KR20200008424A - Composition for preventing or treating cancer comprising glycosyl glycerides from Malva verticillata - Google Patents

Abstract

The present invention relates to: a pharmaceutical composition and a food composition for preventing or treating cancer, wherein compositions contain a Malva verticillata extract, a fraction thereof, a compound represented by chemical formula 1 isolated from the same, or pharmaceutically acceptable salt of the compound; and a method for manufacturing the compound. The Malva verticillata extract, the fraction thereof, and the compound represented by chemical formula 1 isolated from the same exhibit cytotoxicity on cancer cells and induce cell death, thereby being able to be utilized as the pharmaceutical composition, the food composition and the like for preventing, alleviating, or treating cancer.

Description

Composition for preventing or treating cancer comprising glycosyl glycerides derived from mallow {Composition for preventing or treating cancer comprising glycosyl glycerides from Malva verticillata}

The present invention Malva verticillata ( Malva verticillata ) extract, fractions thereof, a compound of formula 1 isolated from them or a pharmaceutical composition for cancer prevention or treatment comprising a pharmaceutically acceptable salt of the compound as an active ingredient; Food compositions; And methods for preparing the compounds.

Cancer is one of the high mortality diseases worldwide. Currently cancer therapies such as chemotherapy, surgery and radiation therapy have been developed, but there are a number of side effects and costs associated with their application. Epidemiological evidence has shown that eating certain foods can potentially prevent the development of cancer by up to 35%. In this regard, bioactive ingredients and diets present in nature provide an essential link to the maintenance and promotion of health as well as the prevention of chronic diseases including cancer. The most practical way to reduce the incidence and mortality of cancer is to delay the oncogenesis process using chemical cancer prevention agents. This requires safer compounds, especially those derived from natural raw materials for chemical cancer prevention.

Malva verticillata (Chinese mallow ) is one of the oldest plants cited in the written literature, a leaf vegetable that has long been used in Korea and China. The seeds of mallow have been used as a medicine for diuretics, laxatives and lactose in herbal medicine. Polysaccharides, the major component of mallow seeds, have been reported to exhibit anti-complementary, immunological and hypoglycemic activity. On the other hand, mallow leaf has also been used as a medicament for the treatment of gastrointestinal and respiratory diseases in Chinese medicine. However, there has been little research on the chemical composition of mallow leaves and their biological activities.

Under these circumstances, the present inventors have diligently identified and identified glycosyl glyceride compounds from mallow above ground fractions, as a result of diligent efforts to find chemical cancer prevention agents derived from natural raw materials, among which (2S) -1-O- (6 -Deoxy-6-sulfo) -α-D-glucopyranosyl-2-O-linolenoyl-3-O-palmitoyl glyceride; (2S) -1-O- (6-deoxy-6-sulfo) -α-D-glucopyranosyl-2,3-di-O-linolenoyl glyceride; Or (2S) -1-O-6'-0- (aD-galactopyranosyl) -β-D-galactopyranosyl-2,3-di-O-palmitoyl glyceride is applied to cancer cells The present invention was completed by confirming toxicity and inducing apoptosis.

One object of the present invention is a pharmaceutical composition for cancer prevention or treatment comprising mallow extract, fractions thereof, the compound represented by the formula (1) isolated from them or a pharmaceutically acceptable salt of the compound as an active ingredient, R ₁ is -H, linolenoyl, palmitoyl or stearoyl; R ₂ is linolenoyl, palmitoyl or stearoyl; R ₃ is to provide a pharmaceutical composition, which is —SO ₃ ^— , —OH or glycosyl.

[Formula 1]

Another object of the present invention is to provide a food composition comprising the active ingredient.

Another object of the present invention is a novel compound derived from mallow extract or fraction, (2S) -1-O-β-D-galactopyranosyl-3-O-isostearoyl glyceride ((2S) -1-O-β-D-galactopyranosyl-3-O-isostearoyl glyceride, compound 5).

Another object of the present invention to provide a method for preparing a compound represented by the formula (1) comprising the step of separating the compound represented by the formula (1) from the mallow extract or fraction.

This will be described in detail below. In addition, each description and embodiment disclosed in this application may apply to each other description and embodiment. That is, all combinations of the various elements disclosed in this application are within the scope of the present application. In addition, the scope of the present application is not to be limited by the specific description described below.

One aspect of the present invention for achieving the above object is a mallow extract, fractions thereof, cancer prevention or treatment comprising a compound represented by the following formula (1) isolated from them or a pharmaceutically acceptable salt of the compound as an active ingredient As a pharmaceutical composition for

[Formula 1]

In the general formula 1 R ₁ is -H, linoleate leno days, palmitoyl or stearoyl one (stearoyl), and; R ₂ is linolenoyl, palmitoyl or stearoyl; R ₃ provides a pharmaceutical composition, which is —SO ₃ ^— , —OH or glycosyl.

Specifically, the fraction may be n-butanol fraction of mallow, the compound is (2S) -1-O- (6-deoxy-6-sulfo) -α-D-glucopyranosyl-2-O- Linolenoyl-3-O-palmitoyl glyceride (Compound 1), (2S) -1-O- (6-deoxy-6-sulfo) -α-D-glucopyranosyl-2,3-di- O-linolenoyl glyceride (Compound 2), (2S) -1-O-β-D-galactopyranosyl-3-O-palmitoyl glyceride (Compound 3), (2S) -1-O- β-D-galactopyranosyl-3-O-stearoyl glyceride (Compound 4), (2S) -1-O-β-D-galactopyranosyl-3-O-isostearoyl glyceride (Compound 5), (2S) -1-O-β-D-galactopyranosyl-3-O-linolenoyl glyceride (Compound 6), (2S) -1-O-β-D-galacto Pyranosyl-2,3-di-O-palmitoyl glyceride (Compound 7), (2S) -1-O-β-D-galactopyranosyl-2,3-di-O-linolenoyl glyceride (Compound 8), (2S) -1-O-6'-O- (α-D-galactopyranosyl) -β-D-galactopyranosyl-3-O-palmitoyl Ceride (Compound 9), (2S) -1-O-6'-O- (α-D-galactopyranosyl) -β-D-galactopyranosyl-3-O-linolenoyl glyceride (compound 10), (2S) -1-O-6'-O- (α-D-galactopyranosyl) -β-D-galactopyranosyl-2,3-di-O-palmitoyl glyceride (compound 11), (2S) -1-O- (6-O-α-D-galactopyranosyl) -β-D-galactopyranosyl-2-O-stearoyl-3-O-linolenoyl Glyceride (Compound 12) and (2S) -1-O- (6-O-α-D-galactopyranosyl) -β-D-galactopyranosyl-2,3-di-O-linolenoyl -One or more compounds selected from the group consisting of glycerides (compound 13), more specifically (2S) -1-O- (6-deoxy-6-sulfo) -α-D-glucopyranosyl- 2-O-linolenoyl-3-O-palmitoyl glyceride (Compound 1); (2S) -1-O- (6-deoxy-6-sulfo) -α-D-glucopyranosyl-2,3-di-O-linolenoyl glyceride (Compound 2); Or (2S) -1-O-6'-0- (α-D-galactopyranosyl) -β-D-galactopyranosyl-2,3-di-O-palmitoyl glyceride (Compound 11) It may be, but is not limited thereto.

