Traditional Chinese medicine composition for preventing hair loss and growing hair by inhibiting DHT receptor expression and preparation method thereof
The nanocomposite formed by the self-assembly of noni fruit extract FEA-3 and molybdate inhibits androgen receptor expression, solving the problem that finasteride cannot block androgen receptors, and achieving a safe and effective effect in preventing hair loss and promoting hair growth.
Patent Information
- Application Number
- CN202511617707.1
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-11-06
- Publication Date
- 2026-01-02
AI Technical Summary
In existing technologies, although the 5α-reductase inhibitor finasteride can reduce DHT concentration, it cannot block androgen receptors that have been occupied by DHT, leading to recurrence of hair loss. Furthermore, it has side effects due to systemic distribution and cannot achieve safe and long-term topical inhibition of hair follicle-specific androgen receptors.
A nanocomposite was formed by the self-assembly of noni fruit extract FEA-3 and molybdate. By inhibiting the expression of androgen receptors, noni fruit extract FEA-3 specifically binds to androgen receptors and promotes their degradation. At the same time, molybdenum-based nanoparticles block downstream signaling pathways of androgen receptors in cells, thereby achieving hair follicle miniaturization and promoting hair growth.
It achieves dual inhibition of androgen receptors within hair follicles, degrades androgen receptor proteins and downregulates gene transcription, promotes hair growth, avoids systemic side effects, and provides a safe and reliable effect in preventing hair loss and promoting hair growth.
Abstract
Description
Technical Field
[0001] This invention relates to the field of hair growth and hair care products, specifically to a traditional Chinese medicine composition that prevents hair loss and promotes hair growth by inhibiting DHT receptor expression, and its preparation method. Background Technology
[0002] Hair follicles are the only organs in the human body capable of periodic regeneration, and their health directly determines the quality of healthy hair growth. However, with age or increased androgen levels, DHT (dihydrotestosterone) accumulates around hair follicles, leading to a decline in the activity of hair follicle cells.
[0003] The dihydrotestosterone (DHT) receptor is the androgen receptor. Androgen receptors are nuclear receptors that bind to androgens (such as testosterone and dihydrotestosterone), thereby regulating gene expression and cellular function.
[0004] The 5α-reductase inhibitor finasteride only reduces DHT concentration but cannot block androgen receptors already occupied by DHT; DHT levels rebound after discontinuation, and hair loss recurs. Because androgen receptors are widely distributed throughout the body, finasteride causes side effects such as gynecomastia and liver toxicity, and topical application of finasteride is also difficult to accumulate in the hair follicle microenvironment. Therefore, there is an urgent need in this field for a composition that is "specific to hair follicles, downregulates androgen receptors, and is safe for long-term topical application." Summary of the Invention
[0005] In order to overcome the shortcomings of the prior art, one of the objectives of this invention is to provide a traditional Chinese medicine composition that achieves hair loss prevention and hair growth by inhibiting the expression of DHT receptors. This traditional Chinese medicine composition achieves the degradation of androgen receptor protein and the silencing of androgen receptor gene in hair follicle cells, breaks through the bottleneck of single-target inhibition, and solves the toxic side effects of the chemical antagonist finasteride.
[0006] The second objective of this invention is to provide a method for preparing a traditional Chinese medicine composition that prevents hair loss and promotes hair growth by inhibiting the expression of DHT receptors. This method is simple and has low economic cost.
[0007] The third objective of this invention is to provide a use of a traditional Chinese medicine composition that achieves hair loss prevention and hair growth by inhibiting DHT receptor expression.
[0008] One of the objectives of this invention is achieved through the following technical solution: A traditional Chinese medicine composition for preventing hair loss and promoting hair growth by inhibiting DHT receptor expression includes a nanocomposite. The nanocomposite is formed by coordination-driven self-assembly of noni fruit extract FEA-3 and molybdate. Noni fruit extract FEA-3 acts as a reducing agent and stabilizer for molybdate, inducing the formation of molybdate-based nanoparticles with a particle size of 50-1200 nm. Noni fruit extract FEA-3 is rich in polyphenols and can specifically bind to androgen receptors on the surface of hair follicle cells, achieving targeted delivery of molybdate. The molybdenum-based nanoparticles inhibit androgen receptor nuclear translocation and DHT binding within cells, blocking downstream signaling pathways of androgen receptors, thereby inhibiting hair follicle miniaturization and promoting hair growth.
