Use of Chinese fir oil in preparation of medicine for promoting hair growth and relieving alopecia

Abstract

The application provides application of Chinese fir oil in preparation of a medicine for promoting hair growth and relieving alopecia, and belongs to the technical field of application of Chinese fir oil. The application finds that the Chinese fir oil can promote hair growth of normal alopecia and alopecia mice caused by injury, is safe in use, and can be used alone or combined with traditional Chinese medicinal materials or chemical medicines having a hair-growing effect, and is used for treating alopecia areata, seborrheic alopecia, chemotherapy-induced alopecia, senile alopecia or chronic telogen effluvium, and meets the demand of people for anti-hair-loss products.

Description

[0001] This application relies on and claims priority to Chinese Patent Application No. 202311036678.0, filed on August 17, 2023, the entire contents of which are incorporated herein by reference. Technical Field

[0002] This invention relates to the field of cedar oil application technology, and more particularly to the application of cedar oil in the preparation of drugs that promote hair growth and relieve hair loss. Background Technology

[0003] Hair loss, when it occurs on the scalp, is also known as baldness or alopecia. While hair loss can occur on any part of the body, scalp hair loss is a cosmetic issue that receives considerable public attention. Common types and causes of scalp hair loss include: androgenetic alopecia, alopecia areata, systemic diseases (such as lupus erythematosus and hormone-related diseases), medications (chemotherapy drugs), infections (such as fungal infections); physical stress (such as high fever, surgery, serious illness, sudden weight loss, pregnancy), psychological stress, and skin injuries (such as trauma).

[0004] Hair loss can be treated according to its underlying cause. Common treatments include medication, such as minoxidil and finasteride, and surgery, such as hair transplantation. However, minoxidil and finasteride have significant side effects. Finasteride carries a potential risk of external genital malformations in male fetuses and is therefore contraindicated in pregnant women or women of childbearing age. Minoxidil has a short-lived effect and several adverse reactions, such as itching and dermatitis, have been reported. Surgical treatment is expensive, and postoperative patient adherence is low. Traditional Chinese medicine, on the other hand, is adaptable, has fewer side effects, is readily available, and is low-cost. Studies have shown that many traditional Chinese medicines have the effect of preventing hair loss and promoting hair growth.

[0005] For example, patent CN111603495A discloses a solution of Ligusticum chuanxiong plant extract and its application in the preparation of drugs that promote hair growth. The Ligusticum chuanxiong plant extract is used in hair growth research. Specifically, it can promote the proliferation of human dermal papilla cells, increase the number of hair follicle cells, promote hair follicle growth, increase hair length, and achieve the effect of hair growth.

[0006] For example, patent CN101628047A discloses a traditional Chinese medicine for treating hair loss. The medicine is mainly composed of fresh gardenia, fresh walnut, and fresh arborvitae leaves. Specifically, the fresh gardenia, fresh walnut, and fresh arborvitae leaves are mashed into a "mud" with winter snow water and then soaked in winter snow water for two hours before being used to wash the hair. After 10 consecutive days of use, clinical use has shown that patients can relieve symptoms during use and prevent abnormal hair loss after use.

[0007] Cunninghamia lanceolata (Lamb.) Hook, a plant of the Taxaceae family, is a yellow, semi-transparent oily liquid obtained through methods such as steam extraction, dry distillation, and supercritical fluid extraction. Representative components of cedar oil include α-pinene, α-juniperene, β-juniperene, thujone, and juniper borneol. Currently, cedar oil is mainly used for fragrance; it has not yet been used to prevent hair loss or promote hair regeneration. However, it has been found to have the effect of halting hair loss and promoting hair regeneration with low side effects, showing good development prospects and commercial value. This invention provides a basis for developing safe, efficient, and inexpensive hair growth drugs and products. Summary of the Invention

[0008] The purpose of this invention is to address the shortcomings of existing technologies by providing an application of cedar oil in the preparation of drugs that promote hair growth and alleviate hair loss. This drug is effective in preventing hair loss and promoting hair regeneration for people with alopecia areata, seborrheic alopecia, chemotherapy-induced alopecia, senile alopecia, and chronic telogen effluvium, with low side effects and good development prospects and commercial value.

[0009] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0010] The application of cedar oil in the preparation of drugs that promote hair growth and relieve hair loss, wherein the hair loss is alopecia areata, seborrheic alopecia, chemotherapy-induced alopecia, senile alopecia, and chronic telogen effluvium.

[0011] Furthermore, the cedar oil is prepared from cedar wood using conventional extraction and separation methods.

[0012] Preferably, the extraction method includes, but is not limited to, steam extraction, dry distillation or supercritical CO2 extraction; the separation method includes, but is not limited to, fractionation, crystallization, freezing and various chromatographic separation methods.

[0013] Preferably, the method for preparing the cedar oil includes the following steps:

[0014] The stems, branches, roots, and trunks of Chinese fir are crushed and put into an extraction vessel. Microwave-assisted supercritical CO2 extraction is used to obtain Chinese fir oil.

[0015] More preferably, the particle size of the pulverized material is 3-50 mm.

[0016] More preferably, during the extraction process, the extraction temperature is 30-45℃, the CO2 temperature is 25-75℃, the CO2 flow rate is 1-10L / h, the extraction pressure is 15-55Mpa, and the extraction time is 1-8h.

[0017] More preferably, during the supercritical CO2 extraction process, microwave heating is used, and the microwave is intermittently activated to maintain the temperature.

[0018] Furthermore, the cedar oil, by weight percentage, contains the following components: 18-70% α-cedrol, 1-25% α-cedrolene, 0-11% β-cedrolene, 0-5% α-pinene, and 1-5% cypressene.

[0019] Furthermore, the cedar oil can be used alone or in combination with traditional Chinese medicinal materials or chemical drugs that have hair growth effects.

[0020] Preferably, the medicinal materials include at least one of the following: cypress seed, chuanxiong rhizome, gardenia fruit, ginger, ginseng, lingzhi mushroom, rhubarb, he shou wu, Sichuan pepper, pinellia tuber, psoralea fruit, black sesame, gastrodia elata, cornus fruit, arborvitae leaf, mint, artemisia argyi, saposhnikovia root, vitex fruit, wolfberry, safflower, angelica sinensis, salvia miltiorrhiza, citron, astragalus root, privet fruit, hollyhock fruit, wild hawthorn fruit, polyporus umbellatus, sandalwood, ginkgo, chuanxiong rhizome, sanguisorba officinalis, and prepared rehmannia root.

[0021] Preferably, the chemical drug includes at least one of minoxidil, finasteride, α-estradiol, stilbene glycoside, cyclosporine, anthraquinone, pilocarpine, carboplatin chloride, and cimetidine.

[0022] Furthermore, the cedar oil is formulated into topical or oral preparations with a pharmaceutically acceptable carrier.

