JP2000034233A - Inhibitor of production of nitric oxide - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain an inhibitor having inhibiting actions on the production of nitric oxide by making the inhibitor include a specific folkloric medicine in Indonesia or its extract as an active ingredient. SOLUTION: This inhibitor is obtained by including the following at least one or more kinds of plant bodies themselves used as folkloric medicines in Indonesia (botanical name, Indonesian name) or an extract of the plants as an active ingredient: Andrographis paniculata, Sambi, boto; Caesalpinia sappan, Kaya secang; Schima wallichii, Buah cangkok; Alstonia scholaris, Babakan pule; Graptophyllum pictum, Daum wungu; Usnea spp., Akar angin; Rheum officinale, Klmebak; Sindora javanica, Saparantu; Vitex trifolia, Legundi; Anacardium occidentale, Daun jambu mete; Gumnopetalum leucosticum, Duri kemarung; Equisetum debile, Greges otot; Kyllinga monocephala, Akar teki; Kyllinga vrevifolia, Akar teki; Ardisia fuliginosa, Ajag; Entada phaseoloides, Tariyu and Alyxia reinwardti, Kayu polo sari.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to a nitric oxide production inhibitor comprising a specific plant or an extract thereof as an active ingredient.

[0002]

BACKGROUND OF THE INVENTION In the early 1980's, during the study of nitrogen oxides in vivo, it was first discovered that nitric oxide (hereinafter sometimes abbreviated as NO) was produced in vivo. Following that discovery, NO attracted the attention of many researchers, and in 1987, it was reported that NO was the main body of vascular endothelial cell-derived relaxing factor. At present, the relationship between NO physiological functions and disease states has been elucidated in a wide range of fields such as circulation, immunity, and nervous system. Among them, for example, it has been elucidated that NO constantly produced in the body plays an important role in maintaining hemodynamic homeostasis. On the other hand, in sepsis, it is said that a large amount of NO is produced by the action of cytokines activated by endotoxin, and this causes an endotoxin shock state such as endothelial cell damage and reduced myocardial contractility.

Conventionally, prostacyclin has been known as an endogenous physiologically active substance having an inhibitory effect on vasorelaxation and platelet aggregation, but another new vasorelaxant was discovered in 1980 by Furchgott et al. Was discovered as a factor (Endothelium Derived Relaxing Factor, EDRF), and in 1987 Monc was found to be identical to NO.
ada et al. and Ignarro et al.
gnarro, LJ: Endothelium-derived nitric oxide: a
ctions and properties.FASEB J. 3, 31-36,1989., Kn
owles, RGand Moncada, S.: Nitric oxide asa signal i
n blood vessels. Trends Biochem. Sci. 17, 399-402,199
2., Moncada, S., Palmer, RMJ and Higgs, EA: Nit
ric oxide: physiology, pathophysiology and pharmac
ology.Pharmacolog. Rev. 43, 109-142, 1991.).

[0004] Since then, the role of NO as a signal transmitting substance in many physiological functions in the living body, such as the nervous system and the immune system, other than the circulatory system, has been revealed one after another. That is, in the nervous system, it acts as a neurotransmitter,
It is involved in the establishment of synaptic plasticity (long-term cerebellar depression and long-term hippocampal phenomena) (Katsuei Shibuki: NO and plasticity of the nervous system-regulation of long-term depression of the cerebellum. Experimental Medicine 11,2451-2456,1
993.) Furthermore, in the immune system, the importance of NO produced from macrophages acting as effector cells has been pointed out in the host defense mechanism against tumor cells and pathogens (Takio Taki, Masayasu Nakano: Arginine and macrophage in macrophages) Antibacterial and antitumor effects of its metabolites. History of medicine 156,194-197,1991.). Even if it is limited to vasodilator action, it has been proved that it acts not only from the intima side as EDRF but also acts from the outer membrane side as a non-adrenergic / non-cholinergic vasodilator neurotransmitter It has been pointed out that NO regulates various physiological functions by a complicated mechanism.

[0005] NO, which has been found to be produced in vivo as described above, is produced by nitric oxide synthase (NOS) using L-arginine as a substrate. NOS has at least non-inducible (vascular endothelial and neural) and inducible isozymes. Endothelial NOS
Is mainly present in vascular endothelial cells, and its activity is controlled by the intracellular calcium concentration. Neuronal NOS is present in central nerve cells, peripheral nerve cells, or pancreatic islet β-cells, gastrointestinal nerves, adrenal medulla, renal compact plaques, etc.
Similar to S, the activity is controlled by the intracellular calcium concentration.

