TWI767168B - Use of rosa plant extract for improving mitochondrial activity - Google Patents

Abstract

The present disclosure provides a use of a Rosa plant extract for anti-aging and anti-oxidation.

Description

Use of rose plant extract for enhancing mitochondrial activity

The present invention relates to the use of a rose plant extract for anti-aging and anti-oxidation.

The skin is the largest barrier to protect the human individual, it has the function of resisting water loss, pathogenic bacteria and various environmental damages. Exposure to large amounts of 3C blue light (such as mobile phones and tablets), ultraviolet (ultraviolet, UV), ionizing radiation (ionizing radiation), drugs or xenobiotics (xenobiotics) can stimulate the skin to generate reactive oxygen species (ROS) and free radicals. When the amount of accumulated reactive oxygen species and free radicals exceeds the antioxidant capacity of cells or tissues, oxidative stress is formed. Next, reactive oxygen species and free radicals react with intracellular constituents (including DNA, proteins, lipids, etc.), thereby having undesired effects on the skin.

Melanogenesis (ie, melanin synthesis) refers to when dermal melanocytes are exposed to environmental factors (such as blue light, ultraviolet (UV)) or physiological factors (such as fatigue, After the induction of stress, chronic inflammation, and abnormal α-melanocyte stimulating hormone (α-MSH) release in the body, tyrosine ( tyrosine) is converted into melanin via tyrosinase catalysis (which is the rate-limiting step of melanin production) and a series of redox reactions. Melanin can protect the hypodermis of the skin from photodamage caused by ultraviolet rays, but when melanin is accumulated in large amounts on the skin or distributed abnormally, it may lead to skin diseases (skin). disorders such as lentigines, freckles, melasma, age spots and hyperpigmentation.

In recent years, the demand for inhibiting melanin production and reducing the production of free radicals in cells is increasing day by day, because once the production of melanin is inhibited and the production of free radicals in cells is reduced, anti-aging and anti-oxidative effects can be achieved. However, the most common methods currently used to inhibit melanin production and reduce free radical production in cells are to use cosmetics, skin care products applied to the skin surface, or oral health foods that claim to inhibit melanin production and reduce free radical production in cells. However, conventional cosmetics, skin care products and health foods are mostly made of chemical ingredients. Long-term use is not only harmful to human health, but also these products are often expensive and not affordable for ordinary users.

On the other hand, the mitochondria (mitochondria) is also known as the power station of the cell, because it is the main site for the synthesis of adenosine triphosphate (ATP) (a molecule that transmits energy) in the cell, providing various activities of the cell. chemical energy. If mitochondria are damaged, the impact on cells and individual organisms is huge. During the process of synthesizing ATP, mitochondria will generate a lot of free radicals. The activity of free radicals is extremely strong, and they will have strong oxidation reactions with any substances in the body and destroy their normal functions. Free radicals damage enzymes and DNA in mitochondria over time, gradually reducing their function and thus the function of various organs and tissues. Therefore, how to enhance the mitochondrial activity of cells to achieve anti-aging and anti-oxidative effects has become an important topic in the field.

In order to solve the above problems, those skilled in the art urgently need to develop novel medicines, food products or skin care products with the functions of enhancing mitochondrial activity of cells, reducing free radical production in cells, inhibiting melanin production, anti-aging and anti-oxidation. In order to benefit the vast number of ethnic groups in need.

In view of this, the object of the present invention is to provide a rose ( ROSA ) plant extract for preparing an anti-aging and anti-oxidant composition, wherein the rose plant extract is extracted with a solvent from a rose plant As obtained, the solvent is water, glycerol, aqueous glycerol or a combination thereof, wherein the rose plant extract is a rose ( Rosa rugosa ) flower extract.

In one embodiment of the present invention, the anti-aging and anti-oxidation include enhancing mitochondrial activity of cells, reducing free radical production in cells and inhibiting melanin production.

In one embodiment of the invention, the cell is a dermal fibroblast.

In one embodiment of the present invention, the free radical generation is caused by a blue light or a hydrogen peroxide.

In one embodiment of the present invention, the melanin production is caused by a blue light.

In one embodiment of the present invention, the effective concentration of the rose extract is at least 0.25% (v/v).

In an embodiment of the present invention, the extraction is performed at a temperature ranging from 65°C to 85°C.

