KR101514839B1 - Novel neolignan from magnolia obovata thunb. and an anti-inflammatory composition containing it - Google Patents

Abstract

The present invention is directed to an anti-inflammatory composition. The present invention also relates to novel compounds isolated from yellow magnolia and an anti-inflammatory composition comprising them.

Description

TECHNICAL FIELD [0001] The present invention relates to a novel neo-lignan derived from yellow moss, and an anti-inflammatory composition containing the same. AND ANTI-INFLAMMATORY COMPOSITION CONTAINING IT}

The present invention relates to novel neo-lignans derived from yellow mosses and an anti-inflammatory composition containing the same.

Hwang Magnolia is a deciduous tree that is found throughout Korea, China and Japan. It grows up to 15 m, has long and narrow oval leaves. Magnolia obovata Thunb is known as "Japanese cucumber tree" or "Japanese big leaved magnolia" in the United States and "Honoki" in Japan. Hwang Magnolia's flower is white or orange with a length of 14-20 cm. In addition, the fruit of Huang Magnolia is a pinkish red, oblong-cylinder follicle 12-20 cm long and 6 cm wide, and each corolla is coated with one or two I have black seeds. These seeds ripen from September to October (Kim TW. The woody plants of Korea in color. Korea: Kyo-Hak Publishing Co., Ltd; 1996: 107).

The stem of Magnolia obovata Thunb is a popular herb medicine and is widely used for the treatment of dyspepsia and suppression of sputum (Shin MK, Chung BS. In 'Hyang Yak Dae Sa Jun', 3rd edition. Sa: 1990: 458, 469-471). It is made from three different plant species, Machilus thunbergii, Magnolia officinalis (Magnoliaceae), also known as a hippopotamus, and Magnolia obovata Thunb. (Magnoliaceae).

Hwang Magnolia is widely used in Korea for the treatment of fever, headache, diarrhea, anxiety and alleviation of asthma. However, phytochemical and pharmacological studies have not yet been conducted on the fruit of Huang Magnolia.

Inflammation is a process involving the activation of monocytes and / or macrophages. Activation of macrophages releases many inflammatory mediators such as cytokines, nitric oxide (NO) or prostaglandins. NO and hyperactivity are associated with tissue injury, septic shock, nerve injury, cell death and necrosis (Wang L, Son HJ, Xu ML, Hu JH, Wang MH, papyrifera. J Korean Soc Appl Biol Chem 2010; 53 (3): 297-303). Thus, inhibiting NO production by inhibiting iNOS expression is a useful strategy for the treatment of a variety of inflammatory diseases. LPS is part of the outer membrane of gram-negative bacteria, which triggers the production of cytokines, inflammatory precursors, such as iNOS and COX-2, in a variety of cell types (Kim HJ, Park TS, Jung MS, the anti-oxidant and anti-inflammatory activities of sarcocarp and calyx of persimmon. J Appl Biol Chem 2011; 54 (2): 71-78). The exposure of mononuclear phagocytes to bacterial LPS involves predation, increased oxidative metabolism, and the synthesis and secretion of monokines, as LPS is widely used as a pre-inflammatory stimulus in test settings. Lt; RTI ID = 0.0 > cell < / RTI >

The present inventors have found that neo-lignans are separated from Huang Magnolia while separating and identifying useful compounds from the fruit of Huang Magnolia and found that they have anti-inflammatory activity, thus completing the present invention.

It is an object of the present invention to provide an anti-inflammatory composition.

It is still another object of the present invention to provide novel compounds and salts thereof.

It is another object of the present invention to provide a food composition comprising the novel compound or a salt thereof.

In order to achieve the above object, the present invention provides an anti-inflammatory composition comprising a compound according to any one of the following formulas (I) to (VIV), or a pharmaceutically acceptable salt thereof:

≪ Formula (I) >

≪ Formula (II) >

(Wherein R < 1 > is H or OH)

≪ Formula (III) >

(Wherein R2 is selected from the group consisting of H, OH, COH, COOH, OCH3 and CH2OCH3)

≪ Formula (IV) >

(Wherein R3, R4, R5 and R6 are each independently H or OH)

≪ Formula (V) >

(Wherein R < 7 > is H or OH).

≪ Formula (VI) >

(Wherein R8 and R9 are each independently H or OH)

The present invention also provides a compound selected from the group consisting of the following formulas (1), (3), (5) and (8), or a pharmaceutically acceptable salt thereof. The present invention also provides a food composition comprising said compound or said salt:

≪ Formula (1) >

≪ Formula (3) >

≪ Formula (5) >

≪ Formula (8) >

The compounds of the present invention and the anti-inflammatory composition are effective for treating inflammation by inhibiting the production of inflammatory mediator NO.

1 shows the ¹ H-NMR spectrum of the compound of formula (8).
Figure 2 shows the ¹³ C-NMR spectrum of the compound of formula (8).
Figure 3 shows the HREIMS spectrum of the compound of formula (8).
Figure 4 shows the ¹ H-NMR spectrum of the compound of formula (5).
Figure 5 shows the ¹³ C-NMR spectrum of the compound of formula (5).
Figure 6 shows the gHMBC spectrum of the compound of formula (5).
Figure 7 shows the HREIMS spectrum of the compound of formula (5).
Figure 8 shows the relationship between ¹ H- ¹ H COZY and gHMBC of the compound of formula (I).
9 shows the ¹ H-NMR spectrum of the compound of formula (1).
10 shows a ¹³ C-NMR spectrum of the compound of the formula (1).
11 shows the gHMBC spectrum of the compound of formula (1).
Figure 12 shows the HREIMS spectrum of the compound of formula (1).
13 shows the ¹ H-NMR spectrum of the compound of formula (3).
14 shows the ¹³ C-NMR spectrum of the compound of formula (3).
15 shows the gHMBC spectrum of the compound of formula (3).
Figure 16 shows the HREIMS spectrum of the compound of formula (3).
17 shows the effect of the compounds of formulas 1 to 10 on NO production in RAW 264.7 cells. The test was repeated three times and the data were expressed as mean ± SD. * P <0.05 compared to the group treated with LPS.

