Banana extract and use of banana extract

Abstract

To provide a hair health care composition.SOLUTION: The hair health care composition contains an effective amount of banana flower extract and a carrier, wherein the banana flower extract is a compound I represented by formula (I), a compound II represented by formula (II) or a combination thereof.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present invention relates to a banana extract and uses of the banana extract, and in particular to a banana extract for hair health care and uses of the banana extract. [Background technology]

[0002] Bananas are a tropical fruit that are popular not only for eating fresh but also for various desserts. Currently, Taiwan has approximately 10,000 hectares of banana planting land, and the abundant production is sold domestically and also exported overseas.

[0003] In addition to the male and neutral flowers at the front, banana flowers also have female flowers, but only the ovary at the bottom of the female flower can develop into a fruit after it has grown. To avoid competing for the nutrients that will become the banana fruit, the male flowers are often thinned out. The thinned-out male flowers are also called banana flowers. Summary of the Invention [Problem to be solved by the invention]

[0004] It is believed that the reasons why men are more likely to go bald include genetic factors, constitution, and hormones.

[0005] Testosterone, a male hormone present in the human body, is converted into dihydrotestosterone (DHT) by the action of reductase, which causes hair to gradually thin and fall out, and in the long term, even causes hair follicles to shrink.

[0006] Common baldness medications such as fanasteride are known to be effective in preventing the worsening of male pattern baldness, but they also have side effects such as decreased libido, erectile dysfunction, and difficulty in achieving fertility (male infertility).

[0007] In view of these circumstances, the present invention provides a treatment for baldness without side effects, and also proposes a hair health care composition and banana extract, as well as a use for preparing a hair health care composition, in order to find other uses for banana flowers that would otherwise be discarded. [Means for solving the problem]

[0008] 1. A hair care composition comprising an effective amount of banana flower extract and a carrier, wherein the banana flower extract is a compound I represented by the following formula (I), a compound II represented by the following formula (II), or a combination thereof:

[0009] [ka]

[0010] A hair care composition comprising:

[0011] 2. Also, a compound I represented by the following formula (I), a compound II represented by the following formula (II), or a combination thereof

[0012] [ka]

[0013] A banana flower extract characterized by:

[0014] 3. It is also used in the preparation of health care compositions, and the banana flower extract contains: Obtained by extracting the stamens of banana flowers through an aqueous extraction step. Use of banana flower extract.

[0015] 4. The banana flower extract also has the effect of increasing the diameter of hair roots. 3. Use of the banana flower extract according to the above item 3.

[0016] 5. The banana flower extract also has the effect of increasing the stability of hair follicles. 3. Use of the banana flower extract according to the above item 3.

[0017] 6. The banana extract also has the effect of reducing hair loss. 3. Use of the banana flower extract according to the above item 3.

[0018] 7. The banana extract is also extracted through an aqueous extraction step, a re-extraction step, and a washing step from the stamens of the banana flower. 3. Use of the banana flower extract according to the above item 3.

[0019] 8. The banana extract is a compound I represented by the following formula (I), a compound II represented by the following formula (II), or a combination thereof.

[0020] [ka]

[0021] 7. Use of the banana flower extract according to the above item 7.

[0022] 9. The banana extract also has the effect of inhibiting the production of dihydrotestosterone. 7. Use of the banana flower extract according to the above item 7.

[0023] 10. The banana extract also has the effect of promoting hair growth. 7. Use of the banana flower extract according to the above item 7.

[0024] 11. The banana extract also has the effect of increasing the proliferation of hair follicle cells. 7. Use of the banana flower extract according to the above item 7. [Effects of the Invention]

[0025] Thus, according to the present invention, any embodiment of the banana flower extract can be used to prepare a hair health care composition. The banana flower extract of any embodiment of the present invention can increase the diameter of hair roots, improve the stability of hair follicles, reduce hair loss, inhibit the production of dihydrotestosterone (DHT), promote hair growth, and increase the proliferation of hair follicle cells. [Brief explanation of the drawings]

[0026] [Figure 1] 1 is a graph showing the results of suppressing dihydrotestosterone content in a banana flower extract according to the present embodiment. [Figure 2] FIG. 1 shows the results of a banana extract according to an embodiment of the present invention promoting hair follicle cell proliferation. [Figure 3] FIG. 10 is a graph showing the results of the banana extract according to another embodiment of the present invention promoting the proliferation of hair follicle cells. [Figure 4] 1 is a graph showing the results of relative mean hair root diameter from a human study. [Figure 5] 1 is a graph showing the results of hair follicle stability from a human study. [Figure 6] 1 is a graph showing relative mean hair loss results from human studies. DETAILED DESCRIPTION OF THE INVENTION

[0027] As used herein, "banana flower" refers to the flower of the banana plant.

