JP2772060B2 - Gastrointestinal drug - Google Patents

Description

Description: TECHNICAL FIELD The present invention relates to a gastrointestinal drug using a medicinal carrot tissue culture.

(Prior art) Medicinal carrots, especially Panax ginseng
CAMeyer) is widely used in Chinese medicine.
As the medicinal properties, tonic and longevity are known in the old days,
Currently used for gastrointestinal disorders, diabetes, stress, etc. Among various medicinal effects of medicinal carrots, the effects on the gastrointestinal tract include, for example, the Proceedings of International Gi
nseng Symposium (1975) 119-127 describes that it promotes small intestinal motility. However, when the inventor conducted a test on the intestinal motility of a natural medicinal carrot, it was found that the action was extremely weak. In general,
It is known that the intestinal tract relaxes when a medicinal carrot extract is applied to the intestinal tract in vitro. Therefore, the carrot extract contains a substance that relaxes the intestinal tract, which may have a negative effect on the action of promoting intestinal motility when the carrot extract is administered to a living body. Conceivable. This intestinal relaxation action has been studied in detail for ginsenoside, a medicinal carrot extract, and it is known that panaxantriols in medicinal carrots are stronger than panaxanediols ( Arzneim.-Forsch., 25 , 539
~ 547 (1975)). However, the relationship between intestinal relaxation and stimulatory effect on the fraction of the components contained in medicinal carrots and on the resulting single component or when several components are combined has not been investigated. In addition, the detailed effects on the gastrointestinal tract of medicinal carrot components are almost unknown. On the other hand, since medicinal carrots are natural products, their components fluctuate depending on natural conditions such as climate, weather, and soil quality, and the effects on the small intestinal motility also vary greatly. In addition, medicated carrots require a long period of cultivation, so they cannot be produced and used in large quantities as gastrointestinal drugs.

(Problems to be Solved by the Invention) The present invention is intended to solve the above-mentioned conventional drawbacks, and an object of the present invention is to provide a gastrointestinal drug having a high small intestinal motility derived from a medicinal carrot and having a stable quality. It is to provide a gastrointestinal drug which can be produced effectively. It is another object of the present invention to provide a medicinal carrot-derived gastrointestinal drug having a low intestinal relaxing action and a high intestinal motility promoting action.

(Means for Solving the Problems) The inventor has found that a tissue culture of medicinal carrots significantly promotes small intestinal motility as compared with natural medicinal carrots, and has completed the present invention.

The gastrointestinal drug of the present invention contains a medicinal carrot tissue culture or an extract from the tissue culture, thereby achieving the above object.

In a preferred embodiment, the extract is a mixed extract of a medicinal carrot tissue culture and a water-soluble organic solvent, wherein the weight ratio of panaxanetriol to panaxanediol in the extract is 1.4 or less. Yes, the above objective is achieved.

As the medicinal carrot used in the present invention, any medicinal carrot generally used for Chinese herbal medicine or the like can be used. For example, Panax ginseng (Panax
ginseng CAMeyer), Panax Japonic
us), American carrot (Panax Quinquefolium L.),
Ginseng (Panaxnotoginseng (Burk.) FHChen), Himalayan ginseng (Panax pseudo-ginseng Wall.subsp.him
alaicus Hara), ginseng (Panax japonics CAMeyer v)
ar.major (Burk.) CYWu et KMfeng)
anax zingiberenesis CYWu et KM Feng), Sanachi Fangbe (Panax Stipuleanatus Tsai et Feng), and Takeshi Nasaha (Panax Japonics CAMeyer var.angustifolitus)
(Burk.) Cheng et Chu).

