CN114306192A

CN114306192A - Application of cannabidiol and combination of cannabidiol and cubilose extract in skin moisturizing

Info

Publication number: CN114306192A
Application number: CN202011076296.7A
Authority: CN
Inventors: 詹华强; 白磐竹; 吴启韵; 董婷霞
Original assignee: Hong Kong University of Science and Technology HKUST
Current assignee: Hong Kong University of Science and Technology HKUST
Priority date: 2020-10-10
Filing date: 2020-10-10
Publication date: 2022-04-12
Also published as: WO2022073437A1

Abstract

The present invention relates to the moisturizing efficacy of Cannabidiol (CBD) itself and compositions comprising Cannabidiol (CBD) and cubilose extract on skin cell tissue. Cannabidiol (CBD) and the composition can enhance intercellular water retention capacity and moisture of skin tissues by improving transcription levels of Aquaporin 3(AQP3) gene and Filaggrinn-2 (FLG2) gene of skin cell outer glycoprotein in water channel of skin cells, so that the skin can achieve effects of deep water supplement and skin luster recovery, has no proliferation effect on skin stratum corneum cells, has no obvious stimulation effect on the skin, and is a safe and efficient water supplement humectant.

Description

Application of cannabidiol and combination of cannabidiol and cubilose extract in skin moisturizing

Technical Field

The invention relates to moisturizing of skin; more particularly, the present invention relates to the use of cannabidiol and its combination with cubilose extract for skin hydration.

Background

Modern people have fast pace of life, high pressure and poor sleep, and in addition, environmental pollution is increasingly serious, ultraviolet rays, computer and electric appliance radiation, smoking and drinking, insufficient exercise, excessive consumption of processed foods and excessive intake of oil and fat and the like cause various skin problems, such as dry skin, desquamation, large pores, acne, darkness, uneven skin color, loose skin and the like. It is well known that almost every skin problem is associated with moisture retention and replenishment. Moisture is the first factor of beautiful skin, and whitening, sun protection, oil control and the like are completed on the basis of moisture preservation and water supplement. However, water deficiency has become the origin of skin aging of contemporary women, when the skin is lack of water, cell metabolism, information transmission and the like are affected, the physiological structure of the skin changes such as dry skin, roughness, loss of luster, elasticity reduction, wrinkle increase or deepening, pigment increase, and at worst, the skin barrier dysfunction caused by water deficiency increases the susceptibility of skin diseases and possibly induces and aggravates skin diseases such as dermatitis, eczema, psoriasis, acne and the like.

Most of the existing topical skin preparations increase the hydration level of the skin by reducing evaporation, and generally include naturally occurring skin lipids, sterols, and artificial or natural oils. However, problems associated with such preparations are insufficient moisturizing ability, heavy and greasy feeling, irritation to the skin, and the like.

Therefore, there is an urgent need in the art for a solution that can safely and efficiently moisturize and moisturize skin cells.

Disclosure of Invention

As described above, there is a need in the art for a solution that can safely and efficiently replenish moisture to the skin and maintain the moisture of the skin, thereby achieving a moisturized and whitened skin. Thus, the present invention provides a solution to the problems associated with dry skin and water deficit.

The inventor of the application unexpectedly finds that Cannabidiol (CBD) can improve the gene expression of Aquaporin 3(AQP3) and skin outer layer glycoprotein Filaggrinrin-2 (FLG2) in skin cells, thereby realizing the water replenishing and moisturizing of the skin cells. When Cannabidiol (CBD) and cubilose extract are further combined for use, the two can synergistically improve the expression of AQP3 and FLG2 coding genes, so that the hydration degree of skin is improved, the skin wettability is increased, the intercellular water retention capacity is enhanced, the skin is conditioned and moistened, and the effects of moisturizing, whitening and the like are achieved. Based on this finding, the present inventors have completed the present invention.

In a first aspect, the present invention provides the use of cannabidiol for achieving skin cell hydration and moisturization by increasing gene expression of Aquaporin 3(AQP3) and/or skin outer glycoprotein Filaggrin-2(FLG2) in skin cells.

In a second aspect, the present invention provides a composition for moisturizing skin cells comprising cannabidiol and an extract of cubilose.

In a third aspect, the present invention provides a method of increasing gene expression of Aquaporin 3(AQP3) and/or the skin outer glycoprotein filggrin-2 (FLG2) in a skin cell comprising administering cannabidiol and/or a bird's nest extract to said skin cell.

