Compund for hair care composition and use thereof

Abstract

A compound for preparing a hair care composition, where in the compoundis as represented by formula (I), formula (II) ,or a combination thereof. ；and

Description

Technical Field

[0001] This invention relates to a compound and its use, particularly its use in the preparation of compositions for hair care. Prior Technology

[0002] Bananas are a very popular tropical fruit, enjoyed fresh or used in various desserts. Taiwan currently cultivates bananas on approximately 10,000 hectares, producing abundant fruit that meets both domestic and export demands. Banana flowers consist of the male flower at the very tip, as well as neuter and female flowers. Only the ovary at the base of the female flower develops into a fruit; the male flowers are generally discarded to avoid competing for nutrients with the banana fruit. These discarded male flowers are called banana flowers.

[0003] Male pattern baldness is a common cause of hair loss, related to genetics and hormones. The male hormone testosterone in the human body is converted into dihydrotestosterone (DHT) by reductase. Hair is affected by DHT, which causes it to gradually become thinner and fall out. Over time, even the hair follicles will shrink.

[0004] While common treatments like rotopelliptic can prevent the worsening of male pattern baldness, they also have side effects such as decreased libido, erectile dysfunction, and infertility. Summary of the Invention

[0005] In view of this, in order to find a treatment method without side effects and to develop other applications for discarded banana flowers, a hair care composition and compound and its use in the preparation of the hair care composition are proposed.

[0006] In some embodiments, a compound as shown in formula (I):

[0007] Formula (I).

[0008] Here, compound I is N-β-Citroyldopamine, and compound II is 6,2',3',6'-O-tetraacetyl-3-O-trans-p-coumaroylsucrose.

[0009] In some embodiments, a compound as shown in formula (II):

[0010] Equation (II).

[0011] In some embodiments, the compounds of formula (I) and formula (II) are isolated from banana flower extract, which is obtained by extracting banana flower stamens through a water extraction step.

[0012] In some embodiments, the use of the compound for preparing a hair care composition, the compound being a compound of formula (I), formula (II), or a combination thereof.

[0013] In some embodiments, the compound has the effect of inhibiting the production of dihydrotestosterone (DHT).

[0014] In some embodiments, the compound has a hair growth promoting effect.

[0015] In some embodiments, the compound has the effect of increasing hair follicle cell proliferation.

[0016] In summary, the compounds of any of the embodiments can be used to prepare hair care compositions. In some embodiments, the compounds may have at least one of the following effects: inhibiting dihydrotestosterone (DHT) production, promoting hair growth, and increasing hair follicle cell proliferation. Simple Explanation of the Diagram

[0017] Figure 1 shows the results of the inhibition of dihydrotestosterone content by banana flower extract in one embodiment. Figure 2 shows the results of banana flower extract promoting hair follicle cell proliferation in one embodiment. Figure 3 shows the results of banana flower extract promoting hair follicle cell proliferation in another embodiment. Figure 4 shows the results of the relative average hair root diameter in human testing. Figure 5 shows the results of hair follicle stability testing on human subjects. Figure 6 shows the results of the relative average hair loss in human trials. Implementation

[0018] As used in this article, "banana flower" refers to the flower of the banana plant.

[0019] In some embodiments, banana flower extract may be extracted from the banana flowers of banana (Musa sapientum L.), banana flowers of Luzon banana (Musa spp. AAB Silk), banana flowers of pink banana (Musa spp. ABB Bluggoe), banana flowers of canna (Musa spp. AAA Robusta), banana flowers of honey banana (Musa spp. AAB Bluggoe), or banana flowers of banana (Musa paradisiacal).

[0020] In some embodiments, the banana flowers used to extract banana flower extract may be the stamens of banana flowers. The banana flowers may be fresh stamens, dried stamens, or frozen stamens.

[0021] In some embodiments, the banana flowers used to extract banana flower extract may be whole or banana flowers that have undergone physical processing procedures such as chopping, dicing, grinding, or other methods to change the size and integrity of the original material.