The medicinal carrot tissue culture used in the present invention is:
Callus derived from medicinal carrots or differentiated tissues such as root organs obtained by differentiation from the callus. Such a tissue culture is obtained by a usual method. For example, plants such as roots, stems, leaves, and seeds of medicinal carrots are cut out, sterilized with alcohol or the like, and callus is induced by a normal plant tissue culture method. Culture conditions need not be exceptional. As the medium, Murashige-Skoog medium, White medium, Rinsmeyer-Skoog medium, Gausslet medium, Heller's medium, Gamborg's medium, and modified mediums thereof, which are commonly used for plant tissue culture, are used. sell. In particular, Murashige-Skoog medium is preferably used. To this, plant hormones (auxins, cytokinins, etc.) are added as needed to further promote callus induction. Auxins include, for example, 2,4-dichlorophenoxyacetic acid (2,4-D), indoleacetic acid (IAA), indole3-butyric acid (IBA), and naphthaleneacetic acid (NAA). Cytokinins include, for example, kinetin and benzyladenine (BA). Usually, auxins
Cytokinins are added to the medium at a rate of 0.1 to 10 ppm at a rate of 0.01 to 5 ppm. Usually, when cultured at 15 to 35 ° C in the dark, callus is formed on the tissue section after 10 to 30 days. This callus is transferred to a solid medium such as an appropriate agar medium and grown to an appropriate amount. Further, if necessary, the callus is transferred to a solid medium or a liquid medium, and the culture is continued. As this medium, the same medium as described above can be used, and Murashige-Skoog's medium is particularly preferably used. The cultivation is preferably performed in a dark place, and a culturing temperature of 20 to 25 ° C can be used.

It is particularly preferable from the viewpoint of pharmacological action that a root organ (induced root) that can be formed at the time of inducing or culturing the callus is grown by gas phase culture.

The gas phase culture is performed using, for example, a gas phase culture apparatus shown in FIG. This culturing apparatus has a culturing tank 1, which is provided with a heat sterilizing and temperature controlling means 11 outside. The upper part of the tank 1 is provided with an implantation port 100 and a vent 101 for feeding a tissue culture (the above-mentioned root organ), and the lower part is provided with a culture solution container 12. A perforated plate 13 is arranged in the upper gas phase of the storage section 12. The perforated plate 13 holds the culture, and includes a mesh structure such as a stainless steel wire mesh. The size of the hole is set to such a size that the culture does not drop into the culture solution storage unit 12. Perforated plate
A nozzle 15 opens above the culture 14 on 13. The nozzle 15 is provided to supply the culture medium 14 in the storage unit 12 to the culture 14 by a circulation pump 17 via a liquid supply pipe 16 connecting the nozzle 15 and the culture solution storage unit 12. A plurality of perforated plates 13 may be provided in parallel in the culture tank 1 at intervals. When a plurality of perforated plates 13 are installed, nozzles 15 are respectively opened above the cultures 14 on each perforated plate 13,
It is preferable that the required minimum amount of the culture solution is evenly supplied to the culture 14.

In order to carry out the vapor phase culture, first, the culture is put into the culture tank 1 from the implantation port 100 of the apparatus, and the culture is placed on the porous plate 13 almost uniformly. The culture medium is sprayed from the nozzle 15 onto the culture 14 through the liquid feed pipe 16 by the pump 17. Air or oxygen is supplied from one of the vents 101, and is discharged from the local vent 101.

By conducting such a gas phase culture for about 21 to 35 days, the culture grows.

The callus and / or root organ obtained by the above culture are collected by appropriate means such as filtration. It is dried slowly on the sun or under warm air at 30-70 ° C. This dried culture can be directly used as a gastrointestinal drug by pulverizing it into granules or powder having a desired particle size using an ordinary pulverizer. The particle size of this powder is usually 5 to 100
μm. The dosage of the above granules and powder is 1 to 1 per day.
10g is appropriate. Alternatively, an extract is obtained from the culture or the dried culture using water or a water-water-soluble organic solvent. As the water-soluble organic solvent, ethanol, methanol, acetone and the like can be used.
The concentration of the water-soluble organic solvent may be in the range of 0 to 100%, and the amount of the extract to be used is three times that of the culture (as dry matter).
A volume of up to 20 times is appropriate.

