KR20140114617A - Hydrogel patch containing the curcumin for wound healing and preparation method thereof - Google Patents

Abstract

Provided are a pharmaceutical or cosmetic composition and an application-type gel, which prepare a curcumin-containing gel by adding curcumin to gel formulations for transdermal delivery to check an effect of treating wound, and treat skin wound containing the same. Through in vitro anti-inflammatory experiments and in vivo animal experiments, the present invention can check an increase of reepithelialisation tendency on a wounded area when using the curcumin-containing gel, thus is capable of being used for preparing the application-type gel which promotes wound healing.

Description

[0001] The present invention relates to a coated gel for curative treatment containing curcumin and a preparation method thereof,

The present invention relates to a coated gel for wound healing which comprises gelled curcumin as an active ingredient and a method for producing the same.

Skin is an organ that has the function of protecting the human body and exposed to the dangerous situation when the skin is damaged. Skin externalities such as trauma caused by external forces, skin damage caused by skin diseases, and skin defects such as burns are formed, and the skin tries hard to minimize the signs of rapid tissue regeneration and regenerated tissue. Therefore, prompt treatment is required to restore damaged skin tissue without side effects, and wound treatment with an appropriate wound treatment agent is essential. The development of many medicines has been actively researched for wound healing agents for wound healing. However, many chemical synthesis materials have been developed for tissue repair and wound healing due to oxidative damage, but development of a natural material treatment agent capable of replacing existing chemical synthetic materials has been actively pursued due to the side effect due to resistance, [Kim et al. 2012; Lee et al. 2005].

Centella , a natural herb that can be used without side effects in place of conventional synthetic compounds used to treat skin damage, asiatica ) , aloe vera gel known as a burn and wound treatment agent from ancient times, and natural materials such as propolis that promotes wound healing while reducing side effects Gels, and ointments are commercially available and sold [Blonska et al. 2004; Chiou et al. 2001].

Of curcumin turmeric (curcumin) The main component of the (Curcuma longa) is sseuyimyeo much as a spice, a long time anti-oxidation, anti-inflammatory, various it is known that the physiological activity increases the more the functional value, such as detoxification [Sidhu et al. 1998]. In Korea, Oh et al. Have investigated the composition and efficacy of curcumin in Ulgum, and have reported that it is closely related to the anti-inflammatory effect by increasing the content of flavonoids and polyphenols in Ulguk extract [Oh et al. 2010]. However, curcumin has poor solubility, strong taste, color, and unique fragrance that do not dissolve in water. Therefore, there are many problems that need to be solved in order to use it in foods, medicines and cosmetics.

Previous studies from abroad have reported that nanocapsulated curcumin was prepared in the form of a hydrogel to facilitate transdermal delivery and that collagen in the wound area was rapidly formed [Patel et al. 2011].

It has been reported that a composition containing such curcumine or a pharmaceutically acceptable salt thereof as an active ingredient can be prepared and used for pain relieving and relieving products such as analgesics, functional toothpaste for preventing and improving pain, functional foods and food additives (Korean Patent Laid-Open Publication No. 10-2009-0047637), and development of a flap treatment and treatment drug has been carried out. Based on this, a composition for promoting survival of the flap, a pharmaceutical composition for treating or treating flap, (Korean Patent Laid-Open Publication No. 10-2010-0011645). Korean Patent Laid-Open Publication No. 10-2010-0029672 also discloses that curcumin can be used as a pharmaceutical composition for the treatment or prevention of skin diseases caused after treatment. As a pharmaceutical composition, it was possible to confirm a remarkable therapeutic function for various eye diseases and skin diseases, Dermatitis, it was confirmed that represent a significant therapeutic effect on diseases such as androgenetic alopecia, acne.

However, the use of such curcumin is characterized in that a composition for prevention and treatment of pain is formed by incorporating curcumin as an active ingredient, and a technical structure requiring a carrier is essential.

