CN112679492B - Berberine derivative, preparation method and application thereof - Google Patents

Abstract

The invention discloses a berberine derivative, a preparation method and application thereof. The structure of the berberine derivative is shown as a formula I, and the definition of each substituent is described in the specification and the claims. The berberine derivative has anti-tumor activity and can be used for treating diseases such as cancers, lung cancer and the like.

Description

Berberine derivative, preparation method and application thereof

Technical Field

The invention belongs to the field of medicinal chemistry, and relates to a series of derivatives with a natural product berberine structure, a preparation method and application thereof, in particular to the derivatives of the natural product berberine, the preparation method thereof and application thereof as an anti-tumor compound in treating cancers such as lung cancer and the like.

Background

Bererin (BBR) is a fourth class alkaloid of isoquinoline class separated from coptis, has been used as an over-the-counter drug in China for treating diarrhea for decades, and has good safety. BBR has been reported in many studies to have various pharmacological effects, including anti-diarrhea, antibacterial, anti-inflammatory, anti-diabetic, anti-hyperlipidemia, etc. In particular, recent studies have shown that BBR inhibits a variety of tumors, including lymphoma, breast, colon, liver, lung, esophageal, and pancreatic cancers. Many published articles have demonstrated the anti-tumor activity of BBR, revealing that BBR can inhibit the growth of tumor cells, induce apoptosis, inhibit the cell cycle.

It has been reported that BBR can inhibit the migration and invasion of tumor cells by affecting related proteins. For example, E-cadherin (E-cadherin) is an important mediator in the regulation of cell-cell adhesion and is also an important molecule in maintaining epithelial cell morphology and structural integrity. In lung cancer A549 cells, BBR enhances the expression of E-cadherin. It can also inhibit the expression of matrix metalloproteinases, an important class of proteins involved in the degradation of extracellular matrix barriers, involved in the first step of tumor cell metastasis. VEGF (vascular endothelial growth factor) plays a very important role in the neovascularization of tumor cells, while the BBR shows an inhibitory effect on VEGF expression in the melanoma cell line B16F-10. The research also finds that in colon cancer cells, BBR can remarkably inhibit the synthesis of COX-2 and block the activation of a STAT3 signal channel induced by the COX-2, thereby inhibiting the invasion and metastasis of tumor cells. In addition, BBR also affects apoptosis-related proteins and pathways, thereby inhibiting tumor cell proliferation and inducing tumor cell apoptosis. Different tumor cells can activate caspases via the BBR. In addition, in vitro cell experiments report that BBR inhibits the proliferation of various tumor cells by inducing cell cycle arrest.

Studies have shown that mitochondria play a very important role in the anti-tumor role of the BBR. Since most cancer cells have a higher Mitochondrial Membrane Potential (MMP) than normal cells, positively charged alkaloids can selectively accumulate in mitochondria driven by negatively charged Inner Mitochondrial Membrane (IMM). The BBR can reduce mitochondrial membrane potential, inhibit respiratory chain complex I, increase Mitochondrial Permeability Transition (MPT), damage mitochondrial structure, promote mitochondrial oxidative stress and reduce mitochondrial DNA replication numbers, ultimately leading to mitochondrial dysfunction and apoptosis.

The BBR appears to be a relatively safe oral drug and can be used as a drug against several types of malignancies as well as other diseases such as hyperlipidemia and diarrhea. However, the very poor druggability of the BBR results in very low plasma exposure and very low oral bioavailability: (<5%) or acute toxicity by intravenous injection (LD₅₀<10mg/kg) which limits its further development as a potential drug candidate. To overcome these problems, studies have been focused mainly on the modification of C-8, C-9, C-10, C-12 and C-13. Most studies have performed lipophilic modifications by introducing long chain alkyl groups. Such as 8-hexadecyl berberine, can inhibit the growth of lung cancer in vitro and in vivo. Palmitate at position 9 of the BBR shows enhanced lipid lowering efficacy. The 13-octyl berberine derivative has more effective anti-mycobacteria activity. Of course, other non-alkylated derivatives may also improve BBR activity. The 9-O-benzoyl substituted analogues have triglyceride lowering effect. 9-N-substituted BBR derivatives were synthesized and evaluated as a new class of G-tetrad binding ligands or cancer immunotherapeutics. In these studies, the 9-position derivative was a large part of the BBR modification. However, the research on modification of position 9 of BBR to improve its anticancer activity is very limited, and there are few reports.

Disclosure of Invention

The invention aims to provide a berberine derivative which can be used as an anti-cancer drug, so as to open up a new way for discovering drugs for treating cancers such as lung cancer and the like.

The invention also aims to provide a preparation method of the berberine derivative.

The invention also provides the application of the berberine derivative.

In a first aspect of the invention, there is provided a compound of formula I, or an enantiomer, diastereomer, racemate or mixture thereof,

in the formula (I), the compound is shown in the specification,

m is F, Cl, Br or I;

x is O or NH;

y is C (═ O) or CH₂；

R₁、R₂、R₃、R₄Each independently hydrogen, substituted or unsubstituted C₁-C₆Alkyl, substituted or unsubstituted C₃-C₆Cycloalkyl, or substituted or unsubstituted C₆-C₁₂An aryl group;

m is an integer of 1 to 7;

n is an integer of 1 to 7;

R₅and R₆Each independently is hydrogen, hydroxy or carbonyl;

R₇is carboxy, substituted or unsubstituted C₁-C₆Alkyl, substituted or unsubstituted C₁-C₆Alkoxy, substituted or unsubstituted C₁-C₆Alkyloxycarbonyl (-COOC)₁-C₆Alkyl), substituted or unsubstituted C₁-C₆Alkylaminocarbonyl, substituted or unsubstituted C₃-C₁₂Arylaminocarbonyl, substituted or unsubstituted C₆-C₁₂Aryloxycarbonyl, substituted or unsubstituted C₁-C₆Alkylcarbonyl, substituted or unsubstituted C₁-C₆Cycloalkyl carbonyl, or substituted or unsubstituted C₃-C₁₂An arylcarbonyl group;

the above-mentioned substitution means having one or more substituents selected from the group consisting of: c₁-C₆Alkyl radical, C₃-C₁₂Aryl radical, C₃-C₆Cycloalkyl radical, C₁-C₆Alkoxy radical, C₁-C₆Alkylamino, halogen, hydroxy, amino, carboxyl, -COC₁-C₆Alkyl, -COC₃-C₆Cycloalkyl, -COC₆-C₁₂And (4) an aryl group.