In an embodiment of the present invention, the volume ratio of the solvent to the rose is between 5-20:1-5.

In an embodiment of the present invention, the composition is a medicine, a food product or a skin care product.

In one embodiment of the present invention, the pharmaceutical product comprises a pharmaceutically acceptable carrier.

To sum up, the efficacy of the rose plant extract of the present invention is: by increasing the mitochondrial activity of cells (such as skin fibroblasts), reducing the generation of free radicals in cells (such as skin fibroblasts) (such as by Blue light) and inhibit melanin production (for example, caused by 3C blue light products such as mobile phones and tablet computers), to protect skin cells, strong anti-oxidation and anti-aging effects, thereby maintaining skin youthful vitality.

The embodiments of the present invention will be further described below. The following examples are used to illustrate the present invention, but not to limit the scope of the present invention. Anyone who is familiar with this technique, without departing from the spirit and scope of the present invention, Some changes and modifications can be made, so the protection scope of the present invention should be determined by the scope of the appended patent application.

Fig. 1 is a data graph of the efficacy of the rose plant extract of the present invention in enhancing the mitochondrial activity of skin fibroblasts, wherein "***" means p < 0.001 compared with the control group.

Figure 2 is a data graph of the efficacy of the present rose plant extract in reducing the generation of free radicals in cells caused by blue light, wherein "**" means p < 0.01.

Figure 3 is a data graph of the efficacy of the present rose plant extract in reducing the generation of free radicals in cells caused by hydrogen peroxide.

Figure 4 is a data graph showing the efficacy of the rose plant extract of the present invention in inhibiting blue light-induced melanin production.

definition

Numerical values used herein are approximations and all experimental data are expressed within 20%, preferably within 10%, and most preferably within 5%.

Statistical analysis was performed using Excel software. Data were expressed as mean ± standard deviation (STDEV), and differences between them were analyzed by Student's t -test.

The plant material Rosa rugosa used in the present invention is a plant of the genus Rosa of the family Rosaceae. Tian Rucheng in Ming Dynasty, "Traveling the West Lake". "Weixiang Cong Tan" mentioned that "roses mixed with brain musk deer are considered sachets, and the fragrance is endless", so it is also known as wandering flower, which is said to have a long and lingering fragrance and is loved by people.

As used herein, the term "anti-aging" means preventing, slowing down the appearance of aging phenomena of human skin, such as the development of wrinkles and loss of elasticity. The extent to which this is assessed will depend upon a number of factors known to those skilled in the art, such as the general condition of the consumer, age, gender, and the like.

As used herein, the terms "inhibition of melanogenesis" and "inhibition of melanin synthesis", "depigmenting", "lightening the melanin", " Whitening", "skin color lightening", "bleaching", "whitening", "brightening", "blackening" and "blackening" can be used interchangeably .

In accordance with the present invention, medicinal products can be manufactured into a product suitable for parenterally, orally or topically administration using techniques well known to those skilled in the art dosage forms, including, but not limited to: injections [eg, sterile aqueous solutions or dispersions], sterile powders, tablets, tablets Troche, lozenge, pill, capsule, dispersible powder or granule, solution, suspension, emulsion, syrup (syrup), elixir (elixir), slurry (slurry), external preparation (external preparation) and the like.

According to the present invention, the pharmaceutical product may further comprise a pharmaceutically acceptable carrier which is widely used in pharmaceutical manufacturing technology. For example, the pharmaceutically acceptable carrier may comprise one or more agents selected from the group consisting of: solvent, buffer, emulsifier, suspending agent, decomposer ), disintegrating agent, dispersing agent, binding agent, excipient, stabilizing agent, chelating agent, diluent , gelling agents, preservatives, wetting agents, lubricants, absorption delaying agents, liposomes, and the like. The selection and quantity of these reagents are within the scope of the expertise and routine skills of those skilled in the art.

According to the present invention, the pharmaceutically acceptable carrier comprises a solvent selected from the group consisting of water, normal saline, phosphate buffered saline (PBS), Aqueous solutions containing alcohol and combinations thereof.

According to the present invention, the medicinal product may be administered by a parenteral route selected from the group consisting of: intraperitoneal injection, subcutaneous injection, intradermal injection Intraepidermal injection, intradermal injection, intramuscular injection, intravenous injection and intralesional injection.

