TWI448308B - Rose and rosemary extracts composition and its use for enhancing cell proliferation thereof - Google Patents

Description

Rose and rosemary extract composition and use thereof for promoting cell proliferation

The present invention relates to a plant extract composition. In particular, the invention relates to a composition comprising a rose extract and a rosemary extract.

With age or long-term exposure to harmful environmental factors, the visual appearance, physical properties and physiological functions of the skin can occur visually unpleasantly, such as fine lines and wrinkles, loss of elasticity, increased relaxation, loss of firmness, loss Color uniformity or hue, rough surface texture and plaque pigmentation. Many changes in the appearance and function of the skin are caused by changes in the outer epidermis of the skin. Less obvious but measurable changes that occur when the skin ages or is exposed to long-term environmental damage include an overall decrease in cell and tissue viability, cell replication. Reduced rate, reduced blood flow to the skin, reduced moisture content, defects in structure and function accumulation, changes in the normal regulation of common biochemical pathways, and reduced skin remodeling and ability to repair itself.

In addition, the collagen of the skin is the main force to provide skin support, and the fibroblasts in the dermis of the skin are the manufacturing plants of collagen. When young, the fibroblasts have strong activity and the collagen production is high. The mother cells not only have reduced activity, but also have fewer and fewer new cells, which makes collagen production less, the skin loses support, becomes loose and has no elasticity, and causes wrinkles.

At present, there are several methods to treat skin damage caused by aging or other factors, such as the use of agents or squares that stimulate the rate of replacement of the epidermis itself, allowing an increase in the rate of epidermal cell renewal (usually by faster rate epithelial cells) Copy to achieve); apply hyaluronic acid or collagen to maintain skin elasticity, but this method does not hold Long-lasting, expensive and invasive treatment is not high; using toner laser to reduce pores and stimulate the activity of fibroblasts in the dermis of the skin to make collagen, however, its disadvantage is the intensity of the laser If it is not properly controlled, it will easily cause skin damage.

The method of the above method has various drawbacks such as significant irritation to the skin or skin toxicity. In addition, most of these methods often irritate the skin and cause chronic damage to the skin. Therefore, it is still necessary to find a natural way to regulate and improve the skin.

Due to the defects existing in the prior art, the inventors have unexpectedly found that after extracting the rose extract and the rosemary extract in a special ratio, the effect of promoting cell proliferation can be achieved, and further Achieve conditioning and improve skin.

One aspect of the present invention provides a composition comprising an effective amount of rose extract and rosemary extract, wherein the rose extract and the rosemary extract have a weight ratio of from 10:1 to 1:10. In an embodiment of the invention, preferably, the rose extract and the rosemary extract have a weight ratio of 5:1.

The composition according to the present invention, wherein the effective amount of the rose extract and the rosemary extract is from about 1.0% to 15% by weight of the composition, such as the effective amount of the rose extract and the rosemary extract. The composition by weight ratio is 1.0%, 1.5%, 2.0%, 2.5%, 3.0%, 3.5%, 4.0%, 4.5%, 5.0%, 6.0%, 7.0%, 8.0%, 9.0%, 10%, 11% , 12%, 13%, 14% or 15%. In a preferred embodiment of the present invention, the effective amount of the rose extract and the rosemary extract The composition comprises from 2.5% to 10% by weight, most preferably 10% by weight.

According to the composition of the present invention, the extraction method of the rose extract and the rosemary extract is not limited, and an extraction method suitable for extraction such as distillation, solvent extraction or cold press extraction is applicable.

Also in the present invention, the composition may be present in any form, and is preferably in the form of an emulsion, cream, lotion, solution, anhydrous base, gel, spray or ointment.

Another aspect of the invention provides the use of a composition of the invention, wherein the composition is for promoting cell proliferation.

In a preferred embodiment, the use of the composition of the invention is for promoting proliferation of epidermal keratinocytes and/or fibroblasts.

The detailed description of the present invention and the preferred embodiments thereof will be described in the following embodiments, which are to be understood by those of ordinary skill in the art.

definition

The term "a" or "an", when used in conjunction with "including", is used in the scope of the application or the specification, which may be one, but also one or more or at least one.