In another preferred embodiment, R₅Is hydrogen; r₆Is hydrogen or hydroxy.

In another preferred embodiment, X is O, and Y is C (═ O) or CH₂。

In another preferred embodiment, X is NH and Y is C (═ O).

In another preferred embodiment, R₁、R₂Is hydrogen or C₁-C₄An alkyl group; r₃、R₄Is hydrogen or C₁-C₄An alkyl group.

In another preferred embodiment, R₁、R₂Is hydrogen, R₃、R₄Is hydrogen or C₁-C₄Alkyl, preferably methyl.

In another preferred embodiment, R₁、R₂、R₃、R₄Is C₁-C₄Alkyl, preferably methyl.

In another preferred embodiment, R₇Is carboxyl, C₁-C₄Alkyl, or C₁-C₄An alkyloxycarbonyl group.

In another preferred embodiment, R₇is-COOH, methyl, ethyl, -COOCH₃or-COOCH₂CH₃。

In another preferred embodiment, m is 4,5, 6 or 7; n is 4,5, 6 or 7.

In another preferred embodiment, m is 5 or 6 and n is 5 or 6.

In another preferred embodiment, the compound is:

the compounds of the present invention have asymmetric centers, chiral axes and chiral planes, and can exist in the form of racemates, R-isomers or S-isomers. The person skilled in the art is able to obtain the R-isomer and/or the S-isomer by resolution of the racemate by means of customary technical measures.

In a second aspect of the invention, there is provided a pharmaceutical composition comprising a compound of the first aspect or an enantiomer, diastereomer, racemate or mixture thereof; and

a pharmaceutically acceptable carrier or excipient.

The present invention provides a novel compound which can be used alone or in admixture with pharmaceutically acceptable adjuvants (e.g., excipients, diluents, etc.) to prepare tablets, capsules, granules, syrups, and the like for oral administration. The pharmaceutical composition can be prepared according to a conventional method in pharmacy.

In a third aspect of the present invention, there is provided a use of the compound of the first aspect or the pharmaceutical composition of the second aspect for preparing an antitumor drug.

The present invention also provides a method of treating a tumour comprising the step of administering to a patient in need thereof a compound according to the first aspect or a pharmaceutical composition according to the second aspect.

The present invention also provides a method of inhibiting the growth and/or proliferation of a tumour cell in vitro comprising the step of adding to the tumour cell a compound according to the first aspect or a pharmaceutical composition according to the second aspect.

In another preferred embodiment, the tumor is selected from the group consisting of: lung cancer, lymphoma, breast cancer, colon cancer, liver cancer, esophageal cancer, and pancreas.

The compound of the invention introduces an ETC-1002 derivative long chain at the 9 th position of the berberine, greatly improves the physicochemical properties of the berberine such as cell permeability and drug-forming property, is expected to overcome the defect of poor biological drug-forming property of the berberine, and becomes a possible candidate compound for treating cancers.

It is to be understood that within the scope of the present invention, the above-described features of the present invention and those specifically described below (e.g., in the examples) may be combined with each other to form new or preferred embodiments. Each feature disclosed in this specification may be replaced by an alternative feature serving the same, equivalent, or similar purpose. Not to be reiterated herein, but to the extent of space.

Drawings

Fig. 1 shows the results of the membrane permeability experiment.

Fig. 2 shows the results of the TMRE mitochondrial membrane potential experiments.

Fig. 3 shows the mitochondrial morphology observations.

Fig. 4 shows the results of the effect of compounds on mitochondrial oxygen consumption, where FCCP: uncoupler, R/A is rotenone/antimycin A.

Detailed Description

The inventors of the present application have extensively and intensively studied and tried to introduce a long chain of substituted alkoxy, alkylcarbonyloxy or alkylcarbonylamino substituted ETC-1002 derivatives at the 9-position of berberine, thereby synthesizing a series of berberine derivatives. The introduced substituent has a structure similar to that of the known antihyperlipidemic drug ETC-1002. ETC-1002 is a new drug candidate for the treatment of cardiovascular diseases, and is currently in the third clinical stage. This series of berberine derivatives was evaluated by their effect on survival, antiproliferation and mitochondrial function of a549 lung cancer cell line. Compared with BBR, part of the compounds have better inhibitory activity on A549 lung cancer cell strains. In an anti-proliferation experiment, the derivative has obvious inhibition on the proliferation of an A549 lung cancer cell strain at a lower concentration. Has stronger influence on the mitochondrial function of the A549 cell strain, thereby showing better anticancer effect than berberine. The introduction of ETC-1002 and derivatives greatly improves the physicochemical properties of berberine, such as cell permeability and drug-forming property, is expected to overcome the defect of poor biological drug-forming property of berberine, and becomes a possible candidate compound for treating cancer. On the basis of this, the present invention has been completed.

Term(s) for

In the present invention, the halogen is F, Cl, Br or I.

In the present invention, unless otherwise specified, the terms used have the ordinary meanings well known to those skilled in the art.

In the present invention, the term "C₁-C₆"means having 1, 2, 3, 4,5 or 6 carbon atoms," C₆-C₁₂"means having 6, 7, 8, 9, 10, 11, or 12 carbon atoms, and so forth.

In the present invention, the term "alkyl" denotes a saturated linear or branched hydrocarbon moiety, for example the term "C₁-C₆Alkyl "means a straight or branched chain alkyl group having 1 to 6 carbon atoms, including, but not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, pentyl, hexyl, and the like; preference is given to ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl and tert-butyl.

In the present invention, the term "C₁-C₆Alkoxy "denotes a-O- (C1-6 alkyl) group. For example, the term "C₁-C₆Alkoxy "means a straight or branched chain alkoxy group having 1 to 6 carbon atoms, including, but not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, butoxy, and the like.

In the present invention, the term "cycloalkyl" denotes a saturated cyclic hydrocarbon moiety, for example the term "C₃-C₆Cycloalkyl "refers to cyclic alkyl groups having 3 to 6 carbon atoms in the ring, including without limitation cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like.