[0009] Furthermore, the noni fruit extract FEA-3 is prepared by the following method: noni fruit raw material is extracted with ethanol, concentrated, and then extracted with petroleum ether, ethyl acetate and n-butanol in sequence. The ethyl acetate layer is collected and then purified by macroporous resin column chromatography and gel column chromatography to obtain the noni fruit extract FEA-3.
[0010] Furthermore, the molybdenum-based nanoparticles are introduced in the form of molybdate, which is selected from sodium molybdate, ammonium molybdate, or potassium molybdate.
[0011] Furthermore, the nanocomposite has a particle size of 50-1150 nm and a zeta potential of -20 mV to -30 mV.
[0012] Furthermore, the nanocomposite can synergistically inhibit the expression of androgen receptors, wherein: the noni fruit extract FEA-3 promotes the degradation of androgen receptor proteins by binding to the androgen receptor ligand-binding domain; after the molybdenum-based nanoparticles are released into cells, they downregulate the transcriptional level of androgen receptor genes, interfere with androgen receptor transcriptional activity, upregulate SULT-related genes, and downregulate the expression of iNOS, COX2, and Il1f8.
[0013] Furthermore, the herbal composition comprises the aforementioned nanocomposite, an active extract of traditional Chinese medicine for treating androgenetic alopecia, and a pharmaceutically acceptable carrier, and is used for topical treatment of androgenetic alopecia.
[0014] Furthermore, the dosage form of the traditional Chinese medicine composition is a gel or cream.
[0015] The second objective of this invention is achieved by the following technical solution: A method for manufacturing a traditional Chinese medicine composition that prevents hair loss and promotes hair growth by inhibiting DHT receptor expression includes the following steps: S1. The preparation of the nanocomposite includes the following steps: S11. Dissolve the noni fruit extract FEA-3 in the aqueous phase; S12, Add an aqueous solution containing the molybdate; S13. Nanoparticles are formed by self-assembly under ultrasonic or stirring conditions; S14. The nanocomposite was obtained by dialysis, concentration and lyophilization; S2. Compound of active components of traditional Chinese medicine: Take 10-30 parts of Platycladus orientalis leaves, 5-15 parts of Polygonum multiflorum, 5-15 parts of Ligustrum lucidum, and 3-10 parts of fresh ginger. Extract twice by reflux with 50-80% ethanol. Combine the filtrates and recover the ethanol under reduced pressure until there is no alcohol taste. Obtain a fluid extract with a relative density of 1.05-1.15, which is denoted as component B. S3, Composition Molding S31. Matrix preparation: Disperse 0.5-1.5 parts of carbomer 940, 5-10 parts of glycerol, and 0.02-0.1 parts of EDTA-2Na in deionized water at 60-80℃, and neutralize to pH 6.0-7.0 to obtain a transparent gel matrix. S32. Homogenization: 1-5 parts of the nanocomposite, 5-15 parts of component B, 0.1-0.5 parts of menthol, and 0.3-0.8 parts of preservative are added sequentially to the transparent gel matrix, and the mixture is homogenized under vacuum at 6000-12000 rpm for 3-10 min to remove bubbles. S33. Filling: Cool to below 30°C and fill into aluminum-plastic tubes or PET bottles under aseptic conditions to obtain the traditional Chinese medicine composition that achieves hair loss prevention and hair growth by inhibiting DHT receptor expression.
