JP2003018974A - Food composition - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a food composition that has a variety of physiological activities, for example, the activation of energy metabolism and the immunostimulation. SOLUTION: The food composition includes a kind of non-pungent capsicin established as 'H-19 unpungent fixed strain', the dried (and crushed) product thereof and the extract from the (processed) plant bodies with ethyl acetate or ethanol.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a food composition containing "CH-19 sweet" which is a non-pungent fixed variety of pepper.

[0002]

BACKGROUND OF THE INVENTION Pepper (Capsicum annuum L. ) is a plant widely used worldwide as a food, spice and pharmaceutical ingredient. Its main pungent component is capsaicin ((E) -N-[(4-hydroxy-3-methoxyphenyl) -methyl] -8-me
thyl-6-nonenamide) and dihydrocapsaicin, that is,
It is a 6,7-dihydro derivative of capsaicin. Furthermore, as a pungent component, although small, nordihydrocapsaicin (a compound having one methylene group less than dihydrocapsaicin), homocapsaicin (a compound having one more methylene group than capsaicin), and homodihydrocapsaicin (a compound having more methylene groups than dihydrocapsaicin) 12 or more substances called capsaicinoids such as compounds having one more group).

Hitherto, it has been known that capsaicin has various physiological activities, such as promoting adrenaline secretion and enhancing fatty acid oxidation to thereby suppress obesity (Buck). , SH; Burks, T.
F., Pharmacol . Rev. , 1986, 38 , 179-226, Suzuki, T .;
Iwai, K., Chemistry and Pharmacology, 1984, Vol .2
3, 1984, Szolcsanyi, J., Handbook of Experimental Pha
rmacology, Vol .60, 1982). However, the amount of capsaicin used is limited due to its strong pungency and invasiveness, and its use as a food additive or pharmaceutical has been quite limited. In addition, pepper, which is a food material, has been used only in limited foods. On the other hand, it has been reported that some capsaicinoids having no pungency have a bioactive effect such as promoting secretion of catecholamine from the adrenal gland like capsaicin (Watanabe, T .; Kawad
a, T .; Kato, T .; Harada, T .; Iwai, K., Life Sci .,
1994, 54, 369-374). Since these capsaicinoids have no pungency, they may be used as food additives or pharmaceuticals.

[0004]

Problems to be Solved by the Invention A spicy fixed variety obtained in Thailand, "CH-19" (Kyoto Prefectural University, Faculty of Agriculture, Vegetable Horticulture Laboratory introduction number) was selected and fixed in an experimental field of Kyoto University. It is reported that capsicinoid is hardly contained in "CH-19 sweet", a non-pungent fixed variety of pepper, and that it contains a large amount of capsaicinoid-like substances (Yazawa, S .; Suetome, N. .;
Okamoto, K .; Namiki, T., J. Japan Soc . Hort . Sci. ,
1989, 58 , 601-607). The present inventors have determined that this “C
The structural analysis of the capsaicinoid-like substances contained in "H-19 sweet" was attempted, and these substances were successfully identified for the first time, and the present invention was completed.

[0005]

That is, the present invention provides a compound represented by the following general formula:

[0006]

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[0007]

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[0008] The present invention relates to a novel capsaicinoid-like substance represented by any of the above. Whereas in conventional capsaicinoids, vanillyl alcohol and branched unsaturated fatty acids have an amide bond, whereas the substance shown in Chemical formula 1 of the present invention has an ester bond between vanillyl alcohol and branched unsaturated fatty acids. There is a feature. The chemical substance 2 is a substance in which the fatty acid portion of the chemical substance 1 is saturated. In these chemical formulas, n is preferably 3, 4 or 5. For example, the structural formula of a substance in which n is 4 is represented by the following chemical formulas 3 and 4.

