A 1-substituted cinnamoyl-2-oxo-zephyranthine compound, a preparation method and application thereof

Abstract

The application provides a 1-substituted cinnamoyl-2-oxo lycorine compound, a preparation method and application thereof. The structure of the compound is shown in the following formula I: wherein the benzene ring has one or more R groups which are substituents on the benzene ring, and the R group is selected from hydrogen, C 1‑6 alkyl, C 1‑6 alkylalkoxy, halogen, halogenated C 1‑6 alkyl, halogenated C 1‑6 alkoxy, nitro and dioxo. The compound shows excellent antiviral effect, especially for herpes simplex virus type I (HSV-1), and the compound has better safety.

Description

Technical Field

[0001] This application relates to the field of biomedicine, specifically to a 1-substituted cinnamyl-2-oxolycorine compound, its preparation method, and its application. Background Technology

[0002] Any discussion of prior art throughout the specification should not be construed as an admission that such prior art is well-known or constitutes part of common general knowledge in the art.

[0003] Traditional Chinese medicine (TCM) is a treasure of Chinese civilization. The natural compounds derived from traditional Chinese medicine and medicinal plants possess structural diversity, low toxicity, and wide availability, giving them unique advantages and enormous potential in preventing and treating viral infections. Lycorine, a TCM herb, is the bulb of the Lycoris radiata plant (family Amaryllidaceae). As a traditional medicinal plant, it has a long history of clinical application. Lycorine, the most important active ingredient of Lycoris radiata, was first isolated from the daffodil (Narcissus pseudonarcissus) in 1877 and has since been found to be widely distributed among Amaryllidaceae plants. Research on it has continued for nearly 150 years. Lycorine belongs to the isoquinoline alkaloid class, and its structure is as follows.

[0004]

[0005] Herpes simplex virus (HSV) is a double-stranded DNA virus with humans as its sole host. Infection is widespread in the human population and easily leads to latent infection. Epidemiological surveys indicate that HSV infection rates are nearly 90% globally. HSV-1 infection can cause oral and corneal herpes, and the increasingly prevalent genital herpes; in severe cases, it can even lead to encephalitis. Furthermore, it can remain latent in the nervous system for a long time and cause recurrent infections under stress or when the immune system is weakened. Currently, clinically used anti-HSV-1 drugs include nucleoside analogues such as acyclovir and ganciclovir, which effectively inhibit HSV-1 viral replication. For serious diseases caused by HSV, such as viral keratitis and viral encephalitis, these drugs significantly improve patient survival rates. However, most patients experience permanent neurological sequelae after recovery, including cognitive, memory, and behavioral impairments, and an increased incidence of epilepsy. Due to the widespread use of these drugs, antiviral agents are highly susceptible to mutations in their therapeutic target, the TK gene, leading to drug resistance. Given the limitations imposed by drug resistance and toxic side effects, the development and research of new anti-HSV-1 drugs is particularly important. In recent years, Traditional Chinese Medicine (TCM) has made outstanding contributions to antiviral research, and its characteristics of targeting multiple sites, acting on multiple pathways, and being less prone to drug resistance have led to its increasing acceptance. Summary of the Invention

[0006] This invention provides a 1-substituted cinnamyl-2-oxolycorine compound, its preparation method, and its applications. Bioactivity tests show that these compounds generally exhibit excellent inhibitory activity against herpes simplex virus type I (HSV-1). Furthermore, their inhibitory activity is not only significantly superior to lycorine, but also superior to lycorine derivatives where the 2-hydroxyl group is oxidized while the 1-hydroxyl group is not esterified. More encouragingly, these compounds also exhibit better safety, demonstrating promising potential for anti-HSV-1 drug applications.

[0007] Specifically, the present invention provides the following technical solutions.

[0008] In a first aspect of the invention, a 1-substituted cinnamyl-2-oxolimoline compound or a pharmaceutically acceptable salt thereof or a solvate thereof is provided, the structure of which is shown in Formula I:

[0009]

[0010] The benzene ring has one or more R groups, which are substituents on the benzene ring, wherein the R groups are selected from hydrogen, C, and D. 1-6 Alkyl, C 1-6 Alkoxy, halogen, halogenated C 1-6 Alkyl, Halogenated C1-6 Alkyl, nitro, and dioxane groups.

[0011] In some embodiments of the present invention, the benzene ring has an R group, wherein the R group is R1, and R1 is selected from hydrogen, C… 1-3 Alkyl, C 1-3 Alkoxy, halogen, halogenated C 1-3 Alkyl, Halogenated C 1-3 Alkyl, nitro, and dioxane groups.

[0012] In some embodiments of the present invention, the benzene ring has two R groups, namely R3 and R4, wherein R3 and R4 are each independently selected from C 1-3 Alkyl, C 1-3 Alkylalkoxy, halogen, halogenated C 1-3 Alkyl, Halogenated C 1-3 Alkyl groups and nitro groups.

[0013] In these embodiments, the substitution positions of R3 and R4 on the benzene ring are selected from combinations of the following positions: C-2 and C-4, C-2 and C-5, C-2 and C-6, and C-3 and C-4.

[0014] In embodiments of the present invention, alkyl refers to a saturated aliphatic group consisting only of carbon and hydrogen, with the general formula C0. n H 2n+1 C 1-6 Alkyl groups refer to alkyl groups with 1-6 carbon atoms, such as the C16 group described in this invention. 1-6 Alkyl groups include methyl (-CH3), ethyl (-C2H5), propyl (-C3H7), butyl (-C4H9), and pentyl (-C5H5). 11 ), hexyl (-C6H) 13 ).

[0015] C 1-6 An alkoxy group represents a substituent consisting of an alkyl group containing 1-6 carbon atoms and an oxygen atom, such as the C group described in this invention. 1-6 Alkoxy groups can include methoxy (CH3O-), ethoxy (C2H5O-), propoxy (C3H7O-), n-propoxy (CH3CH2CH2O-), isopropoxy (CH3CH(CH3)O-), butoxy (C4H9O-), n-butoxy (CH3CH2CH2CH2O-), isobutoxy (CH3CH(CH3)CH2O-), sec-butoxy (CH3CH2CH(CH3)O-), tert-butoxy (CH(CH3)CH(CH3)O-), and pentoxy (C5H 11O-), n-pentoxy (CH3CH2CH2CH2CH2O-), isopentoxy (CH(CH)CH(CH)CH(O-)), secondary pentoxy (CH(CH)CH(CH)O-), tertiary pentoxy (CH(CH)CH(O-)), neopentoxy (CH(CH)O-), hexyloxy (C6H) 13 O-), n-hexyloxy (CH3CH2CH2CH2CH2CH2O-), isohexyloxy (CH(CH)CH(CH)CH(CH)O-), sec-hexyloxy (CH(CH)CH(CH)O-), tertiary hexyloxy (CH(CH)O-), etc.

[0016] Halogens are selected from F, Cl, Br, I, etc., and are particularly selected from F, Cl, and Br in this invention.

[0017] Halogenated C 1-6 An alkyl group is a substituent consisting of an alkyl group containing 1-6 carbon atoms and a halogen atom (fluorine, chlorine, bromine, iodine, etc.), for example, the halogenated C group described in this invention. 1-6 Alkyl groups can include fluoromethyl (-CH2F), trifluoromethyl (-CF3), chloroethyl (-C2H4Cl), bromopropyl (-C3H6Br), and so on.

[0018] Halogenated C 1-6 An alkoxy group is a substituent consisting of an alkoxy group containing 1-6 carbon atoms and a halogen atom (fluorine, chlorine, bromine, iodine), such as the halogenated C group described in this invention. 1-6 Alkoxy groups can include fluoromethoxy (-OCH2F), chloroethoxy (-OC2H4Cl), trifluoromethoxy (-OCF3), and so on.

[0019] In some embodiments of the present invention, R1 is preferably F, Br, methoxy, trifluoromethyl, trifluoromethoxy, nitro, and dioxane.

[0020] In some embodiments of the invention, R3 and R4 are each independently selected from F, Cl, Br, and nitro. In these embodiments, R3 and R4 may be the same or different.

