JP2011190250A - New compound, antitumor agent and pharmaceutical, food and cosmetic having antitumor action - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a new compound having no risk of side effects, safe, and having excellent antitumor action, to provide antitumor agents having no risk of side effects, safe, and having excellent antitumor action, and to provide pharmaceuticals, foods and cosmetics, having no risk of side effects, safe, and having excellent antitumor action. <P>SOLUTION: There are provided a new compound A represented by formula (1), antitumor agents containing the new compound A, and pharmaceuticals, foods and cosmetics, having antitumor action and containing the new compound A. <P>COPYRIGHT: (C)2011,JPO&INPIT

Description

  The present invention relates to a novel compound, an antitumor agent, and a pharmaceutical, food or cosmetic having an antitumor action.

  Antitumor agents, anticancer agents, and anticancer agents have been selected to act selectively on tumor cells and are relatively less toxic to normal cells. However, leukopenia based on bone marrow suppression, thrombocytopenia, neutrophils The occurrence of fatal side effects such as reduction, side effects of gastrointestinal disorders such as nausea and vomiting, side effects such as hair loss, drug resistance, and restrictions on the route of administration due to many drugs that are difficult to administer orally Many usage problems remain unresolved. Accordingly, there is a demand for antitumor agents, anticancer agents, and anticancer agents that selectively act on tumor cells, have extremely low toxicity to normal cells, and have few restrictions on the administration route.

  Many compounds are known as antitumor agents obtained from natural products. For example, mitomycin C (for example, see Non-patent Document 1), camptothecin (for example, see Patent Document 1), triterpene compound (for example, Patent Document) There are many reports. Further, taxol derivatives (for example, see Patent Document 3), taxane derivatives (for example, see Patent Document 4) and the like are known as antitumor agents using natural products as lead compounds. As for naphthoquinone antitumor agents that are expected to be related to this compound, or related to the biosynthetic system and degradation system of this compound, linacantins A, B, derived from Rhinacanthus nasutus (L.) Kurz There are reports on C, D, G, H, I, K, M, N, and Q (see Non-Patent Document 2, for example). As synthetic compounds related to naphthoquinone antitumor agents, antitumor agents related to furanonaphthoquinone derivatives (for example, see Patent Documents 5 to 8) are known.

  On the other hand, Hakutsukaku Ganoderma (hereinafter referred to as “New Compound A, New Compound B, New Compound C, New Compound D, New Compound E, New Compound F, and New Compound G”), which has been revealed by the present invention Rhinacanthus nasutus (L.) Kurz) is an evergreen small shrub belonging to the genus Linakansaceae, a native of the Deccan Plateau in southern India, and its whole plant is anthelmintic, anti-inflammatory, skin It is known to have an antibacterial action against fungi (see, for example, Non-Patent Document 3), and is mainly used as a Chinese medicine in China, Taiwan, etc., and recently in Japan. In addition, in the previous application by the present applicant, white crane reishi has active oxygen scavenging ability (for example, see Patent Document 9), excretion promoting action (for example, see Patent Document 10), anti-allergy. It is disclosed that there is an action (for example, see Patent Document 11) and that it has an antitumor action (for example, see Patent Document 12).

Japanese Patent No. 348257 Japanese Patent No. 4296312 Japanese Patent Publication No. 6-51689 Japanese Patent No. 3942197 JP-A-9-235280 Japanese Patent Laid-Open No. 9-252753 JP-A-11-21284 JP 2005-220037 A Japanese Patent Laid-Open No. 9-143091 Japanese Patent Laid-Open No. 9-169662 JP 2001-10964 A JP 2002-53481 A

The Journal of Antibiotics, Vol. 9, pages 141-146, 1956 Fight Medicine 49, 2001-2003, 1988 Primary color Makino Wakahan medicinal herb encyclopedia, page 492, Hokuryukan, 1988

  However, the above-mentioned new compound (new compound A, new compound B, new compound C, new compound D, new compound E, new compound F or new compound G), the new compound (new compound A, new compound B, new compound) C, novel compound D, novel compound E, novel compound F or novel compound G) and the novel compound (new compound A, novel compound B, novel compound C, novel compound D, novel compound E, novel compound) There are no known pharmaceuticals, foods or cosmetics containing anti-tumor activity containing compound F or novel compound G).

  Therefore, the present invention provides a novel compound that has no fear of side effects, is safe and has an excellent antitumor effect, an antitumor agent containing the novel compound, and a pharmaceutical agent having the antitumor effect comprising the novel compound The purpose is to provide food or cosmetics.

  As a result of searching for new compounds having antitumor activity, the present inventors have found that seven new compounds (new compound A, new compound B, new compound C, new compound D, new compound) obtained from Hakutsuru Reishi E, novel compound F or novel compound G) was found to have an excellent growth-inhibiting effect on human-derived melanoma cell line (HMV-II), and the present invention was completed. In order to solve the above problems, the present invention comprises the following items.

The present invention relates to “a novel compound represented by the following formula (1) (hereinafter, the novel compound is referred to as a novel compound A”), “an antitumor agent containing the novel compound A”, and “a novel compound A”. Containing a pharmaceutical, food or cosmetic having antitumor activity.

Further, the present invention relates to “a novel compound represented by the following formula (2) (hereinafter, the novel compound is referred to as a novel compound B”), “an antitumor agent containing the novel compound B” and “novel The present invention relates to a pharmaceutical, food or cosmetic containing compound B and having an antitumor action.

Further, the present invention relates to “a novel compound represented by the following formula (3) (hereinafter, the novel compound is referred to as a novel compound C”), “an antitumor agent containing the novel compound C” and “novel The present invention relates to a pharmaceutical, food or cosmetic containing compound C and having an antitumor action.

Further, the present invention relates to “a novel compound represented by the following formula (4) (hereinafter, the novel compound is referred to as a novel compound D”), “an antitumor agent containing the novel compound D” and “novel The present invention relates to a pharmaceutical, food or cosmetic containing compound D and having antitumor activity.

In addition, the present invention relates to “a novel compound represented by the following formula (5) (hereinafter, the novel compound is referred to as a novel compound E”), “an antitumor agent containing the novel compound E”, and “novel The present invention relates to a pharmaceutical, food or cosmetic containing compound E and having an antitumor action.

Further, the present invention relates to “a novel compound represented by the following formula (6) (hereinafter, the novel compound is referred to as a novel compound F”), “an antitumor agent containing the novel compound F” and “novel The present invention relates to a pharmaceutical, food or cosmetic containing compound F and having an antitumor action.

Furthermore, the present invention relates to “a novel compound represented by the following formula (7) (hereinafter, the novel compound is referred to as a novel compound G”), “an antitumor agent containing the novel compound G” and “ The present invention relates to a pharmaceutical, food or cosmetic containing a novel compound G and having an antitumor action.

  According to the present invention, since it is a component derived from a natural product (white crane reishi) that has been conventionally used as a raw material for foods and cosmetics, there is no risk of side effects, and it can be seen from the test examples described later. A novel compound (novel compound A, novel compound B, novel compound C, novel compound D, novel compound E, novel compound F or novel compound G) having an excellent antitumor activity, the novel compound (new compound A, novel compound) B, novel compound C, novel compound D, novel compound E, novel compound F or novel compound G) and the novel compound (novel compound A, novel compound B, novel compound C, novel compound D, novel compound) It is possible to provide a pharmaceutical, food or cosmetic material having an antitumor action, which contains compound E, novel compound F or novel compound G).

It is a flowchart at the time of fractionating / isolating each new compound (new compound A, new compound B, new compound C, new compound D, new compound E, new compound F or new compound G) from Hakutsuru Ganoderma radix. It is a figure which shows the result of having analyzed the three-dimensional configuration of the seven-membered ring of novel compound B by CD (circular dichroism) exciton chirality method.