The term "rose" as used herein refers to a plant of the genus Rosaceae, including but not limited to Rosa damascena , Rosa centifolia L , Rosa multiflora or Rosa gallica . Etc. And it is better to use Damascus Rose.

The term "rosemary" (scientific name: Rosmarinus officinalis ), including but not limited to Rosemary , Severn Sea Rosemary , Rex Rosemary , Vervain Rosemary ( Rosmarinus officinalis ), Pine Rosemary , Pinkie Rosemary , Blue Boy Rosemary or Lockwood Rosemary .

The term "extract" as used herein refers to the extraction of rose or rosemary from its active ingredients using a purification separation technique. Among them, the purification separation technique is not limited, and for example, a distillation method, a solvent extraction method, a cold pressing method, or the like can be used. Among them, the most common user is direct steam distillation, in which rose or rosemary is placed in a distillation column, directly or indirectly in contact with steam, boiled and vaporized, and then condensed back to liquid through a condenser to obtain an extract. Further, the aqueous phase extracting substance (or pure dew) may be separated from the oil-water separator. Another indirect steam distillation method is to heat the rose or rosemary together with water to boiling, and then vaporize and then condense back to the liquid through the condenser to obtain an extract, and further separate the water from the oil-water separator. Phase extraction material (or pure dew); solvent extraction method using alcohol, ether, liquid butane and other solvents, repeatedly dripping on the rose or rosemary, and then separating and dissolving the solvent containing the essential oil, and removing the solvent by low temperature distillation , you can get the essential oil that cures and does not flow easily. The cold pressing method extracts the rose or rosemary by filtering, analyzing and centrifuging the oil to extract the extract.

The term "promoting cell proliferation" as used herein refers to promoting the proliferation of epidermal keratinocytes and/or fibroblasts, thereby improving skin metabolism and enhancing collagen production, and improving the appearance of skin appearance. Typical improvements include (but are not limited to): skin thickness, elasticity, wetness, smoothness, hue, texture, shine, shine, brightness, transparency, contour, firmness, firmness, smoothness, softness, Sensitivity, pore size, or desired characteristics and/or properties associated with such combinations. These terms are also used to mean an improvement in an undesirable skin condition. Typical adverse conditions caused, caused or caused by such undesirable skin conditions include (but are not limited to): dryness, eczema, liposuction dermatitis, dermatitis, sunburn, estrogen imbalance, pigmentation Degree of deposition, light pigmentation, discoloration, yellowing, freckles, skin atrophy, skin loss, skin fragility, dryness, rough touch, cracking, sagging, thin skin, hyperplasia, fibrosis, increased pores, honeycomb Tissue formation, bruising, formation of acne, apoptosis, cell differentiation, cell differentiation, induction of cancer or cancer spread, viral infection, fungal infection, bacterial infection, venous flexion, hirsutism, rosacea, itching Symptom or a combination of these.

One aspect of the present invention provides a composition comprising an effective amount of rose extract and rosemary extract, wherein the rose extract and the rosemary extract have a weight ratio of from 10:1 to 1:10. The weight ratio of the extractable rose extract to the rosemary extract may be 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 1:5, 1:6, 1 : 7, 1:8, 1:9, 1:10, and in a preferred embodiment of the invention, the rose extract and the rosemary extract have a weight ratio of 5:1.

In the composition of the present invention, wherein the effective amount of the rose extract and the rosemary extract is from about 1.0% to 15% by weight of the composition, such as the effective amount of the rose extract and the rosemary extract. The composition by weight ratio is 1.0%, 1.5%, 2.0%, 2.5%, 3.0%, 3.5%, 4.0%, 4.5%, 5.0%, 6.0%, 7.0%, 8.0%, 9.0%, 10%, 11% , 12%, 13%, 14% or 15%. In a preferred embodiment of the invention, the effective amount of the rose extract and the rosemary extract is from 2.5% to 10%, most preferably 10% by weight of the composition.

Another aspect of the invention provides the use of a composition of the invention, wherein the composition is for promoting cell proliferation. In a preferred embodiment, the use of the composition of the invention is for promoting proliferation of epidermal keratinocytes and/or fibroblasts.

