4-terpenyl alcohol for use in the preparation of a medicament, cosmetic, or daily-use product for preventing or treating hair loss

Abstract

This invention discloses the application of 4-terpene alcohol, a naturally derived active ingredient, in the preparation of topical medicines, cosmetics, and other daily chemical products with the efficacy of preventing or treating hair loss. This invention discloses that 4-terpene alcohol can be used as the sole active ingredient to significantly promote hair growth in a hair loss model mouse. The efficacy of different optical isomers shows a trend where the levorotatory isomer (-)-4-terpene alcohol is superior to the dextrorotatory isomer (+)-4-terpene alcohol, and is also superior to the racemic mixture (±)-4-terpene alcohol. Furthermore, the toxicity to human dermal papilla cells shows a trend where (-)-4-terpene alcohol < (+)-4-terpene alcohol < (±)-4-terpene alcohol, thus (-)-4-terpene alcohol is preferred. Compared with the model group, the treated mice showed extremely significant improvements in hair weight, coverage, average length, and hair shaft diameter; pathological sections showed an increase in the number of hair follicles, and improvements in the ratio of anagen to telogen phases and the ratio of epidermal to dermal thickness. In this invention, the dosage of 4-terpene alcohol is 0.425% to 5% by mass, with 1.7% showing better efficacy and lower toxicity, demonstrating its potential application in the preparation of topical products for preventing or combating hair loss and promoting hair growth.

Description

Technical Field

[0001] This invention relates to the fields of medicine and cosmetics, specifically to the application of 4-terpene alcohol in the preparation of topical medicines, cosmetics, and daily chemical products with the effect of preventing or treating hair loss. Technical Background

[0002] Hair loss is a skin appendage disease characterized by abnormal hair loss, reduced hair density, and decreased hair follicle function. It has become a widely concerned health and beauty issue globally. Currently, the number of people experiencing hair loss in my country has exceeded 200 million, with over 40% under the age of 30. Androgenetic alopecia (AGA) is the most common type of hair loss, accounting for over 70% of all cases. Its core pathogenesis lies in the abnormally elevated levels of androgens (especially dihydrotestosterone) within the hair follicles, based on a genetic predisposition. These androgens bind to androgen receptors on the surface of dermal papilla cells, leading to a shortened follicle growth phase, progressive miniaturization, and ultimately, loss of regenerative capacity. Furthermore, external factors such as scalp microenvironment imbalance (inflammation, microbial infection, insufficient nutrition), as well as indirect factors such as staying up late, mental stress, and malnutrition, can all induce or worsen hair loss symptoms. The clinical characteristics of AGA show significant population differences; men often present with a receding hairline and thinning hair on the crown, while women typically experience diffuse hair loss on the crown.

[0003] Hair loss not only affects appearance but also easily leads to psychological problems such as low self-esteem and anxiety, which in turn affects employment, social life, and romantic relationships. Currently, treatment methods for AGA (Alopecia Areata) mainly include drug therapy (minoxidil, finasteride), physical therapy, surgical hair transplantation, and topical care products. However, existing technologies have significant drawbacks: minoxidil has a slow onset of action, causes local irritation, and is prone to relapse after discontinuation; long-term use of finasteride may lead to side effects such as sexual dysfunction and is contraindicated in a wide range of populations, including women and minors; physical therapy and surgical hair transplantation are costly and complex; and topical care products often have unclear ingredients, unknown mechanisms of action, and inconsistent effectiveness, with some products containing preservatives and surfactants that may even worsen scalp damage.