The present invention is directed to an anti-inflammatory composition comprising a compound according to any one of formulas (I) to (VI), or a pharmaceutically acceptable salt thereof:

&Lt; Formula (I) >

&Lt; Formula (II) >

(Wherein R &lt; 1 &gt; is H or OH)

&Lt; Formula (III) >

(Wherein R2 is selected from the group consisting of H, OH, COH, COOH, OCH3 and CH2OCH3)

&Lt; Formula (IV) >

(Wherein R3, R4, R5 and R6 are each independently H or OH)

&Lt; Formula (V) >

(Wherein R &lt; 7 &gt; is H or OH).

&Lt; Formula (VI) >

(Wherein R8 and R9 are each independently H or OH)

The present invention also relates to a compound selected from the group consisting of the following formulas (1), (3), (5) and (8) or a pharmaceutically acceptable salt thereof:

&Lt; Formula (1) >

&Lt; Formula (3) >

&Lt; Formula (5) >

&Lt; Formula (8) >

The present invention also relates to a food composition comprising said compound or a salt thereof.

Hereinafter, the present invention will be described in detail.

Antiinflammatory composition

The anti-inflammatory composition of the present invention comprises a compound according to any one of formulas (I) to (VI), or a pharmaceutically acceptable salt thereof:

&Lt; Formula (I) >

&Lt; Formula (II) >

(Wherein R &lt; 1 &gt; is H or OH)

&Lt; Formula (III) >

(Wherein R2 is selected from the group consisting of H, OH, COH, COOH, OCH3 and CH2OCH3)

&Lt; Formula (IV) >

(Wherein R3, R4, R5 and R6 are each independently H or OH)

&Lt; Formula (V) >

(Wherein R &lt; 7 &gt; is H or OH).

&Lt; Formula < VI) >

(Wherein R8 and R9 are each independently H or OH)

More preferably, the anti-inflammatory composition of the present invention comprises a compound selected from the group consisting of formulas (1) to (10) or a pharmaceutically acceptable salt thereof. Even more preferably, the anti-inflammatory composition of the present invention comprises a compound selected from the group consisting of (1), (3), (5) and (8) or a pharmaceutically acceptable salt thereof.

These compounds may be separated from natural products including yellow magnolia, or synthesized. Preferably, the compounds are separated from natural products, and more preferably they are separated from yellow magnolia.

&Lt; Formula (1) >

&Lt; Formula (2) >

&Lt; Formula (3) >

&Lt; Formula (4) >

&Lt; Formula (5) >

&Lt; Formula (6) >

&Lt; Formula (7) >

&Lt; Formula (8) >

&Lt; Formula (9) >

&Lt; Formula (10) >

The anti-inflammatory composition of the present invention inhibits the production of nitric oxide (NO), inhibits the hyperactivity of NO, and inhibits the progress of inflammation. As a result, the anti-inflammatory composition of the present invention prevents tissue wounds, septic shock, nerve damage, cell death and necrosis.

The anti-inflammatory composition of the present invention may be a pharmaceutical composition for the prevention and treatment of inflammatory diseases, a food composition or a cosmetic composition for the prevention and improvement of inflammatory diseases.

Pharmaceutical composition

The anti-inflammatory composition of the present invention may be a pharmaceutical composition for the prevention and treatment of inflammatory diseases. The inflammatory disease may be selected from the group consisting of dermatitis, allergy, atopy, asthma, conjunctivitis, periodontitis, rhinitis, otitis, sore throat, tonsillitis, pneumonia, gastric ulcer, gastritis, Crohn's disease, colitis, hemorrhoids, ankylosing spondylitis, rheumatic fever, lupus, fibromyalgia ), Rheumatoid arthritis, osteoarthritis, rheumatoid arthritis, periarthritis, tendinitis, hay fever, tendinitis, myositis, hepatitis, cystitis, nephritis, sjogren's syndrome, multiple sclerosis and acute and chronic inflammatory diseases It can be either. In addition, the pharmaceutical composition of the present invention can be used for inflammation relief, skin irritation mitigation, skin soothing, prevention and improvement of atopy, and prevention and treatment of acne.

The pharmaceutical composition of the present invention may contain 0.01 to 99.99 parts by weight, preferably 60 to 98 parts by weight, of the compound of the present invention or the anti-inflammatory composition containing the compound of the present invention per 100 parts by weight of the composition. More preferably, the pharmaceutical composition of the present invention may contain 90 to 95 parts by weight of the compound of the present invention per 100 parts by weight of the composition. However, this can be increased or decreased according to the need of the medicinal person, and it can be appropriately increased or decreased according to the situation such as the diet, nutritional status, disease progression, degree of inflammation. In addition, the pharmaceutical composition of the present invention may be formulated into a suitable form together with a pharmaceutically acceptable carrier. &Quot; Pharmaceutically acceptable &quot; refers to compositions which are physiologically acceptable and which, when administered to humans, do not normally cause allergic reactions such as gastrointestinal disorders, dizziness, or the like.

