JP2013241473A - Adrenomedullin production enhancing agent - Google Patents

Abstract

PROBLEM TO BE SOLVED: To find substances having the capability to effectively enhance the production of adrenomedullin, and to provide an adrenomedullin production enhancing agent using the same.SOLUTION: An adrenomedullin production enhancing agent contains, as effective ingredients, ginsenoside, sanshool and/or shogaol.

Description

  The present invention relates to an adrenomedullin production enhancer, and more specifically, adrenomedullin effective in the prevention and treatment of diseases such as Crohn's disease by promoting the production of adrenomedullin, an intestinal peptide having an effect of increasing blood flow and an anti-inflammatory effect. The present invention relates to a production enhancer.

  Adrenomedullin was discovered in 1993 as a circulating regulatory peptide having a strong vasodilatory action (Patent Document 1). Adrenomedullin is produced from various organs such as the circulatory system and digestive system, and has been reported to have important physiological activities such as vasodilation, angiogenesis, antibacterial action, anti-enteritis, gastric mucosa protection, and thrombus formation inhibition. Yes. Therefore, the effect of adrenomedullin administration in the treatment of various diseases has been confirmed, and is effective for, for example, myocardial disorders, non-bacterial inflammatory diseases, pulmonary hypertension, bone disorders, myometrial contractions, dysuria, etc. It has been reported (patent documents 2 to 7).

  However, since adrenomedullin is a peptide, its production cost is high to formulate it, and when adrenomedullin is administered directly, it becomes a dosage form of injection or infusion, so it is strictly free from contamination with endotoxins and other foreign substances. Manufacturing technology and management are required. Adrenomedullin has a short half-life in the blood of several tens of minutes and is expected to be administered in a considerable amount if it is expected to produce a therapeutic effect, but administration of a large amount of intravenous adrenomedullin has an acute antihypertensive effect. There is a risk of triggering. Furthermore, when expecting adrenomedullin exposure to specific organs, there were various problems such as requiring targeting technology. Therefore, a highly safe drug that can be produced at low cost and can enhance an adrenomedullin production system that is constitutively operating in vivo by oral administration has been desired. The drug has not been known so far.

Japanese Patent No. 2774769 International Publication No. 00/078338 Pamphlet International Publication No. 00/0783339 Pamphlet Special Table 2002-540216 Japanese Patent Laid-Open No. 2003-300899 JP 2006-290777 A JP 2006-290814 A

  Accordingly, an object of the present invention is to find a substance that can be administered orally and effectively enhances the production of adrenomedullin in vivo, and to provide an adrenomedullin production enhancer using this substance.

  In order to solve the above-mentioned problems, the present inventors have eagerly searched for a substance having an effect of enhancing adrenomedullin production. Has been found, and the present invention has been completed.

That is, the present invention
Compound represented by the following formula (1)
(Shown wherein ^R 1, ^{R 2} and ^{R 3} are the same or different and each represents a hydrogen atom, a hydroxyl group, -O-Glc, -O-Glc -Glc, the -O-Glc-Ara, or -O-Glc-Rha. (However, Glc represents a glucose residue, Ara represents an arabinose residue, and Rha represents a rhamnose residue.)
And a compound represented by the following formula (2) and / or a compound represented by formula (3)
(In the formula, R ⁴ represents a hydrogen atom or a hydroxyl group, m is 1 or 2, and a wavy line indicates a Z-configuration or an E-configuration)
(Wherein n is 4, 6 or 8)
Is an adrenomedullin production enhancer.

  The adrenomedullin production enhancer of the present invention can effectively promote the production of adrenomedullin having various physiological activities such as blood flow increasing action and anti-inflammatory action. Therefore, the adrenomedullin production enhancer of the present invention can be used for the treatment of various diseases. For example, the action of increasing blood flow in the intestinal tract effectively prevents Crohn's disease, which is an ischemic digestive disease. Can be treated. The adrenomedullin production enhancer of the present invention is derived from a natural product and can be administered orally, so it has high safety and excellent effect persistence, and particularly has a high effect of enhancing adrenomedullin production in the digestive tract.

