KR101258537B1 - A novel curcumin derivative with improved water solubility and stability compared with curcumin and method thereof - Google Patents

Abstract

The present invention relates to a new curcumin derivative developed to improve the water solubility and stability that has been pointed out as a problem of curcumin, and more particularly, to a new curcumin derivative having improved water solubility and stability having anticancer activity and a method of preparing the same.

Description

A novel curcumin derivative with improved water solubility and stability compared with curcumin and method

The present invention relates to a new curcumin derivative developed to improve the water solubility and stability that has been pointed out as a problem of curcumin, and more particularly, to a new curcumin derivative having improved water solubility and stability having anticancer activity and a method of preparing the same.

Curcumin is a yellow pigment from the root of a plant called turmeric or Curcuma longa, which has the structure of Chemical Formula 1 below.It has a strong antioxidant effect as well as a spice, and has been used for the treatment of wounds for a long time. It has been used.

[Formula 1]

Recently, many studies on the chemopreventive and anticancer effects have been published. [(A) Bemis, DL; Katz, AE; p Buttyan, R. Expert Opin . Invest. Drugs 2006, 15 , 1191-1200; (b) Nonn, L .; Duong, D .; Peehl, DM Carcinogenesis 2006, 28 , 1188-1196, especially prostate cancer [Deeb, D .; Jiang, H .; Gao, X .; Al-Holou, S .; Danyluk, AL; Dulchavsky, SA; Gautam, SC J. Pharmacol . Exp . Ther . 2007, 321 , 616-625, kidney cancer [Woo, JH; Kim, YH; Choi, YJ; Kim, DG; Lee, KS; Bae, JH; Min, DS; Chang, JS; Jeong, YJ; Lee, YH; Park, JW; Kwon, TK Carcinogenesis 2003, 24 , 1199-1208, liver cancer [Yoysungnoen, P .; Wirachwong, P .; Bhattarakosol, P .; Niimi, H .; Patumraj, S. Clin . Hemorheol . Microcirc. 2006, 34 , 109-115, lymphoma and myeloma [Anuchapreeda, S .; Limtrakul, P .; Thanarattanakorn, P .; Sittipreechacharn, S .; Chanarat, P. Arch . Pharm . Res . 2006, 29 , 80-87, and skin cancer [Odot, J .; Albert, P .; Carlier, A .; Tarpin, M .; Devy, J .; Madoulet, C. Int . J. Cancer 2004, 111 , 381-387] has been reported anti-cancer effect by curcumin.

NF-kB is known as a representative intracellular target protein of curcumin [Tomita, M .; Kawakami, H .; Uchihara, JM; Okudaira, T .; Masuda, M .; TAkasu, N .; Matsuda, T .; Ohta, T .; Tanaka, Y .; Ohshiro, K .; Mori, N. Int . J. Cancer 2006, 118 , 765-772] Curcumin inhibited NF-kB activity by inducing apoptosis and inhibiting enzyme activity such as protein kinase C, epidermal growth factor receptor tyrosine kinase, and HER-2. Results in [(a) Aggarwal, BB; Shishodia, S. Biochem . Pharmacol . 2006, 71 , 1397-1421; (b) Sharma, RA; Gescher, AJ; Steward, W P Eur . J. Cancer 2005, 41 , 1955-1968].

In addition, curcumin has anticancer effects through mitotic spindle disruption and G2 / M phase cell arrest in breast cancer cell line (MCF-7) [Holy, JM Mutat . Res . 2002, 518 , 71-84], colorectal cancer cell line (HT29) exhibits anti-cancer effects through cell shrinkage, DNA degradation and the like [Jiang, MC; Yang-Yen, HF; Yen, JJ; Lin, JK Nutr . Cancer 1996, 26 , 111-120.

