JP2008150348A - New compound - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To isolate and identify a useful new compound contained in royal jelly. <P>SOLUTION: The compound is selected from the group consisting of the compound A: 4,10-dihydroxy-(E)-2-decenoic acid, the compound B: 4,9-dihydroxy-(E)-2-decenoic acid, the compound C: 3,9-dihydroxydecanoic acid and the compound D: 10-hydroxydecanoic acid 10-O-β-D-glucopyranoside or its salt. <P>COPYRIGHT: (C)2008,JPO&INPIT

Description

  The present invention relates to a novel compound or a salt thereof.

  Royal jelly, a bee product, is attracting attention as a food rich in amino acids, vitamins and minerals, including proteins, carbohydrates and lipids. As the biological activity of royal jelly, serum cholesterol and lipid lowering action, growth promoting action, blood pressure lowering action, insulin-like action, etc. are known, and antibacterial action and antitumor action have also been reported (for example, Patent Documents 1, 2, and 3). Based on these effects, royal jelly is being put into practical use as a raw material for health foods and functional foods for the purpose of preventing, improving or treating diseases.

Patent Document 1 discloses a royal jelly-derived decenoic acid derivative.
JP2005-306848 Townsend, G.M. F. et al. , 1959. Nature, 183, 1270 Townsend, G.M. F. et al. , 1959. Proc. Am. Assoc. Cancer Res. , 3, 69 Townsend, G.M. F. et al. , 1960. Cancer Res. , 20, 503

  However, many of the substances contained in royal jelly have not yet been isolated and identified, and it is believed that useful new compounds may be contained.

  Accordingly, an object of the present invention is to isolate and identify useful novel compounds contained in royal jelly.

  As a result of intensive studies to solve the above-mentioned problems, the present inventor has found that a novel compound is contained in an extract from royal jelly, and has completed the present invention.

  The present invention provides the following four compounds AD or salts thereof:

  ADVANTAGE OF THE INVENTION According to this invention, the novel compound which has the effect related to a nourishing tonic action, an antitumor action, and an antibacterial action, and its salt can be provided.

  The compound of the present invention can be obtained by either a method of chemically synthesizing or a method of isolating from natural royal jelly utilizing physical or chemical properties. These methods are not particularly limited, and conventionally known methods can be used. For example, in the case of isolation from natural royal jelly, the following percolation method can be used.

The percolation process, filled with freeze-dried powder of royal jelly in chromatographic tube, the step of extracting by adding CHCl ₃ gradually, CHCl ₃ in the extraction residue - CHCl ₃ was added to acetone - resulting acetone extract, compounds A~D The process of obtaining the compound represented by these is included.

  The salt of the compound of the present invention includes a salt of a COOH group. The salt is not particularly limited as long as it is physiologically acceptable, for example, alkali metal salts such as sodium, potassium and lithium; alkaline earth metal salts such as calcium, magnesium and barium; ammonium salts; triethylamine Preferred examples include organic amine salts such as triethanolamine; basic amino acid salts such as lysine and arginine. Moreover, you may use these salts in combination of 1 type, or 2 or more types. These salts can be easily produced from the compound by a conventional method.

After synthesizing or isolating the compound of the present invention and a salt thereof, it can be purified according to a method usually used by those skilled in the art. The purification method is not particularly limited, and for example, these purification steps exemplified by extraction, chromatograph, distillation and the like may be performed a plurality of times.

  Furthermore, the compounds of the present invention can be chemically synthesized using methods commonly used by those skilled in the art and, if necessary, can be purified through the purification steps described above.

  Moreover, the compound of this invention can also be obtained from fermentation by the microorganisms which have the production ability of (alpha) -hydroxy decenoic acid or (alpha) -hydroxy decanoic acid, for example. As the microorganism, a naturally occurring microorganism may be used, or a microorganism capable of producing a target compound by a known genetic manipulation technique may be used. When fermentation by microorganisms is used, after culturing the microorganisms, the culture solution is sterilized by treatment with activated carbon, filtration, etc., by precipitation with an organic solvent such as alcohol, desalting by ultrafiltration, etc. The compound of the present invention can be obtained by purification.