Published Korean Patent Application No. 10-2009-0047637, May 13, 2009 Korean Patent Publication No. 10-2010-0011645, Feb. 3, 2010 Korean Patent Publication No. 10-2010-0029672, Mar. 17, 2010

J. Kim, Y.M. Kim, D.W. Kim, and K.Y. Lee, J Exp Biomed Sci, 18, 175 (2012) H.S. Lee, J.T. Cheong, H.J. Park, T.K. Shin, J. H. Kim, and J.M. Lee, J Vet Clin, 22 (1), 43 (2005) M. Blonska, J. Bronikowska, G. Pietsz, Z.P. Czuba, S. Scheller, W. Krol, J Ethnopharmacol, 91, 25 (2004) W.F. Chiou, C.J. Chou, C.F. Chen. Life Sci., 69, 625 (2001) G.S. Sidhu, A. K. Singh, D. Thaloor. Wound Repair Regen, 6, 167 (1998) H.I. Oh, H.B. Park, M.S. Ju, S.Y. Jung, M.S. Oh, The Korea Association of Herbology, 25 (1), 83 (2010). N.A. Patel, M. Patel, and R.P. Patel, J Pharm, 01, 15 (2011).

It is an object of the present invention to provide a gel containing curcumin, carbopol 934 polymer, propylene glycol and a process for producing the same, so that curcumin effectively works in wound healing. In addition, it is formulated into a form that can be applied in order to be used as a skin damaging agent, medicine, and cosmetic material.

The present invention relates to a method of treating a skin disease in the form of a coated gel comprising an active ingredient of curcumin, carbopol 934 (polymer) and propylene glycol as an active ingredient, in order to confirm the therapeutic effect of curcumin on a gel formulation for transdermal delivery, A pharmaceutical composition for treatment is provided. Preferably, the skin disease is wound or scar and the composition comprises 0.5 to 3% (w / w) of curcumin, 1 to 5% (w / w) of Carbopol 934 polymer, 1 to 5% To 5% (w / w).

The present invention also provides a cosmetic composition for preventing or improving skin diseases in the form of a coated gel containing curcumin, Carbopol 934 polymer and propylene glycol as effective ingredients, which is effective for re-epithelialisation or skin whitening . Preferably, the skin disease is wound or scar and the composition comprises 0.5 to 3% (w / w) of curcumin, 1 to 5% (w / w) of Carbopol 934 polymer, 1 to 5% To 5% (w / w).

The present invention also provides a method for producing a gel for wound treatment or treatment for transdermal delivery comprising curcumin, Carbopol 934 polymer, and propylene glycol as an active ingredient. Preferably, the coated gel comprises 0.5 to 3% (w / w) of curcumin, 1 to 5% (w / w) of Carbopol 934 polymer, and 1 to 5% (w / w) of propylene glycol.

The coated gel containing curcumin, carbopol 934 and propylene glycol as an active ingredient according to the present invention can be used as a pharmaceutical composition or a cosmetic composition and has an effect of effectively treating or improving skin diseases such as wound and scarring .

Fig. 1 shows the morphology of the curcumin-containing coated gel prepared according to Example 1. Fig.
Fig. 2 shows the cytotoxicity values at each concentration of curcumin gel using RAW 264.7 cells, and showed cell viability of 80% or more at less than 100 ppm of curcumin gel.
FIG. 3 shows the anti-inflammatory effect of curcumin gel in RAW 264.7 cells induced by oxidative stress with lipopolysaccharide, and curcumin gel showed anti-inflammatory effect at 25, 50 and 100 ppm.
FIG. 4 shows the results of wound healing rates obtained by visually and histologically evaluating changes in dorsal skin tissue of wound-induced SD rats in order to evaluate the effect of curcumin gel treatment during wound healing process.

Hereinafter, the present invention will be described in more detail with reference to specific examples. It should be understood, however, that the present invention is not limited by the following embodiments, and that various changes and modifications can be made within the spirit and scope of the present invention.

Unless otherwise defined in the technical and scientific terms used herein, unless otherwise defined, it is understood by those of ordinary skill in the art to which this invention belongs. Repeated descriptions of the same technical constitution and operation as those of the conventional art will be omitted.

