New Uses of Atractylodes macrocephala essential oil for hair growth

Abstract

This invention provides a novel use of Atractylodes macrocephala essential oil for the prevention and treatment of hair loss and alopecia areata. The characteristic components of the Atractylodes macrocephala essential oil include: atractylenolide I, atractylenolide II, atractylenolide III, atractylone, β-elemene, hesperidin, and isohesperidin. The invention also provides a method for preparing essential oil compositions, pharmaceutical formulations, and uses for the prevention and treatment of hair loss and alopecia areata. This method reduces bitterness and the first-pass effect in the liver, resulting in more stable blood drug concentrations and improved patient compliance. Compared to existing methods, it has advantages such as better efficacy and no toxic side effects. Developing health products and pharmaceuticals for the prevention and treatment of hair loss and alopecia areata based on this method has great potential.

Description

Technical Field

[0001] This invention relates to a therapeutic technique for promoting hair growth, specifically a novel application of Atractylodes macrocephala essential oil for hair growth. It belongs to the field of traditional Chinese medicine technology. Background Technology

[0002] Hair loss is a common skin condition characterized by thinning hair. Modern medicine attributes it to impaired immune function, skin diseases, and environmental factors, while Traditional Chinese Medicine (TCM) attributes it to liver and kidney deficiency, insufficient essence and blood, damp-heat accumulation, and blood stasis obstructing the pores. According to the latest scalp health survey report, "China Scalp Health White Paper," 250 million people in China suffer from hair loss, with prevalence rates of 70% for men and 40% for women. In terms of China's total population, this means that on average, one in six Chinese people experiences hair loss. Among them, 84% experience hair loss before the age of 30, 20 years earlier than the previous generation, indicating that hair loss is increasingly affecting younger people. Another resident health survey report shows that hair loss ranks seventh among health concerns for Chinese people. Whether from a health or appearance perspective, hair loss has become a top priority for contemporary young people. The most common types of hair loss in clinical practice today include: stress-related hair loss, physical hair loss, chemical hair loss, symptomatic hair loss, nutritional supplement-induced hair loss, infectious hair loss, endocrine-related hair loss, alopecia areata, and gray hair.

[0003] In recent years, receding hairlines and sudden graying of the hairline have plagued many modern middle-aged people. Hair loss is related to many factors, including genetics, autoimmune disorders, and psychological stress. With changing lifestyles and increased work pressure, hair loss is affecting a growing number of modern people. It severely impacts a person's image, causing a loss of self-confidence and even hindering interpersonal relationships and employment. Patients hope to resolve their hair loss problems through hair regrowth products or even medications. While there are many hair care and regrowth products on the market, their development is not yet fully mature. Therefore, developing more effective and popular hair regrowth products would undoubtedly bring significant value.

[0004] Currently, the main treatment for hair loss is medication, with topical minoxidil being widely used clinically for alopecia areata. Prostaglandins and platelet-rich plasma (PRP) also promote hair growth. Another study validated the higher efficacy and feasibility of using 308nm excimer laser combined with minoxidil for treating alopecia areata. In addition, the anti-inflammatory effects of JAK kinase inhibitors and statins can target inflammation, the main pathology of alopecia areata, and have shown good efficacy. The specific targeted therapy of biologics such as abatacept, INF-γ antibodies, and apremilast can reduce the levels of major inflammatory factors like INF-γ, proving effective, but the efficacy remains to be verified. Tamoxetine and α-MSH analogs can restore the immune clearance of hair follicles, thereby treating alopecia areata. Foreign studies have confirmed that CRHR antagonists Antalarmin and Astressin can improve anxiety symptoms by reversing excessive activity of the HPA axis, and also have therapeutic effects suitable for inflammatory skin diseases such as alopecia areata. Minoxidil is mainly used for androgenetic alopecia, finasteride is mainly for androgenetic alopecia, but it produces certain side effects. Spironolactone's main function is anti-androgen. All of these treatments have varying degrees of side effects, poor efficacy, slow onset of action, significant toxicity and instability, easy relapse, and certain limitations, seriously affecting their current clinical use. Using traditional Chinese medicine formulas to solve modern problems can help develop better products with fewer side effects.

[0005] Atractylodes macrocephala Koidz., the dried rhizome of the Asteraceae plant Atractylodes macrocephala, has the effects of invigorating the spleen and replenishing qi, drying dampness and promoting diuresis, stopping sweating, and calming the fetus. It is mainly used for spleen deficiency with poor appetite, abdominal distension and diarrhea, phlegm retention with dizziness and palpitations, edema, spontaneous sweating, and threatened abortion. Modern pharmacological studies have shown that Atractylodes macrocephala has multiple effects such as diuresis, anti-tumor, anti-inflammatory, anti-diabetic, and anti-aging. Atractylodes macrocephala essential oil is extracted from Atractylodes macrocephala. Current reports on the application of Atractylodes macrocephala essential oil are limited to its good effects in antibacterial and anti-tumor aspects, and there are no reports on its hair growth and nourishing effects. Summary of the Invention

[0006] In view of this, the purpose of this invention is to propose a new use for Atractylodes macrocephala essential oil, and to provide an application of Atractylodes macrocephala essential oil in the preparation of health products, pharmaceuticals, medical devices or daily chemical products for the prevention or treatment of hair loss and alopecia areata.

[0007] To achieve the above objectives, the first aspect of the present invention provides an essential oil for preventing or treating hair loss, characterized in that the essential oil is Atractylodes macrocephala essential oil.

