CN113549046A - Bisbecklonin S derivative and preparation method and application thereof - Google Patents

Abstract

The application provides a double bendyrin S derivative, a preparation method and an application thereof, wherein the derivative has a structure shown in a formula I:

R₁、R₂each independently selected from hydrogen, methyl, - (CH)₂)_nR₉、‑(C＝O)R₉；R₃Is H or- (CH)₂)_nR₉；R₅、R₆、R₇、R₈Each independently selected from hydrogen, hydroxy, methyl, halogen, nitro and amino; r₉Selected from amino and 5-6 membered nitrogen containing rings; the 5-6 membered nitrogen containing ring may optionally contain another heteroatom oxygen atom; the amino group and the 5-to 6-membered nitrogen containing ring may be unsubstituted or selected from C_1‑5Alkyl radical, C_1‑5Haloalkyl, hydroxy, phenyl, by C_1‑5Alkoxy-or halogen-substituted phenyl, R₁₀S(＝O)₂-substituted with at least one substituent of (a); r₁₀Is selected from C_1‑5Alkyl, 5-6 membered nitrogen containing ring; n is 1 or 2; wherein R is₁、R₂Not methyl at the same time.

Description

Bisbecklonin S derivative and preparation method and application thereof

Technical Field

The application relates to the technical field of medicines, in particular to a double bendyline S derivative and a preparation method and application thereof.

Background

The information in this background section is disclosed only to enhance understanding of the general background of the application and is not necessarily to be construed as an admission or any form of suggestion that this information forms the prior art that is already known to a person of ordinary skill in the art.

Inflammation is a defense response of the body to stimuli. Any factor that can cause tissue damage, such as biological, physical and chemical factors, can induce inflammation. Inflammation can be classified into acute inflammation and chronic inflammation according to duration. Acute inflammation is generally manifested only as a vascular response, which is a normal defense response of the body to stimuli from inside or outside the body, and is beneficial to the body. However, if the acute inflammatory response cannot achieve the effect of eliminating the stimulation, inflammatory factors related to the acute inflammatory response will be continuously present and continuously produced, such as interleukins, chemokines, cell adhesion molecules, and inflammation-related enzymes, and the like, resulting in the occurrence of chronic inflammation. In chronic inflammation, the persistent presence of inflammatory factors causes tissue damage, which in turn further initiates inflammation to form a vicious circle, thereby inducing various diseases and extremely harmful to the body.

Rheumatoid arthritis is a common acute or chronic inflammation of connective tissue that can recur and involve the heart. Clinically, it is characterized by joint and muscle migratory soreness and pain, and belongs to allergic diseases. According to statistics, half of people over 50 years old in China suffer from rheumatoid arthritis, about 85% of people over 65 years old suffer from rheumatoid arthritis, the number and proportion of people are still increasing, and the disease form is very severe. The rheumatic arthritis brings inconvenience to the movement of the patient and seriously affects the life quality of the patient. There are a variety of treatments for rheumatoid arthritis, among which anti-inflammatory drugs play an important role in their treatment. Therefore, the discovery and development of anti-inflammatory drugs are of great significance for the treatment of rheumatoid arthritis.

In addition, inflammation is also closely linked to the development of tumors, and it is estimated that about 15% of cancers are caused by chronic inflammation, which plays an important role in the development and progression of tumors. Therefore, inhibition of inflammation is an effective measure for prevention and treatment of tumors.

The bibenzyl is a specific natural polyphenol compound in bryophytes, has wide and obvious biological activities of resisting fungi, microbes, oxidation, cytotoxicity, insect food refusal, plant growth regulation, platelet aggregation and the like, and is an important source for discovering and developing new medicines.

Disclosure of Invention

The cyclic bibenzyl compound with a brand new structure is obtained by separating in the herb of Reboulia hemisphaera (Reboulia hemisperica) for the first time by the inventor, is colorless blocky crystals, and has a molecular formula of C₂₉H₂₆O₅The molecular weight is 454, and the compound is easily dissolved in organic solvents such as chloroform, ethyl acetate and the like, and the structural formula is as follows:

the inventor names the livetin S (Marchantin S) and carries out biological activity research on the livetin S, and the result shows that the livetin S has good anti-inflammatory and anti-tumor activity, can reduce proinflammatory factors of tumor cells and induce autophagic apoptosis and senescence of the tumor cells, can become a novel anti-tumor drug or anti-inflammatory drug, has quite wide research prospect and development potential, further provides a total synthesis method of the livetin S, carries out structure-effect research on the livetin S, provides a series of compounds with anti-inflammatory and anti-tumor activity, and has very important significance for finding efficient anti-inflammatory and anti-tumor drugs.

Specifically, the technical scheme of the invention is as follows:

in a first aspect of the invention, the invention provides a dibenzj S derivative and pharmaceutically acceptable salts thereof, the derivative having a structure represented by formula I:

R₁、R₂each independently selected from hydrogen, methyl, - (CH)₂)_nR₉、-(C＝O)R₉；

R₃Is H or- (CH)₂)_nR₉；

R₅、R₆、R₇、R₈Each independently selected from hydrogen, hydroxy, methyl, halogen, nitro and amino;

R₉selected from amino and 5-6 membered nitrogen containing rings; the 5-6 membered nitrogen containing ring may optionally contain an additional heteroatom oxygen or nitrogen atom; the amino group and the 5-to 6-membered nitrogen containing ring may be unsubstituted or selected from C_1-5Alkyl radical, C_1-5Haloalkyl, hydroxy, phenyl, by C_1-5Alkoxy-or halogen-substituted phenyl, R₁₀S(＝O)₂-substituted with at least one substituent of (a);

R₁₀is selected from C_1-5Alkyl, 5-6 membered nitrogen containing ring;

n is 1 or 2;

wherein R is₁、R₂Not methyl at the same time.

Further, in an embodiment of the present invention, the derivative has a structure represented by formula II, formula III, formula IV or formula V below:

wherein in the structure of formula III, R₁Selected from hydrogen, - (CH)₂)_nR₉、-(C＝O)R₉；

In the structures of formula III and formula IV, R₂Is selected from- (CH)₂)_nR₉、-(C＝O)R₉；

In the structure of formula II, R₃Is- (CH)₂)_nR₉；

R₉And n is as defined above.

In an embodiment of the invention, the inventors have found that compounds having anti-tumor and/or anti-inflammatory activity when they conform to the structure shown in formula II, formula III, formula IV or formula V.

Further, in an embodiment of the present invention, R₉Selected from amino and a 5-6 membered nitrogen containing ring, said 5-6 membered nitrogen containing ring selected from morpholinyl, piperazinyl, piperidinyl and pyrrolidinyl; the amino group and the 5-to 6-membered nitrogen containing ring may be unsubstituted or selected from methyl, ethyl, C_1-2Haloalkyl, hydroxy, phenyl, methoxyphenyl, halophenyl, methylsulfonyl, or morpholinosulfonyl.

In particular, in embodiments of the present invention, the inventors have found that when the compounds of the present invention have the following structure (formula VI, VII or VIII), they tend to have particularly excellent anti-tumor and/or anti-inflammatory activity:

wherein ring A is a nitrogen-containing aliphatic ring, and ring A is selected from morpholinyl, piperazinyl, piperidinyl, and pyrrolidinyl;

x is selected from C, N or O;

R₁₁selected from hydrogen, methyl, hydroxy, phenyl, methoxyphenyl, halophenyl, methylsulfonyl, morpholinosulfonyl;

R₁₂、R₁₃each independently selected from C_1-5Alkyl or C_1-5Haloalkyl, preferably selected from methyl, and haloethyl.

In particular, when ring a is morpholinyl, piperazinyl or piperidinyl, i.e. the compound has the structure shown in formula VII (X is selected from C, N or O), it tends to have particularly excellent anti-inflammatory activity.

Especially, when R is₁₂And R₁₃When not identical, the compound has better activity.

In some embodiments of the invention, when ring A is selected from morpholinyl, piperidinyl, and pyrrolidinyl in the structure of formula VI, R is₁₁Is hydrogen; when ring A is piperazinyl, R₁₁Selected from methyl, hydroxy, phenyl, methoxyphenyl, halophenyl, methylsulfonyl, morpholinosulfonyl; preferably, R₁₁The substitution position is the N-terminal of the piperazinyl.

Further, in the above compound of formula III, R₁And R₂Are the same as, and are selected from- (CH)₂)_nR₉Or- (C ═ O) R₉The compound conforms to the structure of formula IX or formula X:

wherein R is₉Selected from morpholinyl, piperidinyl and pyrrolidinyl.

Further, in the above compound of formula IV, R₂Is selected from- (CH)₂)_nR₉、-(C＝O)R₉，R₉Selected from morpholinyl, piperazinyl, piperidinyl and pyrrolidinyl.

In an embodiment of the invention, R₉When the nitrogen-containing ring is a 5-6-membered nitrogen-containing ring, the nitrogen end of the nitrogen-containing ring is used as a connecting site to be connected with methylene or carbonyl, and the compound with the connecting characteristic under the general structure has remarkably better activity.