[0028] In an embodiment of the present invention, the banana flower extract may be extracted from banana flowers of banana (Musa sapientum L.), banana flowers of AAB silk banana (Musa spp. AAB Silk), banana flowers of pink banana (Musa spp. ABB Bluggoe), banana flowers of banana (Musa spp. AAA Robusta), and banana flowers of honey banana (Musa spp. AAB Bluggoe), or banana flowers of banana (Musa paradisiacal).

[0029] In this embodiment, the banana flowers used to extract the banana flower extract (banana flower extract) may be the stamens of the banana flower. Banana flowers come in fresh, dried, and frozen varieties.

[0030] In this embodiment, the banana flowers used to extract the banana extract may be whole banana flowers, or the banana flowers may be chopped, diced, milled, crushed, or otherwise physically processed to change the size or physical shape of the raw material from its original state.

[0031] In this embodiment, the banana extract extraction step (hereinafter also referred to as "process") includes an aqueous extraction step, and further includes a re-extraction step and a washing step.

[0032] Aqueous extraction step: The stamens of banana flowers are extracted with water to obtain an aqueous extract of banana flowers.

[0033] Re-extraction step: The aqueous extract of banana flowers is subjected to liquid phase partition extraction using water and n-butanol as solvents to obtain an n-butanol separated layer.

[0034] Elution step: The n-butanol separated layer is eluted with water and a mixed methanol solution of water and methanol, respectively.

[0035] In this embodiment, the aqueous extraction step refers to heating water to a certain temperature range, then adding banana flowers to the water, mixing, and then maintaining the temperature for a certain period of time to prepare the aqueous extract of banana flowers.

[0036] In this embodiment, the certain temperature range refers to a range between 30°C and 65°C, between 40°C and 60°C, or between 45°C and 55°C.

[0037] In this embodiment, the predetermined time period is in the range of 50 to 100 minutes. For example, banana flowers are mixed with water at 50°C and kept at 50°C for 50 minutes to obtain a water extract of banana flowers.

[0038] In this embodiment, in the aqueous extraction step, the weight ratio of water to banana flower is 2-11:1-4. For example, the ratio of water to banana flower may be 4:1.

[0039] In this embodiment, the aqueous extract of banana flowers can be obtained by subjecting the primary extract obtained by extracting the stamens of banana flowers with water in the aqueous extraction step to at least one of a centrifugation step and a filtration step. The centrifugation step refers to a step of centrifuging the initial extract to obtain a supernatant.

[0040] In this embodiment, the filtration step refers to passing the supernatant (or primary extract) obtained from the centrifugation step through a screen to filter out solids in the solution and obtain a filtrate. For example, the screen may be a 350 mesh screen. That is, the aqueous extract of banana flower may be the primary extract, the supernatant obtained from the centrifugation step, or the filtrate obtained from the filtration step.

[0041] In this embodiment, the ratio of n-butanol to water in the re-extraction step may be 3:1.

[0042] In this embodiment, the flushing step may be a secondary flush, in which the first flushing step may be a bioactivity-induced separation using a third-order linear gradient of water and methanol on an n-butanol separation layer to obtain a pure water separation section and a methanol-water solution separation section.

[0043] The secondary flush can be reversed-phase high-performance column chromatography using a linear elution gradient from pure water to 50% methanol in the water separation section, and reversed-phase high-performance column chromatography with a linear elution gradient from pure water to 100% methanol in the methanol-water separation section.

[0044] In this embodiment, the banana extract is a compound I represented by the following formula (I), a compound II represented by the following formula (II), or a combination thereof:

[0045] [ka]

[0046] Of these, compound I is N-β-citroyldopamine, and compound II is 6,2',3',6'-O-tetraacetyl-3-O-trans-p-coumaroylsucrose.

[0047] In this embodiment, the banana flower extract can be an aqueous extract of banana flower, Compound I, Compound II, or a combination of Compound I and Compound II.

[0048] In this embodiment, the effective dose of banana flower extract is 1 gram per day.

[0049] Also in this embodiment, banana flower extract can be used to prepare a hair care composition.

[0050] In this embodiment, the composition may be a health food (supplement), in other words, the supplement contains an effective amount of banana flower extract.

[0051] In this embodiment, the aforementioned health care composition can be prepared into a dosage form suitable for enteral or oral administration using techniques well known to those skilled in the art.

[0052] These dosage forms include, but are not limited to, tablets, troches, pills, capsules, dispersible powders or granules, solutions, suspensions, emulsions, syrups, elixirs, thick slurries, and the like.