In a fourth aspect, the present invention provides a method of treating skin comprising topically applying the method of the second aspect to skin in need thereof.

The invention has the advantages that: in the invention, cannabidiol or the combination of cannabidiol and cubilose extract is used for treating dry and water-deficient skin, the expression of AQP3 and FLG2 encoding genes can be improved independently or synergistically, so that the problems related to dry and water-deficient skin are solved, and the water-supplementing and moisturizing effects of the skin are realized; in addition, in the invention, the cannabidiol or the combination of the cannabidiol and the cubilose extract can also achieve the functions of whitening, efficiently moisturizing and preserving the skin by improving the hydration degree of the skin, and is safe and effective to the skin without obvious irritation.

Drawings

The accompanying drawings, which are incorporated in and constitute a part of this specification, provide a further explanation of the invention. It is to be understood that the exemplary drawings in the following description are directed to only some embodiments of the invention and are not to be construed as limiting the invention. Other embodiments can be derived from these figures without inventive effort for a person skilled in the art.

FIG. 1 shows a graph of cytotoxicity analysis of keratinocytes HaCaT cells with Cannabidiol (CBD) at different concentrations and with 3mg/mL of cubilose extract.

FIG. 2 shows a graph of cytotoxicity analysis of HaCaT cells with different concentrations of CBD in combination with 3mg/mL of cubilose extract.

FIG. 3 shows a graph of RT-PCR results of FLG2 gene expression after treatment of HaCaT cells with different concentrations of CBD and 3mg/mL of cubilose extract alone.

FIG. 4 is a graph showing the RT-PCR results of AQP3 gene expression after treatment of HaCaT cells with different concentrations of CBD and 3mg/mL of cubilose extract alone.

FIG. 5 shows a graph of RT-PCR results of FLG2 gene expression after treatment of HaCaT cells with a combination of CBD at various concentrations and 3mg/mL of bird's nest extract.

FIG. 6 is a graph showing the RT-PCR results of AQP3 gene expression after treatment of HaCaT cells with a combination of different concentrations of CBD and 3mg/mL of bird's nest extract.

Fig. 7 shows a graph of the results of human skin sensitivity tests performed on 12 volunteers using a cream containing CBD and bird's nest extract, wherein the blue line part is the application area of the cream.

Detailed Description

The present invention will be described in detail below with reference to examples and the accompanying drawings. It is to be understood that the following description is intended to illustrate the present invention by way of example only and is not intended to limit the scope of the invention, which is defined by the appended claims. Also, it is understood by those skilled in the art that modifications may be made to the technical aspects of the present invention without departing from the spirit and gist of the present invention. Unless otherwise specified, the technical means used in the examples are conventional means well known to those skilled in the art.

As described above, the present inventors have unexpectedly found that cannabidiol and bird's nest extract, whether used alone or in combination, can increase the gene expression of Aquaporin 3(AQP3) and dermal outer glycoprotein Filaggrin-2(FLG2), both of which have extremely important effects on controlling water balance inside and outside skin cells, but the effects are not as significant when used alone as when used in combination. Therefore, the cannabidiol and/or the cubilose extract can be used for solving various skin symptoms caused by dry skin and water shortage, such as darkness, roughness, fine lines, loss of luster and elasticity, pigmentation and the like, and further realizing remarkable whitening, water supplementing and moisturizing effects. In addition, the cannabidiol and the cubilose extract have no proliferation effect on skin stratum corneum cells and have no obvious stimulation effect on skin, so that the cannabidiol and the cubilose extract can be used as a local skin preparation to be externally applied to the skin to achieve the purposes of enhancing intercellular water retention capacity, improving the moisture of skin tissues and recovering skin luster.

AQP3 is a subtype of aquaporin (also called aquaporin) most expressed in human skin, is located on cell membranes, is a transporter capable of transporting small molecular substances such as water, glycerol and urea across membranes, mediates the transport of small molecular substances such as water, glycerol and urea across membranes in organisms by forming "pores" on cell membranes, and regulates water balance. AQP3 has been found to be involved in skin hydration, skin barrier function and wound healing, maintaining normal morphology and function of the skin, and is associated with proliferation and migration of keratinocytes and even early differentiation, the pathogenesis of a variety of skin disorders such as atopic dermatitis, vitiligo, skin tumors, psoriasis and the like. It has also been found that down-regulation of AQP3 expression causes natural aging of the skin at non-exposed sites.