In Example 1, it is a figure which shows the measurement result of the adrenomedullin density | concentration in the plasma of portal vein blood. Note that * means a risk rate of 5% or less, and ** means a risk rate of 1% or less. In Example 2, it is a figure which shows the increase rate of VC by Daikenchuto administration. In Example 2, it is a figure which shows the increase rate of VC by ginseng administration. In Example 2, it is a figure which shows the increase rate of VC by psoriasis administration. In Example 2, it is a figure which shows the increase rate of VC by yam administration. In Example 2, it is a figure which shows the increase rate of VC by psoriasis and ginseng administration. In Example 2, it is a figure which shows the increase rate of VC by carrot and yam administration. In Example 2, it is a figure which shows the increase rate of VC by psoriasis and yam administration. In Example 3, it is a figure which shows the increase rate of VC of an ADM antagonist pretreatment group and the Daikenchuto administration group. In Example 4, it is a figure which shows the increase rate of VC by hydroxysanshool single administration, hydroxysanshool, and a ginseng extract combined administration. In Example 4, it is a figure which shows the increase rate of VC by ginsenoside Rb1 single administration, hydroxysanshool, and ginsenoside Rb1 combined administration. In Example 4, it is a figure which shows the increase rate of VC by ginsenoside Rg1 single administration, hydroxysanshool, and ginsenoside Rg1 combined administration. In Example 5, it is a figure which shows the increase rate of VC by 6-shogaol single administration, hydroxysanshool, and 6-shogaol combined administration. In Example 6, it is a figure which shows the measurement result of a necrosis area. In Example 6, it is a figure which shows the measurement result of adhesion frequency. In Example 6, it is a photograph of the large intestine 3 days after intestinal inflammation induction. In Example 7, it is a figure which shows the measurement result of liver fibrosis. In Example 7, it is a figure which shows the fixed_quantity | quantitative_assay result of the amount of hyaluronic acid in blood. In Example 7, it is a figure which shows the fixed_quantity | quantitative_assay result of the amount of hydroxyproline in a liver.

  As one of the active ingredients used in the adrenomedullin production enhancer of the present invention, a compound represented by the following general formula (1) (hereinafter sometimes referred to as “compound (1)”) is used.

(Wherein R ¹ , R ² and R ³ are the same or different and each represents a hydrogen atom, a hydroxyl group, —O—Glc, —O—Glc-Glc, —O—Glc-Ara or —O-Glc-Rha). (However, Glc represents a glucose residue, Ara represents an arabinose residue, and Rha represents a rhamnose residue.)

Specifically, as the compound (1), R ¹ and R ² are —O—Glc-Glc, R ³ is a hydrogen atom, ginsenoside Rb ₁ , R ¹ is —O—Glc-Glc, and R ² is — O-Glc-Ara, ginsenoside Rb ₂ or Rc in which R ³ is a hydrogen atom, R ¹ is —O-Glc-Glc, R ² is —O-Glc, and ginsenoside Rd or R ^{1 in} which R ³ is a hydrogen atom Ginsenoside Rg ₁ in which R ² is —O—Glc, R ³ is —O—Glc-Rha, Ginsenoside Re, R ¹ is hydroxyl, R ² is —O—Glc, and R ³ is —O—Glc, R ¹ and ^{R 2} are hydroxyl group and a ginsenoside such ginsenoside _Rg 2, ^{R 1} and ^{R 2 R 3} is -O-Glc-Rha is hydroxyl, ^{R 3} and ginsenoside Rh ₁ is -O-Glc . Of these, ginsenoside Rb ₁ and ginsenoside Rg ₁ are preferred.

  These compounds can be isolated by known methods (reference: Japanese Oriental Medicine Journal Vol. 35, No. 1 Page. 1-22 (1984), etc.), or commercially available products can be used.

The adrenomedullin production enhancer of the present invention includes a compound represented by the above formula (1), a compound represented by the following formula (2) (hereinafter sometimes referred to as “compound (2)”) and / or A compound represented by the formula (3) (hereinafter sometimes referred to as “compound (3)”) is used in combination.
(In the formula, R ⁴ represents a hydrogen atom or a hydroxyl group, m is 1 or 2, and a wavy line indicates a Z-configuration or an E-configuration)
(Wherein n is 4, 6 or 8)

Specifically, as the compound (2), R ⁴ is a hydrogen atom, m = 1, α-sanshool having a Z-configuration in the wavy line portion, R ⁴ is a hydrogen atom, m = 1, and the wavy line portion is E−. Β-sanshool in configuration, R ⁴ is a hydrogen atom, m = 2, w-line portion is γ-sanshool in Z-configuration, R ⁴ is hydroxyl group, m = 1, w-line portion is in Z-configuration hydroxy- α-sanshool, R ⁴ is hydroxyl group, m = 1, wavy portion is hydroxy-β-sanshool, R ⁴ is hydroxyl group, m = 2, wavy portion is Z-configuration hydroxy-γ- Sanshools such as Sanshool are listed. Of these, hydroxy-α-sanshool and hydroxy-β-sanshool are preferably used.

  These compounds are prepared by known methods (Literature: Biosci Biotechnol Biochem: Vol. 69 No. 10 Page. 1951-1957 (2005), Biological & Pharmaceutical Bulletin (2007), 30 (1), 205-207, Phytotherm 97). 44 (6), 1125-1127, etc.).

  On the other hand, specific examples of compound (3) include 6-shogaol, 8-shogaol, 10-shogaol and the like. Of these, 6-shogaol is preferred.

  These compounds are isolated or synthesized by a known method (literature: Bulletin of the Chemical Society of Japan (1976), 49 (5), 1453-1454, Japanese Patent Application No. Sho 63-137743, etc.), or commercially available products are obtained. Can be used.