[0023] Specifically, the drug may be a topical preparation or an oral preparation; the topical preparation may be any one of shampoo, hair dye, hair conditioner, lotion, serum, ointment, gel, patch, tincture or spray, and the oral preparation may be any one of tablet, capsule, granule, pill or oral liquid.

[0024] Specifically, the carrier used is selected from at least one of the conventional fillers, binders, disintegrants, lubricants, solubilizers, penetration enhancers, suspending agents, wetting agents, pigments, fragrances, solvents, surfactants, or flavoring agents in the pharmaceutical field; including but not limited to lactose, kaolin, sucrose, talc, gelatin, agar, pectin, gum arabic, magnesium stearate, stearic acid, syrup, peanut oil, olive oil, water, and ethanol.

[0025] Compared with the prior art, the present invention has the following beneficial effects:

[0026] The cedar oil of this invention has the effect of promoting hair growth and relieving hair loss, which increases the new use of cedar oil and has industrial utilization value. Moreover, the results can be applied to medical clinical practice to promote hair growth, and to the development of hair growth drugs and cosmetics, which has potential economic and social benefits and meets people's demand for hair loss prevention products. Detailed Implementation

[0027] The following non-limiting embodiments are intended to enable those skilled in the art to gain a more comprehensive understanding of the present invention, but do not limit the invention in any way. The following description is merely an exemplary illustration of the scope of protection of the present invention, and those skilled in the art can make various changes and modifications to the invention based on the disclosed content, which should also fall within the scope of protection of the present invention.

[0028] In the following embodiments, the cedar oil was prepared by the following method:

[0029] 10 kg of Chinese fir root material was crushed and passed through a 20-mesh sieve, then added to an extraction vessel and microwaved to 53°C. Supercritical CO2 was introduced, and the extraction pressure was 28 MPa, temperature 43°C, time 3.5 h, and flow rate 10 kg CO2 / (h·kg). Microwave heating was intermittently started for 5 min every 30 min. The extracted extract was then separated in a separation vessel under reduced pressure at 10 MPa and temperature 35°C. The Chinese fir oil was collected, with a yield of 0.98%. GC-MS analysis revealed the following contents in the Chinese fir oil: α-cedrol 68%, α-juniperene 7%, β-juniperene 2%, α-pinene 1%, and juniperene 2%.

[0030] The present invention will be further described below by way of specific embodiments. Unless otherwise specified, all chemical reagents used in the embodiments of the present invention are obtained through conventional commercial means.

[0031] Example 1: Functional experiment of cedar oil promoting hair growth in normal C57BL / 6 mice

[0032] 1. Experimental drugs

[0033] 95% ethanol was purchased from Sinopharm Chemical Reagent Co., Ltd.; 5% minoxidil liniment was purchased from Shandong Jingwei Pharmaceutical Co., Ltd., with the national drug approval number H20233798.

[0034] 2. Laboratory animals

[0035] C57BL / 6 mice, SPF grade, 6-8 weeks old, were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd.

[0036] 3. Experimental Grouping and Experimental Design

[0037] Mice were lightly anesthetized with ether, and their backs were shaved using a mixture of rosin and paraffin in equal proportions, covering an area of ​​approximately 2cm × 3cm. The shaved mice were then randomly divided into 5 groups of 20 mice each, and the administration regimens are shown in Table 1.

[0038] Table 1. Dosing Regimen

[0039] Group Dosing regimen Blank group Apply an equal amount of 95% ethanol evenly Low-dose group of cedar oil Apply an equal amount of 95% ethanol solution of cedar oil evenly, with a cedar oil content of 20 mg / kg. Medium dose group of cedar oil Apply an equal amount of 95% ethanol solution of cedar oil evenly, with a cedar oil content of 40 mg / kg. High-dose group of cedar oil Apply an equal amount of 95% ethanol solution of cedar oil evenly, with a cedar oil content of 80 mg / kg. Positive control group Apply an equal amount of 5% minoxidil lotion evenly.

[0040] After hair removal, mice were treated with 1 mL of the medication twice daily for 18 consecutive days. Hair growth in the bald areas was recorded, and mice were sacrificed on day 19. Hair from the bald areas was collected and its length and weight were measured. At the end of the experiment, skin from the bald areas on the backs of the mice was taken, fixed in formalin, and prepared for pathological sectioning to observe hair follicle growth.

[0041] 4. Experimental Results

[0042] 4.1 Hair growth status

[0043] After hair removal, the skin on the backs of mice appeared pink. In the cedar oil group, the hair-removed areas began to turn gray on days 2-3, and were covered with gray downy hair on days 5-6. By days 8-9, the hair-removed areas were almost completely covered with hair. After 18 days of continuous administration, compared with the control group, hair growth was significantly increased in the medium- and high-dose groups and the positive control group, while the high-dose group and the positive control group showed essentially the same results.

[0044] 4.2 Number of hair follicles in the growth phase

[0045] At the end of the experiment, skin from the hairless area on the back of the mice was taken, fixed with formalin, and used for pathological sections. After staining, the longitudinal section and the transverse section at the same level of the mouse hair follicles were observed under a microscope. The results are shown in Table 2.

[0046] Table 2. Number of hair follicles in the growth phase

[0047] Group Number of hair follicles in the growth phase (per field of view) Blank group 34±7 Low-dose group of cedar oil 43±10 Medium dose group of cedar oil 58±8* High-dose group of cedar oil 68±13* Positive control group 67±12*

[0048] Note: * indicates that P < 0.05 compared to the control group.

[0049] As shown in the table above, compared with the blank group, the mice in the cypress oil group and the positive control group had a greater number of hair follicles in the anagen phase. Among them, the number of hair follicles in the anagen phase of the medium- and high-dose groups and the positive control group were significantly different from those in the blank group. The number of hair follicles in the anagen phase of the high-dose group and the positive control group was comparable, indicating that the activity of the high-dose cypress oil was comparable to that of the positive control drug.

[0050] 4.3 Length and weight of new hair growth

[0051] Mice were sacrificed at 19 days, and the length and weight of hair in the bald areas were measured. The results are shown in Table 3.

[0052] Table 3. Mouse hair length and weight

[0053] Group New hair growth length (mm) <![CDATA[New hair weight (mg / cm 2 )]]> Blank group 6.39±0.35 13.7±2.4 Low-dose group of cedar oil 6.98±0.29 15.1±5.1 Medium dose group of cedar oil 8.63±0.61* 16.8±4.6* High-dose group of cedar oil 8.89±0.28* 17.9±5.8* Positive control group 9.01±0.32* 18.4±4.3*

[0054] Note: * indicates that P < 0.05 compared to the control group.

[0055] As shown in the table above, compared with the blank group, the length and weight of newly grown hair in mice in the medium- and high-dose groups of Chinese fir oil and the positive control group were significantly increased, showing a significant difference from the blank group (P<0.05). The hair growth value of mice in the high-dose group was comparable to that of the positive control group, indicating that the activity of the high-dose group of Chinese fir oil was comparable to that of the positive control drug.