Measurement of physiological activity of gelled curcumin and in vitro exam

 1. Physiological activity quantification of curcumin

Total polyphenol content was prepared at a fixed concentration of 1.0 mg / mL. 1.0 mL of Folin-Ciocalteu phenol reagent reagent was added to 3.0 mL of the diluted sample solution, 1.0 mL of saturated solution Na ₂ CO ₃ was added, and the mixture was stirred at room temperature for 1 hour After incubation, the absorbance was measured at 700 nm, and then the absorbance values of the standard curves were compared with the standards tannic acid and gallic acid to calculate the polyphenol content.

The total flavonoid content was determined by adding 1.0 mL of the sample solution dissolved in distilled water, 10.0 mL of diethylene glycol, and 1.0 mL of NaOH. The mixture was well mixed and reacted at room temperature for 1 hour and then absorbed at 510 nm. The calibration curves were calculated using quercetin.

In addition, the electron donating ability for measuring the antioxidative effect was measured. 1.0 mL of 0.2 mM DPPH was added to 2.0 mL of each sample solution. After stirring, the solution was allowed to stand in a dark room for 30 minutes and absorbance was measured at 517 nm. The electron donating ability was expressed by the absorbance reduction rate of the sample solution addition group and the no addition group.

Table 1 below shows the polyphenol content, flavonoid content and electron donating ability of curcumin used in the experiment.

The polyphenols, flavonoid content and electron donating ability of curcumin
Phenolic content (mg / g) Flavonoid content
(mg / g) Electron donating
ablilty IC ₅₀ (ppm) Tannic acid Gallic acid Curcumin 330 ± 0.024 450 ± 0.018 370 ± 0.004 12.5

All values are mean SD of triplicate determinations.

 2. Preparation of curcum-added gel

The manufacture of curcumin gels The existing research method was modified to suit the purpose. The gel to which curcumin is added is characterized by containing curcumin, polymer carbopol 934 and propylene glycol, preferably 0.5 to 3% of curcumin having the properties shown in Table 1 w / w), Carbopol 934 1 to 5% (w / w), and propylene glycol 1 to 5% (w / w).

In this Example, a gel formulation having a composition ratio shown in [Table 2] was prepared and used in future experiments. BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 shows the form of the prepared curcumin containing coated gel.

The composition ratio of the curcumin-containing coated gel prepared in this example Ingredients Formulations Curcumin (%, w / w) One Carbopol 934 (%, w / w) 2 Propylene glycol 2 mL Ethanol 5 mL Triethanolamine qs ^* to neutralize
the gel base Water qs ^*

(Total volume: 100 mL)

* qs: (quantum sales, quantum sufficit)

 3. Cytotoxicity measurement of curcumin gel

Cell viability was measured by Lee et al. [Lee et al., Eur J Pharmacol, 406, p. 301 (2000)). The eluate was prepared by adding the curcumin-added gel to the cell culture medium (DMEM, Dulbecco's modified Eagle's medium, Gibco) at a concentration of 6.25, 12.5, 25, 50, 100 ppm and culturing at 37 ° C for more than 24 hours.

In MTT assay, the MTT (3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyl tetrazolium bromide) reagent is absorbed into the cells and forms formazan by succinate dehydrogenase of mitochondria. 96 RAW 264.7 per each well in the well plate cell 5 × 10 ⁴ cells / well, and the frequency divider 37 ℃ for 24 h gave after transfer to a DMEM culture solution containing the curcumin gel prepared at various concentrations after 24 hours, 5% CO ₂ Lt; / RTI > 20 μL of MTT solution per well was added and incubated at 37 ° C in 5% CO ₂ After incubation for 4 hours, the culture medium was discarded and 100 μL of DMSO was added to dissolve formazan, and then measured at 570 nm using ELISA (ELX 808, Biotek Instruments, Vermont, USA).