  The adrenomedullin production enhancer of the present invention comprises the compound represented by the above formula (1) and the compound represented by the formula (2) and / or the compound represented by the formula (3) as active ingredients. It can be produced by appropriately mixing with a pharmaceutical carrier and formulating it as an oral or parenteral preparation.

  Oral preparations can be powders, powders, granules, tablets, capsules, soft capsules, liquids, etc., and pharmaceutical carriers corresponding thereto, such as starch, lactose, sucrose, mannitol, carboxymethylcellulose Corn starch, inorganic salt, etc. can be used. In preparation of the oral preparation, a binder, a disintegrant, a surfactant, a lubricant, a fluidity promoter, a corrigent, a colorant, a fragrance and the like can be further blended.

  Parenteral preparations are also produced according to conventional methods. As diluents, distilled water for injection, physiological saline, aqueous glucose solution, vegetable oil for injection, sesame oil, peanut oil, soybean oil, corn oil, propylene glycol, polyethylene glycol and the like are generally used. Can be used. Furthermore, bactericides, preservatives, stabilizers and the like can be added as necessary.

  In the adrenomedullin production enhancer of the present invention, the compounding amount of the compound (1) as an active ingredient varies depending on the type of compound, target disease, disease level, patient age, etc., for example, when ginsenoside Rb1 is used, The daily dose per adult is about 1 mg to 1 g. The compounding amount when using the compound (2) as a component used in combination with the compound (1) is about 1 mg to 1 g as a daily dose per adult, and the compounding amount when using the compound (3) as a combination component. Is about 1 mg to 1 g. On the other hand, the compounding amount when the compound (2) and the compound (3) are used in combination is about 2 mg to 2 g in total.

  Moreover, the adrenomedullin production enhancer of this invention can also use the crude drug containing the said compound (1), the crude drug containing a compound (2), and / or the crude drug containing a compound (3) as an active ingredient. Examples of herbal medicines containing the compound (1) include carrots. The herbal medicine containing the compound (2) includes yam, and the herbal medicine containing the compound (3) includes psoriasis. These carrots, yam, and dried rice cake are all known as herbal medicine ingredients, and commercially available products can also be used.

  Preparation of the adrenomedullin production enhancer of the present invention was obtained by chopping or crushing ginseng and yam and / or dried rice cake and then uniformly mixing them, or further extracting with an appropriate solvent as necessary. The extract is dried and mixed with koji as needed.

  The amount of ginseng in the adrenomedullin production enhancer of the present invention is about 1 mg to 1 g as a daily dose per adult. Moreover, when using yam as a component to be used in combination with carrot, the blending amount is about 1 mg to 1 g, and when using ginger, it is about 1 mg to 1 g. On the other hand, the compounding amount when using yam and dried rice cake together is about 2 mg to 2 g in total.

  As a Kampo prescription containing ginseng and yam and / or dried rice cake, Daikenchuyu, Tokiyu, Hanatsushinshinyu, ginsengyu, Hankaba Shirasagi Tenmayu, Hochuekkito, Keishijinjinto, There are Daifufuto, Hourento, etc., which can also be used as an active ingredient of the adrenomedullin production enhancer of the present invention. An example of the general structure of Daikenchuyu is a dry extract of mixed herbal medicine (Daikenchuto extract powder) in a ratio of 15 g of Daikenchuyu: dried ginger: carrot: yam = 5: 3: 2. Contains 25 g and 10 g of glue.

  The adrenomedullin production-enhancing agent of the present invention is produced by using the above ginseng and dried ginger and / or yam as active ingredients, mixing with other pharmaceutical carriers as appropriate, and formulating this as an oral or parenteral preparation. can do. As the pharmaceutical carrier, the same components as those described above can be used.

  By administering the adrenomedullin production enhancer of the present invention obtained as described above, adrenomedullin production in various organs / tissues can be promoted. Adrenomedullin has various physiological activities such as vasodilation, angiogenesis, antibacterial, anti-enteritis, gastric mucosal protection, thrombus formation suppression, etc., through such actions, such as ulcerative colitis and Crohn's disease It is effective for the prevention and treatment of diseases such as inflammatory bowel disease, dysuria, myometrial contraction, bone disorder, hypertension, myocardial disorder, non-bacterial inflammatory disease and hepatitis.

  EXAMPLES Next, although an Example is given and this invention is demonstrated in more detail, this invention is not restrict | limited at all by these Examples. In addition, the extract powder used in the Examples was obtained by extracting water from a herbal medicine or a mixture of herbal medicines according to a conventional method.