The biggest feature of curcumin compared to other anticancer drugs is that it is harmless to the human body. In Phase I, up to 8 g of daily oral dose [Cheng, AL; Hsu, CH; Lin, JK; Hsu, MM; Ho, YF; Shen, TS; Ko, JY; Lin, JT; Lin, BR; Ming-Shiang, W .; Yu, HS; Jee, SH; Chen, GS; Chen, TM; Chen, CA; Lai, MK; Pu, YS; Pan, MH; Wang, YJ; Tsai, CC; H, CY Anticancer Res . 2001, 21 , 2895-2900], and no phase-limiting toxicity was found in Phase II up to 3.6 g of the daily oral dose [Sharma, RA; Euden. A .; Platton, SL; Cooke, DN; Shafayat, A .; Hewitt, HR; Marczylo, TH; Morgan, B .; Hemingway, D .; Plummer, SM; Pirmohamed, M .; Gescher, AJ; Steward, WP Clin. Cancer Res . 2004, 10 , 6847-6854.

However, in spite of such excellent efficacy, curcumin is inevitably restricted to its efficacy in vivo due to its very low water solubility and ultimately causes curcumin's low bioavailability and biodistribution. In fact, when 0.45 to 3.6 g of curcumin was administered to patients with liver metastatic colon cancer, only a very low amount of curcumin or its metabolites were found in serum, but not in the liver at all [Garcea, G. .; Jones, DJ; Sin, R .; Dennison, AR; Farmer, PB; Sharma, RA; Steward, WP; Gescher, AJ; Berry, DP Br . J. Cancer 2004, 90 , 1011-1015.

Curcumin is also unstable under acidic as well as neutral conditions and is known to decompose into various substances such as vanillin, feruroylmethane and ferulic acid [Wang, Y.-J .; Pan, M.-H .; Cheng, A.-L .; Lin, L.-I .; Ho, Y.-S .; Hsido, C.-Y .; Lin, J.-K. J. Pharm . Biomed . Anal. 1997, 15, 1867-1876], curcumin of the α, β-unsaturated 1,3-diketone moiety is because of the nucleophile conjugate addition easily get up as the well-known Michael acceptor [Amolins MW, MW; Peterson, LB; Blagg, BLJ Bioorg . Med . Chem . 2009, 17 , 360-367] based on this, domino Michael addition-retro-Aldol condensation as shown in Figure 1 can be presented as a degradation mechanism. Such instability of curcumin, especially in acidic pH, indicates instability of curcumin in the digestive tract, and thus, it may be apparent that curcumin may act as another factor to reduce the overall bioavailability as well as oral administration efficiency of curcumin.

In conclusion, the low water solubility and instability of curcumin act as a significant limiting factor in curcumin pharmacology.

Recently, water-soluble curcumin has been developed by developing prodrugs (CCMNPEG ³⁵⁰⁰ , CCMNPEG ⁷⁵⁰ ) ([Formula 2]) in the form of conjugates of poly (ethylene glycol) and curcumin having two different lengths (PEG3500 and PEG750). This increase has been reported to increase cytotoxicity (IC ₅₀ ) in several cancer cell lines [Safavy, A .; Raisch, KP; Mantena, S .; Sanford, LL; Sham, SW; Krishna, NR; Bonner, JA J. Med . Chem . 2007, 50 , 6284-6288.

[Formula 2]

On the other hand, curcumin derivatives (Chemical Formula 3) in which 1,3-diketone moiety of curcumin is replaced with electron-rich pyrazole or isoxazole do not act as Michael acceptor, which improves stability and cytotoxicity to cancer cell lines is similar to curcumin. Has been reported [Amolins, MW; Peterson, LB; Blagg, BSJ Bioorg . Med . Chem . 2009 , 17 , 360-367].

(3)

The present invention has been made to solve the above problems, and the object of the present invention is to develop a novel curcumin derivative having improved water solubility and stability compared to curcumin.

It is therefore a main object of the present invention to provide curcumin derivatives with the abovementioned properties.

Another object of the present invention is to provide a method for preparing the curcumin derivative.

Another object of the present invention is to provide an anticancer use of the derivative.

[화학식 4-1]

[화학식 4-2] In order to achieve the above object, the present invention provides a composition comprising a compound of the following [Formula 4-1] and [Formula 4-2].

[Formula 4-1]

[Formula 4-2]

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In the above formula, R ₁ , R ₂ is OH, R ₃ is preferably OCH ₃ but is not limited thereto.

