bacterial wilt, grape canker, tomato canker, kiwifruit canker, apple canker, cucumber gray mold, pepper wilt, rapeseed sclerotinia stem rot, wheat scab, potato late blight, blueberry root rot, grape cysticercosis, dragon fruit anthracnose, rice sheath blight, and tobacco mosaic virus; most preferably, the agricultural pests and diseases are tobacco mosaic virus.

[0036] The present invention also provides a method for preventing and controlling agricultural pests and diseases, wherein the benzotriazole compound or composition is applied to the harmful substance or its living environment.

[0037] The present invention discloses the following technical effects:

[0038] This invention is based on benzotriazole and synthesizes a series of benzotriazole compounds containing amide-substituted amino acid esters or amino acid structures. It has been found that these compounds have good inhibitory effects on plant pathogenic viruses, such as tobacco mosaic virus, providing an important scientific basis for the research and development of new pesticides. Detailed Implementation

[0039] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0040] It should be understood that the terminology used in this invention is merely for describing particular embodiments and is not intended to limit the invention. Furthermore, with respect to numerical ranges in this invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Any stated value or intermediate value within a stated range, as well as each smaller range between any other stated value or intermediate value within said range, is also included in this invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0041] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. While only preferred methods and materials have been described herein, any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this invention. All references to this specification are incorporated by way of citation to disclose and describe methods and / or materials associated with those references. In the event of any conflict with any incorporated reference, the content of this specification shall prevail.

[0042] Various modifications and variations can be made to the specific embodiments described in this specification without departing from the scope or spirit of the invention, as will be apparent to those skilled in the art. Other embodiments derived from this specification will also be readily apparent to those skilled in the art. This specification and embodiments are merely exemplary.

[0043] The terms “include,” “including,” “have,” “contain,” etc., used in this article are all open-ended terms, meaning that they include but are not limited to.

[0044] In this invention, the term "alkyl" refers to both branched and straight-chain saturated hydrocarbon groups having a specific number of carbon atoms. For example, "C 1-10 Alkyl (or alkylene) refers to C1, C2, C3, C4, C5, C6, C7, C8, C9, and C10 alkyl groups. Additionally, for example, "C 1-6"Alkyl" means an alkyl group having 1 to 6 carbon atoms. Alkyl groups can be unsubstituted or substituted, such that one or more of their hydrogen atoms are replaced by other chemical groups. Examples of alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (such as n-propyl and isopropyl), butyl (such as n-butyl, isobutyl, tert-butyl), pentyl (such as n-pentyl, isopentyl, neopentyl) and the like.

[0045] In this invention, the term "alkenyl" refers to hydrocarbons that include both straight-chain and branched structures and have one or more carbon-carbon double bonds present at any stable point in the chain. For example, "C..." 2-6 The term "alkenyl" (or "alkenylidene") aims to include C2, C3, C4, C5, and C6 alkenyl groups. Examples of alkenyl groups include, but are not limited to, vinyl, 1-propenyl, 2-propenyl, 2-butenyl, 3-butenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, 5-hexenyl, 2-methyl-2-propenyl, 4-methyl-3-pentenyl, and their analogues.

[0046] In this invention, the term "alkynyl" refers to hydrocarbons that include both straight-chain and branched structures and have one or more carbon-carbon triple bonds present at any stable point in the chain. For example, "C 2-6 The purpose of "alkynyl" (or ynylene) is to include C2, C3, C4, C5 and C6 alkynyl groups; such as ethynyl, propynyl, butynyl, pentylyl, hexynyl and their analogues.

[0047] In this invention, the term "substitution" refers to the substitution of any one or more hydrogen atoms on a specified atom or group by a selected specified group, provided that the substitution does not exceed the general valence of the specified atom. Unless otherwise specified, substituents are named to the central structure. For example, it can be understood that when (cycloalkyl)alkyl is a possible substituent, the point of connection of the substituent to the central structure is in the alkyl moiety. Cyclic double bonds used herein are double bonds formed between two adjacent ring atoms (e.g., C=C, C=N, or N=N). When referring to substitution, particularly polysubstitution, it means that multiple substituents are substituted at various positions on a specified group, such as dichlorophenyl referring to 1,2-dichlorophenyl, 1,3-dichlorophenyl, 1,4-dichlorophenyl, and 2,4-dichlorophenyl.