Example 1
Daikenchuto enhances adrenomedullin production:
SD male rats (8-10 weeks old, body weight 300-400 g) were used (n = 16). An aqueous glue solution was prepared by adding glue to distilled water to a concentration of 480 mg / mL. Daikenchuto extract powder was weighed and added to the aqueous koji solution at a concentration of 60 mg / mL at the time of use, and uniformly dispersed by stirring at room temperature for 30 minutes to prepare a test sample. A 5 mL / kg test sample kept at 37 ° C. was administered into the duodenum via a cannula, and portal blood was collected 0, 15, 30, 60, and 90 minutes after administration. Distilled water was administered to the control group in the same manner. About 5 mL of portal vein blood was collected in a 15 mL centrifuge tube made of polypropylene (PP) into which 100 μL of edetic acid (EDTA) / aprotinin solution (containing 5 mg EDTA-2Na and 2500 KIU aprotinin) was dispensed. Centrifugation was performed at 4 ° C. for 15 min. To a 5 mL centrifuge tube made of PP for high speed centrifuge, 2 mL plasma and 0.16 mL amount of an acidifying solution having the following composition were added to acidify the plasma sample.

  The supernatant centrifuged at 7000 g, 4 ° C., and 20 min was passed through an activated Sep-Pak cartridge column (C-18 column, Waters WAT020805), and adrenomedullin (ADM) was adsorbed onto the column. After washing twice with 2.5 mL of column wash (0.1% trifluoroacetic acid (TFA) aqueous solution), the adsorbed ADM was eluted with 2 mL of column eluent (0.1% TFA methanol solution). It was. The eluate was received in a 5 mL centrifuge tube, loaded into a centrifuge under reduced pressure, evaporated to dryness, and stored at −80 ° C. until EIA (Enzyme immunoassay) measurement. In quantification of ADM, it measured using RatADM EIA kit (EK-010-08 by Phoenix Pharmaceuticals). The evaporated and dried sample was dissolved in a 400 uL amount of EIA buffer, then subjected to heat treatment at 90 ° C. for 15 min, and centrifuged at 2000 G for 20 min to obtain an EIA sample. Data were expressed as mean ± standard error (SEM). Data were subjected to Two-way ANOVA, followed by Dunnett's multiple comparison or student t test. A significance level of 5% or less was adopted as the significance level. Changes in plasma ADM concentration are shown in Table 1 and FIG.

(Acidified solution composition)
NaCl 100 mg (final concentration: 1%)
Trifluoroacetic acid (TFA) 0.1 mL (final concentration: 1%)
Formic acid 0.5 mL (final concentration: 5%)
8.0 mL of 1N hydrochloric acid (final concentration: 80%)
Distilled water 1.4 ml
10 mL total

  The portal blood plasma ADM concentration collected immediately before enema of the test sample was 30.4 ± 2.7 pg / mL, while 15, 30, 60, 90 min after administration of the test sample Daikenchuto. The later collected groups showed significant increases over time at 43.3 ± 3.0, 54.3 ± 3.7, 53.0 ± 4.4, and 44.1 ± 3.3 pg / mL, respectively. In comparison between the groups of distilled water administration and Daikenchuto administration, a significant difference (p <0.01) was recognized at 15, 30, and 60 min after administration.

Example 2
Blood flow increasing effect by Daikenchuto and its constituent crude drugs:
SD male rats (9-11 weeks old, body weight 260-350 g) were used (n = 3 or 6). The test sample (Daikenchuto) was prepared by suspending the paste and Daikenchuto extract powder in distilled water to a concentration of 160 mg / mL of glue and 20 mg / mL of Daikenchuto extract powder. In addition, for ginseng extract powder, dried ginger extract powder, and yam extract powder, test samples of each herbal medicine alone in distilled water, ginseng extract powder, dried ginger extract powder, and yam extract powder respectively at concentrations of 6 mg / mL, 10 mg / It was prepared by adding to 4 mL / mL. Furthermore, the test sample which used two crude drugs together among the above-mentioned three crude drugs, ginseng extract powder 6mg / mL and dried ginger extract powder 10mg / mL, ginseng extract powder 6mg / mL and yam extract powder 4mg / mL in distilled water, It was prepared by adding to a concentration of 4 mg / mL of yam extract powder and 10 mg / mL of dried salmon extract powder. 5 mL / kg of a test sample kept at 37 ° C. was inserted through the cecum in advance and administered via the 18 G Surfflow indwelling needle in the colon, 0.5 to 1 hour after the operation when the blood flow base value was stabilized. did.