The weight ratio between panaxanetriol and panaxanediol in the extract thus obtained is 1.4 or less, preferably 0.8 to 1.0. When this value exceeds 1.4, the intestinal relaxation action increases when the extract is administered.
A desired intestinal motility promoting effect may not be obtained. In particular, when an extract in which the weight ratio of panaxanetriol to panaxanediol is in the range of 0.8 to 1.0 is used, the intestinal relaxation action is low and the exercise promoting action is high.

The extract in which the weight ratio of panaxanetriol to panaxanediol is 1.4 or less can be obtained, for example, by selecting a strain of an appropriate culture. The strains of the culture are selected by measuring the content ratio of panaxantriol and panaxanediol in the culture during tissue culture. The contents of panaxantriol and panaxanediol were determined by liquid chromatography,
It can be measured by thin layer chromatography, chromatography using other appropriate columns, or the like. Even when the weight ratio of the extract from the medicinal carrot culture exceeds 1.4, the weight ratio can be adjusted to 1.4 or less by adding panaxanediol to the extract.

The obtained extract can be taken as it is or after removing the solvent. Considering ease of taking and portability, it is preferable that the extract is spray-dried or vacuum freeze-dried to obtain a dry extract powder.
Further, this dry extract powder can be formulated and used as a gastrointestinal drug. For example, powders, granules, tablets, capsules, and the like can be prepared by adding a suitable excipient, auxiliary, and the like used in ordinary preparations to a dry extract powder according to a conventional method. When the above extract or a preparation obtained from the extract is used, the dose is 1 to 20 g per time in terms of dry culture. The gastrointestinal drug of the present invention
It is usually appropriate to take the drug three times a day according to the symptoms. The gastrointestinal drug of the present invention does not show toxic effects on living organisms and is extremely safe, as can be seen from the examples described below.

(Examples) The present invention will be described with reference to the following examples.

Example 1 Callus was induced from roots of Panax ginseng (commonly known as Korean ginseng) by an ordinary method. Callus was induced by culturing under a normal culture condition in a solid MS medium containing agar (Murashige-Skoog medium). 100g of above callus
(Wet weight) was used as a seed, aseptically inoculated in a gas phase culture apparatus shown in FIG. 1, and the culture solution was continuously supplied to the seed tissue at 25 ° C. at a rate of 100 ml / min. As a culture solution, an MS medium containing 0.1 ppm of kinetin and 2 ppm of indolebutyric acid was used.
After culturing for 4 weeks, when the culture was harvested, the wet weight was 583 g.
Met. The culture was dried with warm air at 40 ° C for 72 hours, and 36.
7 g of dried product was obtained. 20 g of the dried product was extracted with 400 ml of 50% ethanol and freeze-dried to obtain 8 g of an extract powder.

Experimental Example 1 [Single Administration Test] Four-week-old ddY male mice (body weight 25 to 30 g) were grouped into groups 5 to 5
Ten animals were fasted for 24 hours before administration of the test substance. As test substances, the extract powder obtained in Example 1, the extract powder obtained from commercially available natural panax ginseng 6-year-old roots (Korean, Japanese or Chinese) and freeze-dried as in Example 1 or carbachol ( Control) was used. These test substances were orally administered at a dose of 400,800,2000 mg / kg (subcutaneous administration for control carbachol), and then 0.3 ml of a 5% aqueous carbon powder suspension was orally administered. Twenty minutes later, they were sacrificed and their small intestine was removed. After fixing the isolated small intestine with formalin, the distance from the pyloric part of the stomach to the moving tip of the carbon powder was measured, and the small intestine transport ability was calculated by the following equation.

Small intestinal transport capacity = (distance from gastric pylorus to tip of carbon powder suspension transfer / length from gastric pylorus to cecum) × 100 Table 1 shows small intestinal transport capacity when each test substance was administered.

From the results in Table 1, the extract powder from the cultured carrot was
It can be seen that small intestinal motility is significantly promoted. On the other hand, no small intestinal motility is observed even when an extract powder from a natural panax ginseng root is used.