[0004] 4-Terpenols are naturally occurring monoterpenol compounds widely found in the volatile oils of plants such as tea tree, clove, and thyme, with the highest content (30%–50%) found in the essential oil of Melaleuca alternifolia (tea tree). This compound exists in various chiral isomers, primarily including the naturally dominant (-)-4-terpenol, the less abundant (+)-4-terpenol, and the commonly found synthetic (±)-4-terpenol. Different chiral configurations exhibit significant differences in biological activity. Modern pharmacological studies have shown that this component possesses broad-spectrum antibacterial, anti-inflammatory, and antioxidant activities. It can inhibit bacterial and fungal growth by disrupting the cell membrane structure of microorganisms, and exert anti-inflammatory effects by downregulating inflammatory mediators. Based on these properties, 4-terpenols are currently used clinically as an adjunct treatment for skin inflammation, acne, athlete's foot, eczema, and oral infections, effectively alleviating related symptoms. Its applications span across pharmaceuticals, daily chemicals, and food and agriculture: in pharmaceuticals, it is mainly used to prepare topical anti-inflammatory and antibacterial agents; in daily chemicals, it is widely added to skincare and personal care products for acne treatment, oil control, dandruff removal, and itch relief; and in food and agriculture, it shows promising potential as a natural fragrance, preservative, and low-toxicity plant-derived pesticide. Despite its mature applications in multiple fields, its application in hair loss prevention and treatment lacks systematic research, and the differences in the anti-hair loss and hair growth efficacy of different optical isomers are rarely reported. Summary of the Invention

[0005] This invention provides a novel application method of 4-terpene alcohol, namely, its application in the preparation of topical medicines, cosmetics, and daily chemical products with the effect of preventing or treating hair loss, wherein the 4-terpene alcohol is selected from one or more of (-)-4-terpene alcohol, (+)-4-terpene alcohol, or (±)-4-terpene alcohol, preferably (-)-4-terpene alcohol or (+)-4-terpene alcohol, and most preferably (-)-4-terpene alcohol.

[0006] As an improvement of the present invention, the mass fraction of 4-terpene alcohol in topical medicines, cosmetics, and daily chemical products is ≥0.425%, preferably 0.425%~5%, and most preferably 1.7%.

[0007] The hair loss type is selected from androgenetic alopecia, alopecia areata, anagen effluvium, telogen effluvium, traction alopecia, scarring alopecia, postpartum alopecia, nutritional metabolic alopecia, and infectious alopecia, with androgenetic alopecia being the preferred type.

[0008] The topical medicines, cosmetics, and daily chemical products with the function of preventing or treating hair loss may be selected from one of the following dosage forms: tincture, gel, ointment, cream, patch, plaster, foam, lotion, film, patch, microneedle, and microcrystal, with gel being preferred.

[0009] Beneficial effects

[0010] This invention discloses the application of 4-terpene alcohol in the preparation of topical pharmaceuticals, cosmetics, or daily chemical products with preventive or therapeutic effects on hair loss. Experimental results show that 4-terpene alcohol can intervene in a mouse model of androgenetic alopecia induced by testosterone propionate through topical application, without the need for large amounts of excipients or complex preparation processes. Compared with the model group, mice treated with 4-terpene alcohol had thicker and shinier hair, and their hair-related indicators and skin histopathological indicators were significantly improved. The efficacy showed a certain concentration dependence, with 1.7% 4-terpene alcohol showing the best therapeutic effect. Animal experiments showed that the hair growth-promoting effect followed the trend of (-)-4-terpene alcohol > (+)-4-terpene alcohol > (±)-4-terpene alcohol, and the toxicity to human dermal papilla cells followed the trend of (-)-4-terpene alcohol < (+)-4-terpene alcohol < (±)-4-terpene alcohol. Its mechanism of action may be related to the compound's antibacterial properties, promoting hair growth by regulating sebum secretion and improving the seborrheic microenvironment of the scalp. Within the effective concentration range, 4-terpene alcohol showed no significant irritation to either intact or broken skin, and no allergic reactions were observed. HE staining of mouse heart, liver, spleen, lung, and kidney tissues showed no significant pathological changes, indicating good safety. Based on this, 4-terpene alcohol has promising development prospects in the fields of cosmetics and pharmaceutical raw materials, and can be used to prepare topical formulations for the prevention or treatment of hair loss. Attached Figure Description

[0011] The present invention will be further described below with reference to the accompanying drawings:

[0012] Figure 1: Preventive effects of different concentrations of (±)-4-terpenoid alcohol on a mouse model of hair loss (n=8).