[0022] In some embodiments, the extraction step of banana flower extract includes a water extraction step. In some embodiments, the extraction step of banana flower extract further includes a re-extraction step and a rinsing extraction step.

[0023] Water extraction step: Extract banana flower stamens with water to obtain banana flower water extract.

[0024] Re-extraction step: The banana flower aqueous extract was subjected to liquid phase partition extraction in water and n-butanol as solvents to obtain the n-butanol separation layer.

[0025] Rinsing step: The n-butanol separation layer is rinsed separately with water and a methanol solution of mixed water and methanol.

[0026] In some embodiments, the water extraction step involves heating water to a fixed temperature range, adding banana flowers to the water, mixing, and maintaining the temperature for a fixed period of time to obtain a banana flower water extract. In some embodiments, the fixed temperature range refers to 30°C to 65°C, 40°C to 60°C, or 45°C to 55°C. In some embodiments, the fixed period of time refers to 50 to 100 minutes. For example, banana flowers are mixed with water at 50°C and extracted at 50°C for 50 minutes to obtain a banana flower water extract.

[0027] In some embodiments, during the water extraction step, the weight ratio of water to banana flowers is 2~11:1~4. For example, the water:banana flower ratio is 4:1.

[0028] In some embodiments, during the water extraction step, the initial extract obtained by water extraction of banana flower stamens can be used to obtain a banana flower aqueous extract via a centrifugation step and / or a filtration step. The centrifugation step involves centrifuging the initial extract to obtain its supernatant. In some embodiments, the filtration step involves passing the supernatant (or initial extract) obtained from the centrifugation step through a sieve to remove solids from the solution to obtain a filtrate. For example, the sieve can be a 350-mesh sieve. In other words, the banana flower aqueous extract can be the aforementioned initial extract, the supernatant obtained from the centrifugation step, or the filtrate obtained from the filtration step.

[0029] In some embodiments, the ratio of n-butanol to water in the re-extraction step may be 3:1.

[0030] In some embodiments, the extraction step may be a two-stage extraction. The first extraction may be a bioactive guided separation of the n-butanol separation layer with a third-order linear gradient consisting of water and methanol to obtain a pure water separation section and a methanol-water solution separation section. The second extraction may be a reverse-phase-fast column chromatography of the water separation section with a linear extraction gradient from pure water to 50% methanol solution, and a reverse-phase-fast column chromatography of the methanol-water solution separation section with a linear extraction gradient from pure water to 100% methanol.

[0031] In some embodiments, the banana flower extract is compound I as shown in formula (I), compound II as shown in formula (II), or a combination thereof:

[0032] Equation (I); and

[0033] Equation (II).

[0034] Compound I is N-β-Citroyldopamine, while compound II is 6,2',3',6'-O-tetraacetyl-3-O-trans-p-coumaroylsucrose.

[0035] In some embodiments, the banana flower extract may be an aqueous extract of banana flowers, compound I, compound II, or a combination of compound I and compound II.

[0036] In some embodiments, the effective dosage of banana flower extract is 1 gram per day.

[0037] In some embodiments, banana flower extract can be used to prepare hair care compositions.

[0038] In some embodiments, the aforementioned composition may be a health supplement. In other words, this health supplement contains an effective amount of banana flower extract.

[0039] In some embodiments, the aforementioned health composition may be manufactured into a dosage form suitable for enteral or oral administration using techniques known to those skilled in the art. These dosage forms include, but are not limited to: tablets, troche, lozenges, pills, capsules, dispersible powders or granules, solutions, suspensions, emulsions, syrups, elixirs, slurries, and the like.

[0040] In some embodiments, the aforementioned health care composition may be manufactured using techniques well known to those skilled in the art into a dosage form suitable for parenterally or topically administration, including, but not limited to, injections, sterile powders, external preparations, and the like. In some embodiments, the health care composition may be administered via a parenteral route selected from the group consisting of: subcutaneous injection, intraepidermal injection, intradermal injection, and intralesional injection.

[0041] In some embodiments, the health composition may further comprise a food-acceptable carrier widely used in food manufacturing techniques. For example, a food-acceptable carrier may comprise 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. The selection and quantity of these agents fall within the scope of professional competence and routine technique of a person skilled in this art.

