KR100933770B1 - Secoisorariciresin derivatives and anticancer agents using the same - Google Patents

Abstract

The present invention is a novel structure of secoisolariciresinol derivatives having a strong cytotoxicity against cancer cells and the effect of inhibiting the polymerization of intracellular actin cytoskeleton and the compound as an active ingredient. It relates to anticancer drugs.

Description

Secoisorariciresinol derivatives and anticancer agents having the same as an active ingredient {Derivatives of secoisolariciresinol, and anticancer agent containing the same as active ingredient}

The present invention is a novel structure of secoisolariciresinol derivatives having a strong cytotoxicity against cancer cells and the effect of inhibiting the polymerization of intracellular actin cytoskeleton and the compound as an active ingredient. It relates to anticancer drugs.

Actin is a major component of cytoskeleton and plays an important role in the areas of cell growth, division, signaling, cell-cell aggregation and wound healing. Actin-related proteins are involved in the process of polymerizing actin monomers into actin filaments. As the actin filaments stick or fall, the cells change shape, cell division, and cell bonding occur. Actin and its related proteins, which play this role, have become the main target for the development of cancer therapeutics [Jordan, MA; Wilsont, L. Curr. Opin. Cell Biol . 1998, 10 , 123-130; Giganti, A .; Friederich, E. Prog. Cell Cycle Res . 2003, 5 , 511-525. Following this trend, many cytotoxic natural products interacting with actin have been reported recently. Examples include latrunculin A, jasplakinolide, swingholide A, pectenotoxins and the like [Allingham, SVA; Raymen, I. Cell Mol. Life Science . 2006, 63 , 2119-2134; Fenteany, G .; S. Curr. Top. Med. Chem . 2003, 3 , 593-616; Spector, I .; Braet, F .; Shochet, NR; Bubb, MR Microsc. Res. Tech . 1999, 47 , 18-37.

The well-known medicinal herb, Trichosanthes kirilowii , is a perennial vine and grows on hills and forest edges in southern and central Korea. In oriental medicine, the roots of the sky feathers are called 'fruit tree roots', 'bushing roots' or 'cheonhwahwa', and the fruit of the sky feathers is called 'fruits' and the seed of the sky feathers is called 'bapluin'. In addition, the herbal medicine and root of the sky is mainly used as a medicine such as bruises, blood, jaundice, skin disease, and seeds are mainly used as expectorant, Jinhae, analgesic, anti-inflammatory agent. 4 books; Seoul Press, 1996; p 172]. As such, the fruit and root of the sky has been used in folklore for a long time, but the report on the composition and physiological activity of each part is still insufficient.

On the other hand, the present inventors have reported to separate the flavone-based material from the sky fruit (earth fruit) based on the folk medicinal effect that Trichosanthes kirilowii has anti-cancer efficacy [Rahman, Md. Aziz Abdur and Moon, Surk-Sik. Bull. Korean Chem. Soc . 2007, 28 (8), 1261-1264]

The present invention extracts and fractions the skyseed seeds (Gluluin) with an organic solvent to obtain a new structure of secoisolariciresinol (secoisolariciresinol) derivatives, the new derivatives are human lung cancer cell line A549, melanoma cell line Sk-Me-2 The present invention was confirmed by showing strong cytotoxicity against, and confirming the efficacy of inhibiting the polymerization of actin cytoskeleton in keratinocyte line HaCaT cells, which is effective as an anticancer agent.

It is an object of the present invention to provide a secoisolariciresinol derivative of novel structure.

Another object of the present invention is to provide a medicinal use of the novel secoisorariciresinol derivatives described above as an anticancer agent.

Another object of the present invention is to isolate hanultarin ((-)-(2R, 3R) -1- O- feruloylsecoisolariciresinol), a compound of the above-mentioned novel secoisorariciresinol derivatives, from the skyseed seeds (Juluin). To provide a way.

In order to achieve the above object, the present invention is characterized by the Seiko isolari resirinol derivative represented by the following formula (1) or a pharmaceutically acceptable salt thereof.

In Formula 1, R ₁ , R ₂ , and R ₃ each represent a hydrogen atom or an alkyl group having 1 to 6 carbon atoms.