In another aspect, the present invention provides a composition having improved water solubility and stability compared to curcumin containing the compounds of Formulas 4-1 and 4-2, derivatives thereof and salts thereof as an active ingredient.

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In another aspect, the present invention provides a pharmaceutical composition comprising the compound of Formula 4-1 and 4-2 and a pharmaceutically acceptable salt thereof as an active ingredient.
In another aspect, the present invention provides a pharmaceutical composition for preventing or treating cancer, comprising the compounds of Formulas 4-1 and 4-2 and pharmaceutically acceptable salts thereof as an active ingredient.

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In a preferred embodiment of the present invention, the cancer is preferably colon cancer or liver cancer, but is not limited thereto. Cancer pharmaceutical composition for preventing or treating cancer.

In addition, the present invention comprises the steps of a) dissolving 3-chloroacetoacetone in dimethyl sulfoxide (DMSO), and then adding sodium acetate to obtain 3-acetoxyacetylacetone; And

[화학식 4-1]

[화학식 4-2]
상기 화학식에서, R₁, R₂는 OH이고, R₃는 OCH₃인것이 바람직하나 이에 한정되지 아니한다.b) After dissolving boric oxide in dimethylformamide (DMF), tributyl borate and 3-acetoxyacetylacetone obtained from step a) are added and stirred, and 3,4 After adding and stirring dihydroxy-5-methoxybenzaldehyde, and adding and diluting acetic acid diluted in 1,2,3,4-tetrahydroquinoline and dimethylformamide to the following formula 4-1 And 4-2.

[Formula 4-1]

[Formula 4-2]
In the above formula, R ₁ , R ₂ is OH, R ₃ is preferably OCH ₃ but is not limited thereto.

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The compound of the present invention represented by the above formula may be used in the form of a pharmaceutically acceptable salt, and as the salt, an acid addition salt formed by a pharmaceutically acceptable free acid is useful.

The compound represented by the above formula may form a pharmaceutically acceptable acid addition salt according to a conventional method in the art. As the free acid, organic acids and inorganic acids can be used. As the inorganic acids, hydrochloric acid, bromic acid, sulfuric acid, phosphoric acid and the like can be used. As the organic acids, citric acid, acetic acid, lactic acid, tartaric acid, fumaric acid, formic acid, propionic acid, oxalic acid, benzoic acid, gluconic acid, methanesulfonic acid, glycolic acid, succinic acid, 4-toluenesulfonic acid, galacturonic acid, embonic acid, glutamic acid or aspartic acid Can be used.

As used herein, the term "cancer" or "tumor" refers to colorectal cancer, liver cancer, breast cancer, melanoma, squamous cell cancer, colon and generally GI tract, cancer of GIST, lung cancer, head and neck cancer, genitourinary cancer, Hematological cancers, especially leukemia, cervical cancer, uterine cancer, ovarian cancer, testicular cancer, prostate cancer or bladder cancer; Hodgkin's disease or Kaposi's sarcoma, including but not limited to.

The term "anticancer" or "antitumor" as used herein means, but is not limited to, the initiation, progression, or the like, or death of cancer cells, as listed above.

The composition of the present invention can be applied by any route of administration possible for cancer patients such as injections, oral and transdermal agents.

The composition of the present invention can be administered by various methods such as oral administration, parenteral administration, subcutaneous administration, intradermal administration and the like. Preferred dosages of the compounds of the present invention vary depending on the condition and weight of the patient, the severity of the disease, the form of the drug, the route of administration and the duration, and may be appropriately selected by those skilled in the art. Preferably, the compound of the present invention is administered at 0.001-100 mg / kg body weight per day, more preferably 0.01-30 mg / kg body weight. Administration can be done once a day or may be divided several times. The compounds of the present invention in the composition may be present in an amount of 0.0001 to 99% by weight relative to the total weight of the total composition.

The composition of the present invention is generally administered in a pharmaceutical formulation, which is made by mixing it with a pharmaceutically acceptable carrier or diluent. Pharmaceutical preparations include, for example, oral dosage forms such as powders, tablets, capsules, solutions, and the like, transdermal absorbents, lotions, emulsions and suspensions, external preparations such as patches, creams, cataplasmas, intramuscular injections, intramuscular injections, etc. Injections, and the like.