  Each new compound (new compound A, new compound B, new compound C, new compound D, new compound E, new compound F or new compound G) of the present invention is obtained by extraction / purification process using Hakutsuru Reishi as a raw material. It may also be obtained from other plants. In addition, a new compound (new compound A, new compound B, new compound C, new compound D, new compound E, new compound F or new compound G) obtained by synthesis can also be used. In addition, extracts from white crane reishi grass and other plants containing a new compound (new compound A, new compound B, new compound C, new compound D, new compound E, new compound F or new compound G), A crude product, a dried plant body, or a plant paste can also be used.

  When extracting and purifying the novel compound (new compound A, new compound B, new compound C, new compound D, new compound E, new compound F or new compound G) of the present invention from plants such as white crane reishi grass Any extraction / purification process used industrially can be used. The plant leaves, stems, roots, flowers, etc., which are raw materials, are collected at an appropriate time, and then directly or subjected to a drying process such as ventilation drying to obtain extraction raw materials. When extracting from said dried plant body, it can carry out with reference to a well-known method (Phytomedicine 16th volume, 929-934 pages 2009).

  That is, the raw material is pulverized or chopped and then extracted using a solvent. Extraction solvents include water, alcohols such as ethanol, methanol, isopropyl alcohol, ketones such as acetone and methyl ethyl ketone, esters such as methyl acetate and ethyl acetate, and lipophilic solvents such as hexane and chloroform, either alone or in combination. It can be used as a solvent. Extraction temperature is 0-100 degreeC normally, Preferably it is 5-50 degreeC. The extraction time is about 1 hour to 10 days, and the amount of solvent is usually 1 to 30 times weight, preferably 5 to 10 times weight per dry raw material. The extraction operation may be performed by stirring or by immersion. The extraction operation may be repeated 2-3 times as necessary. Each novel compound (new compound A, new compound B, new compound C, new compound D) from the extract obtained by removing insoluble residues from the crude extract obtained by the above operation by filtration or centrifugation, or from the juice of a plant The novel compound E, the novel compound F or the novel compound G) may be purified by any known crude drug separation / purification method, but may be any of two-phase solvent distribution method, countercurrent distribution method, column chromatography. It is preferable to use a method, a preparative high performance liquid chromatography method or the like alone or in combination. For example, as a two-phase solvent partitioning method, a method of recovering a target compound to a solvent phase by partitioning a solvent such as n-hexane, chloroform, methyl ethyl ketone, ethyl acetate, or methyl acetate with water from the above extract can be mentioned. . Examples of column chromatography include ion exchange column chromatography, normal phase or reverse phase silica gel as a carrier, adsorption column chromatography using Diaion HP-20, etc., and Sephadex LH-20 as a carrier. Gel filtration methods using modified dextran gels such as these can be mentioned, and these can be used alone or in combination or repeatedly. Examples of the preparative high performance liquid chromatography method include a method using a reverse phase column using octadecyl silica and the like, a method using a normal phase column using silica gel and the like.

  The administration route of the antitumor agent of the present invention is not particularly limited, and examples thereof include enteral administration such as oral administration and rectal administration, mucosal administration such as nasal administration, injection administration such as intravenous administration and subcutaneous administration, etc. Can give. The dosage form of the antitumor agent of the present invention can be in the form of a preparation suitable for the administration method, for example, tablet, powder, fine granule, granule, capsule, powder, pill, troche And liquids such as solid preparations, solutions, suspensions, emulsions, syrups, injections, and gel preparations. Pure products, purified products, crude products, etc. of each new compound (new compound A, new compound B, new compound C, new compound D, new compound E, new compound F or new compound G) may be administered as they are. However, it may be administered together with a pharmacologically acceptable excipient. As the excipient, any monosaccharides, second class, polysaccharides, inorganic salts, fats and oils, distilled water, etc. that can be generally used as preparations can be used. In formulating, additives such as binders, lubricants, dispersants, suspending agents, emulsifiers, diluents, buffers, antioxidants, and bacterial inhibitors can be used.

  The effective dose of each new compound (new compound A, new compound B, new compound C, new compound D, new compound E, new compound F or new compound G) is the administration route, dosage form, disease symptom, subject Usually, it is 0.1 to 1000 mg, preferably 0.5 to 300 mg, more preferably 1 to 100 mg per day per adult. The content of each novel compound (novel compound A, novel compound B, novel compound C, novel compound D, novel compound E, novel compound F or novel compound G) in the oral antitumor agent of the present invention is the form of the preparation -Based on the data of the effective dose and the dose as the preparation, the optimal active ingredient content in the preparation can be set for each dosage form.

  As a form of food, a dried plant body containing white crane reishi grass and each new compound (new compound A, new compound B, new compound C, new compound D, new compound E, new compound F or new compound G) As a tea, a pure product of each new compound (new compound A, new compound B, new compound C, new compound D, new compound E, new compound F or new compound G), or a partially purified product of the new compound And a crude extract of the novel compound from the plant containing the novel compound, a plant paste containing the novel compound, and a food containing a dried plant product containing the novel compound.

  As tea, you may use it individually or in mixture with another tea raw material. Other tea ingredients include green tea, oolong tea, puer tea, black tea, roasted green tea, brown rice tea, Tochu tea, kashiwanoha tea, mulberry leaf tea, etc. Can also be used.

  When obtaining a plant extract, any method can be used as long as it is a method usually used for extraction of food such as extraction with hot water, extraction with ethanol or hydrous ethanol. Crude extracts and partially purified products from plants of each new compound (new compound A, new compound B, new compound C, new compound D, new compound E, new compound F or new compound G) should be obtained by conventional methods. Can do.

  As the form of the food having antitumor activity of the present invention, in addition to tea, drinks, jelly, biscuits, tablets, pills, soft capsules, hard capsules, powders, fine granules, granules, etc. are provided as normal foods Any form can be used as long as it is possible. Additives such as excipients, binders, lubricants, dispersants, suspending agents, emulsifiers, diluents, buffers, antioxidants, and bacterial inhibitors can also be used as auxiliary materials.

  The effective intake amount of each new compound (new compound A, new compound B, new compound C, new compound D, new compound E, new compound F or new compound G) by the food having antitumor action of the present invention is the ingestion form Usually, it is 0.001 to 100 mg, preferably 0.01 to 10 mg, more preferably 0.1 to 1 mg per day for an adult, although it varies depending on the health condition of the subject, the age of the subject, and the like.

  The content of each new compound (new compound A, new compound B, new compound C, new compound D, new compound E, new compound F or new compound G) in the food having antitumor activity of the present invention is Although it varies depending on the form, it is usually 0.0001 to 1 wt%, preferably 0.001 to 0.5 wt%, more preferably 0.01 to 0.1 wt%.

  It does not specifically limit as an example of the form of the external medicine and cosmetics of this invention.

  Examples of the form of external medicine include ointments, creams, cloths, tapes, and external preparations. The pharmaceutical product of the present invention contains other pharmaceutical components as necessary in each new compound (new compound A, new compound B, new compound C, new compound D, new compound E, new compound F or new compound G). In addition, additives such as a binder, a dispersant, a suspending agent, an emulsifier, a diluent, a buffer, an antioxidant, and a bacteria inhibitor may be used.

  As the form of the cosmetic, any form that can be used as an external preparation / cosmetic preparation such as lotion, cosmetic liquid, milky lotion, cream, gel, pack, cosmetic powder, facial cleansing foam, bath preparation, etc. can be used. The cosmetics include a pure product of each new compound (new compound A, new compound B, new compound C, new compound D, new compound E, new compound F or new compound G), a partially purified product of the new compound, In addition to essential components such as a crude extract of the novel compound from the plant or a plant containing the novel compound, it may contain components to be blended in the cosmetic preparation as necessary. Examples of the ingredients include solid oil, semi-solid oil, liquid oil, low molecular moisturizer, polymer moisturizer, fat-soluble moisturizer, emollient, surfactant, preservative, antioxidant, pH adjuster, Ethanol, water, etc. can be raised.

