Purine nucleoside compound containing cinnamic acid structure as well as preparation method and application of purine nucleoside compound

Abstract

The invention discloses a purine nucleoside compound containing a cinnamic acid structure as well as a preparation method and application of the purine nucleoside compound, and a series of purine nucleoside compounds containing the cinnamic acid structure are synthesized by taking a purine nucleoside compound as a basis and introducing a piperazine-containing cinnamic acid fragment into a system. The compound has a good inhibition effect on plant pathogenic microorganisms, and especially has a good inhibition effect on plant pathogenic viruses such as tobacco mosaic viruses. Wherein the activity data of a part of the compounds is superior to that of a commercial drug ribavirin, the activity of the compounds is equivalent to that of ningnanmycin, particularly, the compound 14 has excellent treatment activity, and the EC50 of the compound 14 is 274.25 mu g / mL. The compound has good antibacterial activity on Xanthomonas oryzae pv. Oryzae, when the concentration is 50 [mu] g / mL, the inhibitory activities of the compound 7, the compound 20 and the compound 21 on Xanthomonas oryzae pv. Oryzae are 36.2%, 40.3% and 35.6% respectively and are superior to 32.6% of a control agent thiediazole copper, when the concentration is 50 [mu] g / mL or below, the inhibitory effects of the compound 1, the compound 3, the compound 23 and the compound 24 on Xanthomonas citri are 21.3%, 20.4%, 27.8% and 22.6% respectively, and the compound 7, the compound 20 and the compound 21 on Xanthomonas oryzae pv. Oryzae are superior to that of the control agent thiediazole copper. And the results are better than that of a control agent thiediazole copper by 18.6%.

Description

Technical Field

[0001] The present invention belongs to the technical field of medicinal chemistry, and particularly relates to a class of purine nucleoside compounds containing cinnamic acid structure, and their preparation methods and applications. Background Art

[0002] Plant diseases caused by stubborn phytopathogenic viruses have greatly increased the burden on the field ecosystem, severely restricting the yield and quality of crops or horticultural crops. Plant viruses cause economic losses of more than $100 billion to global agriculture every year. Pesticide application remains the fastest and most effective means of preventing and controlling crop damage caused by plant pathogens, and is also a practical means of reducing labor costs. Currently, the widely used commercial antiviral agents ribavirin and ningnanmycin are only marginally effective in the field control of tobacco mosaic virus (TMV). In addition, due to increasingly prominent problems such as drug resistance, cross-resistance, agricultural residues, and environmental safety, modern agriculture faces difficult challenges. Therefore, it is imperative to explore an environmentally friendly active pesticide skeleton with a targeted effect. Bioactive plant-derived natural products are important resources for the creation and research of new pesticides.

[0003] Purine nucleosides are important endogenous substances in organisms and an important part of genetic materials. Their derivatives have broad-spectrum biological activities, such as anti-tumor, antibacterial, anti-inflammatory, insecticidal, antiviral, etc., especially showing excellent performance in antiviral. It is estimated that so far, there are about 100 drugs developed based on purine in medicine, and more than half of them are antiviral drugs. The mechanism of action of such drugs is mainly to inhibit the synthesis of viral nucleic acids and increase the frequency of lethal mutations in the viral genome to achieve the antiviral effect. In addition, cinnamic acid widely exists in plants such as cinnamon and laurel, and has a wide range of biological activities. However, the research on the anti-plant virus of the two is still in its infancy and has good application prospects.

[0004] The research progress of the biological activities of purine nucleoside compounds is as follows:

