Isoquinoline drugs and their use in improving or treating fungal infections

Abstract

The present application relates to the technical field of small molecule drugs, and particularly relates to an isoquinoline drug and application thereof in improving or treating fungal infection. The isoquinoline drug has good bacteriostatic activity on Candida albicans. Further research and observation show that the compound has inhibitory effect on adhesion, mycelium formation, biofilm formation and pathogenicity of Candida albicans, which indicates that the 10,13-substituted berberine derivative can be used for preventing and / or treating fungal infection, in particular, Candida albicans infection.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of small molecule drugs, and particularly relates to an isoquinoline drug and application thereof in improving or treating fungal infection. BACKGROUND

[0002] In recent years, the continuous expansion of malignant tumor patients and HIV infected population, combined with the widespread use of immunosuppressive drugs, leads to a significant growth trend of invasive fungal infection. Among the numerous opportunistic pathogenic fungi, Candida, especially Candida albicans (CA), has become the pathogen with the highest detection rate in clinic. It is worth noting that CA not only has strong tissue invasion ability, but also has an increasingly serious drug resistance trend, which makes the deep infection caused by the fungus present a high mortality rate, which poses a new challenge to clinical antifungal treatment.

[0003] The current clinical first-line antifungal drugs mainly include three categories: azoles (fluconazole, etc.), polyenes (amphotericin B, etc.) and echinocandins (caspofungin, etc.). However, the frequent occurrence of CA drug-resistant mutant strains caused by long-term drug exposure significantly weakens the clinical efficacy of these traditional drugs. In addition, the above-mentioned drugs generally have treatment limitations, which can not only induce gastrointestinal reactions, allergic syndromes, liver and kidney damage and other adverse reactions, but also have nervous system side effects such as headache and fever, which seriously affect the treatment compliance of patients.

[0004] In view of this clinical dilemma, the development of antifungal preparations with new mechanisms of action has become a key breakthrough to solve the problem of drug resistance. SUMMARY

[0005] The present application aims to at least solve one of the problems in the related art. To this end, one object of the present application is to provide a compound, a product comprising the compound and use thereof, which has good antibacterial activity against Candida albicans, and further research and observation of the inhibitory effect of the compound on the adhesion, mycelium formation, biofilm formation and pathogenicity of Candida albicans show that the 10,13-substituted berberine derivative of the present application can be used for preventing and / or treating fungal infection, especially Candida albicans infection.

[0006] To this end, the first aspect of the present application provides a compound, which is a compound represented by general formula (I) or a pharmaceutically acceptable salt of the compound represented by general formula (I):

[0007]

[0008] R1, R2 are each independently selected from - (CH2) m R3, - (CH2) n -CN, C2-C10 alkenyl, unsubstituted C2-C 10 alkynyl, unsubstituted or substituted by at least one R a substituted C1-C 10 alkyl;

[0009] m = 1, n = 1-5,

[0010] R3is unsubstituted or substituted by at least one R b substituted phenyl;

[0011] R a is -NH2;

[0012] R b is selected from -NO2, halogen, C1alkoxy, C1alkyl,

[0013] and R1, R2are not simultaneously unsubstituted C1-C 10 alkyl.

[0014] The current clinical antifungal treatment system mainly relies on four categories of drugs: azoles, polyenes, echinocandins and flucytosine. However, the application of drugs has significant limitations: although polyene drugs such as nystatin have strong killing effect, they have low bioavailability (<5%) and obvious nephrotoxicity; and the problem of drug resistance caused by long-term antifungal treatment is becoming increasingly serious, especially the dynamic regulation mechanism of virulence factors such as biofilm formation and hyphal differentiation significantly enhances the tissue invasion and drug resistance of pathogens. These factors together lead to the clinical dilemma of decreasing efficacy of existing treatment regimens.

[0015] In recent years, it has been found that the traditional Chinese medicine active ingredient berberine (BBR) exhibits unique antifungal properties. Its mechanism of action involves: 1) inhibition of hyphal differentiation and biofilm formation; 2) up-regulation of cell wall β-1, 3-glucan and chitin synthesis-related gene expression; 3) interference with the quorum sensing system to reduce the secretion of virulence factors. However, due to the high minimum inhibitory concentration (MIC = 16 μg / ml) and poor pharmacokinetic properties, BBR has not yet reached the clinical conversion standard.

[0016] Based on the above background, the present research focuses on the structural optimization of berberine, and through computer-aided drug design and structure-activity relationship analysis, the key pharmacophore groups such as C-10, 13-hydroxyl and C-13-methoxyl of the mother nucleus are modified. The target innovative derivatives have the following advantages: 1) the antibacterial activity is increased by 5-10 times compared with the parent; 2) breakthrough existing drug resistance mechanism; 3) improve solubility and tissue permeability; 4) reduce host cell toxicity. Finally, the inventors screened and obtained compounds represented by structural formula (I), and found that the compounds have good antibacterial activity against Candida albicans. Further research and observation of the compounds on the adhesion, hypha formation, biofilm formation and pathogenicity of Candida albicans showed that the 10, 13-substituted berberine derivatives of the present application can be used for preventing and / or treating fungal infections, especially Candida albicans infections. Therefore, the 10, 13-substituted berberine derivatives of the present application can be used for preparing drugs for preventing and / or treating infectious diseases caused by Candida albicans.

[0017] According to embodiments of the present application, the compound is selected from one of the following structures:

[0018]

[0019]

[0020]

[0021]

[0022] The second aspect of the present application provides a medicine. According to embodiments of the present application, the medicine comprises the compound of the first aspect, and the medicine is used for improving or treating Candida albicans infection.

[0023] According to embodiments of the present application, the medicine further comprises a pharmaceutically acceptable adjuvant, such as a pharmaceutically acceptable carrier, diluent or excipient.

[0024] The medicine prepared by using the compound provided by the present application in various dosage forms is administered to a subject in a therapeutically effective amount, and after being absorbed by the subject, the medicine can treat or improve fungal infection.

[0025] According to embodiments of the present application, the medicine further comprises one or more other therapeutic agents. Among them, the other therapeutic agents are similar in function to the compounds of the present application, and can be used for treating or improving fungal infection.

[0026] The present application relates to the compound of general formula (I) or (I) suitable pharmaceutically acceptable salt, including but not limited to hydrochloride, hydrobromic acid, sulfuric acid or hydrogen sulfate, phosphate or hydrogen phosphate, acetate, benzoate, succinate, fumarate, maleate, lactate, citrate, tartrate, gluconate, methanesulfonate, benzenesulfonate or p-toluenesulfonate. According to the foregoing, any of the compounds mentioned herein includes its pharmaceutically acceptable salt, solvate or combination thereof.

[0027] In addition to the pharmaceutically acceptable salts of the compounds of the present application, other salts are included within the scope of the present application. These can be formed by salts during the purification of the compounds or in the preparation of other pharmaceutically acceptable salts or can be used in the identification, characterization or purification of the compounds of the present application.

[0028] The third aspect of the present application provides the use of the compound of the first aspect in the preparation of a medicament. According to the embodiments of the present application, the medicament is used to improve or treat Candida albicans infection.

[0029] The anti-fungal compound 10,13-substituted berberine derivative developed in the present study exhibits multiple advantages: it does not mainly kill Candida albicans, but plays a role by inhibiting the adhesion and pathogenicity of Candida albicans; the 10,13-substituted berberine derivative of the present application is expected to be used in combination with other first-line anti-fungal drugs, thereby producing a synergistic effect against drug-resistant Candida albicans.

[0030] The anti-fungal compound 10,13-substituted berberine derivative of the present application can target the inhibition of CA biofilm formation. More attention is paid to the fact that the compound exhibits excellent safety characteristics while exerting significant anti-fungal activity, and has no significant toxic effect on host cells. These characteristics make it have important conversion value in the field of anti-CA infection drug research and development, and provide a new solution for dealing with fungal infections.

[0031] Additional aspects and advantages of the present application will be partially given in the following description, partially will become obvious from the following description, or will be understood by the practice of the present application. BRIEF DESCRIPTION OF DRAWINGS

[0032] The above and / or additional aspects and advantages of the present application will become apparent and more readily appreciated from the following description, taken in conjunction with the following drawings, of embodiments of the present application, wherein:

[0033] Figure 1 The effect of BBR and compound 23 on the thickness of Candida albicans biofilm is shown by CLSM detection;

[0034] Figure 2 The effect of BBR or compound 23 on the tongue mucosa of oropharyngeal candidiasis mice is shown;

[0035] Figure 3 Shown are the effects of BBR or compound 23 treatment on the oral fungal load in mice with oropharyngeal candidiasis;

[0036] Figure 4 shows the results of HE staining of tongue tissues of mice with oropharyngeal candidiasis after treatment with BBR or compound 23;

[0037] Figure 5 Shown are the effects of drugs on the tongue mucosal surface of mice with oropharyngeal candidiasis after treatment with BBR or compound 23, as observed by SEM;

[0038] Figure 6 The results show that BBR or compound 23 can significantly affect the tongue tissue of mice with oropharyngeal candidiasis. S100a8 Effects on gene expression;

[0039] Figure 7 Shown are the effects of BBR or compound 23 treatment on EGFR protein expression in tongue tissues of mice with oropharyngeal candidiasis. DETAILED DESCRIPTION

[0040] The embodiments of the present invention are described in detail below. The embodiments described below are exemplary and are only used to explain the present invention, and should not be understood as limiting the present invention.

[0041] It should be noted that the terms "first" and "second" are used for descriptive purposes only and should not be understood to indicate or imply relative importance or implicitly specify the number of the technical features indicated. Therefore, features defined as "first" or "second" may explicitly or implicitly include one or more of such features. Furthermore, in the description of the present invention, unless otherwise specified, "plurality" means two or more.

[0042] The endpoints of the ranges and any values ​​disclosed herein are not limited to the precise ranges or values, and these ranges or values ​​should be understood to include values ​​close to these ranges or values. For numerical ranges, the endpoints of each range, the endpoints of each range and individual point values, and the individual point values ​​can be combined with each other to obtain one or more new numerical ranges, which should be considered to be specifically disclosed herein.

[0043] In order to make the present invention more easily understood, certain technical and scientific terms are specifically defined below. Unless otherwise clearly defined elsewhere in this document, all other technical and scientific terms used herein have the meaning commonly understood by those skilled in the art to which the present invention belongs.

[0044] In this text, the term "comprising" or "including" is an open term, which means that it includes the recited elements, but not excluding other elements.

[0045] In this text, the term "optionally", "optional" or "optional" generally means that the event or circumstance subsequently described can or can not occur, and that the description includes situations where the event or circumstance occurs, as well as situations where it does not.

[0046] Unless otherwise indicated, the definitions of groups and terms recited in the specification and claims of this application, including the definitions of examples, illustrative examples, preferred definitions, definitions recited in tables, definitions of specific compounds in examples, etc., can be combined and incorporated with each other in any manner. The group definitions and compound structures after such combination and incorporation should be within the scope recited in the specification of this application.

[0047] The term "pharmaceutically acceptable salt" refers to a non-toxic, pharmaceutically acceptable salt of an acid or a base, including salts of inorganic acids and bases, salts of organic acids and bases. Salts derived from inorganic bases include, but are not limited to, metal salts of Al, Ca, Li, Mg, K, Na and Zn; salts derived from organic bases include, but are not limited to, salts of primary, secondary or tertiary amines, including naturally occurring substituted or unsubstituted amines, cyclic amines and basic ion exchange resins, such as ammonium, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, diethanolamine, ethanolamine, dimethylethanolamine, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, caffeine, procaine, choline, betaine, phenylamine penicillin, ethylenediamine, glucosamine, methylglucosamine, theobromine, triethanolamine, tromethamine, purines, piperazine, piperidine, N-ethylpiperidine or polyamine resin; salts derived from inorganic acids and organic acids include, but are not limited to, organic salts of sulfuric acid, phosphoric acid, nitric acid, hydrobromic acid, hydrochloric acid, formic acid, acetic acid, propionic acid, benzenesulfonic acid, benzoic acid, phenylacetic acid, salicylic acid, alginic acid, anthranilic acid, camphoric acid, citric acid, ethenesulfonic acid, formic acid, fumaric acid, furoic acid, gluconic acid, glucuronic acid, glutamic acid, glycolic acid, isethionic acid, lactic acid, maleic acid, malic acid, mandelic acid, mucic acid, pamoic acid, pantothenic acid, stearic acid, succinic acid, sulfosalicylic acid, tartaric acid, p-toluenesulfonic acid, malonic acid, 2-hydroxypropanoic acid, oxalic acid, glycolic acid, galacturonic acid, citric acid, lysine, arginine, aspartic acid, cinnamic acid, methanesulfonic acid, ethanesulfonic acid or trifluoromethanesulfonic acid, etc.