  The effective dosage of each new compound (new compound A, new compound B, new compound C, new compound D, new compound E, new compound F or new compound G) for external use is the subject's symptoms and the subject's age. Usually, it is 0.001 to 100 mg, preferably 0.01 to 10 mg, more preferably 0.1 to 1 mg per day for an adult, although it varies depending on the like.

  The content of each new compound (new compound A, new compound B, new compound C, new compound D, new compound E, new compound F or new compound G) in the antitumor agent, pharmaceuticals and cosmetics of the present invention is: It is 0.0001-1 wt% normally or individually as a mixture, preferably 0.001-0.5 wt%, more preferably 0.01-0.1 wt%.

  Hereinafter, the present invention will be described in more detail by giving examples of fractionation and isolation of each novel compound of the present invention and linacantin C as a comparative example, test examples and examples of antitumor activity. It is not restricted.

1. Fractionation and isolation of each novel compound of the present invention and linacantin C as a comparative example 1-1. New compound A
(1) Fractionation / isolation of novel compound A Fractionation / isolation of novel compound A was performed according to the flow shown in FIG. That is, dry roots (Rhinacanthus nasutus (L.) Kurz) (5 Kg) were extracted with 25 L of 90% (v / v) ethanol three times at room temperature for 24 hours each. They were combined and concentrated to give a dried product (407 g). Next, this was suspended and dissolved in 7 L of 90% (v / v) methanol, then distributed three times with an equal amount of hexane, and then the 90% (v / v) methanol phase was taken and concentrated under reduced pressure. Purified water was added to the concentrate under reduced pressure, filled up to 5 L, transferred to a separatory funnel, and two-phase solvent partition was performed with chloroform three times. Next, the chloroform phases obtained by this operation were combined to obtain 69.3 g of a dried product.

  Of this, 69.0 g was subjected to silica gel column chromatography (80 mmφ × 150 mm, manufactured by Kanto Chemical Co., Inc.) using hexane / ethyl acetate as an elution solvent. 3 bed volume (BV) elution solvent hexane / ethyl acetate (9: 1), 2 BV elution solvent hexane / ethyl acetate (8: 2), and 2 BV elution solvent hexane / ethyl acetate (7: 3 The silica gel column was washed successively with 2) and then eluted with 2 BV elution solvent hexane / ethyl acetate (6: 4) to obtain fraction C (dry solid: 3.73 g).

Next, Sephadex LH-20 column chromatography (20 mmφ × 200 mm, manufactured by Pharmacia) using methanol as an elution solvent was performed on fraction C. That is, the column was washed with 2 BV methanol and then eluted with 1 BV methanol to obtain fraction F (dry solid 369 mg).
Further, fraction F (dry solid 369 mg) was subjected to flash ODS column chromatography (20 mmφ × 150 mm, Nomura Chemical Co., Ltd.) using water / methanol as an elution solvent. That is, after washing the flash ODS column with 180 ml of 50% (v / v) methanol, stepwise wash sequentially with 60% (v / v) methanol, 70% (v / v) methanol, then 80 Elution with% (v / v) methanol gave fraction G (dry solid 75.6 mg) containing the desired novel compound A.
Further, fraction G (dried solid 75.6 mg) was subjected to preparative high performance liquid chromatography (ODS column, 20 mmφ × 250 mm, manufactured by Nomura Chemical Co., Ltd., mobile phase: 45% (v / v) acetonitrile, detection: 254 nm UV The product was purified with a monitor) to obtain a novel compound A (dried product 9.8 mg).

  In the above, Waters 515 system and Waters 600 system (both manufactured by Nippon Waters Co., Ltd.) were used for the high performance liquid chromatography. For silica gel column chromatography, Sephadex LH-20 column chromatography and flash ODS chromatography, general-purpose laboratory instruments and experimental devices were used.

(2) Structural analysis of novel compound A The structural analysis of novel compound A was performed using high-resolution mass spectrometry (HRFABMS) and nuclear magnetic resonance spectroscopy ( ¹ H NMR, ¹³ C NMR). The results are shown below.

(2-1) Results by high-resolution mass spectrometry (HRFABMS) In high-resolution mass spectrometry (HRFABMS), “m / z 272.1039 [M] ⁺ (calcd. 272.1049 Δ 0.9 mmu)” was observed, and the molecular formula Was found to be “C ₁₆ H ₁₆ O ₄ ”. In the low resolution mass spectrometry (LRFABMS), “m / z 272.” was observed.

(2-2) In the nuclear magnetic resonance spectroscopy ⁽¹ H NMR) Results from nuclear magnetic resonance spectroscopy ⁽¹ H NMR), the following peaks were observed.
^“1 H NMR (CDCl _3, 500 MHz): δ 3.69 (H-14, J = 11.2 Hz, 1H, d), 3.75 (H-14, J = 11.2 Hz, 1H, d), 3.95 (8-OMe , 3H, s), 4.22 (H-2, J = 11.7 Hz, 1H, d), 4.31 (H-2, J = 11.7 Hz, 1H, d), 5.93 (H-4, J = 11.7 Hz, 1H , d), 6.43 (H-5, J = 11.7 Hz, 1H, d), 6.51 (H-7, 1H, s), 7.47 (H-10, J = 1.5, 6.7, 7.5 Hz, 1H, m) , 7.50 (H-11, J = 1.5, 6.7, 7.5 Hz, 1H, m), 8.14 (H-9, J = 1.5, 7.5 Hz, 1H, d), 8.24 (H-12, J = 1.5, 7.5 Hz, 1H, d). ''

(2-3) In the nuclear magnetic resonance spectroscopy ⁽¹³ C NMR) Results from nuclear magnetic resonance spectroscopy ⁽¹³ C NMR), the following peaks were observed.
^“13 C NMR (CDCl _3, 125 MHz): δ 55.7 (8-OMe), 65.9 (C-14), 74.4 (C-2), 74.9 (C-3), 106.8 (C-7), 119.1 ( C-6), 121.6 (C-9), 122.7 (C-12), 126.0 (C-12a), 126.3 (C-10), 126.7 (C-11), 127.6 (C-8a), 130.1 (C -5), 132.1 (C-4), 148.9 (C-13), 150.6 (C-8). ''

In the above, a JEOL JMS SX-102 type mass spectrometer (manufactured by JEOL Ltd.) was used as the high resolution mass spectrometer. Moreover, as a nuclear magnetic resonance spectrometer ( ¹ H NMR and ¹³ C NMR), a JEOL JNM-GSX500 type nuclear magnetic resonance spectrometer (manufactured by JEOL Ltd.) was used.

  From the above results and the HMQC spectrum and HMBC spectrum, it was found that the novel compound A was a novel compound represented by the above formula (1).

1-2. New compounds B and C
(1) Fractionation / isolation of novel compounds B and C Fractionation / isolation of novel compounds B and C were carried out according to the flow shown in FIG. That is, dry roots (Rhinacanthus nasutus (L.) Kurz) (5 Kg) were extracted with 25 L of 90% (v / v) ethanol three times at room temperature for 24 hours each. They were combined and concentrated to give a dried product (407 g). Next, this was suspended and dissolved in 7 L of 90% (v / v) methanol, then distributed three times with an equal amount of hexane, and then the 90% (v / v) methanol phase was taken and concentrated under reduced pressure. Purified water was added to the concentrate under reduced pressure, filled up to 5 L, transferred to a separatory funnel, and two-phase solvent partition was performed with chloroform three times. Next, the chloroform phases obtained by this operation were combined to obtain 69.3 g of a dried product.

