JP4849388B2 - Physiologically active substance and method for producing the same, tasting method of pungent physiologically active substance, and health / medical composition - Google Patents

Description

  The present invention relates to a physiologically active substance and a method for producing the same, a method for tasting a pungent physiologically active substance, and a health / medical composition.

  More specifically, the invention of bioactive substances as labdan-type diterpene compounds newly discovered from newly derived plants, the invention of the production method of these bioactive substances, and the known pungent bioactive substances with tasteless physiological activity The present invention relates to an invention of a taste-eliminating method for a pungent physiologically active substance to be converted into a substance, and an invention of a health / medical composition constituted using these physiologically active substances.

  Conventionally, ginger family plants such as ginger and turmeric have been consumed as spices and flavored vegetables, and are widely used in folk medicine to improve symptoms such as inflammation and gastrointestinal disorders. Yes.

  Furthermore, various labdan-type diterpene compounds derived from ginger and the like have been reported, and these compounds have various useful physiology such as antibacterial action, platelet aggregation inhibitory action, human 5-lipoxygenase inhibitory action, and anticancer action. It has been proposed to be used for antibacterial agents, antithrombotic agents, anti-inflammatory agents, anticancer agents and the like as having an active action. Moreover, since many of these labdan-type diterpene compounds exhibit a pungent taste, use as a taste component has also been proposed.

JP-A-63-162644 discloses (E) -8,17-epoxy-labd-12-ene-15,16-dial and other labdane-type diterpene compounds, and these compounds are disclosed in JP-A-63-162644. Are derived from Alpinia genus plants, have a pungent taste, and are expected to be useful as antitumor agents.

JP-A-6-25214 discloses the 7-aminolabdans and the production method thereof, and their use as a pharmaceutical preparation. The 7-aminolabdans exhibit blood pressure action and potassium channel pore activity. It is said to be effective for cardiovascular diseases, metabolic diseases, cancer chemotherapy and the like.

JP-A-2002-47195 discloses an anti-inflammatory pharmaceutical composition and an anti-platelet aggregation pharmaceutical composition obtained through a predetermined extraction operation, although the active ingredient is not specifically specified in Patent Document 3 above. Or an antifungal pharmaceutical composition is disclosed.

  On the other hand, Zingiber mioga, a kind of ginger family plant, is a unique herb in Japan and analyzes its active ingredients even though its sprouts and buds have been widely used as edible and herbal medicines since ancient times. There are few attempts to do so, and the reports are very limited.

  In such a situation, the present inventors have already described in the following Non-Patent Document 1 that Myoga has a labdan-type diterpene dialdehyde compound, miogadial, that is, (E) -8β (17). -Epoxylabd-12-ene-15,16-dial [(E) -8β (17) -epoxylabd-12-ene-15,16-dial], two kinds of acetal cyclized labdane diterpene aldehyde compounds, etc. It is clarified that the ingredient is included. In addition, the following non-patent document 2 reports that the components of myoga exhibit strong antibacterial activity, and that the components of myoga exhibit excellent apoptosis-inducing activity against cancer cells and strong platelet aggregation inhibitory activity are also described below. Non-Patent Document 3 and the like are reported.

M. Abe et al., Biosci. Biotech. Biochem., 66, 2698-2700 (2002) M. Abe et al., Biosci. Biotech. Biochem., 68 (7), 1601-1604 (2004) N. Miyoshi et al., Cancer Lett., 199, 113-119 (2003)

  By the way, the active ingredient of the ginger reported by the inventors of the present invention by the above Non-Patent Document 1 to Non-Patent Document 3 is a compound that has already been reported to exist in other ginger family plants and the like. In consideration of this point, the inventor of the present application reported various and powerful physiological activities peculiar to myoga reported in the above Non-Patent Document 1 to Non-Patent Document 3 etc. It is difficult to logically agree that it is carried only by myogadial or acetal cyclized labdane diterpene aldehyde compounds. The inventor of the present application has considered that myouga may contain various unknown and potent physiological activities from such considerations.

  Therefore, the present invention is a novel and capable of utilizing the characteristics of the physiologically active substance that has been identified by isolating an unknown powerful physiologically active substance that can be presumed to be present in myoga, elucidating its structure, and characterizing it. The development of useful usage is a technical issue to be solved.