Further, by way of example, the present invention provides a series of compounds, as shown below:

in an embodiment of the present invention, the term "salt" refers to the above-mentionedAcid and/or base salts of the compounds or stereoisomers thereof, with inorganic and/or organic acids and bases also include zwitterionic (inner) salts and also quaternary ammonium salts, for example alkylammonium salts. These salts can be obtained directly in the final isolation and purification of the compounds. The compound or a stereoisomer thereof may be obtained by appropriately (e.g., equivalently) mixing the above compound or a stereoisomer thereof with a predetermined amount of an acid or a base. These salts may form precipitates in the solution which are collected by filtration, or they may be recovered after evaporation of the solvent, or they may be prepared by reaction in an aqueous medium followed by lyophilization. For example, in some embodiments, the salt of the present invention can be a potassium salt of the compound (KOH, K)₂CO₃) Sodium salt (NaOH, Na)₂CO₃) Calcium salt (CaCl)₂,Ca(OAc)₂) And magnesium salts (MgCl)₂) Etc.; or the pharmaceutically acceptable salt of the compound is a salt formed by the compound and inorganic acid (hydrochloric acid, sulfuric acid and hydrobromic acid) or organic acid (methanesulfonic acid, toluenesulfonic acid and trifluoroacetic acid).

In a second aspect of the present invention, the present invention provides a process for preparing the dibenzylidins derivative described in the first aspect above and pharmaceutically acceptable salts thereof, comprising the following reaction scheme:

scheme 1:

wherein R is₁、R₂、R₃、R₅、R₆、R₇、R₈As defined above.

Further, the process further comprises carrying out the following reaction scheme 2 or 3 starting from compound 18:

scheme 2:

scheme 3:

wherein R is₁、R₂、R₃、R₅、R₆、R₇、R₈、R₉、R₁₁、R₁₂、R₁₃As defined above.

In some embodiments of the invention, R₁、R₂Selected from methyl and hydrogen atoms, R₅、R₆、R₇And R₈Each independently selected from hydrogen, hydroxy, alkoxy, scheme 1 involves the following steps:

step a: stirring and heating the compound 1 and the compound 2, cuprous oxide and potassium carbonate in pyridine, filtering, evaporating the solvent after the reaction is finished, and purifying by silica gel column chromatography to obtain a compound 3;

step b: slowly adding sodium borohydride into an ethanol or tetrahydrofuran solution of the compound 3 under the ice-water bath condition, magnetically stirring at room temperature, evaporating the solvent after the reaction is finished, and purifying by silica gel column chromatography to obtain a compound 4;

step c: dissolving the compound 4 in acetonitrile, adding triphenylphosphine hydrobromide, stirring, heating and refluxing, and after the reaction is finished, evaporating the solvent to obtain a compound 5;

step d: dissolving a compound 6 and a compound 7 in a DMF solution, adding potassium carbonate, heating, stirring, after the reaction is finished, evaporating to remove the solvent, and purifying by silica gel column chromatography to obtain a compound 8;

step e: dissolving the compound 5 and the compound 8 in a dichloromethane solution, adding potassium carbonate and a small amount of 18-crown-6, heating, refluxing, stirring, evaporating to remove the solvent after the reaction is finished, and purifying by silica gel column chromatography to obtain a compound 9;

step f: dissolving the compound 9 in ethyl acetate or methanol solution, adding palladium carbon, stirring under hydrogen conditions, evaporating to remove the solvent after the reaction is finished, and purifying by silica gel column chromatography to obtain a compound 10;

step g: dissolving the compound 10 and isoamyl nitrite in an ether or tetrahydrofuran solution, heating, refluxing, stirring, evaporating to remove the solvent after the reaction is finished, and purifying by silica gel column chromatography to obtain a compound 11;

step h: slowly dripping the compound 11 into diethyl ether or tetrahydrofuran solution containing lithium aluminum hydride, heating for refluxing, stirring, after the reaction is finished, slowly adding an ammonium chloride saturated solution, extracting by dichloromethane, drying an organic phase by using sodium sulfate, filtering, evaporating to remove a solvent, and purifying by silica gel column chromatography to obtain a compound 12;

step i: dissolving the compound 12 in an ethanol or tetrahydrofuran solution of an inorganic acid, stirring, evaporating to remove the solvent after the reaction is finished, and purifying by silica gel column chromatography to obtain a compound 13;

step j: dissolving the compound 13, potassium carbonate and benzyl bromide in acetonitrile or DMF solution, stirring, heating, after the reaction is finished, carrying out suction filtration, evaporating to remove the solvent, and purifying by silica gel column chromatography to obtain a compound 14;

step k: dissolving the compound 14, thionyl chloride and triethylamine in a dichloromethane solution, stirring, evaporating to remove the solvent after the reaction is finished, and purifying by silica gel column chromatography to obtain a compound 15;

step l: dissolving the compound 15 in acetonitrile, adding triphenylphosphine, stirring, heating, and after the reaction is finished, evaporating the solvent to obtain a compound 16;

m: slowly dripping a dichloromethane solution of the compound 16 into a dichloromethane solution of sodium methoxide or sodium tert-butoxide, stirring, evaporating to remove the solvent after the reaction is finished, and purifying by silica gel column chromatography to obtain a compound 17;

step n: dissolving the compound 17 in ethyl acetate or methanol solution, adding palladium carbon, stirring under hydrogen conditions, after the reaction is finished, carrying out suction filtration, evaporating to remove the solvent, and purifying by silica gel column chromatography to obtain a compound 18.

Further, scheme 2 or 3 is performed after scheme 1 is finished, starting with product compound 18 of scheme 1, wherein the steps of scheme 2 comprise: dissolving the compound 18, potassium carbonate and methyl iodide in an acetone solution, stirring, heating, after the reaction is finished, carrying out suction filtration, evaporating to remove the solvent, and purifying by silica gel column chromatography to obtain a compound 19; dropwise adding boron tribromide into a dichloromethane solution of a compound 19 under a low-temperature condition, stirring, adding a sodium bicarbonate aqueous solution after the reaction is finished, extracting with dichloromethane, combining organic phases, drying with sodium sulfate, filtering, evaporating to remove a solvent, and purifying by silica gel column chromatography to obtain a compound 21;

wherein the steps of scheme 3 comprise: mixing compound 18 and secondary amine compound R with different substituents₉H or R₉Dissolving Cl and formaldehyde into dioxane solution, stirring, heating, evaporating solvent after reaction, and purifying by silica gel column chromatography to obtain compound 22 or 27. Wherein R is₁、R₂Is hydrogen, R₃As defined above.

Alternatively, in some embodiments of the invention, R₁、R₂Selected from hydrogen, - (CH)₂)_nR₉、-(C＝O)R₉，R₅、R₆、R₇And R₈Scheme 4, when each is independently selected from hydrogen, hydroxy, alkoxy, starting from compound 18:

wherein R is₉As defined hereinbefore; scheme 4 involves the following steps: dissolving a compound 18, chloroalkane hydrochloride with different substituted amino groups and potassium carbonate into an acetone solution, stirring, heating, filtering after the reaction is finished, evaporating the solvent, and purifying by silica gel column chromatography to obtain a series of compounds 21 f-k respectively;

dissolving the compound 18, morpholine formyl chloride, triethylamine and DMAP into a dichloromethane solution, stirring, heating, evaporating the solvent after the reaction is finished, and purifying by silica gel column chromatography to obtain compounds 21d and 21e respectively.

In some embodiments of the present invention, the present invention provides a total synthesis of marchantin S comprising performing the synthesis according to the following reaction scheme:

the compounds of the present invention can be prepared by a person skilled in the art with limited experimentation based on the reaction schemes provided by the present invention. It will be appreciated that, given the disclosure of the reaction schemes, the selection of suitable solvents and the achievement of further improved yields and purities are reasonably routine endeavors in the art, and such pursuits can be achieved through a limited number of experimental attempts, given the knowledge of the routine skill in the art.

In a third aspect of the invention, the invention provides a pharmaceutical composition comprising the bimatoxylin S derivative described in the first aspect above and a pharmaceutically acceptable salt thereof.

The invention also provides a pharmaceutical preparation, which comprises the double bendyline S derivative and the pharmaceutically acceptable salt thereof in the first aspect, and at least one pharmaceutically acceptable auxiliary material or pharmaceutical carrier.

The pharmaceutical composition or the pharmaceutical preparation can be a solid oral preparation, a liquid oral preparation or an injection, and can be tablets, dispersible tablets, enteric-coated tablets, chewable tablets, orally disintegrating tablets, capsules, sugar-coated agents, granules, dry powders, oral solutions, small water injection for injection, freeze-dried powder injection for injection, large infusion solutions or small infusion solutions.

"pharmaceutically acceptable adjuvant" refers to an ingredient of a pharmaceutical composition other than an active ingredient, which is not toxic to a subject. Such as excipients, pharmaceutically acceptable excipients including, but not limited to, buffers, stabilizers, preservatives, and the like. The pharmaceutically acceptable carriers and excipients of the present invention are generally well known to those skilled in the art or can be determined by those skilled in the art based on the actual circumstances.