[0053] In this embodiment, the healthcare composition can be prepared into a dosage form suitable for parenteral or topical administration using techniques well known to those skilled in the art, including, but not limited to, injections, sterile powders, external preparations, etc.

[0054] In this embodiment, the health care composition may be administered by a parenteral route selected from the group consisting of:

[0055] Examples of modes of administration include subcutaneous injection, intraepidermal injection, intradermal injection, and intralesional injection.

[0056] In this embodiment, the healthcare composition may further comprise a pharmaceutically acceptable carrier commonly used in the food manufacturing arts.

[0057] For example, a pharmaceutically acceptable carrier may include one or more of the following agents: solvent, buffer, emulsifier, suspending agent, decomposer, disintegrating agent, dispersing agent, binding agent, excipient, stabilizing agent, chelating agent, diluent, gelling agent, preservative, wetting agent, lubricant, absorption delaying agent, liposome, and the like.

[0058] The selection and amounts of these reagents are within the expertise and common skill of one of ordinary skill in the art.

[0059] In this embodiment, the food acceptable carrier comprises a solvent selected from the group consisting of water, normal saline, phosphate buffered saline (PBS), and an aqueous solution containing alcohol.

[0060] In this embodiment, the health care composition may be an edible composition, which may be manufactured into a food product or may be a food additive.

[0061] It means that the substance is added when preparing food ingredients using conventional methods to produce food, or added during the food manufacturing process.

[0062] Here, food may be a product containing edible ingredients for human or animal consumption.

[0063] In this embodiment, the food products may be, but are not limited to, beverages, fermented foods, bakery products, health foods, and dietary supplements.

[0064] Example 1: Preparation of banana flower extract First, bananas (scientific name: Musa paradisiacal) from Taiwan are used, and the stamens are collected from the banana flowers.

[0065] Next, water is used as a solvent, heated to 50±5°C, and then banana flowers are added. The weight ratio of banana flowers to water is 1:4. The banana flowers are mixed with water and extracted at 50±5°C for 50 minutes to obtain the initial extract.

[0066] The first extract is then filtered through a 350 mesh sieve to obtain a water extract of banana blossoms.

[0067] Next, 10 liters of the above banana flower water extract is taken and subjected to liquid phase partition extraction using n-butanol and water in a ratio of 3:1 to obtain an n-butanol layer extract and an aqueous layer extract.

[0068] Here, highly polar substances in the aqueous extract of banana flowers remain in n-butanol to form the n-butanol layer extract, and water-soluble substances remain in water to form the aqueous layer extract.

[0069] The n-butanol extract is then concentrated and dried under reduced pressure to give 24.6 g of n-butanol extract (BUF), and the aqueous extract is concentrated and dried under reduced pressure to give 198.7 grams of aqueous extract (WF).

[0070] Based on this, it can be calculated that 10 liters (L) of banana flower extract will yield a total of 223.3 grams of powdered extract, of which 11% will be n-butanol extract (BUF) and 89% will be water extract (WF).

[0071] Next, using bioassay guided fractionation, 20 grams (g) of the n-butanol layer extract is taken and subjected to macroporous resin column chromatography (Diaion HP-20 column chromatography 60 x 5 cm).

[0072] The eluents used were pure water, a methanol solution (methanol / pure water = 50 / 50), and 100% methanol. By sequential extraction, three fractions were obtained: BU1 fraction, BU2 fraction, and BU3 fraction.

[0073] In the BU1 separation section, separation is performed using reversed-phase high-performance column chromatography (RP-MPLC), with elution sequentially from water to a 50% methanol solution (the elution gradient is a linear gradient from pure water for 0 minutes to a 50% methanol solution for 80 minutes), followed by thin-layer chromatography.

[0074] The detection wavelengths were 210 nm and 280 nm, the flow rate was 20 mL / min, and 30 mL was collected in each collection section. A total of 54 fractions (divisions) were obtained, and extracts that gave similar results were combined to obtain five secondary separation sections. The five secondary separation units are referred to as a BU1-1 secondary separation unit, a BU1-2 secondary separation unit, a BU1-3 secondary separation unit, a BU1-4 secondary separation unit, and a BU1-5 secondary separation unit, respectively.

[0075] The BU1-3 secondary separation portion is then separated and purified by HPLC (detection wavelength 210 nm), and purified on a reversed-phase carbon 18 column using 20% ​​methanol as the mobile phase to obtain compound I.

[0076] Compound I,1 H, 13 C, 2D-NMR (including HSQC, HMBC, COSY, NOESY) and HR-LC-ESI-MSMS (measured simulated molecular weight m / z 350.0845 [M+Na] + , the theoretical molecular formula is C 14 H 16 NO8, theoretical simulated molecular weight m / z 350.0840, actual molecular formula C 14 H 17 NO8) was identified as N-β-citroyldopamine having the chemical structure of formula (I) below.

