Patent Description:
A material implanted into living tissues should be non-toxic and, after the desired function has been achieved, degraded by metabolic activities and cleared out of the body. Recently, implantation of specific substances into the skin tissue for cosmetic purposes is increasing rapidly. Hyaluronic acid, which is commonly used for this purpose, is known to be metabolized and cleared out of the body within a few days after insertion or implantation regardless of the concentration or the type of the composition. To maintain the desired cosmetic effect for a long period of time, it is necessary to overcome the short period of metabolism of the hyaluronic acid in the human body. Therefore, needs on the development of a new filler material that can be maintained in the human body for months or longer are increasing.

<CIT> discloses tissue augmentation compositions with adjustable persistence properties. Components such as enzyme inhibitors which would tend to prevent degradation of relevant constituents, such as for example hyaluronidase inhibitors, may be included in the compositions. <CIT> discloses soft tissue filler compositions comprising hyaluronic acid in combination with an autologous dermis-derived cell culture material providing a long-term maintenance.

The inventors of the present disclosure have confirmed that, when oral administration of collagen peptide is accompanied by injection of a hyaluronic acid filler, the collagen peptide can lengthen the in-vivo duration of the hyaluronic acid filler injected into the skin and have completed the present disclosure.

The present disclosure is directed to providing a composition that can maintain the effect of a hyaluronic acid filler for a long period of time after injection thereof.

The present invention is defined in the appended claims and relates to the use of a composition for maintaining the effect of a hyaluronic acid filler which contains collagen as an active ingredient.

Since a composition according to the present disclosure can promote the synthesis of hyaluronic acid and inhibit the activity of hyaluronidase, it is useful in maintaining the effect of a hyaluronic acid filler for a longer period of time. As a result, the effect of the filler can be maintained even with a smaller number of filler injections and, thus, skin irritation can be reduced.

Collagen is one of scleroproteins and contains a small amount of sugar. It is the main component of connective tissues and makes up about <NUM> wt% of the whole-body protein content in mammals. Collagen is present in the form of fibrils in the dermis, cartilage, etc. of animals and multiple collagen fibrils form into collagen fibers. The basic unit constituting the fiber is tropocollagen with a molecular weight of about <NUM>,<NUM>, which is approximately <NUM> long and <NUM> in diameter.

The composition according to the present disclosure may maintain a hyaluronic acid filler more naturally for a longer period of time after being injected into the skin.

The collagen peptide of the present disclosure may promote the synthesis of hyaluronic acid and may enhance the activity or quantity of enzymes or proteins involved in the synthesis of hyaluronic acid.

In the present disclosure, the 'hyaluronic acid filler' may mean a filler containing hyaluronic acid (HA) as a main ingredient. The hyaluronic acid is one of complex polysaccharides composed of amino acids and uronic acids, and is a polymer compound consisting of N-acetylglucosamine and glucuronic acid. The filler may mean a substitute material directly injected or inserted into the skin to fill wrinkles, depressed scars, etc. The filler may be used for various purposes without limitation, including alleviation of skin wrinkles, replenishment of moisture, or the like. The filler may be injected into the any part of the body, including, specifically, face, neck, etc..

In the present disclosure, 'to maintain the effect of a filler' may mean that the injected filler is degraded more slowly or that the content of hyaluronic acid which is the main ingredient of the filler is increased by promoting the synthesis of hyaluronic acid in vivo.

The composition of the present disclosure is useful since it can maintain the effect of a filler, including but not being limited to alleviation of skin wrinkles, replenishment of moisture, etc., for a long period of time. Specifically, it is advantageous in terms of skin irritation and economy since the filler injection period can be increased.

The composition according to the present disclosure may promote the synthesis of hyaluronic acid and may also promote the activity or increase the quantity of enzymes, proteins, etc. involved in the synthesis of hyaluronic acid.

In the composition according to the present disclosure, the collagen includes collagen peptide.

In the present disclosure, the collagen peptide is not particularly limited as long as it is a peptide having a molecular weight of <NUM>-<NUM>,<NUM> Da wherein amino acids are linked by peptide bonding. Specifically, the collagen peptide may include a Gly-X-Y collagen tripeptide, wherein X and Y may be any naturally occurring amino acids. The X and the Y may be the same or different amino acids.