[0013] Figure 2: Comparison of hair detection indicators in each group of mice (n=8).

[0014] Figure 3: HE-stained sections of mouse skin tissue from each group.

[0015] Figure 4: Effects of hair follicle and dermal indices on skin pathological sections of mice in each group (n=3).

[0016] Figure 5: The therapeutic effects of different optical isomers of 4-terpene alcohol on a mouse model of hair loss (n=8).

[0017] Figure 6: Comparison of hair detection indicators in each group of mice (n=8).

[0018] Figure 7: HE-stained sections of mouse skin tissue from each group.

[0019] Figure 8: Comparison of skin hair follicle and dermal indices in each group of mice (n=3).

[0020] Figure 9: HE staining sections of major organs of mice in each group and safety evaluation.

[0021] Figure 10: Effects of different optical isomers of 4-terpene alcohol on the viability of human dermal papilla cells (n=6).

[0022] For all figures, *P<0.05, **P<0.01, ***P<0.001; except for the model group, all other groups are compared with the model group and the differences are marked as statistically significant. Detailed Implementation

[0023] The following specific embodiments further illustrate the technical solution of the present invention, but are not intended to limit the scope of the invention. Tween 80, Span, polyoxyethylene hydrogenated castor oil, poloxamer, lecithin, sodium lauryl sulfate, benzalkonium chloride, and betaines are all commonly used surfactants in topical preparations. In the following embodiments, the most commonly used Tween 80 is selected for testing, but this is not intended to limit the scope of the invention.

[0024] Example 1: Preparation of blank hydrogel

[0025] Weigh 20 mL of purified water into a clean beaker, slowly add 0.2 g of carbomer, and stir magnetically at high speed for 30 min until fully swollen; then slowly add 0.17 g of Tween 80, and continue to stir magnetically for 5 min until well mixed to obtain a mixture; add 1-2 drops of triethanolamine to adjust the pH to obtain a blank hydrogel.

[0026] Example 2: Preparation of 4-terpene alcohol gel

[0027] Weigh 20 mL of purified water into a clean beaker, slowly add 0.2 g of carbomer, and stir magnetically at high speed for 30 min until fully swollen; then add 0.17 g of Tween 80 and the corresponding amount of 4-terpene alcohol, and continue stirring magnetically for 5 min until evenly dispersed, obtaining a milky white mixture; add 1-2 drops of triethanolamine to adjust the pH, thus obtaining the 4-terpene alcohol hydrogel with the corresponding mass fraction. By adjusting the amount of 4-terpene alcohol added, 4-terpene alcohol hydrogels with mass fractions of 0.425%, 0.85%, 1.7%, and 5.0% were prepared, respectively.

[0028] Example 3: Efficacy study of topical application of different concentrations of (±)-4-terpene alcohol for anti-androgenic alopecia