[0042] In some embodiments, a 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.

[0043] In some embodiments, the aforementioned health-promoting composition may be an edible composition. In some embodiments, this edible composition may be formulated into a food product or may be a food additive, meaning that it is added during the preparation of ingredients by conventional methods to obtain a food product, or added during the production of a food product. Here, the food product may be a product formulated with edible materials for human or animal consumption.

[0044] In some embodiments, food products may be, but are not limited to: beverages, fermented foods, bakery products, health foods, and dietary supplements.

[0045] [Example 1: Preparation of Banana Flower Extract]

[0046] First, bananas (scientific name: Musa paradisiacal) produced in Taiwan are used, and their stamens are taken as banana flowers.

[0047] Next, using water as a solvent, the water was heated to 50±5℃, and then banana flowers were added. The ratio of banana flowers to water by weight was 1:4. After mixing the banana flowers and water, extraction was maintained at 50±5℃ for 50 minutes to obtain the initial extract.

[0048] Subsequently, the initial extract is sieved through a 350-mesh sieve and the filter is collected to obtain the banana flower aqueous extract.

[0049] Next, 10 liters of the above banana flower aqueous extract were taken and subjected to liquid phase separation extraction using a 3:1 ratio of n-butanol and water to obtain an n-butanol layer extract and an aqueous layer extract. Here, the highly polar substances in the banana flower aqueous extract remain in the n-butanol layer extract, while the water-soluble substances remain in the water layer extract.

[0050] Next, the n-butanol layer extract was concentrated and dried under reduced pressure to obtain 24.6 g of n-butanol layer extract (BUF). The aqueous layer extract was concentrated and dried under reduced pressure to obtain 198.7 g of aqueous layer extract (WF).

[0051] Based on this, it can be calculated that 223.3 grams of powdered extract can be obtained from 10 liters (L) of banana flower extract, of which 11% is n-butanol layer extract (BUF) and 89% is water layer extract (WF).

[0052] Next, using bioassay guided fractionation, 20 g of n-butanol extract was subjected to macroporous resin column chromatography (Diaion HP-20 column chromatography 60-5 cm). Pure water, methanol solution (methanol / pure water = 50 / 50), and methanol (100%) were used as the extraction solvent to sequentially extract three fractions: BU1, BU2, and BU3.

[0053] In the BU1 separation section, reverse-phase rapid column chromatography (RP-MPLC) was used for separation. Extraction was performed sequentially with water to 50% methanol solution (the extraction gradient was a linear gradient from pure water for 0 min to 50% methanol solution for 80 min). Subsequent thin-layer chromatography (TLC) was used, detecting wavelengths of 210 and 280 nm at a flow rate of 20 mL / min. Each 30 mL sample was collected into a separate collection section, resulting in 54 collection sections. Extracts with similar results were combined to obtain five sub-separation sections. These five sub-separation sections were designated BU1-1, BU1-2, BU1-3, BU1-4, and BU1-5.

[0054] The BU1-3 separation fractions were then purified by HPLC (detection wavelength 210 nm), followed by purification using a reverse-phase C-18 column with 20% methanol as the mobile phase to obtain compound I. Compound I was confirmed to be nitrogen-β-citroyldopamine with the following chemical structure (I) by 1H, 13C, 2D-NMR (including HSQC, HMBC, COSY, NOESY) and HR-LC-ESI-MSMS (measured approximate molecular weight m / z 350.0845 [M+Na]+, theoretical approximate molecular formula C14H16NO8, theoretical approximate molecular weight m / z 350.0840, actual molecular formula C14H17NO8).

[0055] Formula (I).

[0056] The BU2 separation section was separated using reverse-phase-rapid column chromatography (RP-MPLC), with sequential extraction from water to methanol (a linear gradient from pure water for 0 min to 100% methanol for 120 min). Thin-layer chromatography was then used, with detection wavelengths of 210 nm and 280 nm at a flow rate of 20 mL / min. Each 30 mL sample was collected, resulting in 80 collection sections. Extracts with similar results were combined to obtain 10 sub-separation sections: BU2-1, BU2-2, BU2-3, BU2-4, BU2-5, BU2-6, BU2-7, BU2-8, BU2-9, and BU2-10.