By "pharmaceutical carrier" is meant a pharmaceutically acceptable solvent, suspending agent or carrier for the delivery of the compound into an animal or human. The carrier may be a liquid or solid and is selected according to the intended mode of administration. Proteins and liposomes are also drug carriers. The carrier may be present in the pharmaceutical composition in an amount of 1% to 98% by weight, usually about 80% by weight.

Some embodiments of the invention include methods of producing a pharmaceutical composition or pharmaceutical formulation comprising admixing at least one compound disclosed herein with a pharmaceutically acceptable adjuvant or carrier. The formulations are prepared by any suitable method, generally by uniformly mixing the active compound with liquid and/or finely divided solid excipients in the desired ratio, and then, if desired, shaping the resulting mixture into the desired shape. The compounds of the present invention may be formulated into pharmaceutical compositions or formulations using techniques well known to those skilled in the art. For example, the pharmaceutical preparation may be prepared according to the modern pharmaceutical preparation series compiled by Shenyang pharmaceutical university. In addition to those mentioned herein, suitable pharmaceutical excipients are known in the art, see for example the 2005 edition handbook of pharmaceutical excipients (fourth edition of original works), authors (en) r.c. lo (raymon dcrowe) (usa) p.j. seskiki (paul jsheskey).

In a fourth aspect of the present invention, the present invention provides the use of the double marchantin S derivative and its pharmaceutically acceptable salt in the first aspect, or the pharmaceutical composition or pharmaceutical preparation in the third aspect for preparing an anti-tumor drug and/or anti-inflammatory drug.

In some embodiments of the invention, the tumor of the invention includes, but is not limited to, liver cancer, non-small cell lung cancer, breast cancer, prostate cancer, bronchial epithelial-like cell-like cancer.

In some embodiments of the invention, the anti-inflammatory agent of the invention exhibits an inhibitory effect on the transcription level (gene expression level) of the proinflammatory factor interleukin-1 beta (IL1 beta) and/or the transcription level (gene expression level) of the proinflammatory factor interleukin-6 (IL 6).

In a fifth aspect of the present invention, the present invention provides the use of the double marchanol S derivative and the pharmaceutically acceptable salt thereof described in the first aspect above, or the pharmaceutical composition or the pharmaceutical preparation of the third aspect above, in the preparation of a medicament for treating and/or alleviating rheumatoid arthritis or a medicament for treating and/or alleviating tumors caused by inflammation.

In a sixth aspect of the present invention, the present invention provides the use of compound 20 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition or pharmaceutical preparation comprising the compound or the salt thereof, for the manufacture of an anti-inflammatory medicament, or for the manufacture of a medicament for the treatment and/or alleviation of rheumatoid arthritis and/or the alleviation of inflammation-induced tumors;

in a sixth aspect of the invention, the invention provides a method for treating a disease, comprising administering to a subject an effective amount of the dibenzylidins derivative according to the first aspect of the invention described above and pharmaceutically acceptable salts thereof, or the pharmaceutical composition or pharmaceutical preparation or compound 20 according to the third aspect described above and compositions or pharmaceutical preparations comprising compound 20.

The diseases include liver cancer, non-small cell lung cancer, breast cancer, prostate cancer, bronchial epithelial-like cell cancers, as well as rheumatoid arthritis and tumors caused by inflammation.

As used herein, "treatment" refers to clinical intervention in an attempt to alter the natural course of the individual being treated, and may be for the purpose of prophylaxis or in the course of clinical pathology. Desirable effects of treatment include, but are not limited to, preventing occurrence or recurrence of disease, alleviating symptoms, attenuating any direct or indirect pathological consequences of the disease, preventing metastasis, reducing the rate of disease progression, ameliorating or palliating the disease state, and alleviating or improving prognosis.

"subject" refers to an animal, preferably a mammal, most preferably a human, who has been the object of treatment, observation or experiment.

"therapeutically effective amount" means that amount of active compound or pharmaceutical agent, including a compound of the present invention, that elicits the biological or medicinal response in a tissue system, animal or human that is being sought by a researcher, veterinarian, medical doctor or other medical professional, which includes alleviation or partial alleviation of the symptoms of the disease, syndrome, condition or disorder being treated. It will be appreciated that the optimum dosage and interval for administration of a compound of the invention will be determined by the nature of the compound and external conditions, such as the form, route and site of administration and the particular mammal being treated, and that such optimum dosage may be determined by conventional techniques. It should also be recognized that the optimal course of treatment, i.e., the daily dosage of the compound over a nominal period of time, may be determined by methods known in the art.

Compared with the prior art, the invention has the advantages that: the invention relates to a cyclic bibenzyl compound liverwortmin S (Marchantin S) with a brand-new structure, which is separated from liverwortmin (Reboulia hemisphaerica) for the first time, and experiments show that the liverwortmin S has excellent anti-tumor activity and anti-inflammatory activity.

Detailed Description

The present application is further illustrated with reference to specific examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present application. The experimental procedures, in which specific conditions are not noted in the following examples, are generally carried out according to conventional conditions or according to conditions recommended by the manufacturers.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. The reagents or starting materials used in the present application can be purchased from conventional sources, and unless otherwise specified, the reagents or starting materials used in the present application can be used in the conventional manner in the art or in the product specification. In addition, any methods and materials similar or equivalent to those described herein can be used in the methods of the present application. The preferred embodiments and materials described herein are intended to be exemplary only.

Example 1 Total Synthesis of marchantin S

The synthetic route is as follows:

step b: preparation of Compound 3

A mixture of compound 2' (80g), p-bromobenzaldehyde (70g), potassium carbonate (100g) and copper oxide (10g) was stirred under reflux in pyridine (250mL), after the reaction was complete, the pyridine was distilled off and the residue was extracted with dichloromethane. The solution was concentrated in vacuo and the residue was purified by silica gel column chromatography eluting with EtOAc-PE solution to give the product as a colorless oil.¹H NMR(400MHz,CDCl₃)δ9.88(s,1H),7.79(d,J＝8.7Hz,2H),7.35(dd,J＝8.5,1.8Hz,1H),7.25(d,J＝1.9Hz,1H),7.00(d,J＝8.5Hz,1H),6.97(d,J＝8.6Hz,2H),5.45(s,1H),4.23(dd,J＝10.9,4.9Hz,2H),3.95(td,J＝12.3,2.2Hz,2H),3.77(s,3H),2.18(qt,J＝12.5,5.0Hz,1H),1.42(d,J＝13.5Hz,1H).

Step c: preparation of Compound 4

Sodium borohydride (35g) was added to a solution of compound 3' (120g) in anhydrous ethanol (1200mL) at low temperature. The resulting mixture was stirred at room temperature for 1-3 h. The reaction was quenched by slowly adding water at 0 ℃ and the ethanol was distilled off. The resulting mixture was extracted with EtOAc and the organic phase was dried over sodium sulfate. Insoluble material was removed by filtration, the solution was concentrated in vacuo, and the residue was purified by column chromatography on silica eluting with EtOAc-PE to give the product as a white oil.¹H NMR(400MHz,CDCl₃)δ7.26–7.22(m,3H),7.10(d,J＝2.0Hz,1H),6.97(d,J＝8.4Hz,1H),6.90(d,J＝8.6Hz,2H),5.38(s,1H),4.57(s,2H),4.19(dd,J＝10.8,5.0Hz,2H),3.95–3.89(m,2H),3.80(s,3H),2.13(ddd,J＝12.5,7.9,5.0Hz,1H),1.42–1.35(m,1H).

Step d: preparation of Compound 5

Compound 4' (110g) and triphenylphosphine hydrobromide (150g) were dissolved in MeCN (1100mL), stirred at reflux for 1-3h, the solution was concentrated in vacuo and the residue was purified by column chromatography on silica eluting with MeOH-DCM to give a white solid, mp 131-.¹H NMR(400MHz,CDCl₃)δ7.72–7.64(m,10H),7.58(dd,J＝7.7,3.3Hz,5H),7.20–7.15(m,1H),7.01–6.98(m,1H),6.94(s,1H),6.92(d,J＝2.2Hz,1H),6.89(d,J＝8.5Hz,1H),6.63(d,J＝8.3Hz,2H),5.37(s,1H),5.36(s,1H),5.33(s,1H),4.14(dd,J＝11.3,4.4Hz,2H),3.93–3.85(m,2H),3.73(s,3H),2.15–2.05(m,1H),1.40–1.33(m,1H).