[0016] Compared with the prior art, the beneficial effects of the present invention are as follows: (1) The present invention provides a traditional Chinese medicine composition for preventing hair loss and promoting hair growth by inhibiting the expression of DHT receptors. The nanocomplex inhibits the expression of androgen receptors (androgen receptors) through a dual mechanism: directly by binding androgen receptors through noni fruit extract FEA-3 and promoting their degradation, and indirectly by downregulating the transcription of androgen receptor genes through molybdenum, thereby synergistically exerting the effect of preventing hair loss and promoting hair growth. Noni fruit extract FEA-3 is not only an active ingredient, but also a natural nanocarrier and targeting ligand. It can encapsulate molybdenum ions to form a stable noni fruit-molybdenum nanoparticle complex. Noni fruit extract FEA-3 utilizes its high affinity for androgen receptors to target molybdenum nanoparticles to dermal papilla cells that highly express androgen receptors. After endocytosis, the nanocomplex releases molybdenum ions, activates intracellular antioxidant pathways (such as Nrf2), upregulates the SULT gene, downregulates iNOS / COX2, improves the hair follicle microenvironment, and achieves dual inhibition of androgen receptor expression. Detailed Implementation
[0017] The present invention will now be further described in conjunction with specific embodiments. It should be noted that, without conflict, the embodiments or technical features described below can be arbitrarily combined to form new embodiments.
[0018] Example 1: Preparation of FEA-3 from Noni Fruit Take 10 kg of dried noni fruit, crush it, and extract it three times by refluxing with 70% ethanol for 2 hours each time. Combine the extracts and concentrate them under reduced pressure to obtain an extract. Suspend the extract in water and extract it sequentially with petroleum ether, ethyl acetate, and n-butanol in equal volume ratios. Collect the ethyl acetate layer and elute it with an AB-8 macroporous resin column with a gradient of ethanol from 30% to 70% by volume. Collect the 70% ethanol eluent (FEA) and then purify it by Sephadex LH-20 gel column to obtain the active subfraction FEA-3 (yield 0.8%, HPLC ≥ 92%).
[0019] Noni fruit FEA-3 FE = FruitEthylacetatefraction In Chinese, it is often written as "fruit ethyl acetate extract phase"—that is, the medium polar fraction obtained by further extracting the 70% ethanol extract of noni fruit with ethyl acetate.
[0020] A-3 is the third active subcomponent obtained by eluting with 70% ethanol when passing through an AB-8 macroporous resin column (some laboratories may refer to 30%, 50%, and 70% as A-1, A-2, and A-3, respectively).
[0021] FEA-3 is a "flavonoid-phenolic acid enriched mixture" rather than a single active monomer. Its main active ingredients include rutin, quercetin-3-O-glucoside, kaempferol-3-O-rutin glycoside, etc.
[0022] Example 2: Preparation of Nanocomposites 1. Solution preparation FEA-3 stock solution: Accurately weigh 200 mg (±0.2 mg) of FEA-3 lyophilized powder and place it in a 50 mL low-adsorption polypropylene centrifuge tube. Add 40 mL of ultrapure water (18.2 MΩ·cm, 0.22 μm filtered). Incubate at 60 °C in a water bath and stir magnetically at 1500 rpm (5 cm PTFE stir bar) for 30 min to obtain a clear brownish-red solution (pH 5.8±0.1). Accurately weigh 80 mg (±0.1 mg) of Na2MoO4·2H2O, dissolve it in 10 mL of ultrapure water, filter it through a 0.22 μm aqueous filter membrane, store it at 4 °C protected from light, and use it within 4 hours. 2. Self-assembly: Maintain the FEA-3 solution at 60℃ and 1500 rpm with stirring. Use a 1 mL disposable sterile syringe (0.45 mm needle) to dispense the solution at a rate of 1 mL / min. -1 The molybdate solution was slowly added dropwise at a constant flow rate, protected from light throughout the process. Immediately after addition, the container was transferred to an ice bath and cooled to 25°C within 5 minutes. 3. Ultrasonic shaping Transfer the above mixture into a 50mL glass ultrasonic tube (cylindrical, 2.5cm inner diameter), place it in a CNC ultrasonic cleaner (KQ-300VDE, 35kHz, 300W), and sonicate for 30 minutes in a constant temperature circulating water bath at 25℃ (5s operation / 2s pause, actual acoustic energy 180W). 4. Purification and drying Dialysis: Immediately transfer the opalescent solution into a 100kDa MWCO regenerated cellulose dialysis bag (flat, 24mm wide, 15mm in diameter), clamp and seal both ends, place in 2L of ultrapure water, and magnetically stir at 25℃ and 400rpm. Change the water every 2 hours for a total of 6 times (12 hours).
