JP2010265202A - Cell growth inhibitor - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a cell growth inhibitor originated from a plant, capable of being produced by a simple method, low in toxicity, and effective for cancer cells. <P>SOLUTION: The cell growth inhibitor includes the flower portion of elm or an extract thereof, and the leaf portion of japan or an extract thereof. The elm belongs to Ulmus hollandica, and is preferably selected from the group consisting of Vegeta, Commelin, Groenveid, Belgica, Clusius, Columella, Dodoens and Homestead. The cell growth inhibitor preferably comprises a dry powder of the flower portion of elm and a dry powder of the leaf portion of japan, or the extract of the flower portion of elm and the extract of the leaf portion of japan. <P>COPYRIGHT: (C)2011,JPO&INPIT

Description

  The present invention relates to a cell growth inhibitor comprising a component derived from an elm flower part and a component derived from a leaf part of urushi. The present invention also relates to a therapeutic agent for diseases associated with abnormal cell proliferation, comprising a component derived from an elm flower part and a component derived from a leaf part of urushi.

  Researches for developing pharmaceutical compositions using components obtained from plants growing in nature have been actively conducted. Among plants existing in nature, those that humans and non-human animals use as foods are particularly active because they are low in toxicity and easy to obtain. Up to now, various pharmaceutical compositions have been developed, and the types are extremely large.

  Elm and urushi which the inventor has paid attention to are plants that grow abundantly in nature, and are easily available. However, elm has hardly been studied as a pharmaceutical until now. Urushi is also known to have urushiol which is toxic and causes dermatitis when attached to the skin, and has a strong impression that it has a bad effect on the human body. For this reason, there were few attempts to use elm and urushi as medicines compared to other plants. Under such circumstances, it has been found that a composition combining elm and urushi has high reverse transcriptase inhibitory activity (see Patent Document 1). It has also been found that a composition combining elm and urushi is useful for the prevention or treatment of hypercholesterolemia, arteriosclerosis and liver dysfunction (see Patent Document 2). However, no investigation has been made on physiological activities other than these, and no knowledge has been obtained about the influence of the composition combining elm and urushi on cell proliferation.

  On the other hand, it is necessary to treat diseases associated with abnormal cell proliferation such as cancer. In order to treat this kind of disease, it is useful to administer a cell growth inhibitor having an action of suppressing abnormal cell growth. For this reason, various compounds and compositions having a cell growth inhibitory action have been provided so far (see, for example, Patent Documents 3 to 7).

JP-A-11-130688 (see 0047) JP 2001-354581 A (refer to 0045 to 0053) JP 10-31001 A Japanese Patent Application Laid-Open No. 11-1434 JP 2006-316001 A JP 2008-201687 A JP 2005-68038 A

  However, conventionally proposed cell growth inhibitors have problems such as complicated methods for synthesizing active compounds and methods for obtaining active compositions, and low toxicity has not been sufficiently confirmed. There are many things. For this reason, it is necessary to develop a new cell growth inhibitor. Therefore, the present inventors have intensively studied for the purpose of newly providing a cell growth inhibitor that can be produced by a simple method and has low toxicity.

  As a result of intensive studies in order to solve the above problems, the present inventors have an excellent cell growth inhibitory effect in a composition containing a component derived from an elm flower part and a component derived from a leaf part of a sea urchin As a result, the present invention has been provided.

  That is, this invention provides the cell growth inhibitor characterized by including the flower part of elm or its extract, and the leaf part of an urushi, or its extract. The elm used in the cytostatic of the present invention preferably belongs to Ulmus hollandica, and among them, wegataa (Vegeta), comelin (Commelin), Groenveid, Belgica, Crusius (Clusius) ), Columella, Dodoens and Homestead, preferably selected from the group consisting of Vegeta, Commelin and Groenveid Is preferred. The cell growth inhibitor of the present invention may comprise a powder obtained by drying elm flower parts and a powder obtained by drying leaf leaves of the elm, or an extract of elm flower parts and a leaf part of urushi. It may contain an extract.

The cell growth inhibitor of the present invention is useful, for example, as a human cell growth inhibitor, useful as a cancer cell growth inhibitor, useful as a non-retroviral cancer cell growth inhibitor, and as a breast cancer cell growth inhibitor. It is useful and useful as a colon cancer cell growth inhibitor.
Further, the present invention is that (1) an elm flower part or an extract thereof and (2) a leaf part of an urushi or an extract thereof are included in any of a plurality of compositions constituting the kit. Also provided is a featured cell growth suppression kit.