[0021] In some embodiments of the present invention, R1 is selected from fluorine, bromine, trifluoromethoxy, and nitro;

[0022] R3 and R4 are each independently selected from fluorine, chlorine, bromine, and nitro; R3 and R4 may be the same or different;

[0023] The substitution positions of R3 and R4 on the benzene ring are selected from combinations of the following positions: C-2 and C-5, C-2 and C-6, C-3 and C-4.

[0024] Furthermore, the present invention provides some compounds as examples; specifically, the compounds described in the present invention may include:

[0025] S1: 1-(2-Fluorocinyl)-2-oxolimoline;

[0026] S2: 1-(4-Fluorocinyl)-2-oxolimoline;

[0027] S3: 1-(3,4-difluorocinnamyl)-2-oxolycorine;

[0028] S4: 1-(2-trifluoromethylcinnamoyl)-2-oxolimoline;

[0029] S5: 1-(4-trifluoromethylcinnamoyl)-2-oxolimoline;

[0030] S6: 1-(3-trifluoromethoxycinnamoyl)-2-oxolimoline;

[0031] S7: 1-(2,5-dichlorocinnamyl)-2-oxolycorine;

[0032] S8: 1-(2,6-dichlorocinnamyl)-2-oxolycorine;

[0033] S9: 1-(4-bromocinnamyl)-2-oxolycorine;

[0034] S10: 1-(3-nitrocinnamyl)-2-oxolycorine;

[0035] S11: 1-(2-chloro-5-nitrocinnamyl)-2-oxolimoline;

[0036] S12: 1-(2-nitro-5-bromocinnamoyl)-2-oxolimoline;

[0037] S13: 1-(2,4-dinitrocinnamyl)-2-oxolimoline;

[0038] S14: 1-(4-methoxycinnamoyl)-2-oxolimoline;

[0039] S15: 1-(3,4-dioxonylcinnamyl)-2-oxolimoline; and

[0040] S16: 1-Cinnamyl-2-oxolimoline.

[0041] The pharmaceutically acceptable salts described in this invention refer to acidic salts formed by the 1-substituted cinnamyl-2-oxolycorine compounds of this invention with inorganic and / or organic acids. They may also include zwitterionic salts (internal salts) and quaternary ammonium salts, such as alkylammonium salts. These salts can be obtained directly during the final separation and purification of the compounds. Alternatively, they can be obtained by mixing the aforementioned compounds with an appropriate amount (e.g., equimolar amounts) of acid. These salts may precipitate in solution and be collected by filtration, or be recovered after solvent evaporation, or be prepared by freeze-drying after reaction in an aqueous medium. In some embodiments of this invention, the pharmaceutically acceptable salts described in this invention may be hydrochlorides, hydrobromates, sulfates, hydrogen sulfates, nitrates, phosphates, hydrogen phosphates, formates, acetates, propionates, benzoates, succinates, fumarates, maleates, lactates, citrates, tartrates, succinates, gluconates, methanesulfonates, ethylsulfonates, benzenesulfonates, p-toluenesulfonates, etc.

[0042] A solvate is a compound of the present invention that forms a complex by coordination with a solvent molecule, and it may be in a solid or liquid state. A hydrate is a specific form of solvate that coordinates with water. The scope of solvates described in this invention includes hydrates. In some embodiments of the invention, the solvate includes a solvate of the compound of Formula I with any one of water, ethanol, isopropanol, and acetone.

[0043] In a second aspect of the invention, a method is provided for preparing the 1-substituted cinnamyl-2-oxolycorine compounds as described above, or pharmaceutically acceptable salts thereof, or solvates thereof, comprising:

[0044] Starting with lycorine, the hydroxyl groups at the 1 and 2 positions of lycorine were acetylated with acetic anhydride to obtain the intermediate INB.

[0045] Intermediate INB is selectively deacetylated at position 2 under acidic conditions to yield intermediate INC;

[0046] The 2-hydroxyl group of intermediate INC is oxidized to obtain intermediate IND;

[0047] The intermediate IND is deacetylated at position 1 under acidic conditions to give the intermediate INE.

[0048] intermediate INE and An esterification reaction occurs in the presence of a condensing agent to obtain 1-substituted cinnamyl-2-oxolimoline compounds as shown in Formula I.

[0049] Where R is the same as defined in the first aspect above;

[0050]

[0051] For example, in some specific embodiments, the preparation method includes:

[0052] In the presence of the acid-binding agent pyridine, the hydroxyl groups at the 1 and 2 positions of lycorine are selectively protected with acetic anhydride to yield the intermediate INB.

[0053] Intermediate INB was selectively deacetylated at position 2 under heating conditions in the presence of concentrated hydrochloric acid to obtain intermediate INC;

[0054] The 2-hydroxyl group of intermediate INC is oxidized at low temperature in the presence of an oxidizing agent to obtain intermediate IND;

[0055] The intermediate IND was deacetylated at position 1 under heating conditions in the presence of concentrated hydrochloric acid to obtain the intermediate INE.

[0056] The intermediate INE undergoes esterification with R-substituted cinnamic acid in the presence of a condensing agent and a catalyst.

[0057] In a third aspect of the invention, a pharmaceutical composition is provided comprising the 1-substituted cinnamyl-2-oxolycorine compound described in the first aspect above, or a pharmaceutically acceptable salt thereof, or a solvate thereof.

[0058] In a fourth aspect of the invention, a pharmaceutical formulation is provided comprising the 1-substituted cinnamyl-2-oxolycorine compound described in the first aspect above, or a pharmaceutically acceptable salt thereof or a solvate thereof, and at least one pharmaceutically acceptable excipient or pharmaceutical carrier.

[0059] The pharmaceutical compositions or pharmaceutical preparations involved in this invention may contain one or more of the 1-substituted cinnamyl-2-oxolimoline compounds described in this invention.

[0060] In some embodiments of the present invention, the pharmaceutical composition or pharmaceutical preparation of the present invention can be administered via the gastrointestinal tract or non-gastrointestinal tract, and the dosage form includes, but is not limited to, tablets, capsules, pills and injections.

[0061] Commonly used pharmaceutically acceptable excipients include binders, fillers, wetting agents, and disintegrants, which can be used in oral tablets, capsules, and pills. If necessary, other excipients such as flavoring agents, coloring agents, stabilizers, and lubricants can also be added, or suitable dosage forms can be coated using pharmaceutically known methods.

[0062] Some embodiments of the present invention provide methods for producing pharmaceutical compositions or pharmaceutical formulations, the methods comprising mixing one or more of the 1-substituted cinnamyl-2-oxolimanesine compounds of the present invention or their pharmaceutically acceptable salts or solvates with pharmaceutical excipients or pharmaceutical carriers. For example, in some embodiments, solid or liquid formulations can be prepared by uniformly mixing the active compound with liquid and / or finely pulverized solid excipients in desired proportions, and then, if necessary, shaping the resulting mixture into the desired form. Parenteral dosage forms are prepared, for example, by dissolving the compound of the present invention in a suitable liquid medium, filtering and sterilizing the solution, and then filling and sealing it into suitable vials or ampoules.

[0063] Of course, in addition to these methods, those skilled in the art can also use other techniques known in the art to formulate the compounds of the present invention into pharmaceutical compositions or pharmaceutical preparations. For example, pharmaceutical preparations can be prepared according to the Modern Pharmaceutical Preparations Series edited by Shenyang Pharmaceutical University. Furthermore, besides those mentioned in this invention, suitable pharmaceutical excipients can also be other types known in the art, such as those described in the Handbook of Parmaceutical Excipients, edited by Paul J. Sheskey et al., which is currently in its eighth edition; the first edition was published in 1986, and the eighth edition was first published in 2017.

[0064] In a fifth aspect of the invention, the use of the 1-substituted cinnamyl-2-oxolycorine compounds described in the first aspect above, or pharmaceutically acceptable salts thereof or solvates thereof, or pharmaceutical compositions or pharmaceutical preparations comprising such compounds, or 2-oxolycorine in the preparation of antiviral drugs is provided.

[0065] In embodiments of the present invention, the virus is, in particular, herpes simplex virus type I.