In accordance with the present invention, the medicinal product may be manufactured into an external preparation suitable for topical application to the skin using techniques well known to those skilled in the art, including, but not limited to: emulsions, gels Gel, ointment, cream, patch (patch), liniment (liniment), powder (powder), aerosol (aerosol), spray (spray), lotion (lotion), serum (serum), paste (paste), foam (foam), drops (drop), suspension (suspension), ointment (salve) and bandage (bandage).

According to the present invention, the external preparation is prepared by mixing the medicinal product of the present invention with a base well known to those skilled in the art.

According to the present invention, the substrate may comprise one or more additives selected from the group consisting of water, alcohols, glycols, hydrocarbons (such as petroleum, jelly) and white petrolatum], waxes [such as paraffin and yellow wax], preserving agents, antioxidants, surfactants, absorption enhancers ( absorption enhancers), stabilizing agents, gelling agents [such as Carbopol ^® 974P (carbopol ^® 974P), microcrystalline cellulose and carboxymethylcellulose], Active agents, humectants, odor absorbers, fragrances, pH adjusting agents, chelating agents, emulsifiers, occlusive agents occlusive agents, emollients, thickeners, solubilizing agents, penetration enhancers, anti-irritants, colorants and propellants (propellants) and so on. The selection and quantity of these additives are within the professional and routine skills of those skilled in the art.

According to the present invention, the skincare product may further comprise an acceptable adjuvant which is widely used in skincare product manufacturing techniques. For example, the acceptable adjuvant may contain one or more agents selected from the group consisting of solvents, gelling agents, active agents, preservatives, antioxidants, screening agents, chelating agents, surfactants, dyes Coloring agents, thickening agents, fillers, fragrances and odor absorbers. The selection and quantity of these reagents are within the scope of the expertise and routine skills of those skilled in the art.

According to the present invention, the skin care product can be manufactured into a form suitable for skincare or makeup using techniques well known to those skilled in the art, including, but not limited to: aqueous solution, water -Aqueous-alcohol solution or oily solution solution), oil-in-water type, water-in-oil type or complex emulsion, gel, ointment, cream, mask, patch , pack, liniment, powder, aerosol, spray, lotion, serum, paste, foam, dispersion, drops, mousse, sunblock, tonic water ), foundation, makeup remover products, soap, and other body cleansing products.

According to the present invention, the skin care product may also be used in combination with one or more external use agents with known activity selected from the group consisting of whitening agents (such as tretinoin, catechins) (catechin), koji acid, arbutin and vitamin C], humectants, anti-inflammatory agents, bactericides, ultraviolet absorbers, plant extracts[ such as aloe extract], skin nutrients, anesthetics, anti-acne agents, antipruritics, analgesics, anti-inflammatory agents antidermatitis agents, antihyperkeratolytic agents, anti-dry skin agents, antipsoriatic agents, antiaging agents, antiwrinkle agents, Antiseborrheic agents, wound-healing agents, corticosteroids and hormones. The selection and quantity of these topical preparations fall within the scope of the professionalism and routine skills of those skilled in the art.

According to the present invention, the food product can be regarded as a food additive, which is added during the preparation of raw materials by conventional methods, or added during the production process of the food, and is formulated with any edible material for Food products consumed by humans and non-human animals.

According to the present invention, types of food products include, but are not limited to, beverages, fermented foods, bakery products, health foods, and dietary supplements.

Example 1. Preparation of Rosa ( ROSA ) Plant Extract

First, the rosa ( ROSA ) plant (preferably Rosa rugosa ) (purchased from the dried rose material produced in Iran from Xuanyang Industrial Co., Ltd.) is homogenized, and then at 65-85 ° C, the A crude extract is obtained by extracting the homogenized Rosa plant (with a volume ratio of 5~20:1~5) with a solvent, wherein the solvent is water, glycerol, water-containing glycerol or a combination thereof. The crude extract is filtered to obtain a filtrate, which is then centrifuged, and the supernatant is collected and filtered to obtain a rose plant extract.