[0057] The BU2-8 fraction was then purified by HPLC (detection wavelength 310 nm) and purified by reverse-phase C-18 column with 45% methanol as the mobile phase to obtain compound II. Compound II was confirmed to be 6,2',3',6'-oxo-tetraacetyl-3-oxo-trans-p-coumaroylsucrose with the following chemical structure (II) by 1H, 13C, 2D-NMR and HR-LC-ESI-MSMS (measured approximate molecular weight m / z 655.1888 [M+H]+, theoretical approximate molecular formula C29H35O17, theoretical approximate molecular weight m / z 655.1880, actual molecular formula C29H36O17).

[0058] Equation (II).

[0059] Therefore, the banana flower aqueous extract, compound I, and compound II are the banana flower extract.

[0060] [Example 2: Test for the inhibition of dihydrotestosterone by banana flower extract]

[0061] A significant increase in dihydrotestosterone (DHT) production in frontal hair follicles has been observed in young male patients with alopecia. Inhibiting DHT may have the effect of nourishing hair follicles and promoting hair growth. In this test, the DHT content in human prostate cells treated with different concentrations of banana extract was measured to see if it decreased, with a control group using the common androgenetic alopecia treatment drug, rotopelix.

[0062] Materials and Equipment Specifications:

[0063] Cell line: Human prostate cells LNcap (purchased from BCRC; preservation number Cat. 60088), hereinafter referred to as LNcap cells.

[0064] 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).

[0065] Reagents: 1X DPBS (Gibco, Cat. 14200-075), trypsin (1X Trypsin-EDTA, Thermo, Cat. FNN0011), finasteride (Sigma; Cat. F1293), testosterone (Sigma; Cat. T1500), cell lysis buffer (RIPA Lysis and Extraction Buffer (Thermo #89900)).

[0066] Test kit: Dihydrotestosterone ELISA reagent test kit (USCN; CEA443Ge).

[0067] Test process:

[0068] First, LNcap cells were seeded into 6-well cell culture dishes at a density of 1:10⁵ cells per well and cultured at 37°C in a 5% CO₂ incubator for 24 hours.

[0069] 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, experimental group B was supplemented with 5 µg / mL of compound I, control group was supplemented with 20 µg / mL of rotoprene, and the blank group was supplemented with only cell culture medium.

[0070] Next, 10 mg / mL of testosterone was added to each group, and then the mixture was placed in a carbon dioxide incubator at 37°C for 2 hours.

[0071] After removing the supernatant from each group, the culture plates were washed twice with 1XDPBS, 200µL of trypsin was added, and the LNcap cells were incubated in a 37°C, 5% CO2 incubator for 3 minutes. The cell culture plates were then gently tapped to suspend the LNcap cells.

[0072] Subsequently, 600 µL of cell culture medium was added to each well to stop the action of trypsin. The cell solution from each well was then collected and placed into a 1.5 mL microcentrifuge tube, and the microcentrifuge tube was centrifuged at 400 xg for 5 minutes. After removing the supernatant from the microcentrifuge tube, 200 μL of 1X DPBS was added to the tube to wash the LNcap cells, and the microcentrifuge tube was centrifuged again for 5 minutes before removing the supernatant.

[0073] Next, add 200 µL of 1X cell lysis buffer to the tube to lyse LNcap cells. Centrifuge at 13,000 rpm for 5 minutes at 4°C, and collect the supernatant in a 1.5 mL microcentrifuge tube. Finally, use the dihydrotestosterone ELISA kit to detect the dihydrotestosterone content in LNcap cells. (Follow the manufacturer's instruction manual for detailed procedures.)

[0074] Test results:

[0075] Therefore, the two groups were compared using a one-tailed Student's t-test to determine whether there was a statistically significant difference, and to obtain their p-values. In the figure, "*" represents a p-value less than 0.05, and "**" represents a p-value less than 0.01. More "*" characters indicate a more significant statistical difference.