[0048] Combinations of substituents and / or variables are permitted only when these combinations yield stable compounds or useful synthetic intermediates. A stable compound or stable structure implies that the compound is sufficiently stable to be isolated from the reaction mixture with useful purity, subsequently formulated to form an effective therapeutic agent. Preferably, the compound currently does not contain N-halogens, S(O)₂H, or S(O)H groups.

[0049] In this invention, the term "aryl" refers to a monocyclic or bicyclic aromatic hydrocarbon group, such as phenyl and naphthyl, having 6 to 12 carbon atoms in the ring portion, each of which may be substituted.

[0050] In this invention, the term "halogen" or "halogen atom" refers to chlorine, bromine, fluorine, and iodine.

[0051] In this invention, the term "haloalkyl" refers to a substituted alkyl group having one or more halogen substituents. For example, "haloalkyl" includes mono, bis, and trifluoromethyl groups; even if the halogen in a haloalkyl group is specifically defined as fluorine, chlorine, bromine, or iodine, it still refers to a substituted alkyl group having one or more fluorine, chlorine, bromine, or iodine substituents.

[0052] In this invention, the term "heteroaryl" refers to substituted and unsubstituted aromatic 5- or 6-membered monocyclic groups, 9- or 10-membered bicyclic groups, and 11- to 14-membered tricyclic groups, having at least one heteroatom (O, S, or N) in at least one ring, wherein the heteroatom-containing ring preferably has 1, 2, or 3 heteroatoms selected from O, S, and N. Each ring of the heteroatom-containing heteroaryl may contain one or two oxygen or sulfur atoms and / or 1 to 4 nitrogen atoms, provided that the total number of heteroatoms in each ring is 4 or less, and each ring has at least one carbon atom. The fused ring completing the bicyclic and tricyclic groups may contain only carbon atoms and may be saturated, partially saturated, or unsaturated. Nitrogen and sulfur atoms may optionally be oxidized, and nitrogen atoms may optionally be quaternized. Bicyclic or tricyclic heteroaryl groups must include at least one fully aromatic ring, and the other fused rings may be aromatic or non-aromatic. The heteroaryl group may be attached to any available nitrogen or carbon atom in any ring. Where valence permits, if the other ring is a cycloalkyl or heterocyclic ring, it may optionally be substituted with =O (oxygen).

[0053] Exemplary monocyclic heteroaryl groups include pyrrole, pyrazolyl, pyrazolinyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, thiadiazolyl, furanyl, thiophenyl, oxadiazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, triazinyl, and their analogues.

[0054] Exemplary bicyclic heteroaryl groups include indolyl, benzothiazolyl, benzodioxazolyl, benzoxazolyl, benzothiophenyl, quinolinyl, tetrahydroisoquinolinyl, isoquinolinyl, benzoimidazolyl, benzofuranyl, indoleazinyl, benzofuranyl, crononeyl, coumarinyl, benzofuranyl, cenolinyl, quinoxalinyl, indazoleyl, pyrrolopyridyl, fluoropyridyl, dihydroisoindolyl, tetrahydroquinolinyl, and their analogues.

[0055] Unless otherwise specified, the compounds of this invention are understood to include both their free state and their salts. The term "salt" means an acidic and / or basic salt formed from an inorganic and / or organic acid and base. Additionally, the term "salt" may include zwitterions (internal salts), such as when a compound of formula I contains a basic segment such as an amine or pyridine or imidazole ring, and an acidic segment such as a carboxylic acid. Pharmaceutically acceptable (i.e., non-toxic, physiologically acceptable) salts are preferred, such as acceptable metal and amine salts, wherein the cation does not significantly contribute to toxicity or the biological activity of the salt. However, other salts may be useful, such as those prepared using separation or purification steps, and are therefore also included within the scope of this invention.