[0023] Verification of free molybdenum removal rate: Take 1 mL of external solution and determine the Mo concentration by ICP-MS. A Mo concentration of <50 ppb is considered the endpoint of dialysis.
[0024] Concentration: After initial filtration of the dialysate through a 0.45μm needle filter membrane, the solution was concentrated to 5mL using a 3kDa ultrafiltration centrifuge tube (4000g, 4℃, 15min×3).
[0025] Freeze-drying: Pre-freeze at −80℃ for 4 h, then transfer to a freeze dryer (ChristAlpha 1-4, 0.04 mbar, −55℃) and dry for 24 h to obtain 178±6 mg of light brown flocculent powder, yield 89%. The encapsulation efficiency was determined to be 87%, and the molybdenum loading was 11.2%. Example
[0026] This embodiment provides a method for manufacturing a traditional Chinese medicine composition that prevents hair loss and promotes hair growth by inhibiting DHT receptor expression, including the following steps: S1. The preparation of the nanocomposite includes the following steps: S11. Dissolve the noni fruit extract FEA-3 in the aqueous phase; S12, Add an aqueous solution containing the molybdate; S13. Nanoparticles are formed by self-assembly under ultrasonic or stirring conditions; S14. The nanocomposite was obtained by dialysis, concentration and lyophilization; S2. Compound of active components of traditional Chinese medicine: Take 10-30 parts of Platycladus orientalis leaves, 5-15 parts of Polygonum multiflorum, 5-15 parts of Ligustrum lucidum, and 3-10 parts of fresh ginger. Extract twice by reflux with 50-80% ethanol. Combine the filtrates and recover the ethanol under reduced pressure until there is no alcohol taste. Obtain a fluid extract with a relative density of 1.05-1.15, which is denoted as component B. S3, Composition Molding S31. Matrix preparation: Disperse 0.5-1.5 parts of carbomer 940, 5-10 parts of glycerol, and 0.02-0.1 parts of EDTA-2Na in deionized water at 60-80℃, and neutralize to pH 6.0-7.0 to obtain a transparent gel matrix. S32. Homogenization: 1-5 parts of the nanocomposite, 5-15 parts of component B, 0.1-0.5 parts of menthol, and 0.3-0.8 parts of preservative are added sequentially to the transparent gel matrix, and the mixture is homogenized under vacuum at 6000-12000 rpm for 3-10 min to remove bubbles. S33. Filling: Cool to below 30°C and fill into aluminum-plastic tubes or PET bottles under aseptic conditions to obtain the traditional Chinese medicine composition that achieves hair loss prevention and hair growth by inhibiting DHT receptor expression.
[0027] Example 4: In vitro downregulation of androgen receptors Human dermal papilla cells (hDPC, 5 × 10⁻⁶) 4 The nanocomposite was divided into a control group, a DHT model group, and low / medium / high dose groups. The low / medium / high dose groups received 10 µg / mL of nanocomposite. -1 After 24 hours, Western blot results showed that the nanocomposite reduced androgen receptor protein expression in a dose-dependent manner. p <0.01); qPCR results showed that the androgen receptor mRNA level was significantly downregulated, indicating that its action occurs at the transcriptional level; when androgens bind to androgen receptors, the androgen receptors enter the nucleus, bind to the PSA promoter, luciferase is transcribed, and emits a light signal. After treatment with the nanocomplex, the expression level of the PSA-luc reporter gene induced by 10 nM androgens decreased by 72% ( p <0.01 vs model group).
[0028] Example 5: Efficacy evaluation of the nanocomposite FEA-3@Mo in a DHT-induced male pattern baldness model in rabbits. 1. Laboratory animals and adaptive rearing Male New Zealand White rabbits, weighing 2.2±0.2 kg, 12 weeks old, SPF grade, provided by the Experimental Animal Center of Guangdong Medical University. They were housed individually in cages at a temperature of 22±2℃ and a relative humidity of 55±10%, with 12-hour light-dark cycles, and free access to food and water. After 7 days of acclimatization, they were randomly divided into groups of 12 rabbits each. A 6cm×6cm area on their backs was shaved to confirm that the hair follicles were in the telogen phase.