In the present invention, the term "aryl" denotes a hydrocarbyl moiety comprising one or more aromatic rings. For example, the term "C₆-C₁₂Aryl "refers to an aromatic ring group having 6 to 12 carbon atoms, such as phenyl, naphthyl, and the like, which does not contain heteroatoms in the ring. The term "C₆-C₁₀Aryl "has a similar meaning. Examples of aryl groups include, but are not limited to, phenyl (Ph), naphthyl, pyrenyl, anthracenyl, and phenanthrenyl.

In the invention, the substitution is mono-substitution or multi-substitution, and the multi-substitution is di-substitution, tri-substitution, tetra-substitution or penta-substitution. By disubstituted is meant having two substituents and so on.

Preparation method

The invention provides a preparation method of berberine derivatives shown in formula I. The preparation method can be realized by the following route.

The preparation of intermediates 1-4 has been reported and can be prepared in a simpler manner.

Specifically, the method comprises the following steps:

(1) commercial or self-made substituted pentadecyl acid 1 generates corresponding acyl chloride under the condition of oxalyl chloride;

(2) condensing the intermediate acyl chloride 6 and berberine to generate different substituted compounds I-I;

(3) condensing the intermediate acyl chloride 6 and the intermediate 4 to generate amide compounds I-II;

(4) condensing halide of pentadecyl with different substituents and berrubine to generate ether compounds I-III;

wherein the content of the first and second substances,

the solvent used for the reaction with oxalyl chloride in the step (1) is selected from solvents such as dichloromethane and the like; the reaction temperature is 0 ℃;

the solvent used in the condensation reaction in the step (2) is selected from acetonitrile; the reaction temperature is 90 ℃;

the solvent used in the condensation reaction in the step (3) is selected from acetonitrile; the reaction temperature is 90 ℃;

the solvent used in the condensation reaction in the step (4) is selected from acetonitrile; the reaction temperature is 90 ℃;

in the step (4), the halide can be a fluoride, a chloride, a bromide and an iodide;

pharmaceutical composition

The invention also provides a pharmaceutical composition comprising a safe and effective amount of the active ingredient, and a pharmaceutically acceptable carrier.

The active ingredient refers to the compound of the formula I.

The active ingredient and the pharmaceutical composition are used for preparing the drugs for treating tumors.

"safe and effective amount" means: the amount of active ingredient is sufficient to significantly improve the condition without causing serious side effects. Typically, the pharmaceutical composition contains 1-2000mg of active ingredient per dose, more preferably, 10-200mg of active ingredient per dose. Preferably, said "dose" is a tablet.

"pharmaceutically acceptable carrier" refers to: one or more compatible solid or liquid fillers or gel substances which are suitable for human use and must be of sufficient purity and sufficiently low toxicity. By "compatible" is meant herein that the components of the composition are capable of being combined with the active ingredients of the present invention and with each other without significantly diminishing the efficacy of the active ingredient. Examples of pharmaceutically acceptable carrier moieties are cellulose and its derivatives (e.g., sodium carboxymethylcellulose, sodium ethylcellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (e.g., stearic acid, magnesium stearate), calcium sulfate, vegetable oils (e.g., soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (e.g., propylene glycol, glycerin, mannitol, sorbitol, etc.), emulsifiers (e.g., propylene glycol, glycerin, mannitol, sorbitol, etc.), and the like

) Wetting agents (e.g., sodium lauryl sulfate), coloring agents, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, and the like.

The mode of administration of the active ingredient or pharmaceutical composition of the present invention is not particularly limited, and representative modes of administration include (but are not limited to): oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous), and the like.

Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active ingredient, the liquid dosage forms may contain inert diluents commonly employed in the art such as, for example, water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, propylene glycol, 1, 3-butylene glycol, dimethylformamide and oils, especially cottonseed, groundnut, corn germ, olive, castor and sesame oils or mixtures of such materials and the like. In addition to these inert diluents, the compositions can also contain adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

Suspensions, in addition to the active ingredients, may contain suspending agents as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these materials, and the like.

Compositions for parenteral injection may comprise physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and nonaqueous carriers, diluents, solvents or vehicles include water, ethanol, polyols and suitable mixtures thereof.

The compounds of the present invention may be administered alone or in combination with other therapeutic agents, such as chemotherapeutic agents.

When the pharmaceutical composition is used, a safe and effective amount of the compound of the present invention is suitable for mammals (such as human beings) to be treated, wherein the administration dose is a pharmaceutically-considered effective administration dose, and for a human body with a weight of 60kg, the daily administration dose is usually 1 to 2000mg, preferably 20 to 500 mg. Of course, the particular dosage will depend upon such factors as the route of administration, the health of the patient, and the like, and is within the skill of the skilled practitioner.

The invention will be further illustrated with reference to the following 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 invention. The experimental procedures for which specific conditions are not indicated in the following examples are generally carried out according to conventional conditions (e.g.as described in Sambrook et al, molecular cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989)) or according to the conditions as recommended by the manufacturer. Unless otherwise indicated, percentages and parts are percentages and parts by weight.

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. In addition, any methods and materials similar or equivalent to those described herein can be used in the methods of the present invention. The preferred embodiments and materials described herein are intended to be exemplary only.

In the following examples, NMR was measured using a Mercury-Vx 300M instrument manufactured by Varian, NMR calibration: Δ H7.26 ppm (CDCl)₃)，2.50ppm(DMSO-d₆)，3.15ppm(CD₃OD); reagents are mainly provided by Shanghai chemical reagents company; TLC thin layer chromatography silica gel plate is produced by Shandong tobacco Taihuyou silica gel development Co., Ltd, model number HSGF 254; the normal phase column chromatography silica gel used for compound purification is produced by Shandong Qingdao ocean chemical plant, model zcx-11, 200-300 mesh.