[0005] In 2018, Sekgota et al. synthesized a series of new 2 - aminoadenine and - guanine 2′ - deoxynucleoside analogues through glycosylation, hydrogenation, and SNAr reactions. The C - 4′ position of the sugar moiety was substituted with methyl, fluoromethyl, vinyl, ethyl, or azide groups, and their antiviral activities against HIV - 1 and HBV were determined. The results showed significant to potent anti - HIV and HBV activities, and also exhibited low cytotoxicity. In 2018, Hulpia et al. constructed a rich nucleoside library around the 3′ - C - ethynyl - D - ribofuranose sugar scaffold. A series of novel purine nucleoside derivatives were obtained by coupling the sugar scaffold with various modified purine nucleobases, and the ability of the resulting compounds to inhibit tumor cell proliferation was further evaluated. It was found that compound 21 showed excellent antitumor properties against L1210, CEM, and HeLa cells, with IC 50 values of 0.014 μM, 0.012 μM, and 0.051 μM, respectively. In addition, BLI analysis showed that it could inhibit tumor growth and reduce the formation of lung metastases. These results indicate that the recombination of known sugars and nucleobases is an effective design strategy for discovering new antitumor agents. In 2018, Zhou et al. synthesized a series of novel chalcone derivatives containing purine and benzenesulfonamide structures and tested their antiviral activities against TMV. The results showed that some derivatives exhibited anti - TMV activity. In 2019, He et al. designed and synthesized a series of novel purine nucleoside derivatives containing sulfonamide structures. Through anti - CMV activity testing, the results showed that the most excellent compound had excellent inactivation activity against CMV, with an EC 50 value of 48.8 μg / mL, superior to the control drugs ningnanmycin (EC 50 value of 84.7 μg / mL) and ribavirin (EC 50 value of 150.4 μg / mL). In 2024, Deng et al. synthesized a series of quinazolinone derivatives containing purine structures. After activity testing, the results showed that some compounds exhibited higher antiviral activities against tobacco mosaic virus (TMV) than ribavirin. Among them, the most excellent compound showed significant therapeutic activity against TMV, with an EC 50 value of 162.3 μg / mL, superior to ribavirin (314.4 μg / mL).

[0006] However, the research on purine nucleosides and their derivatives in the field of anti - plant viruses is in its initial stage. Therefore, it is of crucial significance to provide a purine nucleoside and its derivatives with high antiviral activity. Summary of the Invention

[0007] To solve the above - mentioned technical problems, the present invention provides a class of purine nucleoside compounds containing cinnamic acid structures, their preparation methods, and applications.

[0008] To achieve the above object, the present invention provides a class of purine nucleoside compounds containing a cinnamic acid structure, which have the structure shown in the general formula (I):

[0009]

[0010] Wherein: R is selected from one or more of substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl or heterocycle; more preferably, R is one or more of substituted or unsubstituted C6-C15 aryl, substituted or unsubstituted C6-C10 heteroaryl; R is independently 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, phenyl, 2-methoxyphenyl, 3-methoxyphenyl, 4-methoxyphenyl, 2,3-dimethoxyphenyl, 3,4-dimethoxyphenyl, 3,4,5-trimethoxyphenyl, 2-hydroxyphenyl, 4-hydroxyphenyl, 4-methylphenyl, 4-diethylaminophenyl, benzo(1,3)dioxolyl, 4-fluorophenyl, 4-trifluorophenyl, 2-chlorophenyl, 4-chlorophenyl, 2,4-dichlorophenyl, 3,4-dichlorophenyl, 3-bromophenyl, 4-bromophenyl, 4-cyanophenyl, 2-nitrophenyl, 3-nitrophenyl, 3-nitrophenyl, 4-nitrophenyl, 3-pyridyl, 2-furyl, 4-methylthiazolyl, 2-thienyl. Wherein, the substitution means being substituted by one or more of C1-C6 alkyl, C1-C6 alkoxy, amino, hydroxyl, halogen, nitro and trifluoromethyl.

[0011] Further, the class of purine nucleoside compounds containing cinnamic acid are selected from the following compounds: R is independently selected from phenyl, 2-methoxyphenyl, 3-methoxyphenyl, 4-methoxyphenyl, 2,3-dimethoxyphenyl, 3,4-dimethoxyphenyl, 3,4,5-trimethoxyphenyl, 2-hydroxyphenyl, 4-hydroxyphenyl, 4-methylphenyl, 4-diethylaminophenyl, benzo(1,3)dioxolyl, 4-fluorophenyl, 4-trifluorophenyl, 2-chlorophenyl, 4-chlorophenyl, 2,4-dichlorophenyl, 3,4-dichlorophenyl, 3-bromophenyl, 4-bromophenyl, 4-cyanophenyl, 2-nitrophenyl, 3-nitrophenyl, 4-nitrophenyl, 3-pyridyl, 2-furyl, 4-methylthiazolyl, 2-thienyl.

[0012] A preparation method of the purine nucleoside compound containing a cinnamic acid structure, comprising the following steps:

[0013] Preparation method of the purine nucleoside compound containing a cinnamic acid structure:

[0014]

[0015] R is selected from hydrogen, deuterium, optionally substituted or unsubstituted alkyl, optionally substituted or unsubstituted alkenyl, optionally substituted or unsubstituted alkynyl, optionally substituted or unsubstituted alkoxy, optionally substituted or unsubstituted cycloalkyl, optionally substituted or unsubstituted aryl, or optionally substituted or unsubstituted heteroaryl.