[0066] In a sixth aspect of the invention, the use of the 1-substituted cinnamyl-2-oxolycorine compounds described in the first aspect above, or pharmaceutically acceptable salts thereof or solvates thereof, or pharmaceutical compositions or pharmaceutical preparations comprising such compounds, or 2-oxolycorine in the preparation of medicaments for treating diseases associated with herpes simplex virus type I infection is provided.

[0067] In embodiments of the present invention, diseases associated with type I herpes simplex virus include oral herpes or cold herpes, eye infections (such as keratitis, conjunctivitis and retinitis caused by HSV-1), genital infections, and nervous system infections (such as meningitis and encephalitis caused by HSV-1).

[0068] In a seventh aspect of the invention, a method for treating herpes simplex virus type I infection is provided, comprising administering to a subject a 1-substituted cinnamyl-2-oxolycorine compound as described in the first aspect of the invention above, or a pharmaceutically acceptable salt thereof or a solvate thereof, or a pharmaceutical composition or pharmaceutical preparation containing such a compound.

[0069] In an eighth aspect of the invention, a method for treating a disease associated with herpes simplex virus type I infection is provided, comprising administering to a subject a 1-substituted cinnamyl-2-oxolycorine compound as described in the first aspect of the invention above, or a pharmaceutically acceptable salt thereof or a solvate thereof, or a pharmaceutical composition or pharmaceutical preparation comprising such a compound.

[0070] The subject in this invention refers to an animal that is already being treated, observed, or experimented on, preferably a mammal, and most preferably a human.

[0071] The treatment described in this invention includes alleviating or partially alleviating the symptoms of the treated disease, syndrome, symptom, or disorder.

[0072] The optimal dosage and interval of the compounds described in this invention are determined by the properties of the compounds and external conditions such as the form, route and site of administration, and the specific mammal being treated, and this optimal dosage can be determined using conventional techniques. It should also be recognized that the optimal course of treatment, i.e., the daily dose of the compound over a specified period, can be determined using methods known in the art.

[0073] Although the dosage varies with symptoms and the patient's age, the nature and severity of the disease or disorder, and the route and manner of administration, for reference when administering the compound orally to adult patients, the normal dosage of the compound is 1 to 1000 mg, preferably 5 to 500 mg, per day, in the form of a single dose or divided doses, for example, twice or three times daily; for intravenous administration, the dosage may be divided into one to three doses of 0.1 to 100 mg, preferably 0.5 to 50 mg, per day.

[0074] Compared to existing technologies, the advantages of this invention include:

[0075] This invention provides a class of 1-substituted cinnamyl-2-oxolycorine compounds represented by Formula I. Preliminary activity screening experiments revealed that these compounds exhibit good inhibitory activity against type I herpes simplex virus. Their antiviral activity is not only significantly better than that of lycorine, but also better than lycorine derivatives in which the 2-hydroxyl group is oxidized while the 1-hydroxyl group is not esterified. More encouragingly, these compounds show better safety and demonstrate promising potential for anti-type I herpes simplex virus treatment. Detailed Implementation

[0076] The present application is further illustrated below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the application. Experimental methods in the following embodiments that do not specify specific conditions are generally performed under conventional conditions or as recommended by the manufacturer.

[0077] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of skill in the art. All reagents and materials used in this application are readily available through conventional means, and unless otherwise specified, they are used in accordance with conventional methods in the art or as per the product instructions. Furthermore, any methods and materials similar to or equivalent to those described herein may be applied to the methods of this application. The preferred embodiments and materials described herein are for illustrative purposes only.

[0078] Unless otherwise specified, raw materials are generally available from commercial sources. Commercially available solvents or reagents are generally used without further purification. The structure of the compound is determined by nuclear magnetic resonance (NMR) spectroscopy. The shifts (δ) of the 1H and 1C NMR spectra are given in parts per million (ppm). Deuterated chloroform (CDCl3) or deuterated dimethyl sulfoxide (DMSO-d6) is used as the solvent for both 1H and 1C NMR spectra, with tetramethylsilane (TMS) as the internal standard. Column chromatography generally uses 200–300 mesh silica gel as the support and n-hexane-ethyl acetate as the elution solvent.

[0079] Illustrations of abbreviations used in this invention:

[0080] Ac2O: Acetic anhydride; Py: Pyridine; DMSO: Dimethyl sulfoxide; ClCOCOCl: Oxaloyl chloride; K2CO3: Anhydrous potassium carbonate; EDCI: 1-Ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride; DMAP: 4-Dimethylaminopyridine; HCl: Concentrated hydrochloric acid; THF: Tetrahydrofuran; DCM: Dichloromethane; CH3OH: Methanol.

[0081] This invention provides a class of 1-substituted cinnamyl-2-oxolimanesine compounds as shown in Formula I:

[0082]

[0083] The benzene ring has one or more R groups, which are substituents on the benzene ring, wherein the R groups are selected from hydrogen, C, and D. 1-6 Alkyl, C 1-6 Alkoxy, halogen, halogenated C 1-6 Alkyl, Halogenated C 1-6 Alkyl, nitro, and dioxane groups.

[0084] Methods for preparing these compounds are provided, including:

[0085] Starting with lycorine (INA), the hydroxyl groups at the 1 and 2 positions of lycorine were acetylated with acetic anhydride to obtain the intermediate INB.

[0086] Intermediate INB is selectively deacetylated at position 2 under acidic conditions to yield intermediate INC;

[0087] The 2-hydroxyl group of intermediate INC is oxidized to obtain intermediate IND;

[0088] The intermediate IND is deacetylated at position 1 under acidic conditions to give the intermediate INE.

[0089] intermediate INE and An esterification reaction occurs in the presence of a condensing agent to obtain 1-substituted cinnamyl-2-oxolimoline compounds as shown in Formula I.

[0090] Wherein, R is defined as in equation I;

[0091]

[0092] Specifically, the compound can be prepared according to the reaction route shown below:

[0093]

[0094] As an example, the present invention provides a series of compounds, and the specific preparation process and pharmaceutical activity of these compounds are shown in the following embodiments.

[0095] Examples of the preparation of compounds of the present invention

[0096] Example 1 1,2-Dacetyllycorine (INB)

[0097] INB structure:

[0098] Lycorine (15.85 g, 55.0 mmol) was suspended in pyridine (40.0 mL) in a 500 mL round-bottom flask, and acetic anhydride (47.5 mL, 475.0 mmol) was added with stirring at room temperature. The mixture was heated to 60 °C and stirred for 12 h. Then, MeOH (75.0 mL) was added to the reaction system, and the mixture was stirred for 1 h. The solvent was removed by concentration under reduced pressure. CH2Cl2 (250 mL) and saturated NaHCO3 solution (200 mL) were added to the residue sequentially. After stirring at room temperature for 10 min, the mixture was separated. The organic phase was washed with saturated NaCl solution (200 mL) and dried over anhydrous Na2SO4. The mixture was concentrated under reduced pressure, and silica gel chromatography yielded a white solid, INB (18.43 g, 91%). 1H NMR(500MHz, CDCl3)δ6.75(s,1H),6.57(s,1H),5.91(s,2H),5.73(s,1H),5.5 9–5.45(m,1H),5.37–5.19(m,1H),4.16(d,J=14.1Hz,1H),3.53(d,J=14.0Hz, 1H),3.43–3.29(m,1H),2.88(d,J=10.4Hz,1H),2.77(d,J=10.4Hz,1H),2.65( ddd,J=8.2,3.6,1.7Hz,2H),2.40(q,J=8.8Hz,1H),2.08(s,3H),1.95(s,3H); 13 C NMR (125MHz, CDCl3) δ169.98,169.73,146.42,146.30,146.14,129.43,126.55,113.79, 107.30,105.04,100.96,70.89,69.24,61.22,56.90,53.61,40.52,28.66,21.13,20.92.