In addition, the secoisorariciresinol derivative represented by the formula (1) exhibits excellent cancer cytotoxicity and the inhibitory effect of polymerization of actin, a cytoskeleton component of the cell, and the present invention is the secoisolarishsirishi represented by the formula (1). It is characterized by an anticancer agent containing a sol derivative as an active ingredient. The anticancer treatment by the use of the secoisorariciresinol derivative represented by the formula (1), the method of treatment by killing cancer cells and the treatment method involving the disappearance of cancer cells obtained by inhibiting the polymerization of actin in the growth of cancer cells Say.

In addition, the present invention in the secoisorariciresinol derivative represented by the formula (1), wherein R ₁ , R ₂ , and R ₃ are each a methyl group, aka tarin (hanultarin) is a solvent extraction from the sky seeds, silica gel column It is characterized by a method of separation and obtaining by chromatography and C18 reversed phase HPLC.

The secoisorariciresinol derivative represented by Chemical Formula 1 according to the present invention may be used in the form of a pharmaceutically acceptable salt, and acid salts formed by pharmaceutically acceptable free acid are useful as salts. Do. Inorganic acids and organic acids can be used as the free acid. Hydrochloric acid, bromic acid, sulfuric acid, phosphoric acid, etc. may be used as the inorganic acid, and citric acid, acetic acid, lactic acid, tartaric acid, maleic acid, fumaric acid, gluconic acid, methanesulfonic acid, glyconic acid, succinic acid, 4-toluene Sulfonic acid, gluturonic acid, embonic acid, glutamic acid, aspartic acid and the like can be used.

In the secoisorarishrecinol derivative represented by the formula (1), the substituents R ₁ , R ₂ , and R ₃ each represent a hydrogen atom or an alkyl group having 1 to 6 carbon atoms. Preferred as the secoisolaryrrishinol derivative represented by the formula (1), substituents R ₁ , R ₂ , and R ₃ are each alkyl selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, and isobutyl. Compound. Particularly preferred secoisolarishilsinol derivatives represented by the above formula (1) are compounds in which the substituents R ₁ , R ₂ , and R ₃ are each methyl. In the present invention, the substituents R ₁ , R ₂ , and R ₃ , each of which is a methyl group, are separated from the seed of Secoisolarysirinol derivatives represented by Formula 1 and named as 'hanultarin'. Therefore, 'hanultarin' can be obtained as a natural product among the secoisorarishrecinol derivatives according to the present invention.

In addition, the secoisolarysirinol derivative represented by Formula 1 according to the present invention can be easily synthesized by a known organic synthesis method if a person skilled in the art of organic synthesis. For example, 1-halo-2,3-bis (3,4-dihydroxybenzyl) butan-4-ol and Peruroic acid (feruloic acid) are used as raw materials for the coupling and alkyl substitution reactions. It can be carried out to synthesize the secoisorariciresinol derivative represented by the formula (1). In addition, the modification of the raw material used suitably for the target compound is also obvious at the supplier level.

The present invention also includes a method of separating the sky tarin (hanultarin) as a natural product from the sky seed as a right bar, if more specifically describe the separation method for accommodating sky tarin (hanultarin) as a natural product in more detail as follows.

First, finely crushed crushed pulverized seed (goluin) is extracted with an organic solvent. The organic solvent used for the extraction is selected from the group consisting of aliphatic alcohols having 1 to 6 carbon atoms, ethyl acetate and hexane including methanol, ethanol, n-propanol, isopropanol, n-butanol, isobutanol, pentanol, and hexanol Single or mixed solvents may be used. The alcohol used for extraction may use anhydrous alcohol or a water soluble alcohol containing water in the range of 1 to 80% by volume, preferably 30 to 60% by volume. Extraction using the above organic solvent may be repeated once to several tens of times, and the extract obtained by extracting is collected and concentrated under reduced pressure to obtain a brown viscous liquid.

Then, the solvent extract obtained above is dissolved in an alcohol having 1 to 6 carbon atoms, extracted with hexane, and the alcohol layer is obtained and concentrated. The alcohol used to dissolve the extract may use anhydrous alcohol or a water soluble alcohol containing water in the range of 1 to 80% by volume, preferably 30 to 60% by volume. Extracting the alcohol solution with hexane may also be repeated once to several dozen times. The alcoholic solution layer is separated and concentrated under reduced pressure to give a brown liquid.