(Configuration of the first invention)
The configuration of the first invention of the present application for solving the above problems is a labdan-type diterpene trialdehyde compound, and has at least one physiological activity of antibacterial action, platelet aggregation inhibitory action, or human 5-lipoxygenase inhibitory action , A physiologically active substance.

(Configuration of the second invention)
The configuration of the second invention of the present application for solving the above-mentioned problems is a physiologically active substance in which the labdane-type diterpene trialdehyde compound according to the first invention is a 15,16,17-trial compound having a labdane skeleton structure. .

(Configuration of the third invention)
The structure of the third invention of the present application for solving the above problem is that the 15,16,17-trial compound having a labdane skeleton structure according to the second invention is a tasteless compound represented by the general formula of “Chemical Formula 2” below. It is a physiologically active substance.

(In the above “Chemical Formula 2”, the atomic groups represented by position numbers 18, 19, and 20 are each a methyl group.)
(Configuration of the fourth invention)
The fourth aspect of the present invention for solving the above-described problem is that a physiologically active substance according to any one of the first to third aspects of the invention is collected from an extract of Zingiber mioga which is a ginger family plant. It is a manufacturing method of a substance.

(Structure of the fifth invention)
The configuration of the fifth invention of the present application for solving the above-mentioned problem is a method for producing a physiologically active substance, wherein the extract according to the fourth invention is prepared by hexane extraction of myoga.

(Structure of the sixth invention)
The configuration of the sixth invention of the present application for solving the above-described problem is that a crude enzyme extract prepared from a myoga flower is allowed to act on a raw material liquid containing a labdane-type diterpene aldehyde compound having an epoxy group bonded to an aromatic ring. It is the manufacturing method of the bioactive substance which produces | generates the bioactive substance which concerns on either of 1st invention-3rd invention, and extract | collects this.

(Structure of the seventh invention)
The composition of the seventh invention of the present application for solving the above-mentioned problem is that a crude enzyme extract prepared from a myoga flower bud is prepared with respect to a raw material solution containing a pungent labdane-type diterpene aldehyde compound having an epoxy group bonded to an aromatic ring It is a tasting method of a pungent physiologically active substance by converting the labdane-type diterpene aldehyde compound having the pungent taste into a tasteless physiologically active substance according to the third invention while maintaining its physiological activity.

(Configuration of the eighth invention)
The structure of the eighth invention of the present application for solving the above-described problem includes, as an active ingredient, at least a physiologically active substance according to any one of the first to third inventions, a crushed ginger or a ginger extract. Composition.

(Structure of the ninth invention)
The structure of the ninth invention of the present application for solving the above-mentioned problem is a health and medical composition in which the Myoga extract according to the eighth invention is a hexane extract of Myoga.

(Configuration of the tenth invention)
The structure of the tenth invention of the present application for solving the above-described problem is that the health / medical composition according to the eighth or ninth invention is an antibacterial agent, an antithrombotic agent or an anti-inflammatory agent. It is a composition.

(Structure of 11th invention)
The constitution of the eleventh invention of the present application for solving the above-mentioned problems is a health / medical composition that contains the physiologically active substance according to the third invention as an active ingredient and is taken orally.

(Effect of the first invention)
The inventor of the present application succeeded in isolating the physiologically active substance according to the first invention by precise compound search by combining various chromatographic operations described below in detail. It has already been confirmed in vivo or in vitro that this physiologically active substance exhibits an excellent antibacterial action, platelet aggregation inhibitory action, and human 5-lipoxygenase inhibitory action. Further, the inventor of the present application estimates that the physiologically active substance according to the first invention will further exhibit at least excellent renal function improving action and anticancer action. That is, the physiologically active substance of the first invention exhibits a great variety of physiological activities.

  Moreover, the physiologically active substance according to the first invention is a kind of labdan type diterpene compound, but not only greatly differs from the conventionally known labdane type diterpene compound in that it is a “trialdehyde” compound. It is a novel compound that is very rare in nature.

  It is considered that the rare structural feature of being a trialdehyde compound in the physiologically active substance according to the first invention is related firstly to the diverse and strong physiological activities of this physiologically active substance. Secondly, as a general theory, it is an important lead for the development of various pharmaceuticals and health food ingredients in the future in the sense that the trialdehyde structure of the compound provides valuable suggestions on the functions that can be shown for human physiology. Can be a compound.

(Effect of the second invention)
The physiologically active substance according to the first invention is more preferably a 15,16,17-trial compound having a labdane skeleton structure.