[0009]

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[0010]

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The capsaicinoid-like substance of the present invention is used as a food additive or a pharmaceutical ingredient having various physiologically active actions such as activation of energy metabolism and activation of immunity since it has no pungency and substantially no cytotoxicity. it can. Furthermore, it may be used as a food additive or pharmaceutical ingredient for maintaining durability in continuous exercise, and as a non-narcotic analgesic. Accordingly, the present invention also relates to a food composition containing the capsaicinoid-like substance. The food composition of the present invention can take any form such as solid, liquid, sol, gel, powder and granule, and can be produced by any production method known in the art. Those skilled in the art can appropriately determine the amount of the capsaicinoid-like substance of the present invention contained in the food composition according to the purpose of blending, the type, form, purpose, and the like of the food composition. For example, there is chocolate or the like containing at least ^10-5 % by weight or more of a capsaicinoid-like substance. The capsaicinoid-like substance of the present invention contained in the food composition does not necessarily have to be purified. For example, "CH-19 sweet" itself, a non-pungent fixed variety of hot pepper (unprocessed product), its dried product (crushed product), or alcohols represented by ethyl acetate, ethanol, and food emulsifiers, etc. It may be contained in the food composition as an extract of “CH-19 sweet” with various solvents commonly used for extraction from natural products in the technical field.

Furthermore, the present invention also relates to a pharmaceutical composition containing the above capsaicinoid-like substance. The pharmaceutical compositions of the present invention can take any form known in the art, for example, liquid formulations such as solutions, suspensions, and emulsions buffered with various salts and buffers. be able to. Salts can be alkali and alkaline earth metal salts, phosphates and sulfates. Various buffers, eg, citrate, phosphate, HEPES, Tris, etc., can be used at physiologically acceptable concentrations. The pharmaceutical composition of the present invention can be in the form of tablets, powders, sols, gels, granules, and liposomes, in addition to liquid preparations. In preparing various formulations, various excipients or additives known to those skilled in the art can be used in pharmaceutically acceptable amounts. The amount of the capsaicinoid-like substance of the present invention contained in the pharmaceutical composition can be appropriately determined by those skilled in the art according to the properties of other components, the purpose of use, the age and weight of the patient, the required effects, and the like. The pharmaceutical composition can be administered by various routes of administration, for example, oral, intravenous, subcutaneous, intramuscular, intraperitoneal, and nasopharyngeal, depending on the form and formulation.

[0013]

BEST MODE FOR CARRYING OUT THE INVENTION The capsaicinoid-like substance of the present invention is produced by separating and purifying from a non-pungent variety of pepper, "CH-19 sweet", as shown in Examples described later. Those skilled in the art can easily synthesize chemically using known reaction techniques based on the structural formulas described in this specification. For example, the compound of the present invention represented by Chemical formula 4 can be easily synthesized chemically by a well-known esterification reaction using 8-methylnonanoic acid (8-methylpelargonic acid) and vanillyl alcohol as starting materials. Can be. Hereinafter, the present invention will be described in more detail with reference to examples, but the examples do not limit the present invention in any way.

[0014]

【Example】Example 1 of the capsaicinoid-like substance of the present invention
Separation / purification Freeze-dry 1.00 kg of fresh fruit of "CH-19 sweet"
After removing seeds and calyx,
1.8 L each time using a
Extracted with chill. The obtained ethyl acetate extract was removed under reduced pressure.
Distill to remove the ethyl acetate, oleoresin (resin oil
7.3 g). This oleoresin is converted to silica gel (S
(ilica gel 60 Merck; 36 x 200 mm)
And eluted stepwise with n-hexane and ethyl acetate
I let it. The fraction eluted with ethyl acetate was separated on silica gel (Wakos
reversed-phase chromatography on il 25C18 Wako Pure Chemicals; 20 x 50 mm)
Raph and elute with 75% methanol
(3) 4.5 mg was obtained. On the other hand, n-hexane / acetic acid
The fraction eluted with chill (80:20) was collected on silica gel (Wakos
reversed phase chromatography on il 25C18 Wako Pure Chemicals; 20 x 90 mm)
Raph and elute with 75% methanol
A mixture containing (1) was obtained. Similarly, from the next fraction
As a result, 59.7 mg of compound (2) was obtained as a colorless oil.
The mixture containing compound (1) was mixed with silica gel (Wakosil 2
5C18 Wako Pure Chemicals; 20 x 90 mm) on reverse phase chromatography
And 75% methanol (0.05M AgNO₃
Contained). The resulting eluate is CHCl₃Three times
Extract and collect CHCl3 fractions to dry NaCl₂SO₄To
And then filtered. The filtrate is evaporated under reduced pressure
98.5 mg of compound (1) was obtained as a colorless oil. In addition,
In the separation / purification procedure of the compound, compound (1) and
And compound (2) as a developing solvent on thin-layer chromatography.
As toluene: chloroform: acetone (55: 2
6:19 v / v) and 0.1% 2,6-dichloroquinone
-4- Development of chloroimide (dissolved in 85% methanol) solution
Sprayed on the thin layer plate, and
It was tracked and confirmed by the blue color resulting from the reaction. Also
Compound (3) is toluene: chloroform:
Acetone (29:32:39 v / v) was used.