[0016] A pharmaceutical composition comprising the purine nucleoside compound containing a cinnamic acid structure as described above, or a stereoisomer thereof, or a salt thereof, or a solvate thereof, and an agriculturally acceptable adjuvant or an antiviral agent, an insecticide or a herbicide.

[0017] Furthermore, the dosage form of the above pharmaceutical composition is selected from emulsifiable concentrate (EC), dust (DP), wettable powder (WP), granule (GR), aqueous solution (AS), suspension concentrate (SC), ultra-low volume spray (ULV), soluble powder (SP), microcapsule (MC), smoke agent (FU), emulsion in water (EW), or water dispersible granule (WG).

[0018] Use of the purine nucleoside compound containing a cinnamic acid structure as described above or the above pharmaceutical composition in controlling agricultural pests and diseases, wherein the agricultural pests and diseases are plant viral diseases, plant bacterial diseases, plant fungal diseases, or plant insect pests.

[0019] A method for controlling agricultural pests and diseases, which comprises allowing the purine nucleoside compound containing cinnamic acid as described above or the above pharmaceutical composition to act on the harmful substances or their living environment.

[0020] A method for protecting plants from agricultural pests and diseases, which comprises bringing the purine nucleoside compound containing a cinnamic acid structure as described above or the above pharmaceutical composition into contact with the plants.

[0021] Furthermore, the agricultural pests and diseases are plant bacterial diseases, plant fungal diseases, or plant viral diseases; further, the agricultural pests and diseases are plant viral diseases; still further, the agricultural pests and diseases are viral diseases such as tobacco mosaic virus, cucumber mosaic virus, potato virus Y, etc.; bacterial diseases such as rice bacterial blight, citrus canker, tobacco bacterial wilt, kiwifruit canker, rice bacterial leaf streak, etc.; fungal diseases such as cucumber gray mold, pepper fusarium wilt, rape sclerotinia, wheat scab, potato late blight, blueberry root rot, botryosphaeria dothidea, pitaya anthracnose, rice sheath blight, etc.; insect diseases such as green worms, caterpillars, aphids, scale insects, whiteflies, etc.; nematode diseases such as root-knot nematodes, dagger nematodes, stem nematodes, etc.; mite diseases such as cotton spider mites, citrus red spider mites, etc., and still further, the agricultural pests and diseases are tobacco mosaic virus diseases.

[0022] Compared with the prior art, the present invention has the following advantages and technical effects:

[0023] To provide a new class of natural lead compounds with high antiviral activity, the present invention is based on purine nucleoside compounds, synthesizes a series of purine nucleoside derivatives containing cinnamic acid structure, and examines their biological activities against plant pathogenic viruses. It is found that this class of compounds has good inhibitory effects on plant pathogenic viruses and has good inhibitory effects against pathogenic viruses such as tobacco mosaic virus. In in vivo experiments, the target compounds showed good inhibitory activities against the plant pathogenic virus TMV. The EC50 values of some compounds such as 13, 14, 16, and 17 were 334.78, 274.25, 327, 382, 333.42, and 255.6 μg / mL, respectively. The tested activity data were all better than that of the commercial drug ribavirin (783.54 μg / mL), and the activity of the compounds was comparable to that of ningnanmycin (283.26 μg / mL). From the above, it can be seen that purine nucleoside compounds containing cinnamic acid have good antiviral activities. In particular, compound 14 has excellent therapeutic activity with an EC50 of 274.25 μg / mL. Some compounds showed good antibacterial activities against Xanthomonas oryzae pv. oryzae (Xoo). At a concentration of 50 μg / mL, the inhibitory activities of compounds 7, 20, and 21 against Xoo were 36.2%, 40.3%, and 35.6%, respectively, which were better than that of the control agent thiodiazole copper (32.6%). At a concentration below 50 μg / mL, the inhibitory effects of compounds 1, 3, 23, and 24 against Pseudomonas syringae pv. actinidiae (Psa) were 21.3%, 20.4%, 27.8%, and 22.6%, respectively, all of which were better than that of the control agent thiodiazole copper (18.6%). From the above, it can be seen that purine nucleoside compounds containing cinnamic acid structure have good antibacterial activities. Detailed Description of the Invention

[0024] The 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, characteristics, and embodiments of the present invention.

[0025] It should be understood that the terms used in the present invention are only for describing specific embodiments and are not used to limit the present invention. Additionally, for the numerical ranges in the present invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Each intermediate value within any stated value or stated range, as well as each smaller range between any other stated value or intermediate value within the stated range, is also included in the present invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0026] Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. Although this invention has been described only in terms of preferred methods and materials, any methods and materials similar or equivalent to those described herein can also be used in the practice or testing of this invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials related to those documents. In case of conflict with any incorporated document, the content of this specification shall prevail.