[0099] Example 2 1-Acetyllycorine (INC)

[0100] INC structure:

[0101] In a 500 mL round-bottom flask, INB (8.45 g, 22.8 mmol) was suspended in methanol (200.0 mL), and concentrated hydrochloric acid (20.0 mL) was added with stirring at room temperature. The mixture was heated to 55 °C and stirred for 3 h. After cooling to room temperature, the solvent was removed by concentration under reduced pressure. Then, ethyl acetate (150 mL) and saturated NaHCO3 solution (200 mL) were added to the residue sequentially. After stirring at room temperature for 10 min, the mixture was separated. The organic phase was washed with saturated NaCl solution (200 mL) and dried over anhydrous Na2SO4. The solution was concentrated under reduced pressure, and silica gel chromatography was used to obtain a white solid INC (6.35 g, 84.8%). 1H NMR (500MHz, CDCl3) δ6.61(s,1H),6.56(s,1H),5.91(s,2H),5.58(s,1H),5.52(s,1H),4.20–4.03(m,2H),3.80(s,1H),3.49(d,J=13.9Hz ,1H),3.34(dt,J=9.2,4.7Hz,1H),2.84(d,J=10.5Hz,1H),2.74(d,J=10.5Hz,1H),2.68–2.54(m,2H),2.36(q,J=8.8Hz,1H),1.92(s,3H); 13 C NMR (125MHz, CDCl3) δ170.76,146.46,146.20,143.63,129.24,127.02,117.42,1 07.27,104.88,100.93,72.70,69.38,61.60,56.82,53.70,39.19,28.53,21.06.

[0102] Example 3 1-Acetyl-2-oxolycorine (IND)

[0103] IND structure:

[0104] In a 500 mL round-bottom flask, Tf₂O (6.9 mL, 40.8 mmol) was dissolved in anhydrous dichloromethane (50.0 mL). Under nitrogen protection, DMSO (4.8 mL, 61.2 mmol) was added dropwise with vigorous stirring at -45 °C. After the addition was complete, the reaction mixture was kept at this temperature and stirred for 0.5 h. Then, INC (6.71 g, 20.4 mmol) in anhydrous dichloromethane (100.0 mL) was slowly added dropwise. After the addition was complete, the reaction mixture was kept at this temperature and stirred for 2.5 h. Then, triethylamine (16.8 mL, 122.4 mmol) was added dropwise. The mixture was kept at this temperature and stirred for 10 min, and then allowed to warm to room temperature. Purified water (200.0 mL) was added dropwise to the reaction mixture. The mixture was separated, and the organic phase was washed with saturated NaCl solution (200 mL) and then dried over anhydrous Na₂SO₄. The solution was concentrated under reduced pressure and purified by silica gel chromatography to give a white solid IND (5.44 g, 81.5%). 1H NMR (500MHz, CDCl3) δ6.73(s,1H),6.58(s,1H),5.99(d,J=3.2Hz,2H),5.95–5.88(m,2H),4.17(d,J=14.1Hz,1H),3.61(d,J=14.0Hz,1H),3 .46(dt,J=9.0,4.4Hz,1H),3.26(d,J=9.9Hz,1H),3.17(d,J=10.0Hz,1H),2.86(dd,J=8.3,2.0Hz,2H),2.53(q,J=8.7Hz,1H),1.96(s,3H); 13 C NMR (125MHz, CDCl3) δ192.99,169.55,169.10,146.71,146.67,128.88,125.24,1 20.43,107.34,105.42,101.14,69.00,62.32,56.33,53.25,45.52,30.00,20.81.

[0105] Example 4 2-Oxylycorine (INE, S17)

[0106] INE structure:

[0107] IND (4.71 g, 14.4 mmol) was suspended in methanol (100.0 mL) in a 500 mL round-bottom flask, and concentrated hydrochloric acid (10.0 mL) was added with stirring at room temperature. The mixture was heated to 50 °C and stirred for 3 h. After cooling to room temperature, the solvent was removed by concentration under reduced pressure. Silica gel chromatography yielded a grayish-white solid INE (3.58 g, 87.3%). 1 H NMR (500MHz, DMSO) δ6.84(s,1H),6.71(s,1H),5.96(d,J=2.2Hz,2H),5.81(s,1H),4.41(d,J=2.6Hz,1H),4.06(d,J=14.3Hz,1H),3.44(d,J=14.2H z,1H),3.30(t,J=8.1Hz,1H),3.09(d,J=9.8Hz,1H),2.89(d,J=9.8Hz,1H ),2.81(dd,J=19.3,7.4Hz,1H),2.76–2.64(m,1H),2.38(q,J=8.6Hz,1H); 13C NMR (125MHz, DMSO) δ197.55,168.60,145.69,145.60,129.22,127.36,119.07,106.97,105.56,100.67,69.15,61.57,55.90,52.78,46.04,29.34.

[0108] Example 5 1-(2-Fluorocinnamyl)-2-oxolycorine (S1)

[0109] S1 structure:

[0110] In a 250 ml round-bottom reaction flask, INE (5.0 mmol) and 2-fluorocinnamic acid (6.0 mmol) were dissolved in 50 ml of anhydrous dichloromethane. Under nitrogen protection at room temperature, EDCI·HCl (9.0 mmol) and DMAP (0.5 mmol) were added sequentially. After the addition was complete, the reaction mixture was stirred and kept at 35 °C. HPLC monitoring showed that the remaining INE content was less than 5%, at which point further processing was possible. The reaction solution was washed once with 100 ml of purified water and once with 100 ml of 10% sodium chloride solution, and concentrated under reduced pressure by column chromatography to obtain a white solid, S1.

[0111] Example 6 1-(4-Fluorocinyl)-2-oxolycorine (S2)

[0112] S2 structure:

[0113] In a 250 ml round-bottom reaction flask, INE (5.0 mmol) and 4-fluorocinnamic acid (6.0 mmol) were dissolved in 50 ml of anhydrous dichloromethane. Under nitrogen protection at room temperature, EDCI·HCl (9.0 mmol) and DMAP (0.5 mmol) were added sequentially. After the addition was complete, the reaction mixture was stirred and kept at 35 °C. HPLC monitoring showed that the remaining INE content was less than 5%, at which point further processing was possible. The reaction solution was washed once with 100 ml of purified water and once with 100 ml of 10% sodium chloride solution, and concentrated under reduced pressure by column chromatography to obtain a white solid, S2. 1H NMR (400MHz, CDCl3) δ7.60 (d, J=16.0Hz, 1H), 7.44 (dd, J=8.6, 5.4Hz, 2H), 7.03 (t, J=8.6Hz, 2 H),6.80(s,1H),6.57(s,1H),6.20(d,J=16.0Hz,1H),6.14(d,J=3.1Hz,1H),6.03(d,J=1.8Hz ,1H),5.90(d,J=4.2Hz,2H),4.19(d,J=14.1Hz,1H),3.63(d,J=14.1Hz,1H),3.49(dt,J=8.9, 4.5Hz,1H),3.33(d,J=10.3Hz,1H),3.26(d,J=9.6Hz,1H),2.89(s,2H),2.56(q,J=8.7Hz,1H); 13 C NMR (100MHz, CDCl3) δ192.92,168.98,165.46,146.74,146.70,144.75,130.11,130.03,128.80,125.21,12 0.52,116.83,116.81,116.13,115.91,107.31,105.52,101.10,69.06,62.42,56.33,53.27,45.62,30.02.