Step e: preparation of Compound 7

N, N-diisopropylethylamine (122g) was added dropwise to a solution of 2,3, 4-trihydroxybenzaldehyde (58.00g) in DCM (600 mL). The resulting mixture was stirred at room temperature for 0.5-2 h. Chloromethyl ethyl ether (85g) was slowly added to the mixture at 0 ℃ and the resulting mixture was stirred at room temperature for 12 h. The reaction mixture was washed with saturated brine, and the organic phase was dried over anhydrous sodium sulfate. Insoluble material was removed by filtration, the solution was concentrated in vacuo, and the residue was purified by silica gel column chromatography, eluting with EtOAc-PE solution, to give the product as a yellow oil.¹H NMR(400MHz,CDCl₃)δ11.23(s,1H),9.71(s,1H),7.23(d,J＝8.8Hz,1H),6.81(d,J＝8.8Hz,1H),5.29(s,2H),5.18(s,2H),3.88(q,J＝7.1Hz,2H),3.71(q,J＝7.0Hz,2H),1.20–1.18(m,3H),1.18–1.15(m,3H).

Step f: preparation of Compound 8

Methyl 5-fluoro-2-nitrobenzoate (52 g) was added to a solution of compound 7(67.00g) and potassium tert-butoxide (42g) in DMF (500 mL). The resulting mixture was stirred at 80 ℃ for 2-8 h. After addition of EtOAc (2000mL), the reaction mixture was washed with saturated brine (300 mL. times.3), and the organic phase was dried over anhydrous sodium sulfate. Insoluble material was removed by filtration, the solution was concentrated in vacuo, and the residue was purified by silica gel column chromatography, eluting with EtOAc-PE solution, to give the product as a yellow oil.¹H NMR(400MHz,CDCl3)δ10.01(s,1H),8.00(d,J＝9.0Hz,1H),7.72(d,J＝8.8Hz,1H),7.28(d,J＝8.9Hz,1H),7.07(d,J＝2.6Hz,1H),7.01(dd,J＝9.0,2.7Hz,1H),5.36(s,2H),5.10(s,2H),3.90(s,3H),3.77(q,J＝7.0Hz,2H),3.67(q,J＝7.1Hz,2H),1.25(t,J＝7.0Hz,3H),1.16(t,J＝7.1Hz,3H).

Step g: preparation of Compound 9

Potassium carbonate (22g) and a trace of 18-crown-6 were added to a solution of compound 5 '(55 g) and compound 8' (35g) in anhydrous DCM (1000mL) and the resulting mixture was stirred at reflux for 5-12 h. Insoluble material was removed by filtration, the solution was concentrated in vacuo, and the residue was purified by column chromatography on silica eluting with EtOAc-PE to give the cis-trans isomeric product as a yellow oil.

Step h: preparation of Compound 10

25% Pd/C (400mg) and triethylamine (100. mu.L) were added to a solution of compound 9' (1.5g) in EtOAc (60 mL). The suspension was stirred at room temperature for 2-8h with hydrogen. Insoluble material was removed by filtration, the solution was concentrated in vacuo, and the residue was purified by silica gel column chromatography, eluting with EtOAc-PE solution, to give a yellow oil.¹H NMR(400MHz,CDCl₃)δ7.31(d,J＝3.0Hz,1H),7.24(dd,J＝8.4,2.0Hz,1H),7.08(d,J＝2.0Hz,1H),7.00(d,J＝5.4Hz,1H),6.98(d,J＝2.3Hz,2H),6.96(d,J＝5.3Hz,1H),6.88(d,J＝8.6Hz,1H),6.84(dd,J＝8.9,3.0Hz,1H),6.81(d,J＝8.5Hz,2H),6.58(d,J＝8.9Hz,1H),5.45(s,2H),5.39(s,1H),5.24(s,2H),5.08(s,2H),4.20(dd,J＝10.8,4.9Hz,2H),3.93(dd,J＝12.3,2.3Hz,2H),3.82(s,3H),3.80(s,3H),3.79–3.74(m,2H),3.75–3.69(m,2H),2.73(s,4H),2.23–2.09(m,1H),1.42–1.36(m,1H),1.24(t,J＝7.1Hz,3H),1.17(t,J＝7.1Hz,3H).

Step i: preparation of Compound 11

A solution of compound 10' (11g) in anhydrous THF (20mL) was added dropwise to a solution of isoamylnitrite (5g) in anhydrous THF (10mL) at 60 ℃. The resulting mixture was stirred at reflux for 4-9 h. The solution was concentrated in vacuo and the residue was purified by silica gel column chromatography eluting with EtOAc-PE solution to give the product as a yellow oil.¹H NMR(400MHz,CDCl₃)δ7.67(d,J＝7.7Hz,1H),7.53–7.48(m,1H),7.32(t,J＝8.0Hz,1H),7.24(dd,J＝8.4,1.9Hz,1H),7.07(d,J＝2.0Hz,1H),7.03(d,J＝8.6Hz,1H),6.99(dd,J＝8.2,2.0Hz,1H),6.97(d,J＝3.9Hz,1H),6.95(d,J＝4.0Hz,2H),6.91(d,J＝8.6Hz,1H),6.80(d,J＝8.5Hz,2H),5.38(s,1H),5.25(s,2H),5.07(s,2H),4.20(dd,J＝10.8,4.9Hz,2H),3.93(dd,J＝12.3,2.3Hz,2H),3.87(s,3H),3.81(s,3H),3.76(q,J＝7.1Hz,2H),3.68(q,J＝7.1Hz,2H),2.73(d,J＝6.0Hz,4H),2.23–2.09(m,1H),1.42–1.36(m,1H),1.24(t,J＝7.0Hz,3H),1.15(t,J＝7.1Hz,3H).

Step j: preparation of Compound 12

A solution of compound 11' (7g) in anhydrous THF (20mL) was added dropwise to lithium aluminum hydride (1g) at room temperatureAnhydrous THF suspension (20 mL). The resulting mixture was stirred at reflux for 1-3h and then cooled to 0 ℃. Slowly adding water to quench and react to a colloid state. Filter through funnel and wash with EtOAc, concentrate the solution in vacuo, purify the residue by column chromatography on silica eluting with EtOAc-PE solution to give the product as a colorless oil.¹H NMR(400MHz,CDCl₃)δ7.25(d,J＝1.8Hz,1H),7.23–7.21(m,1H),7.07(d,J＝7.1Hz,1H),7.02(d,J＝8.5Hz,1H),6.98(s,1H),6.96(s,2H),6.94(s,1H),6.91(d,J＝8.7Hz,1H),6.80(s,1H),6.78(s,2H),6.76(d,J＝8.3Hz,1H),5.38(s,1H),5.24(s,2H),5.08(s,2H),4.60(s,2H),4.20(dd,J＝11.4,4.6Hz,2H),3.92(t,J＝11.9Hz,2H),3.82(s,3H),3.76(q,J＝7.1Hz,2H),3.70(q,J＝7.0Hz,2H),2.72(s,4H),2.22–2.11(m,1H),1.39(d,J＝13.5Hz,1H),1.25(t,J＝6.6Hz,3H),1.15(t,J＝7.1Hz,3H)。

Step k: preparation of Compound 13

Compound 12' (3.5g) was dissolved in ethanol (80mL) and hydrochloric acid solution (30mL), and the resulting mixture was stirred at room temperature for 3-6 h. Saturated sodium bicarbonate solution was added dropwise and the ethanol was distilled off in vacuo. The resulting mixture was extracted with DCM to collect the organic phase, which was dried over anhydrous sodium sulfate. Insoluble material was removed by filtration, the solution was concentrated in vacuo, and the residue was purified by silica gel column chromatography, eluting with EtOAc-PE solution, to give the product as a colorless oil.¹H NMR(400MHz,CDCl₃)δ9.69(s,1H),7.55(dd,J＝8.3,1.9Hz,1H),7.31(d,J＝1.9Hz,1H),7.20(t,J＝7.9Hz,1H),7.08(d,J＝8.3Hz,1H),6.97(d,J＝7.6Hz,1H),6.94(d,J＝8.5Hz,2H),6.88(d,J＝8.5Hz,2H),6.80–6.74(m,2H),6.69–6.65(m,2H),6.22(s,1H),5.62(s,1H),4.58(s,2H),3.99(s,3H),2.76(t,J＝7.2Hz,2H),2.63(t,J＝7.3Hz,2H)。

Step l: preparation of Compound 14

Potassium carbonate (2g) and benzyl bromide (2g) were added to a solution of compound 13' (1.5g) in acetonitrile (30mL), and the resulting mixture was stirred at reflux for 1-5 h. Insoluble material was removed by filtration, the solution was concentrated in vacuo, and the residue was purified by silica gel column chromatography, eluting with EtOAc-PE solution, to give the product as a colorless oil.¹H NMR(400MHz,CDCl₃)δ9.79(s,1H),7.61(d,J＝8.3Hz,1H),7.42(d,J＝7.3Hz,2H),7.37(d,J＝8.1Hz,2H),7.34(d,J＝4.3Hz,1H),7.31(d,J＝7.7Hz,1H),7.23(d,J＝8.2Hz,1H),7.22–7.14(m,5H),7.08(d,J＝8.3Hz,1H),7.03(s,1H),7.01(s,1H),6.97(d,J＝7.4Hz,1H),6.88–6.83(m,5H),6.80(d,J＝7.8Hz,1H),5.11(s,2H),4.94(s,2H),4.59(d,J＝4.8Hz,2H),3.95(s,3H),2.80(s,4H)。