Example 1

Synthesis of intermediate 5: the starting material ETC-1002(1eq,0.96mmol) was dissolved in dichloromethane, Dess-Martin oxidant (2eq,1.92mmol) was added at 0 deg.C, warmed to room temperature and stirred for 1 hour, and TLC monitored for completion of the reaction. Quenched with saturated sodium thiosulfate solution, extracted with dichloromethane, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography (DCM: MeOH ═ 25:1) to afford the product, intermediate 5(0.82mmol, 86%):¹H NMR(300MHz,CDCl₃)δ2.36(t,J＝6.0Hz,4H),1.61-1.44(m,8H),1.27-1.23(m,8H),1.16(s,12H).ESI-MS(m/z):[M+Na]⁺,365.2.

synthesis of intermediate 6: oxalyl chloride (2.46mmol,3eq) was added dropwise to a solution of intermediate 5(0.82mmol,1eq) in anhydrous dichloromethane at 0 ℃ followed by a catalytic amount of DMF. The mixed solution is warmed to room temperature and stirred for 2h, and the reaction is monitored by TLCCompletely, concentrating under reduced pressure. The crude product was dissolved in anhydrous acetonitrile, added dropwise to a solution of intermediate 1(2.46mmol,3eq) in anhydrous acetonitrile, warmed to reflux and stirred overnight. Quench it with 1mL methanol, concentrate the mixture under reduced pressure and purify by column chromatography (DCM: MeOH ═ 10:1) to give the product, intermediate 6(0.28mmol, 34%):¹H NMR(300MHz,CDCl₃)δ9.96(s,1H),8.58(s,1H),8.21(d,J＝9.0Hz,1H),7.76(d,J＝9.0Hz,1H),7.42(s,1H),6.75(s,1H),6.03(s,2H),5.29(s,2H),3.95(s,3H),3.64(s,3H),3.25(s,2H),2.45-2.35(m,4H),2.02-1.54(m,8H),1.32-1.14(m,20H).ESI-MS(m/z):[M-Cl]⁺,660.4.

synthesis of intermediate 7: intermediate 6(0.28mmol,1eq) was dissolved in methanol at 0 ℃, sodium borohydride (2.8mmol,10eq) was added carefully, stirred for 30min at 0 ℃ and monitored by TLC for completion. Concentration under reduced pressure removed methanol, dilution with ethyl acetate, water wash, brine wash, drying over anhydrous sodium sulfate, concentration under reduced pressure, and column chromatography purification (DCM: MeOH ═ 50:1) afforded intermediate 7(0.22mmol, 80%):¹H NMR(300MHz,CDCl₃)δ6.98(d,J＝6.0Hz,1H),6.81(d,J＝9.0Hz,1H),6.72(s,1H),6.58(s,1H),5.91(s,2H),3.98(d,J＝15.0Hz,1H),3.77(s,3H),3.65(s,3H),3.59-3.55(m,2H),3.44(d,J＝15.0Hz,1H),3.24-3.17(m,1H),3.12-3.05(m,2H),2.86-2.77(m,1H),2.67-2.60(m,2H),1.80-1.11(m,32H).ESI-MS(m/z):[M+H]⁺,666.5.

synthesis of compound B1: potassium acetate (0.48mmol,2.2eq) and intermediate 7 were dissolved in ethanol, iodine (0.44mmol,2eq) was added, and the mixture was stirred at room temperature for 2h, protected from light. After dilution with dichloromethane, the organic phase was washed with sodium thiosulfate solution, saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the crude product was acidified with 1N diluted hydrochloric acid and purified by column chromatography (DCM: MeOH ═ 10:1) to give compound B1(0.11mmol, 50%):¹H NMR(500MHz,CD₃OD)δ9.31(s,1H),8.83(s,1H),8.19(dd,J＝14.8,9.2Hz,2H),7.69(s,1H),6.97(s,1H),6.12(s,2H),4.94(t,J＝6.0Hz,2H),4.08(s,3H),3.64(s,3H),3.53(s,1H),3.26(t,J＝6.4Hz,2H),1.87-1.83(m,2H),1.53-1.40(m,18H),1.30-1.14(m,12H).¹³C NMR(125MHz,CD₃OD)δ180.1,176.5,152.4,150.0,144.7,140.2,135.9,135.2,132.0,127.9,127.0,123.1,122.1,121.7,109.4,106.7,103.7,72.3,57.9,57.7,52.2,44.4,43.4,41.9,41.8,38.4,31.5,31.2,28.1,26.8,26.6,26.1,25.7,25.6.HRMS(TOF ESI)calcd for C₃₉H₅₂ClNO₈[M-Cl]⁺:662.3687；found:662.3693.

the following compounds were synthesized in the same manner:

compounds B2, B8 and B9 were prepared from intermediate 4 by a similar synthesis method as in example 1.

Compound B2

¹H NMR(500MHz,CD3OD)δ9.32(s,1H),8.84(s,1H),8.23(q,J＝9.0Hz,2H),7.70(s,1H),6.98(s,1H),6.12(s,2H),4.94(t,J＝6.0Hz,2H),4.08(s,3H),3.53(s,1H),3.27(t,J＝6.5Hz,2H),1.87-1.43(m,20H),1.29-1.14(m,12H).¹³C NMR(126MHz,MeOD)δ176.5,152.4,150.0,144.6,140.3,135.9,135.3,132.1,127.9,127.0,123.1,122.2,121.7,109.4,106.7,103.8,72.4,57.8,57.7,44.4,43.1,42.0,41.8,38.4,31.4,28.1,26.8,26.7,26.2,26.1,25.8,25.6.

Compound B8

¹H NMR(600MHz,CD₃OD)δ9.19(s,1H),8.79(s,1H),8.27(d,J＝9.6Hz,1H),8.16(d,J＝9.6Hz,1H),7.68(s,1H),6.97(s,1H),6.11(s,2H),4.94(t,J＝6.0Hz,2H),4.09(s,3H),3.64(s,3H),3.52(s,1H),3.27(t,J＝6.0Hz,2H),1.79-1.76(m,2H),1.53-1.38(m,17H),1.34-1.20(m,7H),1.16-1.15(m,6H).¹³C NMR(151MHz,MeOD)δ181.5,180.1,156.6,152.3,150.0,146.6,139.7,135.4,131.9,129.5,126.0,122.8,122.2,121.7,109.4,106.6,103.7,72.3,57.8,57.6,52.2,44.4,43.4,42.5,41.9,38.4,31.5,31.3,28.2,26.9,26.7,26.2,26.1,25.9,25.6,25.6.