[0114] Example 7 1-(3,4-Difluorocinnamyl)-2-oxolycorine (S3)

[0115] S3 structure:

[0116] In a 250 mL round-bottom reaction flask, INE (5.0 mmol) and 3,4-difluorocinnamic acid (6.0 mmol) were dissolved in 50 mL of anhydrous dichloromethane. Under nitrogen protection at room temperature, EDCI·HCl (9.0 mmol) and DMAP (0.5 mmol) were added sequentially. After the addition was complete, the reaction mixture was stirred and kept at 35 °C. HPLC monitoring showed that the remaining INE content was less than 5%, at which point further processing was possible. The reaction solution was washed once with 100 mL of purified water and once with 100 mL of 10% sodium chloride solution, and concentrated under reduced pressure by column chromatography to obtain a white solid, S3. 1H NMR (600MHz, CDCl3) δ7.54(d,J=15.9Hz,1H),7.27(dd,J=3.8,1.5Hz,1H),7.25(d,J=1.7Hz,0H),7.18( s,1H),7.13(dd,J=17.6,8.4Hz,1H),6.78(s,1H),6.57(s,1H),6.19(d,J=15.9Hz,1H),6.13(d,J=3.2H z,1H),6.03(d,J=1.6Hz,1H),5.90(s,1H),5.90(s,1H),4.19(d,J=14.1Hz,1H),3.63(d,J=13.7Hz,1H) ,3.50–3.46(m,1H),3.33(d,J=10.0Hz,1H),3.24(d,J=10.0Hz,1H),2.89(m,2H),2.56(q,J=8.8Hz,1H); 13 C NMR (150MHz, CDCl3) δ192.79,169.09,165.10,146.75,146.70,143.64,128.84,125.13,120.48,118.23,11 8.22,117.86,117.75,116.39,116.27,107.34,105.45,101.11,69.23,62.40,56.32,53.26,45.60,30.02.

[0117] Example 8 1-(2-Trifluoromethylcinnamoyl)-2-oxolycorine (S4)

[0118] S4 structure:

[0119] In a 250 mL round-bottom reaction flask, INE (5.0 mmol) and 2-trifluoromethylcinnamic acid (6.0 mmol) were dissolved in 50 mL of anhydrous dichloromethane. Under nitrogen protection at room temperature, EDCI·HCl (9.0 mmol) and DMAP (0.5 mmol) were added sequentially. After the addition was complete, the reaction mixture was stirred and kept at 35 °C. HPLC monitoring showed that the remaining INE content was less than 5%, at which point further processing was possible. The reaction solution was washed once with 100 mL of purified water and once with 100 mL of 10% sodium chloride solution, and concentrated under reduced pressure by column chromatography to obtain a white solid, S4. 1H NMR (600MHz, CDCl3) δ7.96(dd,J=15.8,1.8Hz,1H),7.66(d,J=7.7Hz,1H),7.62(d,J=7.8Hz,1H),7.51(t,J=7.6Hz ,1H),7.45(t,J=7.5Hz,1H),6.78(s,1H),6.57(s,1H),6.27(d,J=15.8Hz,1H),6.11(d,J=3.1Hz,1H),6.03(d,J=1 .8Hz,1H),5.91(d,J=1.3Hz,1H),5.89(d,J=1.3Hz,1H),4.19(d,J=14.0Hz,1H),3.61(d,J=14.0Hz,1H),3.48(ddd ,J=9.0,5.3,3.6Hz,1H),3.34(d,J=10.0Hz,1H),3.25(d,J=9.8Hz,1H),2.92–2.81(m,2H),2.54(q,J=8.6Hz,1H); 13 C NMR (150MHz, CDCl3) δ192.72,169.07,164.57,146.77,146.73,141.20,132.97,131.99,129.77,128.86,12 7.79,126.20,126.16,125.05,121.26,120.46,107.31,105.39,101.12,69.49,62.54,56.41,53.33,45.72 30.01.

[0120] Example 9 1-(4-trifluoromethylcinnamoyl)-2-oxolycorine (S5)

[0121] S5 structure:

[0122] In a 250 mL round-bottom reaction flask, INE (5.0 mmol) and 4-trifluoromethylcinnamic acid (6.0 mmol) were dissolved in 50 mL of anhydrous dichloromethane. Under nitrogen protection at room temperature, EDCI·HCl (9.0 mmol) and DMAP (0.5 mmol) were added sequentially. After the addition was complete, the reaction mixture was stirred and kept at 35 °C. HPLC monitoring showed that the remaining INE content was less than 5%, at which point further processing was possible. The reaction solution was washed once with 100 mL of purified water and once with 100 mL of 10% sodium chloride solution, and concentrated under reduced pressure by column chromatography to obtain a white solid, S5. 1H NMR (400MHz, CDCl3) δ7.65(d,J=16.0Hz,1H),7.60(d,J=8.2Hz,2H),7.55(d,J=8.2Hz, 2H),6.79(s,1H),6.57(s,1H),6.35(d,J=16.0Hz,1H),6.15(d,J=2.6Hz,1H),6.04(s,1 H),5.90(d,J=5.2Hz,2H),4.19(d,J=14.2Hz,1H),3.63(d,J=14.1Hz,1H),3.53–3.44(m ,1H),3.35(d,J=9.9Hz,1H),3.26(d,J=9.8Hz,1H),2.90(s,2H),2.57(q,J=8.6Hz,1H); 13 C NMR (100MHz, CDCl3) δ192.74,169.11,165.05,146.78,144.13,137.51,128.28,125.8 2,120.50,119.67,107.35,105.46,101.12,69.32,62.40,56.33,53.27,45.62,30.04.

[0123] Example 10 1-(3-Trifluoromethoxycinnamoyl)-2-oxolycorine (S6)

[0124] S6 structure:

[0125] In a 250 mL round-bottom reaction flask, INE (5.0 mmol) and 3-trifluoromethoxycinnamic acid (6.0 mmol) were dissolved in 50 mL of anhydrous dichloromethane. Under nitrogen protection at room temperature, EDCI·HCl (9.0 mmol) and DMAP (0.5 mmol) were added sequentially. After the addition was complete, the reaction mixture was stirred and kept at 35 °C. HPLC monitoring showed that the remaining INE was less than 5%, at which point further processing was possible. The reaction solution was washed once with 100 mL of purified water and once with 100 mL of 10% sodium chloride solution, and concentrated under reduced pressure by column chromatography to obtain a white solid, S6.

[0126] Example 11 1-(2,5-Dichlorocinnamyl)-2-oxolycorine (S7)

[0127] S7 structure:

[0128] In a 250 mL round-bottom reaction flask, INE (5.0 mmol) and 2,5-dichlorocinnamic acid (6.0 mmol) were dissolved in 50 mL of anhydrous dichloromethane. Under nitrogen protection at room temperature, EDCI·HCl (9.0 mmol) and DMAP (0.5 mmol) were added sequentially. After the addition was complete, the reaction mixture was stirred and kept at 35 °C. HPLC monitoring showed that the remaining INE was less than 5%, at which point further processing was possible. The reaction solution was washed once with 100 mL of purified water and once with 100 mL of 10% sodium chloride solution, and concentrated under reduced pressure by column chromatography to obtain a white solid, S7. 1 H NMR (600MHz, CDCl3) δ7.95(d,J=16.0Hz,1H),7.50(d,J=2.4Hz,1H),7.31(d,J=8.6Hz,1H),7.24(dd,J= 8.6,2.4Hz,1H),6.78(s,1H),6.57(s,1H),6.27(d,J=15.9Hz,1H),6.12(d,J=3.1Hz,1H),6.03(d,J=1.9 Hz,1H),5.91(d,J=1.3Hz,1H),5.91(d,J=1.3Hz,1H),4.19(d,J=14.1Hz,1H),3.62(d,J=14.0Hz,1H),3. 51–3.46(m,1H),3.34(d,J=9.9Hz,1H),3.25(d,J=9.8Hz,1H),2.93–2.84(m,2H),2.55(q,J=8.7Hz,1H); 13 C NMR (150MHz, CDCl3) δ192.67,169.10,164.72,146.78,146.73,140.49,133.86,133.20,132.99,131.26,131.0 0,128.85,127.39,125.06,120.82,120.46,107.35,105.39,101.12,69.45,62.44,56.36,53.29,45.63,30.01.