The present invention also provides a therapeutic agent for diseases associated with abnormal cell proliferation, characterized by comprising elm flower parts or extracts thereof and urushi leaf parts or extracts thereof.
Furthermore, the present invention also provides a method for treating a disease associated with abnormal cell growth, which comprises administering a composition comprising an elm flower or an extract thereof and a leaf of an urushi or an extract thereof.

  The composition of the present invention comprising an elm flower part or an extract thereof and a leaf part of an urushi or an extract thereof has a cell growth inhibitory action. Moreover, the composition of this invention can be manufactured by a simple method, and its toxicity is low.

It is a phase-contrast micrograph which shows the HeLa cell growth inhibitory effect by the cell growth inhibitor addition of this invention (refer Test example 1). It is a phase-contrast micrograph which shows the MCF-7 cell growth inhibitory effect by the cell growth inhibitor addition of this invention (refer Test example 2). It is a phase-contrast micrograph which shows the DLD-1 cell growth inhibitory effect by the cell growth inhibitor addition of this invention (refer Test example 3).

  Hereinafter, the cell growth inhibitor of the present invention will be described in detail. The description of the constituent elements described below may be made based on typical embodiments of the present invention, but the present invention is not limited to such embodiments. In the present specification, a numerical range represented by using “to” means a range including numerical values described before and after “to” as a lower limit value and an upper limit value.

The cytostatic agent of the present invention is characterized by containing elm flower parts or extracts thereof, and leaves of urushi or extracts thereof.
The type of elm used for the cell growth inhibitor of the present invention is not particularly limited. Preferred is the use of elm belonging to Ulmus hollandica. Ulmus hollandica includes a hybrid of Ulmus carpinifola and Ulmus grabra, and is relatively large. In addition, the twig surface is generally smooth, and it is common that the fruit does not bear the center of the wing piece. Ulmus Hollanda is planted in Europe as street and park trees, and there are also horticultural varieties.

  Specific plant species belonging to Ulmus Hollanda include Vegeta, Comelin, Groenveid, Belgica, Clusius, Columella, Dodoens and Dodomes. Elm such as Homestead) can be exemplified. Of these, Vegeta, Comelin, and Groenveid are particularly preferred.

  Wegetaa (Vegeta) is a very sturdy elm consisting of multiple main branches. The main branches of young trees tend to grow diagonally, but generally spread sideways as they grow. For this reason, there are many branches that are not united and extend in an arbitrary direction. The trunk has many long tears and is gray. Vegeta leaves are larger than Belgica leaves described below and flatter than Commelin leaves. The roots of the leaves are wide and the shapes are various, such as an inverted egg shape and an oval shape. The buds are generally large and shiny reddish brown. Wegetaa also includes Huntingdon elm.

  Commelin is a sturdy elm with an open canopy. Many trunks are relatively straight, with a slight spiral. Branches are sparse and thin compared to Vegeta. The color of branches is generally reddish brown. The crown is often visible from below. The leaves are generally light green and small, and the veins are generally bright. The florescence is sparse and leaves much later than Vegeta. Most of the leaves are oval and have short and sharp tips. Comelin is generally strong in the wind and slow in defoliation.

  Groenveid is a large and windy elm. The tree height is about 15 to 20 m, the apex is divided into a plurality, and the tree crown is generally in a thin and well-formed shape. The leaves are small and very dense, dark green but completely yellow in autumn. The back of the leaves has vellus hair and has a slightly dull color. Due to the slow growth, there are relatively many flowers and fruits. For this reason, there exists an advantage that the flower part required by this invention can be acquired in large quantities from one elm.

  Crusius is an elm with a well-shaped crown and strong wind resistance. Columella is an elm with a small tree. Dodoens is a sturdy elm that has dark green leaves and glossy branches. Homestead is an elm grown in the United States. Dodoens and Homestead are both similar to Wegeta.