Compound B9

¹H NMR(500MHz,CD3OD)δ9.17(s,1H),8.80(s,1H),8.27(d,J＝9.5Hz,1H),8.17(d,J＝9.0Hz,1H),7.69(s,1H),6.97(s,1H),6.12(s,2H),4.93(t,J＝6.5Hz,2H),4.10(s,3H),3.52(s,1H),3.27(t,J＝6.0Hz,2H),1.79-1.75(m,2H),1.52-1.25(m,24H),1.14(s,6H).¹³C NMR(126MHz,MeOD)δ181.5,156.6,152.3,150.0,146.5,139.7,135.5,131.9,129.5,126.1,122.8,122.3,121.7,109.4,106.6,103.7,72.4,57.8,57.6,44.4,43.1,42.5,42.0,38.4,31.5,31.4,28.2,26.9,26.7,26.2,25.9,25.8.

EXAMPLE 2 Synthesis of Compound B3

Reagent 13(0.83mmol,1eq) was dissolved in anhydrous dichloromethane, oxalyl chloride (1.65mmol,2eq) and catalytic amount of DMF were added dropwise at 0 deg.C, warmed to room temperature and stirred for 2 h. The reaction solution was concentrated under reduced pressure, and the crude anhydrous acetonitrile solution was added dropwise to the anhydrous acetonitrile solution of intermediate 1(0.83mmol,1eq), and the mixture was heated to reflux and stirred overnight. The reaction was concentrated under reduced pressure and the crude product was purified by column chromatography (DCM: MeOH ═ 15:1) to afford compound B3(0.50mmol, 60%):¹H NMR(400MHz,CD3OD)δ9.71(s,1H),8.82(s,1H),8.22(dd,J＝13.6,9.6Hz,2H),7.69(s,1H),6.97(s,1H),6.12(s,2H),4.94(t,J＝6.4Hz,2H),4.08(s,3H),3.27(d,J＝6.4Hz,2H),2.86(t,J＝7.2Hz,2H),1.88-1.80(m,2H),1.53(s,2H),1.33-1.30(m,20H),0.90(t,J＝6.0Hz,3H).¹³C NMR(100MHz,CD3OD)δ172.4,152.4(2C),150.0,145.3,140.2,135.8,135.1,132.0,127.9,127.0,123.3,122.0,121.7,109.4,106.6,103.7,57.7,57.3,34.5,33.1,30.8(2C),30.7(2C),30.5(2C),30.1,28.1,25.9,23.7,14.4.HRMS(TOF ESI)calcd for C₃₄H₄₄ClNO₅[M-Cl]⁺:546.3214；found:546.3214.

the following compounds were synthesized in the same manner:

compound B4

¹H NMR(400MHz,CD₃OD)δ9.72(s,1H),8.82(s,1H),8.22(dd,J＝15.2,9.6Hz,2H),7.69(s,1H),6.97(s,1H),6.12(s,2H),4.95(t,J＝6.4Hz,2H),4.08(s,3H),3.26(t,J＝6.4Hz,2H),2.87(t,J＝7.6Hz,2H),2.26(t,J＝7.6Hz,2H),1.88-1.80(m,2H),1.59-1.53(m,4H),1.32(s,16H).¹³C NMR(100MHz,DMSO)δ183.9,180.1,159.8,159.5,157.2,153.8,147.6,143.0,142.4,140.3,136.1,135.4,130.6,130.0,129.8,117.9,115.0,111.6,66.7,64.8,43.1,42.6,38.4,38.4(2C),38.2,38.0,37.8,35.6,34.0,33.7.

Compound B5

¹H NMR(400MHz,CD₃OD)δ9.72(s,1H),8.81(s,1H),8.21(dd,J＝14.4,9.2Hz,2H),7.69(s,1H),6.97(s,1H),6.12(s,2H),4.95(t,J＝6.0Hz,2H),4.08(s,3H),4.04(t,J＝6.8Hz,2H),3.26(t,J＝6.0Hz,2H),2.87(t,J＝7.2Hz,2H),2.01(s,3H),1.88-1.80(m,2H),1.65-1.50(m,4H),1.44-1.32(m,18H).¹³C NMR(100MHz,CD₃OD_SPE)δ172.9,172.2,152.3,152.2,149.8,145.1,140.1,135.6,134.9,131.9,127.8,126.8,123.1,121.8,121.6,109.2,106.5,103.6,65.5,57.6,57.1,34.4,30.5,30.5,30.3,30.2,29.9,29.5,27.9,26.8,25.7,20.6,9.0.

Compound B6

¹H NMR(400MHz,CD₃OD)δ9.71(s,1H),8.81(s,1H),8.21(dd,J＝13.2,8.8Hz,2H),7.69(s,1H),6.97(s,1H),6.12(s,2H),4.94(t,J＝5.6Hz,2H),4.08(s,3H),3.64(s,3H),3.26(t,J＝6.0Hz,2H),2.86(t,J＝7.2Hz,2H),2.31(t,J＝7.2Hz,2H),1.88-1.80(m,2H),1.61-1.50(m,4H),1.32-1.30(m,16H).¹³C NMR(100MHz,DMSO)δ182.8,180.1,159.8,159.4,157.2,153.9,147.6,143.1,142.4,140.3,136.1,135.4,130.6,130.1,129.8,117.9,115.0,111.6,66.7,64.7,60.6,43.1,42.7,38.4,38.3,38.2,38.1,37.9,37.8,35.6,33.9,33.7.

Compound B7

¹H NMR(400MHz,CD₃OD)δ9.73(s,1H),8.81(s,1H),8.20(dd,J＝16.8,9.2Hz,2H),7.68(s,1H),6.97(s,1H),6.11(s,2H),4.96(t,J＝6.0Hz,2H),4.07(s,3H),3.54(s,1H),3.26(t,J＝6.0Hz,2H),2.88(t,J＝7.2Hz,2H),1.89-1.81(m,2H),1.58-1.53(m,3H),1.49-1.42(m,7H),1.31(s,10H),0.90(t,J＝6.8Hz,3H).¹³C NMR(100MHz,CD₃OD_SPE)δ172.4,152.5,152.4,150.0,145.3,140.3,135.8,135.1,132.1,128.0,127.0,123.3,122.0,121.8,109.4,106.7,103.8,72.5,57.8,57.4,38.6,38.4,34.6,33.1,30.9,30.6,30.5,30.2,28.1,26.9,26.8,25.9,23.8,14.5.

