JP2003061615A - Telomerase inhibitor and food composition - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a telomerase inhibitor or the like which has been eaten as a food hitherto and is safely usable as a medicine for treatment and prevention of tumors, a quasi-drug and a food from plants such as farm products which have no problem from the aspect of safety. SOLUTION: This telomerase inhibitor inhibits telomerase activity and comprises one or more kinds of plants selected from mulberry, watermelon, Actinidia polygama, chrysanthemum, onion, jujube, and peach or an extract of these plants as an active ingredient.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention is derived from a natural product,
The present invention relates to a telomerase inhibitor and a food composition used for treating or preventing a disease associated with telomerase activity.

[0002]

2. Description of the Related Art Telomere, which is the terminal structure of chromosome in eukaryote, plays an important role in stabilizing the chromosome structure, and telomere DNA, which is a DNA component thereof, has many short base sequences. It has a characteristic structure that is repeated. In the case of human, this repeating unit is the 6-nucleotide sequence of TTAGGG, and the length of telomeric DNA is about 10 kb in human somatic cells. Telomere DNA is shortened for each replication due to the end replication problem in the replication of linear DNA, and if the shortening exceeds the allowable limit, destabilization of the chromosome occurs and cells die (protein nucleic acid enzyme, 42, 1351-. 1374 pages,
1997).

On the other hand, cancer cells are abnormal cells characterized by unregulated and unlimited proliferation. Cancer cells need to compensate for the above-mentioned telomere shortening in order to allow unlimited proliferation (immortalization), and the enzyme telomerase plays an important role in the compensation.

This telomerase is an RNA-dependent DNA polymerase that extends telomeric DNA using RNA as a template, and is composed of template RNA (TR) and catalytic subunit protein (TERT). Telomerase activity has been found in most of the cancer cells and immortalized cells examined (Science, 266, 2011-201.
On the other hand, no activity is observed in normal somatic cells except for some lymphocytes and the like. Therefore, telomerase is regarded as a promising target for antitumor agents, and telomerase inhibitors do not affect normal cells and are expected to selectively act only on tumor cells. In fact, it has been reported that the expression of mutant TERT inhibits the telomerase activity of cancer cells, leading to the death of cancer cells (N
atureMedicine, 5 volumes, 1164-117
Page 0, 1999; Genes & Development.
nt, vol. 13, pp. 2388-2399, 1999),
It has also been reported that peptide nucleic acid-based telomerase inhibitors bring about telomere shortening and growth inhibition (Pr.
ocedings of the National
Academy of Sciences of t
he Unites States of America
a, 96, 14276-14281, 1999; O.
ncogene, 18: 6191-6200, 19
1999), it was further found that the expression of antisense to TR in cancer cells enhances the sensitivity to the anticancer drug cisplatin (Oncog).
ene, vol. 16, pp. 2243-2248, 1998.
Year).

Development of telomerase inhibitors has been attempted so far, and synthetic ribozyme (Antis) has been tried.
sense & Nucleic Acid Drug
Development, Volume 8, 309-317, 1
998), modified oligonucleotides (Proceed
ings of the National Acad
emy of Sciences of the Un
ites States of America, 95
Vol., 11549-11554, 1998), Peptide Nucleic Acid Derivatives (Nature Biotechnolo).
gy, 14: 615-619, 1996), antisense DNA (FASEB Journal, 12).
Vol. 801-811, 1998; Oncogen.
e, 16: 3323-3330, 1998), triazine derivatives (JP-A-11-60573), pyridine derivatives (JP-A-11-49676; JP-A-11-49).
678; JP-A-11-049777), thiazoline derivatives (JP-A-2001-72592), secondary metabolites of microorganisms (JP-A-2001-81029; Biochemis).
try, 39, 5995-6002, 2000,
Journal of the American Ch
electronic Society, Volume 123, 1262-
1263, 2001; Bioscience, B.
iotechnology, and Biochem
istry, 65, 651-653, 2001;
Oncology Research, 10 volumes, 449
~ 453, 1998), Marine microalgae extract (R
essearch Communications in
Molecular Pathology and
Pharmacology, Volume 99, 259-265
P., 1998; Cytotechnology, 33.
Vol. 221-227, 2000), Green tea catechins (Biochemical and Biophysi).
cal Research Communicatio
ns, 249, 391-396, 1998).

However, at present, pharmaceuticals and functional foods whose mechanism of action is telomerase inhibition have not been put into practical use, and most of the above telomerase inhibitors are available as pharmaceuticals. However, since it is a chemically synthesized product, there is a problem in safety when it is used as a food, and it must be said that its application as a food for carcinostatic is difficult.

