KR100263439B1 - Novel lignan compounds, process for preparation from magnoliae flos and use for antagonist of platelet-activating factor receptor - Google Patents

Abstract

PURPOSE: A lignan compound separated from Magnoliae flos is provided, which is useful in manufacturing platelet-activating factor and preventing and treating inflammatory diseases. And its manufacturing method is also provided. CONSTITUTION: The method for separating compounds of the formula (1) and (2) is comprised of the next steps of: i) mashing 3kg of Magnoliae flos and obtaining extracts using methanol; ii) condensing the extracts; iii) suspending the extracts in distilled water and obtaining ethylacetate; iv) obtaining the two compounds from ethylacetate by silica gel column chromatography. The compound of the formula (1) has no color, and molecular weight of 402. And its molecular formula is C22H26O7. The compound of the formula (2) has no color and molecular weight of 432 and its molecular formula is C23H28O8.

Description

New Lignan Compounds Isolated from Sinai, Methods for their Preparation, and Antagonists Acting on Platelet Activation Factor Receptors Having the Same As Active Ingredients

The present invention relates to novel lignan compounds represented by formulas (1) and (2), respectively named Magnon A and Magnon B.

Formula 1

Formula 2

The present invention also relates to a process for preparing the compounds of the formulas (1) and (2) from Magnoliae flos.

In addition, the present invention relates to an antagonist that acts on platelet-activating factor (PAF) receptors comprising the new compounds of formulas (1) and (2) as an active ingredient, which prevents inflammatory diseases associated with the platelet activating factor and It can be usefully used for treatment.

The present invention also relates to an antagonist which acts on the extract and the platelet activating factor receptor having the same as an active ingredient.

Magnoliae flos is a dried , unburned bud of the Magnoliae forgesii Cheng ( M. biondii Pamp.) Belonging to the Magnoliaceae, with a spicy taste (Namba, T., 1980, Colored Ilustration of Wakan-Yaku: Hoikusha Publishing, Osaka, Vol. II, 127-129). It has been used to treat headaches, malignant mucosal ulcers, sinusitis, and toothache because of its sensation of swelling and stagnation from ancient times. , Springer-Verlag, Berlin, 639-646).

To date, various kinds of essential oils, lignans, sesquiterpenes, etc. have been separated from the gods, and they have been reported to exhibit calcium (Ca ²⁺ ) antagonistic activity, platelet activating factor (PAF) antagonistic activity and antihistamine activity (Jung, KY et al., 1997, Arch. Pharm. Res., 20, 363-367; Chen, CC et al., 1988, Planta Med., 54, 438-440; Pan, JX et al., 1987, Phytochemistry, 26, 1377-1379; Nagashima, S. et al., 1981, J. Takeda Res. Lab., 40, 27-36).

Platelet activating factor (PAF) is a factor that activates platelets by stimulating the basophils of rabbits sensitized with immunoglobulin E (IgE) to the culture medium, activating platelets. Produced in immune cells such as phagocytes, mast cells, and vascular endothelial cells and platelets (Benveniste, J. et al., 1972, J. Exp. Med., 1356-1377; Benveniste, J. and B. Arnoux, 1983, Platelet-activating factor and structurally related ether-lipids, Elsevier, Amsterdam; Benveniste, J. and BB Vargaftig, 1983, Platelet-activating factor, Academic Press, New York, 355-476; Lee, TC and F. Snyder, 1984, Function metabolism and regulation of platelet-activating factor and related ether lipids in phospholipids and cellular regulation, CRC Press, F1, 1-39).