[0048] The term "drug" means a mixture of one or more compounds described in the present application or physiologically / pharmaceutically acceptable salts or prodrugs thereof with other chemical components, such as physiologically / pharmaceutically acceptable carriers and excipients. The purpose of the drug is to facilitate the administration of the compound to the organism, to facilitate the absorption of the active ingredient and to exert biological activity.

[0049] The term "C1-C 10 The term "alkyl" is understood to mean a linear or branched, saturated, monovalent hydrocarbon radical having 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms. Such alkyl radicals are, for example, methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, 2-methylbutyl, 1-methylbutyl, 1-ethylpropyl, 1,2-dimethylpropyl, neopentyl, 1,1-dimethylpropyl, 4-methylbutyl, "C1-C6 alkyl" is understood to mean a straight-chain or branched saturated monovalent hydrocarbon group having 1, 2, 3, 4, 5 or 6 carbon atoms.

[0050] The term "excipient" refers to a pharmaceutically acceptable inert ingredient. Non-limiting examples of the term "excipient" include binders, disintegrants, lubricants, glidants, stabilizers, fillers, and diluents. Excipients enhance the handling characteristics of a pharmaceutical formulation, i.e., by increasing flowability and / or cohesiveness, making the formulation more suitable for direct compression. Examples of typical "pharmaceutically acceptable carriers" suitable for the above-mentioned preparations are: sugars, such as lactose, sucrose, mannitol and sorbitol, or corn starch, tapioca starch and potato starch; cellulose and its derivatives, such as sodium carboxymethylcellulose, ethyl cellulose and methyl cellulose; calcium phosphates, such as dicalcium phosphate and tricalcium phosphate; sodium sulfate; calcium sulfate; polyvinyl pyrrolidone; polyvinyl alcohol; stearic acid; alkaline earth metal stearates, such as magnesium stearate and calcium stearate; vegetable oils, such as peanut oil, cottonseed oil, sesame oil, olive oil and corn oil; nonionic, cationic and anionic surfactants; ethylene glycol polymers; fatty alcohols; and cereal hydrolyzed solids and other non-toxic compatible fillers, binders, disintegrants, buffers, preservatives, antioxidants, lubricants, colorants and the like excipients commonly used in pharmaceutical preparations.

[0051] Candida albicans, a resident commensal microorganism of the human body, is widely present in mucosal systems such as the skin and oral cavity, digestive tract, and genitourinary tract of healthy individuals. This pathogen often presents as asymptomatic colonization or superficial infection in immunocompetent individuals. However, in immunocompromised individuals (such as patients undergoing chemotherapy for hematologic malignancies, solid organ transplant recipients, and HIV / AIDS patients), it can cause systemic infection with a mortality rate of up to 40%. It is a major pathogen causing fungemia, disseminated infection, and intensive care unit-associated sepsis.

[0052] Based on this, the present invention aims to provide a class of 10,13-disubstituted berberine derivatives and their preparation methods and applications. Through the rational design and modification of BBR, candidate drugs with improved activity, novel mechanisms, low resistance to drug resistance, and good safety are obtained.

[0053] According to a specific embodiment of the present invention, the present invention provides a compound, which is a compound represented by general formula (I) or a pharmaceutically acceptable salt of the compound represented by general formula (I):

[0054]

[0055] R1 and R2 are each independently selected from -(CH2) m R3, -(CH2) n -CN, unsubstituted C2-C 10 Alkenyl, unsubstituted C2-C 10 Alkynyl, unsubstituted or substituted with at least one R a Substituted C1-C 10 alkyl;

[0056] m=1, n=1~5,

[0057] R3 is unsubstituted or substituted with at least one R b substituted phenyl;

[0058] R a is -NH2;

[0059] R b Selected from -NO2, halogen, C1 alkoxy, C1 alkyl,

[0060] and R1 and R2 are not unsubstituted C1-C 10 Alkyl. Halogen is fluorine, chlorine, bromine or iodine.

[0061] It should be noted that m is 1, and n can be 1, 2, 3, 4, or 5. For example, n is 1-3, 1-4, or 1-5.

[0062] According to a specific embodiment of the present invention, the present invention provides a method for preparing a 10,13-substituted berberine derivative, comprising the following steps:

[0063]

[0064] The (M-2), anhydrous acetonitrile, potassium carbonate and the first raw material are reacted at 60-70 ℃, the reaction progress is monitored by TLC, after the reaction is completed, it is cooled to solid completely precipitates, suction filtration, the filtrate is silica gel sample, dichloromethane and methanol are used as mobile phase, and the yellow intermediate (M-3) is obtained by Flash fast column chromatography purification.The above obtained intermediate is reacted with anhydrous acetonitrile, sodium hydrogen and the second raw material at 70-90 ℃, the reaction progress is monitored by TLC, after the reaction is completed, it is neutralized with hydrochloric acid methanol solution, and then silica gel sample is obtained, and the yellow end product is obtained by Flash fast column chromatography purification using dichloromethane and methanol as mobile phase.

[0065] According to a specific embodiment of the present application, the application provides the use of the aforementioned 10,13-position substituted berberine derivative or physiologically acceptable salt or pharmaceutical composition in the preparation, prevention and / or treatment of products of microbial infection.

[0066] Further, the microorganism is Candida albicans.

[0067] According to a specific embodiment of the present application, the application provides the use of the aforementioned 10,13-position substituted berberine derivative or physiologically acceptable salt or pharmaceutical composition in the preparation, prevention and / or treatment of antibacterial drugs.

[0068] The present application has the following beneficial effects:

[0069] The present application provides a kind of 10,13-position substituted berberine derivative and its preparation method and application, and in the early work, efficient, low toxicity, compound is screened for not easy to produce drug resistance. Compared with the minimum inhibitory concentration (MIC) of BBR, the 10,13-position substituted berberine derivative prepared by the method of the present application shows more promising antibacterial activity against Candida albicans. Then, Candida albicans is used as the test object, and the influence of 10,13-position substituted berberine derivative on Candida albicans filament and biofilm formation is investigated. The purpose is to further affect the invasion of Candida albicans by detecting the interference of 10,13-position substituted berberine derivative on the virulence formation factor of Candida albicans. The results show that 10,13-position substituted berberine derivative has good inhibitory effect on the formation of Candida albicans filament. Moreover, 10,13-position substituted berberine derivative itself has less toxicity, does not affect the normal growth of human cells and Candida albicans, and thus is not easy to produce drug resistance. This has good application prospect in the development of new antifungal drugs, especially in the development of drugs against Candida albicans infection.

[0070] Therefore, the application of the 10,13-position substituted berberine derivative in the preparation of a drug for resisting Candida albicans infection, and the application of the 10,13-position substituted berberine derivative in the preparation of a drug for preventing and / or treating an infectious disease caused by Candida albicans should be within the scope of protection of the present application.

[0071] The schemes of the present disclosure will be explained below in conjunction with examples. Those skilled in the art will understand that the examples below are only for illustration of the present disclosure and should not be regarded as limiting the scope of the present disclosure. If no specific technique or condition is specified in the examples, the technique or condition described in the literature in the art or according to the product instruction is used. If no manufacturer of the reagent or instrument is specified, it is a conventional product that can be obtained commercially.

[0072] Example 1 Synthesis of 13-((E)-but-2-en-1-yl)-10-(((E)-but-2-en-1-yl)oxy)-9-methoxy-5,6-dihydro-[1,3]dioxolo[4,5-g]isoquinoline[3,2-a]isoquinoline-7-ol (Compound 1)

[0073]

[0074] M-2, anhydrous acetonitrile, potassium carbonate and 3-bromopropene (first raw material) were reacted at 60-70 ℃, the reaction progress was monitored by TLC, after the reaction was completed, it was cooled to solid completely precipitated, suction filtration, the filtrate was silica gel sample, with dichloromethane and methanol as mobile phase, by Flash fast column chromatography to obtain yellow intermediate. The above obtained intermediate was reacted with anhydrous acetonitrile, sodium hydride and 3-bromopropene (second raw material) at 70-90 ℃, the reaction progress was monitored by TLC, after the reaction was completed, it was neutralized with hydrochloric acid methanol solution, then silica gel sample was mixed, and dichloromethane and methanol were used as mobile phase, and the yellow final product 1 was obtained by Flash fast column chromatography. The nuclear magnetic resonance result of the final product is as follows:

[0075] 1 H NMR (500 MHz, DMSO-d6) δ 9.85 (s, 1H), 7.47 (s, 1H), 7.31 (d, J = 8.8Hz, 1H), 7.17 (d, J = 8.8 Hz, 1H), 6.99 (t, J = 1.0 Hz, 1H), 6.11 – 5.99 (m, 1H),6.04 (s, 2H), 5.89 (tt, J = 16.4, 8.1 Hz, 1H), 5.39 (ddt, J= 16.8, 2.2, 1.0 Hz,1H), 5.31 (ddt, J = 16.8, 2.2, 1.0 Hz, 1H), 5.17 (ddt, J = 16.4, 2.1, 1.0 Hz,1H), 5.07 (ddt, J = 16.4, 2.1, 1.0 Hz, 1H), 4.67 – 4.58 (m, 4H), 3.40 (dt, J =8.1, 1.0 Hz, 2H), 3.16 (dddd, J = 19.3, 7.6, 4.6, 1.0 Hz, 2H).

[0076] 13 C NMR (125 MHz, DMSO-d6) δ 149.96, 148.41, 148.40, 147.87, 143.00,137.82, 136.41, 132.32, 130.42, 130.40, 129.28, 122.76, 122.53, 122.03,118.82, 118.76, 116.20, 109.29, 109.20, 101.61, 69.70, 62.21, 58.66, 34.75,28.27.

[0077] ESI + :402.47.

[0078] Example 2 Synthesis of 13-((E)-but-2-en-1-yl)-10-(((E)-but-2-en-1-yl)oxy)-9-methoxy-5,6-dihydro-[1,3]dioxolo[4,5-g]isoquinoline[3,2-a]isoquinoline-7-carboxylic acid (Compound 2)

[0079]

[0080] The experimental procedure was the same as Example 1, except that the first starting material was crotyl bromide and the second starting material was crotyl bromide; the final product NMR results are as follows:

[0081] 1 H NMR (500 MHz, DMSO-d6) δ 9.85 (s, 1H), 7.47 (s, 1H), 7.31 (d, J = 8.8Hz, 1H), 7.17 (d, J= 8.8 Hz, 1H), 6.99 (t, J = 1.1 Hz, 1H), 6.04 (s, 2H), 5.87 –5.78 (m, 1H), 5.73 – 5.58 (m, 3H), 4.69 (dt, J = 4.4, 1.0 Hz, 2H), 4.64 (ddd, J =7.3, 4.6, 1.7 Hz, 2H), 4.01 (s, 3H), 3.43 (dt, J = 7.1, 1.1 Hz, 2H), 3.16(dddd, J = 19.3, 7.6, 4.6, 1.0 Hz, 2H), 1.67 (ddt, J = 5.3, 1.9, 1.0 Hz, 3H),1.62 (dt, J = 5.3, 1.2 Hz, 3H).

[0082] 13 C NMR (125 MHz, DMSO-d6) δ 149.57, 148.46, 147.54, 147.51, 142.86,136.66, 130.42, 130.25, 128.85, 128.62, 128.26, 127.61, 127.45, 122.33,122.02, 121.57, 117.85, 108.80, 108.41, 101.51, 69.24, 61.65, 58.15, 33.10,27.89, 17.90, 17.70.

[0083] ESI + :430.52.