[0006] Endothelial NOS and neural NOS (co
nstitutive NOS, abbreviated as c-NOS) is constantly present in cells, and there is almost no change in the amount of enzymes due to physiological changes. Inducible NOS (abbreviated as inducible NOS, i-NOS) includes hepatic parenchymal cells, neutrophils, macrophages, smooth muscle, fibroblasts, renal mesangial cells, gastrointestinal epithelium, pancreatic islet β cells, vascular smooth muscle cells Or it exists in glial cells and the like. This is not usually observed in cells, but is induced by stimulation with endotoxin, various cytokines, and the like.

[0007] The action of NO produced by NOS is diverse and includes, for example, vasorelaxant action, platelet aggregation inhibitory action,
Adhesion suppression, leukocyte adhesion / migration suppression, sympathetic nerve activity suppression,
Endotoxin shock, damage due to hypotension due to endotoxin / cytokine, action as a signal transmitter between nerve cells, ischemic brain cell damage, antitumor, bactericidal action,
Autoimmune diseases, insulin-dependent diabetes mellitus, arthritis, tissue damage after transplantation, rejection and the like.

[0008] In analyzing the physiological activity of NO in vivo, NO synthase inhibitors are useful and may be used as therapeutic agents for shock and ischemic diseases. Development of NOS inhibitors is currently underway. For example, there is an arginine analog as a substrate competitor, and Nω-monomethyl-L-arginine (L-NMM
A), Nω-nitro-L-arginine (L-NNA),
Nω-amino-L-arginine (L-NAA), Nω-
Iminoethyl-ornithine (L-NIO) and the like correspond thereto.

[0009] However, most of the known NOS inhibitors including the above representative examples inhibit not only iNOS but also cNOS. Even the regulation of circulatory dynamics is suppressed, and it is not possible to avoid side effects such as an increase in blood pressure and a decrease in organ blood flow. In addition, there is a concern that the use of these methods may affect the central nervous system and impotence.
As described above, conventionally known NOS inhibitors cannot be evaluated as pharmaceuticals.
There has been a demand for the development of a drug having an NO production inhibitory action capable of selectively inhibiting iNOS.

[0010]

Means for Solving the Problems The present inventors have developed iNOS.
As a result of diligent studies to find a drug having an NO production inhibitory action capable of selectively inhibiting NR, it was found that a specific Indonesian folklore drug and its extract have an NO production inhibitory action, completed.

That is, the present invention provides a scientific name: Andrographis paniculata Indonesian name (Sambi boto) Scientific name: Caesalpinia sappan Indonesian name (Kaya secang) Scientific name: Schima wallichii Indonesian name (Buah cangkok) Scientific name: Alstonia scholaris Indonesian name (Babakan pule) Graptophyllum pictum Indonesian name (Daum wungu) Scientific name: Usnea spp. Indonesian name (Akar angin) Scientific name: Rheum officinale Indonesian name (Klembak) Scientific name: Sindora javanica Indonesian name (Saparantu) Scientific name: Vitex trifolia Indonesian name (Legundi) occidental identium Anacard Indonesian name (Daun jambu mete) Scientific name: Gymnopetalum leucosticum Indonesian name (Duri kemarung) Scientific name: Equisetum debile Indonesian name (Greges otot) Scientific name: Kyllinga monocephala Indonesian name (Akar teki) Scientific name: Kyllinga brevifolia Indonesian name (Akar t) fuliginosa Indonesian name (Ajag) Scientific name: Entada phaseoloides Indonesian name (Tariyu) and scientific name: Alyxia reinwardti At least one plant selected from the group consisting of Indonesian name (Kayu polo sari), or by a supercritical extraction method using a plant solvent extract or carbon dioxide gas An NO production inhibitor characterized by containing an extracted extract as an active ingredient.
The present invention also relates to the whole name of Andrographis paniculata Indonesian name (Sambi boto) Scientific name: Caesalpinia sappan Indonesian name (Kaya secang) Wood part Scientific name: Schima wallichii Indonesian name (Buah cangkok) Real scientific name: Alstonia scholaris Indonesian name (Babakan pule) bark Scientific name: Graptophyllum pictum Indonesian name (Daum wungu) whole plant Scientific name: Usnea spp. Indonesian name (Akar angin) whole plant Scientific name: Rheum officinale Indonesian name (Klembak) Root Scientific name: Sindora javanica Indonesian name (Saparantu) Scientific name: Vitex trifolia Indonesian name (Legundi) Leaf Scientific name: Anacardium occidentale Indonesian name (Daun jambu mete) Leaf Scientific name: Gymnopetalum leucosticum Indonesian name (Duri kemarung) Thorn Scientific name: Equisetum debile Indonesian name (Greges otot) Stalk Scientific name: Kyllinga monocephala Whole plant of Indonesian name (Akar teki) Scientific name: Kyllinga brevifolia Indonesian name (A kar teki) whole plant Scientific name: Ardisia fuliginosa Indonesian name (Ajag) whole plant Scientific name: Entada phaseoloides Indonesian name (Tariyu) stem / leaf and scientific name: Alyxia reinwardti Indonesian name (Kayu polo sari) stem A nitric oxide production inhibitor characterized by containing at least one or more plants themselves, or an extract extracted by a supercritical extraction method using a solvent extract of plants or carbon dioxide as an active ingredient. . The invention further provides that the solvent is chloroform, methanol, ethanol, propylene glycol and 1,3
A NO production inhibitor, which is at least one member selected from the group consisting of butylene glycol.