(Effect of the third invention)
The 15,16,17-trial compound having a labdane skeleton structure according to the second invention is more preferably a compound represented by the general formula of “Chemical Formula 2”.

  The inventor of the present application has confirmed that at least the physiologically active substance according to the third invention is a tasteless compound. This point is in sharp contrast with the point that myogadiar, which is a 15,16-diar compound having a labdane skeleton structure, which is a closely related compound, exhibits a pungent taste.

  Therefore, firstly, the physiologically active substance according to the third aspect of the present invention can be considered for use as a health / medical composition for oral intake without pungent taste. Secondly, in terms of the difference between one aldehyde group in closely related compounds, it is a distinction between pungent and tasteless, as a general theory, it may be an important lead compound in the development and research of future tasting ingredients There is.

(Effect of the fourth invention)
Since the inventor of the present application has been able to isolate the physiologically active substances of the first to third inventions from Myoga, naturally the production method of the fourth invention can be proposed as an effective production method of these physiologically active substances. .

(Effect of the fifth invention)
In the manufacturing method according to the fourth aspect of the present invention, it is particularly preferable that the extract is prepared by hexane extraction with myoga. However, it is a matter of course that the preparation of the extract by other various means can also be effectively employed.

(Effect of the sixth invention)
The physiologically active substances of the first to third inventions can be effectively produced also by a so-called crude enzyme method. As a preferred embodiment of such a crude enzyme method, the production method of the sixth invention can be exemplified.

  In addition, although this inventor is thinking about possibility that the bioactive substance of 1st invention-3rd invention can be effectively manufactured also by an organic synthesis method, until now, such an organic synthesis method However, the present invention has not yet completed a concretely effective embodiment.

(Effect of the seventh invention)
On the other hand, myogadiar, which is a well-known representative labdan-type diterpene aldehyde compound, exhibits a pungent taste, while the physiologically active substance of the third invention is tasteless, and both are similar in physiological activity. Therefore, considering the convenience of patients or consumers who dislike the pungent taste in the case of oral ingestion, the method of the sixth invention is the same as the seventh invention in that the labdane-type diterpene aldehyde compound having a pungent taste is physiologically active. It can be proposed as an invention of a method for tasting a pungent physiologically active substance which is said to be converted to a tasteless physiologically active substance while maintaining the above.

(Effect of the eighth invention)
Since the composition for health / medical use according to the eighth invention includes at least the physiologically active substance according to the first to third inventions, the pulverized ginger or the ginger extract as the active ingredient, the first to third inventions. The excellent health / medical effect of the physiologically active substance can be surely expected.

  As described above, since it has already been confirmed that the ginger contains gyoagial and diterpene aldehyde compounds in addition to the physiologically active substances according to the first to third inventions, the ginger pulverized product or the ginger extract In the composition for health / medical use, a synergistic effect by these compounds can be expected.

  As far as the inventor of the present application knows, a health care / medical composition containing a physiologically active substance according to the first to third inventions, or a plant pulverized product or an extract specifying Gyoza among ginger family plants has been heard. In that sense, the health / medical composition of the seventh invention, including a health / medical composition containing a crushed myoga or a ginger extract, is a health / medical product that can be expected to have a new health / medical effect. It is a medical composition.

  These health / medical compositions can be used at least for oral use, and can also be used for injection and other parenteral use. When it is made of pulverized myoga, attention may be required for parenteral use.

(Effect of the ninth invention)
As the ginger extract used in the eighth invention, a ginger hexane extract is particularly preferred as in the ninth invention.

(Effect of the tenth invention)
The health / medical composition according to the eighth or ninth invention can surely be expected to be effective as at least an antibacterial agent, an antithrombotic agent or an anti-inflammatory agent. In addition, the inventor of the present application estimates that it will be effective as an excellent renal function improving agent or anticancer agent from the points described in the “Effect of the first invention” column. Further, there is a possibility that new physiological activities of the physiologically active substances according to the first to third inventions will be revealed in the future, and there is a possibility that they can be used as health / medical agents for those uses.

(Effect of the 11th light)
Since the physiologically active substance according to the third invention is tasteless and exhibits a certain excellent physiological activity, the composition for health / medical use taken orally with the physiologically active substance according to the third invention as an active ingredient is Ideal for patients and consumers who dislike pungent taste.