The FLG2 protein is one of S100 fusion type protein family members, and is positioned in an epidermal differentiation complex. FLG2 mRNA and protein are mainly present in the keratinized epithelium, particularly in the skin. The expression pattern of FLG2 is limited to the granular upper and horny layers, and is similar to the expression pattern of filaggrin, and is also closely related structurally to filaggrin. The FLG2 protein is involved in epithelial homeostasis, and the encoded protein is essential for skin keratinization, and defects in the gene are closely related to the development of skin diseases. In addition, the FLG2 protein also has the function of protecting the skin barrier.

When cannabidiol is applied to dry and water-deficient skin, the cannabidiol is found to be capable of remarkably improving the expression of AQP3 and FLG2 coding genes, solving various skin symptoms caused by the dry and water-deficient skin, such as darkness, roughness, fine lines, loss of luster and elasticity, pigment deposition and the like, and further realizing remarkable whitening, water supplementing and moisturizing effects. And when the combination of cannabidiol and cubilose extract is applied to dry and water-deficient skin, the combination is found to synergistically improve the expression of AQP3 and FLG2 coding genes, realize the water retention function of skin cells, and further realize remarkable whitening, water supplementing and moisture retention effects. In addition, the cannabidiol and the combination of the cannabidiol and the cubilose extract have no proliferation effect on skin stratum corneum cells and no obvious stimulation effect on skin, so that the cannabidiol can be used as a local skin preparation to be externally applied to the skin to achieve the purposes of enhancing intercellular water retention capacity, improving the moisture of skin tissues and recovering skin luster.

Therefore, the invention provides the technical concept that the cannabidiol and the combination of the cannabidiol and the cubilose extract are used for moisturizing skin cells, and the technical concept is based on the discovery that the cannabidiol and the cubilose extract can improve the gene expression of Aquaporin 3(AQP3) and outer-layer glycoprotein Filaggrinrin-2 (FLG2) in the skin cells, so that the skin can be effectively moisturized and moisturized, and the skin moisturizing composition is safe and has no side effect. The technical concept of the present invention is different from the technical concept of the prior art topical skin moisturizing formulations that moisturize the skin by increasing the hydration level of the skin by reducing evaporation.

Cannabidiol (CBD) is the major active ingredient in industrial cannabis sativa (hemp) in high content, neither addictive nor neurotoxic. In recent years, cannabidiol has been found to be a powerful antioxidant and has a series of physiological activities such as breast cancer metastasis blocking, epilepsy resistance, spasm resistance, rheumatoid arthritis resistance, anxiety resistance, insomnia resistance and the like. Because of strong antioxidant activity, antibacterial and anti-inflammatory effects, acne removing and soothing effects, aging delaying and damaged cell repairing effects, the skin care product has been applied to skin care products to play the effects of removing acne and diminishing inflammation, and is suitable for treating and preventing acne. Furthermore, as demonstrated in the examples below, cannabidiol can increase the expression of AQP3 and FLG2 encoding genes, thereby acting to moisturize and moisturize skin cells.

"moisturizing" refers to providing moisture to skin cells. "moisturizing" generally refers to the ability to allow moisture to enter the deep cells of the skin and to lock the water firmly at the bottom of the skin without losing it. Herein, the term "moisturization" may be used interchangeably with the terms "water retention" and "water lock".

In a specific embodiment, the effective concentration of cannabidiol is from 0.0001% to 0.001% by weight.

In a preferred embodiment, the effective concentration of cannabidiol is 0.0005 wt%.

In yet another specific embodiment, the cannabidiol is further combined with a cubilose extract for skin cell moisturizing.

The bird's nest is a well-known natural nutritional food for moistening the skin, has the effects of moistening lung dryness, nourishing kidney yin and tonifying deficiency, has super-strong oxidation resistance, and can whiten the skin and lighten color spots. The bird's nest extract used in the present invention may be either commercially available or self-extracted.

In one embodiment, the cubilose extract of the invention can be prepared by the following steps: cooking nidus Collocaliae in water for 1-3 hr, preferably for 2 hr, and freeze drying the cooked product.