[0027] Without departing from the scope or spirit of the present invention, various modifications and variations to the specific embodiments of the description of the present invention will be apparent to those skilled in the art. Other embodiments derived from the description of the present invention will be apparent to those skilled in the art. The description of the present invention and the examples are merely exemplary.

[0028] Regarding the use of "comprising", "including", "having", "containing", etc. herein, they are all open-ended terms, meaning including but not limited to.

[0029] The term "alkyl" as used herein includes both branched and straight-chain saturated hydrocarbon groups having a specific number of carbon atoms. For example, "C1-10 alkyl" (or alkylene) is intended to mean C1, C2, C3, C4, C5, C6, C7, C8, C9, and C10 alkyl. Additionally, for example, "C1-6 alkyl" represents an alkyl group having 1 to 6 carbon atoms. The alkyl group can be unsubstituted or substituted such that one or more of its 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.

[0030] The term "alkenyl" as used herein includes both straight-chain or branched-chain hydrocarbons having one or more carbon-carbon double bonds at any stable point in the chain. For example, "C2-6 alkenyl" (or alkenylene) is intended to include C2, C3, C4, C5, and C6 alkenyl. 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 the like.

[0031] The term "alkynyl" as used herein includes both straight-chain or branched-chain hydrocarbons having one or more carbon-carbon triple bonds at any stable point in the chain. For example, "C2-6 alkynyl" (or alkynylene) is intended to include C2, C3, C4, C5, and C6 alkynyl; such as ethynyl, propynyl, butynyl, pentynyl, hexynyl, and the like.

[0032] The term "substituted" as used herein means that any one or more hydrogen atoms on a specified atom or group are replaced by a selected specified group, provided that the general valence of the specified atom is not exceeded. Unless otherwise specified, substituents are named to the central structure. For example, it is understood that when (cycloalkyl)alkyl is a possible substituent, the point of attachment of the substituent to the central structure is in the alkyl moiety. The cyclic double bonds used herein are double bonds formed between two adjacent ring atoms (such as C═C, C═N or N═N). When substitution is mentioned, especially poly-substitution, it means that multiple substituents replace at various positions on the specified group. For example, dichlorophenyl means 1,2-dichlorophenyl, 1,3-dichlorophenyl, 1,4-dichlorophenyl and 2,4-dichlorophenyl. Combinations of substituents and / or variables are permitted only if these combinations result in stable compounds or useful synthetic intermediates. A stable compound or stable structure implies that the compound is sufficiently stable when separated from the reaction mixture in useful purity and subsequently formulated to form an effective therapeutic agent. Preferably, the compounds described herein do not contain N-halogen, S(O)2H or S(O)H groups.

[0033] The term "aryl" as used herein means a monocyclic or bicyclic aromatic hydrocarbon group having 6 to 12 carbon atoms in the ring portion, such as phenyl and naphthyl, each of which may be substituted.

[0034] The term "halogen" or "halogen atom" as used herein refers to chlorine, bromine, fluorine and iodine. The term "haloalkyl" refers to a substituted alkyl group having one or more halogen substituents. For example, "haloalkyl" includes mono-, di- and trifluoromethyl; even if the halogenation in the haloalkyl is specified as fluorine, chlorine, bromine, iodine, it also refers to a substituted alkyl group having one or more fluorine, chlorine, bromine, iodine substituents. 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, and the heteroatom-containing ring preferably has 1, 2 or 3 heteroatoms selected from O, S and N. Each ring of the heteroaryl containing heteroatoms may contain one or two oxygen or sulfur atoms and / or from 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 rings completing the bicyclic and tricyclic groups may contain only carbon atoms and may be saturated, partially saturated or unsaturated. The nitrogen and sulfur atoms may be optionally oxidized and the nitrogen atoms may be optionally quaternized. The bicyclic or tricyclic heteroaryl must include at least one fully aromatic ring, and the other fused rings may be aromatic or non-aromatic. The heteroaryl may be attached to any available nitrogen or carbon atom of any ring. When the valence allows, if the other ring is a cycloalkyl or heterocycle, it is additionally optionally substituted with ═O (oxygen). Exemplary monocyclic heteroaryls include pyrrolyl, pyrazolyl, pyrazolinyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, thiadiazolyl, furyl, thienyl, oxadiazolyl, styryl, pyrazinyl, pyrimidinyl, pyridazinyl, triazinyl and the like. Exemplary bicyclic heteroaryls include indolyl, benzothiazolyl, benzodioxolyl, benzoxazolyl, benzothienyl, quinolinyl, tetrahydroisoquinolinyl, isoquinolinyl, benzimidazolyl, benzofuranyl, indolizinyl, benzofuranyl, chromonyl, oxime ether group, benzofuranyl, cinnolinyl, quinoxalinyl, indazolyl, pyrrolostyryl, fluorostyryl, dihydroisoindolyl, tetrahydroquinolinyl and the like.