Example 2. Evaluation of the efficacy of rose plant extracts in enhancing mitochondrial activity in cells

In this example, human skin fibroblasts CCD-966SK were used to analyze the mitochondrial activity of skin cells, and flow cytometry Mitochrondrial membrane potential detection kit (BD) was used to conduct experiments. Human skin fibroblasts were purchased from Taiwan Bioresource Collection and Research Center (BCRC), number BCRC 60153. The cells were cultured in supplemented with 10% fetal bovine serum (FBS) (GIBCO, No. 10438-026, USA), 0.1 mM non-essential amino acids, 1.5 g/L sodium bicarbonate (Sigma, No. Minimum essential medium (MEM) (Eagle) (Earle's Balanced Salt Solution, Earle's BSS)) (GIBCO Company, No. 41500-034, USA).

¹ x 105 human dermal fibroblasts (n=3) were seeded per well in a 6-well culture dish containing 2 mL of the above medium. After that, the cells were divided into two groups, including the control group and the experimental group. 0.5% (v/v) rose plant extract was added to the cells of the experimental group, and to the cells of the control group, the medium was added. Next, each group of cells was cultured at 37°C for 24 hours. Before using the BD ^TM MitoScreen(JC-1) kit, pre-warm 10x JC-1 staining assay buffer (JC-1 10x assay buffer) at 37°C, and pre-warm it with sterile 1x phosphate buffer. The solution (1x Phosphate buffered saline, 1x PBS) was prepared with 1x JC-1 staining assay buffer, and the temperature was maintained at 37 °C, and then 130 μL of dimethyl sulfoxide (DMSO) was added to the 1x JC-1 staining assay. Freeze-drying in the buffer is the storage solution of the JC-1 staining assay buffer, which can be stored at -20°C for 6 months; when it is to be used, mix the JC-1 stain with the 1x JC-1 assay buffer. The liquid is uniform in the ratio of 1:100, that is, the JC-1 dye. Then, after adding trypsin/EDTA for 3 minutes, the suspended cells were pipetted into a 1.5 mL microcentrifuge tube, and the precipitated cells were collected by centrifugation at 400 g for 5 minutes.

After removing the supernatant, cells were resuspended in 1 mL of IX PBS, then transferred to a 1.5 mL centrifuge tube and centrifuged at 400 g for 5 min. After removing the supernatant, add 100 μL of the JC-1 working solution, mix well, and act in the dark for 15 minutes. Afterwards, centrifuge at 400g for 5 minutes, wash with 1 mL of 1X wash buffer and centrifuge at 400g for 5 minutes, then wash with 1 mL of 1X wash buffer and centrifuge at 400g for 5 minutes. Cells were resuspended with 500 μL of 1X PBS containing 2% FBS, and then the mitochondrial membrane potential changes during apoptosis were observed by flow cytometry (Beckman) analysis, and t -test (student t -test) statistics were performed with Excel Statistical significance of differences between sample populations was analyzed.

Figure 1 is a data graph showing the efficacy of the rose plant extract of the present invention in enhancing mitochondrial activity in skin fibroblasts. As can be seen from Figure 1, compared with the control group, the mitochondrial activity of the skin fibroblasts in the experimental group (ie, the relative JC-1 aggregate (aggregate)) was significantly improved. Compared with the control group, the experimental group The mitochondrial activity of skin fibroblasts increased by 55.4%. The results of this example show that the rose plant extract of the present invention can significantly enhance the activity of mitochondria, maintain the youthful vitality of the skin, and achieve the potential of anti-aging.

Example 3. Evaluation of the efficacy of rose plant extracts in reducing the generation of free radicals in cells induced by blue light