[0029] Six-week-old male C57BL / 6 mice were randomly divided into four groups: a control group, a model group, and four groups treated with 0.425% (±)-4-terpene alcohol, 0.85% (±)-4-terpene alcohol, 1.7% (±)-4-terpene alcohol, and 5% (±)-4-terpene alcohol, with eight mice in each group. One day before the model was established, the central dorsal region of the mice was prepared. Soybean oil and testosterone propionate injection solution were mixed at a volume ratio of 4:1 to prepare a 5 mg / mL dilution of testosterone propionate. Except for the control group, mice in all other groups were treated with this dilution in the morning to induce the hair loss model, and in the afternoon, they were given the corresponding concentration of (±)-4-terpene alcohol gel. The control and model groups were treated with the blank gel. On days 0, 7, 14, and 21 of the experiment, photographs were taken of the treated areas on the backs of the mice in each group to record hair growth. Results are as follows: Figure 1 As shown, the control group mice had normal hair growth, and their skin was gradually covered with hair. In contrast, hair growth in the model group was significantly inhibited, and the skin on their backs was almost smooth and hairless, indicating a successful model. After 21 days of administration, hair growth in all treatment groups was significantly improved compared to the model group. The 1.7% (±)-4-terpene alcohol group had denser and shinier hair than the other treatment groups. Although the 5% (±)-4-terpene alcohol group showed some hair regeneration, the coverage density and uniformity were lower than those in the 1.7% group, possibly because the high concentration stimulated the skin and inhibited hair follicle growth. The overall ranking of treatment effects was: 1.7% (±)-4-terpene alcohol group > control group ≈ 5% (±)-4-terpene alcohol group ≈ 0.85% (±)-4-terpene alcohol group > 0.425% (±)-4-terpene alcohol group > model group. On day 21 of the experiment, mice in the 5% (±)-4-terpene alcohol group, 1.7% (±)-4-terpene alcohol group, 0.425% (±)-4-terpene alcohol group, blank group, and model group had all newly grown hair removed from their backs, and were placed in pre-weighed sealed plastic bags. The weight of the hair was measured and recorded. Simultaneously, ImageJ software was used to analyze back images before administration (day 0) and after administration (day 21) to calculate hair coverage. In addition, 30 hairs were randomly selected from each group, and the average diameter and length of the hair shaft were measured using ImageJ software. The results are as follows: Figure 2 As shown, the ranking of hair weight, coverage, average hair shaft diameter, and average length in the four groups of mice was consistent: 1.7% (±)-4-terpene alcohol group > blank group ≈ 5% (±)-4-terpene alcohol group > 0.425% (±)-4-terpene alcohol group > model group. Based on these indicators, 1.7% (±)-4-terpene alcohol showed the most significant effect in the treatment and prevention of hair loss.

[0030] Example 4: Hematoxylin and eosin (HE) staining of mouse skin tissue

[0031] Skin samples of 1 cm × 1 cm from the back of mice were fixed with 4% paraformaldehyde, trimmed into 0.5 cm × 0.5 cm pieces, and sequentially dehydrated with graded ethanol, cleared with xylene, impregnated with paraffin, and embedded to prepare 5 μm thick sections, which were then baked in a 60℃ oven. The sections were then dewaxed with xylene (I, II) for 15 min each, hydrated with graded ethanol, stained with hematoxylin for 5 min, rinsed with running water for 15 min for bluing, then dehydrated with 75% and 90% ethanol for 10 min each, stained with 0.5% eosin ethanol for 2 min, and rinsed with running water for 1 min. The stained sections were then dehydrated with anhydrous ethanol and cleared with xylene for 3 min each, repeated once, mounted with neutral resin, and observed under a microscope. Under the same magnification and observation area, three pathological sections were selected from each group, and three fields of view were randomly selected from each section. ImageJ was used for statistical analysis, and the detection indicators included the number of hair follicles, the ratio of hair follicle anagen phase to telogen phase, and the ratio of epidermal to dermal thickness. The results are as follows: Figure 3 and Figure 4 As shown.

[0032] In the control group, the hair follicle structure was intact, the hair bulb was enlarged and located deep in the dermis, the dermis and subcutaneous fat layer around the hair follicle were thicker, the hair follicle density was high, and the vast majority of hair follicles were in the anagen phase with clearly visible hair shafts. In the model group, the skin of mice was significantly thinner, the subcutaneous fat layer atrophied, the number of hair follicles was significantly reduced, the structure was disordered, a large number of hair follicles were in the telogen phase, and their volume was reduced, exhibiting typical hair follicle miniaturization changes; the epidermal / dermal thickness ratio was significantly increased. Compared with the model group, all treatment groups showed varying degrees of improvement. In the 0.425% (±)-4-terpene alcohol group, the number of hair follicles increased, some hair follicles entered the anagen phase, the hair bulb enlarged, and the thickness of the skin and dermis recovered somewhat, but the hair follicle size remained uneven. The skin tissue structure in the 1.7% (±)-4-terpene alcohol group was closer to normal, with a significant increase in hair follicle density, returning to levels close to the control group. Numerous hair follicles were in the anagen phase and had regular morphology, with full hair bulbs extending deep into the subcutaneous fat layer. The epidermal / dermal thickness ratio returned to the normal range. The skin tissue structure in the 5% (±)-4-terpene alcohol group also recovered well, but the hair follicle density was lower than that in the 1.7% group. Statistical analysis showed that compared with the model group, the number of hair follicles was significantly increased in the control group, the 1.7% and 5% (±)-4-terpene alcohol groups (p<0.001), and slightly increased in the 0.425% (±)-4-terpene alcohol group. The anagen / telogen ratio of hair follicles was significantly increased in all treatment groups (p<0.001). The epidermal / dermal thickness ratio in the model group was significantly higher than that in the control group and all treatment groups (p<0.001), and this ratio returned to levels close to the control group after treatment.