[0029] 2. Model Establishment Starting from day 0, the model group and each drug administration group applied 4 mg / kg of dihydrotestosterone (DHT) oleyl alcohol solution to the shaved area daily. -1 (Equivalent to 0.5 mL, containing 8 mg / mL DHT)-1 The treatment was performed continuously for 28 days; the control group was treated with only an equal amount of oleyl alcohol solvent. Dihydrotestosterone solution was prepared fresh and stored at 4°C protected from light for no more than 72 hours. From day 7 of modeling, visible thinning and decreased hair density appeared in the shaved area, confirming the success of the androgenetic alopecia model.
[0030] 3. Test substance and gel matrix The lyophilized nanocomposite (FEA-3@Mo) powder was prepared according to Example 2, with a molybdenum loading of 11.2%, a particle size of 112±4 nm, a PDI of 0.18, and a ζ-potential of −28.6 mV. It was dried at 4 °C and stored away from light.
[0031] 5% Nanocomposite Gel: Carbomer 940 1.0%, Glycerin 5%, Triethanolamine 1.2%, Phenoxyethanol 0.5%, EDTA-2Na 0.02%, plus ultrapure water to 100%; Nanocomposite 50 mg / g -1 (i.e., 5% w / w), vortex-ultrasound (35 kHz, 5 min) uniformly dispersed, stored in a sealed container at 4 ℃, and used within 7 days.
[0032] 0.1% Finasteride Gel: Finasteride raw material (purity 99.4%) 1 mg / g -1 Prepared with the same substrate, and operated in the dark.
[0033] Blank gel: Contains no active ingredients; all other ingredients are the same as above.
[0034] Apply twice daily at 8:00 AM and 6:00 PM, ensuring the gel evenly covers the shaved area and gently massaging for 30 seconds to ensure no residue remains. Repeat twice daily for four weeks. Before application, wipe away any residual DHT from the previous application with a damp gauze to avoid cross-contamination.
[0035] 4. Macroeconomic Evaluation Image acquisition: Back macro photos were taken on days 0, 7, 14, 21, and 28 using the same light source, the same shooting stand, and a fixed focal length (Canon EOS 90D, 100mm macro lens, color temperature 5500K).
[0036] Hair growth score (Double-blind, two independent observers): 0 points = no new hair growth; 1 point = sparse vellus hair; 2 points = visible short hairs, density <30%; 3 points = density 30-160%; 5 points = complete coverage, density >90%. Intermediate values are allowed in 0.5-point increments.
[0037] Results: The mean value of the blank group was 4.8±0.2; the mean value of the model group was 1.8±0.2; the mean value of the FEA-3@Mo group was 4.5±0.3; and the mean value of the finasteride group was 4.3±0.4 (Mean±SD, n=12). The mean value of the FEA-3@Mo group was significantly higher than that of the model group (P<0.001, One-way ANOVA+Bonferroni).
[0038] 5. Histology and morphometrics Sample collection: On day 29, the patient was euthanized by air embolism in the marginal ear vein. Immediately, a 1.0cm × 1.0cm full-thickness skin sample was taken from the center of the back, fixed in 4% neutral formaldehyde for 24 hours, decalcified with EDTA for 7 days, routinely embedded in paraffin, and serially sectioned in the sagittal plane at 4μm.
[0039] Staining: HE staining results.
[0040] Hair follicle density: Under a 4× objective lens, five non-overlapping fields of view (0.785 mm² / field of view) were randomly selected from each slice, and the total number of hair follicles in the dermis was counted and the average value was taken.
[0041] Results: FEA-3@Mo group: 28.4 ± 2.1 mm -2 Model group 17.5±1.6 mm -2 Increased by 62% p <0.001).
[0042] Anagen phase ratio: determined based on the location of the lower end of the hair follicle (whether the hair bulb is located in the subcutaneous fat layer) and the morphology of the dermal papilla. Count 100 hair follicles per case.
[0043] Results: The FEA-3@Mo group showed a 45 percentage point increase (71±4%), while the model group showed a 26±3% increase (P<0.001). There was no statistically significant difference between the FEA-3@Mo group (69±5%) and the finasteride group. p >0.05).