First, with Dulbecco's Modified Eagle's Medium (DMEM) and Ham 's F12 supplemented with 10% fetal bovine serum (Gibco), 0.5 mM sodium pyruvate (sodium pyruvate) and 15 mM HEPES Human dermal fibroblasts CCD-966SK (BCRC 60153) were cultured in medium (1:1 mix) in a 6-well dish, and the cell concentration of 2 mL of medium was 1.5×10 ⁵ cells/well, and then cultured at 37°C for 24 hours. and remove the medium. Next, the cultured cells were divided into three groups, including a control group, a blue light group, and an experimental group. 0.5% (v/v) rose plant extract was added to the cells of the experimental group, and blue light (brand: ZAMI STUDIO, blue light wavelength: 415 nm; dose 50 J/cm ² ) was irradiated for 15 minutes. The cells in the blue light group were irradiated with blue light for 15 minutes, while the cells in the control group were not supplemented with rose plant extract and were not irradiated with blue light. After culturing cells in each group at 37°C for 1 hour, 5 μg/mL dichloro-dihydro-fluorescein diacetate (DCFH-DA) (Sigma/SI-D6883-50MG) (stock solution) was added. 5 mg/mL in DMSO) and reacted at 37°C for 15 minutes. Next, each well was washed twice with 1 mL of 1X PBS (Gibco), then 200 μL of trypsin was added and reacted in a dark environment for 5 minutes, then the cell culture was collected into a 1.5 mL volume centrifuge tube, and the Centrifuge at 400 x g for 10 minutes. After that, the supernatant was removed and washed once with IX PBS, followed by centrifugation at 400 xg for 10 minutes. Next, the supernatant was removed and the cell pellet was resuspended in 1 mL of IX PBS. After that, the fluorescence signal of DCFH-DA was detected by flow cytometer (Beckman) with excitation wavelength (450~490nm) and emission wavelength (emission wavelength) 510~550nm, thereby calculating the damage caused by ROS percentage of cells. Statistically significant differences between groups were determined by Student's t-test. The results of this example are shown in FIG. 2 .

Figure 2 is a data graph showing the efficacy of the present rose plant extract in reducing the generation of free radicals in cells caused by blue light. As can be seen from Figure 2, compared with the control group, the percentage of cells damaged by ROS (i.e. cells with high oxidative stress) measured in the blue light group was significantly increased, which means that blue light will cause a large amount of ROS damage to skin fibroblasts; Compared with the blue light group, the percentage of cells damaged by ROS in the experimental group was significantly reduced (reduced ROS expression caused by blue light by about 21%). The results of this example show that the rose plant extract of the present invention has the effect of reducing the generation of free radicals in cells caused by blue light.

Example 4. Evaluation of the efficacy of rose plant extracts in reducing the generation of cellular free radicals caused by hydrogen peroxide

First, a minimal essential medium ( MEM) (Eagle) (in Earle's Balanced Salt Solution, Earle's BSS)) (GIBCO Company, No. 41500-034, USA) cultured human dermal fibroblasts CCD-966SK (BCRC 60153) on 6- Well plates, 2 mL of medium at a cell concentration of 2 x 10 ⁵ cells/well, were then cultured at 37°C for 24 hours, and the medium was removed. Next, the cultured cells were divided into three groups, including a control group, a hydrogen peroxide group, and an experimental group. 0.25% (v/v) rose extract and ₁ mM H2O2 (Sigma) _were added to the cells of the experimental group. Cells in the hydrogen peroxide group were supplemented with 1 mM H ₂ O ₂ , while cells in the control group were left untreated. After that, 5 μg/mL dichloro-dihydro-fluorescein diacetate (DCFH-DA) (Sigma/SI-D6883-50MG) (stock solution of 5 mg/mL in DMSO) was added and added to Reactions were performed at 37°C for 15 minutes, then cells _were treated with H2O2 for ₁ hour at 37°C, followed by washing each well twice with 1 mL of IX PBS (Gibco). Next, 200 μL of trypsin was added and reacted in a dark environment for 5 minutes, then the cell culture was collected into a 1.5 mL volume centrifuge tube, and centrifuged at 400×g for 10 minutes. After that, the supernatant was removed and washed once with IX PBS, followed by centrifugation at 400 xg for 10 minutes. Next, the supernatant was removed and the cell pellet was resuspended in 1 mL of IX PBS. After that, the fluorescence signal of DCFH-DA was detected by a flow cytometer (Beckman) with excitation wavelength (450~490nm) and emission wavelength (emission wavelength) 510~550nm, thereby calculating the damage caused by ROS. percentage of cells. Statistically significant differences between groups were determined by Student's t-test. The results of this example are shown in FIG. 3 .

Figure 3 is a data graph of the efficacy of the present rose plant extract in reducing the generation of free radicals in cells caused by hydrogen peroxide. As can be seen from Figure 3, compared with the control group, the percentage of cells damaged by ROS (i.e., cells with high oxidative stress) measured in the hydrogen peroxide group was significantly increased, which indicated that hydrogen peroxide would produce skin fibroblasts. Compared with the hydrogen peroxide group, the percentage of cells damaged by ROS in the experimental group was significantly reduced (reduced ROS expression by at least 97%). The results of this example show that the rose plant extract of the present invention has the effect of reducing the generation of free radicals in cells caused by hydrogen peroxide.

