Application of nigella glandulifera seed extract in preparation of medicine for treating alopecia and daily necessities for promoting hair growth
By regulating VEGF, AR, and TGF-β1 through the extract of *Nepeta dentatum* seeds, a topical formulation was prepared to solve the treatment problem of androgenetic alopecia and achieve a safe and effective effect in promoting hair growth.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- SHANDONG ACADEMY OF PHARMACEUTICAL SCIENCES
- Filing Date
- 2026-03-11
- Publication Date
- 2026-05-12
AI Technical Summary
There is a lack of effective treatments for androgenetic alopecia in the current technology, and existing drugs have adverse drug reactions and compliance problems, especially the side effects caused by the use of minoxidil and finasteride.
Using extracts of *Nepeta glandulosa* seeds, topical formulations such as emulsions and microemulsions are prepared by regulating vascular endothelial growth factor (VEGF), inhibiting androgen receptor (AR), and transforming growth factor (TGF-β1) to promote hair growth.
It effectively promotes hair growth, avoids the side effects of traditional drugs, and has significant effects on promoting VEGF secretion, downregulating TGF-β gene expression, and reducing AR gene expression, thus having a significant therapeutic effect on hair loss.
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Abstract
Description
Technical Field
[0001] This invention relates to the field of extract technology of *Nigella glandularis* seeds, and more particularly to the application of *Nigella glandularis* seed extract in the preparation of drugs for treating hair loss and daily necessities for promoting hair growth. Background Technology
[0002] Hair loss is a common cosmetic dermatological condition, and its etiology may be related to genetic factors, psychological factors, hormone levels, sebum secretion, endocrine dysfunction, and trace element deficiencies. Studies have shown that most hair loss patients experience significant feelings of inferiority and depression, severely impacting their quality of life. Clinically, hair loss is broadly classified into diffuse and focal hair loss based on its underlying causes. Androgenetic alopecia (AGA) is the most common type of diffuse hair loss and the leading cause of hair loss in men, affecting approximately 70% of men at some point in their lives. Elevated levels of free testosterone and dihydrotestosterone (DHT) have been observed in men with AGA. Highly active DHT binds to androgen receptors in hair follicles, causing follicle atrophy and degeneration, leading to follicle miniaturization and ultimately hair loss. Minoxidil and finasteride are FDA-approved first-line drugs for treating hair loss. However, literature reports (e.g., Cai Changbin et al., *Chinese Journal of Drug Abuse Prevention and Treatment*, 2022, Vol. 28, No. 10; Cai Yiting et al., *Journal of Practical Dermatology*, 2025, Vol. 18, No. 4) indicate that long-term use of minoxidil has certain adverse drug reactions, mainly mild dermatitis, infection, and edema. Furthermore, 5% minoxidil is more likely to cause local irritation than 2% minoxidil, leading to contact pigmentary dermatitis and hirsutism, resulting in decreased compliance and tolerance. Accidental ingestion can cause cardiovascular adverse reactions, and the occurrence of adverse events is positively correlated with dosage. The contraindications of finasteride tablets clearly state that this drug is not suitable for women and children. Especially for pregnant women and women who may become pregnant, taking finasteride may cause abnormal development of the external genitalia in male fetuses. Furthermore, some literature reports (e.g., Sun Ke, Journal of Chronic Diseases, Vol. 12, No. 10, 2010) indicate that men who take finasteride orally to treat androgenetic alopecia may also develop finasteride syndrome after discontinuing the medication, which manifests as sexual dysfunction and psychological disorders, including decreased attention, depression, and suicidal tendencies.
[0003] Nigella glandularis seeds are from the Ranunculaceae family. Nigella glanduliferaThe dried, mature seeds of *Freyn et Sint.* are a traditional Uyghur medicinal material. Native to the Mediterranean region, they are also distributed in Central Asia of the former Soviet Union, primarily cultivated (Editorial Committee of Flora of China, Chinese Academy of Sciences: *Flora of China*, Vol. 27, p. 112, Science Press, July 1979, 1st edition). In China, they are found in Mengla, Jinghong, and Xinjiang (Kunming Institute of Botany, Chinese Academy of Sciences & Xishuangbanna Tropical Botanical Garden, Chinese Academy of Sciences: *List of Higher Plants of Xishuangbanna*, p. 81, Yunnan Nationalities Press, June 1996, 2nd edition). The 2020 edition of the *Chinese Pharmacopoeia* describes them as trigonous-ovate, 2.5-3 mm long and about 1.5 mm wide. The surface is black and rough, narrow and pointed at the apex, slightly blunt at the base, with irregular protrusions. They are hard, with a grayish-white, oily cross-section. They have a slightly fragrant odor and a pungent taste.
