KR20090038699A - A composition for preventing or treating cancer comprising stem extract of rosa rugosa - Google Patents

Abstract

A pharmaceutical composition with Rosa rugosa stem extracts is provided to treat and prevent hormone receptor-mediated cancer such as prostate cancer by inhibiting activity of histone acetyltransferase(HAT). A pharmaceutical composition for treatment and prevention of hormone receptor-mediated cancer comprises Rosa rugosa stem extracts as active ingredients. The Rosa rugosa stem extracts are extracted by water, C1-C6 low alcohol or a mixed solvent thereof. The hormone receptor-mediated cancer represents prostate cancer. The pharmaceutical composition for treatment and prevention of hormone receptor-mediated cancer additionally contains pharmaceutically allowable carriers and Flutamide.

Description

A composition for preventing or treating cancer comprising stem extract of Rosa rugosa}

The present invention relates to a composition for preventing or treating cancer, which comprises a glycolytic stem extract. More specifically, the present invention relates to novel uses of glycolytic stem extracts having histone acetyltransferase (HAT) inhibitory ability as natural materials for androgen receptor mediated cancers, particularly anticancer drugs and health supplements for prostate cancer.

Prostate cancer is the leading cause of death among adult males in the United States, and the recent incidence has increased rapidly in Korea due to changes in diet and lifestyle. Androgen receptors, which are essential for the development and progression of prostate cancer, belong to the steroid hormone receptor superfamily. Androgen, a male hormone, is essential for the development and maintenance of the reproductive system and the growth of the prostate gland. Modulates the activity of receptors (Park, SK et al . Prostate 66 (12): 1285-91 (2006); Taplin ME et al . J Cell Biochem . 15; 91 (3): 483-90, 2004; Chen, L. et al . Mol Cancer Ther . 4 (9): 1311-9, 2005; Yoon, HG et al . Mol Endocrinol . 20 (5): 1048-60, 2006).

Histones are tightly bound to DNA with genetic information to form nucleosomes in the nucleus, and post-translational modifications of these DNA-binding proteins can regulate gene expression and signaling. The acetylation of histones during this transformation is largely regulated by two enzymes: histone acetyltransferase (HAT) and histone deacetylase (HDAC). It can also regulate the activity of nonhistone proteins. HAT proteins increase the acetylation of hormone receptors, and acetylated hormone receptors overexpress each target protein, causing cancer growth and development. As a result, inhibition of hyperacetylation in hormone receptor-related cancers has been recognized as a molecular target for the search for new cancer inhibitors. In particular, hyperacetylation of androgen receptors and histones is associated with the development and proliferation of androgen receptor-independent prostate cancer. Related to cancer cell lines in In vitro experiments have reported apoptosis suppression mechanisms (Kang, J. et. al . Biochem Pharmacol . 69 (8): 1205-13, 2005; Stimson, L. et al . Mol Cancer Ther . 4 (10): 1521-32, 2005; Balasubramanya, K. et al . J Biol Chem . 279 (49): 51163-71, 2004; Druesne, N. et al . Carcinogenesis 25 (7): 1227-36, 2004; Fu, M. et al . Biochem Pharmacol . 68 (6): 1199-208, 2004).

In order to control such acetylation, inhibitors of HDAC activity have been released, mainly using HDAC protein as a target protein for cancer treatment. Relatively limited HAT inhibitors compared to these various HDAC inhibitors have been published. Inhibitors studied to date include synthetic peptide-CoA conjugates, isothiazolone, polyprenylated benzophenone, curcumin, and anacardic acid (Kang) , J. et al . Biochem Pharmacol . 69 (8): 1205-13, 2005; Inche, AG Drug Discov Today . 11 (3-4): 97-109, 2006; Lau, OD et al . Mol Cell . 5 (3): 589-95, 2000; Debes, JD et al . Cancer Res . 63 (22): 7638-40, 2003; Sun, Y. et al , FEBS Lett . 580 (18): 4353-6, 2006). Recently, studies on plant-derived inhibitors that are easier to secure safety than synthetic compounds such as peptide-CoA conjugate and isothiazolone have been actively conducted. Curcuminn in turmeric mentioned above, anacardic acid from cashew nuts, and Garcinia benzophenone in indica ) is an example of a plant-derived HAT inhibitor. The securing of such HAT inhibitory activity-containing plant extracts is constantly needed for the development of new anticancer drugs.