[0044] Hair follicle transverse diameter: ImageJ measured the widest part of the hair follicle. The FEA-3@Mo group was 218±12μm, and the model group was 156±9μm, an increase of 40%. p <0.001).
[0045] Example 6: Molecular Mechanism Study Blind screening: Noni fruit extract FEA-3 fully binds to AR-LBD (PDB 1E3G), retaining binding energy <−9 kcalmol. -1 The trace components were screened and found to include ursolic acid, flavonoid X, and iridoid Y, indicating high efficacy at low content.
[0046] 1. Materials and Software Receptor crystal structure: PDB ID1E3G (human androgen receptor ligand binding domain, AR-LBD, resolution 1.65 Å, containing T877 mutation, complex with testosterone).
[0047] Reference ligand: DHT (PubChem CID10635, the original ligand in the crystal structure).
[0048] Test compound: Compound-2: Ursolic acid (UA), purity ≥98% (HPLC-UV220nm).
[0049] Compound-3: Flavonoid X (confirmed by NMR as 3,5,7,3′,4′-pentahydroxy-6-methylflavonoid, C16H12O7), prepared in-house, 96.8% pure.
[0050] Compound-4: Iridoid Y (identified as 6-O-trans-p-coumaryl-8-epio-loganic acid, C27H32O13 by HRMS and 1D / 2DNMR), prepared in-house, 95.4% pure.
[0051] Software platform: Schrödinger Suite 2023-3 (Maestro 13.9), Amber Tools 22, GROMACS 2022.4, PLIP 2.3.0, LigPlot+ 2.2.
[0052] 2. Protein pretreatment Use the Protein Preparation Wizard to remove water, add hydrogen, and complete the loop (Prime, deletion of residues 664-671).
[0053] The disulfide bonds Cys576-Cys650 and Zn²⁺ coordination tetrahedra (Cys507, Cys510, Cys613, Cys616) in the crystal structure are retained.
[0054] Protonation state was predicted using PROPKA 3.4 at pH 7.4; His741 was identified as the ε-tautomer.
[0055] Energy minimization (OPLS4 force field, RMSD converges to 0.30 Å).
[0056] Ligand preparation The 3D structure was generated by LigPrep, and the most likely protonation state (pH 7.4) was calculated by Epik. 32 stereoisomers / tautomers were generated, and the lowest energy conformation was used for docking.
[0057] Partial flexibility: The glycosidic bond (φ / ψ) of iridoid Y and the C-28 carboxylate group of ursolic acid are set as rotatable bonds (≤6 rotatable bonds, in accordance with Lipinski's rule).
[0058] 4. Molecular docking Lattice generation: Centered on the C17 oxygen atom of the crystal DHT, the box has a side length of 20 Å (covering the entire LBD pocket).
[0059] Precision mode: GlideSP to XP two-step strategy; XP mode additionally penalizes the solvent-exposed conformation.
[0060] Scoring function: GlideScore(GS) + MM - GBSAΔGbind; retain poses ≤ 1.0 Å heavy-atomRMSD.
[0061] Result threshold: XPGS ≤ −8.0 kcal mol -1 And MM-GBSA ΔGbind ≤ −9.0 kcal mol -1 It is considered a strong bond.
[0062] Molecular docking revealed that compounds 2 (identified as ursolic acid), 3 (flavonoid X), and 4 (iridocarboxylic acid Y) can stably bind to the androgen receptor ligand-binding domain (LBD) with a binding energy <-9.0 kcal / mol. MD simulations indicated that the complex was stable with an RMSD <2.0 Å.
[0063] Example 7: Validation of the targeting ability of the nanocomposite (FEA-3@Mo) to the androgen receptor Flow cytometry showed that the uptake rate of the nanocomplex (FEA-3@Mo) in dermal papilla cells with high androgen receptor expression was 3.2 times that of ordinary molybdenum nanoparticles. The dermal papilla cells with high androgen receptor expression were first incubated with the androgen receptor antagonist 10 μM enzalutamide at 37°C for 30 min, washed once with PBS, and then FITC-nanocomplex (FEA-3@Mo) (50 μg·m³) was added. -1 The samples were incubated at 37 °C for 1 h, washed three times with PBS, digested with trypsin, and FITC fluorescence was measured by flow cytometry. The fluorescence intensity of the antagonist group decreased by 70% compared with the NMNC single-addition group, which proves that the targeting is dependent on the androgen receptor.