  The compound of the present invention can be expected to have an effect similar to that of 10-hydroxy-δ-2decenoic acid contained in natural royal jelly due to its chemical structure. That is, the compound of the present invention can be used in anticipation of effects related to nourishing tonicity, antitumor activity, and antibacterial activity. In addition, the salt of the compound can also be used in expectation of the above effect due to its chemical structure. Specific medicinal effects include nutritional tonic, post-disease, febrile debilitating disease, nutritional supplements such as nutritional disorders, and improvement of weak constitution.

  The compounds and salts thereof of the present invention can be applied to various uses for the purpose of the above-mentioned effects. Applications include, for example, food compositions such as foods, health foods, functional foods, supplements, foods for specific uses, foods for specific health use, pet foods, supplements for pets, therapeutic agents, reagents (for research and test kits) And toiletries (cosmetics and bath cosmetics).

  As a food composition, an antioxidant, a flavoring agent, a thickener, an emulsion stabilizer, an antiseptic, a flavoring agent, a sweetener, a sour agent, and the like and the compound of the present invention are treated according to a conventional method. Thus, the food composition of the present invention can be produced.

  The blending ratio of the compound or salt thereof blended in the food composition is not particularly limited as long as the expected effect of the compound or salt thereof can be obtained. It is about 0001-20 mg.

  When producing a food composition containing the compound, compounds A to D or fractions containing the compound obtained in the extraction process from royal jelly are added individually or in combination to the raw material of the food composition. Can do.

  When a fraction containing compounds A to D is blended in a food composition, the method for obtaining the fraction is not particularly limited as long as the desired effect as the novel compound of the present invention can be obtained. For example, royal jelly powder After extracting with chloroform, the residue is extracted with chloroform-acetone to obtain a residue. Further, a fat-soluble substance is removed from the extract obtained by extraction with methanol using an ODS column or the like, and concentrated.

  For toiletries, various forms such as creams, cosmetics, etc. can be formulated by arbitrarily combining with surfactants, oils, moisturizers, film forming agents, pigments, fragrances and the like that are generally used as cosmetic ingredients. Milky lotion, lotion, skin conditioner, base cosmetic, lipstick, lip balm, concealer, eye color, foundation, nail enamel, facial cleanser, pack agent, massage agent, cleansing agent, serum, body soap, etc. . In addition, by combining in combination with components such as surfactants, thickeners, oil gelling agents, metal oxides, pigments, and fragrances that are generally used as hair cosmetic ingredients, That is, shampoos, hair rinses, hair treatments, hair conditioners, packs, and the like can be used.

  The compounding ratio of the compound or salt thereof blended in toiletries is not particularly limited as long as the expected effect of the compound or salt thereof can be obtained, but is usually about 0.0001 to 10% by weight. Further, the compounds A to D of the present invention can be added to the toiletry raw material alone or in combination.

  When the compound of the present invention or a salt thereof is used as a pharmaceutical composition, the salt of the compound is preferably a pharmaceutically acceptable salt. Such salts include alkali metal salts such as sodium and potassium, alkaline earth metal salts such as calcium and magnesium, ammonium salts, organic amine salts such as triethylamine and triethanolamine, and bases such as lysine and arginine. Preferred amino acid salts and the like. Moreover, you may use these salts in combination of 1 type, or 2 or more types.

  The amount of intake when used as the pharmaceutical composition is usually about 0.0001 to 10 mg / kg per day per adult.

  The pharmaceutical composition containing the compound of the present invention or a salt thereof may contain an excipient, a carrier or an additive. The excipient, carrier and additive are not particularly limited as long as they are usually used and pharmaceutically acceptable, and the kind and composition thereof can be appropriately changed.