[0084] Example 3 Synthesis of 9-methoxy-13-(3-methylbut-2-en-1-yl)-10-((3-methylbut-2-en-1-yl)oxy)-5,6-dihydro-[1,3]dioxolo[4,5g]isoquinoline[3,2-a]isoquinoline-7-carboxylic acid (Compound 3)

[0085]

[0086] The experimental procedure was the same as Example 1, except that the first starting material was 3,3-dimethylallyl bromide and the second starting material was 3,3-dimethylallyl bromide; the final product NMR results were as follows:

[0087] 1 H NMR (500 MHz, DMSO-d6) δ 9.85 (s, 1H), 7.47 (s, 1H), 7.31 (d, J = 8.8Hz, 1H), 7.17 (d, J = 8.8 Hz, 1H), 6.99 (t, J = 1.1 Hz, 1H), 6.04 (s, 2H), 5.31(ddt, J = 7.3, 5.7, 1.6 Hz, 1H), 5.28 (ddt, J = 6.6, 3.2, 1.7 Hz, 1H), 4.67 –4.59 (m, 4H), 4.01 (s, 3H), 3.46 (dq, J = 7.4, 1.0 Hz, 2H), 3.16 (dddd, J = 19.3,7.6, 4.6, 1.0 Hz, 2H), 1.75 (q, J = 1.1 Hz, 6H), 1.68 (q, J = 1.2 Hz, 6H).

[0088] 13 C NMR (125 MHz, DMSO-d6) δ 149.57, 148.33, 147.54, 147.51, 142.84,137.69, 136.72, 130.25, 130.20, 129.75, 128.51, 123.77, 122.33, 122.02,121.52, 119.15, 117.85, 108.80, 108.41, 101.51, 66.11, 61.65, 58.15, 28.68,27.89, 24.63, 24.49, 19.90, 19.28.

[0089] ESI + :458.58.

[0090] Synthesis of 10-(3-aminopropoxy)-13-(3-aminopropyl)-9-methoxy-5,6-dihydro- [1,3]dioxolo[4,5-g]isoquinoline[3,2-a]isoquinoline-7-ium (Compound 4)

[0091]

[0092] The experimental procedure was the same as in Example 1, except that the first starting material was 1-bromopropylamine and the second starting material was 1-bromopropylamine; the final product NMR results were as follows:

[0093] 1 H NMR (500 MHz, DMSO-d6) δ 9.85 (s, 1H), 7.47 (s, 1H), 7.37 (d, J = 8.8Hz, 1H), 7.12 (d, J = 8.8 Hz, 1H), 6.99 (t, J = 1.0 Hz, 1H), 6.04 (s, 2H), 4.63(ddd, J = 7.6, 4.9, 1.2 Hz, 2H), 4.09 – 4.00 (m, 5H), 3.16 (dddd, J = 19.3, 7.6,4.6, 1.0 Hz, 2H), 2.97 – 2.89 (m, 2H), 2.87 – 2.78 (m, 6H), 2.02 (p, J = 5.6Hz, 2H), 1.87 (tt, J = 8.1, 5.3 Hz, 2H), 1.74 (t, J = 6.3 Hz, 2H).

[0094] 13 C NMR (125 MHz, DMSO-d6) δ 149.57, 148.33, 147.55, 147.54, 142.81,137.12, 134.45, 132.51, 132.39, 121.96, 121.92, 121.29, 118.04, 109.19,108.80, 101.51, 67.72, 61.65, 58.81, 40.83, 37.13, 31.06, 29.70, 29.09,27.89.

[0095] ESI + : 436.53.

[0096] Synthesis of 13-(hept-6-en-1-yl)-10-(hept-6-en-1-yloxy)-9-methoxy-5,6-dihydro- [1,3]dioxolo[4,5-g]isoquinoline[3,2-a]isoquinoline-7-ium (Compound 5)

[0097]

[0098] The experimental procedure was the same as in Example 1, except that the first starting material was 7-bromoheptene and the second starting material was 7-bromoheptene; the final product NMR results were as follows:

[0099] 1 H NMR (500 MHz, DMSO-d6) δ 9.85 (s, 0H), 7.37 (d, J = 8.8 Hz, 0H), 6.04(s, 1H), 5.77 (tt, J = 17.3, 6.9 Hz, 1H), 5.12 (ddt, J = 17.1, 2.1, 1.0 Hz, 1H),4.97 (ddt, J = 17.1, 2.2, 1.0 Hz, 1H), 4.63 (ddd, J = 7.6, 4.9, 1.2 Hz, 1H), 4.00(t, J = 6.1 Hz, 1H), 3.16 (dddd, J = 19.3, 7.6, 4.6, 1.0 Hz, 1H), 2.78 (t, J = 8.5Hz, 1H), 2.07 – 1.99 (m, 2H), 1.73 – 1.64 (m, 2H), 1.43 (qd, J = 6.7, 5.6 Hz,1H), 1.39 – 1.29 (m, 3H).

[0100] 13 C NMR (125 MHz, DMSO-d6) δ 149.57, 148.34, 147.55, 147.54, 142.81,139.08, 138.84, 137.36, 135.30, 134.45, 132.47, 121.95, 121.89, 121.29,118.04, 114.61, 114.41, 109.19, 108.80, 101.51, 68.79, 61.65, 58.81, 33.79,33.67, 30.56, 29.09, 28.72, 28.64, 28.49, 28.44, 27.89, 25.86.

[0101] ESI + :514.69.

[0102] Example 6 Synthesis of 9-methoxy-13-(pent-4-en-1-yl)-10-(pent-4-en-1-yloxy)-5,6- dihydro-[1,3]dioxolo[4,5-g]isoquinoline[3,2-a]isoquinoline-7-carboxylic acid (Compound 6)

[0103]

[0104] The experimental procedure was the same as Example 1, except that the first starting material was 5-bromopentene and the second starting material was 5-bromopentene; the final product NMR results are as follows:

[0105] 1 H NMR (500 MHz, DMSO-d6) δ 9.85 (s, 1H), 7.47 (s, 1H), 7.37 (d, J = 8.8Hz, 1H), 7.12 (d, J = 8.8 Hz, 1H), 6.99 (t, J = 1.0 Hz, 1H), 6.04 (s, 2H), 5.73(tdt, J = 17.3, 10.5, 6.8 Hz, 2H), 5.12 (dddt, J = 17.1, 5.1, 2.1, 1.0 Hz, 2H),4.97 (dtt, J = 17.1, 2.2, 1.0 Hz, 2H), 4.63 (ddd, J = 7.6, 4.9, 1.2 Hz, 2H), 4.06(t, J = 6.8 Hz, 2H), 4.02 (s, 3H), 3.16 (dddd, J = 19.3, 7.6, 4.6, 1.0 Hz, 2H),2.88 (s, 0H), 2.85 (s, 0H), 2.18 – 2.09 (m, 4H), 1.77 (tt, J = 8.4, 6.8 Hz,2H), 1.65 (tt, J = 9.3, 7.9 Hz, 2H).

[0106] 13C NMR (125 MHz, DMSO-d6) δ 149.57, 148.33, 147.55, 147.54, 142.81, 138.25, 137.61, 137.10, 134.45, 133.11, 132.48, 121.95, 121.91, 121.29, 118.04, 115.14, 114.91, 109.19, 108.80, 101.51, 69.02, 61.66, 58.81, 33.02, 30.17, 30.13, 28.67, 27.89, 27.85.

[0107] ESI + :458.58.

[0108] Example 7 Synthesis of 9-methoxy-13-(oct-7-yn-1-yl)-10-(oct-7-yn-1-yloxy)-5,6-dihydro- [1,3]dioxolo[4,5-g]isoquinoline[3,2-a]isoquinoline (Compound 7)

[0109]

[0110] The experimental procedure was the same as Example 1, except that the first starting material was 8-bromooct-7-yne and the second starting material was 8-bromooct-7-yne; the final product NMR results are as follows:

[0111] 1 H NMR (500 MHz, DMSO-d6) δ 9.85 (s, 0H), 7.37 (d, J = 8.8 Hz, 0H), 6.04(s, 1H), 4.63 (ddd, J = 7.6, 4.9, 1.2 Hz, 1H), 4.00 (t, J = 6.1 Hz, 1H), 3.16(dddd, J = 19.3, 7.6, 4.6, 1.0 Hz, 1H), 2.78 (t, J = 8.5 Hz, 1H), 2.12 (td, J =5.9, 3.0 Hz, 2H), 2.06 (t, J = 2.9 Hz, 1H), 1.77 (tt, J = 7.5, 6.1 Hz, 1H), 1.68(tt, J= 8.4, 7.2 Hz, 1H), 1.55 – 1.33 (m, 6H).

[0112] 13 C NMR (125 MHz, DMSO-d6) δ 149.57, 148.34, 147.55, 147.54, 142.81,137.36, 135.30, 134.45, 132.47, 121.95, 121.89, 121.29, 118.04, 109.19,108.80, 101.51, 83.82, 83.81, 69.16, 69.15, 68.79, 61.65, 58.81, 30.56,29.34, 28.81, 28.59, 28.38, 28.29, 28.06, 28.04, 27.89, 26.05, 17.92, 17.91.

[0113] ESI + :538.71.

[0114] Example 8 Synthesis of 13-(hex-5-yn-1-yl)-10-(hex-5-yn-1-yloxy)-9-methoxy-5,6-dihydro-[1,3]dioxolo[4,5-g]isoquinolino[3,2-a]isoquinolin-7-ium (Compound 8)

[0115]

[0116] The experimental steps were the same as in Example 1, except that the first raw material was 6-bromohexyne and the second raw material was 6-bromohexyne; the NMR results of the final product were as follows:

[0117] 1 H NMR (500 MHz, DMSO-d6) δ 9.85 (s, 1H), 7.47 (s, 1H), 7.37 (d, J = 8.8Hz, 1H), 7.12 (d, J = 8.8 Hz, 1H), 6.99 (t, J = 1.1 Hz, 1H), 6.04 (s, 2H), 4.63(ddd, J = 7.6, 4.9, 1.2 Hz, 2H), 4.09 (t, J = 5.0 Hz, 2H), 3.16 (dddd, J= 19.3, 7.6, 4.6, 1.0 Hz, 2H), 2.83 (dt, J = 15.8, 7.3 Hz, 1H), 2.74 (dt, J = 15.6, 7.3Hz, 1H), 2.47 (qd, J = 5.7, 3.0 Hz, 4H), 2.06 (td, J = 3.0, 1.6 Hz, 2H), 1.85 –1.75 (m, 4H), 1.63 – 1.54 (m, 4H).

[0118] 13 C NMR (125 MHz, DMSO-d6) δ 149.57, 148.34, 147.55, 147.54, 142.81, 137.36, 134.68, 134.45, 132.47, 121.95, 121.89, 121.29, 118.04, 109.19, 108.80, 101.51, 83.79, 83.77, 69.20, 69.16, 69.15, 61.66, 58.81, 30.57, 28.67, 27.89, 27.61, 27.50, 25.28, 18.24, 18.09.

[0119] ESI + : 482.60.

[0120] Example 9 Synthesis of 13-(hex-5-yn-1-yl)-10-(hex-5-yn-1-yloxy)-9-methoxy-5,6-dihydro-[1,3]dioxolo[4,5-g]isoquinoline[3,2-a]isoquinoline-7-ium (Compound 9)

[0121]

[0122] The experimental procedure was the same as Example 1, except that the first starting material was p-methylbromobenzene and the second starting material was p-methylbromobenzene; the final product NMR results were as follows:

[0123] 1 H NMR (500 MHz, DMSO-d6) δ 9.85 (s, 1H), 7.49 (s, 1H), 7.35 (d, J = 8.5Hz, 1H), 7.30 (dt, J = 7.9, 1.1 Hz, 2H), 7.20 (d,J = 8.6 Hz, 1H), 7.18 – 7.11(m, 4H), 7.11 – 7.06 (m, 2H), 6.99 (t, J = 1.0 Hz, 1H), 6.04 (s, 2H), 5.12 (t, J = 1.0 Hz, 2H), 4.64 (ddd, J = 7.3, 4.6, 1.7 Hz, 2H), 4.19 (t, J = 1.0 Hz, 2H),3.16 (dddd, J = 19.3, 7.6, 4.6, 1.0 Hz, 2H), 2.33 (q, J = 0.9 Hz, 5H).

[0124] 13 C NMR (125 MHz, DMSO-d6) δ 149.57, 148.14, 147.54, 147.50, 142.79,138.34, 138.17, 137.13, 136.95, 133.99, 131.23, 130.86, 129.27, 129.09,128.45, 128.18, 128.03, 122.54, 121.99, 120.72, 118.17, 108.80, 108.18,101.51, 71.32, 61.66, 57.39, 36.00, 27.89, 21.05.

[0125] ESI + : 530.64.