[0004] According to traditional Chinese medicine texts, *Nigeria galbana* seeds are used to dispel cold and reduce swelling, promote hair growth and darken hair, warm tendons and strengthen muscles, warm the uterus and relieve pain, and eliminate jaundice. They are used to treat various chronic inflammatory swellings caused by mucus, paralysis, facial paralysis, postpartum uterine pain, and jaundice. They can be used in preparations such as honey paste, digestive paste, powders, and oils. The 2020 edition of the *Chinese Pharmacopoeia* lists its functions as tonifying the kidneys and brain, regulating menstruation, promoting lactation, and diuresis. It is used for tinnitus, forgetfulness, amenorrhea, insufficient lactation, urinary tract infections, and urinary stones. Modern research on *Nigeria galbana* seeds is relatively limited. Domestic research mainly focuses on chemical composition studies, such as the chemical composition of *Nigeria galbana* seeds (a traditional Uyghur medicine from Xinjiang), and the isolation and structural identification of flavonoids in *Nigeria galbana* var. *galbana*. Pharmacological studies, such as the progress in chemical composition and pharmacological research of *Nigeria galbana* seeds, and quality studies, such as the determination of hederaponin and oleanolic acid content in *Nigeria galbana* var. *galbana* seeds by HPLC, and the high-performance liquid chromatography fingerprinting of *Nigeria galbana* var. *galbana* seeds.
[0005] Human dermal papilla cells (HDPCs) are differentiated from dermal connective tissue cells and are located at the base of hair follicles, possessing specific structures and functions. Vascular endothelial growth factor (VEGF) interacts with dermal papilla cells and hair follicle stem cells, promoting angiogenesis around the hair follicle and increasing nutrient supply to the hair follicle, thereby promoting hair growth. Dihydrotestosterone (DHT), on the other hand, increases AR expression, inhibits the translocation of β-catenin from the cytoplasm to the nucleus, and reduces the activity of hair follicle stem cells, leading to hair loss. DHT can also induce the hair growth inhibitory factor—transforming growth factor-β1 (TGF-β1)—inducing follicular fibrosis, thereby inhibiting the transition of hair from the telogen phase to the anagen phase. It is one of the key factors in the pathogenesis of androgenetic alopecia. Currently, there are no reported research data on the treatment of androgenetic alopecia with extracts of *Neatura odorata* seed. Summary of the Invention
[0006] The purpose of this invention is to address the aforementioned shortcomings of existing technologies by proposing the application of *Nigella glandularis* seed extract in the preparation of medications for treating hair loss and daily necessities for promoting hair growth. This effectively solves the problem that currently, there is no use of *Nigella glandularis* seed extract for treating androgenetic alopecia, both domestically and internationally. *Nigella glandularis* seed extract can achieve its therapeutic effect on hair loss by promoting vascular endothelial growth factor (VEGF), inhibiting androgen receptor (AR), and transforming growth factor-β1 (TGF-β1).
[0007] The first objective of this invention is to provide an application of a *Nigra galbana* seed extract in the preparation of a drug for treating hair loss, wherein the mass ratio of total saponins to total flavonoids in the extract is 6-14:1; the extract contains 14-21 mg / g of total flavonoids and 125-197 mg / g of total saponins.
[0008] Furthermore, the mass ratio of total saponins to total flavonoids in the extract is 8-13:1.
[0009] Furthermore, the extract contained 187.57 mg / g of total saponins and 15.05 mg / g of total flavonoids.
[0010] Furthermore, the extract is obtained by the following steps: oil extraction, extraction, concentration, macroporous resin adsorption, elution, concentration, water precipitation, and drying of black cumin seeds.
