KR20100062072A - Novel coumarin based compound or pharmaceutically acceptable salt thereof, preparation method thereof and pharmaceutical composition for inhibition of multidrug resistance containing the same as an active ingredient - Google Patents

Abstract

PURPOSE: A pharmaceutical compositioin containing a novel coumarin compounds for suppressing multidrug resistance is provided to suppress overexpression of P-glycoprotein and to enhance anticancer activity without side effects. CONSTITUTION: A coumarin compounds is denoted by chemical formula 1. In chemical formula 1, X is alkylene of C1-C6 or carbonyl and Y is phenyl or pyridyl. The coumarin compounds of chemical formula 1 is courmarin compounds of 6-(3-methyl-but-2-enyl)-7-(4-nitro-benzyloxy)-chromen-2-one or 6-(3-methyl-but-2-enyl)-7-(pyridine-2-ylmethoxy)-chromen-2-one. A coumarin compound of chemical formula 1(a) is prepared by reacting demethylsuberosin of chemical formula 7 with acyl halide under the presence of base through esterificaiton. A pharmaceutical composition for suppressing multidrug resistance containts the coumarin compounds of chemical formula 1 as an active ingredient.

Description

Novel coumarin based compound or pharmaceutical acceptable salts, preparation method, etc. and pharmaceutical composition for inhibition of multidrug resistance containing the same as an active ingredient}

The present invention relates to a coumarin compound, a preparation method thereof and a pharmaceutical composition for inhibiting multi-drug resistance containing the same as an active ingredient.

Until now, various treatment methods such as surgical therapy, radiation therapy, and chemotherapy for the treatment of cancer have been studied, but the situation has not been reached until the full treatment of cancer.

Surgical and radiation therapy is a useful method in the early stages of cancer development, so the dependence on chemotherapy is gradually increasing in cancers that are difficult to detect early, and the chemotherapy has a history of more than 50 years. Hundreds of anticancer drugs have been developed and used in the clinic until now, but there are not many cases of obtaining clinically satisfactory effects.

As such, one of the main reasons why cancer remains a refractory disease is known to be due to the expression of multidrug resistance (MDR) of cancer cells. Cancer cells that have become resistant to one drug are structurally resistant to completely different anticancer compounds, which is called multi-drug resistance. The development of such multi-drug resistance is the biggest obstacle in the treatment of metastatic cancer using chemotherapeutic agents. On the other hand, although several biological mechanisms causing multi-drug resistance have been reported, overexpression of P-glycoprotein (Pgp) is the most likely cause, and this has been proved biochemically and clinically. P-glycoprotein is a type of ATP-dependent transporter (ABC transporter) protein that has the function of releasing lipophilic foreign bodies in and out of cells. Many lipophilic drugs become substrates of P-glycoprotein, and most of them do not express the effect and are discharged out of cells by P-glycoprotein. Therefore, suppressing the expression of P-glycoprotein makes cancer cells having multidrug resistance in response to anticancer drugs and induces apoptosis, which is thought to be the most efficient way to overcome multidrug resistant cancer. Using P-glycoprotein-induced cells to search for compounds that induce apoptosis through co-administration with anticancer agents, search for inhibitors of P-glycoprotein or use fluorescent materials that bind to P-glycoprotein. P-glycoprotein inhibitors can be explored and identified.

Looking more closely at the P-glycoprotein, P-glycoprotein is a membrane protein encoded by 170 kDa by the human ABCB-1 (MDR1) gene, which is divided into six membranes and one It belongs to the superfamily of ATP binding cassettes (ABP) transport proteins composed of 1,280 amino acids formed of two homologous hydrophobic sites with ATP-binding sites. Expression of multidrug resistance is indicative of multidrug resistance through the mechanism by which P-glycoproteins bind directly to the drug and release the drug out of the cell by an energy dependent mechanism that consumes ATP. In addition, most human tumors, such as colon or kidney cancer, express P-glycoprotein and increase the expression rate, thereby exacerbating tumor deterioration.

Anticancer agents affected by P-glycoprotein include vinblastine, vincristine and navelbine of the vinca alkaloids family; Taxanes family paclitaxel (TAX), taxotere; Anthracyclines family of doxorubicin, daunorubicin, epirubicin and idarubicin; Epipodophyllotoxins family drugs etoposide and teniposide; Other drugs include colchicine, mitoxantrone, dactinomycin, topotecan, trimetrexate, mitramycin, mitomycin C Various drugs are known.

Therefore, such multi-drug resistance is an important limiting factor in the use of anticancer drugs, and clinically, the treatment rate of patients with cancer expressing P-glycoprotein is significantly lower than that of cancer patients who do not express it. Is showing.

There are various studies to overcome such multi-drug resistance, and many P-glycoprotein inhibitors have been developed so far.

First, the first-generation drugs, such as verapamil, calmodulin antagonists, steroidal drugs, immunosuppressants (cyclosporine) and antimalarial drugs (quinine), which are calcium channel inhibitors, are being used for other therapeutic purposes. Among the drugs are drugs that inhibit P-glycoprotein. However, first-generation drugs have not been sufficiently inhibited, and their original pharmacological action has been a side effect and failed to be used in clinical practice.

Second-generation drugs compensate for the shortcomings of the first generation, but many second-generation drugs not only inhibit other types of transporters but also inhibit metabolism-related enzymes, which increases the blood concentration of anticancer drugs for a long time. Showed a problem of increasing.

Third-generation drugs are highly selective for other transporters, have a very high inhibitory effect on P-glycoproteins, and are not metabolized by CYP450 3A4, which does not change the pharmacokinetic characteristics of anticancer drugs administered in combination, resulting in side effects from anticancer drugs. I never do that. Despite these characteristics, many of the third generation drugs that have been developed so far (Tariquidar, Zosuquidar, VX-710, Laniquidar, ONT-093, etc.), except for those in clinical trials, are cells that are caused by P-glycoprotein inhibitors themselves. Unexpected side effects from toxicity are a problem.

Therefore, there is a need for the development of P-glycoprotein inhibitors that do not have their own cytotoxicity.

Accordingly, the present inventors have studied to search for compounds having no self-cytotoxicity while inhibiting the overexpression of P-glycoprotein, which is a major cause of multi-drug resistance, and increasing the therapeutic efficiency of existing anticancer drugs, The present invention was synthesized, and it was confirmed that some of these compounds and known coumarin-based compounds exhibited superior inhibitory activity in P-glycoprotein expression.

An object of the present invention is to provide a novel coumarin-based compound.

Another object of the present invention is to provide a method for preparing the novel coumarin compound.

Another object of the present invention to provide a pharmaceutical composition for inhibiting multi-drug resistance containing the novel coumarin-based compound as an active ingredient.