Then, the concentrate obtained above is subjected to silica gel column chromatography to obtain an active fraction. For example, primary silica gel column chromatography is performed by increasing the polarity of the mixture of methylene chloride, methanol and water step by step as the eluent. In primary silica gel column chromatography, it can be carried out with increasing polarity within the range of 100: 0: 0 to 90: 9.5: 0.5 in a mixed volume ratio of methylene chloride / methanol / water as eluent. Then, secondary silica gel column chromatography is performed with increasing polarity step by step using a mixture of hexane, ethyl acetate and methanol as eluent. In secondary silica gel column chromatography, a mixed volume ratio of hexane / ethyl acetate as the eluent may be performed while increasing the polarity within the range of 8: 2 to 1: 9.

Then, the active fractions obtained by silica gel column chromatography were subjected to C18 reversed phase high pressure liquid chromatography (HPLC) to obtain a secoisorariciresinol derivative represented by Chemical Formula 1 wherein R ₁ = R ₂ = R ₃ = methyl. As the eluate, water-soluble acetonitrile was used. Specifically, water-soluble acetonitrile or water-soluble alcohol containing 1 to 70% by volume of water, preferably 30 to 60% by volume, may be used.

For the structural analysis of the active material obtained by performing a series of extraction and fractionation process as described above, mass spectrometry, infrared spectroscopy, ultraviolet spectroscopy, hydrogen nuclear magnetic resonance analysis, carbon nuclear magnetic resonance analysis were performed. More detailed analysis will be described in Example 2 below.

On the other hand, the present invention is a pharmaceutical composition comprising a secoisorariciresinol derivative represented by the formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient, including a pharmaceutically acceptable carrier, excipient, diluent Include as a scope of rights. Since the pharmaceutical composition is used as an anticancer agent to exhibit an effective drug, the present invention also includes a pharmaceutical use of the pharmaceutical composition as an anticancer agent is also included in the scope of the present invention.

Carriers, excipients and diluents included in the pharmaceutical composition of the present invention are commonly used in the pharmaceutical field, for example, lactose, dextrose, sucrose, mannitol, starch, acacia gum, alginic acid salt, gelatin, phosphoric acid Calcium salt, cellulose, methylhydroxy benzoate, magnesium stearate, etc. are mentioned. The formulation may further comprise fillers, anti-coagulants, lubricants, wetting agents, emulsifiers, preservatives and the like. In addition, the anticancer agent of the present invention may be formulated using methods known in the art so that the active ingredient can provide rapid release, slow release or delayed release after administration to a mammal.

In addition, the pharmaceutical compositions of the present invention may be formulated as a conventional formulation in the pharmaceutical art and administered via various routes including oral, transdermal, subcutaneous, intravenous or intramuscular. For example, the compounds according to the invention may be in the form of tablets containing starch or lactose, in the form of capsules alone or in the form of excipients, or in the form of elixirs or suspending agents containing chemicals which flavor or color. It can be administered orally, orally or sublingually. Liquid formulations may be suspending agents (eg, methyl cellulose, semisynthetic glycerides such as witepsol or mixtures of apricot kernel oil with PEG-6 esters or PEG-8 with caprylic / capric glycerol). Pharmaceutically acceptable additives such as glyceride mixtures such as mixtures of lides). In addition, parenteral preparations can be injected, for example, intravenously, intracavernosally, intramuscularly, subcutaneously and intratracheally. For parenteral administration, it is most preferred to use it in the form of a sterile aqueous solution, in which the solution contains other substances (e.g. salts or monosaccharides such as mannitol, glucose) to have isotonicity with the blood. You may.

In addition, the dosage of the compound represented by Formula 1 to the human body may vary depending on the age, weight, sex, dosage form, health condition and degree of disease of the patient, when the weight is 70 kg based on an adult patient Generally, it is 0.01 to 400 mg / day, and may be dividedly administered once to several times a day at regular time intervals according to the judgment of a doctor or a pharmacist.