[0077] [ka]

[0078] In the BU2 separation section, reversed-phase high performance column chromatography (RP-MPLC) is used for separation, and the column is washed sequentially from water to methanol (the elution gradient is a linear gradient from pure water for 0 min to 100% methanol for 120 min).

[0079] Next, thin-layer chromatography was used, with detection wavelengths of 210 nm and 280 nm and a flow rate of 20 mL / min, and 30 mL was collected in each collection zone.A total of 80 collection zones were prepared, and extracts with similar results were combined.

[0080] Then, 10 secondary separation units are obtained, which are BU2-1 secondary separation unit, BU2-2 secondary separation unit, BU2-3 secondary separation unit, BU2-4 secondary separation unit, BU2-5 secondary separation unit, BU2-6 secondary separation unit, BU2-7 secondary separation unit, BU2-8 secondary separation unit, BU2-9 secondary separation unit, and BU2-10 secondary separation unit, respectively.

[0081] The BU2-8 secondary separation fraction is then purified by HPLC (detection wavelength 310 nm) and then purified on a reversed-phase carbon-18 column using 45% methanol as the mobile phase to obtain Compound II.

[0082] Compound II is 1 H, 13 C, 2D-NMR and HR-LC-ESI-MSMS (actual simulated molecular weight is m / z 655.1888 [M+H] + The theoretical molecular formula is C 29 H 35 O 17 The theoretical molecular weight is m / z 655.1880, and the actual molecular formula is C 29 H 36 O 17 ) was confirmed to be 6,2',3',6'-oxy-tetraacetyl-3-oxy-trans-p-aromatic (6,2',3',6'-O-tetraacetyl-3-O-trans-p-coumaroylsucrose) and has the chemical structure of formula (II) below.

[0083] [ka]

[0084] Here, the aqueous extract of banana flowers, Compound I and Compound II are banana flower extracts.

[0085] Example 2: Test to suppress dihydrotestosterone with banana flower extract In young male baldness patients, a significant increase in dihydrotestosterone (DHT) production is observed in the forehead hair follicles. Therefore, if the increase in DHT can be suppressed, hair follicles can be protected and hair growth can be promoted.

[0086] The study measured the reduction in dihydrotestosterone content in human prostate cells after treatment with various concentrations of banana extract and compared it with the commonly used male pattern baldness treatment, fanasteride.

[0087] Material and equipment description: Cell line: Human prostate cells LNcap (purchased from BCRC; storage number Cat. 60088), hereafter referred to as LNcap cells.

[0088] Cell culture medium (human prostate LNcap cell culture medium): RPMI 1640 medium (RPMI), powder (Gibco; Cat. 31800-022), 10% fetal bovine serum (FBS) (Gibco; Cat. 10437-028), 1% antibiotic-antimycotic (AA) (Gibco; Cat. 15240-062) was used.

[0089] The reagents were 1X DPBS (Gibco, Cat. 14200-075), trypsin (1X Trypsin-EDTA, purchased from Thermo, Cat. FNN0011), finasteride (also known as Finasteri or Finasteride, purchased from Sigma, Cat. F1293), testosterone (purchased from Sigma; Cat. T1500), and cell lysis buffer (RIPA Lysis and Extraction Buffer, purchased from Thermo, #89900).

[0090] The test kit is a dihydrotestosterone ELISA reagent test kit (USCN; CEA443Ge).

[0091] The testing process is as follows:

[0092] First, LNcap cells were cultured in a 6-well cell culture plate at 1 x 10 cells per well. 5 The cells are transplanted at a density of 1000 μg / ml and cultured at 37°C in a 5% carbon dioxide incubator for 24 hours.

[0093] The cultured LNcap cells were divided into experimental group A, experimental group B, control group, and blank group. Experimental group A was supplemented with 10 μg / mL of compound I prepared in Example 1, and experimental group B was supplemented with 5 μg / mL of compound I. The control group was supplemented with 20 μg / mL of Rope, and the blank group was supplemented with cell culture medium alone.

[0094] Next, 10 mg / mL testosterone was added to each group, and the mice were placed in a carbon dioxide incubator at 37°C for 2 hours.

[0095] After removing the supernatant from each group, wash the culture plate twice with 1X DPBS and add 200 μL of trypsin. Incubate with the LNcap cells for 3 minutes in a 37°C, 5% carbon dioxide incubator. Gently tap the cell culture plate to suspend the LNcap cells.