The Ulmus hollandica used in the present invention includes a hybrid of Ulmus carpinifola and Ulmus grabra.
Ulmus carpinifola generally has a wide and elliptical tree shape with a tree height of about 25 to 30 m. The bark gets rougher with age. Many twigs are relatively thin and have no hair. In addition, depending on the species, a cork layer may be clearly formed on a shoot that grows quickly. The leaves have an inverted egg shape and are often smaller than 8 cm. There are about 12 pairs of veins on the left and right.
There are many kinds of elm belonging to Ulmus carpinifola, and examples thereof include Dampieri, Hoersholmiensis, Sarniensis, and Wuredei.

On the other hand, the Ulmus grabra is a large elm with a wide and round canopy. The roots are not well developed and the bark is gray and smooth at first, but later has shallow grooves. In general, thick brown limbs grow densely, and the skin of the crown part is pink. The leaves are 8 to 16 cm, and are more rough than Ulmus carpinifola. The petiole is short and generally has slanted leg legs covering the whole. The veins extending from side to side are 12 to 18 pairs, and the inflorescence is larger. In addition, the wing piece for flying seeds has an inverted egg shape and is actually in the center.
Specific examples of elm belonging to Ulmus grabra include Camperdownii, Exoniensis, and Horizontalis.

In the present invention, plant species and elm species belonging to Ulmus hollandica other than these specific examples can also be used.
In the present invention, plant species belonging to Ulmus hollandica and plant species obtained by crossing other elm species with other plant species can also be preferably used. Other plant species to be crossed may be other elm species or plant species other than elm. Further, the plant species to be mated may be only one type or a plurality of types. Further, a variety obtained by further mating a variety obtained by mating may be used. Thus, the cell growth inhibitor of the present invention includes all those using plant species derived from elm including Ulmus hollandica.

In the present invention, only one kind of these elm seeds may be used, or a plurality of kinds may be used in combination.
In the present invention, elm flower is used. For example, plant species derived from Ulmus hollandica are budd from winter to early spring and bloom in spring. The size of the flower bud may be small. For this reason, in this invention, it is preferable to acquire and use a flower part from the stage of a bud until a flower disperses.

Elm flower parts can be used in a state that is easy to use as a cell growth inhibitor. For example, it may be used by mixing it with a suitable component after making it into a strip or powder, or using an extract extracted with a suitable solvent.
When the flower part is cut into pieces or powder, the collected flower part may be processed as it is using a cutter, shredder, colloid mill, etc., but once dried, it is shredded and powdered. It is preferred to do so. The flower part is generally dried until the water content is less than 10% by weight, preferably less than 5% by weight, more preferably less than 3% by weight. Drying may be natural drying or mechanical drying. Moreover, when drying, it is preferable to carry out within 30 minutes after flower part collection.

  The drying temperature of the flower part is not particularly limited. Therefore, rapid heating and drying may be performed using a pressure drum heating device or electromagnetic waves. When using a pressure type drum heating device, it is preferable to heat and dry within the range of 80-140 degreeC. The drying time is usually within 2 minutes, preferably within 1 minute, more preferably about 40 to 50 seconds. Moreover, when using electromagnetic waves, such as a microwave oven, it can heat-dry for about 20 to 50 second at 600 W, for example. By heating and drying under such conditions, the activity of an undesirable enzyme contained in the flower part raw material can be suppressed or inactivated, and decomposition of the physiologically active component can be prevented to some extent.

  What was rapidly heated and dried may be used as it is, or may be used after further drying at a low temperature. When performing low temperature drying, it is preferable to carry out within the range of -5 degreeC-10 degreeC. The low temperature drying can be performed by using a hot air drying device such as a far infrared drying device, a ventilation drying device, an ice temperature drying device or the like alone or in combination. For example, after performing far infrared drying, it can be dried at ice temperature. By performing such low temperature drying, decomposition of the physiologically active ingredient can be prevented.

  The flower part can be shredded or pulverized using a device or tool suitable for the purpose. For example, it can be made into a powder of 50 to 100 μm using a colloid mill. Such shredding and pulverization may be performed before drying, after high-temperature drying, or after low-temperature drying.

  The flower part can be used as an extract as well as in the form of strips or powder in this way. The flower part to be extracted may be any of the collected flower part itself, a fragmented part of the collected flower part, a high-temperature dried part, a low-temperature dried part, and the like. In order to increase the extraction efficiency, it is preferable to fragment the flower part to some extent. The extraction solvent is preferably water or alcohol, but other extraction solvents can also be used. Examples of the alcohol solvent include methanol, ethanol, propanol, isopropanol, butanol, isobutanol, tert-butanol, pentanol, isopentanol, hexanol, isohexanol and the like. Any one of these solvents may be used or may be used in combination. For example, 30-50% ethyl alcohol or methyl alcohol aqueous solution can be used.