[0129] Example 12 1-(2,6-Dichlorocinnamyl)-2-oxolycorine (S8)

[0130] S8 structure:

[0131] In a 250 mL round-bottom reaction flask, INE (5.0 mmol) and 2,6-dichlorocinnamic acid (6.0 mmol) were dissolved in 50 mL of anhydrous dichloromethane. Under nitrogen protection at room temperature, EDCI·HCl (9.0 mmol) and DMAP (0.5 mmol) were added sequentially. After the addition was complete, the reaction mixture was stirred and kept at 35 °C. HPLC monitoring showed that the remaining INE content was less than 5%, at which point further processing was possible. The reaction solution was washed once with 100 mL of purified water and once with 100 mL of 10% sodium chloride solution, followed by concentrated solution under reduced pressure and column chromatography to obtain a white solid, S8. 1 H NMR(400MHz, CDCl3)δ7.75(d,J=16.4Hz,1H),7.32(s,1H),7.30(s,1H),7.21–7.11(m,1H) ,6.79(s,1H),6.58(s,1H),6.50(d,J=16.4Hz,1H),6.13(d,J=2.6Hz,1H),6.04(s,1H),5.9 2(d,J=1.2Hz,1H),5.91(d,J=1.2Hz,1H),4.19(d,J=14.2Hz,1H),3.64(d,J=13.1Hz,1H), 3.54–3.44(m,1H),3.36(d,J=9.3Hz,1H),3.29(s,1H),2.90(s,2H),2.57(d,J=6.8Hz,1H); 13 CNMR (100MHz, CDCl3) δ164.96,146.79,139.37,135.12,131.42,130.00,128.83,125.3 6,125.10,120.59,107.32,105.43,101.14,69.48,62.40,56.26,53.29,45.59,30.01.

[0132] Example 13 1-(4-bromocinnamyl)-2-oxolycorine (S9)

[0133] S9 structure:

[0134] In a 250 mL round-bottom reaction flask, INE (5.0 mmol) and 4-bromocinnamic acid (6.0 mmol) were dissolved in 50 mL of anhydrous dichloromethane. Under nitrogen protection at room temperature, EDCI·HCl (9.0 mmol) and DMAP (0.5 mmol) were added sequentially. After the addition was complete, the reaction mixture was stirred and kept at 35 °C. HPLC monitoring showed that the remaining INE was less than 5%, at which point further processing was possible. The reaction solution was washed once with 100 mL of purified water and once with 100 mL of 10% sodium chloride solution, and concentrated under reduced pressure by column chromatography to obtain a white solid, S9. 1H NMR (600MHz, CDCl3) δ7.56(d,J=16.0Hz,1H),7.48(s,1H),7.46(s,1H),7.31(s,1H),7.30(s ,1H),6.79(s,1H),6.57(s,1H),6.26(d,J=16.0Hz,1H),6.13(d,J=3.1Hz,1H),6.02(s,1H), 5.90(s,1H),5.89(s,1H),4.19(d,J=14.1Hz,1H),3.63(d,J=14.0Hz,1H),3.48(dt,J=8.9,4 .6Hz,1H),3.33(d,J=10.1Hz,1H),3.25(d,J=9.9Hz,1H),2.89(s,2H),2.56(q,J=8.7Hz,1H); 13 C NMR (150MHz, CDCl3) δ192.85,169.03,165.33,146.74,146.69,144.65,133.08,132.09,129.52,128.8 1,125.16,124.76,120.49,117.76,107.32,105.48,101.10,69.15,62.40,56.32,53.26,45.61,30.01.

[0135] Example 14 1-(3-nitrocinnamyl)-2-oxolycorine (S10)

[0136] S10 structure:

[0137] In a 250 ml round-bottom reaction flask, INE (5.0 mmol) and 3-nitrocinnamic acid (6.0 mmol) were dissolved in 50 ml of anhydrous dichloromethane. Under nitrogen protection at room temperature, EDCI·HCl (9.0 mmol) and DMAP (0.5 mmol) were added sequentially. After the addition was complete, the reaction mixture was stirred and kept at 35 °C. HPLC monitoring showed that the remaining INE was less than 5%, at which point further processing was possible. The reaction solution was washed once with 100 ml of purified water and once with 100 ml of 10% sodium chloride solution, and concentrated under reduced pressure by column chromatography to obtain a white solid, S10. 1H NMR (400MHz, CDCl3) δ8.31(s,1H),8.20(dd,J=8.2,1.3Hz,1H),7.75(d,J=7.7Hz,1H),7.67(d,J=16 .0Hz,1H),7.55(t,J=8.0Hz,1H),6.78(s,1H),6.58(s,1H),6.42(d,J=16.0Hz,1H),6.14(d,J=2.8H z,1H),6.04(d,J=1.3Hz,1H),5.91(s,1H),5.90(s,1H),4.20(d,J=14.2Hz,1H),3.69–3.61(m,1H), 3.49(dd,J=8.4,4.1Hz,1H),3.36(d,J=9.8Hz,1H),3.30(s,1H),2.91(s,2H),2.60(d,J=7.2Hz,1H); 13 C NMR (100MHz, CDCl3) δ192.60,169.14,164.77,148.58,146.79,143.07,135.87,133.96,129.97,128.81,12 5.04,124.69,122.26,120.50,120.27,107.41,105.35,101.13,69.42,62.36,56.25,53.26,45.51,30.04.

[0138] Example 15 1-(2-chloro-5-nitrocinnamyl)-2-oxolycorine (S11)

[0139] S11 structure:

[0140] In a 250 mL round-bottom reaction flask, INE (5.0 mmol) and 2-chloro-5-nitrocinnamic acid (6.0 mmol) were dissolved in 50 mL of anhydrous dichloromethane. Under nitrogen protection at room temperature, EDCI·HCl (9.0 mmol) and DMAP (0.5 mmol) were added sequentially. After the addition was complete, the reaction mixture was stirred and kept at 35 °C. HPLC monitoring showed that the remaining INE content was less than 5%, at which point further processing was possible. The reaction solution was washed once with 100 mL of purified water and once with 100 mL of 10% sodium chloride solution, followed by concentrated solution under reduced pressure and column chromatography to obtain a white solid, S11. 1H NMR (600MHz, CDCl3) δ8.39(d,J=2.5Hz,1H),8.13(dd,J=8.8,2.6Hz,1H),8.03(d,J=15.9Hz,1H),7. 58(d,J=8.8Hz,1H),6.78(s,1H),6.59(s,1H),6.43(d,J=15.9Hz,1H),6.14(d,J=3.1Hz,1H),6.04(d ,J=1.6Hz,1H),5.92(s,1H),5.91(s,1H),4.20(d,J=14.1Hz,1H),3.65(d,J=14.1Hz,1H),3.52–3.44 (m,1H),3.36(d,J=10.0Hz,1H),3.28(d,J=9.8Hz,1H),2.90(d,J=6.9Hz,2H),2.57(q,J=8.7Hz,1H); 13 C NMR (150MHz, CDCl3) δ192.52,169.27,164.42,146.81,146.74,146.66,141.32,139.46,133.96,131.26,128.9 4,125.16,124.99,122.57,122.47,120.44,107.44,105.32,101.14,69.69,62.41,56.35,53.28,45.60,30.04.

[0141] Example 16 1-(2-nitro-5-bromocinnamyl)-2-oxolycorine (S12)

[0142] S12 structure:

[0143] In a 250 ml round-bottom reaction flask, INE (5.0 mmol) and 2-nitro-5-bromocinnamic acid (6.0 mmol) were dissolved in 50 ml of anhydrous dichloromethane. Under nitrogen protection at room temperature, EDCI·HCl (9.0 mmol) and DMAP (0.5 mmol) were added sequentially. After the addition was complete, the reaction mixture was stirred and kept at 35 °C. HPLC monitoring showed that the remaining INE was less than 5%, at which point further processing was possible. The reaction solution was washed once with 100 ml of purified water and once with 100 ml of 10% sodium chloride solution, and concentrated under reduced pressure by column chromatography to obtain a white solid, S12. 1H NMR (600MHz, CDCl3) δ8.00(d,J=15.8Hz,1H),7.91(d,J=8.7Hz,1H),7.69(d,J=2.0Hz,1H),7.63(dd,J =8.7,2.0Hz,1H),6.78(s,1H),6.57(s,1H),6.23(d,J=15.8Hz,1H),6.12(d,J=3.1Hz,1H),6.03(d,J= 1.8Hz,1H),5.92(s,2H),4.18(d,J=14.1Hz,1H),3.63(d,J=14.1Hz,1H),3.48(ddd,J=9.0,5.6,3.4Hz ,1H),3.35(d,J=9.9Hz,1H),3.24(d,J=9.4Hz,1H),2.90(dd,J=4.9,3.2Hz,2H),2.56(q,J=8.7Hz,1H); 13 C NMR (150MHz, CDCl3) δ192.49,169.24,163.95,146.81,146.79,14,6.75,139.79,133.33,132.13,132.00,128.8 9,128.41,126.48,124.92,123.30,120.44,107.36,105.40,101.15,69.74,62.45,56.35,53.26,45.68,30.03.