[0096] Next, 600 μL of cell culture medium was added to each well to stop the trypsin activity. The cell solution from each well was collected and transferred to a 1.5 mL microcentrifuge tube and centrifuged at 400 × g for 5 minutes. The supernatant was removed from the microcentrifuge tube, and 200 μL of 1X DPBS was added to the tube to wash the LNcap cells. The microcentrifuge tube was centrifuged again for 5 minutes and the supernatant was removed.

[0097] Next, add 200 μL of 1X cell lysis buffer to the tube to lyse the LNcap cells. Centrifuge at 13,000 rpm for 5 minutes at 4°C, collect the supernatant, and store it in a 1.5 mL microcentrifuge tube. Finally, use the Dihydrotestosterone ELISA Reagent Detection Kit to detect the dihydrotestosterone content in the LNcap cells. (For detailed instructions, refer to the factory instructions.)

[0098] Test results: Here, a one-tailed Student's t-test is used to analyze the two comparison groups to determine whether there is a statistically significant difference and obtain a p-value. Note that in the figure, "*" indicates a p-value less than 0.05, and "**" indicates a p-value less than 0.01. The more "*" there are, the more statistically significant the difference.

[0099] See Figure 1. The dihydrotestosterone content of the blank group was 32.8 pg / mL. The dihydrotestosterone content of the control group was 17.16 pg / mL. The dihydrotestosterone content of experimental group A was 25.03 pg / mL, and the dihydrotestosterone content of experimental group B was 25.20 pg / mL.

[0100] From this, it can be seen that there is a statistically significant difference between experimental group A and experimental group B, which used lopey or banana extract, compared to the blank group. In other words, it is clear that there is an effect of suppressing dihydrotestosterone.

[0101] Furthermore, the effectiveness of banana extract B at a concentration of 5 μg / mL could produce a significant difference compared to the blank group. At a concentration of 10 μg / mL, banana extract B showed a significant effect similar to that of the commercially available drug Ropepa, with a p-value of less than 0.01. Thus, it can be seen that banana flower extract has health benefits for hair.

[0102] Example 3: Test to see whether banana flower extract promotes hair follicle cell proliferation Hair follicle cells are the basis of hair, and during the growth phase, cells in the hair follicle bulb at the root can divide once a day. Promoting hair follicle cell proliferation can promote hair growth. In this study, the proliferation of human hair follicle cells treated with Compound I was investigated.

[0103] Material and equipment description: Cell line: Human Follicle Dermal Papilla Cells (HFDPC) (purchased from PromoCell; Cat. C-12071), hereinafter also referred to simply as "HFDPC cells."

[0104] Cell culture medium: Follicle Dermal Papilla Cell Growth Medium (purchased from PromoCell; Cat. C-26501).

[0105] Reagents: 10X DPBS (Gibco, Cat. 14200-075), trypsin (1X Trypsin-EDTA, purchased from Thermo, Cat. FNN0011).

[0106] The test kit contained Composition C (Dimethyl sulfoxide DMSO), Composition D (Click-iT TM fixative), composition E (Click-iT TM saponin-based permeabilization and wash reagent), composition F (Copper protectant), and composition G (Click-iT TM Cell proliferation ELISA reagent test kit (Click-iT) containing EdU buffer additive TM Plus EdU Flow Cytometry Assay Kits-Alexa Fluor TM 488 picolyl azide, 50 tests Brand Invitrogen Model C10632).

[0107] Testing process: First, HFDPC cells were plated in a 6-well cell culture plate at 1 x 10 cells per well. 5 The cells are then cultured in a 5% carbon dioxide incubator at 37°C for a further 24 hours.

[0108] The cultured HFDPC cells were divided into an experimental group and a blank group, in which 10 μg / mL of Compound I prepared in Example 1 was added to the experimental group, and only the cell culture medium was added to the blank group.

[0109] Next, 10 μM of composition C is added to each group, and then the cells are placed in a carbon dioxide incubator at 37° C. for 2 hours.

[0110] After removing the cell culture medium from each group, wash the culture plate once with 1X DPBS. Add 200 μL of trypsin and incubate the HFDPC cells at 37°C in a 5% carbon dioxide incubator for 5 minutes. Then, gently tap the cell culture plate to suspend the HFDPC cells.

[0111] Subsequently, the supernatant is removed, and 200 μL of cell culture medium is added to each well to stop the action of trypsin.

[0112] Next, the cell solution in each well was collected, placed in a 1.5 mL microcentrifuge tube, and centrifuged at 400 × g for 5 minutes. The supernatant was removed from the microcentrifuge tube, washed with 1 × DPBS, and centrifuged (400 × g for 5 minutes). The supernatant was removed, and 100 μL of Composition D was added to the tube. The tube was then stored at room temperature for 10 minutes in the dark.