[0033] Example 5: Pharmacological study of topical application of 4-terpene alcohols with different optical isomers in the treatment of androgenetic alopecia.

[0034] The model was established according to the method in Example 3, and the optimal drug concentration of 1.7% was used for administration. Mice were divided into four groups: blank group, model group, (-)-4-terpene alcohol group, (+)-4-terpene alcohol group, and (±)-4-terpene alcohol group (n=8 in each group). The ameliorative effect of different optical isomers on androgenetic alopecia model was investigated. Photographs of the treated areas on the backs of mice in each group were taken on days 0, 7, 14, 21, and 28 of the experiment to record hair growth. The results are as follows: Figure 5 As shown, all treatment groups promoted hair growth, with the 1.7% (-)-4-terpene alcohol group showing the best effect, close to the level of the blank group, followed by the 1.7% (+)-4-terpene alcohol group and the 1.7% (±)-4-terpene alcohol group. The model group had the slowest hair growth.

[0035] After sampling, the hair weight, hair coverage, hair shaft length, and hair shaft diameter of each group of mice were measured. The results are as follows: Figure 6 As shown in the figure, the hair weight, coverage, and hair shaft diameter in the model group were significantly lower than those in the control group, indicating successful modeling. Compared with the model group, the three optical isomers showed significant differences in promoting hair growth and improving hair follicle growth status, with the efficacy in the order of (-)-4-terpenol > (+)-4-terpenol > (±)-4-terpenol. This suggests that the optical configuration has a significant impact on the efficacy of this type of compound in preventing androgenetic alopecia, and (-)-4-terpenol is preferred as the active ingredient.

[0036] Example 6: Hematoxylin and eosin (HE) staining of mouse skin and major organs

[0037] Following the method in Example 4, HE staining was performed on mouse skin tissues from different optical isomer groups. The number of hair follicles, the ratio of anagen to telogen phases, and the ratio of epidermal thickness to dermal thickness were statistically analyzed. Results are as follows: Figure 7 and Figure 8 As shown, the model group exhibited a significant decrease in the number of hair follicles, a reduced ratio of anagen to telogen follicles, and an increased ratio of epidermal to dermal thickness, displaying typical hair follicle miniaturization and dermal atrophy. Compared to the model group, the pathological morphology of all treatment groups showed improvement. The 1.7% (-)-4-terpene alcohol group recovered to near-blank levels in terms of hair follicle number and anagen phase ratio, with the most intact tissue structure; the 1.7% (+)-4-terpene alcohol group was second best; the 1.7% (±)-4-terpene alcohol group also showed an increase in hair follicle number and anagen follicles, but the improvement was relatively weaker. Three mice were randomly selected from each group, and their heart, liver, spleen, lung, and kidney tissues were subjected to HE staining. Figure 9 As shown, no obvious pathological changes, inflammatory cell infiltration, or tissue necrosis were observed in the major organs and tissues of mice in both the drug-treated groups and the blank group, indicating that 1.7% 4-terpene alcohol had no obvious toxic side effects on the major organs of mice and had good safety.

[0038] Example 7: Effect of 4-terpene alcohols on the viability of human dermal papilla cells

[0039] (-)-4-terpenol, (+)-4-terpenol and (±)-4-terpenol were prepared into stock solutions using dimethyl sulfoxide (DMSO) as solvent. These stock solutions were then diluted with complete culture medium to 0.1%, 0.075%, 0.05%, 0.025% and 0.005% (v / v) working solutions, respectively. The final concentration of DMSO in each group did not exceed 0.1%.