[0035] Unless otherwise specified, the compounds of the present invention are understood to include the free form and their salts. The term "salt" refers to acid addition and / or base salts formed with inorganic and / or organic acids and bases. Additionally, the term "salt" may include zwitterions (inner salts), such as when the compound of formula I contains a basic moiety such as an amine or styryl or imidazole ring, and an acidic moiety such as a carboxylic acid. Pharmaceutically acceptable (i.e., non-toxic, physiologically acceptable) salts are preferred, such as acceptable metal and amine salts, where the cation does not significantly contribute to the toxicity or biological activity of the salt. However, other salts may be useful, such as in separation or purification steps during preparation, and are thus also included within the scope of the present invention. Preferably, C1-C10 alkyl refers to methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl and their isomers; C1-C10 alkoxy refers to methoxy, ethoxy, propoxy, butoxy, pentyloxy, hexyloxy, heptyloxy, octyloxy, nonyloxy, decyloxy and their isomers; C2-C5 alkenyl refers to vinyl, propenyl, allyl, butenyl, pentenyl and their isomers. When referring to a substituent being alkenyl, alkynyl, alkyl, halogen, aryl, heteroaryl, alkoxy, cycloalkyl, hydroxy, amino, mercapto, phosphino, or when these substituents are specifically a particular alkenyl, alkynyl, alkyl, halogen, aryl, heteroaryl, alkoxy, cycloalkyl, hydroxy, amino, mercapto, phosphino, it refers to one to three of the above substituents. For example, methylphenyl refers to a phenyl substituted with one to three methyl groups.

[0036] All raw materials and solvents used in the examples of the present invention are commercially available products.

[0037] Example 1

[0038] Synthetic method of the target compound (E)-1-(4-(9-((3S,4R,5S)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-9H-purin-6-yl)piperazin-1-yl)-3-phenylprop-2-en-1-one (1):

[0039] First, weigh 500 mg of benzaldehyde and add it to 8 mL of pyridine solution. Then add 413 mg of malonic acid and stir well. Subsequently, pipette 90 μL of piperidine solution into the above solution and reflux it at 85 °C for 4 hours. After the reaction is completed, wait for the reaction solution to cool, then add hydrochloric acid dropwise to the reaction solution to adjust the pH to 2 - 3. At this time, a solid precipitates. Filter the solid by suction, and finally recrystallize it with ethanol to obtain a white solid, which is intermediate a1. Weigh 3 g of piperazine and dissolve it in 40 mL of ethanol solution. Then weigh 2 g of 6-chloropurine riboside and add it to the above solution, and stir at room temperature for 10 minutes. Finally, pipette 970 μL of triethylamine solution into the above solution and reflux it at 80 °C for 14 - 16 hours. After the reaction is completed, a white solid precipitates. Remove the filtrate by suction filtration to obtain a white solid, which is intermediate b. Weigh 300 mg of trans-cinnamic acid (intermediate a1) and dissolve it in 10 mL of N,N-dimethylformamide (DMF) solution. Then sequentially add 204 mg of 1-hydroxybenzotriazole (HOBT), 288 mg of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI), and 700 mmol of triethylamine solution (Et3N) to the above solution. Stir at room temperature for one hour and then add 508 mg of intermediate b. After stirring at room temperature for 16 - 18 hours, the reaction is completed. Then add an appropriate amount of water. If a solid precipitates, filter the solid by suction. Then dissolve the solid in a mixed solution of dichloromethane and methanol, and finally purify it by column chromatography (DCM:MeOH = 40:1 - 20:1, v:v) to obtain the target compound. If no solid precipitates, extract the organic layer with dichloromethane, and finally purify it by column chromatography (DCM:MeOH = 40:1 - 20:1, v:v) to obtain a yellow solid with a yield of 46.3%.

[0040] The compounds were all synthesized according to the above method.

[0041] The synthesized purine riboside compounds containing cinnamic acid structure and their 1H NMR and 13C NMR data are shown in Table 1, and their physical and chemical properties are shown in Table 2.