In addition, platelet activating factors are 1 / 1,000-1 / 10,000 lower (10 ^{-9-10 -10} M) than physiologically inflammatory mediators such as histamine, leukotriene and prostaglandin. Can activate inflammatory cells and act as initiators of acute inflammatory responses leading to the synthesis and release of other inflammatory mediators (Braquet, P. et al., 1987, Pharmacol. Rev., 39, 97- 145). Therefore, when the inflammatory response acting by platelet activator is activated, various inflammatory diseases such as bronchial contraction, asthma, tissue transplant rejection, cardiac anaphylaxy, endotoxin shock, gastric ulcer, acute allergic reaction, and irritable dermatitis are caused ( Gonzalez-Crussi, F. and W. Hsueh, 1983, Am. J. Pathol., 112, 127-135; Katoh, N. et al., 1994, Nippon Shokakibyo Gakkai Zasshi, 91, 27-35; Frerichs, KU et al., 1990, J. Neurosurg., 73, 223-233; Tokutomi, T. et al., 1994, Acta. Neurochir.Suppl.Wien., 60, 508-510; Makowka, L. et al., 1990, Transplantation, 50, 359-365).

However, if platelet activating factor (PAF) is not bound to the receptors, the inflammatory disease is improved by inactivating lyso-platelet activating factor (lyso-PAF) by the hydrolase acetylhydrolase in plasma or cells. For this purpose, a search has been made for substances that antagonistically inhibit the binding of platelet activating factors to platelet membrane receptors (Braquet, P. et al., 1987, Pharmacol. Rev., 39, 97-145).

The antagonists having a structure similar to platelet activators have been reported to CV 3988, CV 6909, ONO 6240, SRI series, and platelet activator antagonists derived from natural products from ginkgo biloba ginkgolide B, etc. Kadsurenone from vines, coumarin from white flower buds, and magnolin from gods have been reported (Terashita, ZI et al., 1983, Life Sci., 32, 1975-1982; Braquet , P., 1985, Prostaglandin, 30, 689; Shen, TY et al., 1985, Proc. Natl. Acad. Sci. USA, 82, 672-676; Takeuchi, N, et al., 1988, Chem. Pharm Bull., 36, 4221-4224; Pan, JX et al., 1987, Phytochemistry, 26, 1377-1379).

Accordingly, the present inventors have identified a substance having an antagonistic activity that acts on platelet activator receptors from the gods to develop a therapeutic agent for inflammatory diseases related to platelet activator receptors from traditional natural plants, and from these two new lignan compounds The present invention was completed by investigating the antagonistic activity of inhibiting the action of platelet activating factor receptor using the compound and investigating its structure by spectroscopic method.

It is an object of the present invention to provide a novel lignan compound, a method for preparing them from gods, and an antagonist acting on platelet activating factor receptors having these as active ingredients and god extracts.

Figure 1 shows the hydrogen nuclear magnetic resonance spectrum results of the compound of formula 1 of the present invention,

Figure 2 shows the carbon nuclear magnetic resonance spectrum results of the compound of formula 1 of the present invention,

Figure 3 shows the hydrogen nuclear magnetic resonance spectrum results of the compound of formula 2 of the present invention,

Figure 4 shows the carbon nuclear magnetic resonance spectrum results of the compound of formula 2 of the present invention.

In order to achieve the above object, the present invention provides novel lignan compounds of the formulas (1) and (2), their derivatives and their salts.

In another aspect, the present invention provides a method for producing the compound by extracting the thin alcohol with lower alcohol, suspended with distilled water, washed with hexane and then extracted again with ethyl acetate to repeat the adsorption chromatography.

The present invention also provides a pharmaceutical composition for antagonists that act on platelet activating factor receptors comprising the compounds of formula 1 and / or formula 2 and derivatives thereof as an active ingredient.

Antagonists acting on the platelet activating factor receptor may be used as a prophylactic, therapeutic and therapeutic adjuvant for inflammatory diseases associated with platelet activating factors or as a modulator of pregnancy and egg implantation.

In addition, the present invention provides a shinto extract having an antagonistic activity against the platelet activating factor receptor comprising a compound of formula (1) and a compound of formula (2) obtained by extracting sinyi with a lower alcohol and an organic solvent.

Hereinafter, the present invention will be described in detail.

The present invention has discovered antagonistic activity from Magnoliae flos in the process of obtaining a new substance exhibiting antagonistic activity by acting on platelet activator receptors from traditional natural plants.

The present invention provides a novel lignan compound, derivatives thereof and salts thereof represented by formulas (1) and (2) by extracting sini with lower alcohol.