[0113] Next, the supernatant was removed, the tube was washed again with 1X DPBS, and centrifuged (400 x g for 5 minutes). After removing the supernatant, 100 μL of Composition E was added to the tube and stored in the dark at room temperature for 15 minutes. Next, 300 μL of 1% BSA / PBS washing buffer was added, and the tube was centrifuged at 400 x g for 5 minutes. (This step was repeated three times.)

[0114] After removing the supernatant, add 100 μL of Composition E to the tube and store it in the dark at room temperature for 15 minutes. TM 100 μl of the reagent (configured based on the instruction manual of the cell proliferation ELISA reagent detection kit) is added to each centrifuge tube and allowed to react at room temperature in the dark for 30 minutes.

[0115] After adding 300 μl of 1% BSA / PBS washing buffer, the mixture is centrifuged at 400×g for 5 minutes, and then the fluorescence signal is measured using a flow cytometer (excitation light: 488 nm, scattering light: 527 nm & 590 nm).

[0116] Test results: Here, a one-tailed Student's t-test was used to analyze the two comparison groups to determine whether there was a statistically significant difference, and a p-value was obtained. In the figure, an "*" indicates a p-value less than 0.05, and an "**" indicates a p-value less than 0.01. The more "*" there are, the more statistically significant the difference.

[0117] Referring to Figure 2, if the number of HFDPC cells in the blank group is taken as 100%, the equivalent number of HFDPC cells in the experimental group is 118.81%. It was found that Compound I had a statistically significant difference compared to the blank group, demonstrating that banana flower extract has the effect of promoting hair follicle cell proliferation.

[0118] Example 4: Test to see if banana flower extract promotes hair follicle cell proliferation Hair follicle cells are the basis of hair, and during the growth phase, cells in the hair follicle bulb at the root can divide once a day. Promoting hair follicle cell proliferation can promote hair growth. This study examines the proliferation of human hair follicle cells treated with Compound II.

[0119] Material and equipment description: Cell line: Human hair follicle dermal papilla cells (HFDPC) (purchased from PromoCell; Cat. C-12071), hereinafter referred to as HFDPC cells.

[0120] Cell culture medium: Follicle Dermal Papilla Cell Growth Medium (purchased from PromoCell, Cat. C-26501).

[0121] Reagents: 10X DPBS (purchased from Gibco, Cat. 14200-075), trypsin (1 X Trypsin-EDTA, purchased from Thermo, Cat. FNN0011).

[0122] Test Kit: Cell Proliferation ELISA Reagent Test Kit (Click-iT TM Plus EdU Flow Cytometry Assay Kits-Alexa Fluor TM 488 picolyl azide, 50 tests (Brand: Invitrogen Model C10632), Composition C (Dimethyl sulfoxide DMSO), Composition D (Click-iT TM fixative), composition E (Click-iT TM saponin-based permeabilization and wash reagent), Composition F (Copper protectant), Composition G (Click-iT TM EdU buffer additive).

[0123] Testing process: First, HFDPC cells were plated in a 6-well cell culture plate at 1 × 10 5 The cells are then cultured in a 5% carbon dioxide incubator at 37°C for 24 hours.

[0124] The cultured HFDPC cells were divided into an experimental group, a (positive) control group, and a blank group, where the experimental group was treated with 100 μM of Compound II prepared in Example 1, the control group was treated with 20% FBS fetal bovine blood, and the blank group was treated with cell culture medium alone.

[0125] Next, 10 μM of Composition C was added to each group, which was then placed in a carbon dioxide incubator at 37° C. for 2 hours. After removing the cell culture medium from each group, the culture plate was washed once with 1×DPBS, 200 μL of trypsin was added, and the HFDP cells were incubated in a 5% carbon dioxide incubator at 37° C. for 5 minutes, and the cell culture plate was gently tapped to suspend the HFDPC cells.

[0126] Subsequently, the supernatant is removed, and 200 μL of cell culture medium is added to each well to stop the action of trypsin.

[0127] Next, the cell solution from each well is collected and transferred to a 1.5 mL microcentrifuge tube, centrifuged at 400 x g for 5 minutes, the supernatant in the microcentrifuge tube is removed, washed with 1X DPBS, centrifuged (at 400 x g for 5 minutes), the supernatant is removed, 100 μL of composition D is added to the tube, and it is stored in the dark at room temperature for 10 minutes.