The present invention solves the above problems,
A telomerase inhibitor and a food that can be safely used as a drug, quasi drug, and food that can be used to treat or prevent tumors from plants such as agricultural products that have been ingested as foods and have no safety problems. A composition is provided.

[0008]

The gist of the present invention will be described.

It inhibits the activity of telomerase, and includes mulberry, watermelon, caterpillar, chrysanthemum, onion, jujube,
The present invention relates to a telomerase inhibitor, which comprises one or more plants selected from peach or an extract of these plants as an active ingredient.

In addition, it exerts an antitumor effect by inhibiting telomerase activity.
1 selected from Matatabi, chrysanthemum, onion, jujube, peach
The present invention relates to a telomerase inhibitor, which comprises one or more plants or an extract of these plants as an active ingredient.

[0011] In addition, one or more plants selected from mulberry, watermelon, matabi, chrysanthemum, onion, jujube and peach, which inhibit telomerase activity, or an extract of these plants is used as an active ingredient. The present invention relates to a food composition characterized by:

[0012] Further, it exerts an antitumor effect by inhibiting telomerase activity.
1 selected from Matatabi, chrysanthemum, onion, jujube, peach
The present invention relates to a food composition comprising at least one plant or an extract of these plants as an active ingredient.

[0013] In addition, the telomerase activity is inhibited, and an organic solvent extract or water extract of at least one plant selected from mulberry, watermelon, matabi, chrysanthemum, onion, jujube, and peach is used as an active ingredient. The present invention relates to a telomerase inhibitor characterized in that

Further, a substance which inhibits the telomerase activity and which is obtained by concentrating an organic solvent extract or a water extract of one or more plants selected from mulberry, watermelon, matabi, chrysanthemum, onion, jujube and peach is used. The present invention relates to a telomerase inhibitor characterized by being an active ingredient.

[0015]

The present invention has been achieved by confirming the effect as a result of repeated experiments, and has been achieved by mulberry, watermelon, matabi, chrysanthemum, onion, jujube and peach, or telomerase of these extracts. As a result of diligent examination of the inhibitory action, it was newly found that these all have telomerase inhibitory activity and therefore can be used as components of antitumor agents and antitumor foods.

Mulberry, watermelon, matabi, chrysanthemum, onion, jujube and peach are commonly used as foods, but it has never been known that they have a telomerase inhibitory action.

Since the present invention is constituted as described above, it is a safe telomerase inhibitor and a food composition exhibiting a telomerase inhibitory action.

[0018]

BEST MODE FOR CARRYING OUT THE INVENTION Embodiments of the present invention will be described below.

In this embodiment, mulberry, watermelon, matatabi, chrysanthemum,
One or more plants selected from onions, jujubes and peaches,
Alternatively, the present invention relates to a telomerase inhibitor, a telomerase-inhibiting antitumor agent, and a telomerase-inhibiting food containing an extract of these plants as an active ingredient and having a property of inhibiting telomerase activity.

[0020] Mulberry is a plant of the Moraceae family and belongs to the Yamaguchi (Morus bombycis Ko).
idz. ) And Magui (M. alba L.) are known. Mulberry fruit is used as a crude drug and is well known as a raw material for raw food and mulberry wine.
Mulberry leaves have long been used as mulberry tea.

Watermelon (Citrullus) belongs to the family Cucurbitaceae and is widely cultivated for food. The fruit is mainly used for raw food.

Matabi (Actinidia poly)
gama Maxim. ) Is the Actinidia family (Actini)
It is a shrub with climbing ability that belongs to diaceae). The fruit of Matatabi is salted and eaten, and it is also used as a crude drug.

Chrysanthemum is a plant of the family Asteraceae (Compositae) and is widely cultivated for horticulture, while its head flowers are dried and used as edible chrysanthemum.

Onion (Allium cepa
L. ) Belongs to the family Lilyaceae, and has been widely cultivated in Japan since the Meiji era for food. The bulb is commonly used as a raw material for cooking vegetables and foods.

Zizypus is a deciduous small tree belonging to the family Rhamnaceae, and its fruit is used as a raw material for producing confectionery.

Peach (Prunus persica) belongs to the family Rosaceae, flowers are widely used for ornamental purposes, and fruits are widely used for food including raw food.

These plants can be whole or leaf, stem,
Some flowers, fruits, roots, seeds, etc. can also be used as telomerase inhibitors. You can use these as they are,
It can also be used as a dried product. Also, these can be used as a crushed product or an extract.