[0011] Further, the above preparation method can be carried out according to the following steps: Black cumin seeds are pressed using an oil press to obtain black cumin seed oil and black cumin seed cake; the black cumin seed cake is weighed, crushed, and 10-12 times its weight of 70% ethanol solution is added for extraction twice, 2 hours each time. The extract is concentrated to a relative density of 1.05, adsorbed onto HPD100 macroporous resin, and eluted sequentially with 3 times the resin column volume of water, 3 times the volume of 30% ethanol, and 4 times the volume of 70% ethanol. The 30% ethanol eluent is concentrated under reduced pressure to a relative density of 1.20-1.25, and 5 times the volume of ethanol is added. Add water, stir and filter, concentrate and dry the filtrate to obtain total flavonoid extract of *Nigella glandularis* seeds; concentrate the 70% alcohol eluent under reduced pressure to a relative density of 1.15-1.20, add 5 times the volume of water, stir and filter, concentrate and dry the filtrate to obtain total saponin extract of *Nigella glandularis* seeds; mix the total flavonoid extract and total saponin extract in a certain proportion or combine the 30% and 70% alcohol eluents, concentrate under reduced pressure to a relative density of 1.15-1.20, add 5 times the volume of water, stir and filter, concentrate and dry the filtrate to obtain extract of *Nigella glandularis* seeds.
[0012] Furthermore, medications for treating androgenetic alopecia.
[0013] Furthermore, the medicine is a topical preparation.
[0014] Furthermore, the dosage form of the topical preparation is an emulsion, microemulsion, submicroemulsion, gel, powder, suspension, cream, gel, or spray.
[0015] Furthermore, the daily necessities mentioned are hair sprays or shampoos.
[0016] This invention utilizes a method of extraction followed by macroporous resin adsorption purification after oil extraction from black cumin seeds to enrich an extract containing saponins and flavonoids. This method effectively enriches the saponins and flavonoids in black cumin seeds without affecting the extraction of black cumin seed oil. This invention replaces traditional organic solvent extraction methods, avoiding organic solvent contamination, and offers advantages such as low production cost, high content, ease of operation, and suitability for industrial production.
[0017] The mechanism of action of the extract of *Neema galbana* seeds in this invention is to promote vascular endothelial growth factor (VEGF), inhibit androgen receptor (AR) and transforming growth factor (TGF-β1) to achieve the effect of treating hair loss, which is the first of its kind in current domestic and international research.
[0018] To determine the optimal mass ratio of total saponins to total flavonoids in this invention, extracts with mass ratios of 5:1, 8:1, 10:1, 13:1, and 15:1 were prepared. Cell viability was tested using conventional cell viability assays (2 mg / mL concentration). Through screening, it was found that extracts with a mass ratio of total saponins to total flavonoids within the range of 8:1 to 13:1 showed the most significant improvement in DHT-induced cell viability inhibition (cell viability >120%), and exhibited superior VEGF secretion promotion and AR gene downregulation compared to other ratios. These extracts demonstrated a synergistic enhancing effect in promoting dermal papilla cell viability, stimulating VEGF secretion, and regulating the expression of β-Catenin, AR, and TGF-β genes, with significantly better effects than extracts outside this ratio range. Detailed Implementation
[0019] The following are specific embodiments of the present invention, which further describe the technical solution of the present invention, but the present invention is not limited to these embodiments.
[0020] Example 1 The extract of *Nigella glandularis* seeds was obtained as follows: 1 kg of *Nigella glandularis* seeds were weighed and pressed using an oil press to obtain *Nigella glandularis* seed oil and *Nigella glandularis* seed cake; 300 g of *Nigella glandularis* seed cake was weighed, crushed, and extracted twice with 12 times its weight of 70% ethanol solution for 2 hours each time. The extract was concentrated to a relative density of 1.05, and adsorbed onto HPD100 macroporous resin. Elution was performed sequentially with 3 times the resin column volume of water, 3 times the volume of 30% ethanol, and 4 times the volume of 70% ethanol. The 30% and 70% ethanol eluates were combined and concentrated under reduced pressure to a relative density of 1.15-1.20. 5 times the volume of water was added, and the mixture was stirred and filtered. The filtrate was concentrated and dried to obtain the extract of *Nigella glandularis* seeds.