  Examples of excipients include sodium chloride and sodium citrate, and examples of carriers include sterilized water, physiological saline, and various buffer solutions. Additives include thickeners, buffers, preservatives, preservatives, and the like.

  The compound of the present invention is an orally safe substance contained in natural royal jelly, and even if ingested in excess of the above range, it does not adversely affect the body.

EXAMPLES Hereinafter, although an Example is given and this invention is demonstrated in more detail, it cannot be overemphasized that this invention is not limited by these Examples.
Example 1
Hereinafter, a method for extracting the royal jelly compounds A to D will be described. Throughout Examples 1 and 2, vacuum evaporation was performed using a rotary evaporator.
[Extraction of Extract Containing Compounds AD from Royal Jelly]
Royal jelly (1 kg) was sequentially extracted with CHCl ₃ (3 L) and CHCl ₃ -acetone (1: 1, ₃ L) by a percolation method, and the extracts were concentrated under reduced pressure to give CHCl ₃ extract (52.1 g), CHCl _{3, respectively.} -Acetone extract (36.1 g) was obtained.

Operation method: Royal jelly (1 kg) was packed in a chromatograph tube (inner diameter 8.0 × 45.0 cm), and CHCl ₃ was gradually added from above until the entire royal jelly was covered with the cock open. When the extract began to drop, the cock was closed and left at room temperature for 1 day, after which the extract was allowed to flow out at a rate of 3 mL per minute. Furthermore, CHCl ₃ (3 L in total) was added in a timely manner to continue the outflow, and the obtained extract was concentrated under reduced pressure to obtain CHCl ₃ extract (52.1 g). Next, CHCl ₃ -acetone (1: 1) was gradually added to the extraction residue from above until the entire extraction residue was covered, and the cock was closed when the extraction liquid began to drop. The extract was allowed to stand at room temperature for 1 day, and then the extract was allowed to flow out at a rate of 3 mL per minute. Thereafter, CHCl ₃ -acetone (1: 1, 3 L in total) was added in a timely manner to continue the outflow, and the obtained extract was concentrated under reduced pressure to obtain CHCl ₃ -acetone extract (36.1 g).

  The extraction method from royal jelly is shown in FIG.

Example 2
[Method for Isolating Compounds A to D]
The CHCl ₃ -acetone extract (36.1 g) obtained in Example 1 was subjected to silica gel chromatography (see separation condition 1), Sephadex LH-20 (see separation condition 2), ODS (separation) according to the procedure shown in FIG. Fraction (hereinafter referred to as fr.) 1 to fr. While monitoring the elution pattern of spots in normal phase thin layer chromatography and reverse phase thin layer chromatography. Fractionated into 4.

fr. 1 (68.8 mg) was purified by high performance liquid chromatography (see Separation Condition 4), Compound A (18.4 mg, retention time 43 minutes, yield 1.84 × 10 ⁻³ %), Compound B (1. 3 mg, retention time 36 minutes, yield 1.3 × 10 ⁻⁴ %).

fr. 2 (499.3 mg) was added CH ₃ CN (5 mL), and the dissolved portion was purified by high performance liquid chromatography (see Separation Conditions 5) to obtain Compound C (5.4 mg, retention time 26 minutes, yield 5.4). × 10 ⁻⁴ %) was obtained.

fr. 3 (42.6 mg) was purified by high performance liquid chromatography (see Separation Condition 6) to obtain Compound D (0.4 mg, retention time 19 minutes, yield 0.4 × 10 ⁻⁴ %).