The pharmaceutical composition of the present invention can be administered orally or parenterally and can be used in the form of a general pharmaceutical preparation. The preferred pharmaceutical preparations are those for oral administration such as tablets, hard or soft capsules, liquids, suspensions, etc. These pharmaceutical preparations can be prepared into conventional pharmaceutically acceptable carriers, for example, excipients such as excipients, Binders, disintegrators, lubricants, solubilizers, suspending agents, preservatives or extenders.

The dosage of the pharmaceutical composition of the present invention may be determined by a specialist depending on various factors such as the condition of the patient, age, sex, and complications, but is generally from 0.1 mg to 10 g, preferably from 10 mg to 5 g &Lt; / RTI &gt; Also, the daily dosage of the pharmaceutical composition per unit dosage form, or a half, 1/3 or 1/4 dose thereof, may be contained, and may be administered 1 to 6 times per day. However, in the case of long-term intake for the purpose of health and hygiene or for the purpose of controlling health, the amount may be less than the above range, and the active ingredient may be used in an amount of more than the above range since there is no problem in terms of safety.

Food composition

The food of the present invention may be a health supplement food, a health functional food, a functional food, and the like, but is not limited thereto and includes the addition of the compound of the present invention to natural foods, processed foods, patient foods, and general food ingredients.

The food composition of the present invention can be used as it is or in combination with other food or food compositions, and can be suitably used according to conventional methods. The amount of the active ingredient to be mixed can be suitably determined according to its intended use (prevention, improvement, or therapeutic treatment). In general, the anti-inflammatory composition or compound of the present invention may be added in an amount of 0.1 to 70 parts by weight, preferably 2 to 50 parts by weight, based on 100 parts by weight of the raw material of the food or beverage in the production of food or beverage. The effective dose of the compound may be used in accordance with the effective dose of the pharmaceutical composition, but may be less than the above range for health and hygiene purposes or long-term intake for health control purposes, It can be used in an amount exceeding the range described above.

There is no particular limitation on the kind of the food. The food composition may be used in the form of tablets, hard or soft capsules, liquids, suspensions, and the like, which may contain conventional excipients, such as excipients in the case of oral preparations, Binders, disintegrators, lubricants, solubilizers, suspending agents, preservatives or extenders.

Examples of the food to which the compound or the antiinflammatory composition of the present invention can be added include meat, sausage, bread, chocolate, candy, snack, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, , Beverages, tea, drinks, alcoholic beverages and vitamin complexes, and other nutrients, but are not limited to these kinds of foods.

Cosmetics  Composition

The anti-inflammatory composition of the present invention may be a cosmetic composition. The cosmetic composition of the present invention has preventive and remedial effects on skin inflammation, alleviation of skin irritation, skin soothing, wrinkle improvement and alleviation, inhibition of skin aging, prevention and improvement of acne, prevention and improvement of atopy.

The cosmetic composition of the present invention may be prepared in any formulations conventionally produced in the art and may be formulated into a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant- containing cleansing oil, powder foundation , An emulsion foundation, a wax foundation and a spray, but is not limited thereto. The cosmetic composition of the present invention may also be formulated as a softening agent, a nutritional lotion, a nutritional cream, a massage cream, an essence, an eye cream, a cleansing cream, a cleansing foam, a cleansing water, a pack, a spray or a powder.

The compounds of formulas (1) to (10)

The present invention relates to compounds of formulas (1) to (10) or pharmaceutically acceptable salts thereof, more preferably to compounds of formulas (1), (3), (5) &Lt; / RTI &gt; or a pharmaceutically acceptable salt thereof. These compounds inhibit the production of nitric oxide (NO), suppress the overactive of NO, inhibit the progress of inflammation, and have anti-inflammatory activity. These compounds may be separated from natural products including yellow magnolia, or synthesized. Preferably, the compounds are separated from natural products, and more preferably they are separated from yellow magnolia.

BRIEF DESCRIPTION OF THE DRAWINGS The advantages and features of the present invention, and the manner of achieving them, will be apparent from and elucidated with reference to the embodiments described hereinafter in conjunction with the accompanying drawings. It should be understood, however, that the invention is not limited to the disclosed embodiments, but is capable of many different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete, To fully disclose the scope of the invention to those skilled in the art, and the invention is only defined by the scope of the claims.

Materials and methods

Reagents and devices

Kiesel gel 60 (63-200 μm, Merck) and LiChroprep RP-18 (40-60 μm, Merck) were used for the column chromatography (cc). Thin layer chromatography (TLC) analysis was carried out using Kiesel gel 60 F254 and a RP-18 F254S plates (Merck), and spot on TLC was analyzed using UV lamp spectroline model ENF-240 C / F (UV lamp Spectroline Model ENF-240 C / F (Spectronics Corporation)), sprayed with 10% sulfuric acid solution and confirmed by heating. Deuterium solvent measurement equipment for nuclear magnetic resonance (NMR) Ltd and Sigma Aldrich Co. Lt; / RTI &gt; RPMI medium 1640, Dulbecco's modified Eagle's medium and penicillin and streptomycin were purchased from GIBCO. FBS was obtained from Hyclone, MTT and DMSO were obtained from Sigma Aldrich Co. Lt; / RTI &gt; NMR spectra were recorded on a 400 MHz FT-NMR spectrometer (Varian). IR spectra were obtained from a Perkin Elmer Spectrum One FT-IR spectrometer. EIMS was recorded on JMSAX-700 (JEOL). Optical rotations were measured using a JASCO P-1010 digital polarimeter (JSACO).