In order to achieve the above object, the present invention provides a novel coumarin-based compound.

The present invention also provides a method for producing the novel coumarin compound.

Furthermore, the present invention provides a pharmaceutical composition for inhibiting multi-drug resistance containing the novel coumarin-based compound as an active ingredient.

The novel coumarin-based compounds according to the present invention exhibit an inhibitory activity against overexpression of P-glycoprotein, which is a major cause of multi-drug resistance, and an increase in anti-cancer activity upon co-administration with anti-cancer agents, thereby overcoming multi-drug resistance over-the-counter, therapeutic or It can be used as a modulator, enhance the effectiveness of anticancer drugs, increase the survival rate of cancer patients, reduce the dose of anticancer drugs administered in combination, reduce side effects on normal cells, and can provide economic benefits as well. It can be usefully used for treatment.

EMBODIMENT OF THE INVENTION Hereinafter, this invention is demonstrated in detail.

The present invention provides a novel coumarin compound represented by the following formula (1).

In Chemical Formula 1,

R is -X-Y,

X is C ₁ -C ₆ alkylene or carbonyl;

Y is phenyl or pyridyl, wherein the phenyl or pyridyl may be unsubstituted or substituted with one or more nitro, trifluoromethyl, halogen, C ₁ -C ₆ alkoxy or phenyl,

Preferably,

X is C ₁ -C ₂ alkylene or carbonyl,

Y is wherein Y is phenyl or pyridyl, the phenyl or pyridyl may be unsubstituted or substituted with one or more nitro, trifluoromethyl, halogen, C ₁ -C ₆ alkoxy or phenyl,

More preferably,

X is C ₁ -C ₂ alkylene,

Y is phenyl or pyridyl, wherein phenyl or pyridyl may be unsubstituted or substituted with one or more nitro or phenyl.

Specific examples of the compound of Formula 1 are as follows.

(1) 3-nitro-benzoic acid 6- (3-methyl-but-2-enyl) -2-oxo-2H-chromen-7-yl ester;

(2) 3,5-dimethoxy-benzoic acid 6- (3-methyl-but-2-enyl) -2-oxo-2H-chromen-7-yl ester;

(3) 4-bromo-benzoic acid 6- (3-methyl-but-2-enyl) -2-oxo-2H-chromen-7-yl ester;

(4) 6- (3-methyl-but-2-enyl) -7-phenethyloxy-chromen-2-one;

(5) 6- (3-methyl-but-2-enyl) -7- (4-nitro-benzyloxy) -chromen-2-one;

(6) 7- (biphenyl-4-ylmethoxy) -6- (3-methyl-but-2-enyl) -chromen-2-one;

(7) 6- (3-methyl-but-2-enyl) -7- (3-trifluoromethyl-benzyloxy) -chromen-2-one;

(8) 6- (3-methyl-but-2-enyl) -7- (pyridin-2-ylmethoxy) -chromen-2-one.

Preferred examples of the compound,

6- (3-Methyl-but-2-enyl) -7- (4-nitro-benzyloxy) -chromen-2-one; or

6- (3-methyl-but-2-enyl) -7- (pyridin-2-ylmethoxy) -chromen-2-one.

Table 1 below shows the structures of specific examples of the coumarin-based compound of Formula 1 according to the present invention.

compound Structure (Structure of R in Formula 1)

One

2

3

4

5

6

7

8

The present invention includes all of the novel coumarin-based compounds represented by Formula 1, as well as pharmaceutically acceptable salts, possible solvates or hydrates and prodrugs that can be prepared therefrom.

The present invention is also a group consisting of known coumarin-based compounds such as marmesin, decursinol, decursinol, decursin, decursinol angelate and marmesin angelate. It provides a pharmaceutical composition for inhibiting multi-drug resistance, containing a coumarin-based compound selected from the above as an active ingredient.

The memesin (marmesin), decursinol (decursinol), decursin (decursin), decursinol angelate (decursinol angelate) and mamesin angelate (marmesin angelate) are each represented by the following formulas 2 to 6, The compounds may be prepared or purchased according to known production methods.

On the other hand, the present invention provides a method for producing a novel coumarin compound of formula (1).

In Chemical Formula 1, a method for preparing a compound of the present invention corresponding to the case where R is -X-Y and X is carbonyl, as shown in Scheme 1 below,

It comprises the step of preparing a coumarin-based compound of Formula 1a through the esterification reaction of the starting material dimethylsuberosin (dimethylsuberosin) of the formula (7).

(In Scheme 1, Y is as defined in Formula 1, Z is OH, halide or OC (O) Y.)

The esterification reaction of Scheme 1 may be completed using a reagent for a conventional esterification reaction.

In one embodiment of the esterification reaction, the esterification reaction of Scheme 1 is achieved using acyl halide and base, or carboxylic anhydride.

On the other hand, another embodiment of the esterification reaction of Scheme 1 can also be achieved by reacting the carboxylic acid with dimethyl subsurosine of the formula (7) in the presence of a condensing agent and an amine catalyst.

As the condensing agent, dicyclohexylcarbodiimide (DCC), 1,1-carbonyldiimidazole, etc. may be used, and as the amine catalyst, trialkyl including pyridine, dimethylaminopyridine (DMAP), and triethylamine An amine etc. can be used, Preferably, dicyclohexyl carbodiimide can be used as a condensing agent, and dimethylamino pyridine can be used as an amine catalyst.

Dichloromethane, chloroform, tetrahydrofuran, etc. may be used as a reaction solvent of the esterification reaction of Scheme 1, and preferably dichloromethane may be used.

The esterification reaction of Scheme 1 may be achieved by stirring at room temperature for 12 hours to 24 hours.

In the general formula 1, wherein R is -XY and X is a C ₁ -C ₆ alkylene method for producing a compound of the present invention as shown in Scheme 2,

Preparing a coumarin compound of Formula 1b by reacting a starting material of dimethylsuberosin of Formula 7 with a halide compound represented by Y-X-Hal in the presence of a base.

(In Scheme 2, Hal is one of the halogen elements, X is alkylene of C ₁ -C ₆ , Y is as defined in formula (1).)

In this case, an aprotic polar solvent such as DMF or DMSO may be used as the reaction solvent, and preferably DMF may be used.

As the base in Scheme 2, sodium carbonate, potassium carbonate, calcium carbonate, sodium hydroxide, potassium hydroxide, calcium hydroxide and the like can be used.

The reaction can be made by stirring at room temperature for 12 to 24 hours.

On the other hand, dimethyl suverosine, which is a starting material of Scheme 1 or 2, may be prepared as shown in Scheme 3 below.

(In Reaction Scheme 3, Hal is one of the halogen elements, Bn represents benzyl and Boc represents t-butyloxycarbonyl.)