[0021] Example 2 The extract of *Nigella glandularis* seeds was obtained as follows: 1 kg of *Nigella glandularis* seeds were weighed and pressed using an oil press to obtain *Nigella glandularis* seed oil and *Nigella glandularis* seed cake; 300 g of *Nigella glandularis* seed cake was weighed, crushed, and extracted twice with 12 times its weight of 70% ethanol solution, 2 hours each time. The extract was concentrated to a relative density of 1.05, adsorbed onto HPD100 macroporous resin, and eluted sequentially with 3 times the resin column volume of water, 3 times the volume of 30% ethanol, and 4 times the volume of 70% ethanol. The alcohol eluent was concentrated under reduced pressure to a relative density of 1.20-1.25, 5 times its volume of water was added, the mixture was stirred and filtered, and the filtrate was concentrated and dried to obtain the total flavonoid extract of *Nigella glandularis* seeds; the 70% alcohol eluent was concentrated under reduced pressure to a relative density of 1.15-1.20, 5 times its volume of water was added, the mixture was stirred and filtered, and the filtrate was concentrated and dried to obtain the total saponin extract of *Nigella glandularis* seeds; the total flavonoid extract and the total saponin extract were mixed in a certain proportion to obtain the *Nigella glandularis* seed extract.
[0022] Content determination methods and results: (1) Determination of total flavonoid content: Kaempferol, the main characteristic component of the hydrolyzed black cumin seed extract, was used as a reference. The absorbance of the reference at different concentrations was determined by ultraviolet spectrophotometry at 265 nm. A standard curve was plotted based on the relationship between absorbance and concentration. The black cumin seed extract was dissolved in 50% ethanol, and an appropriate amount of hydrochloric acid was added. The mixture was refluxed, cooled, and then extracted multiple times with chloroform. The chloroform layer was evaporated to dryness, and the residue was diluted with 50% ethanol to a certain volume. The absorbance was then determined.
[0023] Based on the obtained absorbance, the content of the reference standard can be obtained from the standard curve, and the total flavonoid content in the black cumin seed extract obtained in Example 1 is calculated to be 15.05 mg / g.
[0024] Based on the obtained absorbance, the content of the reference standard can be obtained from the standard curve, and the total flavonoid content in the total flavonoid extract obtained in Example 2 is calculated to be 33.35 mg / g.
[0025] (2) Determination of total saponin content: The main characteristic component of the hydrolyzed black cumin seed extract, ivy saponin, was used as a reference. It was dissolved in vanillin-glacial acetic acid solution, color developed with perchloric acid, and then diluted to volume with glacial acetic acid. The absorbance of the reference at different concentrations was determined by ultraviolet spectrophotometry at 545 nm. A standard curve was plotted based on the relationship between absorbance and concentration. The black cumin seed extract was dissolved in 50% ethanol, and an appropriate amount of hydrochloric acid was added. After reflux and cooling, it was extracted multiple times with chloroform. The chloroform layer was evaporated to dryness, and the residue was diluted to volume with 50% ethanol. An appropriate amount was evaporated to dryness, dissolved in vanillin-glacial acetic acid solution, color developed with perchloric acid, and then diluted to volume with glacial acetic acid. The absorbance was then determined.
[0026] Based on the obtained absorbance, the content of the reference standard can be obtained from the standard curve, and the total saponin content in the black cumin seed extract obtained in Example 1 is calculated to be 187.57 mg / g.
[0027] Based on the obtained absorbance, the content of the reference standard can be obtained from the standard curve, and the total saponin content in the total saponin extract obtained in Example 2 is calculated to be 338.78 mg / g.
[0028] The above methods for determining total flavonoids and total saponins have all undergone systematic adaptability and methodological investigation, and have excellent accuracy, stability and reproducibility, and can accurately determine the content of each component in black cumin seed extract.
[0029] efficacy trial for treating hair loss A male pattern baldness model was established by inducing human dermal papilla cells with dihydrotestosterone to study the efficacy of extracts from *Neema galbana* seeds in treating hair loss. The specific experiment is as follows.