Example 5. Evaluation of the efficacy of rose plant extracts in inhibiting blue light-induced melanin production

First, the mouse skin melanoma cell line B16F10 (corresponding to ATCC CRL-6475) was cultured in Dulbecco's Modified Eagle's Medium (DMEM) supplemented with 1% penicillin/streptomycin (Gibco ) and 10% FBS (Gibco)). 3 mL of medium was added to each well of a 6-well plate to give ^1.5 x 105 B16F10 cells per well. After 24 hours of incubation at 37°C, the medium was removed.

After that, the B16F10 cells were divided into three groups, including the experimental group, the blue light group, and the control group. 0.5% (v/v) rose plant extract was added to the cells of the experimental group, cultured at 37°C for 1 hour, and irradiated with blue light for 3 hours. The cells in the blue light group were irradiated with blue light for 3 hours, while the cells in the control group were not added with rose plant extract and were not irradiated with blue light.

After each group of cell cultures was incubated at 37°C for 48 hours, the medium was removed and washed twice with IxPBS (Gibco). After that, trypsin was added to treat the cells for 3 minutes and the suspended cells were collected in a centrifuge tube with a volume of 15 mL, followed by spinning at 400×g/5 minutes to pellet the cells. After rinsing twice with 1xPBS, the cell pellet was resuspended in 200 [mu]L of 1XPBS. Next, the cell solution was placed in liquid nitrogen for 10 minutes, and then stood at room temperature for 30 minutes to thaw. After complete thawing, spin at 12,000 xg for 30 minutes, then remove the supernatant and add 120 [mu]L of IN _NaOH (as ddH2O). After mixing well, it was left to stand in a 60°C dry bath for 1 hour. After that, a volume of 100 μL was taken into a 96-well culture dish and the absorbance value (OD ₄₅₀ ) of each well was read with an ELISA reader at a wavelength of 450 nm.

The melanin content (%) is calculated by substituting the measured absorbance value (OD ₄₅₀ ) into the following formula (1):

Melanin content (%)=(OD ₄₅₀ absorbance value measured in each group/OD ₄₅₀ absorbance value measured in control group)×100% (1)

Statistically significant differences between groups were determined by Student's t-test. The results of this example are shown in FIG. 4 .

Figure 4 is a data graph showing the efficacy of the rose plant extract of the present invention in inhibiting blue light-induced melanin production. As can be seen from Figure 4, compared with the control group, the melanin content of the blue light group was significantly increased, which means that blue light will produce a large amount of melanin in cells; compared with the blue light group, the melanin content of the experimental group was significantly reduced. (Compared with the blue light group, the melanin content of the experimental group was reduced by at least 38.8%). The results of this example show that the rose flower extract of the present invention has the effect of inhibiting the production of melanin caused by blue light.

In summary, the present rose plant extract can reduce the free radical generation (eg, caused by blue light) in cells (eg, skin fibroblasts) by increasing the mitochondrial activity of cells (eg, skin fibroblasts). And inhibit melanin production (for example, caused by 3C blue light products such as mobile phones and tablet computers), to protect skin cells, strong anti-oxidation and anti-aging effects, thereby maintaining skin youthful vitality.

The above description is exemplary only, not limiting. Any equivalent modifications or changes that do not depart from the spirit and scope of the present invention shall be included in the appended patent application scope.

Claims (6)

Use of a rose ( ROSA ) plant extract for preparing a composition for enhancing mitochondrial activity in human skin cells, wherein the rose plant extract is obtained by extracting a rose plant with a solvent, the solvent is water, wherein the rose plant extract is a rose ( Rosa rugosa ) flower extract, and wherein the extraction is performed at a temperature between 65 and 85°C. The use according to claim 1, wherein the cell is a dermal fibroblast. The use as described in claim 1, wherein the effective concentration of the rose flower extract is at least 0.25% (v/v). The use as described in item 1 of the patent application scope, wherein the volume ratio of the solvent to the rose is between 5-20:1-5. The use as described in item 1 of the scope of the application, wherein the composition is a medicine, a food product or a skin care product. The use as described in item 5 of the claimed scope, wherein the medicinal product comprises a pharmaceutically acceptable carrier.

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