  Of this, 69.0 g was subjected to silica gel column chromatography (80 mmφ × 150 mm, manufactured by Kanto Chemical Co., Inc.) using hexane / ethyl acetate as an elution solvent. That is, the silica gel column was washed with 3 BV elution solvent hexane / ethyl acetate (9: 1) and then eluted with 1 BV elution solvent hexane / ethyl acetate (8: 2). Got.

Next, the fraction A was subjected to Sephadex LH-20 column chromatography (20 mmφ × 200 mm, manufactured by Pharmacia) using methanol as an elution solvent. That is, the Sephadex LH-20 column was washed with 1.5 BV methanol and then eluted with 0.5 BV methanol to obtain a fraction A-2 (1.34 g).
Further, fraction A-2 was fractionated by flash ODS column chromatography (20 mm φ × 150 mm, manufactured by Wako Pure Chemical Industries, Ltd.) using water / methanol as an elution solvent. That is, after washing the flash ODS column with 180 ml of 50% (v / v) methanol, stepwise wash sequentially with 60% (v / v) methanol, 70% (v / v) methanol, then 80 Elution with% (v / v) methanol gave fraction A-2-1 (261 mg of dry solid) containing the desired novel compounds B and C.
Fraction A-2-1 was further fractionated by silica gel column chromatography (20 mmφ × 150 mm, manufactured by Kanto Chemical Co., Inc.) using hexane / ethyl acetate as an elution solvent. Specifically, after washing the silica gel column with 3 BV hexane / ethyl acetate (9: 1) elution solvent, elution was performed with 1 BV hexane / ethyl acetate (8: 2) elution solvent, and fraction D (dried solid 92.4 mg )

  Further, fraction D (dried product 92.4 mg) was subjected to preparative high performance liquid chromatography (ODS column, 20 mmφ × 250 mm, Nomura Chemical Co., mobile phase: 58% (v / v) acetonitrile / water / 0.1%. Purification by (v / v) formic acid, detection: 254 nm UV monitor) gave new compound B (4.6 mg dried product) and new compound C (8.0 mg dried product).

(2) Structural analysis of new compounds B and C The structural analysis of new compounds B and C is performed using high resolution mass spectrometry (HRFABMS) and nuclear magnetic resonance spectroscopy ( ¹ H-NMR, ¹³ C-NNMR). It was. The results are shown below.

(2-1) Result by high resolution mass spectrometry (HRFABMS) (2-1-1) Novel compound B
With respect to the novel compound B, “m / z 256.1099 [M] ⁺ (calcd. 256.1010 Δ0 mmu).” Was observed, and it was found that the molecular formula was “C ₁₆ H ₁₆ O ₃ ”. In LRFABMS, "m / z 256." was observed, and in LREIMS, "m / z 256." was observed.
(2-1-2) Novel compound C
With respect to the novel compound C, “m / z 270.1266 [M] ⁺ (calcd. 270.1256 Δ1.0 mmu)” was observed, and it was found that the molecular formula was “C ₁₇ H ₁₈ O ₃ ”. In LRFABMS, “m / z 270” was observed.

(2-2) Results by nuclear magnetic resonance spectroscopy ( ¹ H NMR) (2-2-1) New compound B
For the new compound B, the following peaks were observed.
^“1 H NMR (CDCl _3, 500 MHz): δ 1.31 (H-14, 3H, s), 3.89 (H-2, J = 11.7 Hz, 1H, d), 3.90 (8-OMe, 3H, s) 4.29 (H-2, J = 2.0, 11.7 Hz, 1H, d), 5.97 (H-4, J = 2.0, 11.7 Hz, 1H, dd), 6.26 (H-5, J = 11.7 Hz, 1H, d), 6.48 (H-7, 1H, s), 7.42 (H-10, J = 6.3, 7.8 Hz, 1H, m), 7.46 (H-11, J = 6.3, 7.8 Hz, 1H, m), 8.10 (H-9, J = 7.8 Hz, 1H, d), 8.20 (H-12, J = 7.8 Hz, 1H, d). ''
(2-2-2) Novel compound C
For the new compound C, the following peaks were observed.
^“1 H NMR (CDCl _3, 500 MHz): δ 1.32 (H-14, 3H, s), 3.38 (3-OMe, 3H, s), 3.91 (8-OMe, 3H, s), 4.08 (H- 2, J = 11.7 Hz, 1H, d), 4.33 (H-2, J = 11.7 Hz, 1H, d), 5.90 (H-4, J = 11.7 Hz, 1H, d), 6.42 (H-5, J = 11.7 Hz, 1H, d), 6.50 (H-7, 1H, s), 7.40 (H-10, 1H, m), 7.44 (H-11, 1H, m), 8.08 (H-9, J = 7.8 Hz, 1H, d), 8.20 (H-12, J = 7.8 Hz, 1H, d). ''

(2-3) Results obtained by nuclear magnetic resonance spectroscopy ( ¹³ C NMR) (2-3-1) Novel compound B
For the new compound B, the following peaks were observed.
^“13 C NMR (CDCl _3, 125 MHz): δ 23.9 (C-14), 55.7 (8-OMe), 72.9 (C-3), 79.5 (C-2), 106.55 (C-7), 119.9 ( C-6), 121.7 (C-9), 122.6 (C-12), 125.8 (C-12a), 126.2 (C-10), 126.6 (C-11), 127.2 (C-5), 127.6 (C -8a), 136.8 (C-4), 149.2 (C-13), 150.8 (C-8). ''
(2-3-2) Novel compound C
For the new compound C, the following peaks were observed.
^“13 C NMR (CDCl _3, 125 MHz): δ 24.3 (C-14), 52.1 (3-OMe), 55.7 (8-OMe), 77.3 (C-3), 77.5 (C-2), 106.9 ( C-7), 119.5 (C-6), 121.6 (C-9), 122.8 (C-12), 126.1 (C-12a), 126.5 (C-10), 127.2 (C-11), 127.7 (C -8a), 129.2 (C-5), 134.8 (C-4), 148.9 (C-13), 150.5 (C-8). ''

(2-4) Determination of configuration of seven-membered 3-position asymmetric carbon of new compound B:
To a tetrahydrofuran solution (500 μL) of novel compound B (5.0 mg, 20 μmol) was added a hexane solution (1.6 M, 20 μL) of n-butyllithium at −78 ° C. in a nitrogen atmosphere. After stirring for 15 minutes, benzoyl chloride (10 mg, 71 μmol) was added to the reaction solution and stirred at −78 ° C. in a nitrogen atmosphere. After 15 minutes, the reaction solution was warmed to room temperature and further stirred for 30 minutes. Next, a saturated aqueous sodium hydrogen carbonate solution was added to the reaction solution, followed by extraction three times with an equal amount of ethyl acetate, and the ethyl acetate phase was combined. Magnesium sulfate was added to the resulting ethyl acetate phase for dehydration. Further, the ethyl acetate solvent was distilled off under reduced pressure, and the resulting dried product was subjected to silica gel column chromatography (10 mmΦ x 80 mm, manufactured by Kanto Chemical Co., Inc.) using hexane / ethyl acetate as an elution solvent for fractionation. did. In other words, the ethyl acetate phase dried product obtained above was applied to a silica gel column previously equilibrated with hexane / ethyl acetate (9: 1), and the silica gel column was added with 10 mL of hexane / ethyl acetate (9: 1). After elution with 3 BV (= 18.8 mL) of elution solvent hexane / ethyl acetate (9: 1), benzoylated new compound B (2.6 mg) was obtained. The new benzoylated compound B was confirmed by ¹ H NMR spectrum, ¹³ C NMR spectrum, and two-dimensional NMR spectrum (COSY, HMQC, HMBC).