The composition according to the present disclosure may contain <NUM>-<NUM> wt% of collagen peptide based on the total weight of the composition. Within this range, the composition may maintain the effect of the hyaluronic acid filler for a long period of time by promoting the synthesis of hyaluronic acid. In this aspect, the composition according to the present disclosure may contain the collagen peptide in an amount of <NUM>-<NUM> wt% based on the total weight of the composition, <NUM>-<NUM> wt% based on the total weight of the composition, <NUM>-<NUM> wt% based on the total weight of the composition, <NUM>-<NUM> wt% based on the total weight of the composition or <NUM>-<NUM> wt% based on the total weight of the composition.

In the composition according to the present disclosure, the collagen tripeptide may contain a Gly-X-Y collagen tripeptide.

In the composition according to the present disclosure, the X and the Y may be the same or different amino acids and the amino acid may be selected from a group consisting of glycine (Gly), alanine (Ala), valine (Val), leucine (Leu), isoleucine (Ile), threonine (Thr), serine (Ser), cysteine (Cys), methionine (Met), aspartic acid (Asp), asparagine (Asn), glutamic acid (Glu), glutamine (Gln), lysine (Lys), arginine (Arg), histidine (His), phenylalanine (Phe), tyrosine (Tyr), tryptophan (Trp) and proline (Pro).

Specifically, the collagen tripeptide may include glycine-proline-hydroxyproline.

The composition according to the present disclosure may contain the collagen or the collagen peptide in an amount of <NUM>-<NUM> wt% based on the total weight of the composition. When the content of the collagen or the collagen peptide is less than <NUM> wt%, it is difficult to achieve the desired effect. And, when it exceeds <NUM> wt%, formulation may be difficult. In this aspect, the composition of the present disclosure may contain <NUM>-<NUM> wt%, <NUM>-<NUM> wt%, <NUM>-<NUM> wt%, <NUM>-<NUM> wt%, <NUM>-<NUM> wt%, <NUM>-<NUM> wt%, <NUM>-<NUM> wt%, <NUM>-<NUM> wt%, <NUM>-<NUM> wt% or <NUM>-<NUM> wt% of the collagen or the collagen peptide based on the total weight of the composition.

In the composition according to the present disclosure, the collagen or the collagen peptide may inhibit the activity of hyaluronidase. The collagen or the collagen peptide may inhibit the degradation of hyaluronic acid by inhibiting the activity of hyaluronidase or reducing its quantity. Accordingly, the synthesis of hyaluronic acid may be promoted and the effect of the filler may be maintained for a long period of time.

The composition according to the present disclosure may be a pharmaceutical composition.

The pharmaceutical composition according to the present disclosure may be prepared into a solid, semisolid or liquid formulation for oral administration by adding a commonly used inorganic or organic carrier to the active ingredient.

The formulation for oral administration may be a tablet, a pill, a granule, a capsule, a powder, a fine granule, a dust, an emulsion, a syrup, a pellet, etc. The active ingredient of the present disclosure may be easily prepared into the desired formulation according to a commonly employed method by using a surfactant, an excipient, a colorant, a flavoring agent, a preservative, a stabilizer, a buffer, a suspending agent or other commonly used adjuvants.

The pharmaceutical composition according to the present disclosure may be administered orally.

The dosage of the active ingredient will vary depending on the age, gender and body weight of a subject, particular disease or pathological condition to be treated and severity thereof, administration route or the discretion of a diagnoser. Determination of the dosage considering these factors is in the level of those skilled in the art. A general dosage is <NUM>-<NUM>,<NUM>/kg/day, more specifically <NUM>-<NUM>,<NUM>/kg/day.

The composition according to the present disclosure may also be a food composition.

In an aspect of the present disclosure, the food composition may be formulated into many forms including but not limited to a powder, a granule, a tablet, a capsule, a drink, etc..

If necessary, the food composition may contain one or more of the additives described below. The additive may be a concentrated fruit juice or a fruit juice powder of grape, apple, orange, lemon, pineapple, banana, pear, etc., a water-soluble or oil-soluble vitamin such as retinol palmitate, riboflavin, pyridoxine, cyanocobalamin, ascorbic acid, nicotinamide, calcium pantothenate, folic acid, biotin, cholecalciferol, choline bitartrate, tocopherol, β-carotene, etc., a flavor such as lemon flavor, orange flavor, strawberry flavor, grape flavor, vanilla essence, etc., an amino acid such as glutamic acid, sodium glutamate, glycine, alanine, aspartic acid, sodium aspartate, inosinic acid, etc., a nucleic acid or salts thereof, a plant fiber such as polydextrose, pectin, xanthan gum, glucomannan, alginic acid, etc., or a mineral such as sodium chloride, magnesium sulfate, potassium chloride, magnesium chloride, magnesium carbonate, calcium chloride, dipotassium phosphate, monosodium phosphate, calcium glycerophosphate, sodium ferrous citrate, ammonium ferric citrate, ferric citrate, manganese sulfate, copper sulfate, sodium iodide, potassium sorbate, zinc, manganese, copper, iodine, cobalt, etc..