[0008] Herbal essential oils are volatile, aromatic liquids extracted from the flowers, leaves, stems, roots, or fruits of medicinal herbs. They are immiscible with water and possess a fragrant aroma. Inherited from traditional Chinese medicine theory, they play an important therapeutic role in many Chinese medicines and are indispensable active ingredients. Essential oils are mainly composed of chemical molecules such as terpenes, aldehydes, alcohols, esters, ketones, and phenols. Most have antibacterial, antiviral, antimicrobial, and anti-inflammatory effects. At the same time, essential oils are also powerful antioxidants, effectively resisting oxidative stress, regulating human physiological functions, reducing the attack of free radicals on human tissues, enhancing immunity, coordinating endocrine function, aiding in the normal functioning of digestive and circulatory systems, and helping the body restore and maintain a healthy state.

[0009] The Atractylodes macrocephala essential oil preparation of the present invention exerts its drug tropism and functional effects through inhalation and transdermal absorption, improves symptoms such as hair loss and alopecia areata, avoids the bitterness of oral drugs and the first-pass effect of the liver, has a more stable blood drug concentration, reduces the pain of injecting drugs, and improves patient compliance.

[0010] The Atractylodes macrocephala mentioned refers to the tuberous root of Atractylodes macrocephala Koidz., a plant of the Asteraceae family, and its cultivated varieties.

[0011] The Atractylodes macrocephala essential oil preparation may also include pharmaceutically acceptable excipients, such as transdermal absorbents, fillers, binders, surfactants, wetting agents, emulsifiers, suspending agents, preservatives, or combinations thereof; the transdermal absorbent may be a traditional Chinese medicine essential oil or a traditional Chinese medicine including peppermint, borneol, etc.; the filler may be selected from dextrin, starch, sorbitol, lactose, mannitol, xylitol, microcrystalline cellulose, etc.; the binder may be selected from cellulose derivatives, vinyl acetate resin, polyvinyl alcohol, gelatin, alginate, etc.; the surfactant may be selected from sodium dodecylbenzenesulfonate, stearic acid, polyoxyethylene-polyoxypropylene copolymer, fatty acid sorbitan, or polysorbate (Tween), etc.

[0012] Alternatively, Atractylodes macrocephala essential oil is a volatile oil that is insoluble in water and has an aromatic odor, obtained by extracting Atractylodes macrocephala tuberous roots through supercritical CO2 extraction, subcritical fluid extraction, or steam distillation.

[0013] Optionally, the characteristic components of the Atractylodes macrocephala essential oil include seven components: atractylodes macrocephala lactone I, atractylodes macrocephala lactone II, atractylodes macrocephala lactone III, atractylone, β-elemene, hesperidin and isohesperidin.

[0014] Optionally, the Atractylodes macrocephala essential oil preparation includes a dosage form made with pharmaceutically acceptable excipients for convenient administration.

[0015] For the same purpose, a second aspect of the present invention provides a method for preparing the essential oil formulation for preventing or treating hair loss as described in the first aspect of the present invention, comprising the following steps: compounding Atractylodes macrocephala essential oil with an organic solvent and a transdermal absorption agent to obtain the essential oil complex.

[0016] Optionally, the organic solvent may include one or more of alcohol, propylene glycol, glycerin, base oil, n-butane, and mineral oil; the base oil may be one or more of jojoba oil, sweet almond oil, grapeseed oil, rosehip oil, and olive oil; the transdermal absorbent may be one or more of traditional Chinese medicine essential oils and extracts; the traditional Chinese medicine essential oils may be dried ginger, fennel, mugwort, cinnamon, and peppermint essential oils; and the traditional Chinese medicine extracts may be borneol, camphor, etc., without specific limitations.

[0017] Optionally, the proportion of alcohol in the essential oil composition shall not exceed 50% to ensure safety. Optionally, the alcohol may be dealdehyde alcohol, which, by utilizing its odorless property, can effectively avoid the influence of the alcohol's own smell on the aroma of the essential oil composition.

[0018] In this invention, when preparing the essential oil composition, the raw material of Atractylodes macrocephala is extracted with Atractylodes macrocephala essential oil by supercritical CO2 extraction, subcritical fluid extraction or steam distillation extraction, and then set aside for later use.

[0019] In some possible implementations, the process of extraction using supercritical extraction technology is as follows: Atractylodes macrocephala root is crushed and added to a material cylinder, which is then placed into an extraction tank, sealed tightly, and subjected to supercritical CO2 extraction to obtain Atractylodes macrocephala essential oil; wherein the process parameters are: particle size of 40-60 mesh, extraction pressure of 15-30 MPa, extraction temperature of 30-50℃, extraction time of 2-4.5 hours, CO2 flow rate of 15-40 L / h, and the extract is obtained by washing and separating with water after releasing carbon dioxide.

[0020] In some possible embodiments, the process for extracting Atractylodes macrocephala essential oil using subcritical extraction is as follows: Atractylodes macrocephala root is crushed and placed in an extraction tank. After vacuuming, solvent is added to the extraction tank, with the amount of solvent being 5-10 times the amount of raw material. The temperature is 35-55℃, the pressure is 0.3-0.8MPa, and the extraction time is 1-8 hours. The solvent is butane. After extraction, the discharge valve at the bottom of the extraction tank is opened, and the extract is transferred to an evaporator to obtain the subcritical primary extract of Atractylodes macrocephala. The solvent is compressed and recovered to a solvent tank by a circulating compressor for reuse. This process is repeated multiple times, and the subcritical primary extracts are combined. The obtained primary extract is then subjected to rotary evaporation to recover the solvent, heated with ethanol for reconstitution, frozen, filtered, and the filtrate is subjected to rotary evaporation to obtain Atractylodes macrocephala essential oil.