The configuration of the 3-position of the seven-membered ring of the novel benzoylated compound B was analyzed by CD (circular dichroism) exciton chirality method. That is, as a CD spectrometer, a JASCO J-820 type circular dichroism dispersometer (manufactured by JASCO Corporation) was used, and the benzoylated novel compound B was dissolved in methanol and measured, and analyzed by measuring the Cotton effect.
FIG. 2 is a diagram showing the result of analyzing the configuration of the 3-position of the seven-membered ring of the novel compound B by the CD (circular dichroism) exciton chirality method. As can be seen from FIG. 2, in the CD spectrum, a negative cotton effect was observed at 266 nm and a positive cotton effect at 230 nm. Therefore, the exciton chirality of the new benzoylated Compound B was determined to be counterclockwise (negative chirality), and the configuration of the asymmetric carbon at the 7-membered ring 3-position was determined to be S.

  From the above results and the HMQC spectrum and HMBC spectrum, the new compound B is a new compound represented by the above formula (2), and the new compound C is a new compound represented by the above formula (3). I understood that.

From the determination of the configuration of the seven-membered 3-position asymmetric carbon of the new compound B described in (2-4) above, the new compound B is a new compound represented by the following formula (2a) I understand.

In the above (2-4), the benzoylation reaction of the novel compound B is according to the following reaction formula.

1-3. New compounds D and E
(1) Fractionation and isolation of novel compounds D and E Fractionation and isolation of novel compounds D and E were carried out according to the flow shown in FIG. That is, dry roots (Rhinacanthus nasutus (L.) Kurz) (5 Kg) were extracted with 25 L 90% (v / v) ethanol three times at room temperature for 24 hours each. They were combined and concentrated to give a dried product (407 g). Next, this was suspended and dissolved in 7 L of 90% (v / v) methanol, then distributed three times with an equal amount of hexane, and then the 90% (v / v) methanol phase was taken and concentrated under reduced pressure. Purified water was added to the concentrate under reduced pressure, filled up to 5 L, transferred to a separatory funnel, and two-phase solvent partition was performed with chloroform three times. Next, the chloroform phases obtained by this operation were combined to obtain 69.3 g of a dried product.

  Of this, 69.0 g was subjected to silica gel column chromatography (80 mmφ × 150 mm, manufactured by Kanto Chemical Co., Inc.) using hexane / ethyl acetate as an elution solvent. Specifically, the column was sequentially washed with 3 BV elution solvent hexane / ethyl acetate (9: 1) and 2 BV elution solvent hexane / ethyl acetate (8: 2), and then 2 BV elution solvent hexane / ethyl acetate (7 : Elution by 3) gave fraction B (dried product: 5.66 g).

Next, the fraction B was subjected to Sephadex LH-20 column chromatography (20 mmφ × 200 mm, manufactured by Pharmacia) using methanol as an elution solvent. That is, the column was washed with 2.5 BV methanol and then eluted with 1 BV methanol to obtain fraction B-2 (dried product 480 mg).
Further, fraction B-2 (dry solid 480 mg) was subjected to flash ODS column chromatography (20 mmφ × 150 mm, manufactured by Nomura Chemical Co., Ltd.) using water / methanol as an elution solvent. That is, after washing the flash ODS column with 180 ml of 50% (v / v) methanol, stepwise, 60% (v / v) methanol, 70% (v / v) methanol, 80% (v / v) v) After washing with methanol and elution with 90% (v / v) methanol, fraction X (dry solid 143 mg) containing the target novel compounds D and E was obtained.

Further, fraction X (dry solid 143 mg) was fractionated by silica gel column chromatography (20 mmφ × 150 mm, manufactured by Kanto Chemical Co., Inc.). Specifically, after washing the silica gel column with 2 BV elution solvent hexane / ethyl acetate (8: 2), elution with 1 BV elution solvent hexane / ethyl acetate (7: 3), fraction B-2-1 ( Dry solid (32.3 mg) and fraction B-2-2 (dry solid 18.8 mg) were obtained.
From fraction B-2-1 (dried solid 32.3 mg), preparative high performance liquid chromatography (ODS column, 20 mmφ × 250 mm, Nomura Chemical Co., mobile phase: 58% (v / v) acetonitrile, Detection: 280 nm UV monitor) to obtain a novel compound D (dry solid 5.0 mg).
From fraction B-2-2 (dried solid 18.8 mg), preparative high performance liquid chromatography (ODS column, 20 mmφ × 250 mm, Nomura Chemical Co., mobile phase: 70% (v / v) methanol, Detection: 280 nm UV monitor), preparative high performance liquid chromatography (ODS column, 20 mmφ × 250 mm, Nomura Chemical, mobile phase: 55% (v / v) acetonitrile, detection: 254 nm UV monitor) Purification was performed to obtain a new compound E (dry solid 1.6 mg).

(2) Structural analysis of the new compounds D and E The structural analysis of the new compounds D and E was performed using high resolution mass spectrometry (HRFABMS) and nuclear magnetic resonance spectroscopy ( ¹ H NMR, ¹³ C NMR). The results are shown below.

(2-1) Result by high resolution mass spectrometry (HRFABMS) (2-1-1) Novel compound D
With respect to the novel compound D, “m / z 397.1651 [M + H] ⁺ (calcd. 397.1651 Δ 0.0 mmu)” was observed, and it was found that the molecular formula was “C ₂₃ H ₂₄ O ₆ ”.
(2-1-2) Novel compound E
With respect to the novel compound E, “m / z 441.1919 [M + H] ⁺ (calcd. 441.1913 Δ0.5 mmu).” Was observed, and it was found that the molecular formula was “C ₂₅ H ₂₈ O ₇ ”.

(2-2) Results by nuclear magnetic resonance spectroscopy ( ¹ H NMR) (2-2-1) New compound D
In the new compound D, the following peaks were observed.
^"1 H NMR (CDCl _3, 500 MHz): δ 0.98 (H-12 and H-13, 6H, s), 1.98 (H-8 ', 3H, s), 2.24 (H-7', 3H, s) ), 2.65 (H-9, 2H, s), 3.90 (H-11, 2H, s), 6.21 (H-5 ', 1H, d), 7.08 (H-3', 1H, d), 7.25 ( H-4 ', 1H, dd), 7.60 (H-7, 1H, t), 7.67 (H-6, 1H, t), 7.99 (H-8, 1H, d), 8.03 (H-5, 1H , d).
(2-2-2) Novel compound E
In the new compound E, the following peaks were observed.
^“1 H NMR (CDCl _3, 500 MHz): δ 1.01 (H-12 and H-13, 6H, s), 1.48 (H-10 ′, 3H, s), 1.90 (H-9 ′, 3H, s) ), 2.26 (H-8 ', 3H, s), 2.70 (H-9, 2H, s), 3.91 (H-11, 2H, s), 6.02 (H-5', 1H, d), 6.67 ( H-4 ', 1H, dd), 7.09 (H-3', 1H, d), 7.66 (H-7, 1H, t), 7.73 (H-6, 1H, t), 8.04 (H-8, 1H, d), 8.08 (H-5, 1H, d). ''

(2-3) Results by nuclear magnetic resonance spectroscopy (13C-NMR) (2-3-1) Novel compound D
In the new compound D, the following peaks were observed.
^“13 C NMR (CDCl _3, 125 MHz): δ 13.4 (C-8 ′), 25.3 (C-12 and C-13), 28.1 (C-7 ′), 32.2 (C-9), 37.0 (C -10), 73.4 (C-11), 121.6 (C-3), 126.1 (C-8), 127.0 (C-5), 129.3 (C-8a), 132.9 (C-4a), 132.9 (C- 7), 134.8 (C-5 '), 134.9 (C-3'), 135.0 (C-6), 136.2 (C-2 '), 136.4 (C-4'), 154.4 (C-2), 167.3 (C-1 '), 181.3 (C-1), 184.8 (C-4), 198.0 (C-6'). ''
(2-3-2) Novel compound E
In the new compound E, the following peaks were observed.
^“13 C NMR (CDCl _3, 125 MHz): δ 12.8 (C-9 ′), 22.6 (C-8 ′), 25.2 (C-12 and C-13), 25.3 (C-10 ′), 32.1 ( C-9), 37.2 (C-10), 73.0 (C-11), 79.4 (C-6 '), 121.4 (C-3), 126.1 (C-8), 126.4 (C-4'), 127.0 (C-5), 129.1 (C-8a), 132.9 (C-7), 133.3 (C-4a), 134.9 (C-6), 136.3 (C-3 '), 136.5 (C-2'), 139.8 (C-5 '), 153.4 (C-2), 167.7 (C-1'), 181.6 (C-1), 184.2 (C-4), 207.3 (C-7 ').''