[0015]Example 2: Capsaicinoid-like substance of the present invention
Structure determination The instruments used for the spectroscopic analysis of the compounds of the present invention are as follows
Normal operation according to the respective operation manual
Was done.¹ H-NMR (399.65 MHz, CDCl₃)as well as^ThirteenC-NM
R (100.40 MHz, CDCl ₃) Spectrum (Internal standard: TM
S): JEOL alpha-400, IR spectrum: day
Standing 270-50 infrared spectrophotometer, UV spectrum: Ja
sco UVIDEC 660 spectrophotometer, HRMS
Vector: JEOL JMS-700.

[0016]Structure determination of compound (1):The above measuring means
The spectrum data of compound (1) obtained by
It is as follows.HRMS m / z (M ⁺ ) : Calculated value (C₁₈H₂₆O₄): 306.1831 Measurement value: 306.1798IRνmax (film) cm ^-1 : 3450, 1740, 1
615, 1610, 1520, 1470, 1435, 1
275, 1160, 1120, 1035, 970, 85
0,815,795,560.UVmax (MeOH) nm : 280 (ε: 2400), 23
1 (ε: 6200) ¹ H and ¹³ C-NMR spectra The results are shown in Tables 1 and 2, respectively.

From the results of the above HRMS, compound (1)
Was determined to be C ₁₈ H ₂₆ O ₄ . In the IR spectrum, absorption by hydroxy (3450 cm
^-1 ) and absorption by ester carbonyl (1740c
m- ¹ ) was found. ^{The 1} H-NMR spectrum shows three aromatic protons (δ 6.90d, 6.86d, and 6.87d), and these three coupling constants and patterns are typical for 1,2,4-substituted A phenyl group was indicated. By this phenyl group, ¹³ C-NM
In the R spectrum, δ146.5, 145.8, 1
Signals of 28.0, 122.0, 114.3 and 111.2 were seen. 2 coupled to each other at 15.6 Hz seen in the ¹ H-NMR spectrum
Two olefinic methine protons (δ 5.37 dd and 5.30 dt) indicated the presence of an ethylene moiety in the trans configuration. Also, a methoxy group (δ 3.90s) and an isopropyl group (δ 0.95d, 0.95d and 2.
21 oct) was observed.

As is clear from the data in Table 2, the ¹³ C-NMR spectrum of compound (1) is very similar to capsaicin (genuine product), and only methylene carbon (C-7 ′) is δ43 in capsaicin. Whereas the compound (1) had a difference of δ66.3. Both are similar for the ¹ H-NMR spectrum. However, the chemical shift value (δ 5.03, s) of the methylene proton of compound (1) was different from the methylene of carbon (C-7 ′) of capsaicin. The NMR signals of these methylenes in compound (1) indicate the presence of a methylene group sandwiched between the oxygen atom of the ester bond and the phenyl group. Therefore, it was presumed that the compound (1) had an ester moiety instead of the amide moiety of capsaicin. Also, ¹ H-N related to the acyl group
Since the MR spectrum and the ¹³ C-NMR spectrum of the compound (1) and capsaicin agree very well, the compound (1) has the same acyl group as capsaicin, that is, (E) -8-methyl-6- It was presumed to have a nonenoyl group. From the above data, the structure of compound (1) of the present invention was determined to be 4-hydroxy-3-methoxybenzyl (E) -8-methyl-6-nonenoate shown in Chemical formula 3, and "capsiate" It was named.