[0076] Please refer to Figure 1. The dihydrotestosterone (DHT) content in the blank group was 32.8 pg / mL, the DHT content in the control group was 17.16 pg / mL, the DHT content in experimental group A was 25.03 pg / mL, and the DHT content in experimental group B was 25.20 pg / mL. It can be seen that, regardless of whether the product used was Rovell or banana extract, experimental groups A and B showed statistically significant differences compared to the blank group, indicating an inhibitory effect on DHT.

[0077] Furthermore, banana extract B at a concentration of 5 µg / mL showed a significant difference in efficacy compared to the control group, and at a concentration of 10 µg / mL, banana extract B exhibited the same significant efficacy (p value less than 0.01) as the commercially available drug, Rospri. Therefore, it can be concluded that banana flower extract has uses for hair health.

[0078] [Example 3: Test on the effect of banana flower extract on hair follicle cell proliferation]

[0079] Hair follicle cells are the foundation of hair. During the growth phase, the cells within the hair follicle bulb at the hair root can divide once a day. Promoting the proliferation of hair follicle cells can promote hair growth. In this test, the proliferation of human hair follicle cells treated with compound I was investigated.

[0080] Materials and Equipment Specifications:

[0081] Cell line: Human Follicle Dermal Papilla Cells (HFDPC) (purchased from PromoCell; Cat. C-12071), hereinafter referred to as HFDPC cells.

[0082] Cell culture medium: Follicle Dermal Papilla Cell Growth Medium (purchased from PromoCell; Cat. C-26501).

[0083] Reagents: 10X DPBS (Gibco, Cat. 14200-075), trypsin (1X Trypsin-EDTA, Thermo, Cat. FNN0011).

[0084] Detection kit: Cell proliferation ELISA reagent detection kit (Click-iT™ Plus EdU Flow Cytometry Assay Kits - Alexa Fluor™ 488 picolyl azide, 50 tests, Invitrogen, model C10632), containing composition C (Dimethyl sulfoxide DMSO), composition D (Click-iT™ fixative), composition E (Click-iT™ saponin-based permeabilization and wash reagent), composition F (Copper protectant), and composition G (Click-iT™ EdU buffer additive).

[0085] Test process:

[0086] First, HFDPC cells were seeded into 6-well cell culture dishes at a density of 1:10⁵ cells / well and cultured at 37°C in a 5% CO₂ incubator for 24 hours.

[0087] The cultured HFDPC cells were divided into an experimental group and a control group. The experimental group was supplemented with 10 µg / mL of compound I prepared in Example 1, while the control group was supplemented with only cell culture medium.

[0088] Next, 10 μM of composition C was added to each group, and the cells were incubated in a CO2 incubator at 37°C for 2 hours. After removing the cell culture medium from each group, the culture plates were washed once with 1XDPBS, 200 µL of trypsin was added, and the HFDPC cells were incubated in a 5% CO2 incubator at 37°C for 5 minutes. The cell culture plates were then gently tapped to suspend the HFDPC cells.

[0089] Subsequently, after removing the supernatant, 200 µL of cell culture medium was added to each well to stop the action of trypsin. The cell solution from each well was then collected and placed into a 1.5 mL microcentrifuge tube, and the microcentrifuge tube was centrifuged at 400 xg for 5 minutes. After removing the supernatant from the microcentrifuge tube, it was washed with 1XDPBS, centrifuged (400 xg for 5 minutes), and after removing the supernatant, 100 μL of composition D was added to the tube and it was incubated at room temperature in the dark for 10 minutes.

[0090] Next, after removing the supernatant, wash again with 1XDPBS, centrifuge (400 xg for 5 minutes), remove the supernatant, add 100 μL of composition E to the tube, and incubate at room temperature in the dark for 15 minutes. Then add 300 μL of 1% BSA / PBS wash buffer and centrifuge at 400 xg for 5 minutes (this step is repeated three times).