The present invention as described above will be described in more detail based on the following examples, but the present invention is not limited thereto.

[ Example ]

Example 1 Isolation of Hanultarin from Gluluin

10 Kg of the fly seed (Jwaruin) was crushed and soaked in 80% water-soluble methanol (20 L) and placed for 1 week at room temperature. The water-soluble methanol portion was decanted off, and the remaining residue was taken up in the same way with ethyl acetate (18 L) and ethyl acetate-hexane (6: 1, 14 L). All of the pools were combined and concentrated under reduced pressure to give a brown viscous liquid (2.6 Kg). This liquid was dissolved in 30% aqueous methanol (1.4 L) and then extracted 10 times with hexane (1.2 L). The remaining water soluble methanol portion was concentrated to give a brown liquid portion (101 g). This was again dissolved in water and methylene chloride (1: 1, 2.4 L) as much as possible, and methylene chloride portion (30 g) was silica gel column chromatography (silica gel 60, 70-230 mesh, Merck, 50 id × 220 mm). . A mixture of methylene chloride, methanol and water was used as the eluent to increase the polarity step by step (variing the volume ratio from 100: 0: 0 to 90: 9.5: 0.5 v / v / v) to obtain a total of 14 fractions. Fraction 4 (6.9 g, R _f = 0.4 to 0.6, Eluent: methylene chloride / methanol / water, 90: 9.5: 0.5 v / v / v) showed a silica gel column chromatography (eluent: hexane / ethyl). Acetate = 8/2 → 1/9 v / v) to give 8 fractions. From this fraction 7 (2.9 g, R _f = 0.5 to 0.6, eluent: methylene chloride / methanol / water = 90: 9.5: 0.5 v / v / v) was again subjected to C18 reverse phase medium pressure liquid chromatography (30-100% aqueous methanol). , 220 min, 8 mL / min) and divided into 8 fractions.

Among them, fraction 3 (254 mg, R _f = 0.5 to 0.6, eluent: methylene chloride / methanol / water, 90: 9.5: 0.5 v / v / v) was purified by C18 reverse phase HPLC (Senshu pak, Pegasil ODS, 20 id x 250). mm, 30-40% water soluble acetonitrile, 75 min, 7 mL / min, UV 225 nm) to give pinoresinol (18.3 mg, R _t 31.2 min) and 4-ketopinoresinol (4-ketopinoresinol, 6.32). mg, R _t 36.6 min).

And fraction 4 (354 mg, R _f = 0.5-0.6, eluent: methylenechloride / methanol / water, 90: 9.5: 0.5 v / v / v) was purified by C18 HPLC (30-60% aqueous acetonitrile, 60 min, 7 mL / min, UV 225 nm) gave secoisolariciresinol (secoisolariciresinol, 8.5 mg, R _t 25.2 min) and light tan viscous liquid (hanultarin, 28 mg, R _t 37.2 min).

Fraction 5 (350 mg, R _f = 0.5-0.6, eluent: methylenechloride / methanol / water, 90: 9.5: 0.5 v / v / v) was purified by HPLC (25-70% aqueous acetonitrile, 50 min, 7 mL). / min, UV 225 nm) to give 2,3-diferroyl secoisolariciresinol (2,3-diferuloylsecoisolariciresinol, 40 mg, R _t 33.6 min).

Example 2 Characterization of the Chemical Structure of Hanultarin

Spectroscopic analysis was performed to identify the chemical structure of the empty tarin (hanultarin) obtained by the separation method of Example 1. 1 to 4 show mass spectrometry, infrared (IR) spectroscopy, hydrogen nuclear magnetic resonance ( ¹ H NMR) spectra, and carbon nuclear magnetic resonance ( ¹³ C NMR) spectra of an empty tarin (hanultarin), respectively. Attached.