[0021] In some possible implementations, the process for extracting Atractylodes macrocephala essential oil by steam distillation is as follows: particle size: 60-120 mesh, according to Appendix XD of the 2010 edition of the Chinese Pharmacopoeia, volatile oil determination method (method A), first soak for 2.5-6 hours, then distill for 5-9 hours, and obtain the oil after oil-water separation.

[0022] It should be noted that supercritical CO2 extraction, subcritical fluid extraction, and steam distillation are merely examples. Those skilled in the art can select appropriate volatile oil extraction methods according to actual conditions, including but not limited to the methods listed above. For different pharmaceutical raw materials, those skilled in the art can make appropriate adjustments to the above conditions, without any specific limitations.

[0023] For the same purpose, a third aspect of the present invention provides the application of preparations made from essential oils for the prevention or treatment of hair loss as described in the first aspect as health products, pharmaceuticals, medical devices or daily chemical products for the prevention or treatment of hair loss.

[0024] Optionally, the dosage form of the formulation includes a nasal administration formulation or a transdermal administration formulation. Nasal and transdermal administration can directly enter the bloodstream, have a faster onset of action, reduce the burden on the patient's gastrointestinal tract and liver, and are suitable for patients with impaired gastrointestinal function, showing broad application prospects.

[0025] Optionally, the dosage form of the preparation includes fumigants, inhalers, sprays, aerosols, ointments, patches, liniments, tinctures, lotions, oils, films, drops, or suppositories.

[0026] Experiments have shown that this essential oil preparation can enter the bloodstream and digestive system through the nose and skin, thereby regulating the body's endocrine and digestive systems. Animal experiments have shown that after intervention with Atractylodes macrocephala essential oil, the physiological indicators of the model group mice, such as hair growth tendency, hair growth quality, and hair texture, have significantly improved. Human experiments have also confirmed that Atractylodes macrocephala essential oil products can significantly improve symptoms of hair loss and alopecia areata. Based on this, there is great potential for developing products with hair care and nourishing effects.

[0027] As can be seen from the above, the Atractylodes macrocephala essential oil provided by the present invention is used for the prevention or treatment of hair loss, its preparation method, formulation and uses. The essential oil can prevent or treat hair loss and alopecia areata. Moreover, the essential oil can achieve the effect of preventing or treating hair loss and alopecia areata by both nasal and transdermal administration, reducing the bitterness of oral drugs and the first-pass effect of the liver, making the blood drug concentration more stable, reducing the pain of patients injecting drugs, and improving patients' compliance with medication. Detailed Implementation

[0028] To make the objectives, technical solutions, and advantages of this disclosure clearer, the following detailed description is provided in conjunction with specific embodiments.

[0029] It should be noted that, unless otherwise defined, the technical or scientific terms used in the embodiments of the present invention should have the ordinary meaning as understood by one of ordinary skill in the art to which this disclosure pertains.

[0030] Example

[0031] I. Extraction of Atractylodes macrocephala essential oil

[0032] The dried Atractylodes macrocephala raw material was pulverized using a pulverizer, passed through a 40-mesh sieve, weighed, and loaded into a supercritical extraction vessel for extraction. The vessel was then sealed tightly. The extraction pressure was set to 30 MPa, the extraction temperature to 40 °C, the extraction time to 2.0 h, and the CO2 flow rate to 15 L / h. Atractylodes macrocephala essential oil was obtained and set aside for later use.

[0033] II. Preparation of Atractylodes macrocephala essential oil preparations

[0034] The examples are Atractylodes macrocephala essential oil and excipients, with Examples 1-3 being Atractylodes macrocephala and olive oil (base oil), and Example 4 being a mixture of Atractylodes macrocephala essential oil and a traditional Chinese medicine extract (transdermal absorption agent). The traditional Chinese medicine extract is borneol. The formulation is shown in Table 1.

[0035] Table 1

[0036]

[0037] Example: Preparation of essential oils:

[0038] 1. Weigh 1g of Atractylodes macrocephala essential oil and dissolve it in 10g of olive oil. Then mix the resulting liquids completely to obtain the essential oil of Example 1.

[0039] 2. Weigh 0.1g of Atractylodes macrocephala essential oil and dissolve it in 10g of olive oil. Then mix the resulting liquids completely to obtain the essential oil of Example 2.

[0040] 3. Weigh 0.01g of Atractylodes macrocephala essential oil and dissolve it in 10g of olive oil. Then mix the resulting liquids completely to obtain the essential oil of Example 3.

[0041] 4. Weigh 0.1g of Atractylodes macrocephala essential oil and 0.025g of borneol, dissolve them in 10g of olive oil, and then mix the resulting liquids completely to obtain the essential oil of Example 4.

[0042] III. Preparation of Coating Agent

[0043] Weigh 2.5g of polyvinyl alcohol, let it swell completely, then heat it in a water bath to dissolve it completely. Weigh 1g of Atractylodes macrocephala essential oil, stir it evenly, then add 4ml of glycerin and stir it evenly. Dilute it with distilled water to 100ml to obtain the coating agent.

[0044] IV. Essential Oil Composition Analysis

[0045] The composition of characteristic components of Atractylodes macrocephala essential oil in Example 1 was analyzed by GC-MS and HPLC.