  From the above results and HMQC spectrum and HMBC spectrum, the new compound D is a new compound represented by the above formula (4), and the new compound E is a new compound represented by the above formula (5). I understood that.

1-4. New compound F
(1) Fractionation / isolation of novel compound F Fractionation / isolation of novel compound F was performed according to the flow shown in FIG. That is, dry roots (Rhinacanthus nasutus (L.) Kurz) (5 Kg) were extracted with 25 L 90% (v / v) ethanol three times at room temperature for 24 hours each. They were combined and concentrated to give a dried product (407 g). Next, this was suspended and dissolved in 7 L of 90% (v / v) methanol, then distributed three times with an equal amount of hexane, and then the 90% (v / v) methanol phase was taken and concentrated under reduced pressure. Purified water was added to the concentrate under reduced pressure, filled up to 5 L, transferred to a separatory funnel, and two-phase solvent partition was performed with chloroform three times. Next, the chloroform phases obtained by this operation were combined to obtain 69.3 g of a dried product.

  Of this, 69.0 g was subjected to silica gel column chromatography (80 mmφ × 150 mm, manufactured by Kanto Chemical Co., Inc.) using hexane / ethyl acetate as an elution solvent. Specifically, the column was sequentially washed with 3 BV elution solvent hexane / ethyl acetate (9: 1) and 2 BV elution solvent hexane / ethyl acetate (8: 2), and then 2 BV elution solvent hexane / ethyl acetate (7 : Elution by 3) gave fraction B (dried product: 5.66 g).

Next, Sephadex LH-20 column chromatography (20 mmφ × 200 mm, manufactured by Pharmacia) was performed on fraction B (dried product 2.8 g) using methanol as an elution solvent. That is, the column was washed with 1.5 BV methanol and eluted with 0.5 BV methanol to obtain fraction B-1 (dried solid 1.16 g).
Further, fraction B-1 (1.1 g of dried product) was subjected to flash ODS column chromatography (20 mmφ × 100 mm, manufactured by Nomura Chemical Co., Ltd.) using water / methanol as an elution solvent. That is, after washing the flash ODS column with 180 ml of 50% (v / v) methanol, stepwise wash sequentially with 60% (v / v) methanol, 70% (v / v) methanol, then 80 Elution with% (v / v) methanol gave fraction B-1-2 (dry solid 429 mg) containing the desired novel compound F.
Next, fraction B-1-2 (dried product 429 mg) was fractionated by silica gel column chromatography (20 mmφ × 150 mm, manufactured by Kanto Chemical Co., Inc.) using hexane / ethyl acetate as an elution solvent. Specifically, after washing the silica gel column with 2 BV elution solvent hexane / ethyl acetate (8: 2), elution with 1 BV elution solvent hexane / ethyl acetate (7: 3), fraction B-1-2 2 (dried product 32.9 mg) was obtained.
Fraction B-1-2-2 (dried product 32.9 mg) was subjected to preparative high performance liquid chromatography (ODS column, 20 mmφ × 250 mm, manufactured by Nomura Chemical Co., Ltd., mobile phase: 55% (v / v) acetonitrile, Detection: 280 nm UV monitor), preparative high performance liquid chromatography (C-30 column, 20 mmφ × 250 mm, Nomura Chemical, mobile phase: 55% (v / v) acetonitrile, detection: 254 nm UV monitor) to obtain a novel compound F (dried solid 10.3 mg).

(2) Structural analysis of the novel compound F The structural analysis of the novel compound F was performed using high resolution mass spectrometry (HRFABMS) and nuclear magnetic resonance spectroscopy ( ¹ H NMR, ¹³ C NMR). The results are shown below.

(2-1) Result by high resolution mass spectrometry (HRFABMS) In high resolution mass spectrometry (HRFABMS), “m / z 459.2379 [M + H] ⁺ (calcd. 459.2383 Δ 0.4 mmu)” was observed. The molecular formula was found to be “C ₂₆ H ₃₄ O ₇ ”.

(2-2) In the nuclear magnetic resonance spectroscopy ⁽¹ H NMR) Results from nuclear magnetic resonance spectroscopy ⁽¹ H NMR), the following peaks were observed.
^"1 H NMR (CDCl _3, 500 MHz): δ 1.00 (H-12 and H-13, 6H, s), 1.08 (H-10 ', 3H, s), 1.13 (H-8', 3H, d ), 1.46 (H-5 ', 1H, m), 1.66 (H-5', 1H, m), 1.79 (H-9 ', 3H, s), 2.08 (H-4', 1H, m), 2.23 (H-4 ', 1H, m), 2.69 (H-9, 2H, s), 3.20 (6'-OMe, 3H, s), 3.80 (H-7', 1H, q), 3.89 (H -11, 2H, s), 6.70 (H-3 ', 1H, t), 7.66 (H-7, 1H, t), 7.73 (H-6, 1H, t), 8.06 (H-8, 1H, d), 8.09 (H-5, 1H, d). ''

(2-3) In the nuclear magnetic resonance spectroscopy ⁽¹³ C NMR) Results from nuclear magnetic resonance spectroscopy ⁽¹³ C NMR), the following peaks were observed.
^“13 C NMR (CDCl _3, 125 MHz): δ 12.3 (C-9 ′), 17.0 (C-8 ′), 18.6 (C-10 ′), 22.7 (C-4 ′), 25.2 (C-12 ), 25.2 (C-13), 31.9 (C-5 '), 32.2 (C-9), 37.0 (C-10), 49.1 (6'-OMe), 70.6 (C-7'), 72.8 (C -11), 78.4 (C-6 '), 121.8 (C-3), 126.1 (C-8), 127.0 (C-5), 127.7 (C-2'), 129.4 (C-8a), 132.9 ( C-7), 133.0 (C-4a), 134.9 (C-6), 142.4 (C-3 '), 154.2 (C-2), 168.1 (C-1'), 181.2 (C-1), 184.8 (C-4). ''

  From the above results and the HMQC spectrum and HMBC spectrum, it was found that the new compound F was a new compound represented by the above formula (6).

1-5. New compound G
(1) Fractionation / isolation of novel compound G Fractionation / isolation of novel compound G was performed according to the flow shown in FIG. That is, dry roots (Rhinacanthus nasutus (L.) Kurz) (5 Kg) were extracted with 25 L 90% (v / v) ethanol three times at room temperature for 24 hours each. They were combined and concentrated to give a dried product (407 g). Next, this was suspended and dissolved in 7 L of 90% (v / v) methanol, then distributed three times with an equal amount of hexane, and then the 90% (v / v) methanol phase was taken and concentrated under reduced pressure. Purified water was added to the concentrate under reduced pressure, filled up to 5 L, transferred to a separatory funnel, and two-phase solvent partition was performed with chloroform three times. Next, the chloroform phases obtained by this operation were combined to obtain 69.3 g of a dried product.