Step m: preparation of Compound 15

Thionyl chloride (250mg) and a trace amount of triethylamine were added to a solution of compound 14' (680mg) in DCM (20mL) at low temperature, stirred at room temperature for 2-8h, and the solvent was evaporated. The resulting colorless oily product and triphenylphosphine (400mg) were dissolved in acetonitrile (20mL), stirred at reflux for 10-30h, the solution was concentrated in vacuo and chromatographed on silica gel eluting with MeOH-DCM to give a white solid, mp 104-.¹H NMR(400MHz,CDCl₃)δ9.76(s,1H),7.64–7.57(m,10H),7.47(dd,J＝7.5,3.3Hz,5H),7.42(d,J＝7.3Hz,3H),7.36(dd,J＝14.2,6.5Hz,4H),7.20(q,J＝6.2Hz,3H),7.14(d,J＝6.6Hz,2H),7.09(d,J＝8.4Hz,1H),7.04(d,J＝8.0Hz,1H),6.98(d,J＝8.3Hz,2H),6.85(d,J＝8.3Hz,3H),6.80(dd,J＝14.2,5.8Hz,3H),6.32(s,1H),5.24(d,J＝14.5Hz,2H),5.08(s,2H),4.80(s,2H),3.93(s,3H),2.72–2.66(m,2H),2.61–2.55(m,2H)。

Step o: preparation of Compound 16

A solution of compound 15' (1.3g) in anhydrous DCM (200mL) was slowly added dropwise to a suspension of sodium methoxide (1g) in anhydrous DCM (150mL) and stirred for 6-24 h. Filtering, vacuum concentrating, purifying by silica gel column chromatography, eluting with DCM-PE solution to obtain cis-trans isomeric white solid.

Step p: compound 18' (ii)Liverwort extract S) Preparation of

Pd/C (200mg) was added to a solution of compound 16' (800mg) in EtOAc (30 mL). The suspension was stirred at room temperature for 1-4h with hydrogen. Insoluble material was removed by filtration, the solution was concentrated in vacuo and the residue was purified by column chromatography on silica eluting with EtOAc-PE to give a white solid, mp 83-84 ℃.¹H NMR(400MHz,CDCl₃)δ6.98(t,J＝7.8Hz,1H),6.93(d,J＝6.9Hz,2H),6.91(s,1H),6.88(s,1H),6.85(d,J＝6.5Hz,1H),6.78(d,J＝8.1Hz,1H),6.65(s,1H),6.61(d,J＝7.6Hz,2H),6.55(d,J＝8.2Hz,1H),6.35(d,J＝7.4Hz,1H),5.51(s,1H),4.72(s,1H),3.90(s,3H),2.97(d,J＝7.3Hz,2H),2.93(d,J＝7.5Hz,2H),2.90–2.81(m,2H),2.82–2.73(m,2H)。

EXAMPLE 2 Synthesis of Compounds 20 and 21a to 21c

Preparation of compound 20:

under low temperature conditions, 1M BBr₃A solution of (300mg) in anhydrous DCM (1.2mL) was added slowly and dropwise to a solution of marchantin S (compound 18') (70mg) in anhydrous DCM, and the reaction mixture was stirred for 1-2 h. The reaction was quenched by dropwise addition of saturated sodium bicarbonate solution, the reaction mixture was washed with saturated brine, and the organic phase was dried over anhydrous sodium sulfate. Filtration and concentration of the solution in vacuo, purification by column chromatography on silica eluting with EtOAc-PE solution afforded a white solid, mp 79-80 ℃.¹H NMR(400MHz,CDCl₃)δ6.99(d,J＝7.9Hz,1H),6.96(s,1H),6.93(d,J＝7.4Hz,2H),6.90–6.85(m,2H),6.74(dd,J＝8.1,1.9Hz,1H),6.64–6.62(m,1H),6.60(d,J＝8.4Hz,2H),6.55(dd,J＝8.2,2.1Hz,1H),6.38(d,J＝7.6Hz,1H),5.55(s,1H),5.54(d,J＝1.9Hz,1H),5.40(s,1H),4.84(s,1H),2.99(dd,J＝6.7,3.7Hz,2H),2.93(dd,J＝6.7,3.8Hz,2H),2.89–2.83(m,2H),2.81–2.75(m,2H)。

Preparation of Compounds 21a to 21 c:

potassium carbonate (70mg) and methyl iodide (50mg) were added to a solution of marchantin S (80mg) in acetone (15mL), and the resulting mixture was stirred under reflux for 6 h. Filtration and concentration of the solution in vacuo, purification by column chromatography on silica eluting with EtOAc-PE solution afforded compounds 21a, 21b, 21c as three white solids.

The compound 21a, mp 168-169 ℃ C,¹H NMR(400MHz,CDCl₃)δ7.09(d,J＝8.5Hz,1H),6.94(d,J＝5.9Hz,1H),6.92(d,J＝6.7Hz,2H),6.88(d,J＝8.5Hz,1H),6.84(d,J＝8.2Hz,1H),6.77(dd,J＝8.2,1.9Hz,1H),6.64–6.62(m,1H),6.61(s,1H),6.59(s,1H),6.51(dd,J＝8.2,2.0Hz,1H),6.30(d,J＝7.5Hz,1H),5.67(s,1H),5.52(d,J＝1.9Hz,1H),3.90(s,3H),3.61(s,3H),2.98(s,4H),2.84(dd,J＝8.3,3.3Hz,2H),2.80–2.75(m,2H).

compound 21b, mp 66-67 ℃.¹H NMR(600MHz,CDCl₃)δ6.95(d,J＝8.2Hz,2H),6.92(d,J＝5.6Hz,2H),6.84(d,J＝8.2Hz,1H),6.82(d,J＝8.5Hz,1H),6.77(d,J＝8.2Hz,1H),6.66(s,1H),6.61(d,J＝8.1Hz,2H),6.55–6.51(m,1H),6.28(d,J＝7.3Hz,1H),5.50(d,J＝8.8Hz,1H),5.37(s,1H),3.91(d,J＝9.7Hz,6H),2.98(d,J＝8.4Hz,4H),2.86–2.81(m,2H),2.77(d,J＝6.1Hz,2H).

Compound 21c, mp 67-68 ℃.¹H NMR(400MHz,CDCl₃)δ7.13(d,J＝8.6Hz,1H),6.91(d,J＝7.4Hz,2H),6.85(d,J＝2.5Hz,1H),6.83(d,J＝3.0Hz,1H),6.77(dd,J＝8.2,1.9Hz,1H),6.61(s,2H),6.59(s,1H),6.53(dd,J＝8.2,2.0Hz,1H),6.30(d,J＝7.5Hz,1H),5.57(d,J＝1.8Hz,1H),3.89(d,J＝6.1Hz,6H),3.59(s,3H),2.99(s,4H),2.83(dd,J＝8.2,3.3Hz,2H),2.76(dd,J＝8.4,3.4Hz,2H).

EXAMPLE 3 Synthesis of Compounds 21d to 21k

Preparation of compounds 21d and 21 e:

triethylamine (30mg) was added to a solution of marchantin S (50mg) in DCM (10 mL). The resulting mixture was stirred at room temperature for 5 min. To the mixture were added 4-morpholinocarbonyl chloride (25mg) and 4-dimethylaminopyridine (5mg), and the resulting mixture was stirred at room temperature for 1-3 h. The reaction was quenched by dropwise addition of water, and the resulting mixture was extracted with DCM to collect an organic phase, which was washed with saturated brine and dried over anhydrous sodium sulfate. Insoluble material was removed by filtration, the solution was concentrated in vacuo and the residue was purified by column chromatography on silica eluting with EtOAc-PE to give compounds 21d and 21e as two white solids.

Compound 21d, mp 100-.¹H NMR(600MHz,CDCl₃)δ7.24(d,J＝8.6Hz,1H),6.98(s,1H),6.93(s,1H),6.91(d,J＝6.6Hz,2H),6.83(d,J＝8.2Hz,1H),6.78–6.75(m,1H),6.57(d,J＝7.9Hz,2H),6.52(d,J＝6.9Hz,2H),6.27(d,J＝7.4Hz,1H),5.46–5.44(m,1H),3.89(s,3H),3.48–3.44(m,2H),3.32–3.28(m,2H),3.12–3.08(m,2H),2.99(s,4H),2.88–2.80(m,4H),2.71(s,2H).

Compound 21e, mp 172-173 ℃.¹H NMR(400MHz,CDCl₃)δ7.33(d,J＝8.6Hz,1H),7.14(d,J＝8.6Hz,1H),6.95(d,J＝8.3Hz,2H),6.91(t,J＝7.9Hz,1H),6.83(d,J＝8.2Hz,1H),6.76(dd,J＝8.2,1.8Hz,1H),6.59(d,J＝8.0Hz,3H),6.53(dd,J＝8.2,2.2Hz,1H),6.26(d,J＝7.4Hz,1H),5.47(d,J＝1.7Hz,1H),3.89(s,3H),3.74–3.62(m,4H),3.62–3.50(m,4H),3.46(s,2H),3.29(s,2H),3.14–2.97(m,6H),2.89–2.79(m,4H),2.72(d,J＝5.5Hz,2H).