[0128] Next, remove the supernatant, wash again with 1X DPBS, centrifuge (400 x g, 5 minutes), remove the supernatant, add 100 μL of Composition E to the tube, store in the dark at room temperature for 15 minutes, add 300 μL of 1% BSA / PBS wash buffer, and centrifuge at 400 x g for 5 minutes (repeated this step three times).

[0129] After removing the supernatant, add 100 μL of Composition E to the tube and store it in the dark at room temperature for 15 minutes. TM 100 μl of the reagent (configured based on the Cell Proliferation ELISA Reagent Detection Kit operating manual) was added to each centrifuge tube and incubated at room temperature in the dark for 30 minutes.

[0130] After adding 300 μl of 1% BSA / PBS washing buffer, the mixture is centrifuged at 400×g for 5 minutes, and finally, the fluorescence signal (excitation light: 488 nm, scattering light: 527 nm & 590 nm) is measured using a flow cytometer.

[0131] Test results: Here, a one-tailed Student's t-test was used to analyze the two comparison groups and obtain a p-value to determine whether there was a statistically significant difference. In the figure, "*" indicates a p-value less than 0.05, and "**" indicates a p-value less than 0.01. The more "*" there are, the more statistically significant the difference.

[0132] This will be explained with reference to Figure 3. If the number of HFDPC cells in the blank group is taken as 100%, the number of HFDPC cells in the control group is 117.01%, and the number of HFDPC cells in the experimental group is 112.54%. It was found that Compound II showed a statistically significant difference compared to the blank group, demonstrating that banana flower extract has the effect of promoting hair follicle cell proliferation.

[0133] Example 5: Human trials of banana flower extract Subjects: 50 people (25 people in the control group and 25 people in the experimental group). Each subject is an adult aged 20 or over.

[0134] Test items: hair root diameter, hair follicle stability, average hair loss after shampooing. Hair root diameter is measured using a digital external micrometer, Mitutoyo C / N293-100 model.

[0135] Of these, hair follicle stability refers to applying a pulling force to hairs (approximately 60 hairs in each area) in three locations on the frontal bone, temporal bone, and occipital bone, and counting the number of hairs that fall out. In other words, the fewer hairs that fall out when pulled, the more stable the hair follicle is.

[0136] Additionally, the average amount of hair loss after shampooing refers to the amount of hair loss observed when washing hair.

[0137] Test Method: The experimental group of 25 subjects was given a powder containing 1 g of the dried banana flower water extract prepared in Example 1 every day. The remaining 25 control subjects were given a placebo containing no banana flower extract every day for 12 weeks.

[0138] The data measured before intake (i.e., week 0) are referred to as control group 01 and experimental group 01, respectively. The data measured four weeks after intake (i.e., week 4) are referred to as control group 02 and experimental group 02, respectively. Furthermore, the data measured after eight weeks of intake (i.e., week 8) are referred to as control group 03 and experimental group 03, respectively. The data measured after 12 weeks of intake (i.e., week 12) are referred to as control group 04 and experimental group 04, respectively.

[0139] Test results: The graph below shows the average values ​​for all subjects, converted into relative values ​​for each group, with the data for week 0 set at 100%.

[0140] The standard deviation is calculated using the STDEV formula in Excel software. A one-tailed Student t-test in Excel software is used to analyze whether there is a statistically significant difference and obtain the p-value.

[0141] In addition, in the figure, "*" means that the p-value is less than 0.05, "**" means that the p-value is less than 0.01, and "***" means that the p-value is less than 0.001, indicating that there is a statistically significant difference.

[0142] The data from the control group at the same number of weeks was compared, and a t-test was used to analyze whether there was a statistically significant difference, and a p-value was calculated. In the figure, "#" means a p-value of less than 0.05, "##" means a p-value of less than 0.01, and "###" means a p-value of less than 0.001, indicating a statistically significant difference.

[0143] Referring to Figure 4, after four weeks of daily intake of banana flower extract, the average hair root diameter of the 25 experimental subjects increased from 100% (experimental group 01) at week 0 to 102.5% (experimental group 02). After eight weeks of daily intake of banana flower extract, the average hair root diameter increased to 103.2% (experimental group 03). After 12 weeks of daily intake of banana flower extract, the average hair root diameter increased significantly to 107.8% (experimental group 04).

[0144] Please continue to refer to Figure 4. The mean hair root diameter in the placebo group increased from 100% (control group 01) to 101.3% (control group 02) at week 0, and continued daily supplementation with banana flower extract until the mean hair root diameter was reached at week 8. After 12 weeks of daily supplementation with banana flower extract, the mean hair root diameter increased to 102.4% (control group 03).