plant

The fruit of Magnolia obovata Thunb was collected in September, 2010 at Kyunghee University (Yongin), Korea, and it was confirmed by professor Seungwoo Lee of Kyunghee University. Sulfuretin (≥95% pure) isolated from Rhus verniciflura was used as a positive control (Lee DS, Jeong GS, Li B, Park H, Kim YC. sulfuretin from Rhus verniciflua Stokes via the induction of heme oxygenase-1 expression in murine macrophages. Int Immunopharmacol 2010; 10: 850-8).

Statistical analysis

Test results were expressed as mean ± SD according to three independent tests. One-way analysis of variance (ANOVA) and Newman-Keuls post hoc test were used to compare treatment groups and treatment concentrations. Statistical analysis was performed using GraphPad Prism software version 3.03 (GraphPad Software Inc).

<Experimental Example 1> Separation of Compound from Yellow Magnolia

11 kg of Hwangsugi (dried matter) was cut and repeatedly extracted 4 times with 40 L of 80% methanol at room temperature for 24 hours, filtered and concentrated in vacuo. 740 g of the obtained methanol extract was successfully fractionated with H2O (3.5 L), ethyl acetate (EtOAc) (3.5 L x 4), and n-butanol (3 L x 3). The concentrated ethyl acetate fraction (MOE, 238 g) was dissolved in SiO2 c.c. (12 cm x 15 cm) and monitored by TLC, n-hexane-ethyl acetate (5: 1 → 2: 1 → 1: 2, 2.8 L each) → CHCl3- 15 fractions (MOE-1 to MOE-15) were obtained by eluting with CHCl3-methanol-H2O (6: 4: 1, 3 L) The fraction MOE-2 [77 g, elution volume / total volume (Ve / Vt) 0.023-0.043] was dissolved in SiO2 c.c. (MOE-2-1 to MOE-2-11) by eluting with n-hexane-ethyl acetate (6: 1 -> 4: &Lt; / RTI &gt; Fraction MOE-2-1 (287 mg, Ve / Vt 0.000-0.078) was dissolved in SiO2 c.c. (MOE-2-1-1 to MOE-2-1-14) and (MOE-2-1-1) were obtained by eluting with ethyl acetate (80: 1, 2.0 L) (MOE-2-1-12, 14.4 mg, Ve / Vt 0.528-0.921, TLC (RP-18 F254s) Rf 0.66, methanol-H2O (14: 1)].

The fraction MOE-2-4 [148 mg, Ve / Vt 0.234-0.277] was dissolved in octadecyl silicagel (ODS) c.c. (3.5 cm x 5 cm) and eluted with methanol-H2O (3: 1, 1.2 L) to give 12 fractions (MOE-2-4-1 to MOE-2-4-12) and ultimately produced (MOE-2-4-4, 29.6 mg, Ve / Vt 0.142-0.158, TLC (RP-18 F254s) Rf 0.56, methanol-H2O (7: 1) 2-4-9, 6.6 mg, Ve / Vt 0.226-0.258, TLC (RP-18 F254s) Rf 0.48, methanol-H2O (7: 1)].

Fraction MOE-2-8 [1.02 g, Ve / Vt 0.433-0.543] was dissolved in ODS c.c. (MOE-2-8-1 to MOE-2-8-10) by eluting with acetone-H2O (3: 2 to 3: 1, 1.2 L each) &Lt; / RTI &gt; The fraction MOE-2-8-1 [460 mg, Ve / Vt 0.000-0.042] was dissolved in ODS c.c. (MOE-2-8-1 to MOE-2-8-10) was eluted with acetone-ethanol-H2O (1: 1: 1, 0.6 L) (2: 1: 1) of compound 7 (MOE-2-8-1-5, 69.5 mg, Ve / Vt 0.108-0.127, TLC (RP- 18 F254s) Rf 0.39, acetone-MeOH- )].

Fraction MOE-3 [8.0 g, Ve / Vt 0.057-0.073] was added to SiO2 c.c. (MOE-3-1 to MOE-3-14) was obtained by eluting with ethyl acetate (11: 1, 8.4 L) into a fraction (5.5 cm x 15 cm) Fraction MOE-3-12 [232 mg, Ve / Vt 0.716-0.905] was dissolved in ODS c.c. (3 cm x 5 cm) and eluted with methanol-H 2 O (1: 1, 1.6 L) to give 20 fractions (MOE-3-12-1 to MOE-3-12-20) To obtain compound [MOE-3-12-6, 12.7 mg, Ve / Vt 0.033-0.047, TLC (RP-18 F254s) Rf 0.74, MeOH-H2O (5: 1)].