[0126] Example 10 Synthesis of 9-methoxy-13-(4-nitrobenzyl)-10-((4-nitrobenzyl)oxy)-5,6- dihydro-[l,3]dioxolo[4,5-g]isoquinoline[3,2-a]isoquinoline-7-carboxylic acid (Compound 10)

[0127]

[0128] The experimental procedure was the same as Example 1, except that the first starting material was p-nitrobenzyl bromide and the second starting material was p-nitrobenzyl bromide; the final product NMR results were as follows:

[0129] 1H NMR (500 MHz, DMSO-d6) δ 9.85 (s, 1H), 8.21 - 8.15 (m, 3H), 8.14 - 8.08 (m, 3H), 7.61 (dt, J = 8.2, 1.0 Hz, 3H), 7.53 (t, J = 1.0 Hz, 1H), 7.54 - 7.47 (m, 3H), 7.35 (d, J = 8.5 Hz, 1H), 7.20 (d, J = 8.5 Hz, 1H), 6.99 (t, J = 1.0Hz, 1H), 6.04 (s, 3H), 5.13 (t, J = 1.1 Hz, 3H), 4.64 (ddd, J = 7.3, 4.6, 1.7 Hz,3H), 4.20 (t, J = 1.1 Hz, 3H), 3.16 (dddd, J = 19.3, 7.6, 4.6, 1.0 Hz, 3H).

[0130] 13 C NMR (125 MHz, DMSO-d6) δ 149.57, 149.54, 148.25, 148.14, 147.54, 147.50, 143.96, 142.79, 141.03, 137.13, 131.23, 130.92, 128.79, 128.68, 128.45, 123.81, 123.65, 122.54, 121.99, 120.72, 118.17, 108.80, 108.18, 101.51, 71.41, 61.65, 57.39, 36.04, 27.89.

[0131] ESI + : 592.58.

[0132] Synthesis of 13-(3,5-dimethoxybenzyl)-10-((3,5-dimethoxybenzyl)oxy)-9-methoxy-5,6-dihydro-[1,3]dioxolo[4,5-g]isoquinoline[3,2-a]isoquinoline-7-carboxylic acid (Compound 11)

[0133]

[0134] The experimental procedure was the same as in Example 1, except that the first starting material was 3,5-dimethoxybenzyl bromide and the second starting material was 3,5-dimethoxybenzyl bromide; the final product NMR results were as follows:

[0135] 1 H NMR (500 MHz, DMSO-d6) δ 9.85 (s, 1H), 7.49 (s, 1H), 7.35 (d, J = 8.5Hz, 1H), 7.20 (d, J = 8.5 Hz, 1H), 7.01 – 6.97 (m, 1H), 6.63 (dt, J = 2.3, 1.2Hz, 2H), 6.46 (dt, J = 2.1, 1.0 Hz, 2H), 6.38 (dt, J = 12.5, 2.4 Hz, 2H), 6.04(s, 2H), 5.13 (t, J = 1.0 Hz, 2H), 4.64 (ddd, J = 7.3, 4.6, 1.7 Hz, 2H), 4.08(dt, J = 9.5, 1.0 Hz, 1H), 4.01 (s, 3H), 4.03 – 3.97 (m, 1H), 3.16 (dddd, J =19.3, 7.6, 4.6, 1.0 Hz, 2H).

[0136] 13 C NMR (125 MHz, DMSO-d6) δ 160.81, 160.51, 149.57, 148.29, 147.54,147.50, 142.79, 139.48, 138.25, 137.07, 131.24, 130.70, 128.45, 122.55,121.99, 120.72, 118.17, 108.80, 108.24, 108.18, 107.23, 101.51, 99.53, 98.82,71.52, 61.65, 57.39, 55.34, 55.33, 36.36, 27.89.

[0137] ESI + :622.69.

[0138] Example 12 Synthesis of 13-allyl-10-(hept-6-yn-1 -yloxy)-9-methoxy-5,6-dihydro- [1,3]dioxolo[4,5-g]isoquinoline[3,2-a]isoquinoline-7-carboxylic acid (Compound 12)

[0139]

[0140] The experimental procedure was the same as Example 1, except that the first starting material was 7-bromoheptyne and the second starting material was 3-bromopropene; the final product NMR results were as follows:

[0141] 1 H NMR (500 MHz, DMSO-d6) δ 9.85 (s, 1H), 7.47 (s, 1H), 7.39 (d, J = 8.8Hz, 1H), 7.12 (d, J = 8.8 Hz, 1H), 6.99 (t, J = 1.0 Hz, 1H), 6.04 (s, 2H), 5.89(tt, J = 16.4, 8.1 Hz, 1H), 5.17 (ddt, J = 16.4, 2.1, 1.0 Hz, 1H), 5.07 (ddt, J =16.3, 2.1, 1.0 Hz, 1H), 4.64 (ddd, J = 7.3, 4.6, 1.7 Hz, 2H), 4.00 (t, J = 6.1Hz, 2H), 3.40 (dt, J = 8.0, 1.0 Hz, 2H), 3.16 (dddd, J = 19.3, 7.6, 4.6, 1.0 Hz,2H), 2.11 (td, J = 5.8, 3.0 Hz, 2H), 2.06 (t, J = 2.9 Hz, 1H), 1.74 (p, J = 6.3 Hz,2H), 1.57 – 1.46 (m, 4H).

[0142] 13C NMR (125 MHz, DMSO-d6) δ 149.57, 148.40, 147.57, 147.54, 142.88, 137.27, 136.32, 130.33, 130.25, 128.69, 122.35, 122.09, 121.73, 117.97, 116.29, 108.80, 108.41, 101.51, 83.79, 69.15, 68.80, 61.66, 58.15, 34.24, 29.15, 27.89, 27.85, 25.70, 17.92.

[0143] ESI + :456.56.

[0144] Example 13 Synthesis of 13-allyl-9-methoxy-10-((4-methylbenzyl)oxy)-5,6-dihydro- [l,3]dioxane[4,5-g]isoquinoline[3,2-a]isoquinoline-7-ium (Compound 13)

[0145]

[0146] The experimental procedure was the same as Example 1, except that the first starting material was p-methylbenzyl bromide and the second starting material was 3-bromopropene; the final product NMR results were as follows:

[0147] 1 H NMR (500 MHz, DMSO-d6) δ 9.85 (s, 1H), 7.47 (s, 1H), 7.39 (d, J = 0.7 Hz, 1H), 7.37 (s, 1H), 7.30 (dt, J = 7.8, 1.1 Hz, 2H), 7.20 (d, J = 8.5 Hz, 1H), 7.16 (d, J = 7.7 Hz, 2H), 6.99 (t, J = 1.1 Hz, 1H), 6.04 (s, 2H), 5.89 (tt, J = 16.5, 8.1 Hz, 1H), 5.17 (ddt, J = 16.4, 2.1, 1.0 Hz, 1H), 5.12 (t, J = 1.0 Hz, 2H), 5.07 (ddt, J= 16.3, 2.1, 1.0 Hz, 1H), 4.64 (ddd, J = 7.3, 4.6, 1.7 Hz, 2H),3.40 (dt, J = 8.0, 1.0 Hz, 2H), 3.16 (dddd, J = 19.3, 7.6, 4.6, 1.0 Hz, 2H), 2.34(d, J = 1.0 Hz, 2H).

[0148] 13 C NMR (125 MHz, DMSO-d6) δ 149.57, 148.07, 147.54, 147.47, 142.88,138.34, 137.27, 136.32, 133.99, 130.33, 130.25, 129.09, 128.69, 128.03,122.35, 121.95, 121.63, 118.20, 116.29, 108.80, 108.41, 101.51, 71.32, 61.65,58.15, 34.24, 27.89, 21.05.

[0149] ESI + : 466.56.

[0150] Example 14 Synthesis of (E)-10-(3-aminopropoxy)-13-(but-2-en-1-yl)-9-methoxy-5,6- dihydro-[1,3]dioxolo[4,5-g]isoquinoline[3,2-a]isoquinoline-7-ium (Compound 14)

[0151]

[0152] The experimental procedure was the same as Example 1, except that the first starting material was 3-bromo-1-propanamine and the second starting material was 2-buten-1-ol; the final product had the following NMR results:

[0153] 1 H NMR (500 MHz, DMSO-d6) δ 9.85 (s, 1H), 7.47 (s, 1H), 7.39 (d, J = 8.8 Hz, 1H), 6.99 (t, J = 8.8 Hz, 1H), 6.99 (t, J= 1.0 Hz, 1H), 6.04 (s, 2H), 5.87 –5.78 (m, 1H), 5.72 – 5.62 (m, 1H), 4.64 (ddd, J = 7.3, 4.6, 1.7 Hz, 2H), 4.09 –4.00 (m, 5H), 3.43 (dq, J = 7.1, 1.0 Hz, 2H), 3.16 (dddd, J = 19.3, 7.6, 4.6, 1.0Hz, 2H), 2.93 (pd, J = 5.8, 0.7 Hz, 2H), 2.81 (t, J = 6.2 Hz, 2H), 2.02 (p, J = 5.6Hz, 2H), 1.62 (dq, J = 5.3, 1.1 Hz, 3H).

[0154] 13 C NMR (125 MHz, DMSO-d6) δ 149.57, 148.39, 147.57, 147.54, 142.86,136.66, 130.43, 130.25, 128.62, 128.26, 127.61, 122.33, 122.09, 121.71,117.97, 108.80, 108.41, 101.51, 67.72, 61.65, 58.15, 37.13, 33.10, 31.06,27.89, 17.90.

[0155] ESI + :433.53.

[0156] Example 15 Synthesis of (E)-13-(but-2-en-1-yl)-9-methoxy-10-((4-nitrobenzyl)oxy)-5,6- dihydro-[1,3]dioxolo[4,5-g]isoquinoline[3,2-a]isoquinoline-7-carboxylic acid (Compound 15)

[0157]

[0158] The experimental procedure was the same as Example 1, except that the first starting material was p-nitrobenzyl bromide and the second starting material was geranyl bromide; the final product NMR results were as follows:

[0159] 1H NMR (500 MHz, DMSO-d6) δ 9.85 (s, 1H), 8.14 - 8.08 (m, 2H), 7.61 (dt, J = 8.1, 1.0 Hz, 2H), 7.47 (s, 1H), 7.41 - 7.36 (m, 1H), 7.20 (d, J = 8.5Hz, 1H), 7.01 - 6.97 (m, 1H), 6.04 (s, 2H), 5.87 - 5.78 (m, 1H), 5.72 - 5.62 (m, 1H), 5.13 (t, J = 1.1 Hz, 2H), 4.64 (ddd, J = 7.3, 4.6, 1.7 Hz, 2H), 4.01 (s, 3H), 3.43 (dq, J = 7.1, 1.0 Hz, 2H), 3.16 (dddd, J = 19.3, 7.6, 4.6, 1.0 Hz, 2H), 1.64 - 1.59 (m, 3H).

[0160] 13 C NMR (125 MHz, DMSO-d6) δ 149.57, 148.25, 148.07, 147.54, 147.47, 142.86, 141.03, 136.66, 130.42, 130.25, 128.68, 128.62, 128.26, 127.61, 123.65, 122.33, 121.95, 121.61, 118.20, 108.80, 108.41, 101.51, 71.41, 61.65, 58.15, 33.10, 27.89, 17.90.

[0161] ESI + : 511.55.

[0162] Synthesis of 10-(cyanomethoxy)-9-methoxy-13-(3-methylbut-2-en-1-yl)-5,6-dihydro- [1,3]dioxolo[4,5-g]isoquinoline[3,2-a]isoquinoline-7-ol (Compound 16)

[0163]

[0164] The experimental procedure was the same as in Example 1, except that the first starting material was bromoacetonitrile and the second starting material was 3,3-dimethylallyl bromide; the final product NMR results were as follows:

[0165] 1 H NMR (500 MHz, DMSO-d6) δ 9.84 (s, 1H), 7.47 (s, 1H), 7.39 (d, J = 8.5Hz, 1H), 7.22 (d, J = 8.8 Hz, 1H), 6.99 (t, J = 1.1 Hz, 1H), 6.04 (s, 2H), 5.31(dddd, J = 9.1, 7.3, 3.3, 1.6 Hz, 1H), 4.96 (s, 2H), 4.64 (ddd, J = 7.3, 4.6, 1.7Hz, 2H), 3.46 (dp, J = 7.4, 1.0 Hz, 2H), 3.16 (dddd, J = 19.3, 7.6, 4.6, 1.0 Hz,2H), 1.68 (q, J = 1.2 Hz, 6H).