[0056] Example 2: Functional experiment of cedarwood oil promoting hair growth in mice with cyclophosphamide-induced alopecia.

[0057] 1. Experimental drugs

[0058] 95% ethanol was purchased from Sinopharm Chemical Reagent Co., Ltd.; 5% minoxidil liniment was purchased from Shandong Jingwei Pharmaceutical Co., Ltd., National Drug Approval Number H20233798; cyclophosphamide for injection was purchased from Jiangsu Hengrui Medicine Co., Ltd., National Drug Approval Number H20023036.

[0059] 2. Laboratory animals

[0060] C57BL / 6 mice, SPF grade, 6-8 weeks old, were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd.

[0061] 3. Experimental Grouping and Experimental Design

[0062] Mice were lightly anesthetized with ether, and their backs were shaved using a rosin / wax mixture in equal proportions, covering an area of ​​approximately 2cm × 3cm. After shaving, the test drug was applied twice daily to the shaved area. Seven days later, mice were given a single intraperitoneal injection of cyclophosphamide 150 mg / kg. -1 This study aimed to examine the effect of cedarwood oil on promoting hair growth in mice with cyclophosphamide-induced alopecia. After hair removal, the mice were randomly divided into 5 groups of 20 mice each, and the drug administration regimens are shown in Table 4.

[0063] Table 4. Dosing Regimen

[0064] Group Dosing regimen Blank group Apply an equal amount of 95% ethanol evenly Low-dose group of cedar oil Apply an equal amount of 95% ethanol solution of cedar oil evenly, with a cedar oil content of 20 mg / kg. Medium dose group of cedar oil Apply an equal amount of 95% ethanol solution of cedar oil evenly, with a cedar oil content of 40 mg / kg. High-dose group of cedar oil Apply an equal amount of 95% ethanol solution of cedar oil evenly, with a cedar oil content of 80 mg / kg. Positive control group Apply an equal amount of 5% minoxidil lotion evenly.

[0065] After hair removal in mice, apply 1 mL of the medication twice daily to the bald area. After 7 days, administer 150 mg / kg of the medication. -1 Cyclophosphamide was administered to mice for 18 days. Hair growth in the bald areas was recorded, and mice were sacrificed on day 19. Hair from the bald areas was collected and its length and weight were measured. At the end of the experiment, skin from the bald areas on the backs of the mice was taken, fixed with formalin, and prepared for pathological sectioning to observe hair follicle growth.

[0066] 4. Experimental Results

[0067] 4.1 Hair growth status

[0068] After hair removal, the skin on the back of the mice turned pink. On days 2-3, the hair-removed area began to turn gray. On days 5-6, the hair-removed area was covered with gray downy hair. On days 8-9, the hair-removed area was basically covered with hair. After 18 days of continuous administration, compared with the control group, the hair growth of mice in the medium- and high-dose groups and the positive control group was significantly increased. The hair growth of mice in the high-dose group and the positive control group was basically the same.

[0069] 4.2 Number of hair follicles in the growth phase

[0070] At the end of the experiment, skin from the hairless area on the back of the mice was taken, fixed with formalin, and used for pathological sections. After staining, the longitudinal sections and transverse sections of the mouse hair follicles at the same level were observed under a microscope. The results are shown in Table 5.

[0071] Table 5. Number of hair follicles in the growth phase

[0072] Group Number of hair follicles in the growth phase (per field of view) Blank group 24±7 Low-dose group of cedar oil 31±11 Medium dose group of cedar oil 48±10* High-dose group of cedar oil 54±14* Positive control group 52±9*

[0073] Note: * indicates that P < 0.05 compared to the control group.

[0074] As shown in the table above, compared with the blank group, the number of hair follicles in the anagen phase of mice in the cedar oil group and the positive control group was greater. Among them, the number of hair follicles in the anagen phase of mice in the medium and high dose groups and the positive control group was significantly different from that in the blank group (P<0.05). Compared with the positive control group, the number of hair follicles in the anagen phase of mice in the high dose group was slightly higher.

[0075] 4.3 Length and weight of new hair growth

[0076] After 19 days, the length and weight of the hair in the baldened areas were measured. The results are shown in Table 6.

[0077] Table 6. Length and weight of newborn mouse fur

[0078] Group New hair growth length (mm) <![CDATA[New hair weight (mg / cm 2 )]]> Blank group 4.67±0.37 12.1±1.7 Low-dose group of cedar oil 6.09±0.41 14.3±3.1 Medium dose group of cedar oil 7.26±0.52 16.2±3.9* High-dose group of cedar oil 8.19±0.26* 17.2±3.6* Positive control group 8.28±0.38* 17.7±2.9*

[0079] Note: * indicates that P < 0.05 compared to the control group.

[0080] As shown in the table above, compared with the blank group, there were significant differences in the length and weight of newly grown hair in the high-dose group and the positive control group (P<0.05). Compared with the positive control group, the number of hair follicles in the growth phase of the high-dose group was comparable to that of the positive control group.

[0081] Example 3: Experiment on the effect of cedarwood oil in combination with other drugs on hair growth in normal C57BL / 6 mice

[0082] 1. Experimental drugs

[0083] 95% ethanol was purchased from Sinopharm Chemical Reagent Co., Ltd.; 2% minoxidil liniment was purchased from Shandong Jingwei Pharmaceutical Co., Ltd., with national drug approval number H20233798.

[0084] 2. Laboratory animals

[0085] C57BL / 6 mice, SPF grade, 6-8 weeks old, were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd.

[0086] 3. Experimental Grouping and Experimental Design

[0087] Mice were lightly anesthetized with ether, and their backs were shaved using a rosin / wax mixture in equal proportions, covering an area of ​​approximately 2cm × 3cm. After shaving, the mice were randomly divided into 5 groups of 20 mice each, and the administration regimens are shown in Table 7.

[0088] Table 7. Dosing Regimen

[0089]

[0090]

[0091] After hair removal, mice were treated with 1 mL of the medication twice daily for 18 consecutive days. Hair growth in the bald areas was recorded, and mice were sacrificed on day 19. Hair from the bald areas was collected and its length and weight were measured. At the end of the experiment, skin from the bald areas on the backs of the mice was taken, fixed in formalin, and prepared for pathological sectioning to observe hair follicle growth.

[0092] 4. Experimental Results

[0093] 4.1 Hair growth status

[0094] Compared to the control group, the mice in the cedar oil group had denser and better-colored hair in the bald area.

[0095] 4.2 Length and weight of new hair growth

[0096] After 19 days, the length and weight of the hair in the bald areas of the mice were measured. The results are shown in Table 8.

[0097] Table 8. Length and weight of newborn mouse fur

[0098] Group New hair growth length (mm) <![CDATA[New hair weight (mg / cm 2 )]]> Blank group 6.58±0.48 14.5±4.4 Experimental group 1 8.83±0.72* 17.8±6.2* Experimental group 2 10.79±0.82* 20.6±5.6* Experimental group 3 10.67±0.48* 19.7±4.8* Experimental group 4 10.32±0.62* 20.2±6.2*

[0099] Note: * indicates that P < 0.05 compared to the control group.