These pharmaceutical compositions are prepared by conventional methods. Particularly preferred is an external preparation, and specific examples thereof include cream, ointment, gel, lotion, emulsion, stick, pack, and organic solvent solution, but the scope of the formulation is limited thereto. It is not there.

Pharmaceutically acceptable carriers or diluents can be any conventional ones used in the pharmaceutical art and do not react with the compounds of the present invention. Suitable examples of pharmaceutically acceptable carriers or diluents for powders, tablets, capsules and the like are excipients such as corn starch, lactose, mannitol, microcrystalline cellulose, disintegrants, tablets such as croscarmellose sodium, potato starch, white sugar, tablets Binders such as gelatin, gum arabic, methylcellulose, ethylcellulose, povidone and the like, and lubricants such as magnesium stearate, hard silicic anhydride and talc.

Tablets may be coated by conventional methods with a coating agent such as hydroxypropylmethylcellulose, hydroxypropylmethylcellulose phthalate, cellulose acetate phthalate, titanium oxide, polysorbate, purified white sugar and the like.

Examples of ingredients used in external preparations for skin include liquid oils, oils and fats, beeswax, hydrocarbons, higher fatty acids, higher alcohols, esters, surfactants, humectants, water-soluble high molecular compounds, thickeners, coatings, lower alcohols, and polyhydric alcohols. , Sugars, amino acids, organic amines, pH adjusters, antioxidants, fragrances, water and the like can be appropriately blended as necessary. You may use 1 or more types of these components, respectively.

Suitable examples of the adhesive base of the patch include an acrylic copolymer, a polymer base such as polyvinylpyrrolidone, polyisobutylene, and the like, a plasticizer such as triethyl citrate, triethylacetyl citrate, glycerin, propylene glycol, and polyethylene glycol. Can be mentioned.

Injectables can be prepared by dissolving a salt of 3-hydroxyflavone in distilled water for injection, but if necessary, isotonic agents, analgesics, pH adjusters, dissolution aids, buffers, preservatives and the like can be added. Injectables can also be in the form of suspensions, which can be prepared by suspending the compound of the present invention in distilled water or vegetable oil for injection, as necessary, adding a base, a suspending agent and the like. Injectables may also be in the form of powders or lyophilized, which may dissolve upon use and further add excipients and the like.

The above formulations may be provided as controlled release agents by appropriate methods such that a certain amount of drug can be released at a constant rate and absorbed into the patient's circulatory system according to the patient's dosing schedule. Examples of controlled release include tablets in the form of a matrix or in the form of a coating film, granules or capsules encapsulated therein, or transdermal absorbents.

Pharmaceutical dosage forms of the compositions of the present invention may be used alone or in combination with other pharmaceutically active compounds, as well as in a suitable collection.

In various embodiments of the invention, the anticancer agent that can be used together is a chemotherapeutic agent, radiotherapy agent, or both.

Hereinafter, the present invention will be described.

In the present invention, 1,3-diketo moiety of curcumin is expected by increasing the electron density in the central skeleton of the curcumin derivative, 1) increase in water solubility due to the enhancement of polarity and 2) curcumin degradation by Michael addition of nucleophiles An electron donating group was introduced into the central methylene unit located at. As an electron donor, an acetoxy group, which is relatively easy to synthesize, was used, and both aromatic rings were synthesized by adding a derivative ([Chemical Formula 4]) having one more OH group than curcumin to enhance water solubility. By evaluating and the like, the usefulness of the new curcumin derivative is demonstrated and the present invention has been completed.

[Formula 4]

Hereinafter, the present invention will be described in detail.