  A polyethylene tube having an inner diameter of 0.58 mm was inserted from the left common carotid artery of the anesthetized rat, and the tip was placed in the left ventricle. After placing a tracheal cannula in the trachea, it was connected to a respirator (SN-480-7 manufactured by Shinano Manufacturing Co., Ltd.) and artificial respiration was performed at a frequency of 60 RPM. The body temperature of the rat was maintained at 37 ± 0.5 ° C. with a temperature controller (NS-TC10, manufactured by Neuroscience). An incision was made along the abdominal midline of the rat and the cecum was pulled out of the body, and then the distal large intestine was exposed. The lower part of the large intestine was lightly lifted with tweezers, and in a state where stool was present in the large intestine, 4 sites were tied to the ends of the left and right abdominal rectus muscles cut with 5-0 sutures. The blood flow probe was set on the upper part of the distal large intestine where the blood flow probe was fixed, and the blood flow (flow, mass, velocity) was measured using a laser tissue blood flow meter (ALF21N manufactured by Advance Co., Ltd.). To prevent dryness, the entire abdomen including the blood flow probe was covered with a wrap. In addition, blood pressure and heart rate are monitored simultaneously with blood flow, and each measured value is measured by a patient monitoring device (BP-508 manufactured by COLIN) [biological amplifier (Nihon Kohden)], a data recording device (Power Lab / 800 manufactured by AD instruments). [8/30]), and recorded in data analysis software (Char v3.6 [v5.4.2] manufactured by AD instruments). Blood flow measurement was performed up to 90 minutes after drug administration, and the rate of increase (%) was calculated using basal conductance (VC) obtained by dividing flow by blood pressure as an index of blood flow. The increase rate of VC by each test sample is shown in Table 2 and FIGS.

  The Daikenchutou administration group showed an increase in VC 15 to 90 minutes after administration. In the group of Daikenchuto's constituent crude drugs alone, ginseng showed a transient increase in blood flow 15 minutes after administration, and yam showed a weak increase effect peaking at 75 minutes after administration. Psoriasis did not show a clear increase in blood flow. Regarding the combination of herbal medicines, the combination of ginseng and psoriasis and the combination of ginseng and yam showed a significantly higher VC increase than each single administration. No change in the rate of increase was observed with the combination of psoriasis and yam.

Example 3
Effect of ADM antagonist pretreatment on the blood flow increasing effect of Daikenchuto:
SD male rats were divided into two groups: a group with Daikenchuto and an ADM antagonist pretreatment group (n = 7 to 8). Using distilled water, prepare a gelatin solution with a concentration of 160 mg / mL, and weigh Daikenchuto extract powder to a concentration of 20 mg / mL, and stir at room temperature for at least 30 min in the same solution. And then uniformly dispersed to prepare Daikenchuto (TJ-100). This was administered to the Daikenchuto group in the colon at an amount of 900 mg / 5 mL / kg. On the other hand, in the ADM antagonist pretreatment group, a human ADM antagonist (h.ADM22-52, Peptide 4302-v) was dissolved at 30 μmol / L in physiological saline, and 30 nmol / 1 mL from a cannula attached under anesthesia. After 15 minutes, Daikenchuto was administered intracolonically at 900 mg / 5 mL / kg.

  A polyethylene tube with an inner diameter of 0.8 mm was inserted from the left common carotid artery of the anesthetized rat, and the tip was placed in the left ventricle. After placing a tracheal cannula in the trachea, it was connected to a respirator (SN-480-7) and artificial respiration was performed at a frequency of 60 RPM. The body temperature of the rat was maintained at 37 ± 0.5 ° C. with a temperature controller (NS-TC10). Thereafter, the VC increase rate (%) was calculated in the same manner as in Example 2. The increase rate of VC in each group is shown in Table 3 and FIG. The measurement results were expressed as an average value ± standard error (SEM). The effect comparison was tested by one-way analysis of variance (ANOVA) followed by Scheffe all-group comparison. A significance level of 5% or less was adopted as the significance level.

  In the ADM antagonist treatment, a significant VC reduction of 30.8 to 99.5% was shown in the Daikenchuto alone administration group from 30 to 90 minutes after administration. Thus, since the blood flow increasing action of Daikenchuto disappeared by the ADM antagonist pretreatment, it was shown that ADM contributes to the blood flow increasing action.

Example 4
Blood flow increasing action of ingredients in herbal medicine (1):
SD male rats (8-10 weeks old, body weight 240-360 g) were used (n = 2-6). Hydroxy-β-sanshool, ginsenoside Rb1 or ginsenoside Rg1 was added to 1% Tween 80 aqueous solution so as to have concentrations of 0.06 mg / mL, 0.2 mg / mL and 0.2 mg / mL, respectively. Test samples for administration. Moreover, about the test sample of combined administration of hydroxysanshool and ginseng extract, ginsenoside Rb1 or ginsenoside Rg1, hydroxysanshool having a concentration of 0.06 mg / mL, ginseng extract powder, ginsenoside Rb1 or 1% Tween80 aqueous solution Ginsenoside Rg1 was prepared by adding to concentrations of 6 mg / mL, 0.2 mg / mL and 0.2 mg / mL, respectively. 5 mL / kg of a test sample kept at 37 ° C. was inserted through the cecum in advance and administered via the 18 G Surfflow indwelling needle in the colon, 0.5 to 1 hour after the operation when the blood flow base value was stabilized. did. The control group received 1% Tween 80 aqueous solution.

