TWI762787B - A kind of pale bamboo leaf extract and its monomer and anticancer use - Google Patents

Abstract

The present invention provides a pale bamboo leaf extract and a monomer thereof with a therapeutic effect on cancer, as well as the use of the pale bamboo leaf extract and the monomer for preparing a composition for treating cancer; wherein the active components of the pale bamboo leaf extract and the Monomers include orientin, vitexin, isoorientin, isovitexin, swertiajaponin, schaftoside or isovitexin Isoschaftoside.

Description

A kind of pale bamboo leaf extract and its monomer and anticancer use

The present invention relates to the leaf extracts of the pale bamboo and their monomers and uses, and particularly relates to the use of the extracts of the leaves of the pale bamboo and the monomers for the preparation of compositions for preventing or treating cancer.

Cancer is a tumor in which cells divide and proliferate abnormally, causing damage to adjacent tissues and organs, and can even metastasize to other parts of the body, so it is also called malignant tumor. Cancers are usually named after the primary site, and cancers that metastasize from the primary site to other sites are called metastatic cancers at the primary site.

Currently, more than one hundred cancers are known in humans, and cancer is one of the leading causes of death in developed countries. Cancer has become an important issue that countries all over the world, especially developed and developing countries, must face. Therefore, effective active ingredients that can be used for cancer treatment and are helpful for the development of new drugs have also become the targets actively sought by various academic circles and industries.

Lophatherum gracile ( Lophatherum gracile ), belonging to Poaceae, subfamily Aloe, and genus Lophatherum, is widely distributed in various regions of Asia. It is believed to have antipyretic, diuretic, and antibacterial properties. However, it is unknown whether it has anti-cancer properties.

The purpose of the present invention is to provide the use of a bamboo leaf extract for preparing a composition for treating cancer, wherein the bamboo leaf extract has the effect of inhibiting the growth of cancer cells.

In order to achieve the aforementioned object of the invention, the active ingredients of the pale bamboo leaf extract include orientin, vitexin and swertiajaponin.

In order to achieve the above-mentioned purpose of the invention, the active ingredients of the pale bamboo leaf extract comprise orientin, vitexin, isoorientin, isovitexin, Radix Radix et Rhizoma Swertiajaponin, schaftoside and isoschaftoside.

Another object of the present invention is to provide the use of an active monomer of eucalyptus leaves for preparing a composition for treating cancer, wherein the active monomer system of eucalyptus leaves is selected from the group consisting of orientin and vitexin. , the group consisting of isoorientin, isovitexin, swertiajaponin, schaftoside and isoschaftoside.

In order to achieve the object of the invention, wherein the active single system of bamboo leaf is selected from the group consisting of orientin, vitexin and swertiajaponin.

Another object of the present invention is to provide a method for treating or preventing cancer A composition for treating diseases, which comprises an effective amount of Bamboo leaf extract; wherein the Bamboo leaf extract has the effect of inhibiting the growth of cancer cells.

In order to achieve the aforementioned object of the invention, the active ingredients of the pale bamboo leaf extract include orientin, vitexin and swertiajaponin.

In order to achieve the above-mentioned purpose of the invention, the active ingredients of the pale bamboo leaf extract comprise orientin, vitexin, isoorientin, isovitexin, Radix Radix et Rhizoma Swertiajaponin, schaftoside and isoschaftoside.

In order to achieve the aforementioned object of the invention, the cancer is lung cancer or liver cancer.

In order to achieve the aforementioned object of the invention, the lung cancer is non-small cell lung cancer.

In order to achieve the purpose of the foregoing invention, the dosage form of the composition comprises solution, emulsion, suspension, powder, lozenge, oil, ointment, lozenge or capsule.

In order to achieve the purpose of the foregoing invention, the composition may further comprise at least one pharmaceutically acceptable carrier or salt.

In order to achieve the purpose of the foregoing invention, wherein the carrier comprises excipients, diluents, thickeners, fillers, binders, disintegrants, lubricants, oily or non-oily bases, surfactants, suspending agents, gums Coagulants, adjuvants, preservatives, antioxidants, stabilizers, colors or flavors.

The object of the present invention is also to provide a composition for the treatment or prevention of cancer, which comprises an effective amount of the active monomers of Bamboo leaves; wherein the active monomers of Bamboo leaves are selected from the group consisting of orientin, Vitex The group consisting of vitexin, isoorientin, isovitexin, swertiajaponin, schaftoside and isoschaftoside Group.