The present invention synthesized a curcumin derivative represented by the formula (4). Specifically, Figure 2 shows the process of synthesizing curcumin derivatives from commercially available starting materials. 3-acetoxyacetoacetone (2), the main intermediate required for the synthesis of curcumin derivatives, was synthesized by the acetylation reaction of 3-chloroacetoacetate (1), and the synthesized compound 2 was known in [Lin, L .; Shi, Q .; Su, C.-Y .; Shih, CC-Y .; Lee, K.-H. Bioorg . Med . Chem . 2006, 14 , 2527-2534] complexed with boric anhydride and converted to the desired curcumin derivatives through 3,4-dihydroxy-5-methoxybenzaldehyde and 1,5-bis-aldol condensation.

Hereinafter, the results of analyzing the water solubility, stability and anticancer activity of the synthesized curcumin derivative will be described in detail.

As a result of measuring the water solubility of the synthesized curcumin derivative is shown in FIG. Since the vertical axis (Relative Nephelometry Unit, RNU) refers to the scattering of light by undissolved molecules, the smaller the scattering degree, the higher the solubility. As shown in FIG. 3, the solubility of curcumin is significantly low even at low concentrations (5 mM), and it is confirmed that the solubility decreases rapidly as the concentration increases, but the solubility of the curcumin derivative is consistently high regardless of the concentration.

In order to compare the stability of the synthesized curcumin derivatives and curcumin cultured in the cell culture (37 ℃) is shown in Figures 4 and 5 below. 4 shows the results of curcumin culture, 4a is the chromatogram of curcumin before incubation ( t = 0 min) and 4b is the chromatogram after 30 minutes ( t = 30 min) in culture, halving the amount of curcumin remaining after 30 minutes in culture. Decreasing shows that the curcumin's half-life ( t _1/2 ) is about 30 minutes. In FIG. 5, which shows the culture results of curcumin derivatives, FIG. 5A is a chromatogram of curcumin derivatives before incubation ( t = 0 min), and FIG. 5B is a chromatogram after twelve hours of culture ( t = 1 2 hours). show that remains stable without decomposition even after incubation _{(t 1/2> 12 h} ).

Meanwhile, as a result of anticancer activity analysis (FIG. 6), curcumin showed good anticancer activity in colorectal cancer cell line (HCT116) and breast cancer cell line (MCF-7), whereas curcumin derivatives were shown in colorectal cancer cell line (HCT116) and liver cancer cell line (Huh-). 7) showed similar anti-cancer activity to curcumin or superior to curcumin.

As can be seen from the above-mentioned problem solving means and the following examples, in order to overcome the low water solubility and instability of curcumin, the electron donor acetoxy group was introduced into the central methylene unit of curcumin and a hydroxyl group was added to both aromatic rings. Curcumin derivatives have been developed. The new curcumin derivatives can overcome the low bioavailability that has been pointed out as a curcumin's shortcoming by confirming its cytotoxicity against cancer cell lines comparable to curcumin but showing dramatically increased water solubility and stability in the extracellular environment compared to curcumin. It is expected to be.

1 shows a process in which curcumin is degraded and lost its utility in an extracellular environment.
Figure 2 shows the process of synthesis of the new curcumin derivatives.
Figure 3 shows the results of water solubility verification of curcumin and novel curcumin derivatives.
Figure 4 shows the stability of curcumin in cell-free medium. 4a is the chromatogram of curcumin before incubation ( t = 0 min) and 4b is the chromatogram after 30 minutes in culture ( t = 30 min).
Figure 5 shows the stability of the new curcumin derivative in cell-free medium. Figure 5a shows the chromatogram of the curcumin derivative before incubation ( t = 0 min) and Figure 5b shows the chromatogram after twelve hours in culture ( t = 1 2 hours). It is a graph.
Figure 6 shows the anticancer activity of curcumin and novel curcumin derivatives in colorectal cancer (HCT116), liver cancer (Huh-7), breast cancer (MCF-7) cell lines.
7 is a chemical formula showing the structure of a novel curcumin derivative of the present invention.

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for illustrating the present invention, it will be apparent to those skilled in the art that the scope of the present invention is not to be construed as limited by these examples.

All reagents and solvents including 3-chloroacetoacetate, sodium acetate (NaOCOCH ₃ ) and MTT [(3- (4,5-Dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide)] used in the present invention are sigma- It was purchased from Aldrich.