  Next, modes for carrying out the first invention to the eleventh light of the present application will be described including the best mode. In the following, the term “present invention” simply refers to the corresponding invention group among the first invention to the eleventh light.

[Bioactive substances]
The physiologically active substance according to the present invention is isolated from Myoga, belongs to the category of labdan-type diterpene compounds, and is a major feature in that it is a trialdehyde compound as compared with the group of known labdan-type diterpene compounds. There is.

  The physiologically active substance isolated by the present inventor as such a labdan type diterpene trialdehyde compound is more specifically a 15,16,17-trial compound having a labdane skeleton structure, and more specifically, It is a compound shown in “Chemical Formula 2”. This compound, ie, (E) -8β (17) -labd-12-ene-15,16,17-trial [(E) -8β (17) -labd-12-ene-15,16,17-trial] Is called miogatrial.

  As described in the “Characterization” section of the examples, Myogatrial has been confirmed to have excellent antibacterial action, platelet aggregation inhibitory action, and human 5-lipoxygenase inhibitory action. As is well known, the platelet aggregation inhibitory action is directly linked to the antithrombotic action, and the human 5-lipoxygenase inhibitory action is directly linked to the anti-inflammatory action. Furthermore, it has been confirmed that myogatrial is tasteless, unlike myogadiar, which is a related compound exhibiting a known pungent taste.

  A variety of compounds can be specifically envisaged with respect to myogatrial, with slight changes and modifications in the chemical structure, while maintaining the characteristic point of the trialdehyde compound. For example, in Myogatrial, the atomic groups represented by position numbers 18, 19, and 20 in the “Chemical Formula 2” are all methyl groups, but one or more atomic groups are alkyl groups, alkenyl groups. Or the case where it is the alkyl group which has a hydroxyl group is assumed. It is considered that these compounds have a physiological activity equivalent to that of myogatrial and / or are likely to be tasteless.

  Furthermore, in the above-mentioned “Chemical Formula 2”, it is highly possible that various substituents in place of hydrogen can be introduced into carbon atoms such as position numbers 1 to 3, 6 to 7, and 9; Among these compounds, there is a possibility that a compound having a physiological activity equivalent to or higher than that of Myogatrial and / or tasteless and / or equivalent to or higher than Myogatrial in terms of stability or the like may be included. is there.

[Method for producing physiologically active substance]
The physiologically active substance according to the present invention can be produced by an extraction / purification method from Myoga or by enzymatic treatment of certain related compounds (precursor compounds). There is a possibility that it can be produced by an appropriate organic synthesis reaction.

  In the method for extracting and purifying from the ginger, the whole tissue of the ginger plant may be used as a raw material, but it is particularly preferable to selectively use the floret part or the underground stem part. As in the case of extraction from normal organisms, the raw material of myoga can be ground or pulverized in advance using a device such as a slicer, thereby increasing the efficiency of extraction of the target substance. Although an extraction solvent is not specifically limited, It is preferable to use an organic solvent, especially hexane. The extraction method or the type of the extraction device to be used can be arbitrarily selected, but it is preferable to use, for example, a homogenizer.

  Myoga trial cannot be separated only by the extraction operation from Myoga, and chromatographic operation or the like is required for the isolation and purification of Myoga Trial from the extract. In particular, there are very similar compounds, such as M. ginger, in M. ginger, which requires a very careful design of the isolation process.

  As will be described in detail in the “Examples” section, the present inventor isolated Myogatrial by combining silica gel flash chromatography (silica CC) and high performance liquid chromatography (HPLC) in a certain manner. Succeeded in doing.

  On the other hand, since the chemical structure of myogatrial was determined by the present inventors, it is also possible to synthesize myogatrial from certain related compounds.

  For example, in Myogatrial, an aldehyde group containing the carbon atom at position number 17 is bonded to the carbon atom at position number 8, whereas in Myogadiar, “formation” is described in the example section. The only difference is that the carbon atom at position number 8 and the carbon atom at position number 17 form an epoxy group, as shown as “3”. Therefore, by converting the epoxy group part to an aldehyde group by acting a crude enzyme extract of the myoga flower, a myotrial can be obtained. As a specific synthesis method therefor, the crude enzyme synthesis method according to the sixth invention can be exemplified.