Example 3

Synthesis of intermediate 25:1, 7-dibromoheptane (0.26mmol,5eq) and potassium tert-butoxide (0.075mmol,1.5eq) were dissolved in anhydrous DMF, and a solution of the starting material 24(0.05mmol,1eq) in anhydrous DMF at 0 ℃ was added dropwise to the solution, and the mixture was stirred at room temperature for 2 h. The reaction mixture was diluted with ethyl acetate, washed with water and saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was dissolved in DCM, 0.1mL of concentrated HCl was added and stirred vigorously for 2 h. Quenching with saturated sodium carbonate solution, extracting with ethyl acetate, washing with saturated sodium chloride solution, and drying with anhydrous sodium sulfate. Column chromatography purification (PE: EA ═ 10:1) afforded intermediate 25(0.02mmol, 39%):¹H NMR(400MHz,CDCl₃)δ4.11(q,J＝8.0Hz,2H),3.40(t,J＝4.0Hz,2H),2.40-2.35(m,4H),1.88-1.81(m,2H),1.58-1.54(m,5H),1.51-1.47(m,2H),1.44-1.38(m,2H),1.31-1.29(m,3H),1.24(t,J＝8.0Hz,7H),1.14(s,6H).ESI-MS(m/z):[M+H]⁺,391.2.

synthesis of intermediate 26: intermediate 25 was dissolved in methanol, sodium borohydride (0.2mmol,10eq) was carefully added at 0 ℃ and stirred for 30min before concentration under reduced pressure, the crude product was diluted with DCM, washed with water and dried over anhydrous sodium sulfate, concentrated under reduced pressure and purified by column chromatography (PE: EA ═ 10:1) to afford intermediate 26(0.019mmol, 99%):¹H NMR(400MHz,CDCl₃)δ4.11(q,J＝8.0Hz,2H),3.57(s,1H),3.40(t,J＝4.0Hz,2H),1.89-1.81(m,2H),1.60(s,1H),1.52-1.48(m,2H),1.42-1.38(m,7H),1.35-1.27(m,7H),1.24(t,J＝8.0Hz,6H),1.15(s,6H).ESIMS(m/z):[M+H]⁺,393.4.

synthesis of compound B10: intermediate 26(0.03mmol,1eq) and intermediate 1(0.092mmol,3eq) were dissolved in anhydrous acetonitrile, warmed to reflux and stirred overnight. Concentrating the reaction solution under reduced pressure, adding 1N HCl for acidification, and purifying by column chromatography (DCM)MeOH ═ 25:1) gave compound B10(0.018mmol, 60%):¹H NMR(400MHz,CD₃OD)δ9.68(s,1H),8.71(s,1H),8.12(d,J＝9.2Hz,1H),8.00(d,J＝9.2Hz,1H),7.67(s,1H),6.97(s,1H),6.11(s,2H),4.94(t,J＝6.0Hz,2H),4.41(t,J＝6.8Hz,2H),4.12-4.07(m,5H),3.51(s,1H),3.26(t,J＝6.4Hz,2H),1.98-1.91(m,2H),1.57-1.22(m,23H),1.14(s,6H).¹³C NMR(100MHz,CD₃OD)δ179.7,152.2,149.9,146.3,145.1,139.7,135.2,131.9,128.0,124.3,123.6,121.6,109.4,106.6,103.7,75.8,72.3,61.5,57.6,57.3,43.3,41.9,38.4,31.3,31.2,30.8,30.5,28.2,26.9,26.8,26.6,26.1,25.6,14.6.HRMS(TOF ESI)calcd for C₃₈H₅₂ClNO₇[M-Cl]⁺:634.3738；found:634.3742.

synthesis of compound B11: compound B10(0.032mmol,1eq) was dissolved in a mixed solvent of ethanol and water (2:1), sodium hydroxide (0.16mmol,5eq) was added, and the mixture was stirred under reflux for 2 h. The reaction was concentrated under reduced pressure, the crude product was diluted with water and acidified with 1N HCl solution, extracted with DCM, dried over anhydrous sodium sulfate, concentrated under reduced pressure and purified by column chromatography (DCM: MeOH ═ 10:1) to give compound B11(0.016mmol, 50%):¹H NMR(400MHz,CD₃OD)δ9.67(s,1H),8.71(s,1H),8.11(d,J＝9.2Hz,1H),7.99(d,J＝9.2Hz,1H),7.66(s,1H),6.96(s,1H),6.10(s,2H),4.93(t,J＝6.4Hz,2H),4.40(t,J＝6.8Hz,2H),4.09(s,3H),3.50(s,1H),3.25(t,J＝6.4Hz,2H),1.97-1.90(m,2H),1.59-1.28(m,20H),1.13(s,6H).¹³C NMR(100MHz,CD₃OD)δ152.2,152.1,149.9,146.3,145.1,139.7,135.2,131.9,128.0,124.3,123.6,121.9,121.6,109.4,106.6,103.7,75.8,72.4,57.6,57.3,43.1,41.9,38.4,31.4,31.2,30.8,30.5,28.2,26.9,26.8,26.7,26.2,25.7.HRMS(TOF ESI)calcd for C₃₆H₄₈ClNO₇[M-Cl]⁺:606.3425；found:606.3425.

the following compounds were synthesized in the same manner:

compound B12

¹H NMR(400MHz,CD3OD_SPE)δ9.66(s,1H),8.67(s,1H),8.09-7.97(m,2H),7.63(s,1H),6.93(s,1H),6.11(s,2H),4.93(t,J＝6.4Hz,2H),4.41(t,J＝6.8Hz,2H),4.10(s,3H),3.25(t,J＝6.4Hz,2H),1.96-1.89(m,2H),1.58-1.51(m,2H),1.43-1.40(m,2H),1.27(s,20H),0.88(t,J＝6.8Hz,3H).

¹³C NMR(100MHz,CD₃OD_SPE)δ152.1,151.9,149.9,146.0,145.0,139.5,135.1,131.6,128.0,124.2,123.5,121.7,121.6,109.4,106.5,103.6,75.8,57.6,57.2,33.0,31.1,30.7,30.5,30.4,28.2,26.9,23.7,14.5.