Compounds 1 and 2 include 6-deoxy-6-sulfo saccharide ((6-deoxy-6-sulfo) -α-D-glucopyranose) and are rarely found from natural raw materials. The anticancer effect of is not known in the past, Compound 11 showed the best anticancer effect among the compounds of the present invention, the anticancer effect of mallow-derived compounds of the present invention, including compounds 1, 2 and 11 was first confirmed in the present invention .

That is, the present invention shows that not only mallow extracts and fractions but also compounds including compounds 1, 2 and 11 isolated and identified therefrom exhibit excellent anticancer activity against various cancer cells such as HepG2, AGS, HCT-15, and A549. Based on the technical idea, this was first identified by the inventors.

In one embodiment of the present invention, a fraction of mallow or a compound isolated therefrom, specifically an n-butanol fraction of mallow, a compound of formula 1 shows cytotoxicity against cancer cells and expression of proteins associated with apoptosis It has been shown that the level is adjusted to induce apoptosis of cancer cells, the fraction or compound has a prophylactic or therapeutic effect of cancer.

In the present invention, " Malva verticillata ( Chinese mallow)" is a dicotyledonous plant belonging to the mallow family (Mavales) Malvaceae. It is known to use, but its anticancer effect is not known at all. The mallow may be purchased commercially, or may be used collected or grown in nature. Specifically, the roots, stems, leaves, etc. of the mallow may be used without limitation, and more specifically, the entire upper part of the mallow may be used, but is not limited thereto.

In the present invention, the "extract" is the extract itself and the extract, such as the extract obtained by extracting the mallow, the dilution or concentrate of the extract, the dried product obtained by drying the extract, the crude or purified product of the extract, or a mixture thereof It includes extracts of all formulations that can be formed using.

The kind of extraction solvent used to extract the extract in the present invention is not particularly limited, and any solvent known in the art may be used. Non-limiting examples of such extraction solvents include water; C1-C4 lower alcohols, such as methanol, ethanol, propyl alcohol, and butyl alcohol; Polyhydric alcohols such as glycerin, butylene glycol and propylene glycol; And hydrocarbon solvents such as methyl acetate, ethyl acetate, acetone, benzene, hexane, diethyl ether and dichloromethane; Or a mixture thereof may be used, specifically methanol may be used, but is not limited thereto.

The method of extracting the extract in the present invention is not particularly limited and may be extracted according to a method commonly used in the art. Non-limiting examples of the extraction method, hot water extraction method, ultrasonic extraction method, filtration method, reflux extraction method and the like, these may be performed alone or in combination of two or more methods.

"Fragment" in the present invention means the result obtained by the fractionation method for separating a specific component or a specific group from a mixture comprising various components. Specifically, the present invention includes the result of fractionating the mallow extract by various methods such as solvent fractionation, ultrafiltration fractionation, chromatography fractionation, and the like, more specifically, n-butanol fraction of the mallow extract, This is not restrictive.

In one embodiment of the present invention, the ground portion of the dried mallow was extracted and filtered with 80% methanol at room temperature, reduced pressure and evaporated to obtain an extract, from which water, ethyl acetate (EtOAc) and n-butanol (n- BuOH) fraction was obtained. The m-butanol fraction of the above-ground surface shows a significantly superior cytotoxicity against cancer cells compared to the fractions of water and ethyl acetate, it was confirmed that there is an effect of inducing apoptosis. In addition, glycosyl glyceride compounds 1 to 13 were isolated and identified from the n-butanol fraction, and the anticancer effects of each compound were confirmed. In particular, it was confirmed that Compounds 1, 2, and 11 exhibited remarkably good cytotoxicity against cancer cells and had an effect of inducing apoptosis.

In addition, the compounds derived from mallow extracts or fractions of the present invention may be isolated from other natural sources, and may be chemically synthesized or commercially available through methods known in the art, and related to the available route It can be used in the present invention without.

In the present invention, "pharmaceutically acceptable salt" means a salt in a form that can be used pharmaceutically even among salts in which cations and anions are bound by electrostatic attraction, and are relatively nontoxic and harmless to the patient. Concentrations at which the adverse effects attributable to this salt do not reduce any beneficial organic or inorganic addition salts of the compounds from the mallow extracts or fractions of the present invention, specifically compounds 1, 2 and 11 it means. Such salts include acid addition salts formed by pharmaceutically acceptable free acids or metal salts formed by bases.

Organic acids and inorganic acids may be used as the free acid, hydrochloric acid, phosphoric acid, sulfuric acid, nitric acid, tartaric acid, etc. may be used as the inorganic acid, and methanesulfonic acid, p-toluenesulfonic acid, acetic acid, trifluoroacetic acid, and maleic acid may be used as the organic acid. (maleic acid), succinic acid, oxalic acid, benzoic acid, tartaric acid, fumaric acid, manderic acid, propionic acid, citric acid, lactic acid, glycolic acid, gluconic acid (gluconic acid), galacturonic acid, glutamic acid, glutaric acid, glucuronic acid, glucuronic acid, aspartic acid, ascorbic acid, carbonic acid, vanic acid, hydroiodic acid, etc. This is not restrictive.

Bases can also be used to make pharmaceutically or food acceptable metal salts. Alkali metal salts or alkaline earth metal salts are obtained, for example, by dissolving a compound in an excess of alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the insoluble compound salt, and then evaporating and drying the filtrate. In this case, as the metal salt, it is particularly suitable to prepare sodium, potassium, or calcium salt in a pharmaceutical or food form, but is not limited thereto. Corresponding silver salts can also be obtained by reacting an alkali or alkaline earth metal salt with a suitable silver salt (eg, silver nitrate).

Such pharmaceutically acceptable salts include salts of acidic or basic groups which may be present in the compounds derived from mallow extracts or fractions of the invention, unless otherwise specified. For example, pharmaceutically or food acceptable salts may include sodium, calcium and potassium salts of the hydroxy group, and other pharmaceutically acceptable salts of the amino group include hydrobromide, sulfate, hydrogen sulfate, phosphate , Hydrogen phosphate, dihydrogen phosphate, acetate, succinate, citrate, tartrate, lactate, mandelate, methanesulfonate (mesylate) and p-toluenesulfonate (tosylate) salts, and the like. It may be prepared through a known method for preparing a salt.

In the present invention, "cancer" refers to a tumor in which normal tissue cells are formed by the action or factors of various physical, chemical, and biological cancerous substances, and the cancer includes, but is not limited to, the type of cancer. Esophageal cancer, stomach cancer, colon cancer, rectal cancer, oral cancer, pharyngeal cancer, laryngeal cancer, lung cancer, colon cancer, breast cancer, cervical cancer, endometrial cancer, ovarian cancer, prostate cancer, testicular cancer, bladder cancer, kidney cancer, liver cancer, pancreatic cancer, bone cancer, connective tissue Cancer, skin cancer, brain cancer, thyroid cancer, leukemia, Hodgkin's disease, lymphoma or multiple myeloma hematologic cancer, specifically gastric cancer, lung cancer, colorectal cancer, or liver cancer, more specifically gastric cancer, but is not limited thereto. It doesn't happen.