  A polyethylene tube with an inner diameter of 0.8 mm was inserted from the left common carotid artery of the anesthetized rat, and the tip was placed in the left ventricle. After placing a tracheal cannula in the trachea, it was connected to a respirator (SN-480-7) and artificial respiration was performed at a frequency of 60 RPM. The body temperature of the rat was maintained at 37 ± 0.5 ° C. with a temperature controller (NS-TC10). Thereafter, the VC increase rate (%) was calculated in the same manner as in Example 2. The increase rate of VC by each test sample is shown in Table 4 and FIGS. The measurement results were expressed as an average value ± standard error (SEM). The effect comparison was tested by one-way analysis of variance (ANOVA) followed by Scheffe all-group comparison. A significance level of 5% or less was adopted as the significance level.

  Hydroxysanshool did not show an increase in blood flow when administered alone, but when used in combination with ginseng extract, a significant increase in VC of 53.7 to 109.0% was observed with respect to the control group from 30 to 90 minutes after administration. Indicated. Moreover, compared with the hydroxysanshool single administration group, the significant VC increase of 45.4, 63.4, and 46.8% was shown in 45, 60, and 90 minutes after administration, respectively. The peak at the time of combination was 75 minutes after administration, which was consistent with the blood flow increase pattern observed with Daikenchuto or Yamatake extract alone in Example 1. In addition, the combined administration group of hydroxysanshool and ginsenoside Rb1 or Rg1 also showed a significant increase in VC compared to the hydroxysanshool single administration group. From this result, it was clarified that at least hydroxysanshool was involved as an active ingredient of yam, and it was speculated that ginseng plays an important role in the expression of the activity.

Example 5
Blood flow increasing action of ingredients in herbal medicine (2):
SD male rats (8-10 weeks old) were used. A test sample was prepared by adding 6-shogaol to a 1% Tween 80 aqueous solution to a concentration of 0.4 mg / mL. In addition, a test sample was prepared by adding ginsenoside Rb1 and 6-shogaol to 1% Tween 80 aqueous solution so as to have concentrations of 0.2 mg / mL and 0.4 mg / mL, respectively. 5 mL / kg of a test sample kept at 37 ° C. was inserted through the cecum in advance and administered via the 18 G Surfflow indwelling needle in the colon, 0.5 to 1 hour after the operation when the blood flow base value was stabilized. did. The control group received 1% Tween 80 aqueous solution.

  A polyethylene tube with an inner diameter of 0.8 mm was inserted from the left common carotid artery of the anesthetized rat, and the tip was placed in the left ventricle. After placing a tracheal cannula in the trachea, it was connected to a respirator (SN-480-7) and artificial respiration was performed at a frequency of 60 RPM. The body temperature of the rat was maintained at 37 ± 0.5 ° C. with a temperature controller (NS-TC10). Thereafter, the VC increase rate (%) was calculated in the same manner as in Example 2. The increase rate of VC by each test sample is shown in Table 5 and FIG.

  When 6-shogaol was administered alone, it showed an increase in blood flow that peaked at 30 minutes immediately after administration, and in combination with ginsenoside Rb1, an increase in blood flow peaked at 45 minutes was observed.

Example 6
Anti-enteritis action in an animal model of Crohn's disease:
BALB / c male mice (20-25 g) were used (n = 6 or 7). A test sample (Daikenchuto) was prepared by adding gelatin and Daikenchuto extract powder to distilled water to a concentration of 80 mg / mL and 10 mg / mL, respectively.

  2,4,6-trinitro-benzene sulfonic acid (TNBS, manufactured by Tokyo Chemical Industry Co., Ltd.) was weighed into a PP tube, added with 50% aqueous EtOH solution, and stirred and dissolved at a concentration of 15 mg / mL. . A 1 mL Terumo syringe was connected to a feeding tube (SF-FT0380FG, Fr3.5, outer diameter 1.2 mm, manufactured by Terumo Corporation). A 3.5 cm position from the tip of the cannula was marked with an oily magic. During 15-30 min, mice were left in a small net cage to induce defecation, and anesthesia was performed by intraperitoneal administration of 55 mg / kg pentobarbital and 0.75 mg / kg atropine (manufactured by Sigma). After leaving for 15-30 min, a TNBS-injected cannula with olive oil attached to the tip (with olive oil attached to the tip) was slowly inserted to a depth of 3.5 cm. While the cannula was inserted with the office clip, the anus was stopped and the intestinal lumen was sealed. The mouse was suspended, and 1.5 mg / 0.1 mL / head of TNBS was slowly injected (2 sec / 0.1 mL), and maintained for 30 sec to induce enteritis.