Experimental Example 2 [Continuous administration test] Four-week-old ddY male mice (body weight 25 to 30 g) were grouped in groups 5 to 5
The test substances used in Experimental Example 1 were 400 and 80, respectively.
Oral administration was performed daily at a dose of 0.2000 mg / kg for two consecutive weeks (subcutaneous administration for control carbachol). After the final administration, these mice were fasted for 24 hours, and the intestinal transport ability was evaluated using the aqueous carbon powder suspension according to the method described in Example 1. Table 2 shows the results.

Experimental Example 3 The extract powder obtained in Example 1 was applied to ddY at a dose of 5 g / kg.
There were no deaths in an acute toxicity test after oral administration to 10 male male mice. Thus, the gastrointestinal drug of the present invention has extremely low toxicity and high safety.

Example 2 200 g of dry extract powder produced according to the method of Example 1 was mixed with 20 g of microcrystalline cellulose and magnesium stearate 5
g), and press the mixture with a single-shot tableting machine.
Tablets with a diameter of 7 mm and a weight of 225 mg were produced. One tablet contains 200 mg of the dried extract of Panax ginseng. As for this tablet, 5 to 50 tablets per day should be used according to the symptoms.
Take in divided doses.

Example 3 500 mg of dry extract powder produced by the method of Example 1
Were filled into hard capsules. This capsule is taken 2 to 40 capsules a day in 3 divided doses according to the symptoms.

Example 4 A culture was obtained from Panax ginseng in the same manner as in Example 1. A 50% ethanol extract was obtained from the obtained culture, and the extract was subjected to high performance liquid chromatography to measure the content of panaxanetriol and panaxanediol. A culture strain corresponding to the extract having a weight ratio of panaxantriol to panaxanediol of 1.4 or less was selected and further grown. To 50 g of this culture (dry matter) was added 50% ethanol of 9 and the mixture was heated at 70 ° C. for 1 hour, and the extract was collected by filtration. Next, 9% 50% ethanol was added to the residue, and the mixture was heated at 70 ° C. for 1 hour, and the extract was collected by filtration. The obtained extracts were combined and the solvent was distilled off under reduced pressure. 206.8 g of an extract (extract) having a weight ratio of panaxantriol / panaxanediol of 0.92 was obtained.

Experimental Example 4 [Assay of Intestinal Relaxing Action] The small intestinal tract of a 6-week-old male ddY mouse (body weight 25 to 30 g) was excised, and the intestinal relaxing action was assayed by the following method. As the test substance, the extract obtained in Example 4 or commercially available natural ginseng A to E (A: Korean carrot root; B: Japanese carrot root; C: Chinese carrot root; D: Chinese carrot root (E: Chinese carrot root) and an extract obtained in the same manner as in Example 4. Assay method: One end of the isolated intestinal tract is fixed in a Tyrode solution (27 ° C.) aerated with 95% O ₂ at a rate of 20 to 30 bubbles / min. After connecting the other end of the intestinal tract to the displacement transducer, acetylcholine is added to contract the small intestine, which is washed and relaxed. By performing this operation several times, the degree of contraction of the intestinal tract is adjusted to be constant. Then add 10 ^-4 g / ml of the test substance and measure the relaxed length of the small intestine.

As described above, in the assay method in which contraction / relaxation was measured while keeping the tension constant, the relaxation effect was greatest when natural ginseng A was added. The relaxed length (length after relaxation-original length) is defined as the relaxation rate of 100% (the relaxation rate of the original length is 0%), and the intestinal relaxation rate (%) of each test substance. Was calculated. Table 3 shows the results.

Experimental Example 5 [Single Administration Test] Four-week-old male ddY mice (body weight 25 to 30 g) were grouped in groups 5 to 5
10 animals were fasted. As a test substance, commercially available extracts of natural carrots F to H (F: Korean carrot root; G: Japanese carrot root; H: Chinese carrot root) obtained by the same method as in Example 4 or carbachol (Control) was used. These test substances were orally administered at the doses shown in Table 4 (subcutaneous administration for control carbachol), and 30 minutes later, 0.3 ml of a 5% carbon powder aqueous suspension was orally administered. Twenty minutes after administration, the mice were sacrificed and the small intestine was removed. After fixing the isolated small intestine with formalin, the distance from the pyloric part of the stomach to the moving tip of the carbon powder was measured.
The small intestine transport ability was calculated in the same manner as in Experimental Example 1.