Pervasive ( Rosa rugosa ) is a genus of rose family, distributed in Northeast Asia, and native to the sandy beaches of the sea. It uses medicinal fruits and roots for medicinal purposes, and has been reported on the functionality of flowers, but very few reports or use of stems is known. Stem sites contain catechins, gallic acid, methyl gallate, quercetin, hyperosides, etc. (Y. Hashidoki, Phytochemistry . 43: 535-549, 1996; BM Schmidt et al . Cancer lett . 231; 240-246, 2006), which have not been reported as inhibitors of HAT activity for the prevention of hormone receptor-related cancers, including prostate cancer.

An object of the present invention is to provide a prophylactic or therapeutic use of cancer, in particular hormone receptor mediated cancer, such as prostate cancer, by examining the inhibitory ability of HAT activity of glycolysis stem extract.

Another object of the present invention is to provide a pharmaceutical preparation and dietary supplement for cancer prevention, improvement or treatment comprising the corresponding stem extract as an active ingredient.

In order to achieve the above object, the present invention provides a composition for the prevention or treatment of cancer comprising the corresponding stem extract as an active ingredient.

The composition according to the invention is preferably used as a pharmaceutical preparation for the prevention or treatment of hormone receptor mediated cancer, in particular prostate cancer, comprising an effective amount of a glycolytic stem extract. The pharmaceutical formulation may further comprise a pharmaceutically acceptable carrier.

In addition, the composition according to the invention is preferably used as a dietary supplement for cancer prevention or improvement comprising an effective amount of the corresponding stem extract. The health supplement is preferably used as a food or food additive, beverage or beverage additive.

Cancer preventive or therapeutic composition comprising the corresponding stem extract of the present invention as an active ingredient is excellent in inhibiting the activity of histone acetyltransferase to prevent, improve or prevent cancer, in particular hormone receptor mediated cancer, such as prostate cancer or It is excellent for treatment.

Hereinafter, the present invention will be described in detail.

The present invention is a nuclear protein (nuclear) extracted from the extract of the plant and medicinal plants of about 500 species from the LNCaP cell line which is a human prostate cancer cell line extract derived from the plant having an inhibitory activity of HAT involved in the growth and development of hormone-mediated cancer extract), and P300 and CREB-binding protein (CBP) were identified as inhibitory activity against specific HAT protein. Using the LNCaP cell line in vitro The present invention was completed by obtaining an extract of a glycolytic stem exhibiting anticancer activity by confirming molecular mechanisms of growth inhibition and transcriptional activity inhibition and mRNA expression of androgen receptor regulatory genes in cancer cells in the stomach.

Therefore, the present invention is characterized by providing a composition for preventing or treating cancer, which comprises a glycolytic stem extract, which is an inhibitor of HAT activity, as an active ingredient.

In the inhibitor of HAT activity, the mechanism of action of HAT which is closely related to prostate cancer and the treatment of anticancer or anti-inflammatory or acquired immunodeficiency syndrome by them are well known in the art (Stimson, L. et. al . Mol Cancer Ther . 4 (10): 1521-32, 2005; Balasubramanyam, K. et al . J Biol Chem . 279 (49): 51163-71, 2004; Balasubramanyam, K. et al . J Biol Chem . 279 (32): 33716-26, 2004; Adcock, I. et al. Curr Opin Immunol . 19: 1-7) It is apparent that glycolytic stem extracts that inhibit HAT activity are useful as prophylactic and / or therapeutic agents for hormone receptor mediated cancer, particularly prostate cancer, or inflammation or AIDS.

The composition for preventing or treating cancer according to the present invention may be prepared as a pharmaceutical preparation for preventing or treating cancer, in particular hormone receptor mediated cancer, for example prostate cancer.

The pharmaceutical formulation is characterized by having a very high safety and no side effects compared to conventional organic synthetic drugs by using a natural material separated from the corresponding stem.