[0100] As shown in the table above, compared with the blank group, the length and weight of newly grown hair in the experimental group mice were significantly increased (P<0.05). At the same time, the hair growth rate of experimental group 2-4 was higher than that of experimental group 1. Therefore, it can be seen that the combined use of drugs significantly enhances the effect on hair growth.

[0101] 4.3 Number of hair follicles in the growth phase

[0102] At the end of the experiment, skin from the hairless area on the back of the mice was taken, fixed with formalin, and used for pathological sections. After staining, the longitudinal sections and transverse sections at the same level of the mouse hair follicles were observed under a microscope. The results are shown in Table 9.

[0103] Table 9. Number of hair follicles in the growth phase

[0104] Group Number of hair follicles in the growth phase (per field of view) Blank group 37±6 Experimental group 1 54±8* Experimental group 2 69±10* Experimental group 3 71±11* Experimental group 4 70±9*

[0105] Note: * indicates that P < 0.05 compared to the control group.

[0106] As shown in the table above, compared with the control group, the experimental group mice had a significantly higher number of hair follicles in the anagen phase, with the number of hair follicles in phases 2-4 of the experimental group being higher than that in phase 1. This indicates that the combined use of the drugs significantly enhanced the effect on hair growth.

[0107] Example 4: Experiment on the effect of oral administration of cedar oil hair growth tablets on hair growth in mice

[0108] 1. Experimental drugs

[0109] 5% minoxidil topical solution, purchased from Shandong Jingwei Pharmaceutical Co., Ltd., National Drug Approval Number H20233798.

[0110] 2. Laboratory animals

[0111] C57BL / 6 mice, SPF grade, 6-8 weeks old, were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd.

[0112] 3. Experimental Grouping and Experimental Design

[0113] Mice were lightly anesthetized with ether, and their backs were shaved using a rosin / wax mixture in equal proportions, covering an area of ​​approximately 2cm × 3cm. After shaving, the mice were randomly divided into 5 groups of 20 mice each, and the administration regimens are shown in Table 10.

[0114] Table 10. Dosing Regimen

[0115] Group Dosing regimen Blank group Gavage with an equal volume of distilled water experimental group Gavage administration of an equal volume of cedar oil hair growth tablets (prepared as a suspension using distilled water) 40 mg / kg Positive control group Apply 0.1 mL of 5% minoxidil solution evenly.

[0116] After hair removal, administer the medication as described above for 18 consecutive days. Record the hair growth in the bald areas daily, and collect the hair from the bald areas after sacrifice on day 19, measuring and recording the length and weight.

[0117] 4. Experimental Results

[0118] 4.1 Hair growth status

[0119] After hair removal, the skin on the back of the mice turned pink. After the drug was applied, the skin of the experimental group and the positive control group mice began to turn gray on the 5th-6th day after administration. Gray new hairs were visible on the 8th-10th day after administration, and the skin was covered with gray hairs on the 16th-17th day after administration.

[0120] 4.2 Length and weight of new hair growth

[0121] After 19 days, the length and weight of the hair in the bald areas of the mice were measured. The results are shown in Table 11.

[0122] Table 11. Length and weight of newborn mouse fur

[0123] Group New hair growth length (mm) <![CDATA[New hair weight (mg / cm 2 )]]> Blank group 6.73±0.47 14.2±3.1 experimental group 8.72±0.34* 17.3±4.9* Positive control group 9.32±0.39* 18.8±5.1*

[0124] Note: * indicates that P < 0.05 compared to the control group.

[0125] As shown in the table above, compared with the blank group, there were significant differences in the length and weight of newly grown hair in both the experimental group and the positive control group (P<0.05).

[0126] Example 5: Experiment on the effect of cedar oil on the growth of human hair papilla cells

[0127] 1. Experimental Materials

[0128] Human dermal papilla cells were purchased from Qingqi (Shanghai) Biotechnology Development Co., Ltd.; DMEM culture medium and fetal bovine serum were purchased from Gibco, USA.

[0129] 2. Experimental Grouping and Experimental Design

[0130] The drugs were divided into 6 groups and the drug concentrations in the co-culture are shown in Table 12.

[0131] Table 12. Drug Concentration

[0132] Group Drug concentration Blank group DMEM medium Experimental group 1 DMEM medium containing 30 μg / mL cedar oil Experimental group 2 DMEM medium containing 60 μg / mL cedar oil Experimental group 3 DMEM medium containing 160 μg / mL cedar oil Experimental group 4 DMEM medium containing 200 μg / mL cedar oil Positive control group Contains 5% minoxidil

[0133] After co-culturing the above-mentioned drug concentration with human dermal papilla cells, cell proliferation was determined by the MTT assay.

[0134] 3. Experimental Results

[0135] The results are shown in Table 13 below. After co-culture, both the experimental group and the positive control group promoted the proliferation of human dermal papilla cells to varying degrees, compared with the blank group. When the concentration of cedar oil was greater than or equal to 160 μg / mL (i.e., experimental group 3-4), it had a significant effect on the proliferation of dermal papilla cells, and the cell proliferation rate was higher than that of the positive control group, indicating that its activity was higher than that of 5% minoxidil.

[0136] Table 13. Cell proliferation status

[0137] Group Proliferation rate Blank group / Experimental group 1 108.5％ Experimental group 2 114.8％ Experimental group 3 151.4％ Experimental group 4 153.3％ Positive control group 145.8％

[0138] Example 6: Skin irritation test of cedar oil on guinea pigs

[0139] 1. Experimental drugs

[0140] 95% ethanol, purchased from Sinopharm Chemical Reagent Co., Ltd.

[0141] 2. Laboratory animals

[0142] Guinea pigs, SPF grade, weighing 330-380g, purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd.

[0143] 3. Experimental Grouping and Experimental Design

[0144] Six guinea pigs were used in total. After hair was removed from the backs of each guinea pig, they were divided into left and right sections. The administration regimen is shown in Table 14.

[0145] Table 14. Dosing Regimen

[0146] Group Dosing regimen Blank group (left area) 95% ethanol Experimental group (right area) 200mg / kg cedar oil

[0147] After one week of acclimatization, the guinea pigs were dehaired on their backs using a rosin / wax mixture in equal proportions, covering an area of ​​approximately 2cm × 3cm. 95% ethanol was applied to the left side of the guinea pig's back, and 144mg / kg cedar oil was applied to the right side. The treatment was repeated once daily for 7 consecutive days before washing off. The erythema and edema on the skin of the guinea pig's back were then observed visually after the treatment.

[0148] 4. Experimental Results

[0149] Table 15 shows the condition of the guinea pig's back skin. As can be seen from the table, the cedar oil had virtually no irritation to the guinea pig's skin at the four observation points of 1, 24, 48, and 72 hours.