[0064] Example 8: Comprehensive evaluation of pharmacodynamics and pharmacokinetics 1. Laboratory Animals and Ethics New Zealand white rabbits, male, 2.2 ± 0.2 kg, SPF grade, n = 18 (6 rabbits with drug-induced activity and 12 rabbits with irritant activity).
[0065] The experiment was approved by the Guangdong Medical University IACUC.
[0066] 2. Targeting coefficient (HF / plasma AUC ratio) 2.1 Drug Administration and Sampling: A 6 cm × 6 cm area of the shaved back was coated with 0.2 g of 5% NMNC gel (containing 10 μg Mo). Samples were collected at 0, 0.5, 1, 2, 4, 8, 12, 24, 48, and 72 h: ◦ Plasma: 1 mL of blood was collected from the marginal ear vein, anticoagulated with lithium heparin, and incubated at 3500 g for 10 min at 4℃. ◦ Hair follicle tissue: The animal was euthanized by air embolism after 72 h. Full-thickness skin from the treated area was harvested using a 6 mm punch, and the dermis and subcutaneous fat were separated using ophthalmic forceps and flash-frozen in liquid nitrogen.
[0067] 2.2 Sample Processing and Determination: Digestion: Add 0.2 mL of plasma or approximately 20 mg of hair follicle to 0.5 mL of 65% HNO3, digest at 120℃ for 30 min using graphite, and then bring the volume to 2 mL with deionized water. Quantification: Molybdenum was determined by ICP-MS (Agilent 7800) with internal standard. 95 Mo, LOD 0.02 ng mL⁻¹, recovery rate 92-1106%. Calculation: ◦ AUC0- 72 h (plasma) and AUC0- 72 h (hair follicle) was analyzed using the trapezoidal method (PKSolver 2.0). ◦ Targeting coefficient = AUC hair follicle / AUC plasma.
[0068] 2.3 Results: Plasma AUC = 370 ng·h·L⁻¹; Hair follicle AUC = 4.22 ng·h·g -1 Targeting coefficient = 11.4 (>5 indicates high enrichment).
[0069] 3. Systemic exposure (Cmax & PMTA comparison): Plasma Cmax = 42 ng·L -1 (≈0.042 μg·L) -1 ). PMTA (Permitted Maximum Tolerable Daily Intake for Mo) = 70 μg·kg -1 ·d -1 The acceptable daily dose for a 70 kg adult is 4.9 mg. In this study, a single transdermal Mo exposure = 10 μg / 2.2 kg rabbit ≈ 4.5 μg·kg -1 It is only 0.06% of PMTA; the cumulative exposure over 30 days is < 0.2% PMTA, indicating high system safety.
[0070] The targeting coefficient (HF / plasma AUC ratio) was 11.4, indicating that the hair follicles were highly enriched. System exposure Cmax = 42 ng·L -1 It is far below the acceptable daily intake of Mo (PMTA 70µg·kg) -1 ·d -1 ); The skin irritation index was 0.18 for 90 consecutive days (<0.4, which is considered non-irritating).
[0071] The above embodiments are merely preferred embodiments of the present invention and should not be construed as limiting the scope of protection of the present invention. Any non-substantial changes and substitutions made by those skilled in the art based on the present invention shall fall within the scope of protection claimed by the present invention.
Claims
1. A traditional Chinese medicine composition that prevents hair loss and promotes hair growth by inhibiting DHT receptor expression, characterized in that, The invention includes a nanocomposite formed by coordination-driven self-assembly of noni fruit extract FEA-3 and molybdate. The noni fruit extract FEA-3 acts as a reducing agent and stabilizer for molybdate, inducing the formation of molybdate-based nanoparticles with a particle size of 50-1200 nm. The noni fruit extract FEA-3 is rich in polyphenols and can specifically bind to androgen receptors on the surface of hair follicle cells, achieving targeted delivery of molybdate. The molybdate-based nanoparticles inhibit androgen receptor nuclear translocation and DHT binding within cells, blocking downstream signaling pathways of androgen receptors, thereby inhibiting hair follicle miniaturization and promoting hair growth.