Separation method of the glycolylated stem extract of the present invention is not particularly limited, but extracted with water or lower alcohol of 1 to 6 carbon atoms or a mixed solvent thereof at 90 to 100 ℃ concentrated under reduced pressure or lower alcohol of 1 to 6 carbon atoms, In particular, the methanol extract may be further fractionated with various organic solvents such as hexane, chloroform, ethyl acetate or butanol, and may be further purified by various chromatography after the organic solvent fractionation.

The applicable stem extract of the present invention shall include any one of all extracts, fractions and purified products obtained in each step of extraction, fractionation and purification, dilutions or concentrates thereof or dried products thereof. Preferably, hot water or methanol extracts of glycolysis stems, more preferably butanol fractions of methanol extracts are used.

In addition, the composition for preventing or treating cancer with the effective amount of the glycolysis stem extract of the present invention is flutamide (Slovin SF Nat Clin Pract), which is known as a prostate cancer treatment agent. Oncol . 2007 Sep; 4 (9): 551-4; Bennett CL et al . Prostate Cancer Prostatic Dis . 1999 Jan; 2 (1): 4-8).

The flutamide is preferably included in a final concentration of 10-30μM for the composition of the present invention. More preferably, it is contained in 20 micrometers. When flutamide is used in combination with the corresponding stem extract, no synergistic effect on the inhibition of HAT is observed at concentrations below 10 μM, and the synergistic effect on the inhibition of HAT does not increase more than 30 μM. Considering side effects such as bowel dysfunction, it is good to include 10 ~ 30μM.

The composition for preventing or treating cancer of the present invention may be formulated in the form of granules, tablets, capsules or drinks by conventional methods known in the art. Moreover, in order to make storage and handling easy, you may add the carrier used for normal formulation, such as dextrin and cyclodextrin, and other arbitrary preparations. In addition, auxiliary raw materials or additives conventionally added to foods or drugs are not particularly limited, for example, glucose, fructose, sucrose, maltose, sorbitol, stevioside, loxoside, corn syrup, lactose, citric acid, tartaric acid, malic acid , Succinic acid, lactic acid, L-ascorbic acid, d1-α-tocopherol, sodium elisolvinate, glycerin, propylene glycol, glycerin fatty acid ester, polyglycerin fatty acid ester, sucrose fatty acid ester, sorbitan fatty acid ester, gum arabic , Casein, gelatin, pectin, agar, vitamin Bs, nicotinic acid amide, calcium panthenate, amino acids, calcium salts, pigments, flavors, preservatives and the like.

In addition, the present invention provides a health supplement for the prevention or improvement of cancer, in particular hormone receptor-mediated cancer, for example, prostate cancer, comprising a glycolytic stem extract as an active ingredient.

A dietary supplement is a food prepared by adding the relevant stem extract to food materials such as beverages, teas, spices, gums, confectionery, or by encapsulating, powdering, and suspension. This means, unlike general medicine, there is no side effect that can occur when taking a long-term use of the drug as a food raw material. Since the dietary supplement of the present invention thus obtained can be consumed on a daily basis, a high anticancer and cancer prevention effect can be expected, which is very useful.

In the dietary supplement of the present invention, the addition amount of the glycolytic stem extract cannot be uniformly defined depending on the kind of the dietary supplement targeted, but may be added within a range that does not impair the original taste of the food, 0.01 to 50% by weight, preferably 0.1 to 20% by weight. In the case of food in the form of granules, tablets or capsules, it is usually added in the range of 0.1 to 100% by weight, preferably 5 to 100% by weight.

In addition, in the case of a pharmaceutical formulation containing an effective amount of the glycolytic stem extract of the present invention, it is appropriate to administer so that the daily intake of the adult is 1 to 3,000 mg. In addition, the dosage can be appropriately increased or decreased depending on age, symptoms, and the like.

Next will be described in more detail with reference to the embodiment of the present invention configured as described above. These examples are only for illustrating the present invention in detail, it will be apparent to those skilled in the art that the scope of the present invention is not limited to these examples.

Example 1 Preparation of Glycogenated Stem Hot Water and Methanol Extract

In the case of hot water extraction, 1.2 kg of the corresponding stems were blanched at 100 ° C. for 5 minutes, and reflux extraction was performed by adding 5 times the amount of distilled water of the sample. After standing still, this process was repeated three times, and the extract was concentrated under reduced pressure and dried to obtain a hydrolyzed stem hydrothermal and methanol extract.