Further, these plants are preferably used as an extract in consideration of handling property and the like.

As the extraction method, a method using water or a method using an organic solvent can be used. The organic solvent used is not particularly limited, and examples thereof include alcohols such as methyl alcohol, ethyl alcohol and butyl alcohol, esters such as ethyl acetate, hydrocarbons such as hexane and petroleum ether, and halogenated hydrocarbons. Kinds and the like can be used. Further, these solvents can be used alone or as a mixture of two or more kinds, and a method of sequentially extracting with a plurality of solvents is also effective.

This extract can be used as a liquid as it is, but may be used in combination with a treatment method generally used for preparation of the extract, such as concentration, concentration to dryness and freeze-drying. Also,
The plant and its extract can be used not only alone but also as a mixture of two or more kinds.

The plant and the extract thereof thus obtained exert an excellent telomerase inhibitory action.

Since these plants have been widely ingested as foods, they are highly safe.

Therefore, it can be applied not only to drugs and quasi-drugs but also to foods for the purpose of treating and preventing tumors associated with telomerase.

The use form of the plant or its extract is not particularly limited as long as it can be expected to have a telomerase inhibitory action. Examples of the medicine include liquid preparations such as injectable solutions containing these plants and their extracts as active ingredients, or solid preparations such as tablets and coatings,
It is manufactured by a conventional method alone or mixed with an excipient. The administration method can be either oral or parenteral.

Examples of quasi-drugs include bathing agents and creams containing these plants and their extracts as active ingredients, but are not particularly limited as long as they have a shape that can be used as quasi-drugs.

As foods, these plants and their extracts can be eaten directly or after cooking, but they can also be processed into various foods containing these plants and their extracts as active ingredients. is there. For example, it can be variously used as a processing component such as beverages such as juice, confectioneries such as rice crackers, seasonings, breads and noodles.

Since this example is constituted as described above, it is a safe telomerase inhibitor and the like that can be used for the treatment and prevention of diseases such as malignant tumors where the telomerase inhibitory action is effective.

Experimental results confirming the effects of this embodiment will be described below.

Experimental Example 1 (Production of Extract from Mulberry Leaf) Mulberry leaf was shredded and then refluxed with methanol for 3 hours according to a conventional method. After removing the residue by filtration, the methanol layer was concentrated to dryness. The residue is ethyl acetate / water = 1: 1 (volume ratio)
The ethyl acetate layer was concentrated to dryness, and methanol-
0.045 g of an ethyl acetate extract was obtained.

Experimental Example 2 (Production of Extract from Watermelon Fruit) Watermelon fruit was shredded and then refluxed with methanol for 3 hours according to a conventional method. After removing the residue by filtration, the methanol layer was concentrated to dryness. The residue was fractionated with ether / water = 1: 1 (volume ratio), and the ether layer was concentrated to dryness to obtain 0.446 g of a methanol-ether extract. Further, the same volume of ethyl acetate was added to the aqueous layer for further fractionation, and the ethyl acetate layer was separated and concentrated to dryness to obtain 0.393 g of a methanol-ethyl acetate extract.

Experimental Example 3 (Production of Extract from Matatabi Fruit) The Matatabi fruit was crushed and then refluxed with methanol for 3 hours by a conventional method. After removing the residue by filtration, the methanol layer was concentrated to dryness. The residue was fractionated with ether / water = 1: 1 (volume ratio), and the ether layer was concentrated to dryness to obtain 2.691 g of a methanol-ether extract.

Experimental Example 4 (Production of Extract from Edible Chrysanthemum) After freeze-dried preparation of edible chrysanthemum was pulverized, it was refluxed with methanol for 3 hours according to a conventional method. After removing the residue by filtration, the methanol layer was concentrated to dryness. The residue is ethyl acetate / water =
Fractionation was performed at a volume ratio of 1: 1 and the ethyl acetate layer was concentrated to dryness to obtain 0.730 g of a methanol-ethyl acetate extract. The same volume of butyl alcohol was added to the aqueous layer for further fractionation. The butyl alcohol layer was separated and concentrated to dryness to give methanol-butyl alcohol extract 1.45.
4 g was obtained.

Experimental Example 5 (Production of Extract from Onion Bulb) A freeze-dried preparation of onion bulb was pulverized and then refluxed with methanol for 3 hours according to a conventional method. After removing the residue by filtration, the methanol layer was concentrated to dryness. The residue is ether /
Fractionation was performed with water = 1: 1 (volume ratio), and the ether layer was concentrated to dryness to obtain 0.163 g of a methanol-ether extract. Further, the same volume of ethyl acetate was added to the aqueous layer for further fractionation, and the ethyl acetate layer was separated and concentrated to dryness to obtain 0.049 g of a methanol-ethyl acetate extract. After adding the same volume of butyl alcohol to the aqueous layer for fractionation,
The butyl alcohol layer was concentrated to dryness to obtain 1.260 g of a methanol-butyl alcohol extract.