[0145] Thinning hair is generally one of the first symptoms of male pattern baldness. Measuring the diameter of hair roots shows that they gradually thicken, indicating that the condition of male baldness is beginning to improve. In the experimental group of this study, the diameter of hair roots further increased from week 0 to week 12, and in particular, at week 12, it was significantly thicker by 6.4% compared to the control group. In other words, even excluding the placebo effect, daily intake of banana flower extract significantly increased the diameter of hair roots, making each hair strand healthier and providing hair health care benefits.

[0146] See Figure 5. After four weeks of daily banana flower extract intake, the hair follicle stability of 25 experimental subjects decreased from 100% at week 0 (Experimental Group 01) to 86.7% (Experimental Group 02). After eight weeks of daily banana flower extract intake, hair follicle stability decreased to 35.6% (Experimental Group 03). After 12 weeks of daily banana flower extract intake, hair follicle stability significantly decreased to 17.8% (Experimental Group 04).

[0147] Please continue to refer to Figure 5. In the placebo group, hair follicle stability increased from 100% (control group 01) at week 0 to 88.6% (control group 02). After 8 weeks of daily banana flower extract supplementation, mean hair root diameter increased to 85.7% (control group 03). After 12 weeks of daily banana flower extract supplementation, hair follicle stability decreased to 85.7% (control group 04).

[0148] In the experimental group, it was found that the number of hairs that fell out when force was applied continued to decrease significantly from week 0 to week 12. In particular, compared to the control group, the number was significantly lower at 67.9% at week 12. In other words, even after subtracting the placebo effect, daily intake of banana flower extract significantly reduced the number of hairs that fell out when force was applied, and improved the stability of hair follicles. Hair health benefits can be achieved by improving the health of each individual hair.

[0149] See Figure 6. After 12 weeks of daily intake of banana flower extract, the relative mean hair loss rate when shampooing for the 25 experimental group subjects significantly decreased from 100% (experimental group 01) at week 0 to 66.9% (experimental group 04). Continue to refer to Figure 6. The relative mean hair loss rate for the placebo group increased from 100% (control group 01) at week 0 to 123.2% (control group 04).

[0150] In this experimental group, it was found that the number of hairs that fell out, even when force was applied, continued to decrease significantly from week 0 to week 12. In particular, compared to the control group, the number was significantly lower at 56.3% at week 12. In other words, even after subtracting the placebo effect, daily intake of banana flower extract significantly reduced the number of hairs that fell out, even when pulling forcefully, and improved the stability of hair follicles. The health benefits of hair can be achieved by making each individual hair healthy.

[0151] As described above, any embodiment of the banana flower extract can be used to prepare a hair health care composition. Any embodiment of the banana flower extract of the present invention can increase the diameter of hair roots, improve the stability of hair follicles, reduce hair loss, inhibit the production of dihydrotestosterone (DHT), promote hair growth, and increase the proliferation of hair follicle cells.

[0152] Although the technical contents of the present invention have been described above with reference to preferred embodiments, the present invention is not limited to these embodiments, and all slight changes and modifications that can be made by a person skilled in the art without departing from the spirit of the present invention are included in the scope of the present invention. The scope of protection of the present invention is determined based on the claims.

Claims

1. A hair health care composition comprising an effective amount of banana flower extract and a carrier, wherein the banana flower extract is a compound I represented by formula (I) below, a compound II represented by formula (II) below, or a combination thereof. 【Chemistry 1】 A hair care composition comprising:

2. Compound I represented by the following formula (I), compound II represented by the following formula (II), or a combination thereof 【Chemistry 2】 A banana flower extract characterized by:

3. The banana flower extract is used to prepare a health care composition, Obtained by extracting the stamens of banana flowers through an aqueous extraction step. Use of banana flower extract.

4. The banana flower extract has the effect of increasing the diameter of hair roots.

4. Use of the banana flower extract according to claim 3.

5. The banana flower extract has the effect of increasing the stability of hair follicles.

4. Use of the banana flower extract according to claim 3.

6. The banana extract has the effect of reducing hair loss.

4. Use of the banana flower extract according to claim 3.

7. The banana extract is extracted from the banana flower stamens through an aqueous extraction step, a re-extraction step, and a washing step.

4. Use of the banana flower extract according to claim 3.

8. The banana extract is a compound I represented by the following formula (I), a compound II represented by the following formula (II), or a combination thereof. 【Transformation 3】 8. Use of the banana flower extract according to claim 7.

9. The banana extract has the effect of inhibiting the production of dihydrotestosterone.

8. Use of the banana flower extract according to claim 7.

10. The banana extract has the effect of promoting hair growth.

8. Use of the banana flower extract according to claim 7.

11. The banana extract has the effect of increasing the proliferation of hair follicle cells.

8. Use of the banana flower extract according to claim 7.

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