The present invention further relates to the use as defined in the appended claims of a kit for injecting a filler, which contains a hyaluronic acid filler composition and the composition according to the present disclosure.

The kit according to the present disclosure may further contain an instruction which instructs that the hyaluronic acid is administered transdermally and the composition according to the present disclosure is administered orally.

The kit of the present disclosure may be administered orally after the injection of a filler so as to prolong the duration of the effect of the filler.

In another aspect, the present disclosure may relate to a use of collagen for preparation of a composition for maintaining the effect of a hyaluronic acid filler. The use concerns non-therapeutic uses.

In the use for preparation of a composition for maintaining the effect of a hyaluronic acid filler according to the present disclosure, the collagen includes collagen peptide.

In the use for preparation of a composition for maintaining the effect of a hyaluronic acid filler according to the present disclosure, the composition may contain <NUM>-<NUM> wt% of collagen peptide based on the total weight of the composition. Within this range, the composition may maintain the effect of the hyaluronic acid filler for a long period of time by promoting the synthesis of hyaluronic acid. In this aspect, the composition according to the present disclosure may contain the collagen peptide in an amount of <NUM>-<NUM> wt% based on the total weight of the composition, <NUM>-<NUM> wt% based on the total weight of the composition, <NUM>-<NUM> wt% based on the total weight of the composition, <NUM>-<NUM> wt% based on the total weight of the composition or <NUM>-<NUM> wt% based on the total weight of the composition.

In the use for preparation of a composition for maintaining the effect of a hyaluronic acid filler according to the present disclosure, the collagen tripeptide may contain a Gly-X-Y collagen tripeptide.

In the use for preparation of a composition for maintaining the effect of a hyaluronic acid filler according to the present disclosure, the X and the Y may be the same or different amino acids and the amino acid may be selected from a group consisting of glycine (Gly), alanine (Ala), valine (Val), leucine (Leu), isoleucine (Ile), threonine (Thr), serine (Ser), cysteine (Cys), methionine (Met), aspartic acid (Asp), asparagine (Asn), glutamic acid (Glu), glutamine (Gln), lysine (Lys), arginine (Arg), histidine (His), phenylalanine (Phe), tyrosine (Tyr), tryptophan (Trp) and proline (Pro).

In the use for preparation of a composition for maintaining the effect of a hyaluronic acid filler according to the present disclosure, the X may be proline and the Y may be hydroxyproline.

In the use for preparation of a composition for maintaining the effect of a hyaluronic acid filler according to the present disclosure, the composition according to the present disclosure may contain the collagen or the collagen peptide in an amount of <NUM>-<NUM> wt% based on the total weight of the composition. When the content of the collagen or the collagen peptide is less than <NUM> wt%, it is difficult to achieve the desired effect. And, when it exceeds <NUM> wt%, formulation may be difficult. In this aspect, the composition of the present disclosure may contain <NUM>-<NUM> wt%, <NUM>-<NUM> wt%, <NUM>-<NUM> wt%, <NUM>-<NUM> wt%, <NUM>-<NUM> wt%, <NUM>-<NUM> wt%, <NUM>-<NUM> wt%, <NUM>-<NUM> wt%, <NUM>-<NUM> wt% or <NUM>-<NUM> wt% of the collagen or the collagen peptide based on the total weight of the composition.

In the use for preparation of a composition for maintaining the effect of a hyaluronic acid filler according to the present disclosure, the collagen or the collagen peptide may inhibit the activity of hyaluronidase.

In the use for preparation of a composition for maintaining the effect of a hyaluronic acid filler according to the present disclosure, the composition according to the present disclosure may be a pharmaceutical composition or a health food composition.

In another aspect, the present disclosure may relate to a method for maintaining the effect of a hyaluronic acid filler in a subject by administering a composition containing collagen to the subject. The subject is one who has been administered with the hyaluronic acid filler prior to the administration of the composition.

In the method for maintaining the effect of a hyaluronic acid filler according to the present disclosure, the collagen includes collagen peptide.