[0056] Preferably, C1-C 10 Alkyl groups refer to methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, and their isomers; C1-C 10 Alkoxy refers to methoxy, ethoxy, propoxy, butoxy, pentoxy, hexoxy, heptoxy, octoxy, nonoxy, decoxy and their isomers; C2-C5 alkenyl refers to vinyl, propenyl, allyl, butenyl, pentenyl and their isomers.

[0057] When referring to substituents as alkenyl, alkynyl, alkyl, halogen, aryl, heteroaryl, alkoxy, cycloalkyl, hydroxyl, amino, mercapto, or phosphinyl, or when these substituents specifically refer to a particular alkenyl, alkynyl, alkyl, halogen, aryl, heteroaryl, alkoxy, cycloalkyl, hydroxyl, amino, mercapto, or phosphinyl group, it refers to one to three of the aforementioned substituents. For example, methylphenyl refers to a phenyl group with one to three methyl-substituted groups.

[0058] Example: Preparation of (4-(2-(1H-benzo[d][1,2,3]triazol-1-yl)acetamido)benzoyl)alanine methyl ester

[0059] An amide-carboxylic acid intermediate containing benzotriazole (0.29 g, 1 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.21 g, 1.1 mmol), 1-hydroxybenzotriazole (0.14 g, 1.1 mmol), and dichloromethane (10 mL) were sequentially added to a 25 mL round-bottom flask. After stirring until the solid was completely dissolved, alanine methyl ester hydrochloride (0.15 g, 1.1 mmol) (pre-dissolved in 3 mL of dichloromethane) was added. The reaction was carried out under ice bath for 8 h, and then the reaction was stopped. The sample was washed with NaHCO3 solution (20 mL × 2) and brine (20 mL × 2), respectively, extracted with dichloromethane, dried over MgSO4, filtered, and concentrated under reduced pressure. Finally, the sample was purified by thin-layer chromatography (CH3OH / CH2Cl2, 1:15) to give a white solid in 65.7% yield.

[0060] The structures and 1H and 1C NMR spectra of the synthesized benzotriazole compounds containing amide-substituted amino acid esters or amino acid structures are shown in Table 1, and their physicochemical properties are shown in Table 2.

[0061] Table 1. 1H NMR, 1C NMR, and high-resolution mass spectrometry data of the compounds.

[0062]

[0063]

[0064]

[0065]

[0066]

[0067]

[0068]

[0069]

[0070]

[0071]

[0072]

[0073]

[0074]

[0075]

[0076]

[0077]

[0078]

[0079]

[0080] Table 2 Physicochemical properties of the target compounds

[0081]

[0082]

[0083] Pharmacological Example 1:

[0084] Application of benzotriazole compounds containing amide-substituted amino acid esters or amino acid structures in antiviral studies, specifically in assays against tobacco mosaic virus.

[0085] The antiviral activity of the compound was determined using the half-leaf spot method. 3 mg of the test compound was accurately weighed into a weighing bottle and dissolved completely in 60 μL of DMSO solvent. A 500 mg / L compound solution was prepared by dissolving the compound in double-distilled water containing 1% (v / v) Tween 20. Separately, 3 mg of ribavirin was dissolved in 60 μL of DMSO solvent and dissolved in double-distilled water containing 1% Tween 20 to prepare a 500 mg / L ribavirin solution.

[0086] The in vivo therapeutic activity of the agent against TMV infection. Uniformly growing heart-leaf tobacco plants were selected, and virus solution (concentration 6 × 10⁻⁶) was first collected using a parallel dipper. -3 (mg / mL) The virus solution was manually inoculated onto the leaf surface (whole leaf) along the veins, using a friction rub, ensuring consistent inoculation on both leaves. The leaves were supported by a flat wooden board underneath. After the virus solution dried, the emery was rinsed off the leaves with running water. After the leaves dried, the left half of the leaf was treated with the solution, and the right half was treated with sterilized water as a control. Each treatment consisted of 3 plants, each with 3-4 leaves. The plants were then placed in a light incubator with humidity control at 23℃ and 10000 Lux of light. The number of necrotic spots was observed and recorded after 2-4 days. Each treatment was repeated 3 times, and the inhibition rate was calculated.