[0166] 13 C NMR (125 MHz, DMSO-d6) δ 149.57, 147.56, 147.54, 146.69, 142.84,136.72, 130.27, 130.25, 129.75, 128.53, 123.77, 122.33, 122.00, 121.63,118.23, 116.24, 108.80, 108.41, 101.51, 61.65, 58.15, 54.47, 28.68, 27.89,24.49, 19.28.

[0167] ESI + :529.50.

[0168] Example 17 Synthesis of 9-methoxy-13-(3-methylbut-2-en-1-yl)-10-(pent-4-en-1-yloxy)- 5,6-dihydro-[1,3]dioxepino[4,5-g]isoquinoline[3,2-a]isoquinoline-7-ium (Compound 17)

[0169]

[0170] The experimental procedure was the same as in Example 1, except that the first starting material was 5-bromopentene and the second starting material was 3,3-dimethylallyl bromide; the end product NMR results are as follows:

[0171] 1 H NMR (500 MHz, DMSO-d6) δ 9.85 (s, 1H), 7.47 (s, 1H), 7.39 (d, J = 8.8Hz, 1H), 7.12 (d, J = 8.8 Hz, 1H), 6.99 (t, J = 1.1 Hz, 1H), 6.04 (s, 2H), 5.74(tt, J = 17.1, 6.8 Hz, 1H), 5.31 (dddd, J = 9.1, 7.3, 3.3, 1.6 Hz, 1H), 5.11(ddt, J = 17.1, 2.1, 1.0 Hz, 1H), 4.97 (ddt, J = 17.1, 2.1, 1.0 Hz, 1H), 4.64(ddd, J = 7.3, 4.6, 1.7 Hz, 2H), 4.09 – 4.00 (m, 4H), 3.49 – 3.43 (m, 2H), 3.16(dddd, J = 19.3, 7.6, 4.6, 1.0 Hz, 2H), 2.14 (tdt, J = 8.1, 6.9, 1.1 Hz, 2H),1.77 (tt, J = 8.4, 6.8 Hz, 2H), 1.68 (q, J = 1.2 Hz, 6H).

[0172] 13C NMR (125 MHz, DMSO-d6) δ 149.57, 148.39, 147.57, 147.54, 142.84, 137.60, 136.72, 130.25, 130.20, 129.76, 128.51, 123.77, 122.33, 122.09, 121.66, 117.97, 115.13, 108.80, 108.41, 101.51, 69.02, 61.66, 58.15, 30.17, 28.68, 28.67, 27.89, 24.49, 19.28.

[0173] ESI + : 458.58.

[0174] Synthesis of 10-(allyloxy)-13-(3-aminopropyl)-9-methoxy-5,6-dihydro-[l,3]dioxolo[4,5- g]isoquinoline[3,2-a]isoquinoline-7-ium (Compound 18)

[0175]

[0176] The experimental procedure was the same as Example 1, except that the first starting material was 3-bromopropene and the second starting material was 3-bromopropylamine; the final product NMR results are as follows:

[0177] 1 H NMR (500 MHz, DMSO-d6) δ 9.85 (s, 1H), 7.47 (s, 1H), 7.31 (d, J = 8.8 Hz, 1H), 7.17 (d, J = 8.8 Hz, 1H), 6.99 (t, J = 1.0 Hz, 1H), 6.11 - 5.99 (m, 1H), 6.04 (s, 2H), 5.39 (ddt, J = 16.8, 2.1, 1.0 Hz, 1H), 5.31 (ddt, J = 16.8, 2.2, 1.0 Hz, 1H), 4.67 - 4.58 (m, 4H), 3.16 (dddd, J = 19.3, 7.6, 4.6, 1.0 Hz, 2H), 2.87 - 2.77 (m, 4H), 1.87 (tt, J = 8.1, 5.3 Hz, 2H), 1.74 (t,J = 6.3 Hz, 2H).

[0178] 13 C NMR (125 MHz, DMSO- d 6) δ 149.57, 148.20, 147.55, 147.44, 142.81,137.12, 134.45, 132.52, 132.39, 132.27, 121.92, 121.90, 121.24, 118.50,117.80, 109.19, 108.80, 101.51, 69.80, 61.65, 58.81, 40.83, 29.70, 29.09,27.89.

[0179] ESI + :419.50.

[0180] Synthesis of 13-(3-aminopropyl)-10-((3,5-dimethoxybenzyl)oxy)-9-methoxy-5,6- dihydro-[l,3]dioxolo[4,5-g]isoquinoline[3,2-a]isoquinoline-7-carboxylic acid (Compound 19)

[0181]

[0182] The experimental procedure was the same as Example 1, except that the first starting material was 3,5-dimethoxybenzyl bromide and the second starting material was 3-bromopropylamine; the final product NMR results are as follows:

[0183] 1 H NMR (500 MHz, DMSO-d6) δ 9.85 (s, 1H), 7.47 (s, 1H), 7.37 (d, J = 8.8Hz, 1H), 7.20 (d, J = 8.5 Hz, 1H), 6.99 (t, J = 1.0 Hz, 1H), 6.63 (dt, J = 2.3, 1.2Hz, 2H), 6.39 (t, J = 2.4 Hz, 1H), 6.04 (s, 2H), 5.13 (t, J = 1.0 Hz, 2H), 4.63(ddd, J = 7.6, 4.9, 1.2 Hz, 2H), 4.01 (s, 3H), 3.16 (dddd, J= 19.3, 7.6, 4.6,1.0 Hz, 2H), 2.87 – 2.77 (m, 4H), 1.87 (tt, J = 8.1, 5.3 Hz, 2H), 1.74 (t, J =6.3 Hz, 2H).

[0184] 13 C NMR (125 MHz, DMSO-d6) δ 160.51, 149.57, 148.15, 147.55, 147.37, 142.81, 138.25, 137.12, 134.45, 132.52, 132.39, 121.92, 121.81, 121.33, 118.17, 109.19, 108.80, 107.23, 101.51, 99.53, 71.52, 61.65, 58.81, 55.33, 40.83, 29.70, 29.09, 27.89.

[0185] ESI + : 529.61.

[0186] Example 20 Synthesis of 9-methoxy-10-((3-methylbut-2-en-1-yl)oxy)-13-pentyl-5,6-dihydro-[1,3]dioxolo[4,5-g]isoquinoline[3,2-a]isoquinoline-7-carboxylic acid (Compound 20)

[0187]

[0188] The experimental procedure was the same as Example 1, except that the first starting material was bromoacetonitrile and the second starting material was 3,3-dimethylallyl bromide; the final product NMR results were as follows:

[0189] 1 H NMR (500 MHz, DMSO-d6) δ 9.85 (s, 1H), 7.47 (s, 1H), 7.31 (d, J = 8.8Hz, 1H), 7.17 (d, J = 8.8 Hz, 1H), 6.99 (t, J = 1.0 Hz, 1H), 6.04 (s, 2H), 5.28(tp, J = 4.8, 1.6 Hz, 1H), 4.67 – 4.59 (m, 4H), 3.16 (dddd, J= 19.3, 7.6, 4.6,1.0 Hz, 2H), 2.86 – 2.70 (m, 2H), 1.75 (q, J = 1.1 Hz, 6H), 1.73 – 1.63 (m,2H), 1.35 (dddd, J = 8.8, 7.4, 6.2, 3.5 Hz, 4H), 0.92 – 0.86 (m, 3H).

[0190] 13 C NMR (125 MHz, DMSO-d6) δ 149.57, 148.27, 147.55, 147.44, 142.81,137.69, 137.36, 134.69, 134.45, 132.47, 121.89, 121.21, 119.15, 117.80,109.19, 108.80, 101.51, 66.11, 61.65, 58.81, 32.42, 30.44, 28.24, 27.89,24.62, 23.18, 19.90, 14.04.

[0191] ESI + :460.59.

[0192] Example 21 Synthesis of 9-methoxy-10-((3-methylbut-2-en-1-yl)oxy)-13-pentyl-5,6-dihydro- [1,3]dioxolo[4,5-g]isoquinoline[3,2-a]isoquinoline-7-carboxylic acid (Compound 21)

[0193]

[0194] The experimental procedure was the same as Example 1, except that the first starting material was 6-bromohexyne and the second starting material was 5-bromo-n-pentane; the final product NMR results were as follows:

[0195] 1 H NMR (500 MHz, DMSO-d6) δ 9.85 (s, 1H), 7.47 (s, 1H), 7.37 (d, J = 8.8Hz, 1H), 7.12 (d, J = 8.8 Hz, 1H), 6.99 (t, J = 1.0 Hz, 1H), 6.04 (s, 2H), 4.63(ddd, J= 7.6, 4.9, 1.2 Hz, 2H), 4.09 (t, J = 5.0 Hz, 2H), 3.16 (dddd, J = 19.3,7.6, 4.6, 1.0 Hz, 2H), 2.86 – 2.70 (m, 2H), 2.47 (td, J = 5.9, 3.0 Hz, 2H),2.06 (s, 1H), 2.06 (d, J = 6.1 Hz, 0H), 1.81 (tt, J = 7.5, 5.0 Hz, 2H), 1.68 (tt, J = 8.2, 7.3 Hz, 2H), 1.58 (tt, J = 7.5, 5.9 Hz, 2H), 1.35 (dddd, J = 8.8, 7.4,6.1, 3.5 Hz, 4H), 0.92 – 0.86 (m, 3H).

[0196] 13 C NMR (125 MHz, DMSO-d6) δ 149.57, 148.34, 147.55, 147.54, 142.81,137.36, 134.69, 134.45, 132.47, 121.95, 121.89, 121.29, 118.04, 109.19,108.80, 101.51, 83.76, 69.20, 69.15, 61.65, 58.81, 32.42, 30.44, 28.67,28.24, 27.89, 25.28, 23.18, 18.09, 14.04.

[0197] ESI + : 472.60.

[0198] Synthesis of 13-(hept-6-en-1-yl)-9-methoxy-10-(pentyloxy)-5,6-dihydro-[1,3]dioxolo[4,5-g]isoquinoline[3,2-a]isoquinoline-7-carboxylic acid (Compound 22)

[0199]

[0200] The experimental procedure was the same as Example 1, except that the first starting material was 5-bromo-n-pentane and the second starting material was 7-bromo-n-heptene; the final product NMR results were as follows:

[0201] 1 H NMR (500 MHz, DMSO-d6) δ 9.85 (s, 1H), 7.47 (s, 1H), 7.37 (d, J = 8.8Hz, 1H), 7.12 (d, J = 8.8 Hz, 1H), 6.99 (t, J = 1.0 Hz, 1H), 6.04 (s, 2H), 5.77(tt, J = 17.1, 6.8 Hz, 1H), 5.12 (ddt, J = 17.1, 2.1, 1.0 Hz, 1H), 4.97 (ddt, J =17.1, 2.1, 1.1 Hz, 1H), 4.63 (ddd, J = 7.6, 4.9, 1.2 Hz, 2H), 4.02 (d, J = 11.5Hz, 1H), 3.16 (dddd, J = 19.3, 7.6, 4.6, 1.0 Hz, 2H), 2.78 (t, J = 8.5 Hz, 2H),2.03 (tdt, J = 7.8, 6.7, 1.1 Hz, 2H), 1.78 (tt, J = 7.3, 5.6 Hz, 2H), 1.69 (tt, J =8.5, 6.8 Hz, 2H), 1.46 – 1.37 (m, 2H), 1.41 – 1.30 (m, 5H), 1.34 (s, 2H),0.90 (t, J = 6.8 Hz, 3H).

[0202] 13C NMR (125 MHz, DMSO-d6) δ 149.57, 148.34, 147.55, 147.54, 142.81, 139.08, 137.36, 135.30, 134.45, 132.47, 121.95, 121.89, 121.29, 118.04, 114.41, 109.19, 108.80, 101.51, 69.01, 61.65, 58.81, 33.79, 30.56, 29.05, 28.72, 28.49, 28.44, 28.26, 27.89, 22.50, 13.97.

[0203] ESI + : 488.65.