[0150] Table 15. Results of Guinea Pig Skin Allergy

[0151] Group 1h 24h 48h 72h evaluate Blank group Slight redness normal normal normal Non-irritating experimental group Slight redness normal normal normal Non-irritating

[0152] Example 7: Experiment on skin allergy to guinea pigs caused by cedar oil

[0153] 1. Experimental drugs

[0154] 95% ethanol was purchased from Sinopharm Chemical Reagent Co., Ltd.; 1-chloro-2,4-dinitrobenzene (purity: 97%) was purchased from Shanghai Aladdin Biochemical Technology Co., Ltd.

[0155] 2. Laboratory animals

[0156] Guinea pigs, SPF grade, 330-380g, half male and half female, purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd.

[0157] 3. Experimental Grouping and Experimental Design

[0158] Guinea pigs were randomly divided into 3 groups of 5 each, and the dosing regimens are shown in Table 16:

[0159] Table 16. Dosing Regimen

[0160] Group Dosing regimen Blank group 95% ethanol experimental group 200mg / kg cedar oil Positive control group 0.1% 1-chloro-2,4-dinitrobenzene

[0161] After one week of acclimatization, the guinea pigs were sensitized by removing hair from the left side of their backs using a mixture of rosin and wax in equal proportions, covering an area of ​​approximately 3cm x 3cm. The sensitization was then induced by sensitization, and the right side of the guinea pigs was sensitized by stimulation 14 days later.

[0162] Sensitization exposure: On days 1, 7, and 14, the corresponding drugs were applied to the hairless area on the left side of the spine of guinea pigs in the blank group, experimental group, and positive control group to sensitize the guinea pig skin.

[0163] Triggering exposure: 14 days after sensitization, apply the corresponding drug to the hairless area on the right side of the guinea pig. After 6 hours of stimulation, wash it off with warm water. Observe the erythema and edema of the guinea pig's skin at different time points after administration. The skin sensitization of the guinea pig is scored according to the severity (erythema 1-3 points, edema 1-3 points, total 6 points).

[0164] 4. Experimental Results

[0165] Table 17 details the erythema and edema on the back of the guinea pigs. As shown in the table, when observed at 6, 24, 48 and 72 hours, the erythema and edema in the control group and the experimental group were basically 0, that is, no erythema or edema appeared. This indicates that cedar oil does not cause allergic reactions on the skin of guinea pigs.

[0166] Table 17. Results of Guinea Pig Skin Allergy

[0167]

[0168] To facilitate comparative testing, cedrol was added as a control group. To ensure the accuracy and comparability of the experiments, the low, medium, and high dose groups of cedar oil in this invention were replicated. Examples and results are shown below:

[0169] Example 8: Functional experiment of cedar oil promoting hair growth in normal C57BL / 6 mice

[0170] 1. Laboratory animals

[0171] C57BL / 6 mice, SPF grade, 6-8 weeks old, weighing 20±2g, were obtained from Beijing Vital River Laboratory Animal Technology Co., Ltd.

[0172] 2. Animal modeling methods

[0173] After mildly anesthetizing the mice with ether, the backs of the mice were dehaired using a mixture of equal proportions of rosin and paraffin, covering an area of ​​approximately 2cm × 3cm.

[0174] 3. Experimental Grouping and Experimental Design

[0175] A total of 160 mice were randomly divided into 8 groups of 20 mice each. The drug administration regimens are shown in Table 18.

[0176] Table 18. Experimental Groups and Dosing Regimens

[0177]

[0178]

[0179] Apply the medication twice daily, 1 ml each time, to the hair-removed area for 18 consecutive days. Record hair growth in the hair-removed area of ​​the mice. After sacrifice on day 19, collect the hair from the hair-removed area and measure its length and weight. At the end of the experiment, take skin from the hair-removed area on the back of the mice, fix it with formalin, and prepare pathological sections to observe hair follicle growth.

[0180] 4. Experimental Results

[0181] 4.1 Hair growth status

[0182] After hair removal, the skin on the backs of mice turned pink. After drug application, the skin color of mice in the medium- and high-dose cedarwood oil groups darkened and new hair follicles appeared on days 4-5. In the medium- and high-dose cedarol groups, the skin color darkened and new hair follicles appeared on days 5-6. No significant changes were observed in the control group. In the medium- and high-dose cedarwood oil groups, sparse distribution of new gray downy hairs was visible on days 8-10 after administration, and the skin was basically covered by hair by approximately day 16-17. The medium- and high-dose cedarol groups showed a delay of approximately 1-2 days. Compared to the high-dose cedarol group, the high-dose cedarwood oil group showed significantly better hair growth. The positive control group showed little difference from the high-dose cedarol group.

[0183] 4.2 Number of newly formed hair follicles

[0184] After staining, longitudinal sections and transverse sections at the same level of mouse hair follicles were observed under a microscope. The results are shown in Table 19.

[0185] Table 19. Number of newly formed hair follicles

[0186]

[0187]

[0188] Note: * indicates a significant difference compared to the control group, P<0.05; #This indicates a significant difference compared to the high-dose cedrol group (P<0.05).

[0189] As shown in the table above, compared with the blank group, the number of newly formed hair follicles was greater in the cedarwood oil, cedrol, and positive control groups. The number of hair follicles in the medium- and high-dose cedarwood oil and cedrol groups, as well as the positive control group, showed significant differences compared with the blank group (P<0.05). Compared with the high-dose cedrol group, the number of hair follicles in the high-dose cedarwood oil group was significantly increased (P<0.05), indicating that the activity of high-dose cedarwood oil was significantly higher than that of cedrol. Furthermore, the activity of the positive control group was slightly higher than that of the high-dose cedrol group.

[0190] 4.3 Length and weight of new hair growth

[0191] After 19 days, the length and weight of the hair in the bald areas were measured. The results are shown in Table 20.

[0192] Table 20. Length and weight of newborn mouse fur

[0193] Group New hair growth length (mm) <![CDATA[New hair weight (mg / cm 2 )]]> Blank group 6.53±0.38 13.9±2.7 Low-dose group of cedar oil 6.83±0.31 14.5±5.3 Medium dose group of cedar oil 8.23±0.62* 16.5±4.5* High-dose group of cedar oil <![CDATA[9.79±0.56* # ]]> <![CDATA[18.9±6.8* # ]]> low-dose cedrol group 6.68±0.31 14.5±4.5 medium-dose cedrol group 7.89±0.53* 16.0±5.1* High-dose cedrol group 8.89±0.42* 17.1±5.9* Positive control group 9.13±0.28* 17.8±4.1*

[0194] Note: * indicates a significant difference compared to the control group, P<0.05; # This indicates a significant difference compared to the high-dose cedrol group (P<0.05).

[0195] As shown in the table above, compared with the blank group, the length and weight of newly grown hair were significantly increased in the high-dose groups of cedarwood oil and cedrol, as well as the positive control group (P<0.05). Compared with the high-dose cedrol group, the hair growth value of mice in the high-dose cedarwood oil group was significantly higher than that in the cedrol group (P<0.05), while the hair growth value of the high-dose cedrol group was slightly lower than that in the positive control group. This indicates that the activity of high-dose cedarwood oil in promoting hair growth is significantly higher than that of high-dose cedrol, and the activity of the positive control group is slightly higher than that of high-dose cedrol.