Preparation of compounds 21f and 21 g:

potassium carbonate (50mg) and N- (2-chloroethyl) piperidine hydrochloride (30mg) were added to a solution of marchantin S (50mg, N) in acetone (10mL), and the resulting mixture was stirred at reflux for 6 h. Insoluble material was removed by filtration, the solution was concentrated in vacuo and the residue was purified by column chromatography on silica eluting with MeOH in DCM to afford compound 21f and 21g as two white solids.

Compound 21f, mp 163-164 ℃.¹H NMR(600MHz,CDCl₃)δ7.07(d,J＝8.6Hz,1H),6.95(d,J＝8.3Hz,2H),6.90(t,J＝7.9Hz,1H),6.86(d,J＝8.6Hz,1H),6.84(d,J＝8.4Hz,1H),6.77(dd,J＝8.2,1.8Hz,1H),6.65(s,1H),6.62(d,J＝8.2Hz,2H),6.48(dd,J＝8.2,2.1Hz,1H),6.24(d,J＝7.5Hz,1H),5.50(d,J＝1.8Hz,1H),3.90(s,3H),3.74–3.70(m,2H),2.99–2.95(m,2H),2.94–2.89(m,2H),2.84–2.80(m,2H),2.78–2.73(m,2H),2.58–2.31(m,6H),1.67(q,J＝5.5Hz,4H).

Compound 21g, mp 119-120 ℃.¹H NMR(600MHz,CDCl₃)δ7.15(d,J＝8.5Hz,1H),6.95(d,J＝7.6Hz,2H),6.89(d,J＝7.7Hz,1H),6.86(d,J＝7.9Hz,1H),6.83(d,J＝8.2Hz,1H),6.76(d,J＝8.2Hz,1H),6.62(d,J＝7.5Hz,2H),6.60(s,1H),6.45(d,J＝7.9Hz,1H),6.24(d,J＝7.2Hz,1H),5.45(s,1H),4.31(s,4H),3.90(s,3H),3.35(s,4H),3.02–2.80(m,8H),2.79–2.36(m,8H),1.62(d,J＝97.1Hz,12H).

Preparation of compounds 21h and 21 i:

reference compounds 21f and 21 g. The marchantin S and N- (2-chloroethyl) morpholine hydrochloride are used as raw materials to obtain two white solids of compounds 21h and 21 i.

Compound 21h, mp 78-79 ℃.¹H NMR(400MHz,CDCl₃)δ7.09(d,J＝8.5Hz,1H),6.97–6.92(m,2H),6.89(d,J＝7.9Hz,1H),6.84(dd,J＝8.4,3.1Hz,2H),6.76(dd,J＝8.2,1.9Hz,1H),6.62(d,J＝8.4Hz,3H),6.51–6.44(m,1H),6.25(d,J＝7.5Hz,1H),5.50(s,1H),3.90(s,3H),3.78(s,6H),3.00–2.95(m,2H),2.94–2.89(m,2H),2.82(dd,J＝7.9,3.3Hz,2H),2.78–2.71(m,2H),2.61–2.48(m,4H),2.48–2.41(m,2H).

The compound (21 i) is a compound represented by the formula,¹H NMR(600MHz,CDCl₃)δ7.10(d,J＝8.6Hz,1H),6.92(d,J＝8.2Hz,2H),6.87(t,J＝7.9Hz,1H),6.83(d,J＝8.4Hz,1H),6.81(d,J＝8.8Hz,1H),6.76(dd,J＝8.2,1.6Hz,1H),6.61(s,1H),6.60(s,2H),6.44(dd,J＝8.2,2.2Hz,1H),6.23(d,J＝7.5Hz,1H),5.51(d,J＝1.7Hz,1H),4.14(t,J＝5.9Hz,2H),3.92(t,J＝5.0Hz,2H),3.89(s,3H),3.74–3.69(m,4H),3.53(s,4H),2.96(s,4H),2.81(t,J＝5.8Hz,4H),2.76–2.70(m,2H),2.57(s,4H),2.38(s,2H),2.24(s,4H).

preparation of compounds 21j and 21 k:

reference compounds 21f and 21 g. The marchantin S and N- (2-chloroethyl) pyrrolidine hydrochloride are used as raw materials to obtain two white solids of compounds 21j and 21 k.

Compound 21j, mp 75-76 ℃.¹H NMR(400MHz,CDCl₃)δ7.07(d,J＝8.5Hz,1H),6.95(s,1H),6.93(s,1H),6.89(d,J＝7.9Hz,1H),6.85(s,1H),6.83(s,1H),6.76(dd,J＝8.2,1.9Hz,1H),6.63(d,J＝2.3Hz,2H),6.61(s,1H),6.49(dd,J＝8.1,2.2Hz,1H),6.24(d,J＝7.5Hz,1H),5.50(d,J＝1.9Hz,1H),3.90(s,3H),3.80–3.74(m,2H),2.99–2.94(m,2H),2.94–2.89(m,2H),2.82(dd,J＝8.1,3.2Hz,2H),2.78–2.72(m,2H),2.62(s,4H),2.60–2.54(m,2H),1.85(s,4H).

Compound 21k, mp 94-95 ℃.¹H NMR(600MHz,CD₃OD)δ7.31(d,J＝8.8Hz,1H),7.08(d,J＝8.8Hz,1H),6.98(d,J＝2.3Hz,1H),6.97–6.95(m,2H),6.92(d,J＝8.3Hz,1H),6.81(dd,J＝8.3,2.0Hz,1H),6.64(s,1H),6.54(d,J＝8.3Hz,2H),6.52(dd,J＝8.2,2.1Hz,1H),6.33(d,J＝7.5Hz,1H),5.56(d,J＝1.9Hz,1H),4.41–4.37(m,2H),4.11–4.08(m,2H),3.84(s,3H),3.55(s,2H),3.34(d,J＝15.7Hz,4H),3.06–3.04(m,2H),3.00(d,J＝9.2Hz,4H),2.97(s,4H),2.84(d,J＝5.7Hz,2H),2.77(d,J＝5.5Hz,2H),2.07(t,J＝6.6Hz,4H),1.89–1.85(m,4H).

EXAMPLE 4 Synthesis of Compounds 22 a-c, 27 a-27 g and 27 i-27 k

Preparation of compound 22 a:

marchantin S (0.10mmol) was dissolved in 1, 4-dioxane (10mL), 37% formaldehyde (0.15mmol) and N-phenylpiperazine (0.15mmol) were added sequentially, stirred at reflux for 12-24h, the solution was concentrated in vacuo, the residue was purified by silica gel column chromatography eluting with MeOH-DCM to give a brown solid, mp 109-.¹H NMR(400MHz,CDCl₃)δ7.24(s,1H),7.22(d,J＝4.8Hz,1H),6.90(dd,J＝9.7,4.3Hz,5H),6.85(d,J＝7.4Hz,1H),6.80(d,J＝8.1Hz,1H),6.73(d,J＝8.1Hz,1H),6.64(s,1H),6.59(d,J＝7.4Hz,2H),6.56(s,1H),6.52(d,J＝8.2Hz,1H),6.24(d,J＝7.3Hz,1H),5.50(s,1H),3.86(s,3H),3.80(s,2H),3.23(s,4H),2.91(s,4H),2.87–2.65(m,8H).

Preparation of compound 22 b:

the synthesis of reference compound 22a was carried out using marchantin S and 1- (4-methoxy) phenylpiperazine as starting materials to give a brown solid, mp 119-.¹H NMR(400MHz,CDCl₃)δ6.96–6.93(m,2H),6.92(dd,J＝5.7,3.2Hz,3H),6.86(d,J＝4.6Hz,2H),6.84(d,J＝3.6Hz,1H),6.77(dd,J＝8.2,1.9Hz,1H),6.68(s,1H),6.65(s,1H),6.61(d,J＝8.4Hz,2H),6.55(dd,J＝8.2,2.1Hz,1H),6.28(d,J＝7.5Hz,1H),5.52(d,J＝1.9Hz,1H),3.90(s,3H),3.86(s,2H),3.78(s,3H),3.19(s,4H),2.95(s,4H),2.89–2.74(m,8H).

Preparation of compound 22 c:

reference compound 22a was synthesized from marchantin S and 1- (4-bromo) phenylpiperazine as starting materials to give a yellow solid, mp 219-.¹H NMR(400MHz,CDCl₃)δ7.36(d,J＝8.8Hz,2H),6.95(d,J＝2.9Hz,1H),6.93(t,J＝3.7Hz,2H),6.85(d,J＝8.2Hz,1H),6.81(s,1H),6.78(d,J＝8.0Hz,2H),6.68(s,1H),6.63(d,J＝6.8Hz,2H),6.60(s,1H),6.56(dd,J＝8.3,2.4Hz,1H),6.28(d,J＝7.4Hz,1H),5.54–5.51(m,1H),3.91(s,3H),3.83(s,2H),3.23(s,4H),2.95(s,4H),2.90–2.66(m,8H).