In order to achieve the object of the invention, wherein the active single system of bamboo leaf is selected from the group consisting of orientin, vitexin and swertiajaponin.

In order to achieve the aforementioned object of the invention, the cancer is lung cancer or liver cancer.

In order to achieve the aforementioned object of the invention, the lung cancer is non-small cell lung cancer.

In order to achieve the purpose of the foregoing invention, the dosage form of the composition comprises solution, emulsion, suspension, powder, lozenge, oil, ointment, lozenge or capsule.

In order to achieve the purpose of the foregoing invention, the composition may further comprise at least one pharmaceutically acceptable carrier or salt.

In order to achieve the purpose of the foregoing invention, wherein the carrier comprises excipients, diluents, thickeners, fillers, binders, disintegrants, lubricants, oily or non-oily bases, surfactants, suspending agents, gums Coagulants, adjuvants, preservatives, antioxidants, stabilizers, colors or flavors.

The present invention provides a combination that can be used to prepare a combination for the treatment of cancer novel active ingredients. The bamboo leaf extract and its monomers provided by the present invention have the activity of inhibiting the growth of cancer cells, and can inhibit the growth of cancer cells of different cancers, and are very suitable as effective components of a composition for treating cancer.

Figure 1 shows the inhibitory effect of the bamboo leaf extract on the growth of cancer cells.

Fig. 2 shows the inhibitory effect of Huh-7 cell growth of the active fractions of the leaf bamboo.

Figure 3 shows the inhibitory effect of H520 cell growth of the active fractions of the leaf bamboo.

Fig. 4 shows the results of the analysis of the monomer components of the active fractions of Pseudomonas sinensis.

All technical and scientific terms used in this specification, unless otherwise defined, have the meaning commonly understood in the field of professionals involved, wherein the singular terms "a", "an", "the", "said" ", unless otherwise stated, can refer to more than one object, and the terms "include" and "include" are open-ended conjunctions; in addition, unless otherwise stated, the materials used in this case are commercially available and readily available.

As used in this specification, the term "extract" refers to a product prepared by extraction. The extract may be presented as a solution in a solvent, or the extract may be presented as a concentrate or essence free or substantially free of solvent. The extract can also be formulated into pharmaceutical compositions or foods. The term extract may be a single extraction obtained from a specific extraction step or a series of extraction steps The extract, or the extract can also be a combination of extracts obtained from separate extraction steps. Accordingly, these combined extracts are also encompassed by the term "extract".

The term "treating" refers to a method of delaying, ameliorating, reducing or reversing the disorder or any symptoms associated with the disorder currently afflicting a patient and methods of preventing the disorder or any emerging symptoms thereof.

The term "pharmaceutically acceptable" means that a substance or composition must be compatible with the other ingredients of the formulation and not injurious to the patient.

The term "pharmaceutically acceptable excipient", as used herein, means any physiologically inert, pharmacologically inert substance known to those skilled in the art to be compatible with the physical and chemical properties of the extract active substance. Pharmaceutically acceptable excipients include, but are not limited to, polymers, resins, plasticizers, fillers, lubricants, diluents, binders, disintegrants, solvents, co-solvents, surfactants, preservatives , sweeteners, flavoring agents, pharmaceutical grade dyes or pigments, and viscosity agents.

The pale bamboo leaf used in the embodiment of the present invention is Lophatherum gracile , which can be purchased in the market. The embodiments of the present invention are exemplified by Lophatherum gracile , but the present invention is not limited by Lophatherum gracile . The species covered by the genus Lophatherum have the same active monomer components as the Lophatherum leaves used in the embodiments of the present invention. All species are intended to be included within the scope of the present invention.

The examples of the present invention are illustrated by the human lung cancer cell line (H520) and the human liver cancer cell line (Huh-7), but the present invention is not limited to human lung cancer Cell line (H520) and human hepatoma cell line (Huh-7). The H520 cell line was purchased from Food Industry Development Research Institute (BCRC); the Huh-7 cell line was purchased from JCRB (Japanese Collection of Research Bioresources Cell Bank).