In addition, DMEM (Dulbeccos Modified Eagle Medium), penicillin, streptomycin and calf serum (FBS) were purchased from Invitrogen.

Example 1: Preparation of 3- acetoxyacetylacetone (2)

3-chloroacetoacetone (1, 4.4 mmol) is dissolved in DMSO (5 ml), then sodium acetate (8.8 mmol) is added and stirred at room temperature for 3 hours. Water (20 mL) is added to the reaction and extracted three times with diethyl ether (30 mL). Magnesium sulfate is added to the obtained organic layer to remove water, and then filtered through a filter paper. After concentration under reduced pressure, the obtained concentrate was purified by silica gel column chromatography (hexane: ethyl acetate = 8: 1) to give 3-acetoxyacetylacetone (2) in the form of an oil (yield 60%).

Example 2: Preparation of Curcumin Derivatives

Boric oxide (3.2 mmol) was dissolved in DMF (1 mL), tributyl borate (6.4 mmol) and 3-acetoxyacetylacetone (2, 3.2 mmol) obtained above were added thereto, and the mixture was stirred under reflux at 65 ° C. for 20 minutes. . 3,4-dihydroxy-5-methoxybenzaldehyde (6.4 mmol) was added to the mixture, and the mixture was stirred under reflux at 65 ° C for 10 minutes. Then, 1,2,3,4-tetrahydroquinoline (0.1 mL) and DMF (1 mL) were added. Dilute acetic acid (0.3 mL) was added and stirred under reflux again at 95 ° C. for 4 hours. After cooling the reaction to 15 ° C., 20% acetic acid was added and stirred under reflux at 70 ° C. for 1 hour. The reaction mass was cooled to 15 ° C. again, magnesium sulfate was added to the organic layer obtained by extracting three times with ethyl acetate (30 mL), and water was removed. After concentration under reduced pressure, the obtained concentrate was purified by silica gel column chromatography (hexane: ethyl acetate = 1: 1) to obtain a curcumin derivative as a mixture of diketo form and keto-enol form (green powder, yield 35%, di-). keto: keto-enol = 1.25: 1); For di-keto form, ¹ H NMR (500 MHz, CDCl ₃ ) δ (ppm) 7.64 (d, J = 19.5 Hz, 2H), 6.90 (br s, 2H), 6.86 (d, J = 19.5 Hz, 2H ), 6.70 (br s, 2 H), 5.73 (s, 1 H), 3.96 (s, 6 H), 2.12 (s, 3 H); For keto-enol form, ¹ H NMR (500 MHz, CDCl ₃ ) δ (ppm) 15.49 (s, OH), 7.51 (d, J = 19.5 Hz, 2H), 6.87 (br s, 2H), 6.60 (br s, 2H), 6.46 (d, J = 19.5 Hz, 2H), 3.91 (s, 6H), 2.12 (s, 3H); LC / MS (ESI) m / z Found: 459.55 [M + H] ⁺ ; Calcd for C ₂₃ H ₂₂ O ₁₀ : 458.12.

Example 3: Determination of Water Solubility of Curcumin Derivatives

Dissolve the synthesized curcumin derivative in DMSO to obtain a 250, 200, 150, 50, 25, 10 mM stock solution. Dilute each stock solution to 2500, 2000, 1500, 1000, 500, 250, 100 μM by adding PBS buffer at the rate of 1% DMSO-99% PBS buffer (pH 7.4). Take 250 μL of each stock solution into a 96-well microplate and measure forward light scattering using a NEPHELOstar laser based microplate nephelometer. Solubility is calculated from forward light scattering using BMG LABTECH NEPHELOstar Galaxy Evaluation software.