Method for preparing dimethyl subsurin is shown in Scheme 3,

Preparing a protected benzaldehyde of Chemical Formula 9 by reacting di-t-butyl dicarbonate in the presence of a base with benzaldehyde of Chemical Formula 8 as a starting material (Step 1);

Reducing the protected benzaldehyde of Formula 9 obtained in step 1 with a reducing agent to prepare a protected benzyl alcohol of Formula 10 (step 2);

Preparing a compound of formula 11 by reacting the protected benzyl halide of formula 10 obtained in step 2 with a Grignard reagent of 2-methyl-1-propenyl (step 3);

Preparing a compound of formula 12 by reacting the compound of formula 11 obtained in step 3 with the benzyl halide reagent in the presence of a base (step 4);

Treating the compound of Formula 12 obtained in step 4 under acidic conditions to remove the protective group Boc to prepare a compound of Formula 13 (step 5);

Reacting the compound of formula 13 obtained in step 5 with triehtylorthoformate (CH (OEt) ₃ ) in the presence of AlCl ₃ catalyst to obtain a compound of formula 14 (step 6);

Reacting (carbethoxymethylene) triphenylphosphorane with the compound of Formula 14 obtained in step 6 to obtain a compound of Formula 15 (step 7); And

The compound of Formula 15 obtained in step 7 is prepared using a reducing agent capable of removing the benzyl group to prepare dimethylsuberosine of formula 7 (step 8).

Hereinafter, a method for preparing dimethyl subsurin according to Scheme 3 will be described in detail.

Step 1 is a step of preparing a protected benzaldehyde of Chemical Formula 9 by reacting di-t-butyl daacarbonate (Boc ₂ O) with benzaldehyde of Chemical Formula 8 in the presence of a base. As a base, an inorganic base can be used preferably. Inorganic bases include potassium carbonate, sodium carbonate, calcium carbonate, sodium hydroxide, potassium hydroxide and the like.

At this time, as the reaction solvent, diethyl ether, tetrahydrofuran, dichloromethane, chloroform and the like can be used.

Step 2 is a step of reducing the protected benzaldehyde of Formula 9 obtained in Step 1 with a reducing agent to prepare a protected benzyl alcohol of Formula 10. The reducing agent may be any kind of reducing agent capable of reducing aldehyde to alcohol, and PCC, LAH, NaBH ₄ , a metal catalyst in the presence of hydrogen, sodium in the presence of alcohol, and the like are preferable.

Preferred examples of the reducing agent include NaBH ₄ , and the reaction solvent may be a mixture of tetrahydrofuran, diethyl ether, tetrahydrofuran and water.

Step 3 is a step of preparing a compound of Formula 11 by reacting the Grignard reagent of 2-methyl-1-propenyl with the protected benzyl halide of Formula 10 obtained in Step 2. As a Grignard reagent of 2-methyl-1-propenyl, 2-methyl-1-propenyl magnesium bromide or 2-methyl-1-propenyl magnesium chloride is preferably used, and as reaction solvent, diethyl ether or Tetrahydrofuran can be used preferably.

Step 4 is a step of preparing a compound of formula 12 by reacting the compound of formula 11 obtained in step 3 with the benzyl halide reagent in the presence of a base. Benzyl halide reagents are preferably benzyl bromide and benzyl chloride, and inorganic bases may be used as the base, and inorganic bases include potassium carbonate, sodium carbonate, calcium carbonate, sodium hydroxide, potassium hydroxide and the like.

Step 5 is a step of preparing a compound of Formula 13 by removing the Boc group as a protective group by treating the compound of Formula 12 obtained in step 4 under acidic conditions. Any acidic condition capable of removing the Boc group is possible, and preferably, an acidic condition including hydrochloric acid is used.

Step 6 is a step of obtaining a compound of formula 14 by reacting the compound of formula 13 obtained in step 5 with triethylorthoformate (CH (OEt) ₃ ) in the presence of AlCl ₃ catalyst. As the reaction solvent, benzene, toluene, xylene or the like can be preferably used.

Step 7 is a step of obtaining a compound of Formula 15 by reacting (carbethoxymethylene) triphenylphosphorane with the compound of Formula 14 obtained in Step 6. As the reaction solvent, N, N-diethylaniline can be preferably used.

Step 8 is a step of preparing dimethyl suverosine of the formula (7) using a reducing agent capable of removing the benzyl group of the compound of formula (15) obtained in step 7. As the reducing agent, a metal catalyst such as Raney Ni may be used in the presence of hydrogen.

The novel coumarin-based compound according to the present invention is not limited to the above-mentioned preparation method, and may be used as long as it is a method capable of synthesizing the novel coumarin-based compound even in a known method as well as an already known method.

As described above, the novel coumarin-based compound prepared according to the present invention may be separated and purified by high-performance liquid chromatography after preparation, and then the molecular structure may be confirmed by nuclear magnetic resonance.

Furthermore, the present invention provides a pharmaceutical composition for inhibiting multi-drug resistance, which contains a coumarin compound of Formula 1 as an active ingredient.

Compounds of Formula 1 of the present invention can control or treat multi-drug resistance by inhibiting overexpression of a protein that induces multi-drug resistance to anticancer drugs, wherein the protein that induces multi-drug resistance is a P-glycoprotein expressed in cancer cells To be the target.

The pharmaceutical composition containing the coumarin-based compound of the present invention as an active ingredient inhibits the induction of multi-drug resistance by coadministration with an anticancer agent, thereby enhancing the anticancer effect of the anticancer agent, and also reducing the dose of the anticancer agent coadministered with the anticancer agent alone. Reduce side effects on normal cells

Multidrug resistance is mainly due to overexpression of P-glycoprotein by MDR1 in the cell membrane of cancer cells, which directly binds to anticancer drugs, resulting in the release of the drug out of the cell and reducing intracellular drug accumulation. As the drug concentration inside the cancer cells is reduced. Therefore, compounds that inhibit the overexpression of P-glycoprotein have the effect of inhibiting multi-drug resistance.

The compound of the present invention enhances the cytotoxicity of paclitaxel against MES-SA / DX5 cancer cell line, which is a P-glycoprotein expressing cell line treated with paclitaxel, inhibits the expression of P-glycoprotein (see Table 2), and is a conventional anticancer agent. Compared to the IC ₅₀ values measured with anti-cancer agents alone when combined with paclitaxel, the IC ₅₀ values were lowered by up to 36-fold for the MES-SA / DX5 cell line. This shows superior multi-drug resistance inhibitory activity than verapamil used as a conventional multi-drug resistance inhibitor (see Table 2). Therefore, the compounds of the present invention effectively inhibit P-glycoprotein that causes multi-drug resistance, and thus can be usefully used to treat or modulate multi-drug resistance.