Example 2 Investigation of HAT Inhibition Activity of Glycrum Stem Extract

LNCaP cells (American Cell Type Collection (ATCC, VA, USA)), human prostate cancer cell lines, were grown to complete confluence on 150 mm plates and collected, and then pelleted by centrifugation at 1,000 rpm for 3 minutes. Was collected. 20 ml of cold buffer A (cold buffer A, 10mM pH 7.9 Hepes, 1.5mM MgCl ₂ , 10mM KCl, 0.5mM DTT, 0.2mM PMSF) was added to the collected pellets, and the mixture was left for 10 minutes on ice. After centrifugation for 5 minutes, the pellets were collected again, homogenized by adding 6 mL of the buffer solution A as described above. Centrifuge the prepared cell homogenate at 4 ° C., 12,500 rpm for 30 minutes and collect pellets for buffer C (20 mM pH 7.9 Hepes, 25% Glycerol, 0.42 M NaCl, 1.5 mM MgCl ₂ , 0.2 mM EDTA, 0.5 mM DTT, 0.2mM PMSF) 4mL was added, resuspended, and left to stand on ice for 30 minutes, followed by centrifugation at 4, 12,500 rpm for 30 minutes. Finally, the supernatant obtained by centrifuging the supernatant at 4 ° C. and 30,000 rpm for 1 hour was collected and used for the experiment.

In order to measure HAT inhibitory activity, 5 μg of the nuclear extract prepared above and hot water or methanol extract of glycolytic stem were added to measure activity at 440 nm of absorbance using a colorimetric kit for measuring HAT activity (Biovision, CA, USA). Here, HAT activity inhibition capacity (%) was converted according to the following formula.

% Inhibition of HAT activity = [1- (As-Ab / Ac-Ab)] × 100

Ac: absorbance at distilled water treatment

Ab: absorbance of blank

As: absorbance of sample treatment

As a result, the hydrothermal extract of the glycolytic stem showed less than 20% inhibitory effect at a concentration of 100 µg / mL, while the methanol extract showed an inhibitory effect of nearly 40%, and thus the methanol extract was used in the experiment. Shown).

When the methanol extract of the glycolytic stem was treated at concentrations of 0, 50, 100, and 200 µg / mL, as shown in FIG. 1, 25-35% inhibitory activity was confirmed.

<Comparative Example 1> Comparison of HAT Inhibitory Activity by Related Sites

In order to compare the inhibitory activity of HAT activity of glycolysis root and fruit parts and stem extracts, which have known pharmacological effects to date, the HAT inhibitory activity was examined according to the method described in Example 2 with the methanol extract of each site at the final concentration of 100 µg / mL. It was.

As shown in FIG. 2, the stem extract showed 34.4% inhibitory activity, while the root and fruit extracts were found to have no HAT inhibitory activity.

Example 3 Investigation of Solvent Fraction and HAT Inhibitory Activity of Methanol Extract of Glycated Stem

Solvent fractionation was carried out on the methanol extract of the glycolysis stem confirmed the HAT inhibitory activity was traced the change of the HAT inhibitory activity.

To this end, finely pulverized the stem of the glycolysis, and then lanching for 5 minutes at 100 ℃ to inactivate enzymes and pulverize the biological cells, and then extracted by standing for 5 hours with 95% methanol of the sample at room temperature three times After repeated, the extract was concentrated under reduced pressure. 1 L of hexane, chloroform and butanol were added to the concentrate, and the solvent was exchanged three times for 36 hours in each solvent. The residue was partitioned with a funnel, and the residue was dissolved in water. Concentrated and dried to obtain an extract (see Figure 3).

As shown in Figure 4, it was confirmed that the butanol fraction (hereinafter referred to as "RMB-1") shows excellent HAT inhibitory activity. Therefore, in the following examples, butanol fraction of the methanol extract of the glycolysis stem was used for the experiment.