The above-mentioned plant derived from Indonesia, which has been shown to be effective in suppressing NO production according to the present invention, has been reported as an ancient folklore, and is derived from Jamu (like Chinese herbal medicine in China). Used as a constituent. However, it has never been known that these plants, their extracts, and jams containing them as components have NO production inhibitory action.

[0013]

DESCRIPTION OF THE PREFERRED EMBODIMENTS The present invention will be described below in detail. Ingredients preferably used in the present invention Scientific name: Andrographis paniculata Indonesian name (Sambi boto) Whole plant Scientific name: Caesalpinia sappan Indonesian name (Kaya secang) Kibe Scientific name: Schima wallichii Indonesian name (Buah cangkok) Real scientific name: Alstonia scholaris Indonesian name Bark of (Babakan pule) Scientific name: Graptophyllum pictum Whole plant of Indonesian name (Daum wungu) Scientific name: Usnea spp. Whole plant of Indonesian name (Akar angin) Scientific name: Rheum officinale Root of Indonesian name (Klembak) Scientific name: Sindora javanica Indonesian name (Saparantu) Scientific name: Vitex trifolia Indonesian name (Legundi) Leaf Scientific name: Anacardium occidentale Indonesian name (Daun jambu mete) Leaf Scientific name: Gymnopetalum leucosticum Indonesian name (Duri kemarung) Thorn Scientific name: Equisetum debile Indonesian name (Greges) ) Stem Scientific name: Kyllinga monocephala Indonesian name (Akar teki) Scientific name: Kyllinga brevifo lia Whole plant of Indonesian name (Akar teki) Scientific name: Ardisia fuliginosa Whole plant of Indonesian name (Ajag) Scientific name: Entada phaseoloides Stem and leaf of Indonesian name (Tariyu) Scientific name: Alyxia reinwardti Dry name of Indonesian name (Kayu polo sari) Is also available in other markets, including the Indonesian market.

Although the extracted plant of the active ingredient may be used without being cut as it is, it is more preferable to use the cut plant before the extraction operation in that the extraction time can be shortened. As a method for extracting the active ingredient from the raw materials, a known method can be employed, and examples thereof include a supercritical extraction method using carbon dioxide gas and an extraction method using a solvent. Examples of the extraction method using a solvent include a method in which a solvent is added to a raw material and heat treatment is performed at a reflux temperature of the solvent. In general, this heat treatment is preferably performed at a temperature of 80 ° C. or lower, and is performed under reflux using a known extraction device.
An extract can be obtained by heating for a time. Alternatively, an extract can be obtained by digesting the dry powder of the raw material in a solvent. Supercritical fluid is
Since its dissolving power can be easily and continuously controlled by pressure and temperature over a wide range, the supercritical extraction method utilizing this property is expected as a new separation method. In particular, supercritical carbon dioxide (SC-CO ₂ ) has a relatively low critical point (critical temperature 304.2 K, critical pressure 72.8 atm), and thus can be applied to thermally unstable substances. The supercritical extraction method and the extraction method using a solvent may be used in combination. These extraction operations can be repeatedly performed on the raw material residue after the first extraction operation. The amount of solvent used for extraction is 100 per 100 parts by weight of raw material.
The suitable amount is from 10 to 10,000 parts by weight, more preferably from 300 to 5,000 parts by weight.

Examples of the extraction solvent usable in the present invention include water, lower alcohols such as methanol, ethanol, propyl alcohol, isopropyl alcohol, butanol and isobutanol, and polyhydric alcohols such as propylene glycol and 1,3-butylene glycol. Acetone, dioxane, methyl ethyl ketone, acetonitrile, ethyl acetate, butyl methyl ketone, diethyl ether, dichloromethane, xylene, trichloroethylene, carbon tetrachloride, benzene, chloroform, toluene and the like can be mentioned. Particularly, chloroform, methanol, ethanol, propylene glycol, and 1,3-butylene glycol are preferred. The above solvents may be used alone, or two or more kinds may be used as a mixture, or water and an organic solvent may be used in combination. When the lower alcohol and the polyhydric alcohol are used as hydroalcohols, the water content is preferably 50% or less. The extract thus obtained is concentrated under reduced pressure, and then dried to obtain the extract as a powder. Also, depending on the solvent used, it is possible to use the extract as it is as an active ingredient of the NO production inhibitor, for example, ethanol,
An extract with propylene glycol, 1,3-butylene glycol or the like may be used without removing the solvent.