Based on the foregoing description, the present invention provides a composition comprising an effective amount of rose extract and rosemary extract that can be applied to a skin care product. This skin care product can be applied to cosmetics or pharmaceuticals, including daily skin care. This composition may further comprise Other ingredients which are selected from a cosmetically acceptable carrier or a pharmaceutically acceptable carrier. This product type contains aqueous solutions, semi-solids and solids; such as lotions, lotions, masks, gels, creams, balms, ointments, liquids, powders or others that can be used for topical or internal use. type.

The carrier which can be used in the composition of the present invention is a cosmetic or pharmaceutically acceptable carrier, and refers to a non-toxic carrier which is conventionally used for cosmetic or pharmaceutical use, and can be more convenient for use on cosmetics or medicines, and It maintains the stability and bioavailability of rose extract and rosemary extract when applied directly to the skin surface. For example, the rose extract and the rosemary extract are dissolved in an aqueous solution, dispersed or emulsified in a matrix, prepared as an aqueous solution depending on the convenience of use, or mixed with a semi-solid (gelatin) or a solid carrier to form a gel. , powder, paste, lotion, lotion or the like. Suitable cosmetically or pharmaceutically acceptable carriers are the same as those known to those of ordinary skill in the art, including cosmetically or pharmaceutically acceptable liquids, creams, oils, lotions, lotions, gels. Or solid forms include cosmetic night creams, foundation creams, sunscreen creams, sunscreen lotions, hand lotions, foundations and foundations, masks and the like. This composition may contain other therapeutically beneficial ingredients such as epithelial cell growth factor (EGF), platelet derived growth factor (PDGF), or transformed cell growth factor (TGF-alpha). On the other hand, this composition may contain other ingredients conventionally added to cosmetics, such as essence, estrogen, vitamin A, C, E and, lactic acid or glycolic acid, lanolin, aloe, methylation or Propane-based preservatives, pigments and the like. Suitable cosmetic or pharmaceutically acceptable carriers include water, petrolatum, petrolatum oil, mineral oil, vegetable oil, animal oil, organic and inorganic waxes, such as microcrystalline waxes, paraffin waxes, ceresin waxes; natural polymers like xanthanes, several Butadiene, cellulose, collagen, starch and gum arabic, synthetic polymers like alcohols, polyols and their analogues as discussed below. Because the non-toxic body contains The body traits are mostly composed of a carrier that is easy to mix with water, such as a host cosmetic or pharmaceutically acceptable carrier that is easily miscible with water, and includes not only one or more of the other suitable ingredients, but also may be maintained or A carrier that slows the release rate, including hydrates, water-dispersible or water-soluble components such as vesicles, micro-sponges, microparticles, microcapsules, aqueous ointments, water-soluble oils or oil-soluble emulsions, Gel or analogue thereof. If expected, one or more additional ingredients are usually found in the main cosmetic composition, including the types of carriers such as moisturizing lotion, moisturizer, fragrance, buffer, pigment, preservative, vitamin ACE, emulsifier. , dispersants, wetting agents, perfumes, colloids, stabilizers, coatings, antibacterial agents, sunscreen ingredients, enzymes and the like. Those skilled in the art can select the appropriate specific additions, including the type and quantity.

Liposomes can also be added to the compositions of the present invention as carriers for assisting in the delivery of the active substance within the composition into the dermal layer of the skin. Other dermal layer delivery vehicles which may be substituted for liposomes include, but are not limited to, those known to those skilled in the art to include a skin delivery vehicle comprising: an emulsifier, a particulate emulsifier, a nano emulsifier. , nanoparticle, microspheres, hydrophilic and lipophilic vesicles (ethosome), transfer vesicles, and nonionic surfactant vesicles (niosomes).