In an optional embodiment, the bird's nest may be ground and/or soaked prior to being stewed in water.

In yet another specific embodiment, the effective concentration of the cubilose extract is at least 0.01% by weight.

In a preferred embodiment, the effective concentration of the cubilose extract is at least 0.3 wt.%.

In yet another embodiment, the combination of cannabidiol and bird's nest extract moisturizes and moisturizes the skin cells by increasing the hydration level of the skin, increasing the skin moisturization, and/or conditioning and moisturizing the skin.

In a second aspect, the present invention provides a composition for moisturizing skin cells comprising cannabidiol and an extract of cubilose. The composition can be used for moisturizing skin by synergistically increasing gene expression of Aquaporin 3(AQP3) and/or skin outer layer glycoprotein Filaggrin-2(FLG2) in skin cells. The composition can synergistically improve the transcription levels of AQP3 and FLG2 coding genes, so that the water balance inside and outside skin cells is realized, the hydration degree of skin is further remarkably improved, the skin wettability is increased, and/or the skin is conditioned and moistened, and the skin achieves better whitening, moisturizing and moisturizing effects.

In a particular embodiment, the cannabidiol is from 0.0001% to 0.001% by weight of the composition.

In a preferred embodiment, the cannabidiol is 0.0005% by weight of the composition.

It is understood that the composition of the present invention may include the extract of bird's nest. In one embodiment, the cubilose extract is at least 0.01% by weight of the composition.

In a preferred embodiment, the cubilose extract is at least 0.3% by weight of the composition.

In one embodiment, the composition may contain only cannabidiol, cubilose extract and water. In this case, the cannabidiol may be from 0.0001% to 0.001% by weight of the composition, preferably 0.0005% by weight of the composition; the cubilose extract may be at least 0.01% by weight of the composition, preferably at least 0.3% by weight of the composition; the balance being water.

In a preferred embodiment, the cannabidiol may be 0.0005 wt% of the composition; the cubilose extract may be at least 0.3% by weight of the composition; the balance being water.

Of course, it will be understood by those skilled in the art that the composition of the present invention may contain other ingredients, in addition to cannabidiol, bird's nest extract and water, as desired, such as dosage form (liquid, ointment, cream, etc.), use (medical or cosmetic, etc.), mode of application (spray, spread, etc.), and the like.

It is to be understood that in this context the expression "cannabidiol and/or cubilose extract" includes cannabidiol, cubilose extract and both cannabidiol and cubilose extract. The expression "Aquaporin 3(AQP3) and/or the skin outer glycoprotein Filaggrin-2(FLG 2)" has a similar meaning.

In a specific embodiment, the effective concentration of the cubilose extract is at least 0.01% by weight.

In a fourth aspect, the present invention provides a method of treating skin comprising topically applying to skin in need thereof a composition according to the second aspect of the present invention.

In the context of the present invention, "topical application" means applying the composition of the present invention by painting, spraying, applying, etc., onto the surface of the skin in need thereof (e.g., the skin of the face, neck, hands, arms, legs, or feet).

In a particular embodiment, the composition is applied to skin in need of moisturizing.

In a further specific embodiment, the composition is applied to skin that has fine lines, dullness, uneven skin tone, dryness, peeling, laxity, and/or inelasticity due to water deficit, thereby solving various skin problems caused by water deficit.

The following examples will further illustrate the technical solutions and benefits of the present invention to those skilled in the art, but are not intended to limit the present invention.

Examples

Example 1: compounding nidus Collocaliae extract with Cannabidiol (CBD)

The active ingredients of the present invention are formulated in the following manner:

stock solution of bird's nest extract: 3g of bird's nest (purchased from Hongkong, Hongkong) was weighed, added with 300mL of water, heated and cooked for 2 hours, and then freeze-dried to obtain a bird's nest extract. To 3g of the bird's nest extract, 100mL of water was added to redissolve to prepare a stock solution of bird's nest extract at a concentration of 30 mg/mL.

Cannabidiol (CBD, available from yunnan hansu biotechnology limited) stock solution: cannabidiol 10mg was weighed, a small amount of DMSO was added, stirred until cannabidiol was completely dissolved, and then water was supplemented to 100mL to prepare a cannabidiol stock solution with a concentration of 100 μ g/mL.