Extraction may be performed at room temperature or under reflux. An extraction device such as a Soxhlet extractor may also be used. Specifically, a method of extracting with a Soxhlet extractor for 30 to 60 minutes using a 50% aqueous ethanol solution at a reflux temperature can be exemplified.
The extracted extract may be used as it is as a cell growth inhibitor, but it is preferably concentrated before use to enhance the effect. The degree of concentration varies depending on the use environment. Further, the solvent may be removed until it becomes powdery, and the concentration may be adjusted by further dissolving in physiological saline after making into powder. When the cell growth inhibitor of the present invention is used as a liquid agent, the solid content concentration is preferably 1 to 20% by weight, more preferably 2 to 15% by weight, and 3 to 13% by weight. More preferably, it is still more preferably 5 to 10% by weight.

For the cell growth inhibitor of the present invention, the leaf part of an urushi or an extract thereof is used together with an elm flower part or an extract thereof.
Urushi used in the present invention can be selected from those belonging to the family Ursiaceae. Particularly preferred are leaves of plant species belonging to the genus Ursi. For example, urushi (Rhus vernicitula), yamaurushi (Rhus trichocarpa), shitaurushi (Rhus ambigna), null (Rhus javanica), yamahaze (Rhus sylvestris) can be illustrated. The urushi used is preferably a young leaf with soft leaves. In particular, those of 8 weeks or less are preferable, and those of 4 weeks or less are more preferable.

  These urushi leaf parts can be dried, powdered, chopped and extracted according to the above-mentioned method, like elm flower parts. Although the mixing ratio of the component derived from the elm flower part and the component derived from the leaf part of the urushi can be appropriately determined, it is generally within the range of 1: 0.1 to 10, preferably 1: 0.3 to 3. More preferably, it is a case where it mixes by the ratio of 1: 0.5-2.

  The composition of the present invention comprising an elm flower part or an extract thereof and a leaf part of an urushi or an extract thereof has a cell growth inhibitory action. For this reason, the proliferation of the cell can be suppressed by applying the effective amount of the cell growth inhibitor of the present invention to the cell existing outside the living body. Moreover, it can be used in order to suppress cell growth by administering an effective amount of the cell growth inhibitor of the present invention to mammals including humans.

  The cell growth inhibitor of the present invention can be used as a growth inhibitor for both viral cancer cells and non-viral cancer cells. The viral cancer cell here means a cancer cell caused by a virus, and the non-viral cancer cell means a cancer cell excluding those caused by a virus. Non-viral cancer cells are thought to be caused by various factors other than viruses. For example, ingestion of carcinogens, ultraviolet irradiation, changes in the environment surrounding cells (pH reduction, oxygen concentration reduction, etc.) It can be a factor. These factors cause, for example, DNA methylation and oxidation, and other gene mutations, resulting in non-viral cancer cells. Non-viral cancer cells include brain tumor cells, skill gastric cancer cells, lung cancer cells, skin cancer cells, pancreatic cancer cells, kidney cancer cells, colon cancer cells, liver cancer cells, breast cancer cells, cervical cancer cells, etc. Those having no viral antigen or viral gene are mentioned.

  The cell growth inhibitor of the present invention is useful, for example, as a human cell growth inhibitor, useful as a cancer cell growth inhibitor, and useful as a non-retroviral cancer cell growth inhibitor. Specifically, the cervix It is useful as a cancer cell growth inhibitor, a breast cancer cell growth inhibitor, a colon cancer cell growth inhibitor, and the like. Moreover, the cell growth inhibitor of the present invention is useful as a therapeutic agent for diseases associated with abnormal cell proliferation, for example, as a therapeutic agent for diseases associated with abnormal proliferation of human cells, and abnormal cancer cells. It is useful as a therapeutic agent for diseases associated with proliferation and is useful as a therapeutic agent for diseases associated with abnormal proliferation of non-retroviral cancer cells. Specifically, abnormalities in cervical cancer cells, breast cancer cells, and colon cancer cells It is useful as a therapeutic agent for diseases associated with excessive growth.