Formula 1

Formula 2

Specifically, Sinyi is extracted with lower alcohol, filtered, concentrated under reduced pressure, suspended with distilled water, washed with hexane, extracted again with ethyl acetate, and repeated adsorption chromatography to prepare the compounds of Chemical Formulas 1 and 2.

In this case, the lower alcohol is preferably methanol, and the adsorption chromatography is preferably silica gel column chromatography or silica gel reverse phase column chromatography. In this process, the elution solvent is chloroform: methanol mixed solvent, methanol: water mixed solvent, and the like. Preference is given to using.

The present invention also provides a pharmaceutical composition comprising the compounds of formula 1 and / or formula 2 and their derivatives as active ingredients as antagonists acting on platelet activating factor receptors.

Antagonists acting on the platelet activator receptor may be usefully used as a prophylactic, therapeutic and therapeutic adjuvant for inflammatory diseases associated with platelet activating factors or as a modulator of pregnancy and egg implantation. Specifically, the inflammatory diseases include asthma, allergies, arthritis, rheumatism, thrombosis, acute inflammation, systemic hypersensitivity, retinitis, hepatitis, enteritis, pancreatitis, nephritis, rhinitis, atopic dermatitis, heart hypersensitivity, endotoxin shock, central nervous system damage or Organ transplant rejection.

Pharmaceutical compositions for antagonists of the platelet activating factor of the present invention may be prepared in unit dose form, in multi-dose form, or by incorporation into a multi-dose container by formulating with a pharmaceutically acceptable carrier, excipient. have.

The preparation form can then be in the form of solutions, suspensions or emulsions in oils or aqueous media or in the form of extracts, powders, granules, tablets or capsules, and can include dispersants, suspensions or stabilizers.

In another aspect, the present invention provides a shintoise extract having an antagonistic activity to the platelet activating factor receptor comprising a compound of formula (1) and / or a compound of formula (2) obtained by extracting sinyi with a lower alcohol.

In addition, in addition to the above process, suspended extracts obtained by suspending in distilled water, washed with hexane and extracted again with ethyl acetate.

Hereinafter, the present invention will be described in detail by way of examples.

The following examples illustrate the invention and are not intended to limit the scope of the invention.

<Example 1> Isolation of the Compounds of Formula 1 and Formula 2 from Sinyi

3 kg of Shin was triturated, extracted four times with methanol at room temperature, and the filtrate was collected and concentrated under reduced pressure to obtain 225 g of extract. The extract was suspended in distilled water, washed three times sequentially with hexane (n-hexane), and the aqueous layer was extracted three times with ethyl acetate to obtain 109 g of ethyl acetate fraction.

The ethyl acetate fraction was separated into 19 fractions by silica gel column chromatography (eluent, CHCl ₃ -MeOH; 100: 0 → 0: 100 concentration gradient), and the 14th fraction was silica gel reversed phase (RP-18) column chromatography. (Eluent, MeOH-H ₂ O; 2: 1), and the mixture was divided into 11 small fractions. The fifth subfraction was again subjected to silica gel column chromatography (eluent, CHCl ₃ -MeOH; 99: 1) to purely isolate 28 mg of the compound of Formula 1 and 84 mg of the compound of Formula 2.

Example 2 Physicochemical Characteristics of Compounds of Formulas 1 and 2

The physicochemical characteristics of the compound of Formula 1 and the compound of Formula 2 separated from Sinyi are as follows.

(2-1) Compound of formula (1): (2S, 3R, 4R) -tetrahydro-2- (3,4-dimethoxyphenyl) -4- (3,4-dimethoxybenzoyl) -3-furanmethanol

Formula 1

(1) Property of the substance: Colorless powder

(2) Molecular Weight: 402

(3) Molecular formula: C ₂₂ H ₂₆ O ₇

(4) hydrogen nuclear magnetic resonance spectrum:

Hydrogen nuclear magnetic resonance spectra measured using chloroform-d (CDCl ₃ ) as a solvent and tetramethyl silane (TMS) as a standard are as follows (see FIG. 1 ).