[0091] After removing the supernatant, add 100 μL of composition E to each tube and incubate at room temperature in the dark for 15 minutes. Add 100 μL of the prepared Click-iT™ reaction mixture (prepared according to the Cell Proliferation ELISA Reagent Detection Kit Instructions) to each centrifuge tube and incubate at room temperature in the dark for 30 minutes. Add 300 μL of 1% BSA / PBS wash buffer and centrifuge at 400xg for 5 minutes. Finally, measure the fluorescence signal using flow cytometry (excitation light: 488 nm; scattered light: 527 nm & 590 nm).

[0092] Test results:

[0093] Therefore, the two groups were compared using a one-tailed Student's t-test to determine whether there was a statistically significant difference, and to obtain their p-values. In the figure, "*" represents a p-value less than 0.05, and "**" represents a p-value less than 0.01. More "*" characters indicate a more significant statistical difference.

[0094] Please refer to Figure 2. If the number of HFDPC cells in the control group is considered 100%, then the calculated number of HFDPC cells in the experimental group is 118.81%. This indicates that compound I has a statistically significant difference compared to the control group, meaning that banana flower extract promotes hair follicle cell proliferation.

[0095] [Example 4: Test on the effect of banana flower extract on hair follicle cell proliferation]

[0096] Hair follicle cells are the foundation of hair. During the growth phase, the cells within the hair follicle bulb at the hair root can divide once a day. Promoting the proliferation of hair follicle cells can promote hair growth. In this test, the proliferation of human hair follicle cells treated with compound II was observed.

[0097] Materials and Equipment Specifications:

[0098] Cell line: Human Follicle Dermal Papilla Cells (HFDPC) (purchased from PromoCell; Cat. C-12071), hereinafter referred to as HFDPC cells.

[0099] Cell culture medium: Follicle Dermal Papilla Cell Growth Medium (purchased from PromoCell; Cat. C-26501).

[0100] Reagents: 10X DPBS (Gibco, Cat. 14200-075), trypsin (1X Trypsin-EDTA, Thermo, Cat. FNN0011).

[0101] Detection kit: Cell proliferation ELISA reagent detection kit (Click-iT™ Plus EdU Flow Cytometry Assay Kits - Alexa Fluor™ 488 picolyl azide, 50 tests, Invitrogen, model C10632), containing composition C (Dimethyl sulfoxide DMSO), composition D (Click-iT™ fixative), composition E (Click-iT™ saponin-based permeabilization and wash reagent), composition F (Copper protectant), and composition G (Click-iT™ EdU buffer additive).

[0102] Test process:

[0103] First, HFDPC cells were seeded into 6-well cell culture dishes at a density of 1:10⁵ cells / well and cultured at 37°C in a 5% CO₂ incubator for 24 hours.

[0104] The cultured HFDPC cells were divided into an experimental group, a (positive) control group, and a blank group. The experimental group was supplemented with 100 μM of compound II prepared in Example 1, the control group was supplemented with 20% FBS (fetal bovine blood), and the blank group was supplemented with only cell culture medium.

[0105] Next, 10 μM of composition C was added to each group, and the cells were incubated in a CO2 incubator at 37°C for 2 hours. After removing the cell culture medium from each group, the culture plates were washed once with 1XDPBS, 200 µL of trypsin was added, and the HFDPC cells were incubated in a 5% CO2 incubator at 37°C for 5 minutes. The cell culture plates were then gently tapped to suspend the HFDPC cells.

[0106] Subsequently, after removing the supernatant, 200 µL of cell culture medium was added to each well to stop the action of trypsin. The cell solution from each well was then collected and placed into a 1.5 mL microcentrifuge tube, and the microcentrifuge tube was centrifuged at 400 xg for 5 minutes. After removing the supernatant from the microcentrifuge tube, it was washed with 1XDPBS, centrifuged (400 xg for 5 minutes), and after removing the supernatant, 100 μL of composition D was added to the tube and it was incubated at room temperature in the dark for 10 minutes.