900 ml / kg Daikenchuto was forcibly orally administered at 8, 24, 32, 48, and 56 hours after TNBS enema. Distilled water was similarly administered to the control group. On day 3 (72 hours later) from the onset of enteritis due to TNBS enema, laparotomy was performed and the presence or absence of adhesion was observed. The anus was collected from the cecum and a photograph was taken (digital camera D100 manufactured by Nikon Corporation). Cut along the longitudinal muscle, the contents were washed away with physiological saline, and the photo was taken again with the mucosa facing up. Photographic images were captured with image analysis software (Image J) and the necrotic area (cm ² ) on the luminal surface was measured. In addition, the pathological condition was similarly evaluated about the untreated group which does not perform enteritis induction processing and test sample administration.

  Necrotic area measurement data were evaluated for differences between groups using Welch's t-test. For the significance between groups, a risk rate of 5% or less was adopted as a significance level. The measurement results of the necrotic area are shown in FIG. 14, and the adhesion frequency is shown in FIG. Moreover, the photograph of the large intestine 72 hours after TNBS enema is shown in FIG.

  In the control group, in the macroscopic evaluation of the large intestine 3 days after the onset of enteritis, adhesion formation with the mesentery of the large intestine and a severe necrosis image were observed on the luminal mucosa side. On the other hand, in the Daikenchutou administration group, the frequency of adhesion of the large intestine was remarkably reduced and the severe necrotic area inside the lumen was significantly reduced by oral administration 5 times after 8 hours of TNBS treatment.

Example 7
Effects of Daikenchuto in a rat cirrhosis model
A 300 mg / L concentration of thioacetamide (TA) solution was administered to SD male rats for 20 weeks to produce TA-induced cirrhosis. Daikenchuto extract powder was given as a 50 or 200 mg / kg / day dose of mixed diet from the beginning of TA treatment to the end of the test from the 10th week. For pathological evaluation, blood and liver tissues were collected at 10 and 20 weeks after TA treatment. The collected liver tissue was stained with a hematoxylin-eosin (HE) staining method and a sirius red staining method, a pathological image was taken into image analysis software (Image J), and liver fibrosis was evaluated by area ratio. The amount of hyaluronic acid in the blood was quantified using a hyaluronic acid ELISA kit (manufactured by Cosmo Bio Inc.). In addition, the amount of hydroxyproline in the liver was determined by the Norman Logan method.

  All test results were expressed as mean ± standard deviation (S.D.). Statistical analysis was performed by Dunnett's multiple comparison between the groups that had been drinking TA solution for 20 weeks, and a significance level of 5% or less was evaluated as a significant level.

  The evaluation results of liver fibrosis are shown in FIG. 17, the measurement results of the amount of blood hyaluronic acid are shown in FIG. 18, and the quantification results of hydroxyproline in the liver are shown in FIG.

  In the hepatitis control group at 10 and 20 weeks after TA treatment, hepatic fibrosis markers, blood hyaluronic acid and hepatic hydroxyproline, increased significantly compared to the untreated group, respectively, and the 20 week group was particularly marked . In the hepatitis control group at 20 weeks after TA treatment, sirius red-positive histopathology was prominent, indicating findings peculiar to cirrhosis. On the other hand, in the group in which Daikenchuto was administered from the 10th week of TA treatment and evaluated at the 20th week, the increase in blood hyaluronic acid and hepatic hydroxyproline was significantly reduced compared with the control group. It exhibited a clear medicinal effect. The same effect was confirmed by HE and Sirius red staining. Therefore, Daikenchuto exhibited a profound medicinal effect by inhibiting progressive liver fibrosis in a TA-induced cirrhosis model.

Product example 1
50 g of ginsenoside Rb1 and 50 g of hydroxy sanshool were mixed with 270 g of lactose, 120 g of microcrystalline cellulose, and 10 g of magnesium stearate, and this mixture was tableted with a single tableting machine to produce a tablet having a diameter of 9 mm and a weight of 250 mg. .

  Each tablet contains 25 mg of ginsenoside Rb1 and hydroxysanshool, and is taken 3 to 10 tablets a day according to symptoms.

Product example 2
25 g of ginsenoside Rg1 and 25 g of hydroxy sanshool were mixed with 950 g of corn starch, kneaded with water, granulated on a screen having a 1 mm × 1 mm network, and dried to give granules.

  1 g of this granule contains 25 mg each of ginsenoside Rg1 and hydroxysanshool, and is taken 2 to 6 g per day according to symptoms.

Product example 3
50 g of shogaol and 50 g of hydroxysanshool were mixed with 210 g of lactose, 120 g of starch, 50 g of talc and 20 g of magnesium stearate, and 250 mg each was filled into a hard capsule to produce a capsule.

  One capsule contains 25 mg each of shogaol and hydroxysanshool, and is taken 3 to 10 capsules a day according to symptoms.

Product example 4
50 g of ginseng extract and 50 g of yam extract were mixed with 270 g of lactose, 120 g of microcrystalline cellulose, and 10 g of magnesium stearate, and the mixture was tableted with a single tableting machine to produce tablets with a diameter of 9 mm and a weight of 250 mg.

  Each tablet contains 25 mg of ginseng extract and yam extract, and is taken 3 to 10 tablets a day according to the symptoms.