The composition of the present invention may optionally comprise one or more anti-allergic agents, anti-discomfort agents, anti-fungal agents, antimicrobial agents, anti-inflammatory agents, sterilizing agents, chelating agents, coloring agents, pigment removing agents, softening agents (such as Triglyceride butyrate, citric acid/caprate, castor oil, ceteareth 20, ceteareth-30, cetyl stearyl alcohol, cetyl alcohol, cetyl stearyl alcohol, cholesterol, cocoa butter, Isopropyl diacrylate, glycerin, glycerol monooleate, glyceryl monostearate, glyceryl stearate, isoparaffin, isopropyl myristate, isopropyl palmitate, lanolin, lanolin alcohol , hydrogenated lanolin, isoparaffin, linoleic acid, mineral oil, oleic acid, petrolatum, phosphorus Lipid, polyethylene glycol, polyethylene glycol-7-cocoa oleic acid glyceride, polyethylene glycol-18-methyl ester dimethyl decane, polyethylene glycol fatty alcohol ether, polyglycidyl propylene glycol 15 - stearyl ether, polypropylene glycol propylene glycol, polypropylene glycol stearate, sorbitol, sphingolipid, squalene, steareth-2 or -100 stearic acid, stearyl alcohol, urea, petrolatum or the like ), emulsifier, film former, perfume, wetting agent (ethyl glucosamine, disaccharide gum, butanediol, ethoxy diethylene glycol, ethylene glycol polypropylene, glucose, glycereth-26, glycerin) , glycerol, ethylene glycol, lactitol, maltitol, propylene glycol, sericin, sodium hyaluronate, sorbitol, xylitol, sodium citrate, glucose or the like, and insect repellent , lubricants, humectants, pharmaceutical agents, photosensitizers, preservatives (such as ethylenediaminetetraacetic acid (EDTA) dipotassium salt, benzyl alcohol, methyl paraben or isobutyl p-hydroxybenzoate), Skin protectant, skin penetration enhancer, sunscreen, stabilizer, surfactant, thickener (such as xanthan gum, carrageenan, Ming , Karaya, pectin or thorn Gui bean gum), viscosity modifier, or a combination of these.

The cosmetic composition of the present invention may further comprise an additional cosmetic ingredient comprising, but not limited to, the following ingredients: licorice, licorice extract, licorice derivative (for example: glycyrrhizinate); lemon extract; Cucumber extract; sunflower seed extract; castor seed oil; oat protein, oat extract, hydrolyzed oatmeal; silk protein (eg sericin); hyaluronic acid and its derivatives (eg sodium hyaluronate); vitamin Minerals; antioxidants; phospholipids, sphingolipids, cholesterol; and/or other ingredients that have anti-aging, anti-oxidant, anti-inflammatory, anti-discomfort, anti-cancer, or other skin-protecting properties or combinations thereof.

The frequency of application of the compositions of the invention to the skin can be at least once a day. When the cosmetic composition of the present invention is applied once a day, it is preferably applied at night to reduce additional environmental damage. The composition of the present invention can effectively provide for promoting the proliferation of epidermal keratinocytes and/or fibroblasts during a period of administration, thereby improving the appearance. In a specific example, the composition of the present invention It is administered at least once a day for a period of 14 days or longer.

The composition of the present invention can be contained in a container according to its suitability. For example, a lotion or a cream can be contained in a bottle, and if it is liquid, it can be contained in a ball feeder, or a pusher to drive the spray device, and a device suitable for finger operation. Pu's container or glass bottle in the dark. When the composition of the present invention is a cream, it can be simply stored in a non-deformable bottle or squeezable container. Thus, the invention may also be a closed container containing a cosmetically acceptable composition as defined herein.

The present invention also provides a kit suitable for providing a composition dose for a desired period of time (e.g., 14 days) during a therapeutic procedure. Referring to this specific example, a set consists of a package consisting of several vials, each containing a suitable dose for a single application of the cosmetic composition of the present invention. In one particular embodiment, a set comprises a package of 14 vials, each containing a composition dose sufficient for a single application of the skin.

Example

Hereinafter, the present invention will be further described in detail with reference to the embodiments. However, it is to be understood that these examples are only intended to aid in the understanding of the invention and are not intended to limit the scope of the invention.

Production Example 1 - Extracting Rose Extract

Using direct steam distillation, one kilogram of fresh Damascus rose petals are placed on a rack in a distillation column. Water is injected into the lower end of the distillation column and heated to above 100 ° C to evaporate water to produce water vapor. The water vapor is passed from the pipe in the distillation column to the pipe. Rose petals, after destroying the petal tissue, drive the extracted material through the cooling system, and directly obtain the liquid obtained by condensation, which is a rose extract with a yield of about 1 c.c.