Example 2: cytotoxicity assays

To assess whether the compositions of the invention are cytotoxic in keratinocytes and stimulate skin cell proliferation, MTT assays were used and cell viability was analyzed. HaCaT glial cells were cultured in DMEM medium supplemented with 10% FBS (fetal bovine serum), 1% penicillin/streptomycin, respectively. Then, the cultured adherent cells at the bottom of the culture dish were digested with a 0.25% trypsin solution to prepare a cell suspension, which was counted. Then is made intoThe cell concentration was 1X 10⁴one/mL of the suspension was inoculated in a 96-well plate and placed in CO₂Incubator (5% CO)₂Incubated at 37 ℃ for 24 hours. Then adding appropriate amount of cannabidiol stock solution and cubilose extract stock solution respectively to obtain cannabidiol, cubilose extract and combination of cannabidiol and cubilose extract with required concentration. After further culturing for 24 hours, the medium in the 96-well plate was aspirated, and 0.5mg/mL of MTT solution was added and placed in the incubator for 4 hours. After incubation, the cells were analyzed for viability by adding DMSO solution and shaking on a shaker for 15 minutes and measuring the absorbance at 570 nm. The results are shown in FIGS. 1 and 2.

FIG. 1 shows cell viability of HaCaT cells treated separately with cannabidiol at different concentrations (1. mu.g/mL, 2.5. mu.g/mL, 5. mu.g/mL, 10. mu.g/mL, and 25. mu.g/mL), and cubilose extract (3mg/mL), respectively. As compared with the control results, the survival rate of HaCaT cells treated by cannabidiol at a concentration of 1. mu.g/mL to 5. mu.g/mL was improved. Whereas the survival of HaCaT cells was comparable to the control when cannabidiol was present at a concentration of 10 μ g/mL in the composition, but was significantly reduced compared to the control when cannabidiol was present at a concentration of 25 μ g/mL in the composition, showing significant cytotoxicity (p < 0.05).

FIG. 2 shows cell viability of HaCaT cells treated with different concentrations (1. mu.g/mL, 2.5. mu.g/mL, 5. mu.g/mL, 10. mu.g/mL, and 25. mu.g/mL) of cannabidiol in combination with cubilose extract (3mg/mL), respectively. Similarly, as compared with the control results, it was found that when the concentration of cannabidiol was not more than 5 μ g/mL, the survival rate of HaCaT cells was increased, and no cytotoxicity was shown. Cell viability was comparable to controls when cannabidiol was 10 μ g/mL, but HaCaT cell viability was significantly reduced (p <0.01) when cannabidiol was 25 μ g/mL in the composition.

The test results of fig. 1 and 2 show that when the concentration of cannabidiol in the combination of the invention does not exceed 10 μ g/ml, no adverse effect is induced in skin cells and the safety is reliable.

In addition, the present invention also found that neither cannabidiol nor cubilose extract, used alone or in combination, had an effect on HaCaT cell proliferation seen compared to the control (p >0.05, see fig. 1 and 2).

Example 3: effect of cannabidiol and/or cubilose extract on gene expression related to skin moisturizing

To further confirm the moisturizing and moisturizing efficacy of cannabidiol and/or cubilose extracts on skin, the transcription levels of AQP3 and FLG2 encoding genes of skin cells were measured using RT-PCR.

First, HaCaT cells were prepared to a cell concentration of 5X 10⁵one/mL of suspension and seeded on 6-well plates. Put in CO₂Incubator (5% CO)₂Incubated at 37 ℃ for 24 hours. Then adding appropriate amount of cannabidiol stock solution and cubilose extract stock solution respectively to obtain cannabidiol, cubilose extract and combination of cannabidiol and cubilose extract with required concentration. After further incubation for 24 hours, mRNA was extracted from HaCaT cells and RT-PCR was performed using primers for AQP3 and FLG2 encoding genes. The primer sequences are as follows:

AQP 3F-primer: 5'-AGACAGCCCCTTCAGGATTT-3'

AQP 3R-primer: 5'-TCCCTTGCCCTGAATATCTG-3'

FLG 2F-primer: 5'-CTGTGGTCATTCATGGAGTGG-3'

FLG 2R-primer: 5'-CCCTAGAAGGGCTAATGTGTGA-3'