Product example 5
Ginseng extract, 25 g of yam extract and 25 g of dried koji extract were mixed with 925 g of corn starch, kneaded with water, granulated on a screen having a 1 mm × 1 mm mesh, and dried to give granules.

  1 g of this granule contains 25 mg each of ginseng extract, yam extract, and dried koji extract, and is taken in divided doses of 2 to 6 g per day according to symptoms.

Product example 6
Production of Daikenchuto extract granules:
Cut herbal medicine, weigh and prepare 3 kg of carrot, 2 kg of yam, and 5 kg of dried salmon, add to about 12 times the amount of purified water, raise the temperature to 95-100 ° C. with stirring, and then raise the temperature for about 60 minutes Extracted. After the extraction is completed, the extract is separated into solid and liquid, and the separated solution is concentrated under reduced pressure. Then, sucrose fatty acid ester corresponding to 1.0% of the solid content is added and mixed, and 1.25 kg dry extract is obtained by spray drying. Got. After 10 kg of glue (powder cake), lactose 3.7125 kg and magnesium stearate 0.0375 kg are added to 1.25 kg of the dried extract and mixed, and the mixture is made into tablets by a tableting machine by a conventional method. After pulverization, sizing and sieving, a good granule was obtained.

  This granule 15 g contains 1.25 g of herbal extract and 10 g of powdered koji, and is taken in 15 to 3 times a day according to the symptoms.

  The adrenomedullin production enhancer of the present invention can effectively promote the production of adrenomedullin having various physiological activities such as blood flow increasing action and anti-inflammatory action, and is excellent in sustainability and safety of the effect. Therefore, the adrenomedullin production enhancer of the present invention can be used for the treatment of various diseases, such as inflammatory bowel diseases such as Crohn's disease, myocardial disorder, pulmonary hypertension, bone disorder, uterine muscle contraction, dysuria, It is effective for the prevention and treatment of diseases such as bacterial inflammatory diseases and hepatitis.

Claims (10)

Compound represented by the following formula (1)
(Shown wherein ^R 1, ^{R 2} and ^{R 3} are the same or different and each represents a hydrogen atom, a hydroxyl group, -O-Glc, -O-Glc -Glc, the -O-Glc-Ara, or -O-Glc-Rha. (However, Glc represents a glucose residue, Ara represents an arabinose residue, and Rha represents a rhamnose residue.)
And a compound represented by the following formula (2) and / or a compound represented by formula (3)
(In the formula, R ⁴ represents a hydrogen atom or a hydroxyl group, m is 1 or 2, and a wavy line indicates a Z-configuration or an E-configuration)
(Wherein n is 4, 6 or 8)
Prevention or treatment of a disease or disorder selected from the group consisting of hepatitis, dysuria, uterine muscle contraction, bone disorder, pulmonary hypertension, non-bacterial inflammatory disease and myocardial disorder Agent.   The compound represented by the formula (1) is ginsenoside Rb1, ginsenoside Rb2, ginsenoside Rc, ginsenoside Rd, ginsenoside Re, ginsenoside Rg1, ginsenoside Rg2 and ginsenoside Rh1, or one or more ginsenosides The prophylactic / therapeutic agent according to claim 1, which is a kind.   The compound represented by the above formula (2) is composed of α-sanshool, β-sanshool, γ-sanshool, hydroxy-α-sanshool, hydroxy-β-sanshool and hydroxy-γ-sanshool. The prophylactic / therapeutic agent according to claim 1 or 2, which is one or more sanshools selected from the group consisting of:   The compound represented by the formula (3) is one or more compounds selected from the group consisting of 6-shogaol, 8-shogaol and 10-shogaol. The preventive / therapeutic agent according to any of the above sections.   Effective herbal medicine containing the compound represented by the above formula (1) and herbal medicine containing the compound represented by the above formula (2) and / or herbal medicine containing the compound represented by the above formula (3). Prophylactic / therapeutic agent contained as an ingredient.   The preventive / therapeutic agent according to claim 5, wherein the herbal medicine containing the compound represented by the formula (1) is carrot.   The prophylactic / therapeutic agent according to claim 5 or 6, wherein the herbal medicine containing the compound represented by the formula (2) is yam.   The prophylactic / therapeutic agent according to any one of claims 5 to 7, wherein the crude drug containing the compound represented by the formula (3) is psoriasis.   Contains a crude drug containing a compound represented by the above formula (1), a crude drug containing a compound represented by the above formula (2) and / or a crude drug containing a compound represented by the above formula (3). A prophylactic / therapeutic agent for a disease or disorder selected from the group consisting of hepatitis, dysuria, uterine muscle contraction, bone disorder, pulmonary hypertension, non-bacterial inflammatory disease and myocardial disorder, which contains the Kampo formula as an active ingredient.   The prophylactic / therapeutic agent according to claim 9, wherein the Kampo prescription is Daikenchuto.