Preparation of compound 27 a:

reference compound 22a was synthesized from marchantin S and dimethylamine as starting materials to give a brown solid, mp145-146 ℃.¹H NMR(400MHz,CDCl₃)δ6.97–6.92(m,2H),6.91(s,1H),6.84(d,J＝8.2Hz,1H),6.77(dd,J＝8.2,1.8Hz,1H),6.67(s,1H),6.61(d,J＝2.2Hz,2H),6.59(s,1H),6.57–6.52(m,1H),6.28(d,J＝7.4Hz,1H),5.53(d,J＝1.8Hz,1H),3.90(s,3H),3.76(s,2H),2.94(s,4H),2.83(dd,J＝8.3,3.3Hz,2H),2.76(dd,J＝8.2,3.0Hz,2H),2.43(s,6H).

Preparation of compound 27 b:

reference compound 22a was synthesized from marchantin S and diethylamine as starting materials to give a brown solid, mp111-112 ℃.¹H NMR(400MHz,CDCl₃)δ6.94(d,J＝3.1Hz,1H),6.91(d,J＝3.8Hz,1H),6.84(d,J＝8.2Hz,2H),6.79–6.74(m,1H),6.66(s,1H),6.61(d,J＝9.0Hz,2H),6.58(s,1H),6.57–6.53(m,1H),6.26(d,J＝7.3Hz,1H),5.52(d,J＝1.7Hz,1H),3.89(s,3H),3.88(s,2H),2.94(s,4H),2.82(d,J＝7.4Hz,2H),2.76(q,J＝7.0Hz,6H),1.19(t,J＝7.1Hz,6H).

Preparation of compound 27 c:

reference compound 22a was synthesized from marchantin S and 2-chloro-N-methylethylamine hydrochloride as starting materials to give a white solid, mp 64-65 ℃.¹H NMR(400MHz,CDCl₃)δ6.97(d,J＝6.6Hz,2H),6.94(s,1H),6.86(d,J＝8.2Hz,1H),6.79(dd,J＝8.2,1.7Hz,1H),6.69(s,1H),6.64(d,J＝3.6Hz,2H),6.61(s,1H),6.59–6.55(m,1H),6.30(d,J＝7.5Hz,1H),5.56(d,J＝1.8Hz,1H),3.92(s,3H),3.85(s,2H),3.73(t,J＝6.1Hz,2H),2.97(s,4H),2.95(d,J＝6.1Hz,2H),2.86(dd,J＝8.0,3.0Hz,2H),2.79(dd,J＝8.2,3.1Hz,2H),2.42(s,3H).

Preparation of compound 27 d:

reference compound 22a was synthesized from marchantin S and piperidine to give a white solid, mp184-185 ℃.¹H NMR(400MHz,CDCl₃)δ6.94(d,J＝3.7Hz,1H),6.91(d,J＝3.5Hz,1H),6.84(d,J＝8.2Hz,2H),6.77(dd,J＝8.2,1.9Hz,1H),6.68(s,1H),6.60(d,J＝8.4Hz,2H),6.58(s,1H),6.55(dd,J＝8.2,2.2Hz,1H),6.27(d,J＝7.5Hz,1H),5.53(d,J＝1.9Hz,1H),3.90(s,3H),3.75(s,2H),2.94(d,J＝8.0Hz,4H),2.89–2.40(m,8H),1.73–1.63(m,4H),1.63–1.44(m,2H).

Preparation of compound 27 e:

reference compound 22a was synthesized from marchantin S and tetrahydropyrrole as starting materials to give a yellow solid, mp208-209 ℃.¹H NMR(400MHz,CDCl₃)δ6.91(d,J＝8.1Hz,2H),6.85–6.81(m,2H),6.76(dd,J＝8.3,1.6Hz,1H),6.64(s,1H),6.58(d,J＝8.3Hz,2H),6.52(d,J＝7.9Hz,1H),6.45(d,J＝8.0Hz,1H),6.30–6.23(m,1H),5.55–5.49(m,1H),3.89(s,3H),3.84(s,2H),2.95(d,J＝18.5Hz,4H),2.85–2.70(m,4H),2.01(d,J＝23.8Hz,4H),1.25(t,J＝7.1Hz,4H).

Preparation of compound 27 f:

reference compound 22a was synthesized from marchantin S and N-methylpiperazine as starting materials to give a white solid, mp110-111 ℃.¹H NMR(400MHz,CDCl₃)δ6.94(d,J＝6.8Hz,2H),6.91(s,1H),6.84(d,J＝8.2Hz,1H),6.77(dd,J＝8.2,1.8Hz,1H),6.67(s,1H),6.63–6.60(m,2H),6.59(s,1H),6.54(dd,J＝8.2,2.1Hz,1H),6.28(d,J＝7.5Hz,1H),5.52(d,J＝1.8Hz,1H),3.90(s,3H),3.80(s,2H),3.00–2.60(m,16H),2.39(s,3H).

Preparation of compound 27 g:

of reference Compound 22aThe synthesis method takes the marchantin S and the morpholine as raw materials to obtain a white solid, and the temperature is mp203-204 ℃.¹H NMR(400MHz,CDCl₃)δ6.97–6.92(m,2H),6.92(s,1H),6.84(d,J＝8.2Hz,1H),6.77(dd,J＝8.2,1.7Hz,1H),6.67(s,1H),6.61(s,2H),6.59(s,1H),6.54(dd,J＝8.2,2.1Hz,1H),6.28(d,J＝7.5Hz,1H),5.53(d,J＝1.7Hz,1H),3.90(s,3H),3.77(s,6H),2.94(s,4H),2.83(dd,J＝8.4,3.0Hz,2H),2.77(d,J＝7.6Hz,2H),2.64(s,4H).

Preparation of compound 27 i:

reference compound 22a was synthesized from marchantin S and 4-hydroxypiperidine as starting materials to give a brown solid, mp118-119 ℃.¹H NMR(400MHz,CDCl₃)δ6.94(d,J＝7.8Hz,2H),6.91(s,1H),6.84(d,J＝8.2Hz,1H),6.77(dd,J＝8.2,1.8Hz,1H),6.67(s,1H),6.61(s,2H),6.59(s,1H),6.54(dd,J＝8.3,2.1Hz,1H),6.27(d,J＝7.5Hz,1H),5.52(d,J＝1.8Hz,1H),3.90(s,3H),3.81(s,2H),3.66–3.55(m,1H),2.94(s,6H),2.89–2.71(m,6H),2.06–1.98(m,2H),1.76–1.66(m,2H).

Preparation of compound 27 j:

the synthesis of reference compound 22a was carried out using marchantin S and N-methylsulfonylpiperazine as starting materials to give a white solid, mp 228-.¹H NMR(400MHz,CDCl₃)δ6.94(d,J＝8.6Hz,2H),6.91(s,1H),6.84(d,J＝8.2Hz,1H),6.77(dd,J＝8.2,1.7Hz,1H),6.67(s,1H),6.63(s,1H),6.60(d,J＝8.3Hz,2H),6.53(dd,J＝8.1,2.1Hz,1H),6.29(d,J＝7.5Hz,1H),5.52(d,J＝1.7Hz,1H),3.90(s,3H),3.83(s,2H),3.34(s,4H),2.94(s,4H),2.86–2.74(m,11H).

Preparation of compound 27 k:

the synthesis of reference compound 22a was carried out using marchantin S and morpholine N-methylsulfonylpiperazine as starting materials to give a white solid, mp 119-.¹H NMR(600MHz,CDCl₃)δ6.93(t,J＝8.9Hz,3H),6.84(d,J＝8.2Hz,1H),6.77(d,J＝8.1Hz,1H),6.68(s,1H),6.61(d,J＝6.9Hz,2H),6.59(s,1H),6.55–6.52(m,1H),6.27(d,J＝7.4Hz,1H),5.48(s,1H),3.90(s,3H),3.81(s,2H),3.76–3.72(m,4H),3.48–3.31(m,4H),3.28–3.22(m,4H),2.94(d,J＝8.8Hz,4H),2.85–2.80(m,2H),2.80–2.58(m,6H).

Experimental example 1 evaluation of antitumor Activity of Compound

Liverwort S and derivatives thereof (prepared according to the methods of examples 1-4) were assayed for their inhibitory effects on human liver cancer cells (HepG2), human non-small cell lung cancer cells (a549), human breast cancer cells (MCF-7), human prostate cancer cells (PC3) and human bronchial epithelial-like cells (HBE) using the MTT method.

The method comprises the following steps: MTT method: taking HepG2, A549, MCF-7, PC3 and HBE cell strains, 2 x 10⁴The cells/well are inoculated in a 96-well plate, 0.4% DMSO and compounds of different concentrations are respectively added, after 48h of culture, 5mg/mL MTT20 μ l is added into each well for further incubation for 4h, centrifugation and careful aspiration of supernatant are carried out, 200 μ l DMSO is added and slight shaking is carried out, after the generated formazan is completely dissolved and developed, the OD value is measured by using a Bio-Rad Model 550Microplate Reader at the wavelength of 570 nm. The experiments were performed 3 times under different incubation times, and the average was calculated, the specific results are shown in table 1.