2. The traditional Chinese medicine composition for preventing hair loss and promoting hair growth by inhibiting DHT receptor expression as described in claim 1, characterized in that, The noni fruit extract FEA-3 was prepared by the following method: noni fruit raw material was extracted with ethanol, concentrated, and then extracted with petroleum ether, ethyl acetate and n-butanol in sequence. The ethyl acetate layer was collected and then purified by macroporous resin column chromatography and gel column chromatography to obtain the noni fruit extract FEA-3.
3. The traditional Chinese medicine composition for preventing hair loss and promoting hair growth by inhibiting DHT receptor expression as described in claim 1, characterized in that, The molybdenum-based nanoparticles are introduced in the form of molybdate, which is selected from sodium molybdate, ammonium molybdate, or potassium molybdate.
4. The traditional Chinese medicine composition for preventing hair loss and promoting hair growth by inhibiting DHT receptor expression as described in claim 1, characterized in that, The nanocomposite has a particle size of 50-1150 nm and a zeta potential of -20 mV to -30 mV.
5. The traditional Chinese medicine composition for preventing hair loss and promoting hair growth by inhibiting DHT receptor expression as described in claim 1, characterized in that, The nanocomposite can synergistically inhibit the expression of androgen receptors, wherein: the noni fruit extract FEA-3 promotes the degradation of androgen receptor proteins by binding to the androgen receptor ligand-binding domain; after the molybdenum-based nanoparticles are released into cells, they downregulate the transcriptional level of androgen receptor genes, interfere with androgen receptor transcriptional activity, upregulate SULT-related genes, and downregulate the expression of iNOS, COX2 and Il1f8.
6. The traditional Chinese medicine composition for preventing hair loss and promoting hair growth by inhibiting DHT receptor expression as described in claim 1, characterized in that, The composition comprises the aforementioned nanocomposite, an active extract of a traditional Chinese medicine for treating androgenetic alopecia, and a pharmaceutically acceptable carrier, and the traditional Chinese medicine composition is used for topical treatment of androgenetic alopecia.
7. The traditional Chinese medicine composition for preventing hair loss and promoting hair growth by inhibiting DHT receptor expression as described in claim 6, characterized in that, The dosage form of the traditional Chinese medicine composition is a gel or cream.
8. A method for preparing a traditional Chinese medicine composition for preventing hair loss and promoting hair growth by inhibiting DHT receptor expression, as described in any one of claims 1-7, characterized in that, Includes the following steps: S1. The preparation of the nanocomposite includes the following steps: S11. Dissolve the noni fruit extract FEA-3 in the aqueous phase; S12, Add an aqueous solution containing the molybdate; S13. Nanoparticles are formed by self-assembly under ultrasonic or stirring conditions; S14. The nanocomposite was obtained by dialysis, concentration and lyophilization; S2. Compound of active components of traditional Chinese medicine: Take 10-30 parts of Platycladus orientalis leaves, 5-15 parts of Polygonum multiflorum, 5-15 parts of Ligustrum lucidum, and 3-10 parts of fresh ginger. Extract twice by reflux with 50-80% ethanol. Combine the filtrates and recover the ethanol under reduced pressure until there is no alcohol taste. Obtain a fluid extract with a relative density of 1.05-1.15, which is denoted as component B. S3, Composition Molding S31. Matrix preparation: Disperse 0.5-1.5 parts of carbomer 940, 5-10 parts of glycerol, and 0.02-0.1 parts of EDTA-2Na in deionized water at 60-80℃, and neutralize to pH 6.0-7.0 to obtain a transparent gel matrix. S32. Homogenization: 1-5 parts of the nanocomposite, 5-15 parts of component B, 0.1-0.5 parts of menthol, and 0.3-0.8 parts of preservative are added sequentially to the transparent gel matrix, and the mixture is homogenized under vacuum at 6000-12000 rpm for 3-10 min to remove bubbles. S33. Filling: Cool to below 30°C and fill into aluminum-plastic tubes or PET bottles under aseptic conditions to obtain the traditional Chinese medicine composition that achieves hair loss prevention and hair growth by inhibiting DHT receptor expression.