The supercritical extraction method is an extraction technique that has attracted attention in a wide range of fields such as the food, chemical, pharmaceutical, and cosmetic industries. In particular, when carbon dioxide is used as an extractant, the critical point (75 kg / cm ² , 31 ° C.) is relatively low and the safety is high, so that components unstable to heat and highly volatile components are used. Can be efficiently extracted and separated. Extraction of a target component from a raw material can be achieved by setting the pressure and temperature of carbon dioxide at or above a critical point. Generally, at the same temperature, the higher the pressure, the higher the dissolving power of supercritical carbon dioxide. The supercritical carbon dioxide extraction in the present invention can be performed by a method known per se. For example,
Using a supercritical extraction device, the temperature of the high pressure cell section is 33-40.
C., preferably 35.degree. C., a pressure of 75 to 300 atm, preferably 150 atm, and a flow rate of carbon dioxide of 0.5 to 5.0.
dm ³ / min, preferably at a condition of 4. 0dm ³ / min. Thereby, the extract containing the NO production inhibitor can be efficiently collected.

In addition to the extract of the above-mentioned raw materials, which is an active ingredient, the NO production inhibitor of the present invention may be appropriately added to the extract unless it is not harmful to the extract and is not suitable for a product utilizing the NO production inhibitor. It is possible to mix the agent according to a conventional method, and glycyrrhizic acid, dipotassium glycyrrhizinate, lysozyme chloride, lytic enzyme, mutanase, chlorhexidine, sorbic acid, alexidine, hinokitiol, cetylpyridinium chloride, alkyl glycine, alkyl diaminoethyl glycine Salt, allantoin, ε-aminocaproic acid, azulene, vitamin E and derivatives thereof, sodium monofluorophosphate, sodium fluoride, stannous fluoride, water-soluble primary or secondary phosphate, quaternary ammonium compound, Incorporation of active ingredients such as sodium chloride It can be.

Application to Products The NO production inhibitor of the present invention is preferably a scientific name: whole plant of Andrographis paniculata Indonesian name (Sambi boto) Scientific name: Caesalpinia sappan Indonesian name (Kaya secang) Kobe Scientific name: Schima wallichii Indonesia Name (Buah cangkok) Practical name: Alstonia scholaris Bark of Indonesian name (Babakan pule) Scientific name: Graptophyllum pictum Whole plant of Indonesian name (Daum wungu) Scientific name: Usnea spp. Whole plant of Indonesian name (Akar angin) Scientific name: Rheum officinale Indonesia Scientific name of Sindora javanica Indonesian name (Saparantu) Scientific name of Vitex trifolia Indonesian name (Legundi) Leaf Scientific name: Anacardium occidentale Indonesian name (Daun jambu mete) Leaf Scientific name: Gymnopetalum leucosticum Indonesian name (Duri kemarung) Scientific name: Equisetum debile Indonesian name (Greges otot) Stem Scientific name: Kyllinga monocephala Indonesian name (Ak Scientific name: Kyllinga brevifolia Scientific name: Indonesian name (Akar teki) Scientific name: Ardisia fuliginosa Scientific name: Indonesian name (Ajag) Scientific name: Entada phaseoloides Indonesian name (Tariyu) stem / leaf Scientific name: Alyxia reinwardti Indonesian name (Kayu polo sari) stalk selected from one or more Indonesian folklore as it is, or its extract as an active ingredient,
It may be formulated by combining this with a known pharmaceutical carrier.

The NO production inhibitor of the present invention can be administered as an oral agent or a parenteral agent such as an injection or an infusion.

Pharmaceutical carriers can be selected according to the above-mentioned administration forms and dosage forms. In the case of oral preparations, for example, starch, lactose, sucrose, mannitol, carboxymethylcellulose, corn starch, inorganic salts and the like are used. You. In addition, in preparing an oral preparation, a binder, a disintegrant,
Surfactants, lubricants, fluidity promoters, flavoring agents, coloring agents,
Flavors and the like can be blended. Specific examples thereof include the following.

Examples of the binder include starch, dextrin, gum arabic, gelatin, hydroxypropyl starch, methylcellulose, sodium carboxymethylcellulose, hydroxypropylcellulose, crystalline cellulose, ethylcellulose, polyvinylpyrrolidone, macrogol and the like.

Examples of the disintegrant include starch, hydroxypropyl starch, sodium carboxymethylcellulose, calcium carboxymethylcellulose, carboxymethylcellulose, low-substituted hydroxypropylcellulose and the like.

Examples of the surfactant include sodium lauryl sulfate, soybean lecithin, sucrose fatty acid ester, polysorbate 80 and the like.

As the lubricant, for example, talc, waxes,
Examples include hydrogenated vegetable oils, sucrose fatty acid esters, magnesium stearate, calcium stearate, aluminum stearate, polyethylene glycol and the like.