(Chloroform-d, δ) -2.85 (1H, m), 3.63 (1H, dd, J = 10.9, 5.6 Hz), 3.72 (1H, dd, J = 10.9, 4.4 Hz), 3.81 (3H, s), 3.85 (3H, s), 3.88 (3H, s), 3.89 (3H, s), 4.13 (2H, m), 4.25 (1H, dd, 11.2, 10.7 Hz), 4.63 (1H, d, J = 9.1 Hz ), 6.76 (1H, d, J = 8.3 Hz), 6.86 (1H, d, J = 8.3 Hz), 6.88 (1H, dd, J = 8.3, 1.9 Hz), 6.96 (1H, d, J = 1.9 Hz ), 7.51 (1H, d, J = 2.0 Hz), 7.54 (1H, dd, J = 8.3, 2.0 Hz)

(5) Carbon Nuclear Magnetic Resonance Spectrum:

The results of carbon nuclear magnetic resonance spectra measured using chloroform-d (CDCl ₃ ) as a solvent and tetramethyl silane (TMS) as a standard (see FIG. 2 ) are as follows.

(Chloroform-d, δ)-49.66, 52.09, 55.88, 56.11, 61.43, 70.82, 83.76, 109.54, 110.08, 110.55, 110.79, 119.30, 123.16, 129,69, 132.88, 148.89, 149. 20, 153.65, 198.01

(2-2) A compound of formula 2: (2S, 3R, 4R) -tetrahydro-2- (3,4,5-dimethoxyphenyl) -4- (3,4-dimethoxybenzoyl) -3-furan Methanol

Formula 2

(1) Appearance of substance: Colorless oil

(2) Molecular weight: 432

(3) Molecular formula: C ₂₃ H ₂₈ O ₈

(4) hydrogen nuclear magnetic resonance spectrum:

Hydrogen nuclear magnetic resonance spectra measured using chloroform-d (CDCl ₃ ) as a solvent and tetramethyl silane (TMS) as a standard are as follows (see FIG. 3 ).

(Chloroform-d, δ) -2.85 (1H, m), 3.65 (1H, dd, J = 11.0, 4.5 Hz), 3.72 (1H, dd, J = 11.0, 4.5 Hz), 3.75 (3H, s), 3.82 (6H, s), 3.87 (3H, s), 3.89 (3H, s), 4.11 (1H, m), 4.15 (1H, m), 4.23 (1H, t, J = 11.1 Hz), 4.60 (1H , d, J = 8.8 Hz), 6.60 (2H, s), 6.80 (1H, d, J = 8.3 Hz), 7.50 (1H, d, J = 2.0 Hz), 7.54 (1H, dd, J = 8.3, 2.0 Hz)

(5) Carbon Nuclear Magnetic Resonance Spectrum:

The results of carbon nuclear magnetic resonance spectra measured using chloroform-d (CDCl ₃ ) as a solvent and tetramethyl silane (TMS) as a standard (see FIG. 4 ) are as follows.

(Chloroform-d, δ)-49.56, 52.13, 55.96, 56.10, 61.39, 70.88, 83.86, 103.56, 110.06, 110.52, 123.14, 129.64, 136.24, 137.59, 149.18, 153.29, 153.64, 197.75

<Example 2> Antagonistic activity test acting on platelet activating factor receptor

In order to investigate the antagonistic activity of the compounds of Formula 1 and the compounds of Formula 2 against platelet activator receptors, methods such as slightly modified balons and the like were used (Valone, FH et al., 1982). , J. Immunol., 129, 1637-1641; Yang, HO et al., 1995, Planta Med., 61, 37-40).

Specifically, 100 μl of a rabbit platelet suspension (4 × 10 ⁸ cells / ml) was added to 60 μl of a sample solution at a constant concentration, and allowed to stand at room temperature for 10 minutes. The reaction mixture was stopped by filtration under reduced pressure with a membrane filter pre-soaked with Tris buffer and dried at room temperature. The dried membrane filter was placed in each vial, and then 3 ml of radioactive cocktail solution was added to measure radioactivity with a liquid scintillation counter. Antagonistic activity for each sample was calculated by the following equation 1 as the average value of three samples.