Production Example 2 - Extraction of Rosemary Extract

In the same manner as in Production Example 1, only Damascus rose petals were replaced with verbena ketone rosemary leaves. The rest of the conditions are the same and the yield is about 0.6 c.c.

Example 1 - Effect of the 2.5% concentration of the present invention on the growth of embryonic fibroblasts and epidermal keratinocytes

After the rose extract and the rosemary extract were mixed at a weight ratio of 5:1, the mixture and the ultrapure water were blended as shown in Table 1 below to a dilution containing a mixture concentration of 5%. Embryonic fibroblasts NIH3T3 and human epidermal keratinocyte strain HaCaT, which are commercially available from mice, were separately studied.

NH3T3: Place NIH3T3 in Dulbecco's Modified Eagle Medium (DMEM) mixed culture medium containing 10% fetal bovine serum and 1% antibiotic, place it evenly into a 75 T-flat flask, and place it in a 37 °C cell culture incubator. Cultured in (5% CO ₂ ), after the cells were about 9 minutes full, wash twice with PBS, add 5 ml of 0.5% trypsin-EDTA (GIBCO), and place at 37 ° C, 5% CO ₂ incubator In the middle, the effect is about 5 minutes. After observing the cells under the microscope, the effect of trypsin was added to the culture solution containing fetal bovine serum. The mixed cell liquid was placed in a 15 ml centrifuge tube, centrifuged at 1500 rpm for 5 minutes, and after centrifugation, the supernatant was aspirated, leaving the cells deposited at the bottom of the tube, and the cells were counted to adjust the cell volume to 2 × 10 ⁵ / Ml, into a cell suspension; the above cell suspension is mixed with rose extract and rosemary extract at a concentration of 2.5% of the mixture at a concentration of 1:1 (the concentration of rose extract and rosemary extract at this time is 2.5%), and in a 96-well plate, each well was inoculated with 100 μl, and the 96-well plate was placed in a 37 ° C, 5% CO ₂ incubator. After 24 and 48 hours of culture, 10% of the culture solution was added. The volume of WST-1 reagent was measured for two hours. The absorbance of OD440nm was measured by an enzyme immunoassay analyzer, and the absorbance of the readout was compared with the control group to evaluate the cell survival rate. Each experiment of the same conditions was repeated three times, the average was taken, and t-test was used as a statistical method.

HaCaT: The experimental method was the same as that of NIH3T3, and only the cells were replaced with HaCaT.

Cell viability is shown in Table 2 and Figures 1 and 2.

Example 2 - Effect of the concentration of 10% mixture of the present invention on the growth of embryonic fibroblasts and epidermal keratinocytes

After the rose extract and the rosemary extract were mixed at a weight ratio of 5:1, the mixture and the ultrapure water were blended as shown in Table 1 below to a dilution containing a mixture concentration of 20%. The rose extract and the rosemary extract used in Example 1 were replaced with a dilution of 5% by weight of the mixture into a dilution of 20% of the mixture prepared as above (after mixing with the cell suspension 1:1) The concentration of rose extract and rosemary extract was 10%), and the other experimental methods and cells used were the same as in Example 1.

Cell viability is shown in Table 2 and Figures 1 and 2.

Comparative Example 1 - Effect of 2.5% Rose Extract on the Growth of Embryonic Fibroblasts and Epidermal Keratinocytes

The rose extract and ultrapure water were blended as shown in Table 1 below to a dilution containing 5% rose extract. The rose extract and the rosemary extract used in Example 1 were replaced with a dilution of 5% of the mixture concentration into a dilution of 5% of the rose extract concentration as described above (mixed with the cell suspension 1:1) At this time, the concentration of the rose extract was 2.5%), and the other experimental methods and cells used were the same as in Example 1.

Cell viability is shown in Table 2 and Figures 1 and 2.

Comparative Example 2 - Effect of 10% Rose Extract on the Growth of Embryonic Fibroblasts and Epidermal Keratinocytes

The rose extract and ultrapure water were blended as shown in Table 1 below to a dilution containing a rose extract concentration of 20%. The rose extract and the rosemary extract used in Example 1 were concentrated in a mixture The 5% dilution was replaced with a 20% dilution of the rose extract as described above (after mixing with the cell suspension 1:1, the concentration of the rose extract was 10%), and the rest of the experimental methods and use The cells were the same as in Example 1.