Example 4 Determination of Chemical Stability of Curcumin in Cell Cultures

Take 2 μl of stock solution (10 mM) of the synthesized curcumin derivatives dissolved in methanol, add to RPMI 1640 medium (198 μl), and incubate at 37 ° C. 0, 5, 10, 15, 30 minutes after the incubation, take 100 μl of the aliquots, respectively, and filter the filtrate. HPLC analysis conditions: C-18 reverse phase column; flow rate, 1 mL / min; detection, UV 300 nm; mobile phase, 20 min (60% acetonitrile and 0.1% acetic acid)

Example 5 Determination of Chemical Stability of Curcumin Derivatives in Cell Culture Solution

Take 2 μl of stock solution (10 mM) of the synthesized curcumin derivatives dissolved in methanol, add to RPMI 1640 medium (198 μl), and incubate at 37 ° C. 0, 30, 60 minutes, and 12 hours after incubation, take 100 μl of the aliquot and filter the filtrate, respectively, and analyze the filtrate via HPLC. HPLC analysis conditions: C-18 reverse phase column; flow rate, 1 mL / min; detection, UV 300 nm; mobile phase, 0-4 min (20% acetonitrile and 0.1% acetic acid), 5-9 min (25% acetonitrile and 0.1% acetic acid), 10-15 min (35% acetonitrile and 0.1% acetic acid), 16- 20 min (80% acetonitrile and 0.1% acetic acid), 21-25 min (20% acetonitrile and 0.1% acetic acid).

Example 6: Determination of anticancer activity of curcumin derivatives :

Cell viability was performed for the liver cancer (Huh-7), colon cancer (HCT116), and breast cancer (MCF-7) cell lines, respectively, and HS27, a normal cell line, was set as a control. Count the cell suspension using a haemacytometer and add 180 μL of the cell suspension of the desired concentration to each well of a 96 well plate [Blank: 180 μL of medium and 20 μL of PBS, Control: 180 μL of cell suspension and 20 μL of PBS ]. After dissolving curcumin derivative in phosphate buffer solution, add 20 μL of each well to each well and incubate the 96-well plate for 72 hours at 37 ^o C, 5% CO ₂ , 95% relative humidity. Remove the sample, add 100 μL of MTT solution to each well, incubate for 4 hours, and carefully remove the MTT diluent. Add 100 μL of DMSO to each well, stir with a plate shaker for 15-20 minutes, and measure the absorbance at wavelength 540 nm using an ELISA reader. This absorbance represents the amount of MTT reduced by cells and is present in each well. Is proportional to the number of viable cells.

Claims (6)

A composition comprising the compound of [Formula 4-1] and [Formula 4-2].

[Formula 4-1]

[Formula 4-2]
In the above formula, R ₁ and R ₂ is OH, R ₃ is OCH ₃ A composition having improved water solubility and stability compared to curcumin comprising the compounds of the following [Formula 4-1] and [Formula 4-2], or salts thereof as an active ingredient.

[Formula 4-1]

[Formula 4-2]
In the above formula, R ₁ and R ₂ is OH, R ₃ is OCH ₃ A pharmaceutical composition comprising the compounds of [Formula 4-1] and [Formula 4-2], and a pharmaceutically acceptable salt thereof as an active ingredient.

[Formula 4-1]

[Formula 4-2]
In the above formula, R ₁ and R ₂ is OH, R ₃ is OCH ₃ A pharmaceutical composition for preventing or treating cancer comprising the compounds of the following [Formula 4-1] and [Formula 4-2], and pharmaceutically acceptable salts thereof as an active ingredient.

[Formula 4-1]

[Formula 4-2]
In the above formula, R ₁ and R ₂ is OH, R ₃ is OCH ₃ The pharmaceutical composition for preventing or treating cancer according to claim 4, wherein the cancer is colorectal cancer or liver cancer. a) dissolving 3-chloroacetoacetone in dimethyl sulfoxide (DMSO) and then adding sodium acetate to obtain 3-acetoxyacetylacetone; And
b) After dissolving boric oxide in dimethylformamide (DMF), tributyl borate and 3-acetoxyacetylacetone obtained from step a) are added and stirred, and 3, 4-dihydroxy-5-methoxybenzaldehyde was added and stirred, followed by addition and stirring of 1,2,3,4-tetrahydroquinoline and acetic acid diluted in dimethylformamide. Process for the preparation of compounds of 1 and 4-2.

[Formula 4-1]

[Formula 4-2]
In the above formula, R ₁ and R ₂ is OH, R ₃ is OCH ₃

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