[0046] GC-MS column conditions: SE-30 flexible quartz capillary column (30m × 0.25mm × 0.25μm), helium as carrier gas, temperature program: initial temperature 60℃, hold for 2 min, ramp to 200℃ at a rate of 7℃ / min, then ramp to 250℃ at a rate of 5℃ / min, hold for 3 min; injector temperature 260℃, detector temperature 250℃, split ratio 40:1, injection volume 1μl. Mass spectrometry conditions: ionization source: electron impact ionization (EI), ion source temperature: 230℃, interface temperature: 280℃, electron energy: 70eV.

[0047] HPLC chromatographic conditions: A Waters Sun Fire C18 (250 mm × 4.6 mm, 5 μm) column was used at 30 °C. The injection volume was 20 μL, the flow rate was 1 mL / min, and the injection time was 50 min. The monosaccharide detection wavelengths were 220 nm and 276 nm. The mobile phase was acetonitrile (A)-water (B), with gradient elution: 0–30 min, 50%–70% A; 31–45 min, 78–95% A; 46–50 min, 50% A. System suitability tests were performed, and the results showed that the precision, repeatability, and stability all met the requirements.

[0048] The results showed that the Atractylodes macrocephala essential oil contained characteristic components of Atractylodes macrocephala in Example 1, including atractylodes macrocephala lactone I, atractylodes macrocephala lactone II, atractylodes macrocephala lactone III, atractylone, β-elemene, hesperene, and isohesperene. The relative contents of the above characteristic components are shown in Table 2 below.

[0049] Table 2

[0050] Serial Number Molecular formula molecular weight Compound Name Relative content (%) 1 <![CDATA[C 15 H 18 O2]]> 230 Atractylodes lactone I 0.68 2 <![CDATA[C 15 H 20 O2]]> 232 Atractylodes lactone II 0.75 3 <![CDATA[C 15 H 20 O3]]> 248 Atractylodes lactone III 0.65 4 <![CDATA[C 15 H 20 O]]> 216 Atractylone 41.23 5 <![CDATA[C 15 H 24 ]]> 204 β-elemene 0.37 6 <![CDATA[C 15 H 24 ]]> 204 hesperene 0.28 7 <![CDATA[C 15 H 24 ]]> 204 Isorhosonene 0.43

[0051] The characteristic components in the essential oils and coatings in Examples 2-4 also include: atractylodes lactone I, atractylodes lactone II, atractylodes lactone III, atractylone, β-elemene, hesperidin and isohesperidin.

[0052] V. Comparative Preparation

[0053] Weigh 100g of Atractylodes macrocephala raw material, crush it, pass it through a 60-mesh sieve, add 8 times the amount of water and soak for 3 hours, then bring it to a boil over high heat, turn to low heat and simmer for 2 hours, filter and extract the first decoction; add the same volume of water as the first decoction to the original residue, and extract the decoction a second time in the same way; combine the two extracts, concentrate and dry them for later use.

[0054] Take the extract from the above comparative example, add water to prepare a 20 mg / ml solution for later use.

[0055] VI. Application Example 1

[0056] This invention relates to the therapeutic effect of Atractylodes macrocephala essential oil and its preparations on a mouse model of alopecia areata.

[0057] 1. Laboratory animals

[0058] Fifty SPF-grade male ICR mice, weighing 20-25g, were provided by Zhejiang Academy of Medical Sciences.

[0059] 2. Test drugs and materials

[0060] Example essential oil 1, Example essential oil 2, Example essential oil 3, Example essential oil 4, Comparative decoction, Coating agent, Coating agent excipient group, Positive control.

[0061] The preparation method of the coating agent is as follows: Weigh 2.5g of polyvinyl alcohol, allow it to swell completely, and then dissolve it completely by heating in a water bath. Weigh 1g of Atractylodes macrocephala essential oil, stir evenly, add 4ml of glycerin, stir evenly, and dilute with distilled water to 100ml to obtain the coating agent. Take the above excipients without essential oil and prepare a blank excipient coating solution in the same way.

[0062] Positive control: Minoxidil tincture, Zhejiang Wansheng Pharmaceutical Co., Ltd.

[0063] 3. Modeling

[0064] Experimental mice were randomly divided into four groups: essential oil 1, essential oil 2, essential oil 3, essential oil 4, comparative decoction, film-forming agent, film-forming agent excipient group, positive control, negative control group, and blank control group, with five mice in each group. In the experimental groups, a smooth, hairless area was cleaned on the back of each mouse using depilatory cream. An ointment containing 5% imiquimod was then applied to the back using a cotton swab, ensuring a thin, transparent layer on the untreated area. This was done once daily for two weeks. The blank control group received no treatment. During the modeling process, mice had free access to food and water without any other treatment.

[0065] 4. Experimental Methods

[0066] Mice from all groups of essential oils in Examples 1-4 were placed in IVC independent ventilation cages. Each mouse was swabbed with essential oil 1-4 from Examples and placed in the air inlet of the corresponding ventilation cage to achieve the effect of drug administration by smell. This was done for 14 consecutive days.

[0067] The amount of coating agent used is 1g per piece, for 14 consecutive days.

[0068] The amount of blank excipient coating solution used was 1g / piece, for 14 consecutive days.

[0069] Mice in the comparative decoction group were administered 120 mg / kg by gavage for 14 consecutive days.

[0070] Mice in the positive control group were treated with 0.02 ml of 5% minoxidil solution for 14 consecutive days.

[0071] 5. Statistical methods

[0072] SPSS 26 statistical software was used. Analysis of variance was employed for comparisons between groups, with P < 0.05 considered statistically significant. Quantitative data were expressed as mean ± standard deviation.