In one embodiment of the present invention, the compound of the present invention, including the m-butanol fraction of the mallow, Compounds 1, 2, and 11 shows excellent anticancer effects against various cancer cells such as HepG2, AGS, HCT-15, and A549. Confirmed.

"Prevention" in the present invention means any action that delays the onset or progression of cancer by administration of the composition of the present invention.

In the present invention, "treatment" means any action that improves or advantageously changes the symptoms of the onset of cancer by administration of the composition of the present invention.

"Improvement" in the present invention means any action that at least reduces the degree of symptoms of cancer, increases cell death of cancer cells or inhibits proliferation.

As used herein, the term "pharmaceutical composition" means prepared for the purpose of preventing or treating a disease, and may be administered in various oral and parenteral dosage forms in actual clinical administration, and when formulated, commonly used fillers, Diluents or excipients such as extenders, binders, wetting agents, disintegrants, surfactants and the like can be prepared.

In addition, each formulation may further include a pharmaceutically acceptable additive, wherein the pharmaceutically acceptable additives include starch, gelatinized starch, microcrystalline cellulose, lactose, povidone, colloidal silicon dioxide, calcium hydrogen phosphate , Lactose, mannitol, syrup, gum arabic, pregelatinized starch, corn starch, powdered cellulose, hydroxypropyl cellulose, oppadry, sodium starch glycolate, carnauba lead, synthetic aluminum silicate, stearic acid, magnesium stearate, aluminum stearate, Calcium stearate, sucrose, dextrose, sorbitol, talc and the like can be used.

Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and such solid preparations may contain at least one excipient such as starch, calcium carbonate, water, or the like in the mixed extract of the present invention. Sucrose, lactose or gelatin can be mixed and prepared. In addition to the simple excipients, lubricants such as magnesium styrate talc may also be used. Liquid preparations for oral administration include suspensions, solvents, emulsions and syrups, and include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. Can be.

Formulations for parenteral administration may include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and the suspension solvent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

The composition of the present invention can be administered orally or parenterally according to the desired method, and during parenteral administration, external skin or intraperitoneal injection, rectal injection, subcutaneous injection, intravenous injection, intramuscular injection or intrathoracic injection Can be selected. The dosage may vary depending on the weight, age, sex, health condition, diet, time of administration, method of administration, rate of excretion and severity of the patient.

The composition of the present invention may be administered in a pharmaceutically effective amount. The pharmaceutically effective amount refers to an amount sufficient to treat the disease at a reasonable benefit / risk ratio applicable to the medical treatment and not causing side effects, and the effective dose level refers to the patient's state of health, type of disease, severity, The activity of the drug, the sensitivity to the drug, the method of administration, the time of administration, the route of administration and the rate of excretion, the duration of treatment, the factors including the drug used in combination or co-administration, and other factors well known in the medical art. In addition, the pharmaceutical compositions of the present invention may be used alone or in combination with other pharmaceutically active compounds which have a prophylactic, therapeutic, or ameliorating effect of cancer or in suitable collections.

The pharmaceutical compositions of the present invention may be administered as individual therapeutic agents or in combination with other therapeutic agents and may be administered sequentially or simultaneously with conventional therapeutic agents. And single or multiple administrations. Taking all of these factors into consideration it is important to administer an amount that can achieve the maximum effect in a minimum amount without causing side effects and can be readily determined by one skilled in the art.

Another aspect of the present invention for achieving the above object comprises a mallow extract, a fraction thereof, a compound represented by the formula (1) isolated from them or a pharmaceutically acceptable salt of the compound as an active ingredient, cancer prevention Or a therapeutic food composition. The mallow, extract, fraction and cancer are as described above.

In the present invention, the term "food" includes, for example, various foods, beverages, gums, teas, vitamin complexes, health functional foods, and the like, and includes all foods in a general sense. The food may be prepared in the form of tablets, granules, powders, capsules, liquid solutions and pills according to known production methods, and the content of the extract according to the present invention is 0.0001 based on the total weight of the food composition according to the formulation. It can be adjusted to 100% by weight. There is no particular limitation on other components except including mallow extracts, fractions or compounds derived therefrom as an active ingredient, and various conventional flavoring agents or natural carbohydrates may be included as additional ingredients.

The term "health functional (sex) food" is the same term as food for special health use (FOSHU), means a food with a high medical effect, processed to ensure that the bioregulatory function appears efficiently in addition to nutrition supply do. The term "functionality" means obtaining a useful effect for health purposes such as nutrient control or physiological action on the structure and function of the human body. The health functional food of the present invention can be prepared by a method commonly used in the art, and the preparation can be prepared by adding raw materials and ingredients commonly added in the art. In addition, the formulation of the health functional food can also be prepared without limitation as long as the formulation is recognized as a food. The health functional food composition of the present invention can be prepared in various forms of formulations, unlike general medicines, there is no side effect that may occur when taking a long-term use of the drug as a raw material, there is an excellent portability.

Specifically, the health functional food is a food prepared by adding the composition of the present invention to food materials, such as beverages, teas, spices, gums, confections, or the like, encapsulated, powdered, suspensions, etc. Means to bring the effect, unlike the general medicine has the advantage that there is no side effect that can occur when taking long-term use of the drug as a raw material.

The dietary supplement composition may further include a physiologically acceptable carrier, and the type of carrier is not particularly limited and may be any carrier that is commonly used in the art. In addition, the health functional food composition may include any additional ingredients that are commonly used in the food composition to improve the smell, taste, time and the like, may be selected according to the type of food and used in an appropriate amount.

Another aspect of the present invention for achieving the above object is a novel compound, (2S) -1-O-β-D-galactopyranosyl-3-O-isostearoyl glyceride ((2S)- 1-O-β-D-galactopyranosyl-3-O-isostearoyl glyceride, compound 5). The compound may be derived from mallow extracts or fractions thereof. The mallow, extract, fractions are as described above.

Another aspect of the present invention for achieving the above object provides a method for preparing a compound represented by the formula (1) comprising the step of separating the compound represented by the formula (1) from the mallow extract or fraction. The mallow, extract, fraction and the compound represented by the formula (1) are as described above.

The mallow extract of the present invention, the fractions thereof and the compound represented by the formula (1) isolated therefrom exhibits cytotoxicity to various cancer cells and induces apoptosis, pharmaceutical compositions, food compositions for the prevention, improvement or treatment of cancer It can be used as such.

Figure 1 shows the structure of glycosyl glyceride compounds 1 to 13 isolated from the mallow ground in the present invention.
FIG. 2 shows the results of apoptotic analysis when n-butanol fractions of mallow, compounds 1, 2 and 11 were treated with AGS (human gastric cancer cells).
FIG. 3 shows the protein (PARP, caspase-3, Bax and Bcl-2) associated with apoptosis when treated with AGS (human gastric cancer cells) with n-butanol fraction of mallow, Compounds 1, 2 and 11 The result of measuring the expression level is shown.

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, these examples are for illustrative purposes only and the scope of the present invention is not limited only to these examples.