According to the mass spectrometry, the mass of hanultarin was measured as 538 (HRTOFMS [M + H] ⁺ : m / z 539.2294; Theoretical value, 539.2281). From this the molecular formula is C ₃₀ H ₃₄ O ₉ . Ultraviolet (UV) spectral spectra showed that maximum absorption values in methanol were observed at 326, 288, and 204 nm. The characteristic peaks shown in the infrared (IR) spectral spectrum are as follows: hydroxy group (3409 cm ^-1 ), ester (1702 cm ^-1 ), double bond (1637 cm ^-1 ), and aromatic ring (1602, 1513 cm ^-1 ). Two double-bonded hydrogens [δ 7.56 (1H, J = 16 Hz) and 6.35 (1H, J = 16 Hz)], aromatic hydrogens [δ 7.05 (1H, dd, J ) from ¹ H NMR and ¹³ C NMR spectra = 8.4, 1.6 Hz), 6.81 (1H, d, J = 8.4 Hz), and 7.17 (1H, d, J = 1.6 Hz)] and methoxy hydrogen [δ 3.88 (3H, s)].

From the above data, it can be seen that part of the chemical structure of the hanultarin contains a feruloyl (4-hydroxy-3-methoxyphenylcinnamoy) group. Several pairs of aromatic cyclic hydrogens [δ 6.55 (1H, dd, J = 7.6, 1.6 Hz) and 6.664 (1H, d, J = 7.6 Hz); 6.62 (1H, d, J = 1.6 Hz) and 6.54 (1H, dd, J = 8.0, 2.0 Hz), 6.662 (1H, d, J = 8.0 Hz) and 6.58 (1H, d, J = 1.6 Hz)] In view of the presence of two methoxy hydrogens [δ 3.71 (3H, s) and 3.7 (3H, s)], two 4-hydroxy-3-methoxyphenyl groups were found. In addition, methylene hydrogen [δ 2.71 (2H, dd, J = 14.0, 7.2 Hz) and 2.61 (2H, dd, J = 13.2, 8.0 Hz)], two methine hydrogens [δ 2.24 (m), and 1.97 ( m)], two pairs of methylene hydrogen [δ 4.33 (1H, dd, J = 11.2, 6.0 Hz), 4.10 (1H, dd, J = 11.2, 6.4 Hz), 3.53 (1H, dd, J = 11.2, 6.8 Hz) and 3.67 (1H, dd, J = 11.2, 6.0 Hz)]. From the analysis of these data, hanultarin is composed of two 1,3,4-trisubstituted aromatic rings, -CH ₂ -CH (CH ₂ O-)-CH (CH ₂ O-)-CH _2- . I found out to include. These substructures led to the interpretation of the HMBC spectrum.

In summary, the chemical structure of the empty tarin (hanultarin) isolated in Example 1 is represented by the following formula (1a), and the compound name is 1- o -ferroyl secoisorariciresinol (1- o). feruloylsecoisolariciresinol). The optical activity of isolated hanultarin was -19.7 (c 0.36, methanol), and it was previously known analogous material, (-)-(2R, 3R) -secoisorarishrecinol ((-)-(2R, 3R). )-secoisolariciresinol) has a value of (-), so the stereostructure of the sky carbon is considered to be the same.

Spectroscopic analytical data of hanultarin measured in Example 2 are as follows: pale brown viscous liquid; Optical activity -19.7 ° (c 0.36, methanol); HRTOFMS [M + H] + m / z: 539.2294 (calculated: 539.2281), molecular formula C ₃₀ H ₃₄ O ₉ + H); Ultraviolet spectrum (ν _max , MeOH) 326, 288, 204; Infrared spectrum (KBr): maximum absorption frequency 3409, 2950, 1702, 1637, 1602, 1513 1467, 1429, 1387, 1244, 1210, 1157, 1023 cm ⁻¹ ; ¹ H NMR and ¹³ C NMR (CD ₃ OD) see Table 1 below.

[Shoemaking]

On the other hand, the novel compound represented by Formula 1 according to the present invention can be formulated in various forms according to the purpose. The following illustrates some 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.

Formulation 1: tablet (direct pressure)

After sifting 5.0 mg of active ingredient, 14.1 mg of lactose, 0.8 mg of crospovidone USNF, and 0.1 mg of magnesium stearate were mixed and pressed to form a tablet.

Formulation 2: Tablet (Wet Granulation)

After sifting 5.0 mg of the active ingredient, 16.0 mg of lactose and 4.0 mg of starch were mixed. 0.3 mg of polysorbate 80 was dissolved in pure water and then an appropriate amount of this solution was added and then atomized. After drying, the fine particles were sieved and mixed with 2.7 mg of colloidal silicon dioxide and 2.0 mg of magnesium stearate. The granules were pressed into tablets.