Meanwhile, the compounds of the present invention are not only cancers in which P-glycoprotein is normally expressed, such as colon cancer, rectal cancer, bladder cancer, menstrual cancer, breast cancer, lung cancer, etc., but also acute myeloid leukemia (AML) in which the expression of P-glycoprotein is not high. ), And can be used to treat or regulate multi-drug resistance by anticancer agents in malignant lymphomas and the like.

In particular, the compounds of the present invention include vinblastine, vincristine and navelbine of the vinca alkaloids family; Taxanes family of paclitaxel (TAX), taxotere; Anthracyclines family of doxorubicin, daunorubicin, epirubicin and idarubicin; Epipodophyllotoxins family drugs etoposide and teniposide; Other drugs include colchicine, mitoxantrone, dactinomycin, topotecan, trimetrexate, mithramycin, mitomycin C It can be used to treat or control multi-drug resistance to anticancer agents.

When the composition of the present invention is used as a medicine, the pharmaceutical composition containing the coumarin-based compound represented by Formula 1 as an active ingredient may be formulated and administered in various oral or parenteral dosage forms as described below during clinical administration. It is not limited to this.

Formulations for oral administration include, for example, tablets, pills, hard / soft capsules, solutions, suspensions, emulsifiers, syrups, granules, elixirs, and the like. Rose, sucrose, mannitol, sorbitol, cellulose and / or glycine), lubricants such as silica, talc, stearic acid and its magnesium or calcium salts and / or polyethylene glycols. Tablets may also contain binders such as magnesium aluminum silicate, starch paste, gelatin, methylcellulose, sodium carboxymethylcellulose and / or polyvinylpyrrolidine, and optionally such as starch, agar, alginic acid or its sodium salt. Disintegrant or boiling mixtures and / or absorbents, colorants, flavors, and sweeteners.

Pharmaceutical compositions comprising the compound represented by Formula 1 as an active ingredient may be administered parenterally, and parenteral administration may be by injecting subcutaneous injection, intravenous injection, intramuscular injection, or intrathoracic injection.

In this case, the salt of the coumarin-based compound of Formula 1 may be prepared as a solution or a suspension by mixing with a stabilizer or a buffer in water to formulate into a parenteral formulation, it may be prepared in ampule or vial unit dosage form. The compositions may contain sterile and / or preservatives, stabilizers, hydrating or emulsifying accelerators, auxiliaries such as salts and / or buffers for the control of osmotic pressure, and other therapeutically useful substances, and conventional methods of mixing, granulating It may be formulated according to the formulation or coating method.

In addition, the dosage of the compound of the present invention to the human body may vary depending on the age, weight, sex, dosage form, health condition and degree of disease of the patient, and generally based on an adult patient having a weight of 70 kg. It is 30-120 mg / day, Preferably it is 40-80 mg / day, It can also divide and administer once a day to several times at regular time intervals according to the judgment of a doctor or a pharmacist.

Hereinafter, the present invention will be described in detail through Examples and Experimental Examples. However, the following Examples and Experimental Examples are merely illustrative of the present invention, and the content of the present invention is not limited by the following Examples.

Silica gel 60 (230-400 mesh Kieselgel 60) was used for flash column chromatography, unless otherwise specified in the present preparations and examples, and in preparative thin layer chromatography (TLC), a silica gel plate of a glass substrate ( 1 mm thick), and a 0.25 mm thick silica plate (E. Merck, silica gel 60 F254) was used for TLC to confirm the progress of the reaction.

Preparation Example Preparation of Dimethyl Subrosine

(Step 1): Preparation of 2,4-di-t-butyl 1-formylphenyl carbonate

Starting materials 2,4-dihydroxybenzaldehyde (2.0 g, 14.5 mmol), di-tert-butyl dicarbonate (Boc ₂ O; 12.6 g, 57.9 mmol) and potassium carbonate (4.0 g, 28.9 mmol) Put in ethyl ether (60 ml) and stir at room temperature to dissolve completely. After filtering the reaction mixture, the mixed filtrate obtained by washing the precipitate with diethyl ether was distilled under reduced pressure. Water was added to the obtained residue to terminate the reaction, and the resultant was separated with ethyl acetate. The ethyl acetate layer was separated, washed with brine and water, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The obtained residue was purified by flash column chromatography (ethyl acetate: n-hexane = 2: 1) to obtain 4.4 g (yield 90%) of the title compound.

¹ H NMR (CDCl 3, 300 MHz) δ 10.12 (s, 1H), 7.88 (d, J = 7.8 Hz, 1H), 7.23 (d, J = 1.8 Hz, 1H), 7.20 (s, 1H), 1.55 ( d, J = 3 Hz, 18H).

(Step 2): Preparation of 2,4-di-t-butyl 1-hydroxymethylphenyl carbonate

THF: To a solution of the compound obtained in step 1 (130 mg, 384 mmol) dissolved in water (19: 1, 1 ml) was added NaBH ₄ (15.3 mg, 403 mmol, contained in 0.27 ml of water) at 0 ° C. Stir for 15 minutes. The reaction mixture is poured into iced water with ice and then extracted three times with diethylether. The combined diethyl ether layers were washed with brine and water, dried over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The obtained residue was purified by prep TLC (ethyl acetate: n-hexane = 3: 7) to obtain 93.7 mg (yield 70%) of the title compound.

¹ H NMR (CDCl ₃ , 300 MHz) δ 7.42 (d, J = 8.4 Hz, 1H), 7.07 (d, J = 2.4 Hz, 1H), 7.04 (dd, J = 2.1, 3.9 Hz, 1H), 4.56 (s, 2H), 2.37 (brs, 1 H), 1.52 (s, 18 H).

(Step 3): Preparation of t-butyl 3-hydroxy-4- (3-methylbut-2-enyl) phenyl carbonate

To methyl THF (21 ml) solution of the compound (1.29 g, 3.81 mmol) obtained in step 2 was added 2-methyl-1-propenyl magnesium bromide (11.44 mmol, 0.5 M in THF 22.89 ml) at 0 ° C. and stirred for 1 hour. do. The reaction mixture is allowed to warm to room temperature and then stirred for a further 10 hours. The reaction mixture was poured into 0.5 N HCl aqueous solution to terminate the reaction, followed by extraction with diethyl ether. The extracted diethyl ether layer was washed with brine and water, dried over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The obtained residue was purified by flash column chromatography (ethyl acetate: n-hexane = 3: 7) to obtain 760 mg (yield 70%) of the title compound.