[0019]Structure determination of compound (2):The above measuring means
The spectrum data of compound (2) obtained by
It is as follows.HRMS m / z (M ⁺ ) : Calculated value (C₁₈H₂₈O₄): 308.1987 Measurement value: 308.2008IRνmax (film) cm ^-1 : 3450, 1740, 1
615, 1610, 1520, 1470, 1435, 1
275, 1160, 1120, 1035, 970, 85
0,815,795,560.UVmax (MeOH) nm : 279 (ε: 3700), 23
1 (ε: 8700) ¹ H and ¹³ C-NMR spectra The results are shown in Tables 1 and 2, respectively.

From the results of the above HRMS, compound (2)
Was determined to be C ₁₈ H ₂₈ O ₄ . The IR spectrum of compound (2) was consistent with that of compound (1). Therefore, it is presumed that the structure of compound (2) is similar to the structure of compound (1). Further, ¹ H-NMR spectrum and ¹³ C-NMR spectrum data of the compound (2) were similar to those of the compound (1). However, instead of the two olefinic methine carbons (C-6 and C-7) found in compound (1),
Two alkane methine carbons (δ 27.2 and 38.
The presence of 9) was observed. Also in the data of the ¹ H-NMR spectrum, the signal of the olefinic proton as observed in the compound (1) was not recognized. From the above results, the structure of the compound (2) of the present invention was
6,7-dihydro derivative of compound (1), that is, 4-hydroxy-3-methoxybenzyl 8
-Methyl nonanoate.

Chemical synthesis of compound (2): A mixture of 500 mg (2.9 mmol) of 8-methylnonanoic acid and 3.5 g (29 mmol) of thionyl chloride was stirred overnight at room temperature using a magnetic stirrer under drying with CaCl _2. did. It was then evaporated under reduced pressure to give a brown oil. 893 mg of vanillyl alcohol (5.8
mmol) containing 5 mL of a pyridine solution.
The mixture was stirred with a magnetic stirrer at 2 ° C. for 2 hours.
Thereafter, water and 2N-HCl were added to this to make it acidic,
The resulting mixture was extracted with 30 mL of ethyl acetate each time. These ethyl acetate fractions were collected, washed with water, dried over anhydrous Na ₂ SO ₄ and filtered. The filtrate was evaporated under reduced pressure to give a residue. This residue is converted to silica gel (Silic
agel 60 Merck; 36 × 60 mm), and 2.329 mg of a colorless oil (yield: 3) was obtained from the fraction eluted with n-hexane and ethyl acetate (90:10).
6.8%). The data of the IR spectrum, ¹ H-NMR spectrum and ¹³ C-NMR spectrum of the compound thus synthesized were completely identical with those of the compound (2). Therefore, it was confirmed that the compound (2) was 4-hydroxy-3-methoxybenzyl 8-methylnonanoate, and was named “dihydrocapsiate”. Compound (3) was identified as vanillyl alcohol as a result of comparing the data of the above various spectra with those of the pure reagent.

[0022]

[Table 1]

[0023]

[Table 2]

[0024]Example 3: Capsaicinoid-like substance of the present invention
Stimulatory effect of immunity The immunostimulatory effect of the capsaicinoid-like substance of the present invention was examined.
To discuss the effect of hybridomas on antibody production.
Examined. The capsaicinoid of the present invention obtained in Example 1
Add 66μl of ethyl acetate to the like substance (20.5mg)
Then, a 1M solution was prepared. Next, 8 μl of this solution was added to IT
ES ERDF (Insulin (5 μg / ml)-Tiger
Nsferrin (35 μg / ml) -ethanolamine
ERD with (20 μM) -selenium (2.5 nM)
F) Dissolved in serum-free medium (40 ml), 200 μM solution
Was prepared. Using this solution, various concentrations of capsaicin
Prepare a medium containing the noid-like substance and prepare the following hybrid
Maize cell culture was performed. As a comparison, Capsaisi
A medium supplemented with an enzyme was also used.