Fraction MOE-5 [5.3 g, Ve / Vt 0.195-0.286] was added to SiO2 c.c. (5 cm x 13.5 cm) and eluted with CHCl 3 -methanol (40: 1, 4.1 L) to give 7 fractions (MOE-5-1 to MOE-5-7). Fraction MOE-5-2 [1.3 g, Ve / Vt 0.066-0.105] was dissolved in ODS c.c. (4.5 cm x 6 cm) and eluted with acetone-H 2 O (3: 2 → 2: 1 → 5: 2, 2.9 L, respectively) to give 23 fractions (MOE-5-2-1 to MOE- 2-23). The fraction MOE-5-2-3 [73 mg, Ve / Vt 0.039-0.045] was dissolved in ODS c.c. (MOE-5-2-3-1 to MOE-5-2-3-8) was obtained by eluting with methanol-H2O (5: 2, 0.7 L) . The fraction MOE-5-2-3-5 [31 mg, Ve / Vt 0.175-0.219] was dissolved in SiO2 c.c. (2 cm x 13 cm) and eluted with n-hexane-CHCl 3 -methanol (20: 10: 1, 0.6 L) to give 8 fractions (MOE-5-2-3-5-1 to MOE-5 (MOE-5-2-3-5-2, 15 mg, Ve / Vt 0.115-0.128, TLC (RP-18 F254s) Rf 0.60 , Methanol-H2O (8: 1)].

Fraction MOE-5-3 [3.6 g, Ve / Vt 0.105-0.297] was added to SiO2 c.c. (5 cm x 14 cm) and eluted with n-hexane-ethyl acetate (5: 1, 21 L) to obtain 16 fractions (MOE-5-3-1 to MOE-5-3-16) (RP-18 F254s) Rf 0.47, MeOH-H2O (8: 1)].

The fraction MOE-5-3-12 [220 mg, Ve / Vt 0.214-0.257] was dissolved in ODS c.c. (3 cm x 5 cm) and eluted with methanol-H 2 O (3: 2, 5 L) to give 29 fractions (MOE-5-3-12-1 to MOE-5-3-12-29) (RP-18 F254s) Rf 0.64, MeOH-H2O (7: 1)] was finally obtained as the compound [MOE-5-3-12-4, 7.8 mg, Ve / Vt 0.214-0.257, TLC .

The fraction MOE-6 [7.6 g, Ve / Vt 0.287-0.450] was added to SiO2 c.c. (6 cm x 13 cm) and eluted with n-hexane-CHCl 3 -methanol (40: 10: 1, 21.8 L) to obtain eight fractions (MOE-6-1 to MOE-6-8). The fraction MOE-6-5 [665 mg, Ve / Vt 0.242-0.432] was dissolved in ODS c.c. (MOE-6-5-1 to MOE-6-5-) were dissolved in methanol-H2O (1: 1 → 2: 1 → 4: 29-) and ultimately the compound of formula 2 (MOE-6-5-11, 14 mg, Ve / Vt 0.320-0.362, TLC (RP-18 F254s) Rf 0.58, MeOH- .

The fraction MOE-6-5-7 [81 mg, Ve / Vt 0.267-0.307] was added to ODS cc (2.5 cm x 6.5 cm) and eluted with acetone-methanol-H2O (1: 1: 2, 0.8 L) 12 fractions (MOE-6-5-7-1 to MOE-6-5-7-12) were obtained and ultimately the compound of formula 4 [MOE-6-5-7-5, 24.7 mg, Ve / Vt 0.323-0.386, TLC (RP-18 F254s) Rf 0.36, acetone-MeOH-H2O (2: 1: 1)].

<Experimental Example 2> Analysis of Compound

Spectral analysis was performed on the compounds of the formulas (1) to (10) obtained in Experimental Example 1 using NMR, MS and the like.

<2-1> Isobovatol

The compound of formula 8 is isolated as a colorless oil and shows IR absorption at 3368 and 1605 cm-1, indicating the presence of hydroxyl groups and olefin groups. Molecular weights and molecular formulas were identified as 282 and C18H18O3, respectively, from the molecular ion peak [M] + (calculated as C18H18O3, 282.1256) at m / z 282.1254 in HREIMS. The 1 H-NMR spectrum of Compound 3 shows that the 1,2,4-trisubstituted benzene ring protons signal (δH 6.78, 2H, d, J = 8.4 Hz, H-5, 5 ' (隆 H 5.93, 2H, m), J = 1.2 Hz, H-2, 2 'and 隆 H 6.61, 2H, dd, J = 8.4, 1.2 Hz, H- , H-8, 8 ') and exomethylene proton signals (δH 5.05, 2H, d, J = 10.8, 1.2 Hz, H-9a, 9a'; δH 5.03, 2H, d, J = Hz, H-9b, 9b ') and allyl methylene proton signals (δH 3.25, 4H, d, J = 6.8 Hz, H-7, 7'). The NMR data and the molecular ion peak (m / z 282) described above indicate that the compound of formula 8 is a symmetric lignan and consists of two identical phenyl propanoid moieties. The 13 C-NMR spectrum shows 18 carbon signals, which are quaternary carbon signals (δc 143.4, C-4, 4 '; δc 141.6, C-3, 3') of four oxygenated olefins, 2 The carbon signals (δc 133.2, C-1, 1 ') and the eight olefin methine carbon signals (δc 137.5, C-8, 8'; δc 120.9, C-6, 6 '; C-2, 2 ';? c 115.3, C-5, 5'), two exomethylene carbon signals (? c 115.5, C-9, 9 '), and two allylmethylene carbon signals (? c 39.4, C-7, 7 '). These 1 H-NMR and 13 C-NMR spectral characteristics are similar to those of magnolol (Formula 6) and honokiol (Formula 7), which are the main compounds of plants in the magnoliaceae family, with exceptions to the position of the hydroxyl group Do. From the gradient heteronuclear multiple bonding connectivity (gHMBC) spectra, the carbon signal (δc 143.4, C-4, 4 ') with two oxygenated olefins is the olefin methane proton signal (δH 6.78, 2H, d, J = 8.4 Hz, H (Δ C 141.6, C-3, 3 ') showed an olefin methine proton signal (δ H 6.78, H-2, , 2H, d, J = 8.4 Hz, H-5, 5 '; δH 6.61, 2H, dd, J = 8.4, 1.2 Hz, H-6, 6' Is located at C-4 and C-4 ', indicating that two molecules of phenylpropanoid are ether bonded