Example 1: Materials, Reagents and Instruments

1-1: Plant Material

The ground portion of Malva verticillata used in this example was collected at a commercial farm in Namyangju, South Korea in April 2016, and a specimen sample (KHU20160419) is stored in a Natural Product Chemistry laboratory at Kyung Hee University.

1-2: Reagents and Instruments

Silica gel (SiO ₂ ) and octadecyl SiO ₂ (ODS) resins used in column chromatography (CC) were Kiesel gel 60 (Merck, Darmstadt, Germany) and Lichroprep RP-18 (40-60 μm, Merck), respectively. Thin layer chromatography (TLC) analysis was performed using Kiesel gel 60 F ₂₅₄ and RP-18 F _254S (Merck) TLC plates and the compounds were Spectroline Model ENF-240 C / F (Spectronics Corporation, Westbury, NY, USA) Detection was done using a UV lamp and 10% H ₂ SO ₄ solution. Nuclear magnetic resonance (NMR) spectra were recorded using a 400 MHz FT-NMR spectrometer (Varian Inova AS-400, Palo Alto, Calif., USA). Deuterium solvents are available from Merck Co. Ltd and Sigma Aldrich Co. Ltd. (St, Louis, MO, USA). IR spectra were obtained from the Perkin Elmer Spectrum One FT-IR spectrometer (Buckinghamshire, England). Fast atom bombardment mass spectrometry (FAB-MS) was recorded by JEOL JMS-700 (Tokyo, Japan). Gas chromatography mass spectrometry (GC-MS) was recorded by Shimadzu GC-MSQP2010 Plus (Kyoto, Japan). Optical rotation was measured with a JASCO P-1010 digital polarimeter (Jasco, Tokyo, Japan).

Example 2: Preparation of Mallow Extracts and Fractions

The dry mallow ground portion (3.1 kg) was extracted with 80% methanol (MeOH, 54.0L Х 5) at room temperature for 24 hours. The extract was filtered through filter paper and evaporated at 43 ° C. under reduced pressure to yield 794 g of extract.

The resulting methanol extract was suspended in water (2 L) and extracted successively with ethyl acetate (EtOAc, 2 L Х 4) and n-butanol (n-BuOH, 2 L Х 4). Each layer was concentrated to give an ethyl acetate fraction (MVE, 80 g), n-butanol fraction (MVB, 77 g) and water fraction (MVW, 637 g).

Example 3: Isolation and Identification of Compounds from the n-Butanol Fraction of Mallow

3-1: Separation of Compounds from the n-Butanol Fraction of Mal

The n-butanol fraction (MVB, 77 g) of Example 2 was subjected to silica gel column chromatography (CC) (φ 10 Х 15 cm), and chloroform (CHCl ₃ ) -methanol (MeOH) -water (H ₂ O) (25 : 3: 1 → 22: 3: 1 → 20: 3: 1 → 18: 3: 1 → 15: 3: 1 → 12: 3: 1, each of which was eluted with 14 fractions (MVB-1). To MVB-14).

Fraction MVB-6 (3.6 g, elution volume / total volume (V _e / V _t ) 0.053-0.096) was applied to ODS CC (φ4.5 Х 6 cm) and acetone-water (3: 2 → 2: Eluting with 3.6 L) gave 11 fractions (MVB-6-1 to MVB-6-11). Among them, fraction MVB 6-4 (659.4 mg, V _e / V _t 0.031-0.092) was applied to silica gel CC (φ2.5 Х 15 cm) and washed with chloroform-methanol-water (20: 3: 1, 1.4 L). Eluted, purified with 11 fractions (MVB-6-4-1 to MVB-6-4-11) Compound 3 [MVB6-4-10, 21.2 mg, V _e / V _t 0.286-0.514, TLC ( Kiesel gel 60 F254) R _f 0.38, chloroform-methanol-water (10: 3: 1), TLC (RP-18 F254S) Rf 0.74, methanol-water (3: 1)].

Fraction MVB-7 (5.0 g, V _e / V _t 0.098-0.126) was applied to ODS CC (φ4.5 Х 5 cm) and methanol-water (1: 1 → 2: 1 → 6: 1 → 8: 1, Isolated by elution with 3.8 L each) and purified with 22 fractions (MVB-7-1 to MVB-6-22) 5 [MVB-7-9, 84.2 mg, V _e / V _t 0.212- 0.217, TLC (Kiesel gel 60 F254) R _f 0.90, Chloroform-methanol-water (7: 3: 1), TLC (RP-18 F254S) Rf 0.70, Methanol-water (5: 1)], Compound 6 [MVB -7-12, 142.8 mg, V _e / V _t 0.323-0.366, TLC (Kiesel gel 60 F254) R _f 0.90, Chloroform-methanol-water (7: 3: 1), TLC (RP-18 F254S) Rf 0.41 , Methanol-water (5: 1)], compound 12 [MVB-7-18, 34.0 mg, V _e / V _t 0.737-0.750, TLC (Kiesel gel 60 F254) R _f 0.90, chloroform-methanol-water (7 : 3: 1), TLC (RP-18 F254S) Rf 0.52, Methanol-water (14: 1)], and Compound 13 [MVB-7-20, 586.2 mg V _e / V _t 0.787-0.842, TLC (Kiesel gel 60 F254) R _f 0.90, chloroform-methanol-water (7: 3: 1), TLC (RP-18 F254S) Rf 0.24, methanol-water (14: 1)].

Fraction MVB-8 (3.5 g, V _e / V _t 0.128-0.383) was applied to ODS CC (φ4.5 Х 8 cm) and methanol-water (3: 1 → 4: 1 1 5: 1 1 6: 1, Each fraction was eluted with 1.0 L) to obtain 17 fractions (MVB-8-1 to MVB-8-17). Of these, 13 fractions were applied by MVB-8-17 (879.3 mg, V _e / V _t 0.602-1.000) to ODS CC (φ4 Х 8 cm) and eluted with acetone-water (3: 1, 3.0 L). Compound 8 purified with (MVB-8-17-1 to MVB-8-17-13) [MVB-8-17-10, 26.0 mg, V _e / V _t 0.480-0.580, TLC (Kiesel gel 60 F254 ) R _f 0.35, chloroform-methanol-water (10: 3: 1), TLC (RP-18 F254S) R _f 0.20, methanol-water (2: 1)].

Fraction MVB-9 (4.0 g, V _e / V _t 0.385-0.557) was applied to ODS CC (φ4 Х 6 cm) and methanol-water (3: 2 → 2: 1 → 2.5: 1 → 3: 1 → 5: Eluting with 3.6 L) each gave 16 fractions (MVB-9-1 to MVB-9-16) with compound 2 [MVB-9-10, 44.4 mg V _e / V _t 0.602-0.668 , TLC (Kiesel gel 60 F254) R _f 0.25, chloroform-methanol-water (8: 3: 1), TLC (RP-18 F254S) R _f 0.28, methanol-water (5: 1)].