The measurement results of Compound A are as follows.
- (1) oil, colorless, odorless - (2) Identification condition ^{Negative ion HR-FAB-MS m} / z: 201.1118 [M-H] - (Calcd for C 10 H 17 O 4: 201.1127)

¹ H-NMR (CD ₃ OD) δ:
_{1.14~1.58 (10H, m, C H} 2 × 5)
3.54 (2H, t, J = 6.6 Hz, H ₂ -10)
4.22 (1H, m, H-4)
5.97 (1H, dd, J = 1.8, 15.6 Hz, H-2)
6.92 (1H, dd, J = 4.8, 15.6 Hz, H-3)

¹³ C-NMR (CD ₃ OD) δ:
26.5, 26.9, 30.5, 33.6 (C-9), 37.6 (C-5), 63.0 (C-10), 71.6 (C-4), 120. 9 (C-2), 152.9 (C-3), 170.0 (C-1)

The measurement results of Compound B are as follows.
- (1) oil, colorless, odorless - (2) Identification condition ^{Negative ion HR-FAB-MS m} / z: 201.1117 [M-H] - (Calcd for C 10 H 17 O 4: 201.1127)

¹ H-NMR (CD ₃ OD) δ:
1.14 (3H, d, J = 6.6 Hz, H ₃ -10)
1.34 to 1.58 (8H, m, C H ₂ × 4)
3.72 (1H, m, H-9)
4.22 (1H, m, H-4)
5.97 (1H, d, J = 15.6 Hz, H-2)
6.89 (1H, dd, J = 4.8, 15.6 Hz, H-3)

¹³ C-NMR (CD ₃ OD) δ:
23.5 (C-10), 26.5, 26.8, 37.7 (C-5), 40.1 (C-8), 68.6 (C-9), 71.6 (C- 4), 121.7 (C-2), 152.2 (C-3), 170.6 (C-1)

The measurement results of Compound C are as follows.
- (1) amorphous, colorless, odorless - (2) Identification condition ^{Negative ion HR-FAB-MS m} / z: 203.1291 [M-H] - (Calcd for C 10 H 19 O 4: 203.1283)

¹ H-NMR (CD ₃ OD) δ:
1.14 (3H, d, J = 6.6 Hz, H ₃ -10)
_{1.34~1.48 (10H, m, C H} 2 × 5)
2.39 (1H, dd, J = 8.4, 15.0 Hz, H-2a)
2.48 (1H, dd, J = 4.8, 15.0 Hz, H-2b)
3.71 (1H, m, H-9)
3.97 (1H, m, H-3)

¹³ C-NMR (CD ₃ OD) δ:
23.5 (C-10), 26.7, 26.8, 30.7, 38.1 (C-4), 40.2 (C-8), 43.3 (C-2), 68. 6 (C-9), 69.4 (C-3), 174.1 (C-1)

The measurement results of Compound D are as follows.
- (1) amorphous, slightly yellow, odorless - (2) Identification condition ^{Negative ion HR-FAB-MS m} / z: 349.1869 [M-H] - (Calcd for C 16 H 29 O 8: 349.1862 )

¹ H-NMR (C ₅ D ₅ N) δ:
_{1.07~1.67 (14H, m, C H} 2 × 7)
2.32 (2H, t, J = 7.2 Hz, H ₂ -2)
3.68 (1H, m, H-10a)
3.99 (1H, m, H-5 ')
4.07 (1H, dd, J = 7.8, 7.8 Hz, H-2 ′)
4.11 (1H, m, H-10b)
4.24 to 4.29 (2H, m, H-3 ', 4')
4.41 (1H, dd, J = 4.8, 11.4 Hz, H-6′a)
4.59 (1H, dd, J = 2.4, 11.4 Hz, H-6′b)
4.87 (1H, d, J = 7.8 Hz, H-1 ′)

¹³ C-NMR (C ₅ D ₅ N) δ:
25.2, 26.4, 29.3, 29.6, 29.6, 29.6, 34.1, 34.9, 62.9 (C-6 '), 69.8 (C-10) 71.8 (C-4 '), 75.3 (C-2'), 78.6 (C-3 '), 78.6 (C-5'), 104.8 (C-1 ') 174.0 (C-1)

In this example, the equipment and reagents used for measuring the physical properties of compounds A to D are as follows.
[Measurement of nuclear magnetic resonance (NMR) spectrum]
It measured using JEOL JNM-GX400 and ECA-600SN nuclear magnetic resonance apparatus (JEOL Ltd.). The chemical shift value was expressed as a δ value (ppm) using tetramethylsilane (TMS) as an internal standard, and the coupling constant (J) was expressed in hertz (Hz). The following abbreviations were used for signal display. (S: singlet, d: doublelet, t: triplet, dd: double doublet, m: multiplet)

[Measurement of mass spectrometry (MS)]
Using a JEOL JMS-700T system (JEOL Ltd.), negative ion HR-FAB-MS was measured under the conditions of Gas: Xe and matrix: triehtanamine.