Formulation 3: Powders and Capsules

5.0 mg of the active ingredient was sieved, followed by mixing with 14.8 mg of lactose, 10.0 mg of polyvinyl pyrrolidone, and 0.2 mg of magnesium stearate. No. solid the mixture using a suitable device. Filled in 5 gelatin capsules.

Formulation 4: Injection

Injectables were prepared by containing 100 mg as the active ingredient, followed by 180 mg of mannitol, 26 mg of Na ₂ HPO ₄ .12H ₂ O and 2974 mg of distilled water.

Experimental Example

Experimental Example 1. Measurement of cancer cytotoxicity of sky tarin

The cancer cytotoxicity of the sky tarin cancer cell lines A549, SK-Mel-2 and B16F1 isolated from the present invention was measured by SRB (colorimetric sulforhodamine B) method [Skehan, P .; Storeng, R .; Scudiero, D .; Monks, A .; McMahon, J .; Vistica, D .; Warren, JT; Bokesch, H .; Kenney, S .; Boyd, MR J. Natl . Cancer Inst . 1990, 82 , 1107; Rahman, Md. Aziz Abdur, Cho, Soon-Chang, Song, Junsik, Mun, Hyeong-Tae, Moon, Surk-Sik. Planta Medica , 2007, 73 , 1089-1094].

The cell density of each cell line was 1 × 10 ⁵ , 1 × 10 ⁵ , 2 for human lung cancer cell line A-549, human color tumor cancer cell line SK-MEL-2, and mouse black tumor cancer cell line B16-F1, respectively. 10 ⁴ cells / mL were used. 100 mL of the medium (RPMI-1640) was added to each room of the 96-well plate with an exponentially proliferating cell line. After incubation for 24 hours in a 35 ° C, 5% CO ₂ environment, the sample was diluted into 2 times in 100 mL (RPMI-1640) medium in the cell culture medium. The culture solution containing the sample was incubated at the same conditions for 48 hours. Cells were fixed with 50% trichloroacetic acid (CCl ₃ COOH) and stained with SRB dye for 30 minutes. Unbound dye was washed with 1% acetic acid. The dye bound to the protein was incubated for 5 minutes with 10 mM tris base (pH 10.5). Cell line proliferation inhibition concentration was measured at 520 nm on a microplate meter (Tecan, Salzburg, Austria).

The experimental results are shown in Table 2 below. According to the results of Table 2 below, sky tarin is three times stronger than the cancer cell line and 1.6 times more melanoma cell line than cisplatin used as an anticancer agent.

Experimental Example 2. Inhibitory Effect of Sky Tarin on Actin Polymerization

In order to observe the actin polymerization reaction of the cells, the cells were carried out in the keratinocyte line HaCaT, which is relatively large and can observe microtubules and actin (Boukemp, PB; Petrussevska, RT; Breitkreutz, D .; Hornung, J. Markham, A .; Fusenig, NE J. Cell Biol . 1988, 106 , 761). The keratinocyte line HaCaT was cultured in DMEM medium containing 10% FBS. Samples were placed in the cells and cultured overnight, and then immunofluorescence was performed to observe changes in actin. The sky tarin-treated cells were washed with ice-cold PBS solution, and then fixed with 4% formaldehyde. The cells were then incubated with primary and secondary antibodies. Microtubules were treated with antimicrotubule antibodies (Sigma, St Louis, MO) and actin was treated with fluorescent palmoydin (Sigma). Stained cells were analyzed by fluorescence microscope (Axioscope, Zeiss, Germany). Fluorescence images were taken with a Photometrics digital camera (Roper Scientific, Trenton, NJ) and attached as FIG. As a comparative compound, 2,3-diferroyl secoisolariciresinol having a chemical structure similar to that of sky tarin was used.

FIG. 5A shows the results of treatment of HaCaT cells with a sample of IC ₅₀ concentration (cytokalacin D: 1 μg / mL, sky tarin: 3.5 μg / mL, comparative compound: 12 μg / mL). 5B is a result of observing cells when HaCaT cells were treated with various concentrations of sky tarin samples.