Fraction MVB-9-5 (383.9 mg, V _e / V _t 0.116-0.263) was applied to ODS CC (φ2.5 Х 6 cm) and eluted with acetone-water (2: 3, 4.0 L) to give 14 fractions. Compound 11 purified with (MVB-9-5-1 to MVB-9-5-14) [MVB-9-5-9, 96.3 mg V _e / V _t 0.602-0.668, TLC (Kiesel gel 60 F254) R _f 0.25, chloroform-methanol-water (8: 3: 1), TLC (RP-18 F254S) R _f 0.48, acetone-water (2: 1)].

Fraction MVB-11 (8.7 g, V _e / V _t 0.560-0.755) was applied to ODS CC (φ4.5 Х 8 cm) and methanol-water (2: 1 → 4: 1 → 6: 1, 3.0L each) Compound eluted with 16 fractions (MVB-11-1 to MVB-11-16) as a result of elution with 10 [MVB-11-10, 732.4 mg V _e / V _t 0.582-0.629, TLC (Kiesel gel 60 F254) R _f 0.71, chloroform-methanol-water (65:35:10), TLC (RP-18 F254S) R _f 0.18, methanol-water (4: 1)].

Fraction MVB-11-6 (1.5 g, V _e / V _t 0.102-0.289) was applied to silica gel CC (φ3.5 Х 15 cm) and eluted with chloroform-methanol (5: 2 → 2: 1, all 1.5L). Separation resulted in 10 fractions (MVB-11-6-1 to MVB-11-6-10). Among them, fraction MVB-11-6-5 (263.0 mg, V _e / V _t 0.240-0.473) was applied to ODS CC (φ3 Х 5 cm) and methanol-water (3: 2 → 1: 1, both 1.8L Was purified by elution) and purified with 12 fractions (MVB-11-6-5-1 to MVB-11-6-5-12) 9 [MVB-11-6-5-10, 38.8 mg V _e / V _t 0.502-0.746, TLC (Kiesel gel 60 F254) R _f 0.18, chloroform-methanol (2: 1), TLC (RP-18 F254S) R _f 0.50, methanol-water (3: 1)] Obtained.

Fraction MVB-11-16 (1.8 g, V _e / V _t 0.998-1.000) was applied to silica gel CC (φ4 Х 15 cm) and ethyl acetate (EtOAc) -n-butanol (n-BuOH) -water (7: 3 Eluting with 1: 2: 5: 1, all 3L) gave 14 fractions (MVB-11-16-1 to MVB-11-16-14). Among them, fraction MVB-11-16-7 (99.0 mg, V _e / V _t 0.120-0.237) was applied to ODS CC (φ2.5 Х 5 cm) and eluted with acetone-water (2: 1, 0.46 L). Compound 1 [MVB-11-16-7-5, 23.8 mg V _e purified with 12 fractions (MVB-11-16-7-1 to MVB-11-16-7-12) / V _t 0.110-0.209, TLC (Kiesel gel 60 F254) R _f 0.85, Chloroform-methanol-water (6: 4: 1), TLC (RP-18 F254S) R _f 0.46, Acetone-water (3: 1) ] Was obtained.

Fraction MVB-12 (11.0 g, V _e / V _t 0.756-0.898) was applied to ODS CC (φ6.5 Х 5 cm) and methanol-water (1: 1 → 2: 1 → 3: 1 → 5: 1, Isolated by elution with 3.8 L each), purified with 18 fractions (MVB-12-1 to MVB-12-18) 4 [MVB-12-10, 632.4 mg V _e / V _t 0.303-0.349 , TLC (Kiesel gel 60 F254) R _f 0.50, chloroform-methanol-water (65:35:10), TLC (RP-18 F254S) R _f 0.78, methanol-water (5: 1)].

Water fraction (MVW, 635 g) was applied to an anionic HP-20 column (φ9 Х 30 cm) and methanol-water (0: 1 1 → 1: 4 → 2: 3 → 3: 2 → 4: 1 1: 1: 0, respectively) Elution with 6.0 L) gave 9 fractions (MVW-1 to MVW-9). Among them, fraction MVW-7 (1.0 g, V _e / V _t 0.911-0.967) was applied to silica gel CC (φ4.5 Х 15 cm) and chloroform-methanol (40: 1 1 → 15: 1 1 10: 1 1 → 5 : 1, eluted with 5.6 L each, and the resulting compound was purified with 23 fractions (MVW-7-1 to MVW-7-23). 7 [MVW-7-18, 22.8 mg V _e / V _t 0.476-0.500, TLC (Kiesel gel 60 F254) R _f 0.50, chloroform-methanol (5: 1), TLC (RP-18 F254S) R _f 0.82, acetone-water (4: 1)].

3-2: Identification of Isolated Compounds 1 to 13

The compounds 1 to 13 obtained in Example 3-1 were subjected to NMR, IR, FAB-MS and GC-MS analysis to identify the structure of each compound. Specifically, ¹ H (400 MHz) and ¹³ C (100 MHz) NMR results of Compounds 1, 2, and 5 are shown in Table 1 below.

TABLE 1

The result of analyzing compound 1 is as follows.

Pale yellow wax

[a] D + 30.8 ° (c 0.10, MeOH)

IR (KBr, m) 3390, 1735, 1612, 1186 cm ^-1

-negative FAB-MS m / z 815 [MH] ^- , m / z 559 [MHR ₃ ] ^- , m / z 537 [MHR ₂ ]-, m / z 277 [R2-H] ^- , m / z 255 [ R ₃ -H] ^- , m / z 225 [the dehydrated product of the (6-deoxy-6-sulfo) -D-glucopyranose ion m / z 243], m / z 80 [SO ₃ ] ^-

negative HR-FAB-MS m / z 815.4982 [MH] ^- (calcd for C ₄₃ H ₇₅ O ₁₂ S-, 815.4985)

GC-MS t _R = 10.27 min (palmitic acid methyl ester), 13.76 min (linolenic acid methyl ester).

As a result, Compound 1 was prepared as (2S) -1-O- (6-deoxy-6-sulfo) -α-D-glucopyranosyl-2-O-linolenoyl-3-O-palmitoyl glyceride ( (2S) -1-O- (6-deoxy-6-sulfo) -α-D-glucopyranosyl-2-O-linolenoyl-3-O-palmitoyl glyceride). Compound 1 is the first compound identified and identified from natural sources.

The result of analyzing Compound 2 is as follows.

Pale yellow wax

-[a] D + 28.4 ° (c 0.10, MeOH)

IR (KBr, v) 3386, 1732, 1610, 1178 cm ^-1

positive FAB-MS m / z 861 [M + Na] ⁺

negative FAB-MS m / z 837 [M-H]-, m / z 559

-[MHR _{2 or 3} ]-, m / z 277 [R ₂ or ₃ -H]-, m / z 225 [the dehydrated product of the (6-deoxy-6-sulfo) -D-glucopyranose ion m / z 243], m / z 80 [SO ₃ ] ^- , negative HR-FAB-MS m / z 837.4822 [M ⁻ H] ⁻ (calcd for C ₄₅ H ₇₃ O ₁₂ S ⁻ , 837.4828)

GC-MS t _R = 13.76 min (linolenic acid methyl ester).