[0107] Next, after removing the supernatant, wash again with 1XDPBS, centrifuge (400 xg for 5 minutes), remove the supernatant, add 100 μL of composition E to the tube, and incubate at room temperature in the dark for 15 minutes. Then add 300 μL of 1% BSA / PBS wash buffer and centrifuge at 400 xg for 5 minutes (this step is repeated three times).

[0108] After removing the supernatant, add 100 μL of composition E to each tube and incubate at room temperature in the dark for 15 minutes. Add 100 μL of the prepared Click-iT™ reaction mixture (prepared according to the Cell Proliferation ELISA Reagent Detection Kit Instructions) to each centrifuge tube and incubate at room temperature in the dark for 30 minutes. Add 300 μL of 1% BSA / PBS wash buffer and centrifuge at 400xg for 5 minutes. Finally, measure the fluorescence signal using flow cytometry (excitation light: 488 nm; scattered light: 527 nm & 590 nm).

[0109] Test results:

[0110] Therefore, the two groups were compared using a one-tailed Student's t-test to determine whether there was a statistically significant difference, and to obtain their p-values. In the figure, "*" represents a p-value less than 0.05, and "**" represents a p-value less than 0.01. More "*" characters indicate a more significant statistical difference.

[0111] Please refer to Figure 3. Taking the number of HFDPC cells in the blank group as 100%, the calculated number of HFDPC cells in the control group is 117.01%, and the number of HFDPC cells in the experimental group is 112.54%. This shows that compound II has a statistically significant difference compared to the blank group, meaning that banana flower extract promotes hair follicle cell proliferation.

[0112] [Example 5: Human testing of banana flower extract]

[0113] Participants: 50 participants (divided into a control group of 25 participants and an experimental group of 25 participants). All participants were adults aged 20 or older.

[0114] Test items: hair root diameter, hair follicle stability, and average hair loss during shampooing.

[0115] The hairline diameter was measured using a digital micrometer, model Mitutoyo C / N293-100.

[0116] Hair follicle stability refers to applying force to pull hairs (approximately 60 hairs per area) upwards from three regions: 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, the better the hair follicle stability.

[0117] The average amount of hair loss during shampooing refers to the amount of hair loss observed during shampooing.

[0118] Testing method:

[0119] Twenty-five participants in the experimental group consumed a powder made from dried banana flower extract prepared in Example 1, containing 1 gram of the extract daily, while another 25 participants in the control group consumed a placebo daily without banana flower extract, for 12 consecutive weeks.

[0120] The data measured before consumption (i.e., week 0) are referred to as control group 01 and experimental group 01, respectively. The data measured after 4 weeks of consumption (i.e., week 4) are referred to as control group 02 and experimental group 02, respectively. The data measured after 8 weeks of consumption (i.e., week 8) are referred to as control group 03 and experimental group 03, respectively. The data measured after 12 weeks of consumption (i.e., week 12) are referred to as control group 04 and experimental group 04, respectively.

[0121] Test results:

[0122] The graph below shows the relative values ​​for each group, calculated using the average of all subjects and taking week 0 as 100%. The standard deviation was calculated using the STDEV formula in Excel, and a one-tailed Student's t-test was used in Excel to analyze whether there were statistically significant differences, obtaining the p-value. In the graph, "*" indicates a p-value less than 0.05, "**" indicates a p-value less than 0.01, and "***" indicates a p-value less than 0.001, representing a statistically significant difference. A control group with the same number of weeks was also compared, and a t-test was used to analyze whether there were statistically significant differences, obtaining the p-value. In the graph, "#" indicates a p-value less than 0.05, "##" indicates a p-value less than 0.01, and "###" indicates a p-value less than 0.001, representing a statistically significant difference.

[0123] Please refer to Figure 4. After 4 weeks of daily administration of banana flower extract, the average hair root diameter of the 25 subjects in the experimental group increased from 100% (experimental group 01) in week 0 to 102.5% (experimental group 02). After continuing daily administration of banana flower extract to week 8, the average hair root diameter increased to 103.2% (experimental group 03). After continuing daily administration of banana flower extract to week 12, the average hair root diameter significantly increased to 107.8% (experimental group 04).