Example 4 Investigation of Enzyme-Specific HAT Inhibitory Activity by Immunoprecipitation (IP, Immunoprecipitation) of Glycylated Stem Butanol Fraction

In order to measure the activity of enzyme-specific HAT proteins such as P300 and CBP and examine the inhibition specificity, 50 µg of LNCaP nuclear extract prepared in the same manner as in Example 2 was previously applied at 4 ° C. for 2 hours. After pre-clearing, protein A / G agarose bead (10 μl, Santa Cruz, CA, USA) and 2 μl of p300 and CBP protein were added and washed overnight. The eluted product was eluted. product), according to the method of Example 2, the corresponding stem butanol fractions were added for each concentration (50-200 µg / mL) and subjected to HAT analysis to compare specific inhibitory activity.

As shown in Figure 5, it was confirmed that there is an inhibitory effect on CBP (50%) and P300 (70%) at the concentration of glycolytic stem butanol fraction (RMB-1) 100㎍ / mL.

CBP and p300 are directly involved in the acetylation of not only histones but also the androgen receptors in in vitro and in reported in vivo . According to the experimental results, it is thought that RMB-1 has an excellent effect of inhibiting the acetylation of the androgen receptor which is closely related to the growth of prostate cancer cells by inhibiting the HAT activity of CBP and p300.

<Example 5> Investigation of the transcriptional activity inhibitory activity of glycolytic stem butanol fraction in prostate cancer cell line

LNCaP cell line, a prostate cancer cell line, was cultured in RPMI 1640 medium containing 10% (w / v) fetal bovine serum for 3 days and LNCaP cell line containing 10% CS-FCS (charcoal-stripped FCS, Life Technologies). After culturing for another 2 days by replacing with red-free RPMI 1640 medium, the conventional method was performed using a vector for a luciferase reporter assay (pGL3-PSA: Yonsei University Medical School) containing a PSA (prostate-specific antigen) regulatory promoter site. According to the transfection (transfection) was performed. 50 nM of synthetic androgen R1881 (17β-hydroxy-17α-methyl-19-norandrost-4,9,11-trien-3-one, Perkin Elmer) was treated and treated with 100 μg / mL RMB-1 after 48 hours. After 18 hours, reporter analysis was performed using lysates of cells. At the same time, flutamide (final concentration: 20 μM), already widely known as a prostate cancer drug, was treated alone or in parallel with 100 μg / mL of RMB-1.

As shown in FIG. 6, RMB-1 inhibited transcription to a level similar to that of the flutamide treated group at a concentration of 100 µg / mL, and when RMB-1 and flutamide were simultaneously treated, it was suppressed to the control level. Synergy could be confirmed.

Example 6 Inhibition of mRNA Expression of Androgen Receptor Regulator Genes of Glycosylated Stem Butanol Fractions in Prostate Cancer Cell Lines

LNCaP cell lines were cultured in RPMI 1640 medium containing 10% (w / v) fetal bovine serum for 3 days, and LNCaP cell lines were exchanged with phenol red-free RPMI 1640 medium containing 10% CS-FCS (charcoal-stripped FCS, Life Technologies). After culturing again for 3 days, the synthetic androgen R1881 at a concentration of 50 nM was treated. After 6 hours, the total concentration of RNA was extracted and treated with the corresponding stem butanol fraction at a final concentration of 100 µg / mL, and 16 hours later, RT-PCR was performed. The target gene primers involved are as follows:

① NKX 3-1:

Forward primer (SEQ ID NO: 1): 5'-AGCCGCTCACGTCCTTCCTCATCC-3 '

Reverse primer (SEQ ID NO: 2): 5'-GGGGCCCGGTGCTCAGCTCGTCGTTCT-3 '

② TSC22:

Forward primer (SEQ ID NO: 3): 5'-ATTTTTCTCTATTAGTTCTTTGATTTG-3 '

Reverse primer (SEQ ID NO: 4): 5'-GACTTGATAATAGCTCCTCTGGT-3 '

③ PSA:

Forward primer (SEQ ID NO: 5): 5'-GCCCACCCAGGAGCCAGCACT-3 '

Reverse primer (SEQ ID NO: 6): 5'-GGCCCCCAGAATCACCCGAGCAG-3 '

④ GAPDH:

Forward primer (SEQ ID NO: 7): 5'-CGCGGGGCTCTCCAGAACATCATCC-3 '

Reverse primer (SEQ ID NO: 8): 5'-CTCCGACGCCTGCTTCACCACCTTCTT-3 '

94 DEG C, 2 minutes with the above primer; 27 cycles of 94 ° C, 30 seconds, 55 ° C, 30 seconds, 72 ° C, 30 seconds; The extension was subjected to RT-PCR at 94 ° C. for 5 minutes, resulting in no R1881 (synthetic androgen) treatment as mRNA expression of androgen receptor target genes increased by treatment with R1881 (synthetic androgen) treated with RMB-1. Decreased to the same level as the group (FIG. 7).