[0196] Example 9: Functional experiment of cedarwood oil promoting hair growth in mice with cyclophosphamide-induced alopecia.

[0197] 1. Laboratory animals

[0198] C57BL / 6 mice, SPF grade, 6-8 weeks old, weighing 20±2g, were obtained from Beijing Vital River Laboratory Animal Technology Co., Ltd.

[0199] 2. Animal modeling methods

[0200] After mildly anesthetizing the mice with ether, the backs of the mice were shaved using a mixture of rosin and wax in equal proportions, covering an area of ​​approximately 2cm × 3cm.

[0201] After hair removal, the test drug was applied twice daily. After 7 days of administration, mice were given a single intraperitoneal injection of cyclophosphamide 150 mg / kg. -1If large patches of hair fall out 4-5 days after injection, the animal model has been successfully prepared.

[0202] 3. Experimental Grouping and Experimental Design

[0203] A total of 160 mice were randomly divided into 8 groups of 20 mice each. The drug administration regimens are shown in Table 21.

[0204] Table 21. Experimental Groups and Dosing Regimens

[0205] Group Dosing regimen Blank group Apply an equal amount of 95% ethanol evenly Low-dose group of cedar oil Apply an equal amount of 20 mg / kg of 95% ethanol solution of cedar oil evenly. Medium dose group of cedar oil Apply an equal amount of 95% ethanol solution of cedar oil at a concentration of 40 mg / kg evenly. High-dose group of cedar oil Apply an equal amount of 95% ethanol solution of cedar oil at a concentration of 80 mg / kg evenly. low-dose cedrol group Apply an equal amount of cedrol solution (10 mg / kg) evenly. medium-dose cedrol group Apply an equal amount of cedrol solution (20 mg / kg) evenly. High-dose cedrol group Apply an equal amount of cedrol solution (40 mg / kg) evenly. Positive control group Apply an equal amount of 2% minoxidil lotion evenly.

[0206] After hair removal, apply the medication twice daily, and after 7 days use 150mg / kg. -1 Cyclophosphamide was used to establish a hair loss model in mice, and the mice were administered the drug for 18 days. Hair growth in the bald areas was recorded, and the mice were sacrificed on day 19 and the hair from the bald areas was collected for measurement of length and weight. At the end of the experiment, skin from the bald areas on the backs of the mice was taken, fixed with formalin, and used for pathological sectioning to observe hair follicle growth.

[0207] 4. Experimental Results

[0208] 4.1 Hair growth status

[0209] After hair removal, the skin on the backs of mice turned pink. On days 2-3, the balded area began to turn gray; on days 5-6, it was covered with gray downy hair; and on days 8-9, it was almost completely covered with hair. Approximately 2-3 days after cyclophosphamide injection, mice began to experience hair loss; on days 4-5, large areas of hair fell out, indicating successful model establishment. After 18 days of continuous administration, compared to the control group, hair growth was significantly increased in the high-dose cedrol and cedarwood oil groups and the positive control group. Compared to the high-dose cedrol group, the high-dose cedarwood oil group showed significantly increased hair growth. The high-dose cedrol group was comparable to the positive control group.

[0210] 4.2 Number of newly formed hair follicles

[0211] After staining, longitudinal sections and transverse sections at the same level of mouse hair follicles were observed under a microscope. The results are shown in Table 22.

[0212] Table 22. Number of newly formed hair follicles

[0213] Group Number of newly formed hair follicles (per field of view) Blank group 23±8 Low-dose group of cedar oil 30±10 Medium dose group of cedar oil 47±12* High-dose group of cedar oil <![CDATA[63±14* # ]]> low-dose cedrol group 28±9 medium-dose cedrol group 41±12* High-dose cedrol group 53±14* Positive control group 56±10*

[0214] Note: * indicates a significant difference compared to the control group, P<0.05; # This indicates a significant difference compared to the high-dose cedrol group (P<0.05).

[0215] As shown in the table above, compared with the control group, the number of newly formed hair follicles in the cedarwood oil group, cedarol group, and positive control group was greater. Among them, the number of hair follicles in the high-dose cedarwood oil and cedarol groups and the positive control group were significantly different from the control group (P<0.05). Compared with the high-dose cedarol group, the number of newly formed hair follicles in the high-dose cedarol oil group was significantly higher (P<0.05), while the number of newly formed hair follicles in the positive control group was slightly higher than that in the high-dose cedarol group. This indicates that the activity of high-dose cedarwood oil is significantly higher than that of high-dose cedarol, and the activity of the positive control group is slightly higher than that of high-dose cedarol.

[0216] 4.3 Length and weight of new hair growth

[0217] After 19 days, the length and weight of the hair in the bald areas were measured. The results are shown in Table 23.

[0218] Table 23. Length and weight of newborn mouse fur

[0219] Group New hair growth length (mm) <![CDATA[New hair weight (mg / cm 2 )]]> Blank group 4.86±0.33 12.2±2.1 Low-dose group of cedar oil 6.02±0.37 14.1±3.5 Medium dose group of cedar oil 7.42±0.48* 16.1±4.1* High-dose group of cedar oil <![CDATA[8.39±0.23* # ]]> <![CDATA[17.6±3.4* # ]]> low-dose cedrol group 5.72±0.35 13.8±2.6 medium-dose cedrol group 6.56±0.62* 15.2±4.5* High-dose cedrol group 7.69±0.37* 16.4±4.6* Positive control group 7.88±0.38* 16.7±2.8*

[0220] Note: * indicates a significant difference compared to the control group, P<0.05; # This indicates a significant difference compared to the high-dose cedrol group (P<0.05).

[0221] As shown in the table above, compared with the blank group, there were significant differences in the length and weight of newly grown hair in the high-dose group of Chinese fir oil, the high-dose group of cedrol, and the positive control group (P<0.05). Compared with the cedrol group, the length and weight of newly grown hair in the high-dose group of Chinese fir oil were significantly higher than those in the cedrol group (P<0.05), while the length and weight of newly grown hair in the positive control group were similar to those in the high-dose group of cedrol. This indicates that the activity of high-dose Chinese fir oil is significantly higher than that of high-dose cedrol, while the activity of the positive control group is not significantly different from that of high-dose cedrol.

[0222] Example 10: Experiment on the effect of cedarwood oil in combination with other drugs on hair growth in normal C57BL / 6 mice

[0223] 1. Laboratory animals

[0224] C57BL / 6 mice, SPF grade, 6-8 weeks old, weighing 20±2g, were obtained from Beijing Vital River Laboratory Animal Technology Co., Ltd.

[0225] 2. Animal modeling methods

[0226] After mildly anesthetizing the mice with ether, the backs of the mice were dehaired using a mixture of equal proportions of rosin and paraffin, covering an area of ​​approximately 2cm × 3cm.