FIG. 3 shows the results of expression of the gene encoding FLG2 obtained by treating HaCaT cells with cannabidiol and bird's nest extract (3mg/mL) alone at different concentrations (1. mu.g/mL, 2.5. mu.g/mL and 5. mu.g/mL), respectively. FIG. 4 shows the results of expression of the genes encoding AQP3 obtained by treating HaCaT cells with cannabidiol and bird's nest extract (3mg/mL) alone at different concentrations (1. mu.g/mL, 2.5. mu.g/mL and 5. mu.g/mL), respectively. As shown in fig. 3 and 4, treatment of skin cells with cannabidiol alone increased gene expression of FLG2 and AQP3 by about 20% to 40%. Particularly, when the concentration of cannabidiol is 2.5 mug/ml, the gene expression effect on FLG2 is best; the best effect on AQP3 gene expression was obtained when cannabidiol concentration was 5. mu.g/ml. HaCaT cells treated with cubilose extract alone showed approximately 10% to 20% increase in transcription levels of FLG2 and AQP3 encoding genes.

These results show that: the cannabidiol or the cubilose extract is used for treating the skin independently, so that the effects of moisturizing and preserving moisture are achieved.

FIG. 5 shows the results of expression of the gene encoding FLG2 obtained by treating HaCaT cells with different concentrations (1. mu.g/mL, 2.5. mu.g/mL and 5. mu.g/mL) of cannabidiol in combination with cubilose extract (3 mg/mL); FIG. 6 shows the results of expression of the genes encoding AQP3 obtained by treating HaCaT cells with different concentrations (1. mu.g/mL, 2.5. mu.g/mL, and 5. mu.g/mL) of cannabidiol in combination with cubilose extract (3 mg/mL). As shown in fig. 5 and 6, HaCaT cells treated with different concentrations of cannabidiol in combination with 3mg/mL of cubilose extract both significantly increased the transcript levels of FLG2 and AQP3 encoding genes. Moreover, when cannabidiol was combined at 5 μ g/mL with 3mg/mL of cubilose extract, the enhancement effect on FLG2 and AQP3 coding gene expression was best, with an enhancement rate of about 30% to 80%.

Therefore, the combination of cannabidiol and the cubilose extract can more remarkably improve the expression of the genes related to skin moisturizing, and therefore, the combination can be the best choice for applying to skin cosmetics.

Example 4: preparation of cannabidiol-bird's nest cream

To further test the safety and moisturizing properties of the topical skin application of the combination of cannabidiol and cubilose extract, the inventors prepared a cream containing cannabidiol and cubilose extract, whose raw materials include the following ingredients in parts by weight:

phase A: 66.6% of deionized water, 0.2% of carbomer (U20), 0.1% of EDTA-2 Na0.1%, 0.15% of methylparaben, 3% of glycerol, 0.2% of allantoin, 0.1% of xanthan gum and 3% of 1, 3-butanediol;

phase B: 3% of a seebeck emulsifier, 2% of triethanolamine, 1% of 16/18 alcohol, 0.5% of batyl alcohol, 0.5% of squalane, 5% of ethyl palmitate, 4% of caprylic/capric triglyceride, 5% of isononyl isononanoate, 2% of polydimethylsiloxane, 0.5% of tocopheryl acetate, 0.1% of hydroxyphenyl ester;

and C phase: 0.25% of Triethanolamine (TEA) and 1% of water;

phase D: 2% of essence, 1% of purslane and 0.5% of phenoxyethanol;

active ingredients: cannabidiol 0.0005% and nidus Collocaliae extract 0.3%.

The preparation method of the cream comprises the following steps:

1) spraying U20 on the surface of deionized water, adding other components in phase A after completely wetting, stirring at 1300rpm at about 85 deg.C for 30min until completely dissolving to obtain phase A;

2) mixing all components in phase B, heating to 80 deg.C, maintaining for 20min, adding into phase A, and stirring for two minutes for homogenization;

3) continuously stirring the mixture of the phase A and the phase B at 80 ℃ for 10min, then cooling to about 65 ℃, and carrying out vacuum defoaming;

4) adding phase C at about 60 deg.C;

5) adding phase D at about 45 deg.C;

6) and finally adding cannabidiol and the cubilose extract, stirring uniformly, and cooling to room temperature.