Table 4 shows the small intestinal transit ability when each test substance was administered. The dose of the extract in the table indicates the total amount of saponin in the extract.

Example 5 A culture was obtained from Panax ginseng in the same manner as in Example 1. From this culture, using the same solvent as in Example 4,
218.0 g of the extract was obtained. The weight ratio of panaxanetriol to panaxanediol in the obtained extract was 1.74. Ginsenoside b1 (a type of panaxanediol) was added to this extract at a rate of 9.55 mg / g, and the weight ratio of panaxanetriol to panaxanediol was determined.
Adjusted to 0.9.

Example 6 The extract obtained in Example 4 in which the weight ratio of panaxanetriol and panaxanediol was adjusted was freeze-dried to obtain 250 g of an extract powder. Dry extract powder 2 obtained
00 g was mixed with 20 g of microcrystalline cellulose and 5 g of magnesium stearate, and the mixture was tableted with a single-shot tableting machine to produce tablets having a diameter of 7 mm and a weight of 225 mg. This tablet 1
Tablets contain 200 mg of dried panax ginseng powder. These tablets are to be taken in 5 to 50 tablets a day in 3 divided doses according to the symptoms.

Example 7 Hard capsules were filled with 500 mg each of the dry extract powder obtained in Example 6. This capsule is taken 2 to 40 capsules a day in 3 divided doses according to the symptoms.

(Effect of the Invention) According to the present invention, a gastrointestinal drug derived from a culture of medicinal carrots, which significantly promotes small intestinal motility, is thus obtained. By obtaining an extract in which the weight ratio of panaxanetriol to panaxanediol in the extract from the culture is within a predetermined range, an effective gastrointestinal drug having a further lower intestinal relaxation action is provided. Such small bowel motility is rarely found in natural medicinal carrots. Since the gastrointestinal drug of the present invention utilizes a culture of medicinal carrots, it can be produced in large quantities without being affected by natural conditions such as season, weather, and soil quality.
Has a stable pharmacological effect.

[Brief description of the drawings]

FIG. 1 is a front sectional view showing an example of an apparatus for vapor-phase cultivating a Panax ginseng culture used for a gastrointestinal drug of the present invention. 1 ... culture tank, 11 ... heat sterilization and temperature control means, 12 ...
... Culture buffer, 13 ... Perforated plate, 14 ... Culture, 15 ... Nozzle, 16 ... Send tube, 17 ... Circulation pump, 100 ... Injection port, 1
01 …… Vents.

──────────────────────────────────────────────────続 き Continuation of the front page (56) References JP-A-54-135210 (JP, A) JP-A-61-18722 (JP, A) JP-A-61-209599 (JP, A) JP-A-60-1985 89496 (JP, A) JP-A-62-39529 (JP, A) JP-A-62-56437 (JP, A) JP-A-62-71501 (JP, A) JP-A-62-71502 (JP, A) JP-A-2-234696 (JP, A) Akamatsu Kaneyoshi, "New Revised Japanese Medicine," Medical and Dental Medicine Publishing Co., Ltd., October 15, 1980, 1st edition, 5th printing, pp. 203-6 (58) Field surveyed (Int.Cl. ⁶ , DB name) A61K 35/78 M

Claims (3)

(57) [Claims] 1. A gastrointestinal drug for promoting small intestinal motility, comprising a tissue culture of a medicinal carrot or an extract from the tissue culture. 2. The extract is a mixed extract of a medicinal carrot tissue culture and a water-soluble organic solvent, wherein the weight ratio of panaxanetriol to panaxanediol in the extract is 1.4 or less. A gastrointestinal drug according to claim 1. 3. The method according to claim 2, wherein the tissue culture in which the weight ratio of panaxantriol to panaxanediol in the extract is 1.4 or less is obtained by selecting a culture strain of medicinal carrot. Gastrointestinal drug.