[0073] 6. Experimental Results

[0074] 6.1 Gross Quality Score

[0075] During drug administration, the hair growth and quality in the bald areas on the backs of mice were observed, and the growth was graded from 1 to 5. Grade 1: No obvious hair, smooth pink skin; Grade 2: Transparent or semi-transparent stubble at the edge to the center of the bald area; Grade 3: Obvious but short, white hair growing in the bald area; Grade 4: The grown hair is slightly shorter than in the unbald area, and the color is similar; Grade 5: No difference from before hair removal. For ease of statistical analysis, scores were calculated and results compared using the following algorithm: Grades 1-5 were respectively recorded as 1-5 points, and the hair quality score was the total score for each group.

[0076] Scores: Blank control group > Example 4 essential oil group > Example 1 essential oil group > Example 2 essential oil group = coating agent group > Example 3 essential oil group > positive control group > comparative decoction = negative control group = coating agent excipient group. The results are shown in Table 3.

[0077] Table 3

[0078] Grouping Level 1 Level 2 Level 3 Level 4 Level 5 Score Blank control group 0 0 0 0 5 25 Positive control group 0 1 3 1 0 15 negative control group 0 4 1 0 0 11 Example 1: Essential oil set 0 0 1 4 0 19 Example 2: Essential oils 0 0 3 2 0 17 Example 3: Essential oils 0 0 4 1 0 16 Example 4: Essential oils 0 0 1 3 1 20 Comparative water decoction group 0 4 1 0 0 11 Coating agent group 0 0 3 2 0 17 Coating agent excipients group 0 4 1 0 0 11

[0079] The results showed that Atractylodes macrocephala essential oil had a positive effect on hair growth and quality. The essential oils in Examples 1-4 all promoted hair growth and improved hair quality; the Atractylodes macrocephala essential oil coating also had the same effect. However, the comparative water decoction group and the coating excipient group had no significant positive effect on hair growth and quality in mice.

[0080] 6.2 Analysis of Gross Weight and Number of Hair Roots

[0081] A 1×2cm sample skin was randomly selected from the hair removal area, and the hair was removed with a razor. The sample skin was weighed before and after shaving using a balance with an accuracy of 0.01%. The hair weight was obtained by subtracting the skin weight from the hair weight. The hair roots were observed under a 64x stereomicroscope after shaving. A random field of view was photographed using a mobile phone camera at 4.3x magnification, and the number of hair roots in the field of view was counted.

[0082] Gross weight: Blank control group > Example 4 essential oil group > Coating agent group > Example 1 essential oil group > Positive control group > Example 2 essential oil group > Example 3 essential oil group > Comparative decoction > Coating agent excipient group > Negative control group. The results are shown in Table 4.

[0083] Hair root count: Blank control group > Example 4 essential oil group > Coating agent group > Example 1 essential oil group > Example 2 essential oil group > Example 3 essential oil group > Positive control group > Coating agent excipient group > Comparative decoction > Negative control group. The results are shown in Table 4.

[0084] Table 4

[0085] Group Gross weight (mg) Number of hair roots (pieces) Blank control group 56.57±10.46▲▲ 155±41▲▲ Positive control group 38.30±5.33＊▲ 74±12＊＊▲▲ negative control group 18.25±4.54＊＊ 42±18＊＊ Example 1: Essential oil set 45.90±7.80▲▲ 94±28＊＊▲▲ Example 2: Essential oils 38.18±4.92＊▲▲ 81±31＊＊▲▲ Example 3: Essential oils 32.10±9.12＊▲▲ 75±42＊＊▲▲ Example 4: Essential oils 51.18±5.37▲▲ 119±25＊＊▲▲ Comparative water decoction group 19.32±3.80＊＊ 41±37＊＊ Coating agent group 48.89±4.42▲▲ 108±29＊＊▲▲ Coating agent excipients group 19.42±3.62＊＊ 51±22＊＊

[0086] Note: Compared with the CK group: *P<0.05, **P<0.01; Compared with the negative group: ▲P<0.05, ▲▲P<0.01.

[0087] The results showed that Atractylodes macrocephala essential oil had a positive effect on hair growth and hair root recovery. The Atractylodes macrocephala essential oil in Examples 1-4 promoted hair growth and hair root recovery, and the Atractylodes macrocephala essential oil coating also had the same effect. However, the comparative decoction group, the coating excipient group, and the comparative decoction group had no significant positive effect on hair growth and hair root recovery in mice.

[0088] VII. Application Example 2

[0089] This invention relates to the therapeutic effect of Atractylodes macrocephala essential oil and its preparations on a mouse model of hair loss.

[0090] 1. Laboratory animals

[0091] Fifty SPF-grade male ICR mice, weighing 20-25g, were provided by Zhejiang Academy of Medical Sciences.

[0092] 2. Test drugs and materials

[0093] Essential oil of Example 1, essential oil of Example 2, essential oil of Example 3, essential oil of Example 4, comparative decoction, coating agent, coating agent excipient group, positive control.

[0094] The preparation method of the coating agent is as follows: Weigh 2.5g of polyvinyl alcohol, allow it to swell completely, and then dissolve it completely by heating in a water bath. Weigh 1g of Atractylodes macrocephala essential oil, stir evenly, add 4ml of glycerin, stir evenly, and dilute with distilled water to 100ml to obtain the coating agent. Take the above excipients without essential oil and prepare a blank excipient coating solution in the same way.

[0095] Positive control: Minoxidil tincture, Zhejiang Wansheng Pharmaceutical Co., Ltd.