The cell growth inhibitor of the present invention can be used in various forms depending on the purpose of use.
For example, when the cytostatic agent of the present invention is used as a pharmaceutical, various dosage forms can be selected depending on the administration route. The cytostatic agent of the present invention can be administered orally or parenterally. For example, rectal administration, intranasal administration, buccal administration, sublingual administration, intravaginal administration, intramuscular administration, subcutaneous administration, and intravenous administration can be performed. Among them, the cell growth inhibitor of the present invention is preferably administered orally, subcutaneously or transdermally, and particularly preferably administered orally.

  As preparations suitable for oral administration, tablets, capsules, powders, fine granules, granules, solutions, syrups and the like can be mentioned. As preparations suitable for parenteral administration, injections, drops, suppositories , Inhalants, transdermal absorbents, transmucosal absorbents, patches and the like. The injection may be used for any of intravenous injection, intramuscular injection, subcutaneous injection, infusion and the like. The cell growth inhibitor of the present invention is particularly preferably an oral preparation, an injection, or a patch.

  If necessary, pharmacologically and pharmaceutically acceptable additives can be added to the cell growth inhibitor of the present invention. For example, excipients, disintegrants or disintegration aids, binders, lubricants, surfactants, coating agents, dyes, diluents, bases, solubilizers or solubilizers, isotonic agents, pH adjusters Stabilizers, propellants, adhesives, wetting agents, and the like can be used. In addition, other physiologically active ingredients (for example, button skin, seisei, carrot, litchi, wolfberry, etc.) may be added. By using these additives in appropriate combinations, the cell growth inhibitor of the present invention can have various additional functions.

  Specific examples of the excipient include starch, corn starch, sucrose, lactose, mannitol, carboxymethylcellulose, and inorganic salts.

  Examples of the disintegrant or disintegration aid include wheat starch, rice starch, corn starch, potato starch, partially pregelatinized starch, hydroxypropyl starch, sodium carboxymethyl starch, tragacanth, crystalline cellulose, methylcellulose, low-substituted hydroxypropylcellulose, Specific examples include carmellose, carmellose calcium, carmellose sodium, and croscarmellose sodium.

  Examples of the binder include agar, gelatin, wheat starch, rice starch, corn starch, potato starch, dextrin, pregelatinized starch, partially pregelatinized starch, hydroxypropyl starch, crystalline cellulose, crystalline cellulose / carmellose sodium, methylcellulose, hydroxy Propylcellulose, low-substituted hydroxypropylcellulose, hydroxypropylmethylcellulose, hydroxypropylmethylcellulose phthalate, hydroxypropylmethylcellulose acetate succinate, carmellose sodium, ethylcellulose, carboxymethylethylcellulose, hydroxyethylcellulose, pullulan, polyvinylpyrrolidone, aminoalkyl methacrylate copolymer, methacrylic Copolymers, polyvinyl acetal diethylamino acetate, polyvinyl alcohol, gum arabic, gum arabic powder, white shellac, tragacanth, purified sucrose, macrogol specific examples.

  Examples of the lubricant include wheat starch, rice starch, corn starch, sucrose fatty acid ester, stearic acid, calcium stearate, magnesium stearate, hydrous silicon dioxide, light anhydrous silicic acid, synthetic aluminum silicate, and dry aluminum hydroxide gel. Specific examples include talc, magnesium aluminate metasilicate, calcium hydrogen phosphate, anhydrous calcium hydrogen phosphate, waxes, hydrogenated oil, and polyethylene glycol.

  Examples of the surfactant include sucrose fatty acid ester, soybean lecithin, polyoxyl stearate, polyoxyethylene hydrogenated castor oil, polyoxyethylene polyoxypropylene glycol, sorbitan sesquioleate, sorbitan trioleate, sorbitan monostearate, mono Specific examples include glyceryl stearate, sorbitan monopalmitate, sorbitan monolaurate, polysorbate, sodium lauryl sulfate, and lauromacrogol.

  Moreover, the cell growth inhibitor of the present invention can be designed so that the active ingredient is sustainedly released as necessary. It can also be designed so that the active ingredient is released intensively at the required location in the body. Such sustained-release preparations and drug delivery systems can be designed and manufactured according to methods well known in the pharmaceutical industry.