[0087] The in vivo protective activity of the agent against TMV infection was investigated. Uniformly growing heart-leaf tobacco plants were selected, and the agent was gently applied to the left half of the leaf with a brush. The right half of the leaf was treated with sterile water as a control. Virus was inoculated 24 hours later. Virus extract (concentration 6 x 10⁻⁶) was collected using a brush. -3 (mg / mL) The virus solution was manually inoculated onto the leaf surface (whole leaf) along the veins, using a friction rubbing motion. The inoculation intensity on both leaves should be kept as consistent as possible. The leaves were supported under a flat wooden board. After the virus solution dried, the emery was rinsed off the leaves with running water. Three plants were treated with each agent, each with 3-4 leaves. The plants were then placed in a light incubator with humidity control at 23℃ and 10000 Lux light. After 2-4 days, the number of necrotic spots was observed and recorded. Each agent was repeated three times as described above, and the inhibition rate was calculated. Y(%) = (RL) / R × 100%

[0088] Wherein: Y represents the inhibition rate of the compound against tobacco mosaic virus; R represents the number of necrotic spots in the control group (right half of the leaf); and L represents the number of necrotic spots in the treatment group (left half of the leaf). The embodiments of this invention are provided to illustrate the technical solution of this invention, but the content of the embodiments is not limited thereto. The experimental results are shown in Table 3.

[0089] Table 3. Inhibitory activity of benzotriazole compounds with amide-substituted amino acid esters or amino acid structures against tobacco mosaic virus at a concentration of 500 mg / mL.

[0090]

[0091]

[0092]

[0093]

[0094] Table 3 shows that the target compound exhibited slightly higher antiviral activity against tobacco mosaic virus (TMV) at 500 mg / mL than the control agent, ribavirin. Anti-TMV activity results indicated that the target compound with amino acid ester substitution generally showed higher antiviral inactivation activity than aryl amines and aliphatic amine substituents, as listed below: 3f (70.1%) > 3a (52.6%) > 3h (38.1%). Furthermore, in terms of protective activity, the ortho-position of the compound showed higher primary anti-TMV activity than the para-position, such as 3f (58.5%) > 1g (41.7%), 3c (62.7%) > 2e (41.3%), and 3b (57.4%) > 2b (53.6%). For primary amine substituents containing aromatic rings, introducing electron-withdrawing groups such as fluorine (3e) or chlorine (3d) onto the benzene ring significantly reduced the antiviral protective ability of the target compound. To investigate the effect of the aromatic ring in the R group on biological activity, groups containing aliphatic rings were introduced into the molecular structure. However, bioassay results showed that the target compound did not improve the anti-TMV activity, indicating that the aliphatic amine-substituted groups did not contribute to the activity compared to the aromatic ring.

[0095] Pharmacological Example 2:

[0096] EC 50 Median effective concentration (EC50) is an important indicator for evaluating the sensitivity of plant pathogen viruses to compounds, and it is also a crucial parameter for setting the compound concentration when studying the mechanism of action of target compounds. In concentration gradient experiments, five appropriate concentrations were set using the two-fold dilution method. Finally, the inhibition rate of the agent against the plant pathogen virus and the agent concentration were converted into logarithmic values, and the virulence curve was obtained through regression analysis using SPSS software to calculate the EC50. 50 .

[0097] The antiviral activity of the compounds was determined using the half-leaf spot method. 5 mg of the test compound and the control agent ribavirin were accurately weighed into separate weighing bottles, and 100 μL of DMSO solvent was added to dissolve them completely. Compound solutions of 500, 400, 300, 200, and 100 mg / L were prepared using double-distilled water containing 1% Tween 20.