[0030] 1. Experimental materials and reagents Human dermal papillary cells (HDPCs), DMEM medium, fetal bovine serum, antibiotics, trypsin, PBS buffer, CCK-8 kit, RNA extraction solution, dihydrotestosterone, minoxidil, Human VEGF ELISA kit, cell culture plates.
[0031] 2. Main Equipment Clean bench, CO2 incubator, pipettes, multi-functional microplate reader, real-time quantitative PCR instrument.
[0032] 3. Sample preparation The samples were diluted to the initial concentration using culture medium, then filtered through a 0.22-micron filter membrane for sterilization, and finally diluted to the test concentration using culture medium.
[0033] 4. Cell viability test - sample Cells were cultured normally under conditions of 5% CO2 and 37°C. When the cells reached 60-70% confluence, they were digested with trypsin and seeded into 96-well plates. After 24 hours of cell adhesion, different concentrations of the test samples were added, and a blank control group was set up. After 24 hours of incubation, cell viability was measured (CCK-8 method). Each experimental group had 3 parallel wells, and the relative cell viability was calculated according to the following formula: Relative cell activity % = Absorbance of sample group / Absorbance of blank control group × 100%.
[0034] By conducting cell viability tests, samples with safe concentrations (cell viability greater than 80%) that do not affect cell viability are selected for the next stage of efficacy testing.
[0035] 5. Cell viability assay - DHT Cells were cultured normally under conditions of 5% CO2 and 37°C. When the cells reached 60-70% confluence, they were digested with trypsin and seeded into 96-well plates. After 24 hours of cell adhesion, different concentrations of dihydrotestosterone (DHT) were added, with a blank control group included. After 24 hours of incubation, cell viability was measured (CCK-8 assay). Each experimental group had three parallel wells, and relative cell viability was calculated using the following formula: Relative cell activity % = Absorbance of sample group / Absorbance of blank control group × 100%.
[0036] 6. Cell viability assay - Effect of samples on DHT-induced cell viability Cells were cultured normally under conditions of 5% CO2 and 37°C. When the cells reached 60-70% confluence, they were digested with trypsin and seeded into 96-well plates. After 24 hours of cell adhesion, 50 μg / ml of dihydrotestosterone and different concentrations of the test samples were added, with a blank control group included. After 24 hours of incubation, cell viability was measured (CCK-8 assay). Each experimental group had three parallel wells, and relative cell viability was calculated using the following formula: Relative cell activity % = Absorbance of sample group / Absorbance of blank control group × 100%.
[0037] 7. Effects of the sample on DHT-induced cell-related genes and proteins Cells were cultured normally under conditions of 5% CO2 and 37°C. When the cells reached 60-70% confluence, they were digested with trypsin and seeded into 96-well plates. After 24 hours of cell adhesion, the culture medium was discarded, and the medium was divided into solvent control group, model control group (DHT 50 μg / L), sample group (containing DHT 50 μg / L and samples of different concentrations), and positive control group (containing DHT 50 μg / L and minoxidil 10 μg / mL), and incubated for 24 hours.
[0038] VEGF: Collect cell supernatant after 24 hours and freeze. Measure the VEGF concentration in the culture supernatant according to the ELISA kit instructions.
[0039] TGF-β, β-Catenin, and AR gene detection: After 24 hours of culture, the supernatant was discarded, and the cells were washed twice with pre-cooled phosphate-buffered saline (PBS). Cell lysis buffer was then prepared using RNA extraction reagent. Total RNA was extracted from all lysis buffer samples, reverse transcribed into cDNA, and gene expression levels were detected using real-time quantitative PCR.
[0040] The primer sequences for the PCR test are shown in the table below.
[0041] PCR test primer sequences
[0042] 8. Statistical Methods The data were statistically processed using specialized software, and analysis of variance was used to compare statistical differences, with pairwise comparisons performed. P A value < 0.05 is considered statistically significant. * indicates... P <0.05, ** represents P <0.01.