Example 7 Investigation of Androgen Receptor and Histone Acetylation Inhibitory Activity of Glycolytic Stem Butanol Fractions in Prostate Cancer Cell Lines

To examine the androgen receptor acetylation inhibitory activity of RMB-1, the prostate cancer cell line (LNCaP) was cultured in RPMI 1640 medium containing 10% (w / v) fetal bovine serum in a 100 mm cell culture dish. After reaching fluence, the cells were replaced with phenol red-free RPMI 1640 medium containing 10% CS-FCS (charcoal-stripped FCS, Life Technologies) and incubated for 2 more days to synthesize androgen R1881 (50nM) and RMB-1 ( Cells were harvested after 18 hours with 100 μg / mL). Immunoprecipitation was performed on acetyl lysine protein antibodies only on the supernatant obtained by crushing the harvested cells into Lysis buffer and centrifuging at 13,000 rpm for 10 minutes. After 18 hours of IP at 4 ° C., beads (A / G PLUS agarose bead, Santa Cruz) were harvested and immunoblotting was performed on the androgen receptor antibody.

As shown in FIG. 8A, the acetylation of the androgen receptor, which was increased with the treatment of R1881 (synthetic androgen), decreased to the same level as the R1881 (synthetic androgen) untreated group as RMB-1 was treated.

Next, in order to examine the regulation of histone acetylation inhibition of RMB-1, prostate cancer cell line (LNCaP) was collected and treated in the above conditions in a 150 mm cell culture dish. The harvested cells were fixed in PBS with 1% formaldehyde, treated with 100 mM Tris (pH9.4) containing 10 mM dithiothreitol for 15 minutes at 30 ° C. and then treated with Sol A buffer (10 mM Hepes pH7.9). , 0.5% NP-40, 1.5 mM MgCl ₂ , 10 mM KCl, 0.5 mM DTT). After centrifugation at 800 × g for about 5 minutes, the pellet was heated with Sol B buffer (20mM Hepes pH7.9, 25% Glycerol, 0.5% NP-40, 0.42M NaCl, 1.5mM MgCl ₂ , 0.2mM EDTA) Nuclear protein was extracted by pipetting. After centrifugation at 13,000 × g for 30 minutes, the pellets were cut and soaked in IP buffer (1% Triton X-100, 2mM EDTA, 20mM Tris-HCl pH 8.0, 150mM NaCl, protease inhibitor) about 0.5 to 1kb in length. . Chromatin immunoprecipitation (ChIP) assay was performed according to methods of Shang, Y. et al. ( Cell 103, 843-852, 2000) PSA and B2M (β-2-microglobulin) primers (PSA: Forward primer (SEQ ID NO: 9): 5'-CATGTTCACATTAGTACACCTTGCC-3 ', Reverse primer (SEQ ID NO: 10): 5'-TCTCAGATCCAGGCTTACTGTC-3'B2M; Forward primer (SEQ ID NO: 11): 5'-AGACTTCCCAAATTTTGCCATCCTA-3 ', Reverse primer (SEQ ID NO: 12): 5 As a result of the antibody (IgG, αAR, αAcH3, αAcH4) of FIG. 8B with '-AAAGGCCTGAAATGTAAGTGTTGAGT-3', it was confirmed that acetylation of histones 3 and 4 was rapidly suppressed when RMB-1 was treated (FIG. 8B). .