[0098] The in vivo therapeutic activity of the agent against TMV infection was investigated. Uniformly growing *Nicotiana macrophylla* plants were selected. Virus solution (concentration 6 × 10⁻³ mg / mL) was first applied using a dipper to the entire leaf surface along its veins, then manually rubbed onto leaves sprinkled with carborundum (emery powder). The inoculation intensity on both leaves was kept as consistent as possible, and the leaves were supported by a flat wooden board underneath. After the virus solution dried, the carborundum was rinsed off the leaves with running water. After the leaves dried, the agent was applied to the left half of the leaf, and sterilized water was applied to the right half as a control. Three plants were treated with each agent, each with 3-4 leaves. The plants were then placed in a light-controlled incubator with humidity at 23℃ and 10000 Lux of light. The number of necrotic spots was observed and recorded after 2-4 days. Each agent was repeated three times, and the inhibition rate was calculated.

[0099] The in vivo protective activity of the agent against TMV infection was investigated. Uniformly growing *Nicotiana siceraria* plants were selected. The agent was gently applied to the left half of the leaf with a brush, while the right half was treated with sterile water as a control. The leaves were inoculated with the virus 24 hours later. Using a brush, the virus extract (concentration 6 x 10⁻³ mg / mL) was manually rubbed onto the leaf surface (whole leaf) along its veins onto the leaf sprinkled with carborundum. The inoculation intensity on both leaves was kept as consistent as possible, and the leaves were supported under a flat wooden board. After the virus extract dried, the carborundum was rinsed off the leaves with running water. Three plants were treated with each agent, each with 3-4 leaves. The plants were then placed in a light incubator with humidity control at 23℃ and 10000 Lux light. The number of necrotic spots was observed and recorded after 2-4 days. Each agent was replicated three times, and the inhibition rate was calculated. Y(%) = (RL) / R × 100%

[0100] Wherein: Y represents the inhibition rate of the compound against tobacco mosaic virus; R represents the number of necrotic spots in the control group (right half of the leaf); and L represents the number of necrotic spots in the treatment group (left half of the leaf). The embodiments of this invention are provided to illustrate the technical solution of this invention, but the content of the embodiments is not limited thereto. The experimental results are shown in Table 4.

[0101] Table 4. Benzotriazole compounds with amide-substituted amino acid esters or amino acid structures exhibiting activity against the EC50 of tobacco mosaic virus. 50

[0102]

[0103] As shown in Table 4, the antiviral activity of the target compounds changed differently after conversion from ester to acid. When the methylene group on the amino acid was linked to a substituent containing an aromatic ring, the antiviral activity was significantly enhanced, especially compound 4d (R = 3-indolemethyl, 85.1% at 500 μg / mL, 38.4% at 100 μg / mL), which exhibited the highest activity and was significantly higher than that of the commercially available drug ribavirin (51.6% at 500 μg / mL, 18.3% at 100 μg / mL). Given that the target compounds showed a clear trend of enhanced antiviral inactivation activity after hydrolysis from ester to acid, to further accurately evaluate the relationship between the carboxyl group and biological activity, EC50 tests were performed on compounds 1i, 1j, 4a, 4b, 4c, and 4d to assess their antiviral inactivation activity. As shown in Table 3, the substituents on the molecular backbone affect the overall antiviral ability, with corresponding EC50 values ​​as follows: 4d (157.6 μg / mL) > 4c (204.5 μg / mL) > 1j (304.7 μg / mL) > 1i (313.8 μg / mL). All of these values ​​demonstrate superior efficacy compared to ribavirin (EC50 = 442.1 μg / mL), indicating that introducing amino acids with aromatic ring substitutions into the target molecule can promote inactivation against TMV. This can be used to prepare pesticides against plant pathogenic viruses.

[0104] The embodiments described above are merely preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Various modifications and improvements made by those skilled in the art to the technical solutions of the present invention without departing from the spirit of the present invention should fall within the protection scope defined by the claims of the present invention.

Claims

1. A benzotriazole compound, characterized in that, Selected from the following compounds: , , , , , , , , , , , , , , , , , , , , , or .

2. A composition, characterized in that, Contains the benzotriazole compound of claim 1 or a salt thereof.

3. The use of the composition of claim 2 in the preparation of a drug for inhibiting tobacco mosaic virus.

4. The use of the composition of claim 2 in the prevention and control of tobacco mosaic virus.

5. The use of the composition of claim 2 in protecting plants from tobacco mosaic virus.

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