The H520 cell line in the cell experiment of the embodiment of the present invention uses the RPMI-1640 medium containing 10% fetal bovine serum; the Huh-7 cell line uses the DMEM high glucose medium containing 10% fetal bovine serum, and is cultured in the following manner: Cells were seeded in T75 cell culture flasks and cultured in a 37°C, 5% _CO2 incubator. When the cells grow to 90% of the T75 cell culture flask, wash the cells once with PBS, add Trypsin-EDTA to detach the cells, and replace them with a medium containing 5% fetal bovine serum, and seed 6×10 ³ cells into a 96-well culture. in a dish (volume: 180 μl/well), then cultured in a 37° C., 5% CO ₂ incubator for 4 hours, and set aside.

The present invention is based on the following examples, but the present invention is not limited by the following examples.

Example 1. Preparation of Bamboo Leaf Extract

A pale bamboo leaf extract (LGCH) was obtained by extracting a pale bamboo leaf with an alcohol solvent. Then, the extracts of the pale bamboo leaves were separated into the active fractions (LGCH1-1, LGCH1-2, LGCH2, LGCH3, LGCH4 and LGCH5) of the pale bamboo leaves using a macroporous resin column and ethanol-water gradient elution, and were made into the active fractions of the pale bamboo leaves. The lyophilized product was detected and analyzed by UPLC for its monomer composition. Wherein, the alcoholic solvent includes but is not limited to methanol, ethanol or propanol.

cell experiment

Example 2. Test of anticancer effect of the extract of pale bamboo leaves

Cell treatment method: take the pale bamboo leaf extract (LGCH) described in Example 1, use 100% DMSO to prepare it into a 50 mg/mL pale bamboo leaf extract stock solution, and then use a medium containing 5% fetal bovine serum. Diluted to different concentrations (2000, 1500, 1000, 500, 250, 125, 62.5, 31.25 μg/mL) of the dilute bamboo leaf extract. Take 20 μl of the aforementioned diluents of different concentrations of Bamboo leaf extract, respectively, add them to the cells (final volume: 200 μl/well), and culture at 37 °C and 5% CO ₂ for 48 hours, and then analyze the cell viability.

Cell viability assay (MTS assay): After culturing the cells of the different treatments for 48 hours, the medium containing the diluent of Bamboo leaf extract was removed, and then 100 μl of MTS reagent (using 5% fetal bovine serum) was added to each well. After 1 hour of reaction in a 37°C, 5% CO ₂ incubator, the absorbance at 490 nm was analyzed using an ELISA reader.

The results of the analysis are shown in Table 1 and Figure 1, and the LGCH can inhibit the growth of cancer cells, and its half maximal inhibitory concentration (IC ₅₀ ) on Huh-7 cells is 160.69 μg/mL , while the IC50 on H520 cells was 157.04μg/mL.

Embodiment 3, the anticancer effect test of the active distinguishing substance of the bamboo leaf

Cell treatment method: take the active distinguishing substance of bamboo leaf described in Example 1, use secondary water (for LGCH1-1, LGCH1-2 and LGCH2) or 100% DMSO (for LGCH3, LGCH4 and LGCH5) The active fraction was formulated into a 50 mg/mL stock solution of the active fraction of bamboo leaf, and then diluted into different concentrations (2000, 1500, 1000, 500, 250, 125, 62.5, 31.25 μg with a medium containing 5% fetal bovine serum). /mL). Take 20 μl of the above-mentioned different concentrations of the active fractions of the bamboo leaf, respectively, add them to the cells (final volume 200 μl/well), and culture at 37 ° C and 5% CO ₂ for 48 hours, and then analyze the cell viability.

Cell viability assay (MTS assay): After 48 hours of culturing the cells with different treatments, remove the medium containing the diluent of the active fractions of Pseudomonas serrata, and then add 100 μl of MTS reagent (using 5% fetal bovine) to each well. culture medium dilution of serum), and after reacting in a 37°C, 5% _CO2 incubator for 1 hour, the absorbance at 490nm was analyzed using a plate reader.

The results of the analysis are shown in Table 2, Figure 2 and Figure 3. The active fractions LGCH3, LGCH4 and LGCH5 have excellent inhibitory effects on the growth of Huh-7 cells and H520 cells, and their IC _50s are all within 30~41μg /mL.