  This crude enzyme synthesis method means that myogadiar, a physiologically active substance having a pungent taste, is converted to myogatrial, a tasteless physiologically active substance, and the pungent physiologically active substance is made tasteless while maintaining its physiological activity. This method also has important technical meaning.

  Next, for example, Galanal A and Galanal B shown as “Chemical Formula 4” in the Example section share the carbon atom at position number 8 in Myogatrial and the carbon atom at position number 15 constituting the aldehyde group. It is a structure closed by a bond. Therefore, in this case as well, for example, by applying a crude enzyme extract of Myoga florets to open this closed structure in Galanar A or Galanar B in a certain manner, Myoga Trial can be obtained. it can.

[Health / Medical Composition]
The health / medical composition according to the present invention contains the physiologically active substance according to any one of the first to third inventions as an active ingredient, or a crushed ginger as an active ingredient, or a ginger extract. Contains as an active ingredient. In this case, the distinction by the extraction solvent in the ginger extract is not limited, but an organic solvent extract, particularly a hexane extract is preferable.

  These health / medical compositions can be used as oral or parenteral pharmaceuticals for humans or non-human animals, as so-called supplements, or as health / health foods or their additives in a broader sense. As a medicine, use as an antibacterial agent, an antithrombotic agent or an anti-inflammatory agent is particularly preferable, but it may also be effective as a headache / antipyretic, allergy symptom relief agent, and the like. When used as a supplement or the like, there is a possibility that tastelessness of myoga triar will greatly appeal to consumers.

  In these health / medical compositions, the physiologically active substance according to any one of the first to third inventions, the crushed ginger or the ginger extract need only be contained as an “active ingredient”. Therefore, the health / medical composition can contain other optional components such as a bulking agent, an antioxidant and other stabilizers, a pH stabilizer in a liquid composition, a colorant, a preservative and the like. Further, the dosage form of the health / medical composition is not limited at all, and for example, powders, granules, capsules, tablets, liquids for internal use, ampoules for injection, and the like can be used.

  Next, examples of the present invention will be described. The technical scope of the present invention is not limited by the following examples.

[Example 1: Isolation of physiologically active substance from Myoga]
(Example 1-1: Pretreatment)
A fresh ginger floret was prepared, washed thoroughly with water and drained, and then sliced with a slicer to perform the next extraction efficiently, and used as a sample for extraction. Using the extraction sample 600 g thus prepared, the extraction operation was performed as follows.

(Example 1-2: Extraction)
After adding 3 times the amount of hexane to the above extraction sample and homogenizing, extraction was performed for 30 minutes while stirring on a magnetic stirrer at normal temperature and normal pressure. After the completion of this extraction process, the extraction residue was further extracted under the same conditions, and these extracts were mixed.

(Example 1-3: Isolation of physiologically active substance by chromatography)
The mixed extract was filtered and dehydrated, and concentrated to dryness using a rotary evaporator. This was dissolved in 2 ml of 15% acetone-hexane and loaded on a silica gel 7729 (MERCK) flash chromatography column 50 g equilibrated with hexane. The fraction containing Myogatrial was eluted with 300 ml of 15% acetone-hexane.

  The eluted fraction was concentrated to 1/10 with a rotary evaporator and purified by preparative HPLC (Shimadzu Corporation). The column is silica-60 (TOSOH) 7.8 × 300 mm, the mobile phase is 3.5% acetone-hexane, 4.0 ml / min. I went there. Detection is 232 nm and 30-34 min. The compound eluted in was recovered. Purification was repeated under the same conditions, and the recovered fraction was concentrated to dryness under a nitrogen stream to obtain an oily substance having a purity of 95 to 98%.

[Example 2: Determination of structure of physiologically active substance]
As a result of structural analysis of the crystal obtained in Example 1 by ¹³ C-NMR, this was a labdane-type diterpene trialdehyde compound represented by the above-mentioned “Chemical Formula 2”, and the atomic groups represented by position numbers 18, 19, and 20 thereof were respectively It was confirmed that it was (E) -8β (17) -labd-12-ene-15,16,17-trial which is a methyl group, and this was named miogatrial. Table ¹³ below shows the ¹³ C-NMR spectrum data.

[Example 3: Characterization of physiologically active substance]
Next, characterization of myogatrial was performed while comparing with other related compounds.