[0042] Table 1 1H NMR and 13C NMR data of the compounds

[0043]

[0044]

[0045]

[0046]

[0047]

[0048]

[0049]

[0050]

[0051]

[0052]

[0053]

[0054] Table 2 Physicochemical properties of the target compounds

[0055]

[0056]

[0057] Example 2

[0058] Application of purine nucleoside compounds containing cinnamic acid structure in anti-pathogenic pathogenic virus, for the test method of anti-tobacco mosaic virus (TMV).

[0059] The anti-plant virus activity of the agent was determined by the half-leaf spot method. Accurately weigh 3 mg of the test compound in a weighing bottle, add 60 μL of solvent DMSO to dissolve it completely. Use secondary distilled water containing 1 wt% Tween 20 to prepare a 500 mg / L compound solution. Another 250 μL of 2 wt% ribavirin aqueous solution was added with 60 μL of solvent DMSO and 10 mL of secondary distilled water containing 1 wt% Tween 20 to prepare a 500 mg / L ribavirin solution.

[0060] In vivo therapeutic activity of the agent against TMV infection. Select Nicotiana glutinosa with consistent growth. First, dip a row of brushes into the virus solution (concentration: 6×10 -3 mg / mL), and manually rub and inoculate it on the leaf surface (whole leaf) along the vein direction on the leaf with carborundum sprinkled on it. Try to keep the inoculation force of the left and right leaves as consistent as possible, and support the leaves below with a flat wooden board. After the virus solution dries, wash off the carborundum on the leaf surface with running water. After the leaves dry, apply the agent on the left half leaf and apply sterilized water on the right half leaf as a control. Each agent treatment has 3 plants, and each plant has 3 - 4 leaves. Then place the plants in a light incubator for moisturizing culture, control the temperature at 23 °C, light at 10000 LuX, observe and record the number of necrotic spots after 2 - 4 days. Repeat each agent 3 times according to the above method, and calculate the inhibition rate (Y). The calculation formula is as follows:

[0061] Y(%) = (R - L) / R × 100%

[0062] Wherein: Y is the inhibition rate of the compound against tobacco mosaic virus; R is the number of necrotic spots on the control group (right half leaf); L is the number of necrotic spots on the treatment group (left half leaf).

[0063] Examples of the present invention are used to illustrate the technical solutions of the present invention, but the content of the examples is not limited thereto. The experimental results are shown in Table 3.

[0064] Table 3 Activity test results of purine nucleoside compounds containing cinnamic acid structure against plant pathogenic virus TMV at a concentration of 500 mg / L -1 The concentration was tested for the activity against plant pathogenic virus TMV

[0065]

[0066]

[0067] * The values are the average ± standard deviation of three replicates.

[0068] Example 3

[0069] EC 50 is an important indicator for evaluating the sensitivity of plant pathogenic viruses to compounds, and is also an important parameter for setting the compound concentration when studying the action mechanism of target compounds. In the concentration gradient experiment, five appropriate concentrations were set by the two-fold dilution method. Finally, the inhibition rate of the medicament against plant pathogenic virus and the medicament concentration were converted into logarithmic values, and the virulence curve was obtained by regression analysis using SPSS software, and EC 50 .

[0070] The anti-plant virus activity of the compound was determined by the half-leaf spot method. Accurately weigh 5 mg of the test compound into a weighing bottle, add 100 μl of solvent DMSO to dissolve it completely. The compound solutions with concentrations of 500, 250, 125, 62.5, and 31.25 mg / L were prepared respectively with secondary distilled water containing 1 wt% Tween 20. Another 250 μl of 2% ribavirin aqueous solution was taken, 60 μl of solvent DMSO and 10 mL of secondary distilled water containing 1 wt% Tween 20 were added to prepare a 500 mg / L ribavirin solution.

[0071] The in vivo therapeutic activity of the medicament against TMV infection. Select heartleaf tobacco with consistent growth, first dip the virus solution (concentration of 6×10 -3(mg / mL). Manually inoculate it on the leaf surface (whole leaf) along the direction of its lateral veins onto the leaf blades sprinkled with carborundum. Try to keep the inoculation force of the left and right leaves as consistent as possible, and support the lower part of the leaf with a flat wooden board. After the virus solution dries, rinse off the carborundum on the leaf with running water. After the leaf dries, apply the medicament on the left half leaf and apply sterilized water on the right half leaf as a control. Set 3 plants for each medicament treatment, with 3 - 4 leaves per plant. Then place the plants in a light incubator for moisturizing cultivation, control the temperature at 23°C, the light at 10000 LuX, and observe and record the number of necrotic spots generated after 2 - 4 days. Repeat each medicament 3 times according to the above method, and calculate the inhibition rate (Y). The calculation formula is as follows:

[0072] Y (%) = (R - L) / R × 100%

[0073] Where: Y is the inhibition rate of the compound against tobacco mosaic virus; R is the number of necrotic spots in the control group (right half leaf); L is the number of necrotic spots in the treatment group (left half leaf).