[0204] Synthesis of 13-(hept-6-en-1-yl)-10-(hex-5-yn-1-yloxy)-9-methoxy-5,6-dihydro- [1,3]dioxolo[4,5-g]isoquinoline[3,2-a]isoquinoline-7-ol (Compound 23)

[0205]

[0206] The experimental procedure was the same as Example 1, except that the first starting material was 6-bromo-hex-1-yne and the second starting material was 7-bromo- hept-1-ene; the final product NMR results are as follows:

[0207] 1 H NMR (500 MHz, DMSO-d6) δ 9.85 (s, 1H), 7.47 (s, 1H), 7.37 (d, J = 8.8Hz, 1H), 7.12 (d, J = 8.8 Hz, 1H), 6.99 (t, J = 1.0 Hz, 1H), 6.04 (s, 2H), 5.77(tt, J = 17.1, 6.8 Hz, 1H), 5.12 (ddt, J = 17.1, 2.1, 1.0 Hz, 1H), 4.97 (ddt, J =17.1, 2.1, 1.1 Hz, 1H), 4.63 (ddd, J = 7.6, 4.9, 1.2 Hz, 2H), 4.09 (t, J = 5.0Hz, 2H), 3.16 (dddd,J = 19.3, 7.6, 4.6, 1.0 Hz, 2H), 2.78 (t, J = 8.5 Hz, 2H),2.47 (td, J = 5.9, 3.0 Hz, 2H), 2.09 – 1.99 (m, 3H), 1.81 (tt, J = 7.5, 5.0 Hz,2H), 1.69 (tt, J = 8.4, 6.8 Hz, 2H), 1.58 (tt, J = 7.5, 5.9 Hz, 2H), 1.39 – 1.30(m, 2H), 1.34 (s, 2H).

[0208] 13 C NMR (125 MHz, DMSO-d6) δ 149.57, 148.34, 147.55, 147.54, 142.81,139.08, 137.36, 135.30, 134.45, 132.47, 121.95, 121.89, 121.29, 118.04,114.41, 109.19, 108.80, 101.51, 83.76, 69.20, 69.15, 61.66, 58.81, 33.79,30.56, 28.72, 28.67, 28.49, 28.44, 27.89, 25.28, 18.09.

[0209] ESI + :498.64.

[0210] Example 24 Synthesis of 9-methoxy-13-(pent-4-en-1-yl)-10-(pentyloxy)-5,6-dihydro-[1,3]dioxolo[4,5-g]isoquinolino[3,2-a]isoquinolin-7-ium (Compound 24)

[0211]

[0212] The experimental steps were the same as in Example 1, except that the first raw material was 5-bromopentane and the second raw material was 5-bromopentene. The NMR results of the final product were as follows:

[0213] 1 H NMR (500 MHz, DMSO-d6) δ 9.85 (s, 1H), 7.47 (s, 1H), 7.37 (d, J= 8.8 Hz, 1H), 6.99 (t, J = 8.8 Hz, 1H), 6.99 (t, J = 1.0 Hz, 1H), 6.04 (s, 2H), 5.71(tt, J = 17.2, 6.8 Hz, 1H), 5.12 (ddt, J = 17.1, 2.1, 1.0 Hz, 1H), 4.97 (ddt, J =17.1, 2.1, 1.0 Hz, 1H), 4.63 (ddd, J = 7.6, 4.9, 1.2 Hz, 2H), 4.02 (d, J = 11.5Hz, 1H), 4.02 (s, 4H), 3.16 (dddd, J = 19.3, 7.6, 4.6, 1.0 Hz, 2H), 2.88 (s,1H), 2.85 (s, 1H), 2.14 (tdt, J = 8.0, 7.0, 1.1 Hz, 2H), 1.78 (tt, J = 7.3, 5.6Hz, 2H), 1.65 (tt, J = 9.3, 7.9 Hz, 2H), 1.46 – 1.31 (m, 5H), 0.90 (t, J = 6.8Hz, 3H).

[0214] 13 C NMR (125 MHz, DMSO-d6) δ 149.57, 148.34, 147.55, 147.54, 142.81,138.25, 137.10, 134.45, 133.11, 132.48, 121.95, 121.91, 121.29, 118.04,114.91, 109.19, 108.80, 101.51, 69.01, 61.65, 58.81, 33.02, 30.13, 29.05,28.26, 27.89, 27.85, 22.50, 13.97.

[0215] ESI + :460.59.

[0216] Example 25 Synthesis of 9-methoxy-10-(oct-7-yn-1-yloxy)-13-(pent-4-en-1-yl)-5,6- dihydro-[1,3]dioxolo[4,5-g]isoquinoline[3,2-a]isoquinoline-7-carboxylic acid (Compound 25)

[0217]

[0218] The experimental procedure was the same as Example 1, except that the first starting material was 8-bromo-oct-7-yne and the second starting material was 5-bromo-pent-1 - ene; the final product NMR results were as follows:

[0219] 1 H NMR (500 MHz, DMSO-d6) δ 9.85 (s, 0H), 7.47 (s, 0H), 7.37 (d, J = 8.8Hz, 0H), 7.12 (d, J = 8.8 Hz, 0H), 6.99 (t, J = 1.0 Hz, 0H), 6.04 (s, 1H), 5.71(tt, J = 17.2, 6.8 Hz, 0H), 5.12 (ddt, J = 17.1, 2.1, 1.0 Hz, 0H), 4.97 (ddt, J =17.1, 2.1, 1.0 Hz, 0H), 4.63 (ddd, J = 7.6, 4.9, 1.2 Hz, 1H), 4.04 – 3.97 (m,2H), 3.16 (dddd, J = 19.3, 7.6, 4.6, 1.0 Hz, 1H), 2.18 – 2.04 (m, 3H), 1.77(tt, J = 7.5, 6.1 Hz, 1H), 1.65 (tt, J = 9.3, 7.9 Hz, 1H), 1.55 – 1.34 (m, 3H).

[0220] 13C NMR (125 MHz, DMSO-d6) δ 149.57, 148.34, 147.55, 147.54, 142.81, 138.25, 137.10, 134.45, 133.11, 132.48, 121.95, 121.91, 121.29, 118.04, 114.91, 109.19, 108.80, 101.51, 83.81, 69.15, 68.79, 61.66, 58.81, 33.02, 30.13, 29.34, 28.28, 28.06, 27.89, 27.85, 26.05, 17.91.

[0221] ESI + :498.64.

[0222] Synthesis of 10-(allyloxy)-9-methoxy-13-(oct-7-yn-1-yl)-5,6-dihydro-[1,3] dioxolo[4,5-g]isoquinoline[3,2-a]isoquinoline-7-ol (Compound 26)

[0223]

[0224] The experimental procedure was same as Example 1, except that the first starting material was 3-bromopropene and the second starting material was 8-bromo-n-octyne; the final product NMR results were as follows:

[0225] 1 H NMR (500 MHz, DMSO-d6) δ 9.85 (s, 1H), 7.47 (s, 1H), 7.31 (d, J = 8.8Hz, 1H), 7.17 (d, J = 8.8 Hz, 1H), 7.01 – 6.97 (m, 1H), 6.11 – 5.99 (m, 1H),6.04 (s, 2H), 5.39 (ddt, J = 16.8, 2.2, 1.0 Hz, 1H), 5.31 (ddt, J = 16.9, 2.2,1.0 Hz, 1H), 4.63 (ddd, J = 7.6, 4.9, 1.2 Hz, 2H), 4.61 (dt, J = 5.4, 1.0 Hz,2H), 3.16 (dddd, J= 19.3, 7.6, 4.6, 1.0 Hz, 2H), 2.78 (t, J = 8.5 Hz, 2H), 2.12(td, J = 6.0, 3.1 Hz, 2H), 2.06 (t, J = 2.9 Hz, 1H), 1.68 (tt, J = 8.4, 7.2 Hz,2H), 1.52 – 1.44 (m, 2H), 1.44 – 1.33 (m, 4H).

[0226] 13 C NMR (125 MHz, DMSO-d6) δ 149.57, 148.20, 147.55, 147.44, 142.81, 137.36, 135.30, 134.45, 132.47, 132.27, 121.89, 121.21, 118.50, 117.80, 109.19, 108.80, 101.51, 83.81, 69.80, 69.15, 61.65, 58.81, 30.56, 28.81, 28.59, 28.38, 28.04, 27.89, 17.91.

[0227] ESI + : 470.59.

[0228] Synthesis of 9-methoxy-10-((4-methylbenzyl)oxy)-13-(oct-7-yn-1-yl)-5,6-dihydro- [1,3]dioxolo[4,5-g]isoquinoline[3,2-a]isoquinoline-7-carboxylic acid (Compound 27)

[0229]

[0230] The experimental procedure was same as Example 1, except that the first starting material was p-methylbenzyl bromide and the second starting material was 8-bromo-n-octyn; the final product NMR results were as follows:

[0231] 1 H NMR (500 MHz, DMSO-d6) δ 9.85 (s, 1H), 7.47 (s, 1H), 7.37 (d, J = 8.8Hz, 1H), 7.30 (dt, J = 7.8, 1.1 Hz, 2H), 7.20 (d, J= 7.6, 4.9, 1.2 Hz, 2H), 3.16 (dddd, J = 7.6, 4.9, 1.2 Hz, 2H), 3.16 (dddd, J = 7.6, 4.9, 1.2 Hz, 2H), 3.16 (dddd, J = 7.6, 4.9, 1.2 Hz, 2H), 3.16 (dddd, J = 7.6, 4.9, 1.2 Hz, 2H), 3.16 (dddd, J = 7.6, 4.9, 1.2 Hz, 2H), 3.16 (dddd, J = 7.6, 4.9, 1.2 Hz, 2H), 3.16 (dddd, J = 7.6, 4.9, 1.2 Hz, 2H), 3.16 (dddd, J = 7.6, 4.9, 1.2 Hz, 2H), 3.16 (dddd, J = 7.6, 4.9, 1.2 Hz, 2H), 3.16 (dddd, J = 7.6, 4.9, 1.2 Hz, 2H), 3.16 (dddd,

[0232] 13 C NMR (125 MHz, DMSO-d6) δ 149.57, 148.00, 147.55, 147.37, 142.81,138.34, 137.36, 135.30, 134.45, 133.99, 132.47, 129.09, 128.03, 121.89,121.80, 121.25, 118.15, 109.19, 108.80, 101.51, 83.81, 71.32, 69.15, 61.65,58.81, 30.56, 28.81, 28.59, 28.38, 28.04, 27.89, 21.05, 17.91.

[0233] ESI + : 534.68.

[0234] Synthesis of 13-(hex-5-yn-1-yl)-9-methoxy-10-((3-methylbut-2-en-1-yl)oxy)-5,6- dihydro-[1,3]dioxolo[4,5-g]isoquinoline[3,2-a]isoquinoline-7-ol (Compound 28)

[0235]

[0236] The experimental procedure was the same as in Example 1, except that the first starting material was 3,3-dimethylallyl bromide and the second starting material was 6-bromo-n-hexynyl; the final product NMR results were as follows:

[0237] 1 H NMR (500 MHz, DMSO-d6) δ 9.85 (s, 1H), 7.47 (s, 1H), 7.31 (d, J = 8.7Hz, 1H), 7.17 (d, J = 8.8 Hz, 1H), 6.99 (t, J = 1.0 Hz, 1H), 6.04 (s, 2H), 5.28(ddp, J = 6.5, 3.2, 1.6 Hz, 1H), 4.67 – 4.59 (m, 4H), 3.16 (dddd, J = 19.3, 7.6,4.6, 1.0 Hz, 2H), 2.83 (dt, J = 15.8, 7.3 Hz, 1H), 2.74 (dt, J = 15.6, 7.3 Hz,1H), 2.46 (td, J = 5.7, 3.0 Hz, 2H), 2.06 (t, J = 3.1 Hz, 1H), 1.79 (tt, J = 7.5,6.1 Hz, 2H), 1.75 (q, J = 1.1 Hz, 6H), 1.59 (p, J = 5.8 Hz, 2H).

[0238] 13 C NMR (125 MHz, DMSO-d6) δ 149.57, 148.27, 147.55, 147.44, 142.81,137.69, 137.36, 134.70, 134.45, 132.47, 121.89, 121.21, 119.15, 117.80,109.19, 108.80, 101.51, 83.79, 69.15, 66.11, 61.65, 58.81, 30.57, 27.89,27.61, 27.50, 24.62, 19.90, 18.24.

[0239] ESI+ :470.59.