[0025]

[Table 3]     Hybridoma antibody production       H8 human monoclonal antibody (IgM)       S97 Human monoclonal antibody (IgG)

The results obtained are shown in FIGS. 1, 2 and 3. From these results, the capsaicinoid-like substance of the present invention (denoted as “CLS-B” in each figure)
Compared to capsaicin, IgM and I of hybridoma
It turns out that gG antibody production is enhanced more effectively.
Furthermore, as is clear from FIG. 3, it is considered that the capsaicinoid-like substance of the present invention has extremely low cytotoxicity as compared with capsaicin.

[0027]Example 4: Capsaicinoid-like substance of the present invention
Activation of energy metabolism Next, the energy metabolism of the capsaicinoid-like substance of the present invention
Was examined for its activating effect. Dry grinding of "CH-19 sweet"
(Approximately 3 g) is wrapped in a medical wafer and eaten by the subject.
Let me know. The result is shown in FIG. It is clear from FIG.
As described above, the skin temperature rises 1.4 ° C about 30 minutes after ingestion
did. On the other hand, the temperature of the inner ear (eardrum) is about 75 minutes after ingestion
The temperature gradually decreased by about 0.3 ° C. In addition, it is a cultivated variety plant of pungent taste
Unlike a certain hawk's claw, in this experiment
No sluggishness was observed in any of the subjects. These results
The dry and crushed product of "CH-19 sweet" does not cause irritation.
To increase body temperature (activate energy metabolism)
). Further, the results obtained in Example 1 were obtained.
The capsaicinoid-like substance of the present invention was administered to mice.
The effect of activating energy metabolism was observed. Got
FIG. 5 shows the results. From these results, the capsaicin of the present invention
The body temperature of the mice receiving the noid-like substance was not
It is clearly higher than
Was.

Example 5: Capsaicinoid-like substance of the present invention
A food composition containing the capsaicinoid-like substance of the present invention was produced by a method known to those skilled in the art. The units of numerical values in the following examples are all parts by weight.

[0029]

[Table 4]

[0030]

[Table 5] Sports drinks : Orange concentrate juice 0.200 Sugar 1.8 Isomerized sugar (F-55) 5.5 Citric acid 0.14 Salt 0.08 Sodium citrate 0.07 Potassium chloride 0.04 Daiichi Calcium phosphate 0.013 Sodium glutamate 0.004 Magnesium chloride 0.003 Ascorbic acid 0.1 Cloudy 0.1 Emulsified perfume 0.01 Essence 0.2 Capsaicinoid-like substance (*) 0.05 The balance (Water)

The capsaicinoid-like substance (*) used in the above-mentioned sports drink was prepared by freeze-drying 1.00 kg of fresh fruit of "CH-19 sweet" to remove seeds and calyx, and then using a universal homogenizer (Japan). It is oleoresin (resin oil) obtained by extracting with 1.5 L of ethyl alcohol each time using a Seiki Seisakusho and evaporating the obtained extract under reduced pressure. The capsaicinoid-like substance of the present invention is about 50 ppm. It contains.

[0032]

[Table 6] Royal jelly beverage : Isomerized sugar (F-55) 12.0 Purified honey 11.0 Raw royal jelly 4.5 Garlic extract 0.2 Litchi extract 0.3 Citric acid 0.1 Polydextrose 4.0 Natural cafe In 0.08 Ascorbic acid 0.5 Vitamin B1 hydrochloric acid 0.02 Vitamin B2 phosphate 0.01 Vitamin B6 Hydrochloric acid 0.03 Nicotinamide 0.04 Cloudy 0.1 Essence 0.4 Capsaicinoid-like substance / Or the substance of Chemical formula 4) 0.001 balance (water)

[0033]

[Table 7]Retort-Red bean porridge with paraty Brown rice Koshihikari 4.20 White rice Koshihikari 4.80 Hokkaido red beans 1.60 Sugar 0.50 Salt 0.10 "CH-19 Sweet" Fruit 2.0 The rest (water)

The red beans were sorted out, immersed in water overnight and weighed. Fresh fruits of "CH-19 sweet" were immersed in boiling water for 5 minutes, and after blanching, cut into pieces of about 5 mm in size. After filling these and other components into an aluminum pouch bag containing 300 g so that the total amount becomes 300 g,
The mixture was retorted for 5 minutes. In addition, when the same experiment as Example 4 was performed using this retort food, an increase in body surface temperature was observed, and it was confirmed that “CH-19 sweet” itself also has an activating effect on energy metabolism. .