[0074] The examples of the present invention are used to illustrate the technical solutions of the present invention, but the content of the examples is not limited thereto. The experimental results are shown in Table 4.

[0075] Table 4 Therapeutic activity EC of purine nucleoside compounds containing cinnamic acid structure against plant pathogenic virus TMV 50 Test

[0076]

[0077] As can be seen from Table 3 and Table 4, in the in vivo test, the target compounds showed good inhibitory activity against the plant pathogenic virus TMV. The EC50 values of some compounds such as 13, 14, 16, and 17 were 334.78, 274.25, 327, 382, 333.42, and 255.6 μg / mL respectively. All the tested activity data were better than the commercial drug ribavirin (783.54 μg / mL), and the activity of the compounds was comparable to that of ningnanmycin (283.26 μg / mL). From the above, it can be known that purine nucleoside compounds containing cinnamic acid have good antiviral activity. In particular, compound 14 has excellent therapeutic activity, and its EC50 is 274.25 μg / mL.

[0078] Example 4

[0079] Application of purine nucleoside compounds containing cinnamic acid in anti - plant bacteria. Here, the activities against Xanthomonas oryzae pv. oryzae (Xoo), Xanthomonas axonopodis pv. citri (Psa), and Pseudomonas syringae pv. actinidiae (Xac) are taken as examples.

[0080] The turbidimetric method was used to test the antibacterial activities of purine nucleoside compounds of cinnamic acid against Xanthomonas citri subsp. citri, Pseudomonas syringae pv. actinidiae, and Xanthomonas oryzae pv. oryzae. NA solid medium was used: 10 g of glucose, 5 g of peptone, 1 g of yeast, 3 g of beef extract, 15 g of agar, and 1000 mL of secondary water. Xanthomonas citri subsp. citri, Pseudomonas syringae pv. actinidiae, and Xanthomonas oryzae pv. oryzae were activated and placed in an incubator at 28 °C until single colonies grew. An appropriate amount of yellow single colonies was selected with an inoculation loop and placed into a conical flask containing a medium of NB: 10 g of glucose, 5 g of peptone, 1 g of yeast, 3 g of beef extract, and 1000 mL of secondary water, and shaken and cultured in a constant-temperature shaker until the logarithmic growth phase for standby.

[0081] The drugs to be tested were formulated to the specified concentrations. 1 mL of each was taken and added to a test tube containing 4 mL of NB medium, and then 40 μL of NB medium containing Xanthomonas oryzae pv. oryzae, Xanthomonas citri subsp. citri, and Pseudomonas syringae pv. actinidiae was added to the test tube. It was shaken and cultured in a constant-temperature shaker at 26 - 28 °C and 180 r / min. When the OD 595 value of the blank control group of NB liquid medium was 0.6 - 0.8, the OD 595 values of the bacterial suspensions at each concentration were measured on a spectrophotometer.

[0082] Corrected OD 595 value = OD of the bacterial-containing medium 595 − OD of the sterile medium 595

[0083] Inhibition rate (%) = (OD of the bacterial suspension in the corrected control medium 595 − OD of the drug-containing medium after correction 595 ) / OD of the bacterial suspension in the corrected control medium × 100%.

[0084] Table 5 In vitro antibacterial activities of some target compounds against plant pathogens Xoo, Psa, and Xac at a concentration of 50 μg / mL

[0085]

[0086] From Table 5, some compounds showed good antibacterial activities against Xanthomonas oryzae pv. oryzae (Xoo). At a concentration of 50 μg / mL, the inhibition activities of compounds 7, 20, and 21 against Xoo were 36.2%, 40.3%, and 35.6% respectively (superior to the control agent thiodiazole copper at 32.6%). When the concentration was below 50 μg / mL, the inhibition effects of compounds 1, 3, 23, and 24 against Psa were 21.3%, 20.4%, 27.8%, and 22.6% respectively, all superior to the control agent thiodiazole copper (18.6%). From the above, it can be seen that purine nucleoside compounds containing cinnamic acid structure have good antibacterial activities.