Table 2. The IC ₅₀ of the active fractions of the leaf bamboo leaf acting on Huh-7 cells and H520 cells

Example 3. Analysis of the monomer components of Bamboo Leaf Extract

UPLC detection and analysis: Ultra-high performance liquid chromatography (UPLC, model: Waters UPLC H Class) was used to detect and analyze the active monomer components of the leaf extract. Take the above-mentioned lyophilized products of the active fractions (LGCH3, LGCH4 and LGCH5) of the above-mentioned pale bamboo leaves with anticancer effect, prepare them into 1 mg/ml with methanol/water (80/20, V/V), and take 10 μl for injection (Column: Acquity UPLC BEHC18 1.7um 2.1x100mm), analyzed at 1ml/min flow rate with the following conditions: Column Temperature: 35℃; 2D channels: 331nm and 350nm; Mobile phase A : 0.1% Acetic acid; Mobile phase B: Acetonitrile.

The analysis results are shown in Figure 4 and Table 3. The active fractions LGCH3, LGCH4 and LGCH5 with anticancer activity isolated from the extracts of the leaves of the leaves of Eucalyptus sinensis contain orientin, Vitex Vitexin, isoorientin, isovitexin, swertiajaponin, schaftoside, isoschaftoside.

Orientin

swertijaponin

Vitexin

isoorientin

isovitexin

schaftoside

The present invention solves the problem that no one has used the bamboo leaf extract and its monomers as a composition for the preparation of cancer treatment. The efficacy of growth, and no significant difference in drug efficacy among cancer species, is very suitable for preparing a composition for treating cancer.

From the contents disclosed in the preferred embodiments of the present specification, those skilled in the art of the present invention can clearly understand that the foregoing embodiments are only examples; It does not differ from the technical features of the present invention. According to the embodiments of the description, the present invention can be modified in various ways without hindering the implementation. The claims provided in this specification define the scope of the invention, which encompasses the foregoing methods and structures and inventions equivalent thereto.

The above-mentioned functions are indeed in full compliance with the statutory patent requirements of novelty and progress, and you can file an application in accordance with the law.

Claims (10)

The utility model relates to the use of a pale bamboo leaf extract for preparing a composition for treating lung cancer or liver cancer, wherein the pale bamboo leaf extract has the effect of inhibiting the growth of cancer cells, and the active ingredient of the pale bamboo leaf extract is a daily medicine flavin. The use as described in item 1 of the scope of the application, wherein the flavonoids account for 47.36 wt% of the bamboo leaf extract. According to the use as described in item 1 or 2 of the patented scope, the components of the pale bamboo leaf extract may further comprise orientin, vitexin, isoorientin and isovitexin. The use as described in claim 3, wherein the orientin accounts for 1.90 wt % of the pale bamboo leaf extract, the vitexin accounts for 1.57 wt % of the pale bamboo leaf extract, and the isoorientin accounts for 1.57 wt % of the pale bamboo leaf extract 0.75 wt % of the extract, the isovitexin accounted for 4.01 wt % of the pale bamboo leaf extract. The use as described in item 3 of the claimed scope, wherein the composition may further comprise schaffoside or isoschaffatin. A composition for treating lung cancer or liver cancer, which comprises an effective amount of Bamboo leaf extract; wherein the Bamboo leaf extract has the effect of inhibiting the growth of cancer cells, and the active ingredient of the Bamboo Leaf extract is daily medicine flavin, and the The flavonoids accounted for 47.36% by weight of the pale bamboo leaf extract; wherein the components of the pale bamboo leaf extract further comprise orientin, vitexin, isoorientin and isovitexin; the orientin accounted for the pale bamboo leaf 1.90 wt % of the extract, the vitexin accounted for 1.57 wt % of the pale bamboo leaf extract, the isoorientin accounted for 0.75 wt % of the pale bamboo leaf extract, and the isovitexin accounted for 4.01 wt % of the pale bamboo leaf extract. The composition of claim 6, wherein the lung cancer is non-small cell lung cancer. The composition according to item 6 or 7 of the claimed scope, wherein the dosage form of the composition comprises solution, emulsion, suspension, powder, lozenge, oil, ointment, lozenge or capsule. The composition as described in claim 6 or 7, wherein the composition may further comprise at least one pharmaceutically acceptable carrier or salt. The composition of claim 9, wherein the carrier comprises excipients, diluents, thickeners, fillers, binders, disintegrants, lubricants, oily or non-oily bases, surface Active agent, suspending agent, gelling agent, adjuvant, preservative, antioxidant, stabilizer, color or flavor.

Images