[0240] Synthesis of 10-(3-aminopropoxy)-13-(hex-5-yn-1-yl)-9-methoxy-5,6-dihydro- [1,3]dioxolo[4,5-g]isoquinoline[3,2-a]isoquinoline-7-ol (Compound 29)

[0241]

[0242] The experimental procedure was the same as Example 1, except that the first starting material was 3-bromopropylamine and the second starting material was 6-bromo-hex-1-yn; the final product NMR results were as follows:

[0243] 1 H NMR (500 MHz, DMSO-d6) δ 9.85 (s, 1H), 7.47 (s, 1H), 7.37 (d, J = 8.8Hz, 1H), 7.12 (d, J = 8.8 Hz, 1H), 6.99 (t, J = 1.0 Hz, 1H), 6.04 (s, 2H), 4.63(ddd, J = 7.6, 4.9, 1.2 Hz, 2H), 4.09 – 4.00 (m, 5H), 3.16 (dddd, J = 19.3, 7.6,4.6, 1.0 Hz, 2H), 2.97 – 2.89 (m, 2H), 2.88 – 2.78 (m, 3H), 2.74 (dt, J = 15.6,7.3 Hz, 1H), 2.46 (td, J = 5.7, 3.0 Hz, 2H), 2.06 (t, J = 3.1 Hz, 1H), 2.02 (p, J =5.6 Hz, 2H), 1.79 (tt, J = 7.4, 6.1 Hz, 2H), 1.59 (p, J = 5.9 Hz, 2H).

[0244] 13C NMR (125 MHz, DMSO-d6) δ 149.57, 148.33, 147.55, 147.54, 142.81, 137.36, 134.68, 134.45, 132.47, 121.95, 121.89, 121.29, 118.04, 109.19, 108.80, 101.51, 83.79, 69.15, 67.72, 61.65, 58.81, 37.13, 31.06, 30.57, 27.89, 27.61, 27.50, 18.24.

[0245] ESI + : 459.57.

[0246] Synthesis of 10-(allyloxy)-13-(cyanomethyl)-9-methoxy-5,6-dihydro-[l,3]dioxolo[4,5- g]isoquinoline[3,2-a]isoquinoline-7-carboxylic acid (Compound 30)

[0247]

[0248] The experimental procedure was the same as Example 1, except that the first starting material was 3-bromopropene and the second starting material was bromoacetonitrile; the final product NMR results were as follows:

[0249] 1 H NMR (500 MHz, DMSO-d6) δ 9.85 (s, 1H), 7.47 (s, 1H), 7.28 (d, J = 8.5 Hz, 1H), 7.19 (d, J = 8.8 Hz, 1H), 6.99 (t, J = 1.0 Hz, 1H), 6.11 - 5.99 (m, 1H), 6.04 (s, 2H), 5.39 (ddt, J = 16.8, 2.1, 1.0 Hz, 1H), 5.31 (ddt, J = 16.9, 2.2, 1.0 Hz, 1H), 4.66 - 4.58 (m, 4H), 4.20 (s, 2H), 3.16 (dddd, J = 17.3, 7.3, 4.6, 1.0 Hz, 2H).

[0250] 13C NMR (125 MHz, DMSO-d6) δ 149.57, 148.32, 147.62, 147.51, 142.71, 136.13, 132.27, 130.26, 128.89, 127.79, 122.75, 122.04, 121.46, 118.50, 117.91, 117.11, 108.80, 108.36, 101.51, 69.80, 61.65, 58.16, 27.89, 18.88.

[0251] ESI + : 401.44.

[0252] Synthesis of 13-(cyanomethyl)-9-methoxy-10-((4-nitrobenzyl)oxy)-5,6-dihydro- [l,3]dioxolo[4,5-g]isoquinoline[3,2-a]isoquinoline-7-carboxylic acid (Compound 31)

[0253]

[0254] The experimental procedure was the same as Example 1, except that the first starting material was p-nitrobenzyl bromide and the second starting material was bromoacetonitrile; the final product NMR results were as follows:

[0255] 1 H NMR (500 MHz, DMSO-d6) δ 9.85 (s, 1H), 8.14 - 8.08 (m, 2H), 7.61 (dt, J = 8.1, 1.0 Hz, 2H), 7.47 (s, 1H), 7.36 (d, J = 8.8 Hz, 1H), 7.18 (d, J = 8.5 Hz, 1H), 6.99 (t, J = 1.0 Hz, 1H), 6.04 (s, 2H), 5.13 (t, J = 1.1 Hz, 2H), 4.63 (ddd, J = 7.3, 4.6, 1.1 Hz, 2H), 4.20 (s, 2H), 3.16 (dddd, J = 17.3, 7.3, 4.6, 1.0 Hz, 2H).

[0256] 13C NMR (125 MHz, DMSO-d6) δ 149.57, 148.25, 148.14, 147.74, 147.51, 142.71, 141.03, 136.13, 130.26, 128.89, 128.68, 127.79, 123.65, 122.75, 121.97, 121.52, 118.17, 117.11, 108.80, 108.36, 101.51, 71.41, 61.65, 58.16, 27.89, 18.88.

[0257] ESI + : 496.50.

[0258] Example 32 Synthesis of 9-methoxy-13-(4-methylbenzyl)-10-((3-methylbut-2-en-1-yl)oxy)-5,6-dihydro-[1,3]dioxolo[4,5-g]isoquinoline[3,2-a]isoquinoline-7-carboxylic acid (Compound 32)

[0259]

[0260] The experimental procedure was the same as in Example 1, except that the first starting material was 3,3-dimethylallyl bromide and the second starting material was p-methylbenzyl bromide; the final product NMR results are as follows:

[0261] 1 H NMR (500 MHz, DMSO-d6) δ 9.85 (s, 1H), 7.49 (s, 1H), 7.28 (d, J = 8.7 Hz, 1H), 7.18 (d, J = 8.5 Hz, 1H), 7.14 (dq, J = 8.0, 1.2 Hz, 2H), 7.11 - 7.06 (m, 2H), 6.99 (t, J = 1.0 Hz, 1H), 6.04 (s, 2H), 5.28 (ddp, J = 4.8, 3.1, 1.6 Hz, 1H), 4.67 - 4.59 (m, 4H), 4.19 (t, J = 1.0 Hz, 2H), 3.16 (dddd, J = 19.3, 7.6, 4.6, 1.0 Hz, 2H), 2.33 (d, J = 1.1 Hz, 2H), 1.75 (q,J = 1.1 Hz, 6H).

[0262] 13 C NMR (125 MHz, DMSO-d6) δ 149.57, 148.39, 147.57, 147.54, 142.79,138.17, 137.69, 137.13, 136.95, 131.23, 130.86, 129.27, 128.45, 128.18,122.54, 122.05, 120.67, 119.15, 117.81, 108.80, 108.18, 101.51, 66.11, 61.65,57.39, 36.00, 27.89, 24.62, 21.04, 19.90.

[0263] ESI + : 494.61.

[0264] Synthesis of 10-(hex-5-yn-1-yloxy)-9-methoxy-13-(4-methylbenzyl)-5,6-dihydro- [1,3]dioxolo[4,5-g]isoquinoline[3,2-a]isoquinoline-7-ol (Compound 33)

[0265]

[0266] The experimental procedure was the same as Example 1, except that the first starting material was 6-bromohex-1-yne and the second starting material was p-methylbromobenzene; the final product NMR results were as follows:

[0267] 1 H NMR (500 MHz, DMSO-d6) δ 9.85 (s, 0H), 7.49 (s, 0H), 7.34 (d, J = 8.8Hz, 0H), 7.17 – 7.06 (m, 2H), 6.04 (s, 1H), 4.64 (ddd, J = 7.3, 4.6, 1.7 Hz,1H), 4.19 (t, J = 1.0 Hz, 1H), 4.09 (t, J = 5.0 Hz, 1H), 3.16 (dddd, J = 19.3, 7.6,4.6, 1.0 Hz, 1H), 2.47 (td, J = 5.9, 3.0 Hz, 1H), 1.81 (tt,J = 7.5, 5.0 Hz, 1H),1.58 (tt, J = 7.5, 5.9 Hz, 1H).

[0268] 13 C NMR (125 MHz, DMSO-d6) δ 149.57, 148.46, 147.63, 147.54, 142.79,138.17, 137.13, 136.95, 131.18, 130.86, 129.27, 128.45, 128.18, 122.54,122.09, 120.60, 117.92, 108.80, 108.18, 101.51, 83.76, 69.20, 69.15, 61.65,57.39, 36.00, 28.67, 27.89, 25.28, 21.04, 18.09.

[0269] ESI + :506.62.

[0270] Example 34 Synthesis of 9-methoxy-13-(4-nitrobenzyl)-10-(pentyloxy)-5,6-dihydro- [l,3]dioxepino[4,5-g]isoquinoline[3,2-a]isoquinoline-7-ium (Compound 34)

[0271]

[0272] The experimental procedure was the same as Example 1, except that the first starting material was 5-bromo-n-pentane and the second starting material was p-nitrobenzyl bromide; the final product NMR results were as follows:

[0273] 1 H NMR (500 MHz, DMSO-d6) δ 9.85 (s, 1H), 8.21 – 8.15 (m, 2H), 7.55 –7.47 (m, 3H), 7.34 (d, J = 8.8 Hz, 1H), 7.12 (d, J = 8.8 Hz, 1H), 6.99 (t, J = 1.0Hz, 1H), 6.04 (s, 2H), 4.64 (ddd, J = 7.3, 4.6, 1.7 Hz, 2H), 4.20 (t, J = 1.1 Hz,2H), 4.02 (d, J= 11.5 Hz, 1H), 3.16 (dddd, J = 19.3, 7.6, 4.6, 1.0 Hz, 2H), 1.78(tt, J = 7.4, 5.6 Hz, 2H), 1.46 – 1.31 (m, 4H), 0.90 (t, J = 6.8 Hz, 3H).

[0274] 13 C NMR (125 MHz, DMSO-d6) δ 149.57, 149.54, 148.46, 147.63, 147.54,143.96, 142.79, 137.13, 131.23, 130.92, 128.79, 128.45, 123.81, 122.54,122.09, 120.60, 117.92, 108.80, 108.18, 101.51, 69.01, 61.65, 57.39, 36.04,29.05, 28.26, 27.89, 22.50, 13.97.

[0275] ESI + :527.60.

[0276] Example 35 Synthesis of 9-methoxy-13-(4-nitrobenzyl)-10-(oct-7-yn-1-yloxy)-5,6-dihydro-[1,3]dioxolo[4,5-g]isoquinolino[3,2-a]isoquinolin-7-ium (Compound 35)

[0277]

[0278] The experimental steps were the same as in Example 1, except that the first raw material was 8-bromooctyne and the second raw material was p-nitrobenzyl bromide. The NMR results of the final product were as follows:

[0279] 1 H NMR (500 MHz, DMSO-d6) δ 9.85 (s, 1H), 8.21 – 8.15 (m, 2H), 7.55 –7.47 (m, 3H), 7.34 (d, J = 8.8 Hz, 1H), 7.12 (d, J = 8.8 Hz, 1H), 7.01 – 6.97 (m,1H), 6.04 (s, 2H), 4.64 (ddd, J= 7.3, 4.6, 1.7 Hz, 2H), 4.20 (t, J = 1.1 Hz,2H), 4.00 (t, J = 6.1 Hz, 2H), 3.16 (dddd, J = 19.3, 7.6, 4.6, 1.0 Hz, 2H), 2.12(td, J = 6.0, 3.0 Hz, 2H), 2.06 (t, J = 2.9 Hz, 1H), 1.77 (tt, J = 7.5, 6.1 Hz,2H), 1.55 – 1.34 (m, 6H).

[0280] 13 C NMR (125 MHz, DMSO-d6) δ 149.57, 149.54, 148.46, 147.63, 147.54,143.96, 142.79, 137.13, 131.23, 130.92, 128.79, 128.45, 123.81, 122.54,122.09, 120.60, 117.92, 108.80, 108.18, 101.51, 83.81, 69.15, 68.79, 61.65,57.39, 36.04, 29.34, 28.28, 28.06, 27.89, 26.05, 17.91.

[0281] ESI + :565.65.