¹ H NMR (CDCl 3, 300 MHz) δ 7.04 (d, J = 8.7 Hz, 1H), 6.64 (dd, J = 2.1, 8.4 Hz, 1H), 6.57 (d, J = 1.8 Hz, 1H), 5.84 ( s, 1H), 5.26 (m, 1H), 3.26 (d, J = 7.2 Hz, 2H), 1.74 (s, 3H), 1.72 (s, 3H), 1.55 (s, 9H);

MS (ESI) m / z 301 (M + Na) < + >.

(Step 4): Preparation of t-butyl 3-benzyloxy-4- (3-methylbut-2-enyl) phenyl carbonate

The suspension of compound (0.60 g, 2.16 mmol), benzylbromide (0.44 g, 2.59 mmol) and potassium carbonate DMF (10 ml) obtained in step 3 was stirred at room temperature for 12 hours. The reaction mixture was poured into ice-cold water and extracted with ethyl acetate. The combined extracts were washed with brine and water. The ethyl acetate layer was washed with brine and water, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The obtained residue was purified by flash column chromatography (ethyl acetate: n-hexane = 1: 9) to obtain 650 mg (yield 81.4%) of the title compound.

¹ H NMR (CDCl ₃ , 300 MHz) δ 7.38 (m, 5H), 7.14 (d, J = 8.4 Hz, 1H), 6.75 (s, 1H), 6.73 (d, J = 1.8 Hz, 1H), 5.31 (m, 1H), 5.06 (s, 2H), 3.35 (d, J = 7.5 Hz, 2H), 1.74 (s, 3H), 1.65 (s, 3H), 1.57 (s, 9H);

¹³ C NMR (CDCl ₃ , 75 MHz) δ 156.80, 152.01, 149.88, 136.90, 132.59, 129.49, 128.49, 127.94, 127.86, 127.31, 122.27, 113.06, 105.28, 83.38, 70.07, 28.25, 27.72, 25.75, 17.72;

MS (ESI) m / z 391 (M + Na) ⁺ .

(Step 5): Preparation of 3-benzyloxy-4- (3-methylbut-2-enyl) phenol

The n-butanol (200 ml) solution of the compound (2.00 g, 5.43 mmol) obtained in step 4 was added to 4 M HCl dioxane solution (70 ml) at room temperature, followed by TLC, followed by stirring until the reaction was completed. The reaction mixture was quenched with water cooled to ice, neutralized with saturated aqueous sodium hydrogen carbonate solution, and extracted with a methanol: dichloromethane (1: 9) mixture. The extracted organic layer was washed with brine and water, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The obtained residue was purified by column chromatography (ethyl acetate: n-hexane = 1.5: 8.5) to obtain 1.32 g (yield 90.6%) of the title compound.

¹ H NMR (CDCl ₃ , 300 MHz,) δ 7.36 (m, 5H), 6.99 (d, J = 8.1 Hz, 1H), 6.45 (d, J = 2.4 Hz, 1H), 6.35 (dd, J = 2.4 , 8.1 Hz, 1H), 5.29 (m, 1H), 5.04 (s, 2H), 4.55 (brs, 1H), 3.29 (d, J = 7.5 Hz, 2H), 1.72 (s, 3H), 1.65 (s , 3H);

¹³ C NMR (CDCl ₃ , 75 MHz) δ 157.29, 154.65, 137.17, 132.06, 129.82, 128.48, 127.76, 127.22, 127.14, 122.94, 122.75, 106.85, 100.09, 69.90, 28.01, 25.77, 17.70;

MS (ESI) m / z 269 (M + H) ⁺ .

(Step 6): Preparation of 4-benzyloxy-2-hydroxy-5- (3-methylbut-2-enyl) benzaldehyde

AlCl ₃ (391 mg, 2.93) in a benzene (25 ml) solution of the compound (523 mg, 1.95 mmol) and triethylorthoformate (CH (OEt) ₃ ; 1.45 g, 9.75 mmol) obtained in step 5; mmol) was added at room temperature and stirred for 30 minutes. The reaction mixture was lowered to 0 ° C and 3 M HCl aqueous solution (28 ml) was added dropwise. The reaction mixture was warmed to room temperature and then extracted with diethyl ether and ethyl acetate. The combined organic layers were washed with brine and water, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The obtained residue was purified by flash column chromatography (ethyl acetate: n-hexane = 1: 9) to obtain 280 mg (yield 48.4%) of the title compound.

¹ H NMR (CDCl ₃ , 300 MHz) δ 11.45 (s, 1H), 9.72 (s, 1H), 7.42 (m, 5H), 7.27 (d, J = 2.4 Hz, 1H), 5.30 (m, 1H) , 5.14 (s, 2H), 3.31 (d, J = 7.2 Hz, 2H), 1.78 (s, 3H), 1.66 (s, 3H);

¹³ C NMR (CDCl ₃ , 75 MHz) δ 194.54, 163.67, 163.08, 135.88, 133.46, 133.41, 128.66, 128.21, 127.27, 123.06, 121.62, 114.50, 99.71, 70.32, 27.71, 25.79, 17.74;

MS (ESI) 297 (M + H) ⁺ , 319 (M + Na) ⁺ .

(Step 7): Preparation of 7-benzyloxy-6- (3-methylbut-2-enyl) -2H-chromen-2-one

The compound obtained in step 6 and (carbethoxymethylene) triphenylphosphorane ((carbethoxymethylene) triphenylphosphorane); 0.44 g, 1.27 mmol) of N, N-diethylaniline (21 ml) solution was heated to 190 ° C. for 5 hours in an argon atmosphere. The reaction solution was cooled and poured into 1 liter of 1.5 M HCl aqueous solution, and extracted three times with ethyl acetate. The combined ethyl acetate extracts were washed with 1.5 M HCl aqueous solution and brine, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The obtained solid was purified by flash column chromatography (ethyl acetate: n-hexane = 3: 7) to obtain 290 mg (yield 88.8%) of the title compound.

¹ H NMR (CDCl ₃ , 300 MHz) δ 7.66 (d, J = 9.6 Hz, 1H), 7.50-7.38 (m, 5H), 7.31 (s, 1H), 6.88 (s, 1H), 6.28 (d, J = 9.6 Hz, 1H), 5.35 (m, 1H), 5.2 (s, 2H), 3.43 (d, J = 7.2 Hz, 2H), 1.82 (s, 3H), 1.71 (s, 3H);

¹³ C NMR (CDCl ₃ , 75 MHz) δ 161.48, 159.57, 154.31, 143.57, 135.98, 133.63, 128.69, 128.22, 127.81, 127.60, 127.26, 121.42, 112.91, 112.10, 99.79, 70.39, 28.12, 25.80, 17.77;

MS (ESI) m / z 321 (M + H) ⁺ , 343 (M + Na) ⁺ .