[0144] Example 17 : A new compound, 1-(2,4-dinitrocinnamyl)-2-oxolimoline (S13)

[0145] S13 structure:

[0146] In a 250 mL round-bottom reaction flask, INE (5.0 mmol) and 2,4-dinitrocinnamic acid (6.0 mmol) were dissolved in 50 mL of anhydrous dichloromethane. Under nitrogen protection at room temperature, EDCI·HCl (9.0 mmol) and DMAP (0.5 mmol) were added sequentially. After the addition was complete, the reaction mixture was stirred and kept at 35 °C. HPLC monitoring showed that the remaining INE content was less than 5%, at which point further processing was possible. The reaction solution was washed once with 100 mL of purified water and once with 100 mL of 10% sodium chloride solution, followed by concentrated solution and column chromatography under reduced pressure to obtain a white solid, S13. 1H NMR (600MHz, CDCl3) δ8.77(d,J=1.8Hz,1H),8.42(dd,J=8.6,1.7Hz,1H),7.91(d,J=8.7Hz,1H),6.71 (s,1H),6.55(s,1H),6.02(d,J=2.9Hz,1H),5.99(s,1H),5.95(s,1H),5.92(s,1H),5.24(dd,J=7.8, 3.8Hz,1H),4.15(d,J=14.0Hz,1H),3.55(d,J=14.0Hz,1H),3.46(t,J=5.9Hz,1H),3.27(d,J=9.6Hz, 1H),3.22–3.19(m,1H),3.12(d,J=10.1Hz,1H),2.87(d,J=7.4Hz,2H),2.51(dd,J=17.2,8.6Hz,1H); 13 C NMR (150MHz, CDCl3) δ192.37,169.13,168.24,148.06,147.26,146.77,146.70,144.21,130.22,128.89,12 7.58,125.01,120.40,120.12,107.31,105.43,101.18,69.49,62.36,56.32,53.25,45.54,29.99(s,-1H).

[0147] Example 18 1-(4-Methoxycinnamoyl)-2-oxolycorine (S14)

[0148] S14 structure:

[0149] In a 250 mL round-bottom reaction flask, INE (5.0 mmol) and 4-methoxycinnamic acid (6.0 mmol) were dissolved in 50 mL of anhydrous dichloromethane. Under nitrogen protection at room temperature, EDCI·HCl (9.0 mmol) and DMAP (0.5 mmol) were added sequentially. After the addition was complete, the reaction mixture was stirred and kept at 35 °C. HPLC monitoring showed that the remaining INE content was less than 5%, at which point further processing was possible. The reaction solution was washed once with 100 mL of purified water and once with 100 mL of 10% sodium chloride solution, followed by concentrated solution and column chromatography under reduced pressure to obtain a white solid, S14. 1H NMR (600MHz, CDCl3) δ7.59 (d, J = 15.9 Hz, 1H), 7.41 (s, 1H), 7.39 (s, 1H), 6.86 (s, 1H), 6.84 (s, 1H) ),6.81(s,1H),6.56(s,1H),6.14(d,J=16.0Hz,2H),6.02(d,J=1.6Hz,1H),5.90(d,J=1.1Hz,1H) ,5.89(s,1H),4.18(d,J=14.1Hz,1H),3.81(s,3H),3.63(d,J=14.0Hz,1H),3.51–3.45(m,1H),3 .32(d,J=10.0Hz,1H),3.26(d,J=9.9Hz,1H),2.89(dd,J=4.9,3.2Hz,2H),2.56(q,J=8.7Hz,1H); 13 C NMR (150MHz, CDCl3) δ193.12,168.91,165.88,161.52,146.70,146.67,145.74,129.87,128.77,126.94,125. 31,120.52,114.46,114.27,107.26,105.58,101.07,68.84,62.45,56.36,55.36,53.28,45.68,30.00(s,1H).

[0150] Example 19 : 1-(3,4-dioxane-cinnamoyl)-2-oxolimoline (S15)

[0151] S15 structure:

[0152] In a 250 mL round-bottom reaction flask, INE (5.0 mmol) and 3,4-dioxane-cinnamic acid (6.0 mmol) were dissolved in 50 mL of anhydrous dichloromethane. Under nitrogen protection at room temperature, EDCI·HCl (9.0 mmol) and DMAP (0.5 mmol) were added sequentially. After the addition was complete, the reaction mixture was stirred and kept at 35 °C. HPLC monitoring showed that the remaining INE content was less than 5%, at which point further processing was possible. The reaction solution was washed once with 100 mL of purified water and once with 100 mL of 10% sodium chloride solution, followed by concentrated solution under reduced pressure and column chromatography to obtain a white solid, S15. 1H NMR (400MHz, CDCl3) δ7.54(d,J=15.9Hz,1H),6.94(dd,J=4.0,2.6Hz,2H),6.80(s,1H),6.76(d, J=8.4Hz,1H),6.56(s,1H),6.13(d,J=3.1Hz,1H),6.10(d,J=15.9Hz,1H),6.02(d,J=1.6Hz,1H) ,5.98(s,2H),5.90(s,1H),5.89(s,1H),4.18(d,J=14.1Hz,1H),3.63(d,J=14.0Hz,1H),3.52–3 .42(m,1H),3.32(d,J=10.2Hz,1H),3.25(d,J=10.0Hz,1H),2.88(s,2H),2.56(q,J=8.6Hz,1H); 13 C NMR (100MHz, CDCl3) δ193.05,168.96,165.73,149.74,148.27,146.67,146.64,145.76,128.74,128.58,125.23,124.7 2,120.49,114.86,108.48,107.27,106.43,105.51,101.56,101.07,68.87,62.40,56.31,53.25,45.60,29.99(s,1H).

[0153] Example 20 1-Cinnamyl-2-oxolimanine (S16)

[0154] S16 structure:

[0155] In a 250 mL round-bottom reaction flask, INE (5.0 mmol) and cinnamic acid (6.0 mmol) were dissolved in 50 mL of anhydrous dichloromethane. Under nitrogen protection at room temperature, EDCI·HCl (9.0 mmol) and DMAP (0.5 mmol) were added sequentially. After the addition was complete, the reaction mixture was stirred and kept at 35 °C. HPLC monitoring showed that the remaining INE content was less than 5%, at which point further processing was possible. The reaction solution was washed once with 100 mL of purified water and once with 100 mL of 10% sodium chloride solution, followed by concentrated solution and column chromatography under reduced pressure to obtain a white solid, S16. 1H NMR(600MHz, CDCl3)δ7.64(d,J=16.0Hz,1H),7.46–7.42(m,2H),7.34(t,J=5.7Hz,3H),6 .80(s,1H),6.56(s,1H),6.28(d,J=16.0Hz,1H),6.14(d,J=3.1Hz,1H),6.03(s,1H),5.90 (s,1H),5.89(s,1H),4.19(d,J=14.1Hz,1H),3.63(d,J=14.1Hz,1H),3.50–3.46(m,1H), 3.33(d,J=10.0Hz,1H),3.27(d,J=10.0Hz,1H),2.93–2.83(m,2H),2.56(q,J=8.7Hz,1H); 13 C NMR (150MHz, CDCl3) δ192.96,168.94,165.56,146.73,146.70,146.07,134.17,130.47,128.84,128.76,1 28.16,125.21,120.53,117.04,107.30,105.52,101.09,69.03,62.43,56.32,53.26,45.63,30.01(s,1H).

[0156] Experimental example: Compound antiviral activity test

[0157] 1. Experimental Objective: This invention uses type I herpes simplex virus (HSV-1) for testing to evaluate the antiviral activity of the tested compounds. The virus strain tested was HSV-1GHSV-UL46.