[0040] Logarithmic growth phase human dermal papilla cells (hDPCs) were digested and seeded at 5 × 10³ cells / well in 96-well plates. After 24 h of culture, the cells adhered, and 100 μL of medium containing three different concentrations of 4-terpene alcohols was added to each well, with 6 replicates per group. A normal cell control group (NC) and a blank control group (cell-free) were also included. After another 24 h of culture, 10 μL of LCK-8 solution was added to each well, and the cells were incubated for 2 h. The absorbance at 450 nm was measured using a microplate reader (reference wavelength 630 nm). Cell viability was calculated using the following formula:

[0041] Cell viability (%) = [(OD) 给药组 -OD 空白组 ) / (OD 正常细胞对照组 - OD 空白组 )] × 100%

[0042] The results are as follows Figure 10 As shown in the figure, compared with the normal cell control group, the three optical isomers had no significant effect on hDPC activity at a concentration of 0.005% (v / v). With increasing concentration, (-)-4-terpenol inhibited cell viability at concentrations of 0.05% and above (P < 0.01), while (+)-4-terpenol and (±)-4-terpenol showed significant inhibitory effects at concentrations of 0.025% and above (P < 0.001). The cytotoxicity of the three isomers showed a trend of (±)-4-terpenol > (+)-4-terpenol > (-)-4-terpenol.

[0043] Example 8: Preparation of topical formulations of 4-terpene alcohols for the treatment or prevention of hair loss

[0044] 4-Terpenicillin alcohol is used as a raw material in the preparation of topical formulations, which include topical medicines, cosmetics, and daily chemical products, including the following dosage forms: tinctures, gels, ointments, creams, patches, plasters, foams, lotions, films, patches, microneedles, and microcrystals, with gels being the most preferred. The hair loss described includes androgenetic alopecia, alopecia areata, anagen telogen effluvium, telogen effluvium, traction alopecia, cicatricial alopecia, postpartum alopecia, nutritional metabolic alopecia, and infectious alopecia, with androgenetic alopecia being the most preferred.

Claims

The application of 1,4-terpene alcohols in the preparation of topical medicines, cosmetics, and daily chemical products with the effect of preventing or treating hair loss is characterized by: The 4-terpene alcohol is selected from one or more of (-)-4-terpene alcohol, (+)-4-terpene alcohol, or (±)-4-terpene alcohol.

2. The application as described in claim 1, characterized in that: The 4-terpene alcohol is selected from (-)-4-terpene alcohol and (+)-4-terpene alcohol.

3. The application as described in claim 2, characterized in that: The 4-terpene alcohol is (-)-4-terpene alcohol.

4. The application as described in claim 1 or 3, characterized in that: The 4-terpene alcohol has a mass fraction of 0.425% to 5% in topical medicines, cosmetics, and daily chemical products.

5. The application as described in claim 4, characterized in that: The 4-terpene alcohol has a mass fraction of 0.425% to 1.7% in topical medicines, cosmetics, and daily chemical products.

6. The application as described in claim 5, characterized in that: The mass fraction of the 4-terpene alcohol in topical medicines, cosmetics, and daily chemical products is 1.7%.

7. The application as described in any one of claims 1 to 6, characterized in that: The hair loss mentioned includes androgenetic alopecia, alopecia areata, anagen baldness, telogen effluvium, traction alopecia, scarring alopecia, postpartum alopecia, nutritional metabolic alopecia, and infectious alopecia.

8. The application as described in claim 7, characterized in that: The hair loss described is androgenetic alopecia.

9. The application as described in any one of claims 1 to 8, characterized in that: The dosage forms of the external medicines, cosmetics, and daily chemical products include tinctures, gels, ointments, creams, patches, plasters, foams, lotions, films, patches, microneedles, and microcrystals.

10. The application as described in claim 9, characterized in that: The dosage form of the external medicines, cosmetics, and daily chemical products mentioned is a gel.

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