[0043] Genetic data statistics - △△CT method: A = CT(target gene, sample to be tested) - CT(internal standard gene, sample to be tested) B = CT (target gene, control sample) - CT (internal standard gene, control sample) K=AB Expression multiple = 2 *-（K） 9. Test Results To determine the optimal mass ratio of total saponins to total flavonoids in this invention, the total flavonoid extract and total saponin extract obtained in Example 2 were mixed in a certain proportion. The mass ratio of total saponins to total flavonoids and the content of each component in the black cumin seed extract are shown in the table below.
[0044] Extract components
[0045] 9.1 Effect of Samples on Cell Viability Cell viability was assessed using samples with mass ratios of 5:1 and 15:1 to examine the dosage of the administered samples. At a concentration of 5 mg / ml, cell viability in both groups was significantly lower than 80%, indicating an impact on cell activity. At concentrations of 2 mg / ml, 1 mg / ml, 0.5 mg / ml, and 0.1 mg / ml, cell viability was higher than 80%, showing no cytotoxicity.
[0046]
[0047] 9.2 Effects of dihydrotestosterone on cell viability Compared to the blank control, the cell viability of dihydrotestosterone (DHT) at concentrations of 100 μg / mL and 50 μg / mL was significantly lower than 80%, indicating an impact on cell viability. A cell model was established using DHT at a concentration of 50 μg / mL to prevent cell detachment. The results are shown in the table below: Analysis of cell activity and differential significance of dihydrotestosterone-treated samples
[0048] Note: The presence of precipitate at a concentration of 100 μg / mL in cells indicates that the solubility has been exceeded and the test is not suitable.
[0049] 9.3 Effect of dihydrotestosterone on cell viability Both the minoxidil group and the sample group showed higher cell activity at the tested concentrations than the model group (dihydrotestosterone), and no cytotoxicity. At a 10:1 ratio relative to the model group (dihydrotestosterone), the minoxidil group and the sample group (**) showed significantly higher cell activity. P <0.01), and when the sample ratios are 8:1 and 13:1 (* P <0.05), significantly improved cell activity, indicating that the sample (8:1 to 13:1 ratio) and minoxidil can improve the inhibitory effect of dihydrotestosterone on cell activity.
[0050] Analysis of cell activity and differential significance of dihydrotestosterone-treated samples
[0051] 9.4 Effects of the sample on dihydrotestosterone-induced cell-related genes and proteins VEGF content: Compared to the control group, the VEGF content in the model group (dihydrotestosterone) was significantly reduced (**). P <0.01); compared to the model group (dihydrotestosterone), the VEGF content in the minoxidil group was 23.88±2.36 ng / ml (** P <0.01), which can significantly promote VEGF secretion by cells; the VEGF content in the sample group at 10:1 was 24.87±1.89 ng / ml (** P <0.01), the VEGF content at 13:1 was 24.59±0.67 ng / ml (** P <0.01), at an 8:1 ratio, the VEGF content was 19.99±0.69 ng / ml (* P <0.05), can significantly promote the secretion of VEGF in cells.
[0052] VEGF content and significance analysis
[0053] TGF-β gene detection: Compared to the model group (dihydrotestosterone), the TGF-β expression level in the minoxidil group was significantly reduced, with a gene expression rate of 0.84 ± 0.07 (**). P <0.01), indicating a downregulation effect of TGF-β; the expression level of the sample group was significantly reduced at ratios of 8:1 and 10:1, with a gene expression rate of 0.83±0.08 (**). P <0.01) and 0.82±0.05 (** P <0.01), has a TGF-β downregulatory effect.
[0054] TGF-β gene and differential significance analysis
[0055] β-Catenin gene detection: Compared to the model group (dihydrotestosterone), the minoxidil group showed no significant difference in β-Catenin expression levels, with no upregulation effect; however, the β-Catenin expression levels in the sample groups were significantly increased at ratios ranging from 5:1 to 13:1 (**). P <0.01 or * P <0.05).
[0056] β-Catenin gene and differential significance analysis
[0057] AR gene testing: Compared with the model group (dihydrotestosterone), the AR expression level in the minoxidil group was not significantly different and had no downregulation effect; the AR expression level in the sample groups was significantly reduced at the ratios of 8:1 / 10:1 / 13:1, with gene expression rates of 0.75±0.09 (**P<0.01), 0.66±0.10 (**P<0.01), and 0.78±0.12 (*P<0.05), respectively, showing a downregulation effect.