Experimental Example 6 (Production of Extract from Jujube Fruit) Jujube fruit was crushed and then refluxed with methanol for 3 hours according to a conventional method. After removing the residue by filtration, the methanol layer was concentrated to dryness. The residue was fractionated with ether / water = 1: 1 (volume ratio), and the ether layer was concentrated to dryness to obtain 1.394 g of a methanol-ether extract. Further, the same volume of ethyl acetate was added to the aqueous layer for further fractionation, and the ethyl acetate layer was separated and concentrated to dryness to obtain 0.336 g of a methanol-ethyl acetate extract. The aqueous layer was further fractionated by adding the same volume of butyl alcohol, and the butyl alcohol layer was concentrated to dryness to obtain 3.109 g of a methanol-butyl alcohol extract. The remaining aqueous layer was freeze-dried to obtain 91.149 g of a methanol-water extract.

Experimental Example 7 (Production of Extract from Peach Fruit) A freeze-dried preparation of peach fruit was pulverized and then refluxed with methanol for 3 hours according to a conventional method. After removing the methanol layer, the residue was refluxed with water for 3 hours. After refluxing, the residue was filtered off, and the remaining aqueous layer was freeze-dried to obtain 6.747 g of a water extract.

Experimental Example 8 (Test for Telomerase Inhibitory Activity) The telomerase activity and its inhibitory activity by plant extracts were determined by Ishikawa et al. (Oncogene, vol. 13,
2265-2274, 1996) based on the stretch PCR method proposed (Toyobo Co., Te
loChaser Telomerase Assay
Kit). Cell extract containing telomerase was prepared from human tumor cell line U937 (JCRB9021). For the preparation of the cell extract, the above measurement kit was used as in the case of measuring the telomerase activity. The test extract was added at a predetermined concentration during the telomerase reaction, and the relative value of the telomerase activity when the test extract was added was calculated assuming that the telomerase activity without addition was 100%. The results are shown in Table 1 below. It is clear from the table that these plant extracts show excellent telomerase inhibitory action.

─────────────────────────────────────────────────── ─── Continuation of front page (51) Int.Cl. ⁷ Identification code FI theme code (reference) A61K 35/78 A61K 35/78 TV A61P 35/00 A61P 35/00 43/00 111 43/00 111 C12N 9/99 C12N 9/99 (72) Inventor Masatoshi Kusunoki 2-25 Shinei-cho, Kamo-shi, Niigata Niigata Agricultural Research Institute Food Research Center F-term (reference) 4B018 MD52 MD53 MD61 ME08 MF01 4C088 AB12 AB19 AB26 AB34 AB51 AB87 AC03 AC04 AC05 CA06 CA07 NA14 ZB26 ZC20

Claims (6)

[Claims] 1. An active ingredient which inhibits telomerase activity, and which comprises at least one plant selected from mulberry, watermelon, matabi, chrysanthemum, onion, jujube and peach, or an extract of these plants. A telomerase inhibitor characterized by: 2. One or more plants selected from mulberry, watermelon, matabi, chrysanthemum, onion, jujube, and peach, which exhibit an antitumor effect by inhibiting telomerase activity, or a combination of these plants. A telomerase inhibitor comprising an extract as an active ingredient. 3. An active ingredient which inhibits telomerase activity, and comprises one or more plants selected from mulberry, watermelon, matabi, chrysanthemum, onion, jujube and peach, or an extract of these plants. A food composition characterized by: 4. An antitumor effect is exhibited by inhibiting telomerase activity, which comprises one or more plants selected from mulberry, watermelon, matabi, chrysanthemum, onion, jujube and peach, or a plant of these plants. A food composition comprising an extract as an active ingredient. 5. An active ingredient comprising an organic solvent extract or water extract of one or more plants selected from mulberry, watermelon, matabi, chrysanthemum, onion, jujube, and peach, which inhibits telomerase activity. A telomerase inhibitor characterized in that 6. A substance which inhibits telomerase activity and which is obtained by concentrating an organic solvent extract or water extract of one or more plants selected from mulberry, watermelon, matabi, chrysanthemum, onion, jujube, and peach. A telomerase inhibitor, which is an active ingredient.