[0227] 3. Experimental Grouping and Experimental Design

[0228] A total of 180 mice were randomly divided into 9 groups of 20 mice each. The drug administration regimens are shown in Table 24.

[0229] Table 24. Experimental Groups and Dosing Regimens

[0230]

[0231] The medication was applied twice daily to the hair-removed area for 18 consecutive days. Hair growth in the hair-removed area was recorded, and the mice were sacrificed on day 19. The hair from the hair-removed area was collected and its length and weight were measured. At the end of the experiment, skin from the hair-removed area on the back of the mice was taken, fixed with formalin, and prepared for pathological sectioning to observe hair follicle growth.

[0232] 4. Experimental Results

[0233] 4.1 Hair growth status, length and weight of new hair growth

[0234] Compared to the control group, the experimental group mice had denser and better-colored hair in the bald areas. After sacrifice at 19 days, the length and weight of the hair in the bald areas were measured. The results are shown in Table 25:

[0235] Table 25. Length and weight of newborn mouse fur

[0236] Group New hair growth length (mm) <![CDATA[New hair weight (mg / cm 2 )]]> Blank group 6.98±0.48 15.8±4.4 Experimental group 1 8.32±0.72 17.5±6.2 Experimental group 2 <![CDATA[10.59±0.82* #◇ ]]> <![CDATA[21.0±5.6* #◇ ]]> Experimental group 3 <![CDATA[10.67±0.48* # ]]> <![CDATA[21.4±4.8* # ]]> Experimental group 4 <![CDATA[10.81±0.62* #Δ ]]> <![CDATA[21.7±5.3* #Δ ]]> Experimental group 5 7.67±0.46 15.8±5.2 Experimental group 6 9.62±0.68* 18.3±6.1* Experimental group 7 9.65±0.47* 18.9±5.3* Experimental group 8 9.92±0.72* 19.1±6.2*

[0237] Note: * indicates a significant difference compared to the control group, P<0.05; # This indicates a significant difference compared to experimental group 1, P<0.05; ◇ This indicates a significant difference compared to experimental group 6 (P<0.05); and a significant difference compared to experimental group 7 (P<0.05). Δ This indicates a significant difference compared to experimental group 8, P<0.05.

[0238] As shown in the table above, compared with the control group, the length and weight of newly grown hair in the experimental group increased. Furthermore, the hair growth rate in experimental groups 2-4 was significantly higher than that in experimental group 1 (P<0.05). This indicates that the combined use of the drugs significantly enhanced the effect on hair growth. In addition, pairwise comparisons showed that the hair growth rate in experimental groups 2-4 was significantly higher than that in experimental groups 6-8 (P<0.05), indicating that the combined use of cedarwood oil had significantly higher activity than cedrol.

[0239] 4.2 Number of newly formed hair follicles

[0240] After staining, longitudinal sections and transverse sections at the same level of the mouse hair follicles were observed under a microscope. The results are shown in Table 26.

[0241] Table 26. Number of newly formed hair follicles

[0242]

[0243]

[0244] Note: * indicates a significant difference compared to the control group, P<0.05; # This indicates a significant difference compared to experimental group 1, P<0.05; ◇ This indicates a significant difference compared to experimental group 6 (P<0.05); and a significant difference compared to experimental group 7 (P<0.05). Δ This indicates a significant difference compared to experimental group 8, P<0.05.

[0245] As shown in the table above, compared with the control group, the experimental group had a greater number of newly formed hair follicles, with the number of hair follicles in experimental group 2-4 being significantly higher than that in experimental group 1 (P<0.05). This indicates that the combined use of the drugs significantly enhanced the effect on hair growth. Furthermore, pairwise comparisons showed that the hair growth rate in experimental group 2-4 was higher than that in experimental group 6-8 (P<0.05), indicating that the activity of cedarwood oil was higher than that of cedrol after combined use.

[0246] Example 11: Experiment on the effect of oral administration of cedar oil hair growth tablets on hair growth in mice

[0247] 1. Laboratory animals

[0248] C57BL / 6 mice, SPF grade, 6-8 weeks old, weighing 20±2g, were obtained from Beijing Vital River Laboratory Animal Technology Co., Ltd.

[0249] 2. Animal modeling methods

[0250] After mildly anesthetizing the mice with ether, the backs of the mice were shaved using a mixture of rosin and wax in equal proportions, covering an area of ​​approximately 2cm × 3cm.

[0251] 3. Experimental Grouping and Experimental Design

[0252] Eighty mice were randomly divided into four groups of 20 each, and the administration regimens are shown in Table 27.

[0253] Table 27. Experimental Groups and Dosing Regimens

[0254]

[0255]

[0256] After hair removal, administer the medication as described above for 18 consecutive days. Record the hair growth in the bald areas daily, and collect the hair from the bald areas after sacrifice on day 19, measuring and recording the length and weight.

[0257] 4. Experimental Results

[0258] 4.1 Hair growth status

[0259] After hair removal, the skin on the backs of mice turned pink. After drug application, the skin in the cedarwood oil group and the positive control group began to turn gray on days 5-6 after administration. Gray new hairs were visible on days 8-10 after administration, and the skin was covered with gray hairs on days 16-17 after administration. The activity of the cedarwood oil group was significantly higher than that of the cedrol group; the activity of the cedrol group was slightly lower than that of the positive control group, with no significant difference.

[0260] 4.2 Length and weight of new hair growth

[0261] After 19 days, the length and weight of the hair in the bald areas were measured. The results are shown in Table 28.

[0262] Table 28. Length and weight of newborn mouse fur

[0263] Group New hair growth length (mm) <![CDATA[New hair weight (mg / cm 2 )]]> Blank group 6.59±0.43 13.9±3.7 Chinese fir oil group <![CDATA[9.69±0.35* # ]]> <![CDATA[17.4±4.6* # ]]> Cedar alcohol group 8.81±0.41* 16.4±6.2* Positive control group 9.13±0.34* 16.7±5.1*

[0264] Note: * indicates a significant difference compared to the control group, P<0.05; # This indicates a significant difference compared to the cedrol group (P<0.05).

[0265] As shown in the table above, compared with the blank group, there were significant differences in the length and weight of newly grown hair in the cedarwood oil group, cedarwood alcohol group, and positive control group (P<0.05). Compared with the cedarwood alcohol group, the length and weight of newly grown hair in the cedarwood oil group were significantly higher than those in the cedarwood alcohol group (P<0.05), while the length and weight of newly grown hair in the positive control group were slightly higher than those in the cedarwood alcohol group. This indicates that the activity of cedarwood oil is significantly higher than that of cedarwood alcohol, and the activity of the positive control group is slightly higher than that of cedarwood alcohol.

[0266] Example 12: Experiment on the effect of cedar oil on the growth of human hair papilla cells

[0267] 1. Experimental Grouping and Experimental Design

[0268] The drugs were divided into 6 groups, and the drug concentrations in the co-culture are shown in Table 29.