Cell viability is shown in Table 2 and Figures 1 and 2.

Comparative Example 3 - Effect of 2.5% concentration of rosemary extract on the growth of embryonic fibroblasts and epidermal keratinocytes

The rosemary extract and ultrapure water were formulated as shown in Table 1 below to a dilution containing a rosemary extract concentration of 5%. The rose extract and the rosemary extract used in Example 1 were replaced with a dilution of 5% of the mixture concentration to a 5% dilution of the rosemary extract as described above (1:1 with the cell suspension). After mixing, the concentration of the rosemary extract was 2.5% at this time, and the other experimental methods and cells used were the same as in Example 1.

Cell viability is shown in Table 2 and Figures 1 and 2.

Comparative Example 4 - Effect of 10% concentration of rosemary extract on the growth of embryonic fibroblasts and epidermal keratinocytes

The rosemary extract and ultrapure water were formulated as shown in Table 1 below to a dilution containing a rosemary extract concentration of 20%. The rose extract and the rosemary extract used in Example 1 were replaced with a dilution of 5% of the mixture to a dilution of 20% of the rosemary extract as described above (with a cell suspension of 1:1). After mixing, the concentration of the rosemary extract was 10% at this time, and the other experimental methods and cells used were the same as in Example 1.

Cell viability is shown in Table 2 and Figures 1 and 2.

Figure 1 is a graph showing the effect of the composition of the present invention and the comparative example on the growth of embryonic fibroblast NIH3T3 at different concentrations. As can be seen from Table 2 above, the rose and rosemary extract mixture of the present invention was used. The use of rose extract or rosemary extract is statistically significant for promoting the growth of embryonic fibroblast NIH3T3, and the use of the composition of the present invention is indeed more effective than the use of only rose extract or rosemary extract. .

2 shows the effect of the composition of the present invention and the comparative example on the growth of the human epidermal keratinocyte strain HaCaT at different concentrations. As shown in Table 2, it can be seen that the rose and rosemary extract mixture of the present invention is used. There is a statistical difference in promoting the growth of human epidermal keratinocyte strain HaCaT by using only rose extract or rosemary extract. It is true that the use of the composition of the present invention is more than the use of rose extract or rosemary extract. Come to be effective.

Therefore, in summary, the composition containing the rose extract and the rosemary extract of the present invention can achieve the effect of promoting cell proliferation, and can be applied to various products.

Although the embodiments are disclosed herein, it is to be understood that the scope of the present invention is not to be construed as a For the scope of the patent application below.

Figure 1 is a graph showing the effects of the composition of the present invention and the comparative examples on the growth of embryonic fibroblasts NIH3T3 at different concentrations.

Figure 2 is a composition of the present invention and a comparative example at different concentrations on a human scale The effect of the growth of the keratinocyte strain HaCaT.

Claims (10)

A composition comprising an effective amount of rose extract and rosemary extract, wherein the rose extract and the rosemary extract have a weight ratio of from 10:1 to 1:10. The composition of claim 1, wherein the rose extract and the rosemary extract have a weight ratio of 5:1. The composition of claim 1, wherein the rose extract and the rosemary extract are present in an effective amount of from about 1.0% to about 15% by weight of the composition. The composition of claim 1, wherein the rose extract is extracted from rose petals, and wherein the rosemary extract is extracted from rosemary leaves. The composition of any one of claims 1 to 4, wherein the rose extract and the rosemary extract are extracted by distillation, solvent extraction or cold pressing. The composition of claim 5, wherein the composition is in the form of an emulsion, a cream, a lotion, a solution, an anhydrous base, a gel, a spray or an ointment. A use of the composition of claim 1 for the manufacture of a medicament for promoting cell proliferation. The use of claim 7, wherein the rose extract and the rosemary extract have a weight ratio of 5:1, and wherein the effective amount of the rose extract and the rosemary extract is about 1.0 by weight of the composition. % to 15%. For the use of claim 7, the rose extract is extracted from rose petals, and wherein the rosemary extract is extracted from rosemary leaves. The use of claim 7, wherein the rose extract and the rosemary extract are extracted by distillation, solvent extraction or cold pressing.