Colorless oil; IR (CHCl3) .nu.max 3368, 3077, 2924, 2900, 1605, 1516, 1444 cm @ -1; HREIMS m / z: 282.1254266.1311 [M] + (calcd for C18H18O3 282.1256); 1 H-NMR (400 MHz, CDCl 3, 隆 H) and 13 C-NMR (100 MHz, CDCl 3, 隆 C) (Table 1).

location δ _c 隆_H One 133.2 - 2  115.7  6.69, d, 1.2 3  141.6 4  143.4 5  115.3  6.78, d, 8.4 6  120.9
6.61, dd, 8.4, 1.2 7 39.4 3.25, d, 6.8 8 137.5 5.93, m 9 115.5 5.05, m One' 133.2 - 2'  115.7  6.69, d, 1.2 3 '  141.6 4'  143.4 5 '  115.3  6.78, d, 8.4 6 '  120.9 6.61, dd, 8.4, 1.2
7 ' 39.4 3.25, d, 6.8 8' 137.5 5.93, m 9 ' 115.5 5.05, m One'' - - 2'' - - -OCH ₃ - - CHO - -

&Lt; 2-2 > 9-Methoxybovanol

The compound of formula 5 is separated into a dark brown oil and shows absorbance at 3323 and 1599 cm-1, indicating the presence of dihydroxyl and olefin groups. The molecular formula was determined as C19H20O4 from the ion peak [M] + in m / z 312.1350 (calculated as C19H20O4, 312.1350) in HREIMS. The NMR data of the compound of formula (5) were similar to those of obovatal (Formula 4) except for the methoxy group (δH 3.32, δc 57.8) and the oxygenated methylene signal (δH 3.99, δc 73.0) instead of the aldehyde signal of the compound of formula It was very similar to them. The coupling constants between the two olefinic methine proton signals (δH 6.36, 1H, d, J = 16.0 Hz, H-7; δH 6.01, 1H, dt, J = 16.0, 6.4 Hz, H- ) Refers to the trans-configuration of a double bond. The oxygenated methylene proton signal (δH 3.99, H-9) from the gHMBC spectrum correlates with the olefin methine carbon signal (δc 137.3, C-8 '; δc 132.1, C-7) and the methoxy carbon signal (δc 57.8) , Which suggests that the oxygenated methylene carbon is C-9. As a result, the structure of the compound of formula (5) is similar to that of 5- (3-methoxy-1-propen-1-yl) -3- [4- (2-propan-1-yl) phenoxy] (2-propene-1-yl) phenoxy] -1,2-benzenediol), which was identified as 9-methoxy Called 9-methoxyobovatol (Figures 4 to 7).

Dark brown oil; IR (CHCl3) .nu.max 3323, 3066, 2925, 2819, 1599, 1505, 1447, 1326, 1217 cm @ -1; HREIMS m / z: 312.1350 [M] + (calcd. For C19H20O4 312.1350); 1 H-NMR (400 MHz, CDCl 3, 隆 H) and 13 C-NMR (100 MHz, CDCl 3, 隆 C) (Table 2).

location δ _c δ _H One 135.4 - 2 108.7 6.44, d, 1.6 3 144.1 - 4 134.5 - 5 144.9 - 6 108.8 6.74, d, 1.2 7 132.1 6.36, d, 16.0 8 124.3 6.01, dt, 16.0, 6.4 9 73.0 3.99, dd, 6.4, 1.2 One' 129.2 - 2' 118.2 6.93, d, 8.4 3 ' 129.9 7.13, d, 8.8 4' 154.7 - 5 ' 129.9 7.13, d, 8.8 6 ' 118.2 6.93, d, 8.4 7 ' 39.4 3.35, d, 6.4 8' 137.3 5.93, m 9 ' 115.9 5.09, m One'' - - 2'' - - -OCH ₃ 57.8 3.32, s CHO - -