  Of this, 69.0 g was subjected to silica gel column chromatography (80 mmφ × 150 mm, manufactured by Kanto Chemical Co., Inc.) using hexane / ethyl acetate as an elution solvent. Specifically, the column was sequentially washed with 3 BV elution solvent hexane / ethyl acetate (9: 1) and 2 BV elution solvent hexane / ethyl acetate (8: 2), and then 2 BV elution solvent hexane / ethyl acetate (7 : Elution by 3) gave fraction B (dried product: 5.66 g).

Next, Sephadex LH-20 column chromatography (20 mmφ × 200 mm, manufactured by Pharmacia) using methanol as an elution solvent was performed on fraction B (dry solid 2.8 g). That is, the column was washed with 1.5 BV methanol and eluted with 0.5 BV methanol to obtain fraction B-1 (dried solid 1.16 g).
Further, fraction B-1 (1.16 g of the dried product) was subjected to flash ODS column chromatography (20 mmφ × 100 mm, manufactured by Nomura Chemical Co., Ltd.) using water / methanol as an elution solvent. That is, after washing the flash ODS column with 180 ml of 50% (v / v) methanol, stepwise wash sequentially with 60% (v / v) methanol, 70% (v / v) methanol, then 80 Elution with% (v / v) methanol gave fraction B-1-1 (319 mg of dry solid) containing the desired novel compound G.
Further, fraction B-1-1 (dry solid 319 mg) was fractionated by silica gel column chromatography (20 mmφ × 150 mm, manufactured by Kanto Chemical Co., Inc.) using hexane / ethyl acetate as an elution solvent. That is, after washing the silica gel column with 2 BV elution solvent hexane / ethyl acetate (9: 1), elution fraction E (dried product 114 mg) with 1 BV elution solvent hexane / ethyl acetate (8: 2) Got.
Further, fraction E (dried product: 114 mg) was subjected to preparative high performance liquid chromatography (ODS column, 20 mmφ × 250 mm, Nomura Chemical Co., mobile phase: 68% (v / v) methanol, detection: 254 nm UV monitor) to obtain a novel compound G (dried product 3.6 mg).

(2) Structural analysis of the new compound G The structural analysis of the new compound G was performed using high resolution mass spectrometry (HRFABMS) and nuclear magnetic resonance spectroscopy ( ¹ H NMR, ¹³ C-NNMR). The results are shown below.

(2-1) Results by high resolution mass spectrometry (HRFABMS) In high resolution mass spectrometry (HRFABMS), “m / z 399.1813 [M + H] ⁺ (calcd. 399.1808 Δ 0.5 mmu).” Was observed. The molecular formula was found to be “C ₂₃ H ₂₆ O ₆ ”.

(2-2) In the nuclear magnetic resonance spectroscopy ⁽¹ H NMR) Results from nuclear magnetic resonance spectroscopy ⁽¹ H NMR), the following peaks were observed.
^"1 H NMR (CDCl _3, 500 MHz): δ 0.99 (H-12 and H-13, 6H, s), 1.80 (H-8 ', 3H, s), 2.13 (H-7', 3H, s ), 2.34 (H-4 ', 2H, dd), 2.50 (H-5', 2H, t), 2.68 (H-9, 2H, s), 3.89 (H-11, 2H, s), 6.62 ( H-3 ', 1H, t), 7.67 (H-7, 1H, t), 7.74 (H-6, 1H, t), 8.07 (H-8, 1H, d), 8.09 (H-5, 1H , d).

(2-3) In the nuclear magnetic resonance spectroscopy ⁽¹³ C NMR) Results from nuclear magnetic resonance spectroscopy ⁽¹³ C NMR), the following peaks were observed.
^“13 C NMR (CDCl _3, 125 MHz): δ 12.3 (C-8 ′), 22.7 (C-4 ′), 25.2 (C-12 and C-13), 29.9 (C-7 ′), 32.1 ( C-9), 37.0 (C-10), 42.1 (C-5 '), 72.9 (C-11), 121.8 (C-3), 126.1 (C-8), 127.0 (C-5), 128.9 ( C-2 '), 129.4 (C-8a), 132.9 (C-4a), 133.0 (C-7), 135.0 (C-6), 140.0 (C-3'), 154.3 (C-2), 167.9 (C-1 '), 181.3 (C-1), 184.8 (C-4), 207.3 (C-6'). ''

  From the above results and the HMQC spectrum and HMBC spectrum, it was found that the new compound G was a new compound represented by the above formula (7).

1-6. Linacantin C (comparative example)
(1) Fractionation / isolation of linacantin C Fractionation / isolation of linacantin C was performed according to the flow shown in FIG. That is, dry roots (Rhinacanthus nasutus (L.) Kurz) (5 Kg) were extracted with 25 L 90% (v / v) ethanol three times at room temperature for 24 hours each. They were combined and concentrated to give a dried product (407 g). Next, this was suspended and dissolved in 7 L of 90% (v / v) methanol, then distributed three times with an equal amount of hexane, and then the 90% (v / v) methanol phase was taken and concentrated under reduced pressure. Purified water was added to the concentrate under reduced pressure, filled up to 5 L, transferred to a separatory funnel, and two-phase solvent partition was performed with chloroform three times. Next, the chloroform phases obtained by this operation were combined to obtain 69.3 g of a dried product.

Of this, 69.0 g was subjected to silica gel column chromatography (80 mmφ × 150 mm, manufactured by Kanto Chemical Co., Inc.) using hexane / ethyl acetate as an elution solvent. That is, linacantin C (dried product: 5.66 g) represented by the following formula (8) was obtained from the fraction eluted with 3 BV elution solvent hexane / ethyl acetate (9: 1).

When ¹ H NMR spectrum data (CDCl ₃ ) was obtained by nuclear magnetic resonance spectroscopy for linacantin C fractionated and isolated by the above method, the following peaks were observed, and the literature (Biol. Pharm. Bull Vol.27, 1070-1074 (2004)).
^"1 H NMR (CDCl _3, 500 MHz): δ 0.99 (H-12 and H-13,6H, s), 1.52 (H-8 ', 2H, d), 1.55 (H-10', 2H, s ), 1.76 (H-9 ', 2H, s), 1.98 (H-5', 2H, t), 2.13 (H-4 ', 2H, dd), 2.67 (H-9,2H, s), 3.88 (H-11,2H, s), 5.17 (H-7 ', 1H, dd), 6.66 (H-3', 1H, t), 7.64 (H-7,1H, t), 7.71 (H-6 , 1H, t), 8.04 (H-8,1H, d), 8.07 (H-5,1H, d). ''

2. Test example of antitumor action The test of antitumor action was performed by measuring the growth inhibitory activity against human-derived melanoma cell line HMV-II.

Human-derived melanoma cell line HMV-II was seeded in a 96-well culture plate (NUNC) at a cell concentration of 1 × 10 ⁴ cells / 90 μl using 10% (v / v) fetal bovine serum-containing ham F12 medium, 37 ° C. The culture was performed in the presence of 5% carbon dioxide for 24 hours. After 24 hours of culture, the novel compound A, novel compound B, novel compound D, novel compound F or novel compound G of the present invention (dissolved in DMSO, final DMSO concentration in the medium = 0.1% (v / v); DMSO The control group was cultured in a medium supplemented with 1/1000 volume of only, and further cultured at 37 ° C. in the presence of 5% carbon dioxide gas for 24 hours. Cell proliferation was measured by a method using MTT [3- (4,5-Dimethylthiazol-2-yl) -2,5-diphenyl tetrazolium bromide, Nacalai Tesque] [Tokyo Kagaku Doujin, published by Shinsei. Chemistry Experiment Course 12 Molecular Immunology I Immune cells / cytokines See pages 358-359. ].