[0158] 2. Test materials:

[0159] 2.1 Test Compound

[0160] The test compounds S1 to S17 (where S17 is INE) and lycorine of this invention are both solid powders, prepared into a 25 mM stock solution using 100% DMSO. In the first round of testing, a single concentration point was measured at 25 μM using double-duplicate wells. In the second round, eight concentration points were tested, with a 3-fold dilution and double-duplicate wells. The initial test concentration of the test compounds was 100 μM.

[0161] 2.2 Main Reagents and Instruments

[0162] (1) Main instruments: Microplate reader (Molecule Device, models SpectraMax340PC384 and SpectraMax340 iD3).

[0163] (2) Main reagents: CellTiter-Glo (Promega, catalog number G7573) and CCK-8 (Li Ji, AC11L057).

[0164] 3. Experimental Methods

[0165] The antiviral activity of the compounds was determined using the post-viral infection cytopathic effect (CPE) assay. The viral assay methods are summarized in Table 1.

[0166] Table 1. Virus Experimental Methods

[0167]

[0168]

[0169] Cells were seeded into 96-well cell culture plates at the cell density shown in Table 1 and cultured overnight in a 5% CO2, 37°C incubator. The next day, the test compound and virus were added. Cells were cultured for another 5 days in a 5% CO2, 33°C or 37°C incubator until obvious cytopathic effects were observed in the virus-infected control wells without the compound. Cell viability was then assessed using CellTiter-Glo or CCK-8 assays. If the cell viability in the wells containing the compound was higher than that in the virus-infected control wells, indicating a weakened cytopathic effect (CPE), it indicated that the compound inhibited the tested virus. The cytotoxicity assay was performed identically to the corresponding antiviral assay, but without viral infection.

[0170] The antiviral activity and cytotoxicity of the compounds were expressed as the inhibition rate (%) and cell viability (%) of the compounds against the virus-induced cellular viral effects, respectively. The calculation formulas are as follows:

[0171] Inhibition rate (%) = (Test well reading - Average value of virus control) / (Average value of cell control - Average value of virus control) × 100%;

[0172] Cell viability (%) = (Test well reading - Average value of culture medium control) / (Average value of cell control - Average value of culture medium control) × 100%;

[0173] EC 50 and CC 50 The values ​​were calculated using Prism software, and the inhibition curve fitting method was log(inhibitor) vs. response -- Variable slope of GraphPad Prism.

[0174] 4. Activity test results

[0175] The corresponding viral inhibitor was used as a positive control in the antiviral experiment.

[0176] In the first round of single-concentration point tests, all tested compounds showed superior inhibitory effects compared to lycorine. Eleven of the tested compounds (compounds S1, S2, S3, S5, S6, S7, S8, S9, S10, S11, and S12) showed inhibition rates greater than 50% against HSV-1 at a test concentration of 25 μM. The antiviral activity and cytotoxicity results of the tested and control compounds are shown in Table 2.

[0177] In the second round of antiviral testing, all tested compounds showed relatively higher CC values. 50 Values, particularly compounds S1, S2, S3, S5, S6, S7, S8, S9, S10, S11, and S12, showed antiviral activity against HSV-1 virus. Compound EC 50 The range is from 4.71 μM to 18.89 μM, CC 50 All were above 59.05 μM, and most compounds had Cd / C ratios of [missing value]. 50 All concentrations were above 100 μM, demonstrating good safety. Antiviral activity and cytotoxicity results are shown in Table 3.

[0178] Table 2. Results of HSV-1 virus inhibition rate and cell viability of the tested compounds in the first round of testing.

[0179]

[0180]

[0181] Table 3. Results of the second round of testing of the compounds' anti-HSV-1 virus activity

[0182]

[0183]

[0184] Note: CC 50 The median toxicity concentration (MCC) refers to the drug concentration at which 50% of the host cells are destroyed in an in vitro test. 50 The half-maximal effective concentration (MCI) refers to the concentration of a drug, antibody, or toxin that achieves 50% of its maximum biological effect. 50 and EC 50 It can be used to evaluate the safety and efficacy of drugs or substances. Generally speaking, CC... 50 The higher the EC 50 The lower the value, the safer and more effective the drug or substance.

[0185] Analysis of the data in the table above shows that the 1-substituted cinnamyl-2-oxolycorine compounds of this invention, represented by general formula I, exhibit varying degrees of inhibitory activity against type I herpes simplex virus (HSV-1) GHSV-UL46. Furthermore, their virus inhibition rates are superior to lycorine, and even superior to lycorine derivatives where the 2-hydroxyl group is oxidized while the 1-hydroxyl group is not esterified. More encouragingly, these compounds show significantly better toxicity than lycorine and higher safety, demonstrating promising potential for anti-HSV-1 drug use.

[0186] The above description is merely a preferred embodiment of this application and is not intended to limit this application. Although this application has been described in detail with reference to the foregoing embodiments, those skilled in the art can still modify the technical solutions described in the foregoing embodiments or make equivalent substitutions for some of the technical features. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of this application should be included within the protection scope of this application.

Claims

1. A 1-substituted cinnamyl-2-oxolimanesine compound or a pharmaceutically acceptable salt thereof, characterized in that, The compound is any one of the following: S1: 1-(2-fluorocinnamoyl)-2-oxo-l,2-dihydropenicillamine; S2: 1-(4-fluorocinnamoyl)-2-oxo-l,2-dihydropenicillamine; S3: 1-(3,4-difluorocinnamoyl)-2-oxo-l,2-dihydropenicillamine; S5: 1-(4-trifluoromethylcinnamoyl)-2-oxo-l,2-dihydropenicillamine; S6: 1-(3-trifluoromethoxycinnamoyl)-2-oxo-l,2-dihydropenicillamine; S7: 1-(2,5-dichlorocinnamoyl)-2-oxo-l,2-dihydropenicillamine; S8: 1-(2,6-dichlorocinnamoyl)-2-oxo-l,2-dihydropenicillamine; S9: 1-(4-bromocinnamoyl)-2-oxo-l,2-dihydropenicillamine; S10: 1-(3-nitrocinnamoyl)-2-oxo-l,2-dihydropenicillamine; S11: 1-(2-chloro-5-nitrocinnamoyl)-2-oxo-l,2-dihydropenicillamine; S12: 1-(2-nitro-5-bromocinnamoyl)-2-oxo-l,2-dihydropenicillamine.

2. A method for preparing the compound of claim 1, comprising: acetylating the hydroxyl groups at positions 1 and 2 of penicillamine using acetic anhydride to obtain an intermediate INB, taking penicillamine as a starting compound; selectively removing the acetyl group at position 2 of the intermediate INB under acidic conditions to obtain an intermediate INC; oxidizing the hydroxyl group at position 2 of the intermediate INC to obtain an intermediate IND; removing the acetyl group at position 1 of the intermediate IND under acidic conditions to obtain an intermediate INE; The intermediate INE is subjected to an esterification reaction with a substituted cinnamic acid in the presence of a condensing agent to obtain the compound of claim 1 ; the substituted cinnamic acid is selected from the group consisting of: 2 fluorocinnamic acid, 4 fluorocinnamic acid, 3,4 difluorocinnamic acid, 4 trifluoromethylcinnamic acid, 3 trifluoromethoxycinnamic acid, 2,5 dichlorocinnamic acid, 2,6 dichlorocinnamic acid, 4 bromocinnamic acid, 3 nitrocinnamic acid, 2 chloro 5 nitrocinnamic acid, 2 nitro 5 bromocinnamic acid; 。 3. A pharmaceutical composition comprising the compound of claim 1 or a pharmaceutically acceptable salt thereof.

4. A pharmaceutical preparation comprising the compound of claim 1 or a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable adjuvant or pharmaceutical carrier.

5. Use of a compound of claim 1 or a pharmaceutically acceptable salt thereof or a pharmaceutical composition of claim 3 or a pharmaceutical preparation of claim 4 in the manufacture of an antiviral medicament, wherein, The virus is herpes simplex virus type I.

6. Use of the compound of claim 1 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition of claim 3 or the pharmaceutical preparation of claim 4 in the preparation of a drug for treating a disease associated with herpes simplex virus type I infection.

Images