As a result, Compound 2 was prepared as (2S) -1-O- (6-deoxy-6-sulfo) -α-D-glucopyranosyl-2,3-di-O-linolenoyl glyceride ((2S) -1-O- (6-deoxy-6-sulfo) -α-D-glucopyranosyl-2,3-di-O-linolenoyl Glyceride). Compound 2 is the first compound identified and identified from natural sources.

The result of analyzing compound 3 is as follows.

-Pale yellow wax

[a] D + 9.3 ° (c 0.10, MeOH)

IR (KBr, m) 3379, 1725 cm ^-1

positive FAB-MS m / z 515 [M + Na] ⁺

GC-MS t _R = 11.06 min (palmitic acid).

As a result, Compound 3 was prepared as (2S) -1-O-β-D-galactopyranosyl-3-O-palmitoyl glyceride ((2S) -1-O-β-D-galactopyranosyl-3-O- palmitoyl glyceride).

The result of analyzing Compound 4 is as follows.

-Pale yellow wax

[a] D + 2.6 ° (c 0.10, MeOH)

IR (KBr, m) 3383, 1723 cm ^-1

positive FAB-MS m / z 543 [M + Na] ⁺

GC-MS t _R = 14.36 min (stearic acid methyl ester)

As a result, Compound 4 was prepared as (2S) -1-O-β-D-galactopyranosyl-3-O-stearoyl glyceride ((2S) -1-O-β-D-galactopyranosyl-3-O -stearoyl glyceride) was confirmed.

The result of analyzing compound 5 is as follows.

-Pale yellow wax

[a] D + 5.5 ° (c 0.10, MeOH)

IR (KBr, m) 3380, 1722 cm ^-1 ;

negative FAB-MS m / z 555 [M + Cl] ^- , m / z 339 [MH-180 (hexose + H ₂ O)] ^- , m / z 283 [R ₃ -H] ^- ,

negative HR-FAB-MS m / z 555.3289 [M + Cl] ^- (calcd for C ₂₇ H ₅₂ O ₉ Cl, 555.3299)

GC-MS t _R = 14.36 min (isostearic acid methyl ester)

As a result, Compound 5 was prepared as (2S) -1-O-β-D-galactopyranosyl-3-O-isostearoyl glyceride ((2S) -1-O-β-D-galactopyranosyl-3- O-isostearoyl glyceride) was confirmed. Compound 5 is a novel glycosyl glyceride compound, named Malvaglycolipid A.

The result of analyzing compound 6 is as follows.

-Pale yellow wax

[a] D + 3.7 ° (c 0.10, MeOH)

IR (KBr, m) 3386, 1722, 1612 cm ^-1

positive FAB-MS m / z 537 [M + Na] ⁺

GC-MS t _R = 13.76 min (linolenic acid methyl ester)

As a result, Compound 6 was dissolved in (2S) -1-O-β-D-galactopyranosyl-3-O-linolenoyl glyceride ((2S) -1-O-β-D-galactopyranosyl-3-O -linolenoyl glyceride) was confirmed.

The result of analyzing Compound 7 is as follows.

-Pale yellow wax

[a] D + 26.4 ° (c 0.10, MeOH)

IR (KBr, m) 3383, 1723 cm ^-1

negative FAB-MS m / z 729 [M ⁻ H] ⁻

GC-MS tR = 11.06 min (palmitic acid)

As a result, compound 7 was prepared as (2S) -1-O-β-D-galactopyranosyl-2,3-di-O-palmitoyl glyceride ((2S) -1-O-β-D-galactopyranosyl- 2,3-di-O-palmitoyl glyceride) was confirmed.

The result of analyzing Compound 8 is as follows.

-Pale yellow wax

[a] D -4.0 ° (c 0.10, MeOH)

IR (KBr, m) 3383, 1723, 1612 cm ^-1

positive FAB-MS m / z 776 [M + H] ⁺

GC-MS t _R = 13.76 min (linolenic acid methyl ester)

As a result, Compound 8 was dissolved in (2S) -1-O-β-D-galactopyranosyl-2,3-di-O-linolenoyl glyceride ((2S) -1-O-β-D-galactopyranosyl -2,3-di-O-linolenoyl glyceride) was confirmed.

The result of analyzing Compound 9 is as follows.

-Pale yellow wax

[a] D + 49.1 ° (c 0.10, MeOH)

IR (KBr, m) 3383, 1723 cm ^-1

positive FAB-MS m / z 655 [M + H] ⁺

GC-MS t _R = 10.27 min (palmitic acid methyl ester)

As a result, Compound 9 was prepared as (2S) -1-O-6'-O- (α-D-galactopyranosyl) -β-D-galactopyranosyl-3-O-palmitoyl glyceride ((2S). ) -1-O-6'-O- (α-D-galactopyranosyl) -β-D-galactopyranosyl-3-Opalmitoyl Glyceride).

The result of analyzing compound 10 is as follows.

-Pale yellow wax

[a] D + 14.4 ° (c 0.10, MeOH)

IR (KBr, m) 3386, 1722, 1610 cm ^-1

positive FAB-MS m / z 677 [M + H] ⁺

GC-MS t _R = 13.76 min (linilenic acid methyl ester)

As a result, Compound 10 was prepared as (2S) -1-O-6'-O- (α-D-galactopyranosyl) -β-D-galactopyranosyl-3-O-linolenoyl glyceride (( 2S) -1-O-6'-O- (α-D-galactopyranosyl) -β-D-galactopyranosyl-3-Olinolenoyl Glyceride).

The analysis of Compound 11 was as follows.

-Pale yellow wax

-[a] D -0.8 ° (c 0.10, MeOH)

IR (KBr, m) 3383, 1723 cm ^-1

Positive FAB-MS m / z 893 [M + H] ⁺

GC-MS t _R = 10.27 min (palmitic acid methyl ester), 11.06 min (palmitic acid)

As a result, Compound 11 was dissolved in (2S) -1-O-6'-O- (α-D-galactopyranosyl) -β-D-galactopyranosyl-2,3-di-O-palmitoyl glycerol. Ride ((2S) -1-O-6'-O- (α-D-galactopyranosyl) -β-D-galactopyranosyl-2,3-di-O-palmitoyl glyceride) was confirmed.

The result of analyzing Compound 12 is as follows.

-Pale yellow wax

[a] D + 7.0 ° (c 0.10, MeOH)

IR (KBr, m) 3386, 1723, 1612 cm ^-1

negative FAB-MS m / z 815 [M + Cl] ^-

GC-MS t _R = 13.76 min (linilenic acid methyl ester), 14.36 min (stearic acid methyl ester)

As a result, compound 12 was dissolved in (2S) -1-O- (6-O-α-D-galactopyranosyl) -β-D-galactopyranosyl-2-O-stearoyl-3-O- Linolenoyl glyceride ((2S) -1-O- (6-O-α-D-galactopyranosyl) -β-D-galactopyranosyl-2-Ostearolyl-3-O-linolenoyl glyc eride) was confirmed.

The result of analyzing Compound 13 is as follows.