<2-3> Magnobobartol

The compound of formula (I) is separated into a brown oil and exhibits IR absorption bands of 3391 and 1601 cm-1, indicating the presence of hydroxyl and olefin groups. The optical coherence showed dextrorotatory characteristics [α] D + 7.40 °. The molecular formula was identified as C21H24O5 from the ion peak [M] + at m / z 356.1621 of HREIMS (calculated as C21H24O5, 356.1624). The 1 H-NMR spectrum shows that the four olefinic methine proton signals (δH 7.07, 2H, d, J = 8.0 Hz, H-2 ', 6'; δH 6.79, 2H due to the para-disubstituted benzene ring , d, J = 8.0 Hz, H-3 ', 5') and the 1,2,3,4,5-pentasubstituted benzene ring One of the aromatic proton signals (s, 1H, H-6) was shown. Two overlapping olefinic methine proton signals (δH 5.96, 2H, m, H-8, 8 ') were also observed. (ΔH 5.05, 4H, m, H-9,9 '), one hemiacetal proton signal (δH 4.38, 1H, t, J = 4.8 Hz, H-2 " Of methoxy proton signals (δH 3.21, 3H, s) were observed. In the high magnetic field, three allylmethylene proton signals (δH 3.38, 2H, d, J = 4.8 Hz, H-7; δH 3.31, 2H, d, J = 6.8 Hz, H- d, J = 4.8 Hz, H-1 &quot;). Taken together, these data indicate that the compound of formula (1) is a neo-lignan having a side chain. The &lt; 13 &gt; C-NMR spectrum showed 21 carbon signals including one methoxy group (delta c 54.1, -OCH3). In the authors 'group, four oxygenated olefin 4 carbon signals (δc 155.5, C-4'; δc 143.3, C-5; δc 140.4, C-3; δc 134.5, C-4) (隆 c 134.1, C-1 '; 隆 c 132.3, C-2; 隆 c 120.8, C-1), seven olefin methine carbon signals -8;? C 129.9, C-2 ', 6';? C 114.6, C-3 ', 5';? C 113.7, C-6 and two exomethylene carbon signals? C 115.7, C-9 ' 115.6, C-9) were observed. (3), the hemiacetal carbon signal (? C 105.4, C-2 ") and three methylene carbon signals (? C 39.2, C-7 ';? C 36.8, C-7;? C 30.7, C-1") were observed. Thus, the compound of formula 1 was identified as a neo lignan with a 1-methoxyethanol moiety. 2D NMR tests such as gCOSY, gHSQC, and gHMBC were performed to determine the position of the functional group. The linkage between C-1 "and C-2" was evidenced from gCOSY cross-peaks (FIG. 8). In the gHMBC spectrum, the methylene proton signal (δH 2.70, H-1 ") is the carbon signal (δc 140.4, C-3) composed of oxygenated olefin 4 groups and the carbon signal (δc 132.3, C- 2;? C 120.8, C-1), indicating that the methylene carbon signal (C-1 ") is linked at the C-2 position. An additional hemiacetal proton (δH 4.38, H-2 ") is associated with the methoxy carbon signal (δc 54.1, -OCH3) indicating that the methoxy group is in the C-2" position. As a result, the compound of formula (I) can be obtained by reacting 2- (2-methoxyethanol) -1- (2-propen- , 2-methoxyethanol-1- (2-propen-1-yl) -5- [4- (2-propen- 1 -yl) phenoxy] -3,4-benzenediol And was named magnobovatol (Figures 9-12).

Brown oil; [?] D + 7.40 (c = 0.060, CHCl3); IR (CHCl3) .nu.max 3391, 3080, 2927, 1601, 1505 cm @ -1; HREIMS m / z: 356.1621 [M] + (calcd for C21H24O5 356.1624); 1 H-NMR (400 MHz, CDCl 3, 隆 H) and 13 C-NMR (100 MHz, CDCl 3, 隆 C) (Table 3).

location δ _c 隆_H One 120.8 - 2 132.3 - 3 140.4 - 4 134.5 - 5 143.3 - 6 113.7 6.67, s 7 36.8 3.38, d, 4.8 8 137.2 5.96, m 9 115.6 5.05, m One' 134.1 - 2' 129.9 7.07, d, 8.0 3 ' 114.6 6.79, d, 8.0 4' 155.5 - 5 ' 114.6 6.79, d, 8.0 6 ' 129.9 7.07, d, 8.0 7 ' 39.2 3.31, d, 6.8 8' 137.4 5.96, m 9 ' 115.7 5.05, m One'' 30.7 2.70, d, 4.8 2'' 105.4 4.38, t, 4.8 -OCH ₃ 54.1 3.21, s CHO - -

<2-4> 2-Hydroxyobaldealdehyde

Compounds of formula 3 are separated into brown oil and show IR absorption band at 3289, 1671, and 1596 cm-1, indicating the presence of hydroxyl, conjugated carbonyl and olefin groups. The molecular formula was determined from the molecular ion peak [M] + in HREIMS at m / z 286.0851 (calculated as C19H20O4, 286.0841), which is 16 units larger than that of obovaaldehyde (Formula 2) Indicates that the compound of formula (3) has one additional hydroxyl group as compared to the compound of formula (2). The NMR signals of the compound of formula (3) can be obtained by the presence of a 1,2,3,4,5-pentasubstituted benzene ring in place of the 1,2,3,5-tetrasubstituted benzene ring of the compound of formula (2) Except for this, it is similar to the compound of formula (2). In the gHMBC spectrum, the aldehyde carbon (δc 190.6) correlated with the olefinic methine proton signal (δH 6.92, s, H-6), indicating that the aldehyde is in the C-1 position. The olefinic methine proton of H-6 is correlated with the carbon signal (δ c 145.1 C-2, 4) of the oxygenated olefin 4 group. The structure of the compound of formula (3) can be converted to the bisnorneolignan 1-methanol-5- [4- (2-propen-1-yl) phenoxy] -2,3,4-benzenetriol 1-methanol-5- [4- (2-propen-1-yl) phenoxy] -2,3,4-benzenetriol) was named as 2-hydroxyobovaaldehyde) 13 to 16).