  In addition, an organic or inorganic carrier can be used for the cell growth inhibitor of the present invention. Examples of such carriers include lactose, starch, vegetable and animal fats and oils. In the cell growth inhibitor of the present invention, an active substance derived from elm flower parts and urushi leaf parts can be used within a range of 0.01 to 100% by weight.

  When the cell growth inhibitor of the present invention is administered, the component derived from the elm flower part and the component derived from the leaf part of the sea urchin do not necessarily have to be administered simultaneously. For example, the component derived from the elm flower part may be administered first, and the component derived from the leaf part of the urushi may be administered later, or vice versa. Preferred is when administered simultaneously. In addition, the cell growth inhibitor of the present invention may be of a type in which a component derived from the elm flower part and a component derived from the leaf part of the urushi are contained in separate containers and mixed at the time of use. Absent. For this reason, in this invention, (1) Elm flower part or its extract and (2) Urushi leaf part or its extract are contained in either of the some composition which comprises a kit. A cell growth-suppressing pharmaceutical kit characterized by

  Component (1) and component (2) may be contained in any of a plurality of compositions constituting the pharmaceutical kit of the present invention. For example, it may be a kit comprising a composition comprising component (1) and a composition comprising component (2), or a kit comprising a plurality of compositions having different abundance ratios of component (1) and component (2). It may be. When used, these compositions may be administered as they are or in appropriate combinations.

  The administration target can be a mammal other than a human, such as a dog, or a human. The dose of the cytostatic agent of the present invention is appropriately determined according to various conditions such as the purpose of treatment or prevention, the sex, weight, age, type and degree of disease, dosage form, administration route, number of administrations, etc. of the patient. To do. For example, in the case of oral administration, it can be administered at a dose of 0.1 μg to 100 mg (dry weight of active ingredient) / kg body weight / day once to several times a day. It is not limited.

The cell growth inhibitor of the present invention is not necessarily in the form of a pharmaceutical product. For example, the cell growth inhibitor of the present invention can be used safely and effectively as a functional food or a functional beverage by including it in various foods or beverages. In particular, it can be effectively used as a food or beverage displaying that it has a cell growth inhibitory effect.
Specifically, it can be used for black tea, soft drinks, juices, candy, starchy foods, various processed foods, etc., but the form of use is not limited to these specific examples. The amount of the active substance derived from the elm flower part and the leaf part of the urushi can be set within a range of about 0.1 to 99% by weight. Moreover, you may improve food texture by adding a gelatinizer etc. as needed.

  Hereinafter, the present invention will be described in more detail with reference to Examples and Test Examples. The components, ratios, operation order, and the like shown in the following examples can be appropriately changed without departing from the gist of the present invention. Therefore, the scope of the present invention is not limited to the specific examples shown in the following examples.

Example 1
In this example, specific examples of the cell growth inhibitor (solution) of the present invention are illustrated.
Flower buds of Vegeta, Commelin and Groenveid belonging to ulmus hollandica were collected in March in Amsterdam, the Netherlands. The collected soot was pulverized using a mixer and dried at 110 to 120 ° C. until the water content became 1 to 5%. The obtained dry powder was sealed with an oxygen scavenger and stored at room temperature.

  The urushi young leaves were collected in March in Ishikawa Prefecture, pulverized using a mixer, and then dried at 110 to 120 ° C. until the water content became 1 to 5%. The obtained dry powder was sealed with an oxygen scavenger and stored at room temperature.

  Each dry powder was stored for 1 week, and then 7.5 g of elm dry powder and 7.5 g of urushi dry powder were mixed. To the mixed powder, 100 ml of 70% ethanol was added, stirred for 5 hours, and concentrated so that the solid content was 58% (weight). The concentrate was dissolved in physiological saline to obtain the solution of the present invention having concentrations of 5%, 10%, 15% and 20%.

(Example 2)
In this example, specific examples of the cell growth inhibitor (powder) of the present invention are illustrated.
The dry powder of the elm flower part obtained in Example 1, the dry powder of young urushi leaves, and starch were mixed in the amounts shown in the following table, respectively. Thereby, the mixed powder of the present invention was obtained.

(Example 3)
In this example, specific examples of the cell growth inhibitor (capsule) of the present invention are illustrated.
The capsule of the present invention was manufactured by filling the powder prepared in Example 2 into a gelatin capsule.