-Pale yellow wax

[a] D + 36.6 ° (c 0.10, MeOH)

IR (KBr, m) 3389, 1722, 1615 cm ^-1

Negative FAB-MS m / z 935 [MH] ^-

GC-MS t _R = 13.76 min (linolenic acid methyl ester), 14.58 min (linolenic acid)

As a result of the analysis, compound 13 is (2S) -1-O- (6-O-α-D-galactopyranosyl) -β-D-galactopyranosyl-2,3-di-O-linolenoyl- Glyceride ((2S) -1-O- (6-O-α-D-galactopyranosyl) -β-D-galactopyranosyl-2,3-di-O-linolenoyl-glyceride) was confirmed.

Example 4 GC-MS Analysis of Fatty Acids in Glycerol Glyceride

Each compound (1.0 mg) was dissolved in 2 ml of 20% potassium hydroxide (KOH) / methanol (MeOH) and heated in a water bath at 80 ° C. for 60 minutes. After cooling to room temperature, the reaction mixture was evaluated for disappearance of starting material on TLC plate (n-hexane-EtOAc = 1: 1), then neutralized by addition of acidic cation-exchange resin (Dowex 50W, H ⁺ form) Filtered. Each filtrate was evaporated in vacuo and partitioned between ethyl acetate (EtOAc, 3 ml) and water (3 ml) fractions. The ethyl acetate fraction was then evaporated in vacuo, dissolved in 200 μl of ethyl acetate and filtered through a syringe filter (0.2 μm, 13 mm). The filtrate was stored at -4 ° C until GC-MS analysis.

A DB-5 column (0.25 μm film thickness × 0.25mm diameter, 30m length) was used for the GC-MS experiment. Helium gas was used as the carrier gas at a flow rate of 24.2 ml / min. The oven temperature was programmed to increase at a rate of 4 ° C./min from 160 ° C. to 320 ° C. and held for 15 minutes. The injector and detector temperatures were set to 280 ° C and 280 ° C, respectively. Sample solution (1 μl) was injected into the GC column at a split ratio of 1: 1. Detection was performed by electron ionization (70 eV) and quadrupole mass spectrometry. Fatty acids were identified by retention time and actual fatty acid and mass spectra compared to the library (Wiley Library, version 2008, John Wiley & Sons Inc., Hoboken, NJ, USA).

Example 5: Anticancer Effect Analysis of Mallow Fraction and Compounds 1-13

5-1: Cell Culture

HepG2 (human liver cancer cells), AGS (human gastric cancer cells), HCT-15 (human colon cancer cells) and A549 (human lung cancer cells) cell lines were obtained from the Korean Cell Line Bank (KCLB), and 10% heat-inactivated fetal fetuses. 37 ° C., 5% CO in DMEM and RPMI 1640 supplemented with serum and 1% penicillintreptomycin₂ Incubated in the atmosphere.

5-2: Cell Viability Assay-Confirmation of Apoptosis Effects on Cancer Cells

In order to confirm cytotoxicity against malignant fractions prepared in Examples 2 and 3 and various cancer cells of compounds 1 to 13, the respective fractions and isolated compounds 1 to 13 were applied to HepG2, AGS, HCT-15 and A549 cell lines. Treatment at different concentrations and MTT analysis was performed.

Each cancer cell line was seeded in 100 μl 5 × 10 ³ cells / well in 96-well cell culture plates. After 24 hours, cells were treated with various concentrations of fractions and compounds and incubated at 37 ° C. under 5% CO ₂ atmosphere for 24 hours. The assay was performed by adding 10 μl of 5 mg / mL MTT reagent to the cells and incubating at 37 ° C. for 3 hours. The supernatant of the culture medium was then removed, 100 ml of DMSO was added to each well, and the plate was shaken for 5 minutes. Absorbance at 550 nm was measured using a multi-mode microplate reader (Tecan, Seoul, South Korea). As a result, as can be seen from Table 2, it was confirmed that significantly reduced the survival rate of four cell lines in all fractions and compounds.

TABLE 2

In particular, the n-butanol fraction, Compounds 1, 2 and 11, showed significant cytotoxicity against AGS (human gastric cancer cells), with respective IC ₅₀ values of 8.2 ± 0.14 μg / ml, 33.7 ± 0.64 μM, 11.1 ± 0.07 μM, and 10.6 ± 0.10 μM. From this, mallow extracts, fractions thereof, or compounds isolated therefrom, specifically n-butanol fractions from mallow, compounds 1, 2 and 11 are cytotoxic to various cancer cells, including AGS, preventing or preventing cancer. It was found to have a therapeutic effect.

5-3: Instrumental Analysis of Apoptosis-Confirmation of Induction of Apoptosis

Annex V binding assay and propidium iodide (PI) to confirm the effect of inducing apoptosis on AGS fractions (n-butanol fraction) and AGS of compounds 1, 2 and 11 prepared in Examples 2 and 3 ) Was investigated using staining.

AGS cells were plated at 2 mL 1 × 10 ⁵ cells / well on 6-well cell culture plates and incubated for 24 hours at 37 ° C. in 5% CO ₂ atmosphere. Cell samples were then treated with various concentrations of n-butanol fraction and compounds 1, 2 and 11. After treatment for 24 hours, cells were harvested and analyzed using the Tali Apoptosis Assay Kit according to the manufacturer's instructions.

As a result, the number of apoptotic cells was confirmed to increase as the concentration of n-butanol fraction, compounds 1, 2 and 11 was increased (FIG. 2). That is, the m-butanol fraction of the mallow, compounds 1, 2 and 11 not only exhibits cytotoxicity against cancer cells including AGS, but also shows effects of inducing apoptosis of cancer cells, preventing or treating cancer. It was found to have an effect.

5-4: Western blotting-level analysis of apoptosis related proteins

It was analyzed whether mallow fractions (n-butanol fraction) and compounds 1, 2 and 11 prepared in Examples 2 and 3 regulate the expression of apoptosis-related proteins (PARP, caspase-3, Bax and Bcl-2). . Specifically, PARP (poly (ADP-ribose) polymerase) is a proteolytic substrate of caspase cleaved at the apoptosis stage, the degradation of PARP is induced by the activation of caspase-3, which leads to cell death. In addition, the ratio of Bax (pro-apoptosis gene) and Bcl2 (anti-apoptosis gene) (Bax / Bcl2) is an important factor in determining the stage of cell death.

AGS cells were placed in a 100φ cell culture dish (1 × 10 ⁶ cells / dish in ⁶ mL) and incubated at 37 ° C. in 5% CO ₂ atmosphere. After 24 hours, cells were treated with various concentrations of compound and incubated for 24 hours. Cells were then harvested and protein extracts were prepared using protein extraction kit (iNtRON Biotechnology, Seoul, Korea). Protein extract (30 μg / μl) was separated by 12% SDS-PAGE. The protein was then transferred to nitro cellulose (NC) membranes at 300 mA for 140 minutes. The delivered membrane was blocked and washed for 2 hours with skim milk in TBS containing 0.05% Tween 20 (TBS-T). Subsequently, the membrane was incubated with specific antibodies (PARP, caspase-3, Bax, Bcl-2 and β-actin, 1: 1000) overnight at 4 ° C, washed, and then incubated with secondary antibody at room temperature. Following the manufacturer's instructions, the target protein bands were visualized with EZ-Western Lumi Pico reagent.