(Example 3-1: taste)
A small amount of a 95-98% pure solution of Myogatrial (ethanol solution with a concentration of 1 × 10 ⁻² mol) was spotted on a filter paper, air-dried, and then subjected to a sensory test with the tongue. As a result, pungent taste and other special tastes were not perceived and tasteless. Furthermore, the same sensory test was also performed for cases where the concentration of myogatrial was set to several concentrations exceeding 1 × 10 ⁻² mol, and it was confirmed that it was also tasteless.

  Incidentally, it has been confirmed that the above-mentioned related compound Myogadial (the structural formula is shown in “Chemical Formula 3” below) exhibits a pungent taste. On the other hand, it has been confirmed that other related compounds, galanal A and galanal B, are tasteless. The structural formula of Galanal A / B is shown in “Chemical Formula 4” below.

(Example 3-2: Antibacterial activity)
The antibacterial activity test with respect to the microorganisms shown in the following "Table 2" was done about Myouga trial, Myouga diar, Galanar A, and Galanar B. The antibacterial activity test is an agar dilution method, and the specific implementation conditions are as follows.

  1) Test sample: Several mg of each physiologically active substance (purity 95 to 98%) was accurately measured, dissolved in dimethyl sulfoxide (DMSO) to a concentration of 1 mg / ml, and filtered through a sterile filter. The filtrate was aseptically diluted several times with DMSO.

  2) Preparation of bacterial suspension and medium: Pre-culture at 37 ° C. for 13-15 hours in 3 ml of a neutral liquid medium (Nissui Pharmaceutical), and the turbidity at 660 nm was measured. A part thereof was diluted with physiological saline and used. Moreover, soybean casein digest (SCD agar medium, Daigo) with an agar concentration of 1.5% was prepared as an agar medium, and 20 ml was poured into a petri dish having a diameter of 9 cm to solidify.

  3) Antibacterial activity test: For 4 types of bacteria listed in Table 2, 100 μl of the pre-cultured bacterial suspension was added to 100 μl of the test sample of 1) above and top agar (0.6% NaCl − 0.7 ml of agar) was layered on 3 ml of the above SCD agar medium and cultured overnight at 37 ° C., and the appearance of colonies was observed with the naked eye to determine MIC. The minimum concentration that inhibits the growth of bacteria is referred to as the minimum inhibitory concentration (MIC). In principle, when even one colony was formed, it was considered that the bacteria had grown, and the lowest concentration (μg / ml) at which growth was completely inhibited was defined as MIC. The results of this antibacterial activity test are shown in Table 2.

In Table 2 above, the numerical value indicating “MIC (μg / ml)” as a unit is, for example, when the numerical value is “50”, the thinnest concentration of the test sample that completely inhibits the growth of bacteria is 50 μg / ml. It means that there is.

(Example 3-3: human 5-lipoxygenase inhibitory action)
Myogatrial and four other compounds were tested in vitro for human 5-lipoxygenase inhibitory activity associated with anti-inflammatory activity. The specific implementation method and implementation conditions of this test are as follows.

As an assay mixture, 0.1 mol Tris-HCl buffer (pH 8.0, 37 ° C.), 2 mmol CaCl ₂ , 2 mmol ATP, 10 μl sonicated phosphatidylcholine, 10 μl 5-lipoxygenase (protein), 10 μl measurement sample (DMSO Solution) was prepared and pre-incubated for 5 minutes at 30 ° C., and then 0.1 μmol of arachidonic acid was added. Immediately after the addition, incubation for 10 minutes was started at 30 ° C, and the reaction was stopped by adding 0.3 ml of cold methanol (-20 ° C) containing 0.2 nmol 13-hydroxylinoleic acid as an internal standard. .

  Then, 1 μl of 1N acetic acid was added, centrifuged (10,000 r.p.m., 10 minutes), and the supernatant was analyzed by reverse phase HPLC (Shimadzu Corporation). The column is CAPCELL PAK C18 (Shiseido) 4.6 × 150 mm, the mobile phase is methanol-water-acetic acid (80: 20: 0.01), and the flow rate is 0.8 ml / min. The detection was 232 nm. The amount of 5-hydroperoxyeicosatetraenoic acid (5-HPTEE) produced was quantified with Chromatopack (Shimadzu Corporation), and from the control [quercetin and NDGA (nordihydroguaiaretic acid)] based on the internal standard The inhibition rate was determined by comparison with the amount of 5-HPETE produced.