[High-performance liquid chromatography (HPLC)]
JASCO PU-2086 (JASCO Corporation) was used as a pump, and a GL Sciences 504R differential refractometer (GL Science Corporation) was used as a detector.

[Thin layer chromatography (TLC)]
The conditions of normal phase thin layer chromatography (normal phase TLC) are as follows.
Thin layer species: TLC aluminum sheets Silica gel 60 F ₂₅₄ (MERCK)
Developing solvent: CHCl ₃ -MeOH-H ₂ O 7: 3: 0.5
Detection method: 5% H ₂ SO ₄ / MeOH was sprayed and then colored by heating.

The conditions of reverse phase thin layer chromatography (reverse phase TLC) are as follows.
Thin layer type: TLC plates RP-18 F _254S (MERCK)
Developing solvent: 80% MeOH
Detection method: 5% H ₂ SO ₄ / MeOH was sprayed and then colored by heating.

The separation conditions for compounds A to D are as follows.
[Separation conditions]
Condition 1
Column type: silica gel # 7734 (MERCK)
Column dimensions: Internal diameter 5.0 x 60.0 cm (700 g)
Column temperature: 25 ° C (room temperature)
Solvent: CHCl ₃ -MeOH-H ₂ O 7: 3: 0.5 → 6: 4: 1
Flow rate: 10 mL / min

Condition 2
Column type: Sephadex LH-20 (Amersham Pharmacia Biotech)
Column dimensions: ID 2.0 × 45.0 cm (40 g)
Column temperature: 25 ° C (room temperature)
Solvent: 100% MeOH
Flow rate: 10 mL / min

Condition 3
Column type: Cosmosil 75C ₁₈ -OPN (Nacalai Tesque) for column chromatography
Column dimensions: ID 4.0 × 15.0 cm (80 g)
Column temperature: 25 ° C (room temperature)
Solvent: MeOH-H ₂ O 1: 9 → 3: 7 → 5: 5
→ 100% MeOH
Flow rate: 10 mL / min

Condition 4
Column type: Cat. No. 25415-96 Mightysil RP-18 GP 250-4.6 (5 μm) Cica-Reagent (Kanto Chemical Co., Inc.)
Column temperature: 25 ° C (room temperature)
Solvent: MeOH-Trifluoroacetic acid (hereinafter TFA) -H ₂ O 25: 0.5: 75
Flow rate: 0.5 mL / min
Detector: Differential refractometer

Condition 5
Column type: (Cat. No. 5020-04644 Inertsil ODS-3 4 μm 4.6 × 100 mm (GL Science Co., Ltd.)) × 3 Column temperature: 25 ° C. (room temperature)
_{Solvent: CH 3 CN-TFA-H} 2 O 25: 0.5: 75
Flow rate: 0.5 mL / min
Detector: Differential refractometer

Condition 6
Column type: (Cat. No. 5020-04644 Inertsil ODS-3 4 μm 4.6 × 100 mm (GL Science Co., Ltd.)) × 3 Column temperature: 25 ° C. (room temperature)
_{Solvent: CH 3 CN-TFA-H} 2 O 25: 0.5: 75
Flow rate: 0.6 mL / min
Detector: Differential refractometer

The extraction method from royal jelly is shown. The isolation method of compound AD is shown.

Claims (1)

Any compound or salt thereof selected from the group consisting of the following compounds A to D:

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