[0096] 3. Modeling

[0097] Experimental mice were randomly divided into four groups: essential oils from Example 1, Example 2, Example 3, and Example 4; a comparative aqueous decoction; a coating agent; a coating agent excipient group; a positive control group; a negative control group; and a blank control group, with five mice in each group. During the modeling process, mice were allowed free access to food and water without any other treatment.

[0098] Mice in the experimental group underwent hair removal treatment, with the hair removal area measuring 2cm × 3cm, using an animal-specific electric hair removal device and animal-specific hair removal solution. The mice were then weighed and marked. (Marking: Picric acid was used to mark the backs of the mice, and markings were also made on the head, left forelimb, right forelimb, left hindlimb, and right hindlimb.) Except for the control group, all other groups received subcutaneous injections of testosterone propionate for 14 consecutive days to maintain high androgen levels in the mice.

[0099] The testosterone propionate injection used in this experiment was at a concentration of 25 mg / ml. If the original concentration of testosterone propionate injection were used for direct subcutaneous injection into mice, the daily dose would be 0.004 ml, which would be difficult to control and could easily lead to significant errors. To reduce errors and improve the reliability of the experiment, the testosterone propionate injection was diluted with injectable grade soybean oil before injection. Following a dilution ratio of 4:1, 80 ml of injectable grade soybean oil and 20 ml of testosterone propionate were measured, mixed thoroughly, and shaken to obtain a 5 mg / ml testosterone propionate injection solution. This solution was then sealed and stored away from light for later use.

[0100] 4. Experimental Methods

[0101] Thirty minutes after the injection of testosterone propionate, the negative control group was treated with injectable soybean oil.

[0102] The positive control group was treated with 0.02 ml of 5% minoxidil solution.

[0103] Mice from all groups of essential oils in Examples 1-4 were placed in IVC independent ventilation cages. Each mouse was swabbed with essential oil 1-4 from Examples and placed in the air inlet of the corresponding ventilation cage to achieve the effect of drug administration by smell. This was done for 14 consecutive days.

[0104] The amount of coating agent used is 1g per piece, for 14 consecutive days.

[0105] The amount of blank excipient coating solution used was 1g / piece, for 14 consecutive days.

[0106] Mice in the comparative decoction group were administered 120 mg / kg by gavage for 14 consecutive days.

[0107] 5. Statistical methods

[0108] SPSS 26 statistical software was used. Analysis of variance was employed for comparisons between groups, with P < 0.05 considered statistically significant. Quantitative data were expressed as mean ± standard deviation.

[0109] 6. Experimental Results

[0110] 6.1 Gross Quality Score

[0111] During drug administration, the hair growth and quality in the bald areas on the backs of mice were observed, and the growth was graded from 1 to 5. Grade 1: No obvious hair, smooth pink skin; Grade 2: Transparent or semi-transparent stubble at the edges to the center of the bald area; Grade 3: Obvious but short, white hair growing in the bald area; Grade 4: The grown hair is slightly shorter than in the unbald area, and the color is similar; Grade 5: No difference from before hair removal. For ease of statistical analysis, scores were calculated and results compared using the following algorithm: Grades 1-5 were respectively recorded as 1-5 points, and the hair quality score was the total score for each group.

[0112] Scores: Blank control group > Example essential oil group 4 > Example essential oil group 1 = Coating agent group > Positive control group > Example essential oil group 2 > Example essential oil group 3 > Comparative decoction = Negative control group = Coating agent excipient group. The results are shown in Table 5.

[0113] Table 5

[0114] Grouping Level 1 Level 2 Level 3 Level 4 Level 5 Score Blank control group 0 0 0 0 5 25 Positive control group 0 0 2 3 0 18 negative control group 0 4 1 0 0 11 Example 1: Essential Oil Set 0 0 1 4 0 19 Example 2: Essential Oil Set 0 0 3 2 0 17 Example 3: Essential Oil Set 0 0 4 1 0 16 Example 4: Essential Oil Set 0 0 1 2 2 21 Comparative water decoction group 0 4 1 0 0 11 Coating agent group 0 0 1 4 0 19 Coating agent excipients group 0 4 1 0 0 11

[0115] The results showed that Atractylodes macrocephala essential oil had a positive effect on hair growth and quality. Specifically, the essential oils in Examples 1-4 all promoted hair growth and improved hair quality, and the Atractylodes macrocephala essential oil coating also had the same effect. However, the comparative decoction group, the coating excipient group, and the comparative decoction group had no significant positive effect on hair growth and quality in mice.

[0116] 6.2 Analysis of Gross Weight and Number of Hair Roots

[0117] One day before euthanasia, the male pattern baldness mouse model was given a final injection, followed by a 12-hour fast for each group of mice. Final photographs were taken, and the mice's hair growth, activity level, and weight were observed and recorded. Mice were euthanized by dislocation, skinned, and the designated hair-removed area was cut off, sealed, and refrigerated for later use. The mice were dissected, and the internal organs were removed, weighed, and photographed. After dissection, the sealed, refrigerated skin sac was removed, and the weight of the unremoved hair was measured using a precision electronic balance. Then, the hair in the experimental area was scraped off using a specialized hair removal instrument, ensuring cleanliness and no damage. The cleanly scraped skin sac was then weighed using a precision electronic balance to obtain the hair weight. Finally, the number of hair follicles was observed under a stereomicroscope at 4x magnification, and photographs were taken and recorded.