(Step 8): Preparation of Dimethylsubrosine

An ethanol (6 ml) solution of the compound obtained in step 7 was added to an aqueous slurry of Raney nickel (0.88 g), and the mixture was stirred at room temperature in an argon atmosphere for 3 hours. The reaction mixture was quickly filtered through a plug of celite and then concentrated under reduced pressure. The residue was purified by flash column chromatography (ethyl acetate: n-hexane = 3: 7) to obtain 160 mg (yield 89.6%) of a dimethylsuberosine compound.

¹ H NMR (CDCl ₃ , 300 MHz) δ 7.65 (d, J = 9 Hz, 1H), 7.58 (s, 1H), 7.19 (s, 1H), 7.05 (s, 1H) 6.23 (d, J = 9.9 Hz, 1H), 5.32 (m, 1H), 3.37 (d, J = 7.2 Hz, 2H), 1.78 (s, 3H), 1.74 (s, 3H);

¹³ C NMR (CDCl ₃ , 75 MHz) δ 162.20, 158.42, 154.15, 144.12, 135.16, 128.31, 125.56, 120.96, 112.39, 112.31, 103.22, 28.52, 25.81, 17.87;

MS (ESI) m / z 231 (M + H) ⁺ , 253 (M + Na) ⁺ .

EXAMPLES Preparation of New Coumarin-Based Compound

Example 1 Preparation of 3-nitro-benzoic acid 6- (3-methyl-but-2-enyl) -2-oxo-2H-chromen-7-yl ester

3-nitrobenzoyl chloride (27 mg, 0.14 mmol) and triethylamine (0.04) were cooled down with a dichloromethane (3 ml) solution of dimethyl suverosine (30 mg, 0.13 mmol) obtained in the above preparation. ml, 0.26 mmol) was added dropwise. The reaction mixture was stirred at room temperature for 18 hours, the reaction was terminated with saturated aqueous sodium hydrogen carbonate solution, extracted with dichloromethane, the dichloromethane solution was dried over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The obtained residue was purified by flash column chromatography (ethyl acetate: n-hexane = 1: 5) to obtain 18.6 mg (yield 62%) of the title compound.

¹ H NMR (CDCl ₃ , 300 MHz) δ 9.04-9.03 (m, 1H), 8.55-8.51 (m, 1H), 7.79-7.69 (m, 3H), 7.39 (s, 1H), 7.19 (s, 1H ), 6.42 (d, J = 9.3 Hz, 1H), 5.21 (t, J = 6.6 Hz, 1H), 3.32 (d, J = 6.6 Hz, 2H), 1.69 (s, 3H), 1.57 (s, 3H );

MS (ESI) m / z 380 (M + H) ⁺ .

Example 2 Preparation of 3,5-dimethoxy-benzoic acid 6- (3-methyl-but-2-enyl) -2-oxo-2H-chromen-7-yl ester

Dimethylsubrosine (30 mg, 0.13 mmol), 3,5-dimethoxy benzoic acid (28 mg, 0.16 mmol), DCC (34 mg, 0.16 mmol) and DMAP (130 mg, 1.06 mmol) obtained in the above preparation Was dissolved in 5 ml of anhydrous dichloromethane and stirred at room temperature for 24 hours. The residue obtained after evaporation of the solvent was purified by flash column chromatography (MeOH: dichloromethane = 1:50) to give 15.9 mg (yield 53%) of the title compound.

¹ H NMR (CDCl ₃ , 300 MHz) δ 7.69 (d, J = 9.6 Hz, 1H), 7.36-7.33 (m, 3H), 7.17 (s, 1H), 6.75-6.74 (m, 1H), 6.39 ( d, J = 9.6 Hz, 1H), 5.22 (t, J = 7.2 Hz, 1H), 3.87 (s, 6H), 3.31 (d, J = 7.2 Hz, 2H), 1.71 (s, 3H), 1.59 ( s, 3H);

MS (ESI) m / z 395 (M + H) ⁺ .

Example 3 Preparation of 4-Bromo-benzoic acid 6- (3-methyl-but-2-enyl) -2-oxo-2H-chromen-7-yl ester

4-bromobenzoyl chloride (33.26 mg, 0.15 mmol) and triethylamine (0.04 ml, 0.28 mmol) were cooled in a dichloromethane solution of dimethyl submerosine (31.7 mg, 0.14 mmol) obtained in the above preparation with ice. ) Was added to the mixture. The reaction mixture was stirred at room temperature for 18 hours, the reaction was terminated with saturated aqueous sodium hydrogen carbonate solution, extracted with dichloromethane, the dichloromethane solution was dried over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The obtained residue was purified by flash column chromatography (ethyl acetate: n-hexane = 1: 5) to obtain 23.4 mg (yield 74%) of the title compound.

¹ H NMR (CDCl ₃ , 300 MHz) δ 8.08-8.04 (m, 1H), 7.68 (m, 4H), 7.36 (s, 1H), 7.17 (s, 1H), 6.40 (d, J = 9.6 Hz, 1H), 5.21 (t, J = 7.5 Hz, 1H), 3.30 (d, J = 7.5 Hz, 2H), 1.69 (s, 6H);

MS (ESI) m / z 413 (M + H) ⁺ .

Example 4 Preparation of 6- (3-methyl-but-2-enyl) -7-phenethyloxy-chromen-2-one

2-bromoethylbenzene (0.26 mmol) was added while stirring a solution of DMF (3 ml) of dimethyl suverosine (40 mg, 0.18 mmol) and potassium carbonate (73 mg, 0.53 mmol) obtained in the above preparation at room temperature. It was. The reaction mixture was stirred for 18 hours at room temperature. When the reaction was completed, water was added and the mixture was extracted twice with ethyl acetate. The ethyl acetate extract was washed with water and brine, dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by flash column chromatography (ethyl acetate: n-hexane = 1:10) to obtain 48 mg (yield 83%) of the title compound.

¹ H NMR (CDCl ₃ , 300 MHz) δ 7.59 (d, J = 9.6 Hz, 1H), 7.35-7.28 (m, 5H), 7.16 (s, 1H), 6.75 (s, 1H), 6.21J = 9.6 Hz, 1H), 5.24 (t, J = 7.2 Hz, 1H), 4.24 (t, J = 6.6 Hz, 2H), 3.28 (d, J = 7.2 Hz, 2H), 3.15 (t, J = 6.6 Hz, 2H), 1.75 (s, 3H), 1.68 (s, 3H);

MS (ESI) m / z 357 (M + Na) ⁺ , 335 (M + H) ⁺ .