Examples of the fluidity promoter include light anhydrous silicic acid, dried aluminum hydroxide gel, synthetic aluminum silicate, magnesium silicate and the like.

The NO production inhibitor of the present invention can also be administered as oral liquids such as suspensions, emulsions, syrups, elixirs and the like. Agents and coloring agents can be added.

On the other hand, parenteral preparations are prepared according to a conventional method, and are generally used as diluents: distilled water for injection, physiological saline, aqueous glucose solution, vegetable oil for injection, sesame oil, peanut oil, soybean oil, corn oil, propylene oil. Glycol, polyethylene glycol and the like can be used.
Further, if necessary, a bactericide, a preservative, and a stabilizer may be added. Further, from the viewpoint of stability, this parenteral preparation can be frozen after filling into a vial or the like, water can be removed by a usual freeze-drying technique, and a liquid preparation can be prepared from the freeze-dried product immediately before use. Further, if necessary, an isotonic agent, a stabilizer, a preservative, a soothing agent and the like can be appropriately added.

The dose of the NO production inhibitor of the present invention varies depending on the route of administration, the degree of the disease, the age of the recipient, and the like. In general, in the case of oral administration, one species selected from the above plants is used per adult per day. Alternatively, the amount of the dry extract of two or more crude drugs may be about 1 to 10 g in 1 to 3 divided doses.

The various crude drugs used in the present invention have a long history as a component plant of jamu and have been confirmed to be safe, so that they can be used with confidence. For example, oral administration of 15 g / kg, which is the administration limit, to mice and rats is extremely safe as apparent from the fact that no deaths or abnormal findings were observed.

The NO production inhibitor of the present invention can be added to foods, feeds and the like according to a conventional method, and the amount of addition can be appropriately selected according to the type of the product. Is 0.001 to 5% by weight, and more preferably 0.005 to 2% by weight. As a product to which the NO production inhibitor of the present invention is applied, specifically, pasta, chewing gum, chewing jelly,
Foods such as coffee drinks and feeds such as dog foods and cat foods are included.

[0031]