[0118] Gross weight: Blank control group > Example 4 essential oil group > Coating agent group > Example 1 essential oil group > Positive control group > Example 2 essential oil group > Example 3 essential oil group > Negative control group > Coating agent excipient group > Comparative decoction. The results are shown in Table 6.

[0119] Hair root count: Blank control group > Example 4 essential oil group > Coating agent group > Example 1 essential oil group > Example 2 essential oil group > Positive control group > Example 3 essential oil group > Coating agent excipient group > Comparative decoction > Negative control group. The results are shown in Table 6.

[0120] Table 6

[0121] Group Gross weight (mg) Number of hair roots (pieces) Blank control group 62.57±8.53▲▲ 146±36▲▲ Positive control group 42.30±3.42*▲ 73±14**▲▲ negative control group 20.43±5.63** 39±11** Example 1: Essential Oil Set 44.63±4.73▲▲ 100±22**▲▲ Example 2: Essential Oil Set 41.24±5.32*▲▲ 85±32**▲▲ Example 3: Essential Oil Set 38.46±5.37*▲▲ 72±25**▲▲ Example 4: Essential Oil Set 51.11±6.27▲▲ 114±16**▲▲ Comparative water decoction group 19.72±2.81** 40±17** Coating agent group 49.82±7.12▲▲ 106±23**▲▲ Coating agent excipients group 20.12±4.12** 50±21**

[0122] Note: Compared with the CK group: *P<0.05, **P<0.01; Compared with the negative group: ▲P<0.05, ▲▲P<0.01.

[0123] The results showed that the essential oil from the Atractylodes macrocephala example promoted hair growth and hair root recovery, and the Atractylodes macrocephala essential oil coating also had the same effect. However, the comparative decoction group, the coating excipient group, and the comparative decoction group did not show significant effects in promoting hair growth and hair root recovery.

[0124] Eight Application Example 3

[0125] This invention relates to a human clinical observation of the therapeutic effects of Atractylodes macrocephala essential oil and its preparations on alopecia areata and hair loss.

[0126] 1. General Information

[0127] One hundred patients each of alopecia areata and alopecia areata were recruited and randomly divided into four groups. The treatment group consisted of 50 patients with alopecia areata and 50 patients with alopecia areata, aged 17-51 years. The control group consisted of 50 patients with alopecia areata and 50 patients with alopecia areata, aged 20-50 years. The two groups had similar age and gender distributions (P>0.05) and were comparable.

[0128] Inclusion criteria for patients with alopecia areata: meeting the above diagnostic criteria for alopecia areata; aged 16-53 years; good compliance; no history of diabetes, cardiovascular disease, malignant tumors, hematological diseases, etc.; patients understood the purpose of this study, voluntarily participated, and gave their consent. Inclusion criteria for patients with hair loss: clinically diagnosed with seborrheic alopecia; outpatients aged 23-51 years; had not used glucocorticoids or immunosuppressants within one month prior to treatment; and had not taken related traditional Chinese medicine or androgen-suppressing drugs within two weeks.

[0129] 2 Treatment methods

[0130] The treatment group received an external application of Atractylodes macrocephala essential oil film. Patients applied the film to the affected area with a soft brush or cotton swab, ensuring the solution covered the entire area. Gently massage the area until a slight warming sensation is felt. This was done twice daily. The control group received a 3% minoxidil hair growth solution. No other hair growth therapies were used during the treatment period.

[0131] 3. Efficacy criteria

[0132] 3.1 Criteria for Evaluating the Efficacy of Alopecia Areata: Based on the "Guiding Principles for Clinical Research of New Traditional Chinese Medicines, Volume 3" regarding the evaluation of the efficacy of alopecia areata, the criteria are as follows: Clinically Cured: Hair loss stops, all bald hair grows back, and its distribution density, color, and thickness are the same as the healthy hair area, and sebum secretion returns to normal; Significantly Effective: Hair loss stops, hair regeneration reaches more than 70%, and its density, color, and thickness are close to the healthy hair area, and sebum secretion is significantly reduced; Effective: Hair loss stops, hair regeneration reaches more than 30%, and vellus hair and blood hair grow back; Ineffective: Hair regeneration is less than 30% or hair loss continues.

[0133] 3.2 Criteria for judging the efficacy of hair loss treatment: Cured: Hair loss stops, there is no itching, there is basically no oily feeling, new hair or vellus hair appears in the lesion area, and all the hair in the bald patch grows back and is terminal hair; Significantly effective: Vellus hair is generally present in the bald patch, and terminal hair accounts for more than 50% of the bald patch area; Ineffective: No new hair growth, no reduction in oiliness and sebaceous scaling, no relief from itching, only vellus hair in the bald patch, no or few terminal hairs seen, or the bald patch area continues to expand.

[0134] 4 Statistical Methods

[0135] SPSS 26 statistical software was used. Analysis of variance was used for comparisons between groups. P < 0.05 was considered statistically significant.

[0136] 5. Treatment Results

[0137] The patients' alopecia areata improved significantly. The treatment group showed significantly better results than the control group. Moreover, this formula was more effective than topical 3% minoxidil hair growth solution in treating alopecia areata. The results are shown in Table 7 and are worthy of clinical application.

[0138] The results of the subjects' use of the drug showed that the topical application of Atractylodes macrocephala essential oil film can promote local blood circulation, improve hair nutrition, promote hair growth and drug absorption, so it can achieve a good effect of hair growth and hair strengthening. The patients' hair loss was significantly improved. The results are shown in Table 8.