That is, after 24 hours of culture was added each new compounds described above, 96 well culture plates 10 [mu] l of MTT solution to the culture solution 90 [mu] l of each well (0.33 cm ² / well) [5 mg / ml; calcium magnesium free It was dissolved in PBS (Dulbecco's Phosphate-Buffered Saline) solution, filtered through a membrane filter (0.22 μm)], shaken to homogenize, and cultured at 37 ° C. in the presence of 5% carbon dioxide for 4 hours. After 4 hours of culture, add 100 μl of 10% (w / v) SDS-50% (v / v) N, N-Dimethylformamide-0.005N hydrochloric acid solution to each well, and for 18 hours at 37 ° C with 5% carbon dioxide After standing still, using an immunoleader (manufactured by Dainippon Pharmaceutical Co., Ltd.), using 750 nm as a control, the absorbance at 590 nm was measured and used as an index of cell proliferation.

  Table 1 shows the cytostatic activity of each novel compound (novel compound A, novel compound B, novel compound D, novel compound F or novel compound G) against melanoma cell line HMV-II.

[Table 1]
Growth inhibition activity against human-derived melanoma cell line (HMV-II)
Concentration (μg / ml) Growth rate (%)
No addition (control example) 100
New Compound A 10 85
New compound B 10 74
New compound D 10 84
New compound F 10 84
New compound G 10 81
Linacantin C (Comparative Example) 10 86

  As shown in Table 1, each of the new compounds (new compound A, new compound B, new compound D, new compound F or new compound G) is equivalent to “human melanoma cells equivalent to linacantin C having excellent antitumor activity. It was confirmed that the strain has an excellent growth inhibitory activity against the strain (HMV-II).

Example 1. Preparation of tablets Tablets with the following formulation using novel compound A fractionated and isolated according to the method described in "1-1. Novel compound A (1) Fractionation and isolation of novel compound A" above (500 mg per tablet) is prepared.

New compound A 1mg
Lactose 479mg
10mg dried corn starch
Talc 9mg
Calcium stearate 1mg

(Preparation method)
Lactose (95.8 g) is mixed with novel compound A (0.2 g), dried corn starch (2 g), talc (1.8 g) and calcium stearate (0.2 g). Subsequently, a tablet is produced by a conventional method using a single-shot tablet press.

Example 2 Preparation of hard capsules Using the novel compound B fractionated and isolated according to the method described in “1-2. Novel compounds B and C (1) Fractionation and isolation of novel compounds B and C” above, Hard capsules (360 mg per capsule) are prepared with the following formulation.

New compound B 5mg
Lactose 220mg
Corn starch 110mg
Hydroxypropylcellulose 25mg

(Preparation method)
Lactose (220 g) and corn starch (110 g) are added to and mixed with the novel compound B (5 g), and an aqueous solution of hydroxypropylcellulose (25 g) is added thereto and kneaded. Next, granules are produced by an ordinary method using an extrusion granulator. Hard granules are prepared by filling the granules into gelatin hard capsules.

Example 3 Production of Soft Capsule Using New Compound C Fractionated / Isolated in accordance with the Method described in “1-2. Novel Compound B, C (1) Fractionation / Isolation of New Compound B, C” above, Soft capsules (170 mg per capsule) are prepared with the following formulation.

New compound C 0.5mg
Soybean oil 169.5mg

(Preparation method)
New compound C (0.5 g) is added to and mixed with soybean oil (169.5 g). Next, soft capsules are prepared by filling soft capsules using a rotary soybean automatic molding machine according to a conventional method.

Example 4 Preparation of pills Using novel compound D fractionated and isolated according to the method described in "1-3. Novel compounds D and E (1) Fractionation and isolation of novel compounds D and E" above, Make pills (100mg per capsule) with the following recipe.

New compound D 0.5mg
Morohaya powder 20.0mg
Starch 30.0mg
Molasses 20.0mg
Tea extract 15.0mg
Soy fiber 14.0mg
Shellac 0.5mg

(Preparation method)
After mixing the raw materials with the above blending, adding a suitable amount of water, producing a homogeneous kneaded product with a kneader, rolling the kneaded product, making a round shape using a round machine and drying it to produce a pill To do.

Example 5 FIG. Preparation of pills Using the novel compound E fractionated and isolated according to the method described in “1-3. Novel compounds D and E (1) Fractionation and isolation of novel compounds D and E” above, Make pills (100mg per capsule) with the following recipe.

New compound E 0.5mg
Morohaya powder 20.0mg
Starch 30.0mg
Molasses 20.0mg
Tea extract 15.0mg
Soy fiber 14.0mg
Shellac 0.5mg

(Preparation method)
After mixing the raw materials with the above blending, adding a suitable amount of water, producing a homogeneous kneaded product with a kneader, rolling the kneaded material obtained, rounding it using a round machine and drying it, thereby making a pill Is made.

Example 6 Preparation of ointment The ointment was formulated with the following formulation using the new compound F fractionated and isolated according to the method described in “1-4. New Compound F (1) Fractionation and Isolation of New Compound F” above. Is made.

(Oil phase component)
0.1 g of new compound F
White petrolatum 20.0g
Mineral oil 20.0g
Stearyl alcohol 5.0g
Steareth-2 3.0g
Propylparaben 0.1g
Natural vitamin E 0.1g
(Aqueous phase component)
1,3-butylene glycol 5.0 g
0.4 g of phenoxyethanol
Polysorbate 60 4.5g
Purified water
Total amount 100g

(Preparation method)
The oil phase component and the water phase component are each heated to 80 ° C. to be uniform, and the water phase is added to the oil phase with stirring, followed by emulsification and cooling to prepare an ointment.

Example 7 Preparation of Lotion Using the new compound G fractionated / isolated according to the method described in “1-5. New Compound G (1) Fractionation / Isolation of New Compound G” above, the lotion was prepared according to the following formulation. Is made.

(Oil phase component)
0.1 g of new compound G
Polyoxyethylene (60 mol) hydrogenated castor oil 2.0 g
1,3-butylene glycol 5.0 g
(Aqueous phase component)
Glycerin 5.0g
Phenoxyethanol 0.3g
Citric acid 0.1g
Sodium citrate 0.2g
Ethanol 8.0g
Purified water
Total amount 100g

(Preparation method)
A lotion is prepared by uniformly dissolving the oil phase component and the aqueous phase component and adding the oil phase to the aqueous phase with stirring.

As described above, the novel compound, the antitumor agent and the pharmaceutical agent, food or cosmetic having antitumor action of the present invention have been described based on the above embodiment, but the present invention is not limited to the above embodiment, The present invention can be implemented in various modes without departing from the gist thereof.

Claims (21)

A novel compound represented by the following formula (1).

A novel compound represented by the following formula (2).

A novel compound represented by the following formula (3).

A novel compound represented by the following formula (4).

A novel compound represented by the following formula (5).

A novel compound represented by the following formula (6).

A novel compound represented by the following formula (7).

  An antitumor agent comprising the novel compound according to claim 1.   An antitumor agent comprising the novel compound according to claim 2.   An antitumor agent comprising the novel compound according to claim 3.   An antitumor agent comprising the novel compound according to claim 4.   An antitumor agent comprising the novel compound according to claim 5.   An antitumor agent comprising the novel compound according to claim 6.   An antitumor agent comprising the novel compound according to claim 7.   A pharmaceutical, food or cosmetic having antitumor activity, comprising the novel compound according to claim 1.   A pharmaceutical, food or cosmetic having antitumor activity, comprising the novel compound according to claim 2.   A pharmaceutical, food or cosmetic having antitumor activity, comprising the novel compound according to claim 3.   A pharmaceutical, food or cosmetic having antitumor activity, comprising the novel compound according to claim 4.   A pharmaceutical, food or cosmetic having antitumor activity, comprising the novel compound according to claim 5.   A pharmaceutical, food or cosmetic having antitumor activity, comprising the novel compound according to claim 6.   A pharmaceutical, food or cosmetic comprising the novel compound according to claim 7 and having an antitumor effect.

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