EXAMPLES Next, the present invention will be described in more detail with reference to Production Examples and Examples, but the present invention is not limited to these Examples. Production Example 1 Scientific name: Andrographis paniculata Indonesian name (Sambi
boto) 50 g for 100 g of crushed dried whole plant
/ v% ethanol (1,000 ml)
Stirring extraction was performed for a day. This is centrifuged and filtered under pressure,
An extract was obtained. This extract was freeze-dried to obtain an extract. Production Example 2 Scientific name: Caesalpinia sappan Indonesian name (Kaya secan
g) 50 v / v per 100 g of crushed dried xylem
1,000 ml of ethanol was added thereto, and the mixture was extracted with stirring at room temperature in a dark place for 7 days. This was centrifuged and filtered under pressure to obtain an extract. This extract was freeze-dried to obtain an extract. Production Example 3 Scientific name: Schima wallichii Indonesian name (Buah cangko
k) 1,000 g of 50 v / v% ethanol was added to 100 g of the actually dried crushed product, and the mixture was extracted with stirring at room temperature in a dark place for 7 days. This was centrifuged and filtered under pressure to obtain an extract. This extract was freeze-dried to obtain an extract. Production Example 4 Scientific name: Alstonia scholaris Indonesian name (Babakan p
ule), 50 v / v for 100 g of the crushed dry bark
1,000 ml of ethanol was added thereto, and the mixture was extracted with stirring at room temperature in a dark place for 7 days. This was centrifuged and filtered under pressure to obtain an extract. This extract was freeze-dried to obtain an extract. Production Example 5 Scientific name: Graphtophyllum pictum Indonesian name (Daum wu
ngu), 50 v / v for 100 g of crushed dried whole plant
1,000 ml of ethanol was added thereto, and the mixture was extracted with stirring at room temperature in a dark place for 7 days. This was centrifuged and filtered under pressure to obtain an extract. This extract was freeze-dried to obtain an extract. Production Example 6 Scientific name: Usnea spp. To 100 g of crushed dried whole plant of Indonesian name (Akar angin), 1,000 ml of 50 v / v% ethanol was added, and the mixture was stirred and extracted in a dark place at room temperature for 7 days. Was. This was centrifuged and filtered under pressure to obtain an extract. This extract was freeze-dried to obtain an extract. Production Example 7 Scientific name: Rheum officinale To 100 g of crushed dried roots of Indonesian name (Klembak), 1,000 ml of 50 v / v% ethanol was added, and the mixture was stirred and extracted in a dark place at room temperature for 7 days. This was centrifuged and filtered under pressure to obtain an extract. This extract was freeze-dried to obtain an extract. Production Example 8 Scientific name: Sindora javanica Indonesian name (Saparantu)
1,000 g of 50 v / v% ethanol was added to 100 g of the crushed product of the actual dried product, and the mixture was extracted with stirring at room temperature in a dark place for 7 days. This was centrifuged and filtered under pressure to obtain an extract. This extract was freeze-dried to obtain an extract. Production Example 9 Scientific name: Vitex trifolia To 100 g of crushed dried leaf of Indonesian name (Legundi), 1,000 ml of 50 v / v% ethanol was added, and the mixture was stirred and extracted at room temperature in a dark place for 7 days. This was centrifuged and filtered under pressure to obtain an extract. This extract was freeze-dried to obtain an extract. Production example 10 Scientific name: Anacardium occidentale Indonesian name (Daun
jambu mete)
1,000 ml of 0 v / v% ethanol was added, and the mixture was stirred and extracted in a dark place at room temperature for 7 days. This was centrifuged and filtered under pressure to obtain an extract. This extract was freeze-dried to obtain an extract. Production example 11 Scientific name: Gymnopetalum leucosticum Indonesian name (Dur
i kemarung) 5 g
1,000 ml of 0 v / v% ethanol was added, and the mixture was stirred and extracted in a dark place at room temperature for 7 days. This was centrifuged and filtered under pressure to obtain an extract. This extract was freeze-dried to obtain an extract. Production example 12 Scientific name: Equisetum debile Indonesian name (Greges oto
t) 50 v / v% to 100 g of crushed dried stem
1,000 ml of ethanol was added, and the mixture was stirred and extracted in a dark place at room temperature for 7 days. This was centrifuged and filtered under pressure to obtain an extract. This extract was freeze-dried to obtain an extract. Production example 13 Scientific name: Kyllinga monocephala Indonesian name (Akar te
ki) 50 v / v for 100 g of crushed dried whole plant
1,000 ml of ethanol was added thereto, and the mixture was extracted with stirring at room temperature in a dark place for 7 days. This was centrifuged and filtered under pressure to obtain an extract. This extract was freeze-dried to obtain an extract. Production example 14 Scientific name: Kyllinga brevifolia Indonesian name (Akar te
ki), 50 v / v for 100 g of crushed dried whole plant
1,000 ml of ethanol was added thereto, and the mixture was extracted with stirring at room temperature in a dark place for 7 days. This was centrifuged and filtered under pressure to obtain an extract. This extract was freeze-dried to obtain an extract. Production Example 15 Scientific name: Ardisia fuliginosa To 100 g of a crushed dried whole plant under the name of Indonesia (Ajag), 1,000 ml of 50 v / v% ethanol was added, and the mixture was extracted with stirring at room temperature in a dark place for 7 days. This was centrifuged and filtered under pressure to obtain an extract. This extract was freeze-dried to obtain an extract. Production example 16 Scientific name: Entada phaseoloides Indonesian name (Tariyu)
50 v / v% for 100 g of crushed dried stem and leaf
1,000 ml of ethanol was added, and the mixture was stirred and extracted in a dark place at room temperature for 7 days. This was centrifuged and filtered under pressure to obtain an extract. This extract was freeze-dried to obtain an extract. Production Example 17 Scientific name: Alyxia reinwardti Indonesian name (Kayu polo s
ari) 50 v / v% to 100 g of crushed dried stem
1,000 ml of ethanol was added, and the mixture was stirred and extracted in a dark place at room temperature for 7 days. This was centrifuged and filtered under pressure to obtain an extract. This extract was freeze-dried to obtain an extract.

Example 1 In Vitro Measurement of the Amount of NO Produced by Macrophages 1 μg / mouse of BCG (attenuated Mycobacterium tuberculosis) known to induce macrophages was intraperitoneally administered, and 4 days later, the induced peritoneal macrophages were collected. . This was spread on a 96-well plate, incubated for 2 hours to allow macrophages to adhere to the plate, and the supernatant was removed. Each of the plant extracts obtained in the Production Examples was added to both 50 μg / ml and 10 μg / ml of lipopolysaccharide (LPS), which is an endotoxin that stimulates macrophages to produce NO.
After incubation for 4 hours, NO produced and released in the culture supernatant was colored using Griess reagent and quantified by measuring absorbance.

The above experiment was performed in two parts. The results are shown in Tables 1 and 2.