  The test results are shown in “Table 3” below. In Table 3, the calculation method of the inhibition rate indicated by “%” on the vertical axis is inhibition rate (%) = 100− (100 × measured sample and control added peak area value / measured sample and control non-added peak area value). .

Note that in Table 3 above, myoga triar is represented as “MT”. Moreover, the notation of “MD” indicates Myogadial. The notation “NDGA” indicates nordihydroguaiaretic acid known as an antioxidant. “Quercetin” is a polyphenolic compound known as vitamin P.

Example 3-4: Human platelet aggregation inhibitory action
Myogatrial, three other compounds (Myogadial, Galanar A, Galanar B) and two positive controls (aspirin and indomethacin: these are antipyretic / analgesic drugs marketed as is well known. ) Was tested in vitro for its inhibitory effect on human platelet aggregation associated with antithrombotic activity. The specific contents and conditions of this test are as follows.

  27 ml of healthy human venous blood was collected by a doctor. Immediately after adding 3 mL of sodium citrate solution (Citral for red sediment: manufactured by Yamanouchi Pharmaceutical), the mixture was centrifuged at 750 g for 10 minutes and platelet-rich plasma ( PRP) was prepared. The residue was centrifuged at 2000 g for 15 minutes to prepare platelet poor plasma (PPP). With a light scattering / turbidity combined aggregometer (PA-20, manufactured by Kowa Optimed), the turbidity difference between PRP and PPP is set to 100%, and ADP (adenosine diphosphate) is used as an aggregation initiator (initiator). Using the maximum aggregation rate (turbidity 2) of each sample added to the maximum aggregation rate (turbidity 1) of the control (PRP with no sample added), the human platelet aggregation inhibition rate is calculated as follows: did.

Human platelet aggregation inhibition rate (%) = 100− (turbidity 2 / turbidity 1) × 100
A curve at each concentration is prepared from the inhibition rate of human platelet aggregation, and the concentration at which human platelet aggregation is inhibited by 50% can be determined as an IC ₅₀ value (μM). It means that human platelet aggregation inhibitory activity is so strong that this value is small. Table 4 below shows miogatorial, miogadial, galanal A, galanal B, aspirin.
) And it shows an _{IC 50} value determined for indomethacin (indomethacin).

From Table 4, at least the following points can be pointed out. The inhibitory activities of Myogatrial and Myogadiar are almost the same. These have stronger inhibitory activity than aspirin, and are almost the same inhibitory activity as indomethacin. Inhibition activity is not recognized in galanar A and galanar B.

The present invention provides a novel labdane-type diterpene trialdehyde compound derived from Myouga. This compound is useful as an active ingredient such as an antibacterial agent, an antithrombotic agent or an anti-inflammatory agent, and has the advantage of being particularly suitable for oral administration because it is tasteless.

Claims (9)

Physiology active substances it is a lower following general Ru of compounds represented by the formula "Formula 1".
(In the above “Chemical Formula 1”, the atomic groups represented by position numbers 18, 19, and 20 are each a methyl group.) Gingae (Zingiber)
A method for producing a physiologically active substance, which comprises collecting the physiologically active substance according to claim 1 from an extract of mioga). The method for producing a physiologically active substance according to claim 2 , wherein the extract is prepared by hexane extraction for myoga. The bioactive substance according to claim 1 is produced by allowing a crude enzyme extract prepared from a myoga flower bud to act on a raw material liquid containing a labdane-type diterpene aldehyde compound having an epoxy group bonded to an aromatic ring, A method for producing a physiologically active substance, which is collected. A labdane-type diterpene aldehyde compound having the above-mentioned pungent taste is prepared by allowing a crude enzyme extract prepared from a myoga flower bud to act on a raw material solution containing a pungent-type labdane-type diterpene aldehyde compound having an epoxy group bonded to an aromatic ring. , while maintaining its physiological activity, tasteless method of pungent physiologically active substance, characterized in that to convert the raw physiologically active substance according to claim 1. A health / medical composition comprising, as an active ingredient, at least the physiologically active substance according to claim 1 , a crushed ginger or a ginger extract. 7. The health / medical composition according to claim 6 , wherein the ginger extract is a ginger hexane extract. The health / medical composition according to claim 6 or 7 , wherein the health / medical composition is an antibacterial agent, an antithrombotic agent or an anti-inflammatory agent. A health / medical composition comprising the physiologically active substance according to claim 1 as an active ingredient and ingested orally.