Compound B13

¹H NMR(400MHz,CD3OD_SPE)δ9.68(s,1H),8.71(s,1H),8.13-7.99(m,2H),7.67(s,1H),6.97(s,1H),6.11(s,2H),4.94(t,J＝6.4Hz,2H),4.41(t,J＝6.8Hz,2H),4.10(s,3H),3.65(s,3H),3.26(t,J＝6.0Hz,2H),2.31(t,J＝7.6Hz,2H),1.97-1.90(m,2H),1.59-1.52(m,4H),1.45-1.30(m,18H).

¹³C NMR(101MHz,CD3OD_SPE)δ176.0,152.2,152.1,149.9,146.3,145.1,139.7,135.2,131.9,128.0,124.3,123.6,121.9,121.6,109.4,106.6,103.7,75.8,57.6,57.3,52.0,34.8,31.2,30.7(3C),30.6,30.5,30.4,30.2,28.2,27.0,26.0.

Compound B14

¹H NMR(400MHz,CD3OD_SPE)δ9.68(s,1H),8.71(s,1H),8.06(dd,J＝48.8,9.2Hz,2H),7.67(s,1H),6.97(s,1H),6.11(s,2H),4.94(t,J＝6.0Hz,2H),4.41(t,J＝6.8Hz,2H),4.10(s,3H),3.26(t,J＝6.0Hz,2H),2.24(t,J＝7.2Hz,2H),1.97-1.90(m,2H),1.62-1.52(m,4H),1.46-1.41(m,2H),1.30(s,16H).

¹³C NMR(100MHz,CD3OD_SPE)δ152.2,152.1,150.0,146.3,145.1,139.7,135.2,131.9,128.0,124.3,123.6,121.9,121.6,109.4,106.6,103.7,75.8,57.6,57.3,35.9,31.2,30.7(2C),30.6,30.5(2C),30.5,30.4,28.2,27.0,26.5.

Compound B15

¹H NMR(400MHz,CD3OD_SPE)δ9.67(s,1H),8.70(s,1H),8.05(dd,J＝44.4,9.2Hz,2H),7.65(s,1H),6.96(s,1H),6.10(s,2H),4.95(t,J＝6.0Hz,2H),4.40(t,J＝6.4Hz,2H),4.12-4.07(m,5H),3.26(t,J＝6.0Hz,2H),1.97-1.90(m,2H),1.57-1.48(m,4H),1.45-1.22(m,21H),1.14(s,6H).

¹³C NMR(100MHz,CD3OD_SPE)δ179.7,152.2,152.1,149.9,146.3,145.1,139.6,135.2,131.9,128.0,124.3,123.6,121.9,121.6,109.4,106.6,103.7,75.9,61.5,57.6,57.3,43.3,41.9,31.2,31.1,30.7,30.6,30.5,28.2,27.0,26.0,25.6,14.6.

Example 4

Test example for inhibition of A549 cell line

1. Purpose of experiment

The series of compounds are tested for anti-tumor activity, and the in vitro anti-tumor activity of the compounds is evaluated by measuring the growth inhibition activity of the compounds on human tumor cells.

2. Principle of experiment

The analysis method uses tetrazole compound [3- (4, 5-dimethylthiozol-2-yl) -5- (3-carboxymethyloxy phenyl) -2- (4-sulfophenyl) -2H-tetrazolium, inner salt; MTS ] as the basis. The conversion of yellow MTS into blue-violet water-soluble formazan is catalyzed by dehydrogenase, and the dehydrogenase exists in metabolically active living cells; dead cells cannot complete the conversion of MTS to formazan. The amount of Formazan (Formazan) determined by absorbance at 490nm is directly proportional to the number of viable cells in culture.

3. Procedure of experiment

Will be 3X 10³Individual a549 cells were seeded in triplicate in each well of 96 wells and incubated with increasing concentrations of different compounds for 72 hours. Add 20. mu.L of CellTiter96 AQueous One Solution reagent (Promega) per well. Absorbance at 490nm was measured using a Spectramax microplated reader 340PC 384. Will IC₅₀Values are reported as the average of three independent experiments performed in triplicate. IC (integrated circuit)₅₀The inhibitor concentration was measured in the range of 0 to 200. mu. mol/L.

4. Results of the experiments (the following compounds are given as examples and are not limiting)

TABLE 1 inhibitory Effect of Compounds on A549 cell line

The results show that the compounds have the inhibitory activity on A549 cell strains, and the inhibitory activity of partial compounds is obviously improved compared with berberine.

Example 5

Permeability study of Compounds on A549 cell line

1. Purpose of experiment

The permeability of the compound was evaluated by measuring the concentration of the compound in a549 cell line.

2. Experimental procedure

Mixing 2.5X 10⁵A549 cells were seeded in 12-well plates in triplicate and incubated overnight at 37 ℃. After 8 hours incubation with 5 μ M of the different compounds, the medium was aspirated from the wells. Cells were washed three times with ice PBS and lysed in 1mL lysis buffer (acetonitrile: PBS ═ 1: 2) on ice for 20 minutes. After sonication, the collected supernatant was centrifuged at 12000rpm (4 ℃) for 10 minutes to remove cell debris. The supernatant was dried under vacuum, then 100. mu.L of acetonitrile was added to the residue, respectively, and after vortexing the mixture for 3 minutes and centrifuging at 10000rpm for 5 minutes, the supernatant was collected and the concentration of the compound was determined by LC-MS.

3. Results of the experiment

The results of the membrane permeability experiments for some of the compounds are shown in FIG. 1. The results show that the concentration of B1 and B10 in cells is greatly increased compared with berberine, which provides possibility for improving the pharmacokinetic property of the compound and further improves the pharmaceutical property.

Example 6

Effect on mitochondrial function

1. Influence on the Membrane potential

1.1 purpose of the experiment

The change of the A549 mitochondrial membrane potential is measured by using a fluorescence labeling method, and the influence of the berberine derivative on the mitochondrial membrane potential at the level of intact cells is detected, so that the compound with improved anticancer activity is obtained.

1.2 principle of the experiment

TMRE is a positively charged dye rhodamine that selectively localizes to mitochondria and is thus very troublesome for labeling mitochondria in living cells. Besides selectively dyeing mitochondria, the method can be widely used for setting a smear.