[0058] AR gene and differential significance analysis
[0059] 9.5 Experimental Conclusion This experiment established a male pattern baldness model using dihydrotestosterone-induced human dermal papilla cells to study the in vitro cellular activity of *Nitraria tangutorum* seed extract. The mechanism of action was investigated by measuring VEGF secretion, TGF-β, β-Catenin, and AR gene expression levels. Results showed that, compared to minoxidil, the extracts of this invention in the ratio range of 8:1 to 13:1 also significantly promoted VEGF secretion and downregulated TGF-β gene expression. Furthermore, it possessed the ability to increase β-Catenin gene expression and decrease AR gene expression, which minoxidil lacked. This indicates that the *Nitraria tangutorum* seed extract of this invention promotes hair follicle growth and development and has therapeutic effects on hair loss.
[0060] For any points not covered above, existing technologies shall apply.
[0061] Although specific embodiments of the present invention have been described in detail by way of examples, those skilled in the art should understand that the above examples are for illustrative purposes only and are not intended to limit the scope of the invention. Those skilled in the art can make various modifications or additions to the described specific embodiments or use similar methods to replace them, without departing from the direction of the invention or exceeding the scope defined by the appended claims. Those skilled in the art should understand that any modifications, equivalent substitutions, improvements, etc., made to the above embodiments based on the technical essence of the present invention should be included within the protection scope of the present invention.
Claims
1. The application of a *Nepeta glandulosa* seed extract in the preparation of drugs for treating hair loss and daily necessities for promoting hair growth, characterized in that, The extract has a total saponin to total flavonoid mass ratio of 6-14:1; the extract contains 14-21 mg / g of total flavonoids and 125-197 mg / g of total saponins.
2. The application as described in claim 1, characterized in that, The extract has a total saponin to total flavonoid mass ratio of 8-13:1, and the extract contains 14-19 mg / g of total flavonoids and 149-191 mg / g of total saponins.
3. The application as described in claim 1, characterized in that, The extract contained 187.57 mg / g of total saponins and 15.05 mg / g of total flavonoids.
4. The application as described in claim 1, characterized in that, The extract is prepared by the following steps: pressing, extracting, concentrating, adsorbing with macroporous resin, eluting, concentrating, water precipitation, and drying of black cumin seeds to obtain the black cumin seed extract.
5. The application as described in claim 1, characterized in that, The above preparation method can be carried out according to the following steps: Black cumin seeds are pressed using an oil press to obtain black cumin seed oil and black cumin seed cake; the black cumin seed cake is weighed, crushed, and extracted twice with 10-12 times its weight of 70% ethanol solution for 2 hours each time. The extract is concentrated to a relative density of 1.05, adsorbed onto HPD100 macroporous resin, and eluted sequentially with 3 times the resin column volume of water, 3 times the 30% ethanol, and 4 times the 70% ethanol. The 30% ethanol eluent is concentrated under reduced pressure to a relative density of 1.20-1.25, and 5 times its volume of... Water, stir and filter, concentrate and dry the filtrate to obtain total flavonoid extract of *Nigella glandularis* seeds; concentrate the 70% alcohol eluent under reduced pressure to a relative density of 1.15-1.20, add 5 times the volume of water, stir and filter, concentrate and dry the filtrate to obtain total saponin extract of *Nigella glandularis* seeds; mix the total flavonoid extract and total saponin extract in a certain proportion or combine the 30% and 70% alcohol eluents and concentrate under reduced pressure to a relative density of 1.15-1.20, add 5 times the volume of water, stir and filter, concentrate and dry the filtrate to obtain extract of *Nigella glandularis* seeds.
6. The application as described in claim 1, characterized in that, Medications for treating androgenetic alopecia.
7. The application as described in claim 1, characterized in that, The drug is a topical preparation, and the dosage form of the topical preparation is an emulsion, microemulsion, submicroemulsion, gel, powder, suspension, cream, gel, or spray.
8. The application as described in claim 1, characterized in that, The daily necessities mentioned are hair sprays or shampoos.