Example 8 Investigation of Inhibitory Activity on Cell Growth and Proliferation of Glycogenated Stem Butanol Fraction in Various Cancer Cell Lines

To examine the activity of RMB-1 on cell growth and proliferation in various cancer cell lines, breast cancer cell line (MCF-7; ATCC, USA) and androgen receptor containing prostate cancer cell lines (LNCaP; ATCC, USA) were respectively 1.0 After seeding in 96 well-plates at × 10 ³ and 1.5 × 10 ⁴ cells / well, incubated at 37 ° C for 18 hours, RMB-1 was treated at a concentration of 200 μg / mL for 48 hours, followed by MTT (3- (4, 5-dimethylthiazol-2-yl) -2, 5-diphenyltetrazolium bromide: 1mg / mL) was added and reacted at 37 ° C for 2 hours, and the formed formazan was absorbed at 570nm and reference 630nm. It was confirmed by measurement.

As shown in FIG. 9, RMB-1 showed a cell viability of 24% at a concentration of 200 μg / mL, and thus an excellent anticancer effect was confirmed as compared to breast cancer (95%) cell line.

Preparation Example 1 Preparation of Tablet

The following formulation tablets (based on 200 parts by weight of total composition) were prepared by a conventional tablet press.

20 parts by weight of powder of the corresponding stem extract

Dextrin 72 parts by weight

80 parts by weight per minute

8 parts by weight of glycerin fatty acid ester

Mixing and tableting of raw materials were easy, and refined refinement | purification was obtained.

Preparation Example 2 Preparation of Capsule

By the usual method, the capsule which has the following compositions was manufactured. In addition, No. 1 hard gelatin capsule was used for the capsule.

<Composition in 1 capsule (200mg per tablet)>

Powder 5mg of Glycerium Stem Extract

Cornstarch 60.0mg

Lactose 100.0 mg

Calcium Lactate 10.0mg

Hydroxypropyl cellulose (HPC-L) 10.0 mg

Preparation Example 3 Preparation of Granules

By conventional methods, instant tea granules of the following formulations were prepared by fluid bed granulator (based on 970 parts by weight total composition).

20 parts by weight of powder of the corresponding stem extract

40 parts by weight of oligosaccharide

50 parts by weight of citric acid

50 parts by weight of sugar

Dextrin 810 parts by weight

Mixing of the raw materials and granulation by a fluidized bed granulator were easy and fine tea granules having good taste were obtained.

Production Example 5 Beverage Preparation

After dissolving 500 mg of glycolytic stem extract according to the present invention in an appropriate amount of water, vitamin C as an auxiliary component, citric acid, sodium citrate, oligosaccharide as an auxiliary agent is added, and an appropriate amount of sodium benzoate is added as a preservative, followed by adding water Was made to 100 mL to prepare a beverage. At this time, taurine, myo-inositol, folic acid, and pantothenic acid may be added alone or together.

1 is a graph showing the inhibitory effect on the HAT protein of glycolysis stem methanol extract.

Figure 2 is a graph comparing the HAT inhibitory activity of methanol extracts of glycolysis fruit, root and stem areas.

Figure 3 is a schematic of the organic solvent fraction of glycolysis stem methanol extract.

Figure 4 is a graph showing the inhibitory effect of the organic solvent fractions of glycolysis stem methanol extract on HAT protein.

5 is a graph showing the enzyme specific inhibitory effect on p300 and CBP of the butanol fraction of glycolysis stem methanol extract.

6 is a graph showing the transcriptional activity inhibiting ability of the butanol fraction of glycolytic stem methanol extract in prostate cancer cell line.

7 is a photograph showing the inhibitory effect on the mRNA expression of the androgen receptor target gene of the butanol fraction of glycolytic stem methanol extract in prostate cancer cell line.

Figure 8 is a photograph showing the inhibitory effect on the androgen receptor and histone protein acetylation of butanol fraction of glycolytic stem methanol extract in prostate cancer cell line.

9 is a graph showing the effect on the cell viability of the butanol fraction of glycolytic stem methanol extract in various cancer cell lines.