1.3 Experimental procedures

A549 cells were seeded at a density of 300000cells/mL in 96-well plates and incubated overnight and supplemented with 10% FBS F12 medium. Cells were then treated with different concentrations of the compounds for 24 hours. 100nM TMRE and 1. mu.g/ml Hoechest 33342 were added to each well and incubated in the dark 30min and 10 min before the end of the treatment, respectively. Cells were analyzed by an Operetta high content screening system. Images of TMRE and Hoechst were observed under a 40x microscope, and the fluorescence intensity of TMRE was measured.

1.4 results of the experiment (the following compounds are given as examples and are not limiting)

The results of the TMRE mitochondrial membrane potential experiments are shown in FIG. 2. The results show that compounds B1 and B10 have an effect on mitochondrial membrane potential, with B1 having a weaker effect on membrane potential at a concentration of 10 μ M, and compound B10 having an effective, significant and dose-dependent effect on membrane potential. While berberine had no effect on mitochondria at a concentration of 10 μ M.

2. Effect on mitochondrial architecture

2.1 purpose of the experiment

The improvement of the anticancer activity is further explained by observing the structure change of mitochondria and detecting the influence of the berberine derivative on the mitochondria.

2.2 principle of the experiment

Mitochondrial morphology was observed by a Mito-Tracker green fluorescent marker.

2.3 Experimental procedures:

a549 cells were seeded at a density of 150000cells/mL in 96-well plates and incubated overnight and supplemented with 10% FBS F12 medium. Cells were then treated with different concentrations of the compounds for 24 hours. At 20 and 10 minutes before the end of incubation, 100nM Mito Tracker Green and a final concentration of 1. mu.g/ml Hoechest 33342 were added to each well and incubation continued in the dark. After replacing the medium with PBS, the cells were analyzed by a Phenix high content screening system. Confocal images of Mito Tracker Green and Hoechst were taken under a 63x microscope.

2.4 Experimental results: (the following compounds are exemplified but not limited to)

The results of mitochondrial morphology observation with Mito-Tracker green fluorescent label are shown in FIG. 3.

The results show that the compound B1 has slight change of the mitochondrial reticular structure at the concentration of 10 mu M, while the compound B10 has obvious fragmentation of the mitochondrial structure at the concentration of 10 mu M, and berberine has no obvious influence on the mitochondria at the concentration.

3. Effect on mitochondrial Oxygen Consumption (OCR)

3.1 purpose of the experiment

The influence of the derivatives on OCR of a549 cells was explored.

3.2 principle of the experiment

The change of the aerobic respiration capacity of the mitochondria of the cells is reflected by detecting the oxygen consumption rate in each micro-chamber.

3.3 Experimental procedures

A549 cells were plated at 30w/ml in 96-well plates at 80. mu.l per well. Adherent growth was 24 h. The medium was then replaced with pre-heated Oxygen Consumption Rate (OCR) assay buffer after gently rinsing the cells with PBS. The final concentrations of compound for the four injections in the measurement were as follows: injection of a, various concentrations of test compound; injection of B, oligomycin 1. mu.M as ATP synthase inhibitor; injection of C, FCCP 0.6 μ M as mitochondrial uncoupling agent; injection D, rotenone and antimycin 10. mu.M each as an inhibitor of oxidative phosphorylation of the electron transport chain. In the absence of CO₂After incubation at 37 ℃ for about 1 hour, OCR of the cells was measured.

3.4 results of the experiment (the following compounds are given as examples and are not limiting)

The results of the effect of the compounds on mitochondrial oxygen consumption are shown in figure 4.

The results show that both compounds B1 and B10 significantly and dose-dependently reduced mitochondrial oxygen consumption.

All documents referred to herein are incorporated by reference into this application as if each were individually incorporated by reference. Furthermore, it should be understood that various changes and modifications of the present invention can be made by those skilled in the art after reading the above teachings of the present invention, and these equivalents also fall within the scope of the present invention as defined by the appended claims.

Claims (15)

1. A compound of the formula I, wherein,

in the formula (I), the compound is shown in the specification,

m is F, Cl, Br or I;

x is O, Y is C (═ O) or CH₂(ii) a Or X is NH and Y is C (═ O);

R₁、R₂、R₃、R₄each independently hydrogen or substituted or unsubstituted C₁-C₆An alkyl group;

m is an integer of 1 to 7;

n is an integer of 1 to 7;

R₅and R₆Each independently is hydrogen or hydroxy;

R₇is carboxy, substituted or unsubstituted-COOC₁-C₆An alkyl group;

the above-mentioned substitution means having one or more substituents selected from the group consisting of: c₁-C₆An alkyl group.

2. The compound of claim 1, wherein R is₅Is hydrogen; r₆Is hydrogen or hydroxy.

3. A compound according to claim 1, wherein X is O, and Y is C (═ O) or CH₂。

4. As claimed in claim 1The compound of (1), wherein R is₁、R₂Is hydrogen or C₁-C₄An alkyl group; r₃、R₄Is hydrogen or C₁-C₄An alkyl group.

5. The compound of claim 1, wherein R is₁、R₂Is hydrogen, R₃、R₄Is hydrogen or C₁-C₄An alkyl group.

6. The compound of claim 1, wherein R is₁、R₂、R₃、R₄Is C₁-C₄An alkyl group.

7. The compound of claim 1, wherein R is₇Is carboxyl or C₁-C₄An alkyloxycarbonyl group.

8. The compound of claim 1, wherein R is₇is-COOH, -COOCH₃or-COOCH₂CH₃。

9. The compound of claim 1, wherein m is 4,5, 6, or 7; n is 4,5, 6 or 7.

10. The compound of claim 1, wherein m is 5 or 6 and n is 5 or 6.

11. A compound of the formula I, wherein,

the compound is:

12. a pharmaceutical composition, comprising:

a compound according to any one of claims 1 to 11; and

a pharmaceutically acceptable carrier.

13. Use of a compound according to any one of claims 1 to 11 or a pharmaceutical composition according to claim 12 for the preparation of an anti-tumor medicament.

14. The use of claim 13, wherein the tumor is selected from the group consisting of: lung cancer, lymphoma, breast cancer, colon cancer, liver cancer, esophageal cancer, and pancreas.

15. A method for non-therapeutically inhibiting the growth and/or proliferation of tumor cells in vitro comprising the step of adding to the tumor cells a compound according to any one of claims 1 to 11 or a pharmaceutical composition according to claim 12.

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