[0034]

[Table 1]

[0035]

[Table 2]

As shown in Table 1 and Table 2, scientific name: Andrographis paniculata Indonesian name (Sambi boto) Scientific name: Caesalpinia sappan Indonesian name (Kaya secang) Scientific name: Schima wallichii Indonesian name (Buah cangkok) Scientific name: Alstonia scholaris Indonesian name ( Babakan pule) Scientific name: Graptophyllum pictum Indonesian name (Daum wungu) Scientific name: Usnea spp. Indonesian name (Akar angin) Scientific name: Rheum officinale Indonesian name (Klembak) Scientific name: Sindora javanica Indonesian name (Saparantu) Scientific name: Vitex trifolia Indonesian name Scientific name: Anacardium occidentale Indonesian name (Daun jambu mete) Scientific name: Gymnopetalum leucosticum Indonesian name (Duri kemarung) Scientific name: Equisetum debile Indonesian name (Greges otot) Scientific name: Kyllinga monocephala Indonesian name (Akar teki) teki) Scientific name: Ardisia fuliginosa Indonesian name (A jag) Scientific name: Entada phaseoloides An extract of Indonesian name (Tariyu) showed a strong NO production inhibitory effect on macrophage cells. Therefore, it was shown that the plant and the extract thereof revealed by the present invention were strongly activated by BCG and suppressed NO produced from macrophages stimulated by endotoxin (LPS).

Example 2 In Vivo Measurement of the Amount of NO Produced by Macrophages NO Produced by BCG-Induced Mouse Intraperitoneal Macrophages
The amount was determined by Kondo.Y., Takano, F., Hojo H., Biochem.Pharmaco.
l, 46, 1887-1892, 1993, according to the method described below. After 5 days of ICR mouse ♂5-week-old training, 3 mice were used in each group. Each plant extract obtained in Production Example was dissolved and suspended in water, and orally administered once daily at a dose of 50 mg / kg / day. After the administration on the third day, 1 μg / mouse of BCG (attenuated Mycobacterium tuberculosis) known to induce macrophages was intraperitoneally administered, and four days later, the induced peritoneal macrophages were collected. This was spread on a 24-well plate, incubated for 2 hours to allow macrophages to adhere to the plate, and the supernatant was removed. Lipopolysaccharide (LP), an endotoxin that stimulates macrophages to produce NO
S) After adding 10 μg / ml and incubating for 24 hours, NO produced and released in the culture supernatant was colored using Griess reagent and quantified by measuring absorbance.

Table 3 shows the results of the above experiment.

[Table 3]

As shown in Table 3, Scientific name: Andrographis paniculata Indonesian name (Sambi boto) Scientific name: Alstonia scholaris Indonesian name (Babakan pule) Scientific name: Alyxia reinwardti Extract of Indonesian name (Kayu polo sari) which is strong against macrophage cells A production inhibitory effect was observed. Therefore, it was shown that the plant and the extract thereof revealed by the present invention were strongly activated by BCG and suppressed NO produced from macrophages stimulated by endotoxin (LPS).

From the results of Experimental Examples 1 and 2, it was proved that the active ingredient of the present invention had an NO production inhibiting effect.

Application Example 1 Preparation of granules: Extract powder 2 prepared according to Production Example 1
00 g to 89 g of lactose and magnesium stearate 1
g, and the mixture was tableted with a single-shot tableting machine to obtain a slug tablet having a diameter of 20 mm and a weight of about 2.3 g. The slug tablet was pulverized with an oscillator, sized and sieved to obtain granules having a particle size of 20 to 50 mesh.

Application Example 2 Preparation of tablet: 200 mg of the extract prepared in Production Example 2 was microcrystalline cellulose 20 and magnesium stearate 5 g
And a tablet having a diameter of 7 mm and a weight of 225 mg was produced by tableting with a single-shot tableting machine. One tablet of the present invention contains 200 mg of dried extract powder of Tiger root.

Application Example 3 Preparation of capsule: 500 mg of the extract prepared in Production Example 3
Was filled in a hard capsule to prepare a capsule.

Application Example 4 Production of Coffee Beverage: Produced according to a conventional method according to the formulation shown in Table 4 below.

[0045]

[Table 4] ──────────────────────────────── Component Content (%) ────────イ ン ス タ ン ト Instant coffee 1.7 Granulated sugar 5.0 Extract obtained in Production Example 4 0.05 Water (or hot water) Suitable amount ──────────────────────────────── Total 100.0 ───────────── ───────────────────

Application Example 5 Production of chewing gum: The chewing gum was produced according to a conventional method according to the formulation shown in Table 5 below.

[0047]

[Table 5] ──────────────────────────────── Component Content (%) ──────── ──────────────────────── Gum base 31.6 Granulated sugar 62.5 Glycerin 0.8 Citric acid 1.0 Sucrose palmitate 1.0 Phosphoric acid Tricalcium 2.0 Ethanol extract obtained in Production Example 5 0.1 Fragrance 1.0 ─────────────────────────────合計 Total 100.0 ────────────────────────────────

Application Example 6 Manufacture of dog food: Manufactured according to a conventional method according to the formulation shown in Table 6 below.