[0139] Table 7

[0140] Group get well Effective efficient invalid Efficiency / % Treatment group 8 18 9 15 70 control group 5 17 10 18 64

[0141] Table 8

[0142] Group get well Effective efficient invalid Efficiency / % Treatment group 6 12 15 17 66 control group 4 8 16 22 56

[0143] Application Example 4

[0144] This application example relates to the preparation of an essential oil patch for hair loss. The preparation process is as follows: 500g of Atractylodes macrocephala herb is mixed, pulverized, and extracted with the following herbs:

[0145] A: 60g dried ginger medicinal material

[0146] B: 60g fennel seeds

[0147] C: 60g of mugwort herbs

[0148] D: 60g cinnamon herbs

[0149] The above-mentioned herbal powders were placed separately into extraction vessels, and essential oil mixtures were obtained by subcritical extraction for later use. The subcritical extraction conditions used were: a solid-liquid ratio of 1:9 (m:v), an extraction temperature of 50℃, a pressure of 0.9MPa, an extraction time of 6h, and butane as the solvent. The mixtures were separated twice, combined, and labeled as essential oil mixtures A, B, C, and D, respectively, for later use.

[0150] The above essential oil mixture was analyzed by GC-MS and HPLC and found to contain the following characteristic components: atractylodes lactone I, atractylodes lactone II, atractylodes lactone III, atractylone, β-elemene, hesperidin and isohesperidin.

[0151] Weigh PEG 400 and PEG 4000 and essential oils A, B, C, and D respectively in a 4:3:3 ratio, heat and mix thoroughly, then cool to form a viscous semi-solid fluid. Take an appropriate amount of the semi-solid fluid and evenly coat it onto an aluminum foil backing layer. Let it cure at room temperature for 5-10 minutes to allow the solvent to fully evaporate. Cover with an EVA controlled-release film, heat-seal, attach medical double-sided tape, cut to the appropriate size according to usage requirements, and seal in packaging to obtain patches A, B, C, and D. Prepare patch E separately using the same method with Atractylodes macrocephala essential oil.

[0152] Therapeutic effect observation: 50 patients with hair loss were randomly divided into 5 groups. Patch A, Patch B, Patch C, Patch D, and Patch E.

[0153] The administration method was as follows: Patches A, B, C, D, and E were applied to the Shenque acupoint for 4 hours daily for 60 consecutive days. The therapeutic effects and indicators were observed using the same methods as in Example 3. The results are shown in Table 9.

[0154] Results: The patch, through skin absorption of essential oils, can achieve hair growth, hair loss prevention, and hair loss improvement effects comparable to the essential oil in Example 14, and patches A through D all have a certain synergistic effect.

[0155] Table 9

[0156] Group get well Effective efficient invalid Efficiency / % Patch A 2 4 1 3 70 Patch B 1 3 3 3 70 Patch C 0 3 4 3 70 Patch D 2 4 1 3 70 Patch E 0 2 4 4 60

[0157] Those skilled in the art should understand that the discussion of any of the above embodiments is merely exemplary and is not intended to imply that the scope of this disclosure (including the claims) is limited to these examples; within the framework of this disclosure, the technical features of the above embodiments or different embodiments can also be combined, the steps can be implemented in any order, and there are many other variations of different aspects of one or more embodiments of this specification as described above, which are not provided in detail for the sake of brevity.

[0158] One or more embodiments of this specification are intended to cover all such substitutions, modifications, and variations that fall within the broad scope of the appended claims. Therefore, any omissions, modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of one or more embodiments of this specification should be included within the scope of protection of this disclosure.

Claims

1. The use of Atractylodes macrocephala essential oil in the preparation of medicines for the prevention and treatment of hair loss and alopecia areata, characterized in that, The Atractylodes macrocephala essential oil comprises a water-insoluble volatile oil obtained by extracting the dried tuberous roots of the traditional Chinese medicine Atractylodes macrocephala Koidz. and its cultivated varieties through supercritical CO2 extraction, subcritical fluid extraction, or steam distillation; wherein, The supercritical CO2 extraction process conditions include: extraction pressure of 15-30 MPa, extraction temperature of 30-50°C, extraction time of 2-4.5 hours, and CO2 flow rate of 15-40 L / h. The subcritical fluid extraction process conditions include: solvent addition of 5 to 10 times the amount of raw material, temperature of 35 to 55°C, pressure of 0.3 to 0.8 MPa, and extraction time of 1 to 8 hours; the solvent is butane. The process conditions for steam distillation include: soaking for 2.5 to 6 hours, followed by distillation for 5 to 9 hours; The characteristic components of the Atractylodes macrocephala essential oil include: atractylodes macrocephala lactone I, atractylodes macrocephala lactone II, atractylodes macrocephala lactone III, atractylone, β-elemene, hesperene and isohesperene.

2. The use according to claim 1, characterized in that, The drug includes an Atractylodes macrocephala essential oil complex obtained by mixing the Atractylodes macrocephala essential oil with traditional Chinese medicine or traditional Chinese medicine essential oil in a certain proportion, or an Atractylodes macrocephala essential oil complex obtained by compounding the Atractylodes macrocephala essential oil, traditional Chinese medicine essential oil and organic solvent.

3. The use according to claim 2, characterized in that, The herbal essential oils include essential oils from at least one of dried ginger, fennel, mugwort, and cinnamon.

4. The use according to claim 1, characterized in that, The dosage form of the drug includes at least one of nasal administration and transdermal administration.

5. The use according to claim 4, characterized in that, The dosage form includes at least one of fumigants, inhalers, sprays, aerosols, ointments, patches, liniments, tinctures, lotions, oils, films, drops, and suppositories.

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