Antagonistic activity (%) = (Sc-Ss) / Sc × 100

= [(Tc-Nc)-(Ts-Ns)] / (Tc-Nc) × 100

In Equation 1,

Sc: specific binding of the control group,

Tc: total binding of the control group,

Nc: nonspecific binding of the control group,

Ss: specific binding of sample group,

Ts: total binding of sample groups,

Ns: nonspecific binding of sample group

The methanol extract and the ethyl acetate fraction of Xinyi were respectively added to measure the phenomenon of inhibiting specific binding of platelet activating factor receptors, and the results are shown in Table 1.

Antagonistic Activity of Platelet Activating Factor Receptor Binding to Methanol Extracts and Their Ethyl Acetate Fraction Concentration (μg / ml) PAF receptor binding inhibition rate (%) Methanol extract 200 99.0 Ethyl acetate fraction 200 95.0

From the results of Table 1, the phenomenon of inhibiting specific binding of the platelet activator receptor by adding the compound of the formula (1) and the compound of the formula (2) obtained by chromatographic analysis of the ethyl acetate fraction having antagonistic activity to the platelet activator receptor It was measured, and the results are shown in Table 2.

Antagonistic Activity of Platelet Activator Factor Receptors of Compounds of Formula 1 and Compound 2 Concentration (μM) Platelet activator receptor binding inhibition rate (%) Compound of Formula 1 Compound of formula 2 100 75.2 81.3 50 56.7 64.3 25 41.6 42.9 12.5 19.4 37.1 IC ₅₀ = 3.8 × 10 ^-5 M IC ₅₀ = 2.7 × 10 ^-5 M

As described above, the present invention provides a antagonist for platelet activating factor receptor isolating the compounds of formula (1) and compounds of formula (2) from the god to the active ingredient, the compound of the present invention and As shown in Table 1 and Table 2 it can be seen that the antagonistic activity against platelet activating factor receptor is very excellent.

Therefore, the antagonist acting on the platelet activator receptor of the present invention can be used to regulate the activity of platelet activator in vivo or ex vivo, specifically inflammatory diseases associated with platelet activator factor, asthma, allergy, arthritis, nephritis, rhinitis, It can be useful for the prevention and treatment of diseases such as atopic dermatitis, cardiac hypersensitivity, endotoxin shock, central nervous system damage or organ (tissue) transplant rejection, and control of pregnancy and egg implantation.

Claims (8)

New lignan compounds represented by the formula (1), derivatives thereof and salts thereof. Formula 1

New lignan compounds represented by the formula (2), derivatives thereof and salts thereof. Formula 2

A method of preparing a compound of Formula 1 and a compound of Formula 2, which is extracted with lower alcohol, suspended with distilled water, washed with hexane, and extracted again with ethyl acetate to repeat adsorption chromatography. A pharmaceutical composition for antagonists that acts on platelet activating factor receptors comprising the compounds of Formula 1 and / or Formula 2 and derivatives thereof as an active ingredient. The pharmaceutical composition of claim 4, wherein the antagonist acting on the platelet activator receptor is a prophylactic, therapeutic and therapeutic adjuvant for inflammatory diseases associated with platelet activating factor or a modulator of pregnancy and egg implantation. The method of claim 5, wherein the inflammatory disease is asthma, allergy, arthritis, rheumatism, thrombosis, acute inflammation, systemic hypersensitivity, retinitis, hepatitis, enteritis, pancreatitis, nephritis, rhinitis, atopic dermatitis, heart hypersensitivity, endotoxin shock, central nervous system A pharmaceutical composition comprising an injury or organ (tissue) transplant rejection. A shinto extract having antagonistic activity on platelet activating factor receptor obtained by extracting sinyi with lower alcohol or suspended in distilled water in addition to the above process, washed with hexane and then extracted with ethyl acetate. 8. A shinto extract according to claim 7, comprising a compound of formula 1 and / or a compound of formula 2.

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