[0049]

[Table 6] ──────────────────────────────── Component amount (%) ──────── Flour 30.0 Cornflower 15.0 Soybean flour 15.0 Meatmeal 20.0 Sugar 5.0 Tallow fat 5. 0 Salt 1.0 Calcium phosphate 1.5 Potassium sorbate 0.3 Flavor 0.6 Propylene glycol 6.5 ──────────────────────────合計 Total 100.0 ────────────────────────────────

40 parts by weight of water was added to 100 parts by weight of the above compound, and the mixture was extruded with an extruder at 150 ° C. and a screw compression ratio of 1: 3.

[0051]

The present invention has no harmful effects on humans and animals and is extremely safe. Effectively suppresses macrophage nitric oxide production by blending it into foods and feeds, and reduces shock, hypotension, rheumatoid arthritis, ulcerative colitis, ischemic encephalopathy, tumors, insulin-dependent diabetes, etc. Useful for treatment and / or prevention.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) A61K 35/78 ADU A61K 35/78 ADU (72) Inventor Ryo Soda 4-7 Kitahama, Chuo-ku, Osaka-shi, Osaka No. 28 Sumitomo Forestry Co., Ltd. (72) Inventor Shinji Fushiya 2-8-7-601, Gobashi, Aoba-ku, Sendai City, Miyagi Prefecture (72) Inventor Fumihide Takano 8-2-231, Nagamachi, Taihaku-ku, Sendai City, Miyagi Prefecture 104 F term (reference) 4C088 AA11 AA18 AB12 AB19 AB21 AB31 AB43 AB45 AB59 AB79 AC01 AC04 AC05 AC06 AC07 AC11 BA06 BA08 BA09 BA10 CA10 MA07 NA14 ZA15 ZA36 ZA43 ZA66 ZA68 ZB08 ZB11 ZB15 ZB26 ZC20 ZC35 ZC41

Claims (3)

[Claims] [Claim 1] Scientific name: Andrographis paniculata Indonesian name (Sambi boto) Scientific name: Caesalpinia sappan Indonesian name (Kaya secang) Scientific name: Schima wallichii Indonesian name (Buah cangkok) Scientific name: Alstonia scholaris Indonesian name (Babakan pule) Indonesian name: Graptoph Name (Daum wungu) Scientific name: Usnea spp. Indonesian name (Akar angin) Scientific name: Rheum officinale Indonesian name (Klembak) Scientific name: Sindora javanica Indonesian name (Saparantu) Scientific name: Vitex trifolia Indonesian name (Legundi) Scientific name: Anacardium occidentale Indonesian name Daun jambu mete) Scientific name: Gymnopetalum leucosticum Indonesian name (Duri kemarung) Scientific name: Equisetum debile Indonesian name (Greges otot) Scientific name: Kyllinga monocephala Indonesian name (Akar teki) Scientific name: Kyllinga brevifolia Indonesian name (Akar teki sci) (Ajag) Scientific name: Entada phaseoloides Inn At least one plant selected from the group consisting of the Nesia name (Tariyu) and the scientific name: Alyxia reinwardti Indonesian name (Kayu polo sari), or extracted by a supercritical extraction method using a solvent extract of plants or carbon dioxide gas A nitric oxide production inhibitor comprising the extracted extract as an active ingredient. [Claim 2] Scientific name: Whole plant of Andrographis paniculata Indonesian name (Sambi boto) Scientific name: Caesalpinia sappan Indonesian name (Kaya secang) Kibe Scientific name: Schima wallichii Indonesian name (Buah cangkok) Real scientific name: Alstonia scholaris Indonesian name (Babakan) pule) bark Scientific name: Graptophyllum pictum Indonesian name (Daum wungu) whole plant Scientific name: Usnea spp. Indonesian name (Akar angin) whole plant Scientific name: Rheum officinale Indonesian name (Klembak) Root Scientific name: Sindora javanica Indonesian name (Saparantu) Scientific name: Vitex trifolia Indonesian name (Legundi) Leaf Scientific name: Anacardium occidentale Indonesian name (Daun jambu mete) Leaf Scientific name: Gymnopetalum leucosticum Indonesian name (Duri kemarung) Thorn Scientific name: Equisetum debile Indonesian name (Greges otot) Stem Scientific name: Kyllinga monocephala Indonesian name (Akar teki) Whole plant scientific name:
Kyllinga brevifolia Indonesian name (Akar teki) whole plant Scientific name: Ardisia fuliginosa Indonesian name (Ajag) whole plant Scientific name: Entada phaseoloides Indonesian name (Tariyu) stem and leaves and Scientific name: Alyxia reinwardti Indonesian name (Kayu polo sari) stem At least one plant selected from the group consisting of or a solvent extract of a plant or an extract extracted by a supercritical extraction method using carbon dioxide as an active ingredient. Production inhibitors. 3. The solvent is water, chloroform, methanol,
The nitric oxide production inhibitor according to claim 1 or 2, wherein the agent is at least one member selected from the group consisting of ethanol, propylene glycol and 1,3-butylene glycol.