<110> Industry-Academic Cooperation Foundation, Yonsei University <120> A composition for preventing or treating cancer comprising stem          extract of Rosa rugosa <160> 12 <170> KopatentIn 1.71 <210> 1 <211> 24 <212> DNA <213> Artificial Sequence <220> 223 NKX 3-1: Forward primer <400> 1 agccgctcac gtccttcctc atcc 24 <210> 2 <211> 27 <212> DNA <213> Artificial Sequence <220> 223 NKX 3-1: Reverse primer <400> 2 ggggcccggt gctcagctcg tcgttct 27 <210> 3 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> TSC22: Forward primer <400> 3 atttttctct attagttctt tgatttg 27 <210> 4 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> TSC22: Reverse primer <400> 4 gacttgataa tagctcctct ggt 23 <210> 5 <211> 21 <212> DNA <213> Artificial Sequence <220> P223: forward primer <400> 5 gcccacccag gagccagcac t 21 <210> 6 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> PSA: Reverse primer <400> 6 ggcccccaga atcacccgag cag 23 <210> 7 <211> 25 <212> DNA <213> Artificial Sequence <220> GAPDH: Forward primer <400> 7 cgcggggctc tccagaacat catcc 25 <210> 8 <211> 27 <212> DNA <213> Artificial Sequence <220> GAPDH: Reverse primer <400> 8 ctccgacgcc tgcttcacca ccttctt 27 <210> 9 <211> 25 <212> DNA <213> Artificial Sequence <220> P223: forward primer <400> 9 catgttcaca ttagtacacc ttgcc 25 <210> 10 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> PSA: Reverse primer <400> 10 tctcagatcc aggcttactg tc 22 <210> 11 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> B2M: Forward primer <400> 11 agacttccca aattttgcca tccta 25 <210> 12 <211> 26 <212> DNA <213> Artificial Sequence <220> <223> B2M: Reverse primer <400> 12 aaaggcctga aatgtaagtg ttgagt 26  

Claims (17)

Pervasive ( Rosa rugosa ) Cancer preventive or therapeutic composition comprising the stem extract as an active ingredient. The method of claim 1, The glycolytic stem extract is water, a lower alcohol having 1 to 6 carbon atoms or a mixed solvent thereof, or a butanol fraction of the lower alcohol extract, wherein the composition for preventing or treating cancer. The method of claim 1, Effective amount of glycolysis ( Rosa rugosa ) A composition for preventing or treating cancer, characterized in that the pharmaceutical composition for the prevention or treatment of hormone receptor mediated cancer comprising a stem extract. The method of claim 3, Hormone receptor-mediated cancer is a composition for preventing or treating cancer, characterized in that the prostate cancer. The method of claim 3, A cancer prophylactic or therapeutic composition further comprising a pharmaceutically acceptable carrier. The method of claim 3, Cancer preventive or therapeutic composition further comprising flutamide (Flutamide). Pervasive ( Rosa rugosa ) Cancer preventive or improved dietary supplement comprising stem extract as an active ingredient. The method of claim 7, wherein The glycolytic stem extract is water, a lower alcohol having 1 to 6 carbon atoms or a mixed solvent thereof, or a butanol fraction of the lower alcohol extract, a cancer supplement or preventive health supplement. The method of claim 7, wherein Effective amount of glycolysis ( Rosa rugosa ) a health supplement for cancer prevention or improvement, characterized in that the food or food additive containing a stem extract. The method of claim 7, wherein Effective amount of glycolysis ( Rosa rugosa ) a health supplement for cancer prevention or improvement, characterized in that the beverage or beverage additive containing a stem extract. The method of claim 7, wherein The cancer is a cancer preventive or improved dietary supplement, characterized in that the hormone receptor mediated cancer. The method of claim 11, Hormone receptor mediated cancer is a cancer supplement for cancer prevention or improvement, characterized in that the prostate cancer. Pervasive ( Rosa rugosa ) histone acetyl transferase (HAT) inhibitor comprising a stem extract as an active ingredient. The method of claim 13, A glycolytic stem extract is a histone acetyltransferase (HAT) inhibitor, characterized in that extracted with water, a lower alcohol having 1 to 6 carbon atoms or a mixed solvent thereof, or a butanol fraction of the lower alcohol extract. The method of claim 13, Effective amount of glycolysis ( Rosa rugosa ) A histone acetyl transferase (HAT) inhibitor characterized by being an anti-inflammatory agent comprising a stem extract. The method of claim 13, Effective amount of glycolysis ( Rosa rugosa ) Histone acetyl transferase (HAT) inhibitor, characterized in that the treatment for acquired immune deficiency comprising a stem extract. The method of claim 13, A histone acetyl transferase (HAT) inhibitor further comprising flutamide.

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