[0124] See Figure 4. The average hair root diameter in the placebo group increased from 100% (control group 01) in week 0 to 101.3% (control group 02). After continuing to take the placebo daily until week 8, the average hair root diameter increased to 102.4% (control group 03). However, after continuing to take the placebo daily until week 12, the average hair root diameter decreased to 101.4% (control group 04).

[0125] Generally, thinning hair is one of the initial symptoms of male pattern baldness. In other words, when the diameter of the hair root is measured to be increasingly larger, it indicates that the male pattern baldness is beginning to improve. In this test, the experimental group showed an increasing hair root diameter from week 0 to week 12, with a significant 6.4% increase compared to the control group at week 12. This means that even after adjusting for the placebo effect, daily consumption of banana flower extract can significantly increase hair root diameter, resulting in healthier hair and thus a hair care effect.

[0126] Please refer to Figure 5. After 4 weeks of daily administration of banana flower extract, the hair follicle stability of the 25 subjects in the experimental group decreased from 100% in week 0 (experimental group 01) to 86.7% (experimental group 02). After continuing daily administration of banana flower extract to week 8, the hair follicle stability decreased to 35.6% (experimental group 03). After continuing daily administration of banana flower extract to week 12, the hair follicle stability decreased significantly to 17.8% (experimental group 04).

[0127] See Figure 5. The relative hair loss in the placebo group decreased from 100% (control group 01) in week 0 to 88.6% (control group 02). After continuing to take the placebo daily, the average hair root diameter decreased to 85.7% in week 8 (control group 03). After continuing to take the placebo daily, the relative hair loss remained at 85.7% in week 12 (control group 04).

[0128] In this test, the experimental group showed a consistently significant reduction in hair loss even when pulled, from week 0 to week 12, especially compared to the control group at week 12, where the loss was significantly lower by 67.9%. This means that even after adjusting for the placebo effect, daily consumption of banana flower extract can significantly reduce hair loss when pulled, achieving better hair follicle stability and resulting in healthier hair strands and overall hair health benefits.

[0129] Please refer to Figure 6. After 12 weeks of daily administration of banana flower extract, the relative average hair loss during shampooing in the 25 subjects in the experimental group significantly decreased from 100% (experimental group 01) at week 0 to 66.9% (experimental group 04). Continue to refer to Figure 6. In contrast, the relative average hair loss in the placebo group increased from 100% (control group 01) at week 0 to 123.2% (control group 04).

[0130] In this test, the experimental group showed a consistently significant reduction in hair loss even when pulled, from week 0 to week 12, especially compared to the control group at week 12, where the loss was significantly lower by 56.3%. This means that even after adjusting for the placebo effect, daily consumption of banana flower extract can significantly reduce the amount of hair lost when pulled, achieving better hair follicle stability and resulting in healthier hair for each individual hair, thus achieving a hair care effect.

[0131] In summary, the banana flower extract of any embodiment can be used to prepare a hair care composition. The banana flower extract of any embodiment can be used to increase hair root diameter, improve hair follicle stability, reduce hair loss, inhibit dihydrotestosterone (DHT) production, promote hair growth, and enhance hair follicle cell proliferation.

[0132] Although the technical content of the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the present invention. Any modifications and refinements made by those skilled in the art without departing from the spirit of the present invention should be included within the scope of the present invention. Therefore, the scope of protection of the present invention shall be determined by the appended claims.

[0133] none

Claims

1. A compound as shown in formula (I): Formula (I).

2. The compound as claimed in claim 1, wherein the compound is isolated from banana flower extract.

3. A compound as shown in formula (II): Formula (II).

4. The compound as claimed in claim 3, wherein the compound is isolated from banana flower extract.

5. Use of a compound for preparing a hair care composition, the compound being a compound of formula (I), a compound of formula (II), or a combination thereof: formula (I); and formula (II).

6. The use as described in claim 5, wherein the compound is used to inhibit the production of dihydrotestosterone.

7. The use as described in claim 5, wherein the compound is used to promote hair growth.

8. The use as described in claim 7, wherein the compound is used to promote hair follicle cell proliferation.

Images