Dark brown oil; IR (CHCl3) .nu.max 3289, 3075, 2890, 2725, 1671, 1596, 1505 cm @ -1; HREIMS m / z: 286.0851 [M] + (calcd. For C16H14O5 286.0841); 1 H-NMR (400 MHz, CDCl 3, 隆 H) and 13 C-NMR (100 MHz, CDCl 3, 隆 C) (Table 4).

location δ _c δ _H One 111.2 - 2 145.1 - 3 129.1 - 4 145.1 - 5 140.1 - 6 110.7 6.92, s 7 - - 8 - - 9 - - One' 136.5 - 2' 130.1 6.97, d, 8.0 3 ' 118.9 7.18, d, 8.0 4' 153.7 - 5 ' 118.9 7.18, d, 8.0 6 ' 130.1 6.97, d, 8.0 7 ' 39.4 3.38, d, 6.8 8' 137.0 5.99, m 9 ' 116.1 5.10, m One'' - - 2'' - - -OCH ₃ - - CHO 190.6 9.66, s

<2-5>

NMR and MS data showed that the compound of formula 2 was identified as obovaaldehyde and the compound of formula 4 was identified as obovata. The compound of formula (6) was identified as magnolol, the compound of formula (7) was identified as honokiol, the compound of formula (9) was identified as isomagnolol, It was identified as obovatol.

&Lt; Experimental Example 3 > Cell survival rate and cytotoxicity

RAW 264.7 macrophages (1 x 104 cells / well) were cultured in 96-well plates for 12-18 hours and incubated at various concentrations of neo-lignans diluted in serum-free medium Lt; / RTI &gt; (1 &lt; RTI ID = 0.0 &gt; pg / mL) &lt; / RTI &gt; Thereafter, 3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide (3- (4,5- MTT) reagent (Sigma-Aldrich Co.) was added to the wells and incubated for 4 hours. The formazan formed in the cell pellet was dissolved by adding 100 μL of DMSO (Sigma-Aldrich Co.), and the absorbance was measured at 550 nm. The cell viability was calculated as a percentage of viable cells by the following formula 1.

<Formula 1>

Cell survival rate (%) = [OD (compound) - OD (blank)} / {OD (control group) - OD (blank)

The cytotoxicity of the compounds of formulas 1 to 10 was also assessed by MTT assay, with no cytotoxic activity against RAW 264.7 macrophages in the range of 5-30 μM. At this time, 40 μM of Sultifuretin was used as a positive control group.

<Experimental Example 4> Evaluation of anti-inflammatory activity

As described in Jeong RH et al., NO was confirmed by measuring the amount of nitrite, which is a stable oxide in the supernatant of cell culture (Jeong RH, Lee DY, Cho JK, Lee SM, Kang HC, Seo WD, Kang HW, Kim JY, Baek NI. A new flavonolignan from the aerial parts of Oyasa stiva L. inhibits nitric oxide production in RAW 264.7 macrophage cells. J Korean Soc Appl Biol Chem 2011; 54 (6): 865-870. RAW 264.7 cells (1 x 104 cells / well) were cultured in 96 well cell culture plates for 12-18 hours, and then incubated for 24 hours with compound Were treated with 1 μg / mL LPS in serum-free medium containing various concentrations (5, 10, 15, 20 μM). 100 μL of cell culture supernatant was then mixed with 100 μL of Griess reagent (Sigma-Aldrich Co.) in a new 96-well plate and absorbance was measured at 550 nm using a spectrophotometer. Nitrite concentrations were determined by comparison with the sodium nitrite standard curve.

As a result, the compounds of formulas (1), (3) and (5) to (10) excellently suppressed LPS-induced NO production in proportion to the treated concentration of 15, 30 μM (FIG. These results were obtained by repeating the experiment 3 times. The IC50 values for the NO production measured based on this were 14.2 ± 1.2 μM in the formula 1 and 14.8 ± 3.2 μM in the compound of the formula 3, respectively. On the other hand, the IC50 values of the compounds of Formulas 5 to 10 were 14.8 ± 2.3, 15.8 ± 0.3, 3.3 ± 1.2, 12.9 ± 2.0, 14.1 ± 0.9 and 6.2 ± 1.2 μM, respectively.

Claims (11)

An antiinflammatory composition comprising a compound according to any one of the formulas selected from the group consisting of the following formulas (1), (3), (5), (8) and (9), or a pharmaceutically acceptable salt thereof:

&Lt; Formula (1) >

&Lt; Formula (3) >

&Lt; Formula (5) >

&Lt; Formula (8) >

&Lt; Formula (9) >

delete The method according to claim 1,
Wherein the composition inhibits over-activity of nitric oxide.
The method according to claim 1,
Wherein the composition is a pharmaceutical composition for the prophylaxis and treatment of inflammatory diseases.
The method according to claim 1,
Wherein the composition is a food composition for preventing and ameliorating an inflammatory disease.
The method according to claim 1,
Wherein the compound is separated from the yellow magnolia.
A compound selected from the group consisting of the following formulas (1), (3) and (8) or a pharmaceutically acceptable salt thereof:

&Lt; Formula (1) >

&Lt; Formula (3) >

&Lt; Formula (8) >

8. The method of claim 7,
Lt; RTI ID = 0.0 &gt; anti-inflammatory &lt; / RTI &gt; activity.
8. The method of claim 7,
Lt; RTI ID = 0.0 &gt; nitric oxide. &Lt; / RTI &gt;
8. The method of claim 7,
Sulfur &lt; / RTI &gt; magnolia.
A food composition comprising the compound of claim 7.

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