In the method for maintaining the effect of a hyaluronic acid filler according to the present disclosure, the composition may contain <NUM>-<NUM> wt% of collagen peptide based on the total weight of the composition. Within this range, the composition may maintain the effect of the hyaluronic acid filler for a long period of time by promoting the synthesis of hyaluronic acid. In this aspect, the composition according to the present disclosure may contain the collagen peptide in an amount of <NUM>-<NUM> wt% based on the total weight of the composition, <NUM>-<NUM> wt% based on the total weight of the composition, <NUM>-<NUM> wt% based on the total weight of the composition, <NUM>-<NUM> wt% based on the total weight of the composition or <NUM>-<NUM> wt% based on the total weight of the composition.

In the method for maintaining the effect of a hyaluronic acid filler according to the present disclosure, the collagen tripeptide may contain a Gly-X-Y collagen tripeptide.

In the method for maintaining the effect of a hyaluronic acid filler according to the present disclosure, the X and the Y may be the same or different amino acids and the amino acid may be selected from a group consisting of glycine (Gly), alanine (Ala), valine (Val), leucine (Leu), isoleucine (Ile), threonine (Thr), serine (Ser), cysteine (Cys), methionine (Met), aspartic acid (Asp), asparagine (Asn), glutamic acid (Glu), glutamine (Gln), lysine (Lys), arginine (Arg), histidine (His), phenylalanine (Phe), tyrosine (Tyr), tryptophan (Trp) and proline (Pro).

In the method for maintaining the effect of a hyaluronic acid filler according to the present disclosure, the X may be proline and the Y may be hydroxyproline.

In the method for maintaining the effect of a hyaluronic acid filler according to the present disclosure, the composition according to the present disclosure may contain the collagen or the collagen peptide in an amount of <NUM>-<NUM> wt% based on the total weight of the composition. When the content of the collagen or the collagen peptide is less than <NUM> wt%, it is difficult to achieve the desired effect. And, when it exceeds <NUM> wt%, formulation may be difficult. In this aspect, the composition of the present disclosure may contain <NUM>-<NUM> wt%, <NUM>-<NUM> wt%, <NUM>-<NUM> wt%, <NUM>-<NUM> wt%, <NUM>-<NUM> wt%, <NUM>-<NUM> wt%, <NUM>-<NUM> wt%, <NUM>-<NUM> wt%, <NUM>-<NUM> wt% or <NUM>-<NUM> wt% of the collagen or the collagen peptide based on the total weight of the composition.

In the method for maintaining the effect of a hyaluronic acid filler according to the present disclosure, the collagen or the collagen peptide may inhibit the activity of hyaluronidase.

In the method for maintaining the effect of a hyaluronic acid filler according to the present disclosure, the composition according to the present disclosure may be a pharmaceutical composition or a health food composition.

Hereinafter, formulation examples of the composition of the present disclosure will be described. However, the following formulation examples are for illustrative purposes only and various formulations of a pharmaceutical composition and a health food composition are possible.

According to a commonly employed method, <NUM> of the collagen peptide of Preparation Example <NUM>, <NUM> of palm oil, <NUM> of hydrogenated palm oil, <NUM> of yellow beeswax and <NUM> of lecithin were mixed. Then, a soft capsule was prepared by filling <NUM> of the mixture per capsule.

<NUM> of the collagen peptide of Preparation Example <NUM>, <NUM> of glucose, <NUM> of red ginseng extract, <NUM> of starch and <NUM> of magnesium stearate were mixed. After forming granules by adding <NUM> of <NUM>% ethanol, the granules were dried at <NUM> and then compressed into a tablet.

<NUM> of the collagen peptide of Preparation Example <NUM>, <NUM> of glucose, <NUM> of red ginseng extract and <NUM> of starch were mixed. After forming granules by adding <NUM> of <NUM>% ethanol, the granules were dried at <NUM> and then filled in a pouch. The final weight of the content was <NUM>.

<NUM> of the collagen peptide of Preparation Example <NUM>, <NUM> of glucose, <NUM> of red ginseng extract, <NUM> of citric acid and <NUM> of purified water were mixed and filled in a bottle. The final volume of the content was <NUM>.

Hereinafter, the present disclosure will be described in detail through examples. The following examples are for illustrative purposes only.

Collagen peptide used as the active ingredient in the present disclosure, which was purchased from Jellice (HATC, Jellice Co. , Japan), contains <NUM>% or more of a tripeptide such as glycine-proline-hydroxyproline.