As a result, PARP was dose-dependently degraded in the n-butanol fraction, compounds 1, 2 and 11, thereby reducing caspase-3 expression and caspase-3 degradation in AGS cells. On the other hand, Bax was not affected by n-butanol fractions, compounds 2 and 11, but expression was increased by treatment of Compound 1, and Bcl-2 was expressed by treatment of n-butanol fractions, compounds 1, 2 and 11. Decreased (FIG. 3).

As a result, the treatment of the n-butanol fraction, compounds 1, 2 and 11 induced apoptosis by increasing the Bax / Bcl-2 ratio, thereby inhibiting the growth of AGS cells. In addition, compounds 1 and 2, which are sulfoquinovosyl glyceride compounds, are more effective for HepG2, AGS, HCT-15, and A549 cells than other galactoglycosyl glyceride compounds (compounds 3 to 10, 12, and 13). It was confirmed that it has a high anticancer effect. From this, the m-butanol fraction of the mallow, the compounds 1, 2 and 11 was found to have a prophylactic or therapeutic effect of cancer by inducing apoptosis of various cancer cells including AGS.

5-5: Statistical Analysis

Each experiment was performed at least three times. Results are expressed as mean ± standard error (S.E.M.) of the mean. One-way ANOVA was used to evaluate the data at significance levels of * p <0.05, ** p <0.01, and *** p <0.001.

From the above description, those skilled in the art will appreciate that the present invention can be implemented in other specific forms without changing the technical spirit or essential features. In this regard, it should be understood that the embodiments described above are exemplary in all respects and not limiting. The scope of the present invention should be construed that all changes or modifications derived from the meaning and scope of the following claims and equivalent concepts rather than the detailed description are included in the scope of the present invention.

Claims (14)

Malva verticillata extract, fractions thereof, the compounds represented by the following formula (1) isolated from them or a pharmaceutical composition for cancer prevention or treatment comprising a pharmaceutically acceptable salt of the compound as an active ingredient,
[Formula 1]

In the general formula 1 R ₁ is -H, linoleate leno one (linolenoyl), palmitoyl (palmitoyl) or stearoyl one (stearoyl), and; R ₂ is linolenoyl, palmitoyl or stearoyl; R ₃ is —SO ₃ ^— , —OH or glycosyl.
The pharmaceutical composition of claim 1, wherein the extract is extracted using water, C1 to C4 alcohol, or a mixed solvent thereof.
The pharmaceutical composition of claim 2, wherein the C1 to C4 alcohol is methanol.
The pharmaceutical composition of claim 1, wherein the fraction is an n-butanol fraction.
The compound of claim 1, wherein the compound is (2S) -1-O- (6-deoxy-6-sulfo) -α-D-glucopyranosyl-2-O-linolenoyl-3-O-palmitoyl Glyceride (Compound 1), (2S) -1-O- (6-deoxy-6-sulfo) -α-D-glucopyranosyl-2,3-di-O-linolenoyl glyceride (Compound 2 ), (2S) -1-O-β-D-galactopyranosyl-3-O-palmitoyl glyceride (Compound 3), (2S) -1-O-β-D-galactopyranosyl-3 -O-stearoyl glyceride (Compound 4), (2S) -1-O-β-D-galactopyranosyl-3-O-isostearoyl glyceride (Compound 5), (2S) -1 -O-β-D-galactopyranosyl-3-O-linolenoyl glyceride (Compound 6), (2S) -1-O-β-D-galactopyranosyl-2,3-di-O Palmitoyl Glyceride (Compound 7), (2S) -1-O-β-D-galactopyranosyl-2,3-di-O-linolenoyl glyceride (Compound 8), (2S) -1 -O-6'-O- (α-D-galactopyranosyl) -β-D-galactopyranosyl-3-O-palmitoyl glyceride (Compound 9), (2S) -1-O-6 '-O- (α-D-galactopira Sil) -β-D-galactopyranosyl-3-O-linolenoyl glyceride (compound 10), (2S) -1-O-6'-O- (α-D-galactopyranosyl)- β-D-galactopyranosyl-2,3-di-O-palmitoyl glyceride (Compound 11), (2S) -1-O- (6-O-α-D-galactopyranosyl) -β -D-galactopyranosyl-2-O-stearoyl-3-O-linolenoyl glyceride (Compound 12) and (2S) -1-O- (6-O-α-D-galactopira Nosyl) -β-D-galactopyranosyl-2,3-di-O-linolenoyl-glyceride (Compound 13).
The compound of claim 1, wherein the compound is (2S) -1-O- (6-deoxy-6-sulfo) -α-D-glucopyranosyl-2-O-linolenoyl-3-O-palmitoyl Glycerides (compound 1); (2S) -1-O- (6-deoxy-6-sulfo) -α-D-glucopyranosyl-2,3-di-O-linolenoyl glyceride (Compound 2); Or (2S) -1-O-6'-0- (aD-galactopyranosyl) -β-D-galactopyranosyl-2,3-di-O-palmitoyl glyceride (Compound 11) Phosphorus, pharmaceutical composition.
The pharmaceutical composition of claim 1, wherein the composition exhibits cytotoxicity against cancer cells.
The pharmaceutical composition of claim 7, wherein the cancer is gastric cancer, lung cancer, colon cancer, or liver cancer.
The pharmaceutical composition of any one of claims 1 to 6, wherein the composition induces apoptosis of cancer cells.
Malva verticillata ( Malva verticillata ) extract, a fraction thereof, a compound represented by the following formula (1) isolated from them or a food composition for cancer prevention or improvement comprising a pharmaceutically acceptable salt of the compound as an active ingredient,
[Formula 1]

In the general formula 1 R ₁ is -H, linoleate leno one (linolenoyl), palmitoyl (palmitoyl) or stearoyl one (stearoyl), and; R ₂ is linolenoyl, palmitoyl or stearoyl; R ₃ is —SO ₃ ^— , —OH or glycosyl.
The food composition of claim 10, wherein the fraction is an n-butanol fraction.
The compound of claim 10, wherein the compound is (2S) -1-O- (6-deoxy-6-sulfo) -α-D-glucopyranosyl-2-O-linolenoyl-3-O-palmitoyl Glycerides (compound 1); (2S) -1-O- (6-deoxy-6-sulfo) -α-D-glucopyranosyl-2,3-di-O-linolenoyl glyceride (Compound 2); Or (2S) -1-O-6'-0- (aD-galactopyranosyl) -β-D-galactopyranosyl-2,3-di-O-palmitoyl glyceride (Compound 11) Phosphorus, food composition.
(2S) -1-O-β-D-galactopyranosyl-3-O-isostearoyl glyceride ((2S) -1-O-β-D-galactopyranosyl-3-O having anticancer activity -isostearoyl glyceride, compound 5).
A method for preparing a compound represented by the following Chemical Formula 1, comprising separating the compound represented by Chemical Formula 1 from Malva verticillata extract or fraction,
[Formula 1]

In the general formula 1 R ₁ is -H, linoleate leno one (linolenoyl), palmitoyl (palmitoyl) or stearoyl one (stearoyl), and; R ₂ is linolenoyl, palmitoyl or stearoyl; R ₃ is —SO ₃ ^— , —OH or glycosyl.

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