Example 5 Preparation of 6- (3-Methyl-but-2-enyl) -7- (4-nitro-benzyloxy) -chromen-2-one

4-nitrobenzylbromide (0.13 mmol) was added to the solution of DMF (2 ml) of dimethyl suverosine (30 mg, 0.19 mmol) and potassium carbonate (55 mg, 0.39 mmol) obtained in the above preparation at room temperature. It was. The reaction mixture was stirred for 18 hours at room temperature. When the reaction was completed, water was added and the mixture was extracted twice with ethyl acetate. The ethyl acetate extract was washed with water and brine, dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by flash column chromatography (ethyl acetate: n-hexane = 1: 5) to obtain 39.5 mg (yield 82%) of the title compound.

¹ H NMR (CDCl ₃ , 300 MHz) δ 8.26 (d, J = 8.27 Hz, 2H), 7.62 (d, J = 7.6 Hz, 2H), 7.80-6.45 (m, 4H), 5.82 (s, 2H) , 5.20 (t, J = 9.6 Hz, 1H), 3.22 (d, J = 7.2 Hz, 2H), 1.75 (s, 3H), 1.68 (s, 3H);

MS (ESI) m / z 366 (M + H) ⁺ .

Example 6 Preparation of 7- (biphenyl-4-ylmethoxy) -6- (3-methyl-but-2-enyl) -chromen-2-one

4-Broethylbiphenyl (0.19 mmol) was prepared by stirring a solution of DMF (2 ml) of dimethyl suverosine (30 mg, 0.19 mmol) and potassium carbonate (55 mg, 0.39 mmol) obtained in the above preparation at room temperature. Was added. The reaction mixture was stirred for 18 hours at room temperature. When the reaction was completed, water was added and the mixture was extracted twice with ethyl acetate. The ethyl acetate extract was washed with water and brine, dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by flash column chromatography (ethyl acetate: n-hexane = 1: 5) to obtain 40.3 mg (yield 78%) of the title compound.

¹ H NMR (CDCl ₃ , 300 MHz) δ 7.80 (d, J = 9.6 Hz, 1H), 7.40-7.28 (m, 9H), 7.14 (s, 1H), 6.59 (s, 1H), 6.45 (d, J = 9.6 Hz, 1H), 5.80 (s, 2H), 5.20 (t, J = 6.6 Hz, 1H), 3.22 (d, J = 7.2 Hz, 2H), 1.75 (s, 3H), 1.68 (s, 3H);

MS (ESI) m / z 367 (M + H) ⁺ , 419 (M + Na) ⁺ .

Example 7 Preparation of 6- (3-Methyl-but-2-enyl) -7- (3-trifluoromethyl-benzyloxy) -chromen-2-one

4- (trifluoromethyl) benzyl bromide (0.19) of DMF (2 ml) of dimethyl suverosine (30 mg, 0.19 mmol) and potassium carbonate (55 mg, 0.39 mmol) obtained in the above preparation was stirred at room temperature. mmol) was added. The reaction mixture was stirred for 18 hours at room temperature. When the reaction was completed, water was added and the mixture was extracted twice with ethyl acetate. The ethyl acetate extract was washed with water and brine, dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by flash column chromatography (ethyl acetate: n-hexane = 1: 5) to obtain 47 mg (yield 92%) of the title compound.

¹ H NMR (CDCl ₃ , 300 MHz) δ 7.80 (d, J = 9.6 Hz, 1H), 7.32 (d, J = 9.6 Hz, 1H), 7.38-7.12 (m, 4H), 6.59 (s, 1H) , 6.45 (d, J = 9.6 Hz, 1H), 5.80 (s, 2H), 5.20 (t, J = 7.2 Hz, 1H), 3.22 (d, J = 7.2 Hz, 2H), 1.75 (s, 3H) , 1.68 (s, 3 H);

MS (ESI) m / z 389 (M + H) ⁺ .

Example 8 Preparation of 6- (3-Methyl-but-2-enyl) -7- (pyridin-2-ylmethoxy) -chromen-2-one

2-bromoethyl-pyridine (0.20 mmol) was prepared by stirring a solution of DMF (3 ml) of dimethyl suverosine (30 mg, 0.19 mmol) and potassium carbonate (55 mg, 0.39 mmol) obtained in the above preparation at room temperature. Was added. The reaction mixture was stirred for 18 hours at room temperature. When the reaction was completed, water was added and the mixture was extracted twice with ethyl acetate. The ethyl acetate extract was washed with water and brine, dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by flash column chromatography (ethyl acetate: n-hexane = 1: 2) to obtain 24 mg (yield 79%) of the title compound.

¹ H NMR (CDCl ₃ , 300 MHz) δ 8.65 (d, J = 4.2 Hz, 1H), 7.84 (t, J = 7.5 Hz, 1H), 7.63-7.56 (m, 2H), 7.36 (s, 1H) , 7.23 (s, 1H), 6.84 (s, 1H), 6.24 (d, J = 9.6 Hz, 1H), 5.36-5.29 (m, 3H), 3.42 (d, J = 3.43 Hz, 2H), 1.78 ( s, 3H), 1.70 (s, 3H);

MS (ESI) m / z 344 (M + Na) ⁺ , 322 (M + H) ⁺ .

Experimental Example Measurement of Multi-Drug Resistance Inhibitory Effect

(1) Cell culture

The MES-SA / DX5 (ATCC Number: CRL-1977) cell line was purchased from ATCC and used and cultured based on the culture method described by ATCC. MES-SA / DX5 cell lines were cultured in DMEM (or RPMI-1640) containing 10% FBS and passaged from 1: 6 to 1:10 every 2-3 days.

(2) measurement of multi-drug resistance inhibitory effect

In order to determine the multi-drug resistance inhibitory activity of the compound according to the present invention, the following experiment was performed.

MES-SA / DX5 cell lines were dispensed into 96-well plates and precultured for 24 hours to allow cells to adhere to the plate bottom face. The attached cells were treated with the control group (without multidrug resistance inhibitor) and the compound prepared in Examples 4, 5, and 8 (5.0 μM) according to the serial dilution concentration of the anticancer agent paclitaxel (Taxol), and Mamesine represented by the general formula (6). Angelate compounds and verapamil (5.0 μM) were added as a control group and incubated for 72 hours. After the incubation was completed, 0.01 mL of Cell Counting Kit-8 (Dojindo Corporate) solution was added to each well in each well, incubated in a cell incubator for 1 to 3 hours, and then absorbance was measured at 450 nm. Absorbance was measured using a microplate reader (Tecan), and 50% of cancer cells were shown in Table 2 by measuring the effective concentration (IC ₅₀ ) of paclitaxel in which growth was inhibited.