[0035]

As described above, a novel capsaicinoid-like substance having an ester bond was obtained. Unlike conventional capsaicin, this substance has no pungency and has substantially no cytotoxicity, so that it can be widely used as a food ingredient. In addition, it was confirmed that the capsaicinoid-like substance had an activating effect on immunity and an activating effect on energy metabolism, and was also found to have various physiological activating effects such as an anti-fatigue effect and an anti-obesity effect. It is useful as a food additive and a pharmaceutical ingredient.

[Brief description of the drawings]

FIG. 1 shows a capsaicinoid-like substance of the present invention (in each figure,
"CLS-B") and psycin
1 shows the effect of hybridoma H8 on IgM antibody production. In the figure, △ and ▲ are each 100μ
The values obtained when the cells were cultured in a medium supplemented with M and 20 μM CLS-B are shown. □ and △ are 100μ each
The values obtained when the cells were cultured in a medium supplemented with M and 20 μM capsaicin are shown.

FIG. 2 shows the capsaicinoid-like substance of the present invention (in each figure,
"CLS-B") and psycin
1 shows the effect of hybridoma S97 on IgG antibody production. In the figure, △ and ▲ are 100
The values obtained when the cells were cultured in a medium supplemented with μM and 20 μM CLS-B are shown. □ and △ are 100
The values obtained when the cells were cultured in a medium to which μM and 20 μM capsaicin were added are shown.

FIG. 3 shows a capsaicinoid-like substance of the present invention (in each figure,
"CLS-B") and psycin
FIG. 4 shows the effect of the hybridoma S97 on the viability. In the figure, △ and ▲ are each 1
The values obtained when the cells were cultured in a medium supplemented with 00 μM and 20 μM CLS-B are shown. □ and △ are each 1
The values obtained when the cells were cultured in a medium supplemented with 00 μM and 20 μM capsaicin are shown.

FIG. 4 shows changes in the temperature of the skin and inner ear (eardrum) due to the ingestion of a dried and ground product of “CH-19 sweet”. The vertical axis indicates temperature change, and the horizontal axis indicates elapsed time (minutes) after ingestion.

FIG. 5 shows the capsaicinoid-like substance of the present invention weighing 1 kg.
5 shows the results of examining changes in body temperature after administration to mice at a ratio of 50 mg.

   ────────────────────────────────────────────────── ─── Continuation of front page    (72) Inventor Kenji Furuhata             21-11-203 Kawahara-cho, Shimizu-shi, Shizuoka (72) Inventor Masatake Imai             2-1-1 Shimosueyoshi, Tsurumi-ku, Yokohama-shi, Kanagawa             Morinaga & Co., Ltd. (72) Inventor Yuko Setoguchi             2-1-1 Shimosueyoshi, Tsurumi-ku, Yokohama-shi, Kanagawa             Morinaga & Co., Ltd. (72) Inventor Shuichi Hashizume             2-1-1 Shimosueyoshi, Tsurumi-ku, Yokohama-shi, Kanagawa             Morinaga & Co., Ltd. F term (reference) 4B018 MD61 ME02 ME14 MF01                 4C088 AB50 AC04 BA07 BA08 NA14                       ZB09 ZC21

Claims (3)

[Claims] 1. "C", a non-pungent fixed variety of pepper.
H-19 sweet ", a dried (ground) product thereof, or a food composition containing the ethyl acetate or ethanol extract thereof. 2. The food composition according to claim 1, wherein the food composition is an immune activated food. 3. The food composition according to claim 1, wherein the food composition is an activated food of energy metabolism.