[0282] Synthesis of 10-(allyloxy)-13-(3,5-dimethoxybenzyl)-9-methoxy-5,6-dihydro- [l,3]dioxolo[4,5-g]isoquinoline[3,2-a]isoquinoline-7(8H)-one (Compound 36)

[0283]

[0284] The experimental procedure was the same as in Example 1, except that the first starting material was 3-bromopropene and the second starting material was 3,5- dimethoxybenzyl bromide; the final product NMR results were as follows:

[0285] 1 H NMR (500 MHz, DMSO-d6) δ 9.85 (s, 1H), 7.49 (s, 1H), 7.28 (d, J= 8.8Hz, 1H), 7.17 (d, J = 8.5 Hz, 1H), 6.99 (t, J = 1.0 Hz, 1H), 6.46 (dt, J = 2.2, 1.0Hz, 2H), 6.36 (t, J = 2.4 Hz, 1H), 6.11 – 5.99 (m, 1H), 6.04 (s, 2H), 5.39(ddt, J = 16.8, 2.2, 1.0 Hz, 1H), 5.31 (ddt, J = 16.8, 2.2, 1.0 Hz, 1H), 4.67 –4.58 (m, 5H), 4.08 (dt, J = 9.5, 1.0 Hz, 1H), 4.03 – 3.97 (m, 1H), 3.79 (s,6H), 3.16 (dddd, J = 19.3, 7.6, 4.6, 1.0 Hz, 2H).

[0286] 13 C NMR (125 MHz, DMSO-d6) δ 160.81, 149.57, 148.32, 147.57, 147.54,142.79, 139.48, 137.07, 132.27, 131.24, 130.70, 128.45, 122.55, 122.05,120.68, 118.50, 117.81, 108.80, 108.24, 108.18, 101.51, 98.82, 69.80, 61.65,57.39, 55.33, 36.36, 27.89.

[0287] ESI + :512.58.

[0288] Example 37 Synthesis of 13-(3,5-dimethoxybenzyl)-9-methoxy-10-(prop-2-yn-1-yloxy)-5,6-dihydro-[1,3]dioxolo[4,5-g]isoquinolino[3,2-a]isoquinolin-7-ium (Compound 37)

[0289]

[0290] The experimental procedure was the same as in Example 1, except that the first starting material was 3-bromopropyne and the second starting material was 3,5-dimethoxybenzyl bromide; the final product had the following NMR results:

[0291] 1 H NMR (500 MHz, DMSO-d6) δ 9.84 (s, 1H), 7.49 (s, 1H), 7.34 (d, J = 8.8Hz, 1H), 7.23 (d, J = 8.8 Hz, 1H), 6.99 (t, J = 1.0 Hz, 1H), 6.46 (dt, J = 2.2, 1.0Hz, 2H), 6.36 (t, J = 2.4 Hz, 1H), 6.04 (s, 2H), 4.83 (d, J = 3.2 Hz, 2H), 4.64(ddd, J = 7.3, 4.6, 1.7 Hz, 2H), 4.08 (dt, J = 9.5, 1.0 Hz, 1H), 4.00 (s, 3H),4.00 (dt, J = 9.3, 1.0 Hz, 1H), 3.79 (s, 6H), 3.36 (d, J = 6.1 Hz, 1H), 3.16(dddd, J = 19.3, 7.6, 4.6, 1.0 Hz, 2H).

[0292] 13 C NMR (125 MHz, DMSO-d6) δ 160.81, 149.57, 147.54, 147.52, 146.74,142.79, 139.48, 137.07, 131.25, 130.70, 128.45, 122.55, 122.00, 120.80,118.52, 108.80, 108.24, 108.18, 101.51, 98.82, 78.41, 76.70, 61.65, 58.28,57.39, 55.33, 36.36, 27.89.

[0293] ESI + :510.57.

[0294] Example 38 Anti-Candida albicans activity of 10,13-disubstituted berberine derivatives

[0295] 1) Preparation of bacteria solution

[0296] Candida albicans SC5314 was prepared into a bacteria solution of 2 x 10 4 CFU / ml with RPMI1640 medium by counting with a hemocytometer. 4

[0297] 2) Preparation of 10,13-disubstituted berberine derivative solution

[0298] The 10,13-disubstituted berberine derivative was prepared into a stock solution of 10 mg / mL with sterile DMSO as solvent, and then diluted with sterile DMSO to obtain dilutions of 128 μg / mL, 64 μg / mL, 32 μg / mL, 16 μg / mL, 8 μg / mL, 4 μg / mL, 2 μg / mL, 1 μg / mL, 0.5 μg / mL, 0.25 μg / mL, and 0.125 μg / mL.

[0299] The positive control drugs (fluconazole, amphotericin B) were prepared into a stock solution of 100 μg / mL with sterile DMSO as solvent, and then diluted with sterile DMSO to obtain dilutions of 4 μg / mL, 2 μg / mL, 1 μg / mL, 0.5 μg / mL, 0.25 μg / mL, 0.125 μg / mL, 0.0625 μg / mL, and 0.03125 μg / mL as positive control drugs for detection of Candida albicans.

[0300] 3) Determination of minimum inhibitory concentration of 10,13-disubstituted berberine derivatives against Candida albicans

[0301] a. Take a 96-well cell culture plate, and add 100 μL RPMI1640 medium and 100 μL prepared bacteria solution to each well of the control group;

[0302] b. Add 100 μL prepared bacteria solution and 100 μL positive control drug dilution of different dilution gradients to each well of the positive control group;

[0303] Add 100 μL prepared bacteria solution and 100 μL compound dilution of different dilution gradients to each well of the experimental group;

[0304] Add an equal amount of sterile DMSO to each well of the negative control group;

[0305] ​c. After 24 hours of incubation at 37°C, observe the growth of C. albicans in each well. If the well is turbid, the compound at that concentration has no anti-C. albicans activity. If the next well is visibly clear, the compound at that concentration has anti-C. albicans activity. This is the minimum inhibitory concentration (MIC) of the compound against C. albicans. The results are shown in Table 1.

[0306] Table 1

[0307]

[0308] As shown in Table 1, the 10,13-disubstituted berberine derivatives provided by the present invention have antifungal effects.

[0309] CLSM (Confocal Laser Scanning Microscopy) was used to observe the effects of berberine and its derivatives on the thickness of Candida albicans biofilms. Confocal laser scanning microscopy visually examined the inhibitory effects of drugs on biofilm formation. FUNXite-1 binds to the cell membrane and is converted by fungal cells to produce green fluorescence. The biofilm formed in the control group had a dense three-dimensional structure, with vigorous bacterial growth and a thicker biofilm. Figure 1 , showing that after drug intervention, the thickness of the biofilm of Candida albicans was significantly reduced and the amount of bacteria was reduced, indicating that berberine can inhibit the biofilm formation of Candida albicans; and berberine derivative 23 is more effective than berberine and has a stronger inhibitory effect.

[0310] Example 39 Effects of 10- and 13-position disubstituted berberine derivatives on mice with oropharyngeal candidiasis

[0311] The effects of the compounds of the present invention on oropharyngeal candidiasis were evaluated using a mouse model of oropharyngeal candidiasis (primarily based on the literature Solis NV, Filler SG. Mouse model of oropharyngeal candidiasis[J]. Nat Protoc, 2012, 7(4): 637-42). Treatment was performed at a dose gradient of 50 mg / kg, 100 mg / kg, and 200 mg / kg, with 100 mg / kg being the optimal treatment, and this concentration was used for subsequent experiments.

[0312] (1) Effects on tongue mucosa

[0313] The surface of the tongue mucosa of mice with oropharyngeal candidiasis was observed. Figure 2As shown in Table 1, it is shown that the tongue of the control group (without drug treatment) is smooth and moist, and the surface of the tongue mucosa is light red; after Candida albicans infection, the surface of the tongue mucosa is covered with a white pseudomembrane, and the overall structure is severely damaged. After drug (BBR or compound 23) intervention, the tongue mucosa of the mouse with oropharyngeal candidiasis is repaired to a certain extent, and the recovery effect after treatment with berberine derivative 23 is better than that after treatment with berberine.

[0314] (2) Oral fungal load test

[0315] The oral fungal load of the mouse with oropharyngeal candidiasis was determined. The Candida albicans load of the mouse was collected and plated, and the results are shown in Table 2. Figure 3 As shown in Table 2, it is shown that after drug treatment, the Candida albicans load shows a gradually decreasing trend, and with stable intervention of the drug, the proliferation of Candida albicans can be more effectively inhibited, and the intervention effect of the berberine derivative 23 (compound 23) of the application is better than that of BBR.

[0316] (3) HE staining

[0317] The tongue tissue of the mouse with oropharyngeal candidiasis was subjected to HE staining pathological section. As shown in Table 3, Figure 4 the effect of the drug on the pathological morphology of the tongue tissue of the mouse was observed. The surface of the tongue mucosa of the uninfected control group mouse was arranged in order, and the surface of the tongue mucosa was intact. After Candida albicans infection, the overall structure was destroyed, and after drug (BBR or compound 23) treatment, the damage to the surface structure of the tongue mucosa was gradually repaired to different degrees, and the inflammatory cell infiltration was gradually reduced; and the berberine derivative 23 recovered the damage to a better extent.

[0318] In order to further observe the damage to the tongue tissue of the mouse and the repair after drug treatment, the effect of the drug on the tongue mucosa of the mouse with oropharyngeal candidiasis was observed by SEM. The results are shown in Table 4. Figure 5 As shown in Table 4, it is shown that the overall structure of the tongue tissue of the uninfected control group mouse is complete, and the structure is destroyed after Candida albicans infection. The surface of the tongue mucosa treated with berberine is more severely damaged, and the repair degree is lower; and after treatment with the berberine derivative 23, the filiform papillae on the surface of the tongue mucosa gradually stretch, indicating that the berberine derivative 23 is better than berberine.

[0319] (4) Effect on gene expression and protein expression

[0320] In order to observe the effect of the drug on the gene expression in the tongue tissue of the mouse, qRT-PCR experiment was carried out, and the gene expression in the tongue tissue of the mouse with oropharyngeal candidiasis was detected by qRT-PCR. The results are shown in Table 5. Figure 6 As shown in Table 5, it is shown that after drug intervention, the S100a8 gene expression is significantly reduced, and the effect of the berberine derivative 23 is better than that of berberine, and the difference is statistically significant.

[0321] To observe the effect of the drug on the protein expression in the tongue tissue of mice, the protein expression in the tongue tissue of the oropharyngeal candidiasis mice was detected by WB. The results, as shown in Figure 7 indicated that, after the drug intervention, the EGFR protein expression in the tongue tissue of the oropharyngeal candidiasis mice was significantly reduced, and the protein expression amount after the treatment of berberine derivative 23 was lower.

[0322] In the description of the present specification, the description of the terms "one embodiment", "some embodiments", "an example", "a specific example", "some embodiments" or "some examples" and the like means that the specific features, structures, materials or characteristics described in connection with the embodiment or example are included in at least one embodiment or example of the present application. In the present specification, the illustrative description of the above terms does not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described can be combined in any appropriate manner in any one or more embodiments or examples. Furthermore, the different embodiments or examples described in the present specification and the features of the different embodiments or examples can be combined and combined by those skilled in the art without contradiction.

[0323] Although the embodiments of the present application have been shown and described above, it can be understood that the above embodiments are exemplary and cannot be understood as limiting the present application, and those skilled in the art can make changes, modifications, replacements and variations to the above embodiments within the scope of the present application.

Claims

1. A compound, characterized in that The compound is a compound represented by general formula (I) or a pharmaceutically acceptable salt of the compound represented by general formula (I): R1 is selected from -(CH2) m R3, -(CH2) n -CN, unsubstituted C2-C 10 Alkenyl, unsubstituted C2-C 10 Alkynyl, through at least one R a Substituted C2-C 10 alkyl; R2 is selected from -(CH2) m R3, unsubstituted C2-C 10 Alkenyl, unsubstituted C2-C 10 Alkynyl; m=1, n=1, R3 is unsubstituted or substituted with at least one R b substituted phenyl; R a is -NH2; R b Selected from -NO2, C1 alkyl, or when R1 is selected from unsubstituted C2-C 10 Alkenyl or unsubstituted C2-C 10 In the case of alkynyl, R2 is selected from unsubstituted C1-C 10 alkyl.

2. A compound, characterized in that The compound is the following compound or a pharmaceutically acceptable salt of the following compound: 。 3. A drug, characterized in that The medicament comprises the compound according to claim 1 or 2, and is used for improving or treating Candida albicans infection.

4. Use of the compound according to claim 1 or 2 in the preparation of a medicament for improving or treating Candida albicans infection.

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