Experimental group
IC ₅₀ (μM) in paclitaxel

Resistance inhibitory effect
Control group (paclitaxel alone)

5.99
1.0
Example 4 Compound

1.93
3.1
Example 5 Compound

0.551
10.9
Example 8 Compound

0.166
36.0
Mamesin Angelate
(marmesin angelate)
1.03
5.8
Comparison
(Verapamil)
0.234
25.6

On the other hand, coumarin-based compounds and verapamil according to the present invention can be seen to enhance the cytotoxicity of paclitaxel against the MES-SA / DX5 cancer cell line P-glycoprotein expressing cell line. That is, in the case of the coumarin-based compound according to the present invention when 5.0 μM in combination with paclitaxel, it is shown that the IC ₅₀ value is lower than the paclitaxel alone treatment group for the MES-SA / DX5 cell line. In particular, in the case of the compound of Example 8, the IC ₅₀ value is lowered by 36 times compared with the paclitaxel alone treatment group, which shows superior drug resistance inhibitory activity than verapamil used as a conventional drug resistance inhibitor.

Therefore, the coumarin compound according to the present invention effectively inhibits P-glycoprotein that causes multi-drug resistance, and thus can be usefully used for treating or controlling multi-drug resistance. In addition, since no cytotoxicity of the compound of Example 8 was observed at the experimental concentration, it is expected that side effects due to self-cytotoxicity, which is a major problem of P-glycoprotein inhibitors, will be minimized.

On the other hand, the coumarin compound of Formula 1 according to the present invention can be formulated in various forms according to the purpose. The following are some examples of formulation methods containing the compound represented by Formula 1 according to the present invention as an active ingredient, but the present invention is not limited thereto.

Preparation Example 1 Preparation of Tablet

Active ingredient 100 mg

Corn starch 68 mg

Lactose 90 mg

Microcrystalline cellulose 40 mg

Magnesium Stearate 2mg

According to the conventional method of preparing tablets, the above ingredients were added to the indicated contents, uniformly mixed, stirred and granulated. After drying, tablets were prepared using a tablet press.

Preparation Example 2 Preparation of Capsule

Active ingredient 80 mg

Corn starch 68 mg

Lactose 90 mg

Microcrystalline cellulose 60 mg

Magnesium Stearate 2mg

According to a conventional method for preparing a capsule, the desired components were prepared by adding the above components in the amounts shown in the present invention, mixing them uniformly, and filling the gelatine capsules with an appropriate size.

Preparation Example 2 Preparation of Injection

50 mg of active ingredient

Sodium metabisulfite 1.5 mg

Methyl paraben 1.0 mg

Propyl Paraben 0.1 mg

Appropriate amount of purified water for injection

According to a conventional method for preparing an injectable drug, the above components are dissolved in boiling water with a given content, stirred, filled, cooled, and filled into a 2 ml sterile vial, and supplemented with an appropriate amount of injectable purified water to 2 ml. Prepared.

Claims (11)

Coumarin-based compounds represented by Formula 1 below: [Formula 1]

(In the formula 1, R is -X-Y, X is C ₁ -C ₆ alkylene or carbonyl; Y is phenyl or pyridyl, wherein the phenyl or pyridyl may be unsubstituted or substituted with one or more nitriles, trifluoromethyl, halogen, C ₁ -C ₆ alkoxy or phenyl.). According to claim 1, wherein in the definition of Formula 1 X is C ₁ -C ₂ Alkylene or carbonyl, Wherein Y is phenyl or pyridyl, wherein the phenyl or pyridyl may be unsubstituted or substituted with one or more nitriles, trifluoromethyl, bromo, methoxy or phenyl. 2. The compound of claim 1, wherein in the definition of Formula 1, X is C ₁ -C ₂ alkylene, Y is phenyl or pyridyl, and the phenyl or pyridyl is unsubstituted or used as one or more nitro or Coumarin-based compound which may be substituted with phenyl. The coumarin-based compound of claim 1, wherein (1) 3-nitro-benzoic acid 6- (3-methyl-but-2-enyl) -2-oxo-2H-chromen-7-yl ester; (2) 3,5-dimethoxy-benzoic acid 6- (3-methyl-but-2-enyl) -2-oxo-2H-chromen-7-yl ester; (3) 4-bromo-benzoic acid 6- (3-methyl-but-2-enyl) -2-oxo-2H-chromen-7-yl ester; (4) 6- (3-methyl-but-2-enyl) -7-phenethyloxy-chromen-2-one; (5) 6- (3-methyl-but-2-enyl) -7- (4-nitro-benzyloxy) -chromen-2-one; (6) 7- (biphenyl-4-ylmethoxy) -6- (3-methyl-but-2-enyl) -chromen-2-one; (7) 6- (3-methyl-but-2-enyl) -7- (3-trifluoromethyl-benzyloxy) -chromen-2-one; And (8) A coumarin compound, which is selected from the group consisting of 6- (3-methyl-but-2-enyl) -7- (pyridin-2-ylmethoxy) -chromen-2-one. The coumarin-based compound of claim 1, wherein 6- (3-Methyl-but-2-enyl) -7- (4-nitro-benzyloxy) -chromen-2-one; or A coumarin-based compound which is 6- (3-methyl-but-2-enyl) -7- (pyridin-2-ylmethoxy) -chromen-2-one. As shown in Scheme 1, a step of preparing a coumarin compound of Formula 1 (a) through an esterification reaction of dimethylsuberosin of Formula 7, which is a starting material, with an acyl halide in the presence of a base Method for preparing the coumarin compound of claim 1 comprising: Scheme 1

(In Scheme 1, Y is as defined in formula 1 of claim 1, Z is a halide.). As shown in Scheme 2, a step of preparing a coumarin compound of Formula 1 (b) by reacting dimethylsuberosin of Formula 7, which is a starting material, with a halide compound represented by YX-Hal in the presence of a base Method for producing a coumarin compound of claim 1 comprising: Scheme 2

(In Scheme 2, Hal is one of the halogen elements, X is C ₁ -C ₆ alkylene, Y is as defined in formula 1 of claim 1). A pharmaceutical composition for inhibiting multi-drug resistance, comprising the coumarin-based compound of any one of claims 1 to 5 as an active ingredient. The pharmaceutical composition for inhibiting multi-drug resistance according to claim 8, wherein the coumarin-based compound inhibits over-expression of P-glycoprotein causing multi-drug resistance to anti-cancer agents. According to claim 8, wherein the anticancer agent (vinblastine, vincristine, navelbine, paclitaxel (TAX), taxotere, doxorubicin, doxorubicin, daunorubicin (vinblastine) daunorubicin, epirubicin, epirubicin, idarubicin, etoposide, teniposide, colchicine, mitoxantrone, dactinomycin, A pharmaceutical composition for inhibiting multi-drug resistance, which is selected from the group consisting of topotecan, trimetrexate, mitramycin, and mitomycin C. The pharmaceutical composition for inhibiting multi-drug resistance of claim 9, wherein the anticancer agent is selected from the group consisting of colon cancer, rectal cancer, bladder cancer, menstrual cancer, breast cancer, and lung cancer.