Fig. 2 shows the cytotoxicity values at each concentration of curcumin gel using RAW 264.7 cells, and showed cell viability of 80% or more at less than 100 ppm of curcumin gel.

 4. Measurement of nitric oxide production

To determine the amount of nitric oxide (NO) production, the concentration of nitrite in the culture was measured. RAW 264.7 cells were plated at 2.5 × 10 ⁵ cells / well in 48 well plates and cultured for 12 hours. Various concentrations of curcumin gel were treated with Lipopolysaccharide (LPS) (100 ng / ml) for 24 hours, and the amount of NO contained in the culture medium was measured using a Griess Reagent System. Each experiment was independently repeated three times to obtain an average value.

FIG. 3 shows the anti-inflammatory effect of curcumin gel in RAW 264.7 cells induced by oxidative stress with lipid polysaccharide, and the curcumin gel showed anti-inflammatory effect at 25, 50 and 100 ppm.

Effect of curcumin gel on wound healing after wound induction in vivo exam

 1. Experimental animals

Six-week-old Sprague-Dawley (SD) male rats weighing 180 ~ 220 g were purchased from Orient Bio Inc. and maintained at 25 ± 1 ° C and 50 ± 10% Automatically adjusted and stored. In addition, feed and water were allowed to be ingested freely, and all experimental animals were adapted to the animal room environment for one week and then used in the experiment. Animal experiments were approved for conformity with the Ethical Commission of Animal Experiments at the Yonbong Campus of Chonnam National University (CNU IACUC YB-2012-36).

 2. Wound induction and treatment

The subject SD rats were fasted for 24 hours before anesthesia and water was supplied. In all experimental groups, zoletil and rumun were mixed at a ratio of 1: 2 before the wound-induced induction, and were anesthetized with an injection of 0.1 cc / kg using an intramuscular injection. After disinfection, 10% povidine-iodine and 70% ethanol were applied to the dorsal area, and all-thickness defect wounds including dermal layer were formed with a diameter of 2 cm x 2 cm using disposable surgical scissors.

To measure the wound healing effects of curcumin-containing gels, a control gel containing no curcumin and an experimental group treated with curcumin-containing gel were applied to each of the four test tubes four times, about 3 mL each, according to the elapsed days. Afterwards, sterilized gauze (Tegaderm; 3M Health Care, USA) was covered to prevent loss of gel, and the elastic band (Coban 3M HealthCare, USA) was wound thereon to prevent movement.

 3. Observation of gross wound changes

In order to observe the progress of wound healing, we observed the change of the wound with a digital camera (CAMEDIATM, Olympus Co., Tokyo, Japan) . In addition, the area of the wound was measured using digital calliper (Mitutoyo Co., Kawasaki, Japan) to represent the objective index. The wound healing rate was calculated by the method proposed in the previous study [Chang et al., J Vet Clin., 22 (1) p. 43 (2005).

Wound healing rate (%) = 100 x (Wo - Ui) / Wo

Treatment share per period (%) = 100 x (Ub - Ui) / Wo

Wo: Area above the window immediately after wounding

Ui: Measurement window top area

Ub: previous measurement day window top area

 4. Histological examination

After 20 days from induction of wounding, the experimental animals were treated with ethyl ether for histological examination according to the method of overdose of psychotropic drugs specified in the standards for laboratory animal control guidelines of the Ministry of Health and Welfare notification No. 88-9. It was euthanized. The tissues containing the whole round wound were collected and fixed in 10% neutral formalin solution for 24 hours. The sections passing through the center of the wound were dehydrated and embedded in paraffin block. Tissue was cut using a tissue sectioner, then stained with polylysine-coated slides and subjected to paraffin removal and functional procedures followed by hematoxylin-eosin (H & E) staining.