Human-derived HaCaT keratinocytes (Dr N. Fusenig, Deutsches Krebsforschungszentrum, Heidelberg, Germany) were cultured in a DMEM medium containing <NUM>% FBS under the condition of <NUM> and <NUM>% CO<NUM>. The cells were cultured on a <NUM>-well plate. Upon reaching <NUM>% confluence, the medium was replaced with an FBS-free medium and the cells were cultured for <NUM> hours. Subsequently, the cells were treated with the collagen peptide of Preparation Example <NUM>, diluted <NUM>-fold in PBS with a final concentration of <NUM> ppm or <NUM> ppm, and then cultured for <NUM> hours. <NUM> hours later, the medium was recovered and used for hyaluronic acid (HR) assay. The remaining cells were subjected to quantitative cell viability (CCK8) assay.

The CCK8 assay (Dojindo) and the hyaluronic acid assay (Echelon) were conducted according to the protocols of the kit manufacturers. The quantitated hyaluronic acid value was corrected by dividing with the cell viability value.

As a result, the collagen peptide-treated group showed <NUM>-<NUM>% increased hyaluronic acid synthesis as compared to the untreated control group, as seen from <FIG>. The promotion of the hyaluronic acid synthesis was proportional to the concentration of the collagen peptide. Accordingly, it was confirmed that the collagen peptide can promote the synthesis of hyaluronic acid in skin cells.

The effect of inhibiting the activity of hyaluronidase was investigated to confirm whether the hydrolysis of hyaluronic acid which is the main ingredient of the filler is inhibited.

<NUM>µL of the collagen peptide was added to <NUM>µL of a <NUM> hyaluronidase solution (<NUM>,<NUM> units/mL), to a final concentration of <NUM>, <NUM>, <NUM>, <NUM> or <NUM>/mL. After mixing with <NUM>µL of <NUM> CaCl<NUM> to activate the enzyme, incubation was performed in an aqueous solution at <NUM> for <NUM> minutes. For the control group, distilled water was added instead of the collagen peptide and incubation was performed for <NUM> minutes in an aqueous solution. After adding <NUM>µL of a <NUM> hyaluronic acid solution (<NUM>/<NUM>) to the hyaluronidase solution activated with Ca<NUM>+, incubation was performed again in an aqueous solution for <NUM> minutes. After the incubation, <NUM>µL of a <NUM> N NaOH solution and <NUM>µL of <NUM> potassium tetraborate were added to the reaction mixture and then cooled after incubation in a boiling water bath for <NUM> minutes. After adding <NUM> of a dimethylaminobenzaldehyde solution (a mixture of <NUM> of p-dimethylaminobenzaldehyde, <NUM> of <NUM>% acetic acid and <NUM> of <NUM> N HCl) to the cooled reaction mixture and incubating in a water bath at <NUM> for <NUM> minutes, absorbance was measured at <NUM>.

As can be seen from Table <NUM>, the collagen peptide exhibited superior effect of inhibiting the activity of hyaluronidase. The effect was dependent on the concentration of the collagen peptide.

After injecting a commercially available hyaluronic acid filler (Glytone II, AmorePacific, <NUM>/g HA) into the back of hairless mice, the duration of volume in the skin was observed.

<NUM>- to <NUM>-week-old hairless mice were divided into two groups. One group was given a normal diet (control groups) and the other group was orally administered with <NUM>/kg collagen peptide for <NUM> weeks (test groups). The control groups and the test groups were subdivided as described in Table <NUM> by varying the injection volume of the filler.

After filler injection, both the control groups and the test groups were given a normal diet and only the test groups were orally administered with the collagen peptide. Then, the volume change of the control groups and the test groups was observed for <NUM> weeks. As can be seen from <FIG> and <FIG>, the control groups showed faster and consistently decreasing volume of the filler. In contrast, the test groups which were orally administered with the collagen peptide showed superior effect of maintaining the volume as compared to the control groups.

Claim 1:
A non-therapeutic use of collagen peptide for maintaining a filling
effect of a hyaluronic acid filler, wherein the collagen peptide is administered to a subject after the hyaluronic acid filler is administered to the subject,
and wherein the collagen peptide comprises a Gly-X-Y collagen tripeptide, preferably, the X and the Y are the same or different amino acids and the amino acid is selected from a group consisting of glycine (Gly), alanine (Ala), valine (Val), leucine (Leu), isoleucine (Ile), threonine (Thr), serine (Ser), cysteine (Cys), methionine (Met), aspartic acid (Asp), asparagine (Asn), glutamic acid (Glu), glutamine (Gln), lysine (Lys), arginine (Arg), histidine (His), phenylalanine (Phe), tyrosine (Tyr), tryptophan (Trp) and proline (Pro), more preferably, the X is proline and the Y is hydroxyproline.