[0269] Table 29. Experimental Groups and Drug Concentrations

[0270] Group Drug concentration Blank group DMEM medium Experimental group 1 DMEM medium containing 20 mg / kg cedar oil Experimental group 2 DMEM medium containing 40 mg / kg cedar oil Experimental group 3 DMEM medium containing 80 mg / kg cedar oil Positive control group Contains 2% minoxidil

[0271] After co-culturing the above-mentioned drug concentration with human dermal papilla cells, cell proliferation was determined by the MTT assay.

[0272] 2. Experimental Results

[0273] After co-culture, both the experimental group and the positive control group promoted the proliferation of human dermal papilla cells to varying degrees, compared with the blank group. When the concentration of cedarwood oil was greater than or equal to 80 mg / kg, it had a significant effect on the proliferation of dermal papilla cells, and its activity was higher than that of 2% minoxidil.

[0274] Table 30. Cell proliferation status

[0275] Group Proliferation rate Blank group / Experimental group 1 107.7％ Experimental group 2 122.6％ Experimental group 3 156.2％ Positive control group 141.3％

[0276] Example 13: Skin irritation test of cedar oil on guinea pigs

[0277] 1. Laboratory animals

[0278] Guinea pig, SPF grade, weighing 330-380g, purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd.

[0279] 2. Experimental Grouping and Experimental Design

[0280] Six guinea pigs were used in total. After hair was removed from the backs of each guinea pig, they were divided into left and right sections. The administration regimen is shown in Table 31.

[0281] Table 31. Experimental Groups and Dosing Regimens

[0282] Group Dosing regimen Blank group (left area) 95% ethanol Experimental group (right area) 80mg / kg cedar oil

[0283] After one week of acclimatization, the guinea pigs were dehaired using a mixture of rosin and wax in equal proportions. 95% ethanol was applied to the left side of the guinea pig's back, and 80 mg / kg cedar oil was applied to the right side. The application was repeated for 7 days before washing off. The erythema and edema on the skin of the guinea pig's back were observed with the naked eye after the administration.

[0284] 3. Experimental Results

[0285] The condition of the guinea pig's back skin is detailed in Table 32. As can be seen from the table, the cedar oil had virtually no irritation to the guinea pig's skin at the four observation points of 1, 24, 48, and 72 hours.

[0286] Table 32. Results of Guinea Pig Skin Allergies

[0287] Group 1h 24h 48h 72h evaluate Blank group Slight redness normal normal normal Non-irritating experimental group Slight redness normal normal normal Non-irritating

[0288] Example 14: Experiment on skin allergy to guinea pigs caused by cedar oil

[0289] 1. Laboratory animals

[0290] Guinea pigs, SPF grade, 330-380g, half male and half female, purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd.

[0291] 2. Experimental Grouping and Experimental Design

[0292] A total of 15 guinea pigs were randomly divided into 3 groups of 5 each, and the administration regimens are shown in Table 33:

[0293] Table 33. Experimental Groups and Dosing Regimens

[0294]

[0295]

[0296] After one week of acclimatization, the guinea pigs were sensitized by removing hair from the left side of their backs using a mixture of rosin and wax in equal proportions, covering an area of ​​approximately 3cm x 3cm. The sensitization was then induced by sensitization, and the right side of the guinea pigs was sensitized by stimulation 14 days later.

[0297] Sensitization exposure: On days 1, 7, and 14, the corresponding drugs were applied to the hairless area on the left side of the spine of guinea pigs in the blank group, experimental group, and positive control group (see Table 33) to sensitize the guinea pig skin.

[0298] Provocation exposure: 14 days after sensitization, the corresponding drug was applied to the hairless area on the right side of the guinea pig (see Table 33). After 6 hours of provocation, the drug was washed off with warm water. The erythema and edema of the guinea pig's skin were observed at different time points after drug administration. The skin sensitization of the guinea pig was scored according to the severity (erythema 1-3 points, edema 1-3 points, total 6 points).

[0299] 3. Experimental Results

[0300] Table 34 details the erythema and edema on the skin of the guinea pigs. As can be seen from the table, the cedar oil did not cause an allergic reaction to the skin of the guinea pigs at the four observation points of 6, 24, 48 and 72 hours.

[0301] Table 34. Results of Guinea Pig Skin Allergies

[0302]

[0303] The above description, in conjunction with specific preferred embodiments, provides a further detailed explanation of the present invention and should not be construed as limiting the specific implementation of the invention to these descriptions. For those skilled in the art, any equivalent substitutions or obvious modifications made without departing from the inventive concept, resulting in identical performance or application, should be considered to fall within the patent protection scope defined by the submitted claims.

Claims

1. The application of cedar oil in the preparation of drugs that promote hair growth and alleviate hair loss, characterized in that, The cedar oil, by weight percentage, contains the following components: 18-70% α-cedrol, 1-25% α-cedrolene, 0-11% β-cedrolene, 0-5% α-pinene, and 1-5% cypressene.

2. The application according to claim 1, characterized in that, The hair loss mentioned refers to alopecia areata, seborrheic alopecia, chemotherapy-induced alopecia, senile alopecia, or chronic telogen effluvium.

3. The application according to claim 1, characterized in that, The cedar oil is prepared from cedar wood, and the preparation method includes the following steps: The stems, branches, roots, and trunks of Chinese fir are crushed and put into an extraction vessel. Microwave-assisted supercritical CO2 extraction is used to obtain Chinese fir oil.

4. The application according to claim 3, characterized in that, The particle size of the pulverized material is 3-50 mm.

5. The application according to claim 3, characterized in that, The extraction temperature is 30-45℃, the CO2 temperature is 25-75℃, the CO2 flow rate is 1-10L / h, the extraction pressure is 15-55Mpa, and the extraction time is 1-8h.

6. The application according to claim 3, characterized in that, The supercritical CO2 extraction process involves heating with microwaves, with the microwaves intermittently activated and the temperature maintained.

7. The application according to claim 1, characterized in that, The cedar oil can be used alone or in combination with traditional Chinese medicine or chemical drugs that promote hair growth.

8. The application according to claim 7, characterized in that, The medicinal herb in question is ginger.

9. The application according to claim 7, characterized in that, The chemical drug in question is minoxidil.

10. The application according to claim 1, characterized in that, The cedar oil is formulated into topical or oral preparations with a pharmaceutically acceptable carrier.

11. The application according to claim 10, characterized in that, The external preparation is any one of shampoo, hair dye, conditioner, lotion, serum, ointment, gel, patch, tincture or spray; the oral preparation is any one of tablet, capsule, granule, pill or oral liquid.

12. The application according to claim 10, characterized in that, The pharmaceutically acceptable carrier is at least one of the following: filler, binder, disintegrant, lubricant, solubilizer, penetration enhancer, suspending agent, wetting agent, pigment, fragrance, solvent, surfactant, or flavoring agent.

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