Example 5: skin sensitivity test

12 volunteers (6 men and 6 women, and healthy skin in the lower arm) were selected, and after being acclimated in a constant temperature (24 + -1 deg.C) and constant humidity (50 + -5%) room for 30 minutes, photographs were taken with a camera, and the skin in the test area, i.e., the left lower arm (test group) and the right lower arm (control group), was tested. First, the cannabidiol-cubilose cream prepared in example 4 was applied to the skin of the left forearm test area of the volunteer, respectively, after waiting for 1 hour, photographs were taken, and the left and right forearm were compared to observe whether there was any significant change in the skin of the test area, such as redness, itching, allergic reaction, etc., after which the same test was performed every other day and the photographs were observed and taken.

Fig. 7 shows a graph of the results of the human skin sensitivity test performed on the 12 volunteers using a cream comprising Cannabidiol (CBD) and bird's nest extract. As shown in fig. 7, no significant change was observed in the skin of all volunteers, indicating that the cream of the present invention is non-irritating to the skin, mild and has a strong moisturizing formulation.

Example 6: testing of moisturizing performance

12 volunteers (6 male and 6 female, healthy skin of forearm) were selected, first, acclimatized in a constant temperature (24 + -1 deg.C) and constant humidity (50 + -5%) room for 30 minutes, and then used

The Moisture content of the skin of the left forearm of 12 volunteers was measured by a skin Moisture meter, and the result was expressed by a Moisture Measurement Value (MMV), which was MMV1, i.e., MMV Value of the skin before cream application. The cannabidiol-cubilose cream prepared in example 4 was then applied to the skin of the left forearm test area of the volunteers three times a day and after one week again skin tests were carried out, yielding a moisture measurement MMV 2. MMV1 and MMV2 were compared to evaluate the moisturizing performance of the cream of the present invention. The test results are shown in table 1 below.

Table 1: moisturizing rate of cream of the present invention

Wherein: MMVl: skin MMV value before application of cream; MMV 2: skin MMV value 1 week after cream application; the moisturizing rate is MMV2/MMV1 multiplied by 100%.

As shown in Table 1, the results of the skin moisturizing performance test table show that the skin moisturizing rate of the cannabidiol-cubilose cream is remarkably improved after twelve volunteers use the cream for one week, and no skin allergic reaction occurs.

Claims

1. Use of cannabidiol for moisturizing skin cells by increasing gene expression of Aquaporin 3(AQP3) and/or skin outer glycoprotein Filaggrin-2(FLG2) in skin cells.

2. Use according to claim 1, wherein the cannabidiol is present in an effective concentration of 0.0001 to 0.001 wt%, preferably 0.0005 wt%.

3. Use according to claim 1 or 2, wherein cannabidiol is further used in combination with cubilose extract for skin cell moisturizing.

4. Use according to any one of claims 1 to 3, wherein the effective concentration of the cubilose extract is at least 0.01% by weight, preferably at least 0.3% by weight.

5. The use of any one of claims 1-4, wherein cannabidiol, or the combination of cannabidiol and bird's nest extract, moisturizes and moisturizes skin cells by increasing the hydration level of skin, increasing skin moisturization, and/or conditioning and moisturizing skin.

6. A composition for moisturizing skin cells comprising cannabidiol and cubilose extract.

7. The composition of claim 6 wherein cannabidiol is present in an amount of 0.0001 to 0.001% by weight of the composition, preferably 0.0005% by weight of the composition.

8. Composition according to any one of claims 6 to 7, wherein the cubilose extract is at least 0.01% by weight of the composition, preferably at least 0.3% by weight of the composition.

9. A method of increasing gene expression of Aquaporin 3(AQP3) and/or the skin outer glycoprotein filggrin-2 (FLG2) in a skin cell comprising administering cannabidiol and/or a bird's nest extract to said skin cell.

10. The method according to claim 9, wherein the cannabidiol is present in an effective concentration of 0.0001% to 0.001%, preferably 0.0005% by weight.

11. The method according to claim 9 or 10, wherein the effective concentration of the cubilose extract is at least 0.01 wt.%, preferably at least 0.3 wt.%.

12. A method of treating skin comprising topically applying the composition of any one of claims 6-8 to skin in need thereof.

13. The method of claim 12, wherein the composition is applied to skin in need of moisturizing.

14. The method of claim 12 or 13, wherein the composition is applied to striae, darkness, uneven skin tone, dryness, desquamation, laxity, and/or inelasticity of skin resulting from water deficit.