Example 4
In this example, specific examples of the cell growth inhibitor (injection) of the present invention are illustrated.
The dry powder of elm flower part obtained in Example 1 was mixed with 7.5 g of dry powder of young urushi leaves. The mixed powder was extracted with water at 80 ° C., and the solvent was distilled off under reduced pressure. The obtained extract was dissolved in distilled water together with sodium chloride. The dry powder, sodium chloride and distilled water were mixed in the amounts shown in the following table. The injection solution of the present invention was produced by filtering the obtained aqueous solution.

(Example 5)
In this example, specific examples of the cell growth inhibitor (functional beverage) of the present invention are illustrated.
The dry powder of elm flower part obtained in Example 1 was mixed with 7.5 g of dry powder of young urushi leaves. The mixed powder was extracted with water at 60 ° C., and the obtained extract was concentrated until the solid content weight became 10%. Then, it mixed with the following components and the ratio shown in the following table | surfaces. Thereby, the functional beverage of the present invention was obtained.

(Test Example 1)
In this test example, the HeLa cell growth inhibitory action of the cell growth inhibitor obtained in Example 1 was examined.
A HeLa cell suspension at a concentration of about 1 × 10 ⁴ cells / well was prepared using Eagle's MEM medium (containing 0.1 mM non-essential amino acid) containing 10% fetal bovine serum, and a 96-well multiwell plate (inner diameter) was prepared. 6 μm) was injected into each 100 μl. 10 μl of the solution prepared in Example 1 was added and cultured in a CO ₂ incubator (5% CO ₂ ) maintained at 37 ° C. for 2 days (the present invention). For comparison, a sample was also prepared that was cultured for 2 days under the same conditions without dropping the solution prepared in Example 1 (Comparative Example). When phase contrast micrographs of each sample after 2 days of culture were taken, it was confirmed that the sample of the present invention inhibited the growth of HeLa cells and the sample of the comparative example did not inhibit the growth of HeLa cells. (FIG. 1).
In addition, after the culture for 2 days, the medium is replaced with a phenol red-free medium, and 10 μl of a reagent that reacts with living cells and turns yellow is added in a CO ₂ incubator (5% CO ₂ ). And cultured at 37 ° C. for about 3 hours. The reagent used here was obtained by dissolving 0.65 mg of WST-1 [manufactured by Dojindo Laboratories] and 0.07 mg of 1-methoxy PMS [manufactured by Dojindo Laboratories] in 1 ml of PBS. Prepared. As a result of observing the coloration state after culturing for 3 hours, it was confirmed that the sample of the present invention was not colored yellow, and the sample of the comparative example was colored yellow. This coloration state can be quantified by measuring the absorbance at 450 nm.
In the above test procedure, 10 μl of the liquid preparation prepared in Example 1 was added, and then a sample diluted twice was prepared, and a color test was performed in the same manner as the subsequent procedure to obtain an IC50 value. The IC50 value was 0.71 to 1.11 mg / ml.

(Test Example 2)
In this test example, the MCF-7 cell growth inhibitory action of the cell growth inhibitor obtained in Example 1 was examined.
MCF-7 cell suspension at a concentration of about 1 × 10 ⁴ cells / well using Eagle's MEM medium (with 0.1 mM non-essential amino acid, 1 mM sodium pyruvate, 10 μg / mL insulin) containing 10% fetal calf serum The liquid was prepared and 100 μl was injected into a 96-well multiwell plate (inner diameter 6 mm). 10 μl of the solution prepared in Example 1 was added and cultured in a CO ₂ incubator (5% CO ₂ ) maintained at 37 ° C. for 2 days (the present invention). For comparison, a sample was also prepared that was cultured for 2 days under the same conditions without dropping the solution prepared in Example 1 (Comparative Example). When phase contrast micrographs of each sample after 2 days of culture were taken, it was confirmed that the sample of the present invention inhibited the growth of breast cancer cells, and the sample of the comparative example did not inhibit the growth of breast cancer cells. (FIG. 2).
In addition, a color test was performed in the same manner as in Test Example 1 after the above-mentioned two days of culture. As a result, it was confirmed that the sample of the present invention was not colored yellow and the sample of the comparative example was colored yellow.
Furthermore, when the IC50 value was examined in the same manner as in Test Example 1, it was 1.08 to 2.04 mg / ml.