[0087] The above are only the preferred specific embodiments of the present invention, but the protection scope of the present invention is not limited thereto. Any changes or substitutions that can be easily thought of by those skilled in the art within the technical scope disclosed by the present invention should be covered by the protection scope of the present invention. Therefore, the protection scope of the present invention should be subject to the protection scope of the claims.

Claims

1. A class of purine nucleoside compounds containing cinnamic acid structure, characterized in that: It has a structure shown in the general formula (I): Wherein: R is independently selected from hydrogen, deuterium, optionally substituted or unsubstituted alkyl, optionally substituted or unsubstituted alkenyl, optionally substituted or unsubstituted alkynyl, optionally substituted or unsubstituted alkoxy, optionally substituted or unsubstituted cycloalkyl, optionally substituted or unsubstituted aryl or optionally substituted or unsubstituted heteroaryl.

2. A class of purine nucleoside compounds containing cinnamic acid structure according to claim 1, characterized in that: R is selected from 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.

3. A class of purine nucleoside compounds containing a cinnamic acid structure according to claim 1 or 2, characterized in that: The term "substituted" means substituted by one or more of C1-C6 alkyl, C1-C6 alkoxy, amino, hydroxy, halogen, nitro and trifluoromethyl.

4. The class of purine nucleoside compounds containing cinnamic acid structure according to claim 3, wherein: R is independently 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, phenyl, 2-methoxyphenyl, 3-methoxyphenyl, 4-methoxyphenyl, 2,3-dimethoxyphenyl, 3,4-dimethoxyphenyl, 3,4,5-trimethoxyphenyl, 2-hydroxyphenyl, 4-hydroxyphenyl, 4-methylphenyl, 4-diethylaminophenyl, benzo(1,3)dioxolyl, 4-fluorophenyl, 4-trifluorophenyl, 2-chlorophenyl, 4-chlorophenyl, 2,4-dichlorophenyl, 3,4-dichlorophenyl, 3-bromophenyl, 4-bromophenyl, 4-cyanophenyl, 2-nitrophenyl, 3-nitrophenyl, 4-nitrophenyl, 3-pyridyl, 2-furyl, 4-methylthiazolyl, 2-thienyl.

5. A class of purine nucleoside compounds containing a cinnamic acid structure according to claim 4, characterized in that, Selected from the following compounds: R is independently selected from phenyl, 2-methoxyphenyl, 3-methoxyphenyl, 4-methoxyphenyl, 2,3-dimethoxyphenyl, 3,4-dimethoxyphenyl, 3,4,5-trimethoxyphenyl, 2-hydroxyphenyl, 4-hydroxyphenyl, 4-methylphenyl, 4-diethylaminophenyl, benzo(1,3)dioxolyl, 4-fluorophenyl, 4-trifluorophenyl, 2-chlorophenyl, 4-chlorophenyl, 2,4-dichlorophenyl, 3,4-dichlorophenyl, 3-bromophenyl, 4-bromophenyl, 4-cyanophenyl, 2-nitrophenyl, 3-nitrophenyl, 4-nitrophenyl, 3-pyridyl, 2-furyl, 4-methylthiazolyl, 2-thienyl.

6. A method for preparing a purine nucleoside compound containing a cinnamic acid structure according to any one of claims 1-5, characterized in that: Its synthesis equation is as follows:

7. A pharmaceutical composition, characterized in that, Comprising the cinnamic acid-containing purine nucleoside compound according to any one of claims 1-5, or its stereoisomer, or its salt, or its solvate, and an agriculturally acceptable adjuvant or antiviral agent, insecticide or herbicide.

8. Use of the purine nucleoside compound containing cinnamic acid structure according to any one of claims 1-5 or the pharmaceutical composition according to claim 7 in preventing and controlling agricultural pests and diseases, characterized in that: The agricultural pests and diseases are viral diseases, bacterial diseases, fungal diseases or mite diseases.

9. The application in preventing and controlling agricultural pests and diseases according to claim 8, characterized in that: Viral diseases are tobacco mosaic virus, cucumber mosaic virus, and potato virus Y; bacterial diseases are bacterial leaf blight of rice, citrus canker, tobacco bacterial wilt, kiwifruit canker, and bacterial leaf streak of rice; fungal diseases are cucumber gray mold, pepper fusarium wilt, sclerotinia sclerotiorum of rapeseed, wheat scab, potato late blight, blueberry root rot, botryosphaeria dothidea, pitaya anthracnose, and sheath blight of rice; insect diseases are green worms, caterpillars, aphids, scale insects, and whiteflies; nematode diseases are root-knot nematodes, dagger nematodes, and stem nematodes; mite diseases are cotton leaf mites and citrus red spiders.

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