TABLE 1 antitumor Activity of marchantin S and derivatives

The MTT screening result shows that: among phenolic hydroxyl modified derivatives, compound 20 showed significant inhibitory effect on 4 cancer cells, half of which inhibited concentration IC₅₀All are less than 12 mu M, have little influence on human normal cell HBE cells, and the compounds 21a, 21b and 21d have the surface effects on three cancer cells, namely HepG2, A549 and MCF-7Shows good inhibitory activity, has no obvious effect on HBE cells and PC3 cells, shows very obvious cytotoxicity on 5 cells of compounds 21g, 21j and 21k, particularly has very obvious effect on 21g and 21k, and has half inhibitory concentration IC₅₀Are all less than 3 μ M. Among aminomethylated modified derivatives, compound 27i showed inhibitory activity against HepG2 and a549 cells, and compound 27k showed inhibitory activity against HepG2 cells.

Experimental example 2 evaluation of anti-inflammatory Activity of Compound

The experimental method comprises the following steps: spreading inflammatory cells into a 6-well plate, adding 10 mu M of a solution of a compound to be detected to act on the cells for a certain time, then removing a culture medium, extracting total RNA of the cells by a Trizol method, carrying out reverse transcription into cDNA by a reverse transcription kit, carrying out PCR amplification, and calculating relative transcription levels (relative folds) by using software.

Anti-inflammatory results are shown in table 2.

TABLE 2 anti-inflammatory Activity of marchantin S and derivatives

^aN/T means have not been tested.

Results of transcription levels of proinflammatory factor interleukin-1 beta (IL1 beta) show that 8 compounds in the derivatives showed better inhibitory effects on transcription levels of IL1 beta than that of marchantin S, with compound 27c having the most significant inhibitory effect and 9 compounds having effects on transcription levels of IL1 beta comparable to that of marchantin S.

Results of the transcriptional level of the proinflammatory factor interleukin-6 (IL6) showed that 11 compounds of the derivatives showed better inhibitory effects on the transcriptional level of IL6 than marchantin S, with compounds 20 and 27d having the most significant inhibitory effects.

In conclusion, the marchantin S and the derivatives thereof can obviously inhibit the protein expression of inflammatory factors IL-6 and IL-1 beta, can reduce the gene expression level of the inflammatory factors such as IL-6 and IL-1 beta and the like, and show good anti-inflammatory effect. In addition, some of the derivatives showed good antitumor activity. The marchantin S and the derivatives thereof are novel medicaments with good development prospects for resisting inflammation and rheumatic arthritis and treating tumors caused by inflammation.

Although the present application has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that modifications may be made to the embodiments described in the foregoing embodiments, or equivalents may be substituted for elements thereof. Any modification, equivalent replacement, improvement and the like made within the spirit and principle of the present application shall be included in the protection scope of the present application.

Claims (10)

1. A dibenzmarchanol S derivative and pharmaceutically acceptable salts thereof, wherein the derivative has the structure shown in formula I:

R₁、R₂each independently selected from hydrogen, methyl, - (CH)₂)_nR₉、-(C＝O)R₉；

R₃Is H or- (CH)₂)_nR₉；

R₅、R₆、R₇、R₈Each independently selected from hydrogen, hydroxy, methyl, halogen, nitro and amino;

R₉selected from amino and 5-6 membered nitrogen containing rings; the 5-6 membered nitrogen containing ring may optionally contain another heteroatom oxygen atom; the amino group and the 5-to 6-membered nitrogen containing ring may be unsubstituted or selected from C_1-5Alkyl radical, C_1-5Haloalkyl, hydroxy, phenyl, by C_1-5Alkoxy-or halogen-substituted phenyl, R₁₀S(＝O)₂-substituted with at least one substituent of (a);

R₁₀is selected from C_1-5Alkyl, 5-6 membered nitrogen containing ring;

n is 1 or 2;

wherein R is₁、R₂Not methyl at the same time.

2. The dibenzmarchine S derivative of claim 1, and pharmaceutically acceptable salts thereof, having the structure of formula II or III:

wherein R is₁Selected from hydrogen, - (CH)₂)_nR₉、-(C＝O)R₉；

R₂Is selected from- (CH)₂)_nR₉、-(C＝O)R₉；

R₃Is- (CH)₂)_nR₉；

R₉And n is as defined in claim 1.

3. The dibenzmarchine S derivative of claim 1, and pharmaceutically acceptable salts thereof, wherein R is₉Selected from amino and a 5-6 membered nitrogen containing ring, said 5-6 membered nitrogen containing ring selected from morpholinyl, piperazinyl, piperidinyl and pyrrolidinyl; the amino group and the 5-to 6-membered nitrogen containing ring may be unsubstituted or selected from methyl, ethyl, C_1-2Substituted by at least one substituent selected from the group consisting of haloalkyl, hydroxy, phenyl, methoxyphenyl, halophenyl, methylsulfonyl, and morpholinosulfonyl;

preferably, the derivative corresponds to the structure of formula VI, VII or VIII:

wherein ring A is a nitrogen-containing aliphatic ring, and ring A is selected from morpholinyl, piperazinyl, piperidinyl, and pyrrolidinyl;

x is selected from C, N or O;

R₁₁selected from hydrogen, methyl, hydroxy, phenyl, methoxyphenyl, halophenyl, methylsulfonyl, morpholinosulfonyl;

R₁₂、R₁₃each independently selected from C_1-5Alkyl or C_1-5Haloalkyl, preferably selected from methyl, and haloethyl;

preferably, ring a is selected from morpholinyl, piperazinyl and piperidinyl;

preferably, R₁₂And R₁₃Different;

preferably, in the structure of formula VI, when ring A is selected from morpholinyl, piperidinyl and pyrrolidinyl, R₁₁Is hydrogen; when ring A is piperazinyl, R₁₁Selected from methyl, hydroxy, phenyl, methoxyphenyl, halophenyl, methylsulfonyl, morpholinosulfonyl; preferably, R₁₁The substitution position is the N-terminal of piperazinyl;

preferably, in the compound of formula III, R₁And R₂Are the same as, and are selected from- (CH)₂)_nR₉Or- (C ═ O) R₉The compound conforms to the structure of formula IX or formula X:

wherein R is₉Selected from morpholinyl, piperidinyl and pyrrolidinyl.

Preferably, in the compounds of formula IV, R₂Is selected from- (CH)₂)_nR₉、-(C＝O)R₉，R₉Selected from morpholinyl, piperazinyl, piperidinyl and pyrrolidinyl;

preferably, R₉In the case of a 5-to 6-membered nitrogen-containing ring, the nitrogen-containing ring is linked to a methylene group or a carbonyl group with its nitrogen terminal as a linking site.

4. The bimatorchidipine S derivative according to any one of claims 1 to 3, and pharmaceutically acceptable salts thereof, wherein said derivative is selected from the group consisting of:

5. a process for the preparation of the bimatorchinorin S derivative of any one of claims 1 to 4 and pharmaceutically acceptable salts thereof, according to the following reaction scheme:

wherein R is₁、R₂、R₃、R₅、R₆、R₇、R₈As defined in any one of claims 1 to 4.

Further, the process also includes the following reaction scheme starting with compound 18':

and/or the presence of a gas in the gas,

wherein R is₁、R₂、R₃、R₅、R₆、R₇、R₈、R₉、R₁₁、R₁₂、R₁₃As defined in any one of claims 1 to 4.

6. A pharmaceutical composition comprising the double marchanol S derivative of any one of claims 1 to 4 and pharmaceutically acceptable salts thereof.

7. A pharmaceutical formulation comprising the double marchanol S derivative of any one of claims 1 to 4 and pharmaceutically acceptable salts thereof and at least one pharmaceutically acceptable adjuvant or carrier.

8. Use of the dibenzj S derivative according to any one of claims 1 to 4 and pharmaceutically acceptable salts thereof, or the pharmaceutical composition according to claim 6 or the pharmaceutical preparation according to claim 7 for the preparation of an anti-tumor and/or anti-inflammatory agent;

preferably, the tumor includes, but is not limited to, liver cancer, non-small cell lung cancer, breast cancer, prostate cancer, bronchial epithelial cell-like cancers.

9. Use of the bimatoxylin S derivative according to any one of claims 1 to 4, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 6, or the pharmaceutical formulation according to claim 7, for the manufacture of a medicament for the treatment and/or alleviation of rheumatoid arthritis, or for the treatment and/or alleviation of inflammation-induced tumors.

10. The application of the compound 20 or pharmaceutically acceptable salt thereof or a pharmaceutical composition or a pharmaceutical preparation containing the compound or the salt thereof in preparing an anti-inflammatory medicament or in preparing a medicament for treating and/or relieving rheumatoid arthritis and/or relieving tumors caused by inflammation;