(Test Example 3)
In this test example, the DLD-1 cell growth inhibitory action of the cell growth inhibitor obtained in Example 1 was examined.
A DLD-1 cell suspension having a concentration of about 1 × 10 ⁴ cells / well was prepared using RPMI 1640 medium containing 10% fetal calf serum, and 100 μl was injected into a 96-well multiwell plate (inner diameter 6 mm). 10 μl of the solution prepared in Example 1 was added and cultured in a CO ₂ incubator (5% CO ₂ ) maintained at 37 ° C. for 2 days (the present invention). For comparison, a sample was also prepared that was cultured for 2 days under the same conditions without dropping the solution prepared in Example 1 (Comparative Example). When phase contrast micrographs of each sample after 2 days of culture were taken, it was confirmed that the sample of the present invention inhibited the growth of colon cancer cells, and the sample of the comparative example did not inhibit the growth of colon cells. (FIG. 3).
In addition, a color test was performed in the same manner as in Test Example 1 after the above-mentioned two days of culture. As a result, it was confirmed that the sample of the present invention was not colored yellow and the sample of the comparative example was colored yellow.
Further, when the IC50 value was examined in the same manner as in Test Example 1, it was 1.33 to 2.43 mg / ml.

(Test Example 4)
The liquid preparation prepared in Example 1 was orally administered to seven adult dogs (weight 20 to 55 kg, 5 to 7 years old, Melcheese Harder, Dalmation, Great Den, and hybrid) several times a day. An adult dog administered with 3 ml of the 15% concentration solution prepared in Example 1 3 times a day for 33 to 44 days, 3 ml of the 10% to 20% concentration solution prepared in Example 1 twice a day Both adult dogs administered over 95 to 115 days and adult dogs administered with the 10% solution prepared in Example 1 3 ml twice a day for 33 days have good appetite and physical condition, depending on the administration No adverse effects were found.

(Test Example 5)
In this test example, the composition of the solution obtained in Example 1 was examined. The results are shown in Table 4 below. In Table 4,% RDA represents a food safety regulation value (allowable limit value) in humans.

  From Table 4, as long as the composition was examined, it was found that the liquid obtained in Example 1 had no adverse effects on the human body.

  The composition of the present invention comprising an elm flower part or an extract thereof and a leaf part of an urushi or an extract thereof has a cell growth inhibitory action. Moreover, the composition of this invention can be manufactured by a simple method, and its toxicity is low. Therefore, the composition of the present invention is useful as a therapeutic agent for diseases associated with abnormal cell proliferation.

Claims (15)

  A cell growth inhibitor comprising an elm flower part or an extract thereof and a leaf part of an urushi or an extract thereof.   2. The cell growth inhibitor according to claim 1, wherein the elm belongs to Ulmus hollandica.   Elm is selected from the group consisting of Vegeta, Comelin, Groenveid, Belgica, Crusius, Columella, Dodoens and Homestead. The cell growth inhibitor according to claim 2.   The cell growth inhibitor according to claim 3, wherein the elm is selected from the group consisting of Vegeta, Comelin, and Groenveid.   The cell growth inhibitor according to any one of claims 1 to 4, comprising a powder obtained by drying elm flower parts and a powder obtained by drying leaves of urushi.   The cell growth inhibitor according to any one of claims 1 to 4, comprising an extract of an elm flower part and an extract of a leaf part of a sea urchin.   The cell growth inhibitor according to any one of claims 1 to 6, wherein the cell is a human cell.   The cell growth inhibitor according to any one of claims 1 to 7, wherein the cell is a cancer cell.   The cell growth inhibitor according to claim 8, wherein the cancer cell is a non-retroviral cancer cell.   The cell growth inhibitor according to claim 8, wherein the cancer cells are breast cancer cells.   The cell growth inhibitor according to claim 8, wherein the cancer cell is a colon cancer cell.   A therapeutic agent for a disease associated with abnormal cell proliferation, comprising an elm flower part or an extract thereof and a leaf part of an urushi or an extract thereof.   The therapeutic agent for a disease associated with abnormal cell proliferation according to claim 12, wherein the cancer cell is a non-retroviral cancer cell.   The therapeutic agent for a disease associated with abnormal cell proliferation according to claim 12, wherein the cancer cell is a breast cancer cell.   The therapeutic agent for a disease associated with abnormal cell proliferation according to claim 12, wherein the cancer cell is a colon cancer cell.