According to Figure 5a, when the cells were treated with sky tarin, actin cytoskeleton changed into a lump-like structure in proportion to the concentration, whereas cells with the comparative compound (2,3-diferroyl secoisorarishininol) Did not significantly affect actin). According to FIG. 5B, if the treatment concentration of sky tarin is very high (12 μg / mL), all of the actin was destroyed, which is similar to the phenomenon of cytocalin D, which is well known to inhibit the polymerization of actin. . Treatment of sky tarin at low concentrations without cytotoxicity (1-3 μg / mL) also inhibited the polymerization of actin and caused separation at the cell-cell contact (Fig. 5b, 3.0 μg / mL). Arrow part). This is what happens when a cell dies. In conclusion, sky tarin inhibits the polymerization of actin, indicating a mechanism by which cells die. This is a differentiating effect that is not observed with comparative compounds (2,3-diferroyl secoisorariciresinol) with a chemical structure similar to sky tarin.

As described in detail above, the secoisorariciresinol derivative represented by Chemical Formula 1, which is characterized by the present invention, has excellent cancer cytotoxicity as a novel compound and is involved in cell growth, division, and cell-cell aggregation. By inhibiting the polymerization of actin in the body it can be usefully used for the treatment of cancer-related diseases.

1 is a mass spectrometry spectrum of empty tarin.

2 is an infrared spectroscopy spectrum of sky tarin.

3 is a ¹ H nuclear magnetic resonance spectrum of sky tarin.

4 is a ¹³ C nuclear magnetic resonance spectrum of the sky tarin.

5a and 5b are photographs confirming the actin polymerization inhibitory effect of sky tarin analyzed by immunofluorescence method,

FIG. 5A shows the results of treating HaCaT cells with a IC ₅₀ concentration sample (cytocalin D: 1 μg / mL; comparative material: 12 μg / mL; sky tarin: 3.5 μg / mL),

5B is a result of observing cells when HaCaT cells were treated with various concentrations of sky tarin samples.

Claims (7)

Secoisorariciresinol derivative represented by the following formula (1) or a pharmaceutically acceptable salt thereof: [Formula 1]

In Formula 1, R ₁ , R ₂ , and R ₃ each represent a hydrogen atom or an alkyl group having 1 to 6 carbon atoms. The compound of claim 1, wherein R ₁ , R ₂ , and R ₃ are each alkyl selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, and isobutyl. The compound of claim 1, wherein R ₁ , R ₂ , and R ₃ are each a methyl group. An anticancer agent, characterized in that the secoisorariciresinol derivative as defined in any one of claims 1 to 3 contained as an active ingredient. Extracting the sky seed pulverized product into a single or mixed solvent selected from the group of organic solvents consisting of alcohols, ethyl acetate and hexanes having 1 to 6 carbon atoms; Dissolving the solvent extract in an alcohol having 1 to 6 carbon atoms, extracting with hexane and concentrating to obtain an alcohol layer; Primary silica gel column chromatography on the concentrate to obtain an active fraction (R _f = 0.4 to 0.6, eluent: methylene chloride / methanol / water, 90: 9.5: 0.5 v / v / v); A process of obtaining the active fraction (R _f = 0.5-0.6, eluent: methylene chloride / methanol / water, 90: 9.5: 0.5 v / v / v) by performing secondary silica gel column chromatography on the obtained active fraction; And The active fraction obtained by the secondary silica gel column chromatography was subjected to C18 reversed phase high pressure liquid chromatography, and the 1- O -ferroyl secoisolaryrrishinol represented by the following Chemical Formula 1a (R _t = 37.2 min, 30-60% water soluble) Acetonitrile, 60 min, 7 mL / min, UV 225 nm) Separation method of secoisolarishilsinol derivatives as a natural product, characterized in that it comprises. [Formula 1a]

The method of claim 5, wherein the alcohol is selected from methanol, ethanol, n-propanol, isopropanol, n-butanol, and isobutanol. The method of claim 5, wherein the alcohol is an anhydrous alcohol or a water-soluble alcohol containing water in the range of 1 to 80% by volume.

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