 5. Treatment results using curcumin gel

FIG. 4A is a photograph of the wound of the same subject selected from the control group of the wound-induced group and the experimental group treated with the curcumin-containing gel at 5-day intervals from the wound-induced day to the 20-day post wound- In order to evaluate the efficacy of curcumin gel treatment during wound healing, wounds on the back skin of SD rats were induced and the changes of skin texture during this process were observed. It was observed that the curative treatment process became active due to the application of curcumin gel to the wound. In the group treated with curcumin gel, epidermalization and contraction of incisions were faster, and the wound area was significantly reduced compared with the control group, which was not supplemented with curcumin.

Figure 4B shows the control and experimental groups after H & E staining of SD rats skin. In the experimental group, the surface layer of the epithelial layer was uniform and the surface layer was formed more rapidly than the control group, and the degree of inflammation was smaller than that of the control group.

The change in wound healing rate was measured as shown in Fig. 4C. The wound healing rate was 26.28 ± 1.03% at the 5th day after wounding, 13.57 ± 0.64% at the control group, 46.67 ± 1.46% at the 10th day, 38.21 ± 0.66% at the control group, 81.4 ± 0.47 at the 15th day, 70.21 ± 0.91% At the first day, the experimental group was 90.19 ± 0.35 and the control group was 81.4 ± 1.24%. The wound healing rate from the fifth day after wounding to the end of the experiment was significantly higher in the experimental group than in the control group (p <0.05). In a previous study by Lee et al., It was found that Opuntia ficusindica ) The extract-treated study showed that the wound contraction reached 50% on day 8 [Lee et al., J Vet Clin., 22 (1) p. 43 (2005). The curcumin-containing gel showed 46.67 +/- 1.46% wound constriction at 10 days.

In addition, the change in treatment share per period was measured as shown in FIG. 4D. Between 0 and 5 days, the experimental group was 26.28 ± 1.03%, the control group was 13.57 ± 0.64%, and between 5 and 10 days, the experimental group was 29.66 ± 0.95 and the control group was 17.35 ± 1.04. On the day of wound invasion, blood and exudates and inflammation were secreted in all wounded groups. These acute inflammatory changes persisted until 10 days after wounding. In particular, after 5 days of wounding, foreign bodies were frequently adhered to the wounds due to the large amount of inflammatory exudates in the wound of the control group. On the other hand, in the group treated with curcumin - containing gel, the inflammatory reaction was suppressed and edema and exudate secretion were reduced. After 13 days of wound onset, no further effusion was evident, and the wound was recovering as the wound area decreased.

The epithelium of the experimental group was rapidly matured and the keratinization layer was observed. As reported by Sidhu et al., The curcumin-containing gel exhibited fibronectin and factor-β1 And induce the re-epithelialization by reducing cell death, thereby enhancing wound healing power. These results indicate that the curcumin-containing gel has excellent therapeutic effects on the wound.

Claims (8)

A pharmaceutical composition for the treatment or treatment of a skin disease in the form of a coated gel containing curcumin, a carbopol 934 polymer, and propylene glycol as an active ingredient. The composition of claim 1, wherein the skin disease is wound or scar. The composition of claim 1 or 2, wherein the composition comprises 0.5 to 3% (w / w) of curcumin, 1 to 5% (w / w) of Carbopol 934 polymer, 1 to 5% (w / w) &Lt; / RTI &gt; A cosmetic composition for preventing or improving a skin disease in the form of a coated gel containing curcumin, Carbopol 934 polymer, and propylene glycol as effective ingredients, which is effective for re-epithelialisation or skin whitening. 5. The composition of claim 4, wherein the skin disease is wound or scar. 6. The composition of claim 4 or 5, wherein the composition comprises 0.5 to 3% (w / w) of curcumin, 1 to 5% (w / w) of Carbopol 934 polymer, 1 to 5% (w / w) &Lt; / RTI &gt; A method for the preparation of a coated gel for wound treatment or treatment for transdermal delivery comprising curcumin, Carbopol 934 polymer, and propylene glycol as an active ingredient. The composition of claim 7, wherein the coated gel comprises 0.5 to 3% (w / w) of curcumin, 1 to 5% (w / w) of Carbopol 934 polymer, and 1 to 5% (w / w) &Lt; / RTI &gt;

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