Collagen stabilizer composition

Abstract

The present invention relates to: a stabilizer composition for stabilizing a liquid composition containing collagen, the stabilizer composition containing 2.5 mM to 5 mM of a phosphate buffer and not containing sodium chloride; the liquid composition using same; a method for stabilizing collagen; and a collagen stabilization use of the composition.

Description

Collagen stabilizer composition

[0001] The present invention relates to a stabilizing composition for stabilizing a liquid composition containing collagen, the composition comprising a phosphate buffer of 2.5 mM to 5 mM and not containing sodium chloride; a liquid composition using the same; a method for stabilizing collagen; and a use of the composition for stabilizing collagen.

[0002]

[0003] Collagen is the largest protein in the human body, both quantitatively and functionally. It is the main component of the extracellular matrix, which forms the framework of all cells in the human body, and more specifically, of connective tissue. Human tissues that contain large amounts of collagen include skin, bone tissue such as bone and cartilage, tendons, and ligaments.

[0004] In addition, collagen can be produced artificially and can be used as an ingredient in various fields such as cosmetics for skin improvement such as elasticity enhancement or moisturizing, patches, scaffolds for increasing biocompatibility, implants, capsules, and foods (KR 10-2021-0063600 A).

[0005]

[0006] When collagen is stored as a liquid composition, it may exhibit low stability, such as phase separation or failure to maintain pH under general buffer conditions. Therefore, research on methods for stabilizing liquid compositions containing collagen is necessary.

[0007]

[0008] One object of the present invention is to provide a stabilizer composition comprising a phosphate buffer of 2.5 mM to 5 mM, wherein the stabilizer composition does not contain sodium chloride, and which stabilizes a liquid composition comprising collagen.

[0009] Another object of the present invention is to provide a liquid composition comprising collagen, comprising the stabilizer composition of the present invention.

[0010] Another object of the present invention is to provide a method for stabilizing a liquid composition containing collagen using the stabilizer composition of the present invention.

[0011] Another object of the present invention is to provide a use of the stabilizing composition of the present invention for stabilizing a liquid composition containing collagen.

[0012]

[0013] The composition of the present invention can effectively stabilize collagen for a long period of time.

[0014]

[0015] Figures 1 to 10 are diagrams showing the results of long-term stability phase separation.

[0016] Figures 11 to 14 are diagrams showing the results of long-term stability phase separation according to the sugar alcohol weight ratio.

[0017]

[0018] This is explained in detail as follows. Meanwhile, each description and embodiment disclosed in the present invention can also be applied to each other description and embodiment. That is, all combinations of the various elements disclosed in the present invention fall within the scope of the present invention. Furthermore, the scope of the present invention should not be considered limited by the specific descriptions described below. In addition, numerous papers and patent documents are referenced and cited throughout this specification. The disclosures of the cited papers and patent documents are incorporated into this specification in their entirety by reference to more clearly explain the level of the technical field to which the present invention belongs and the contents of the present invention.

[0019]

[0020] One aspect of the present invention provides a stabilizing composition comprising a phosphate buffer of 2.5 mM to 5 mM, wherein the stabilizing composition does not contain sodium chloride, and wherein the stabilizing composition stabilizes a liquid composition comprising collagen.

[0021] In the present invention, the term "collagen" may refer to a peptide having a molecular weight of 310 Da to 300 kDa, including a tripeptide. In addition, the collagen of the present invention may be naturally derived or artificial collagen (synthetic collagen), but is not limited thereto.

[0022] As an example of implementation, the collagen of the present invention may be various types of artificial collagen, for example, but not limited to, a peptide comprising a tripeptide of glycine-proline-hydroxyproline (Gly-Pro-Hyp) and / or a peptide comprising a tripeptide of proline-hydroxyproline-glycine (Pro-Hyp-Gly).

[0023]

[0024] The term "stabilization" in this application may mean that a liquid composition containing collagen maintains its pH and / or does not undergo phase separation before and after storage. Specifically, the stabilizing agent composition of the present invention may mean that when the liquid composition containing collagen is stored in the stabilizing agent composition of the present invention at a temperature of 25°C and a humidity of 60% for 6 months, the initial pH, which is the pH before storage, is maintained and / or the phases are not separated.

[0025] As an example of implementation, the stabilizer composition of the present invention may prevent phase separation of a liquid composition containing collagen.

[0026] As an example of implementation, the stabilizer composition of the present invention may maintain the pH of a liquid composition comprising collagen.

[0027] Specifically, the stabilizer composition of the present invention may maintain the initial pH when stored for 6 months under conditions of a temperature of 25°C and a humidity of 60%, and in the present invention, pH maintenance may mean that the pH after storage does not exceed ±1 compared to the initial pH.

[0028]

[0029] In one implementation example, the stabilizer composition of the present invention may have a pH of, but is not limited to, 5 to 8. The pH range may be a range in which the stabilizer composition of the present invention can stabilize a liquid composition containing collagen.

[0030] Specifically, the stabilizer composition of the present invention may have a pH of 5 to 7.5, 5 to 7, 5 to 6.5, 5 to 6, 5 to 5.5, 5.5 to 7.5, 5.5 to 7, 5.5 to 6.5, 5.5 to 6, 6 to 7.5, 6 to 7, 6 to 6.5, 6.4 to 7.6, 6.5 to 7.5, or 6.5 to 7, and such pH may be maintained before and after storage, but is not limited thereto.

[0031]

[0032] As an example of one embodiment, the stabilizer composition of the present invention may further comprise a sugar alcohol.

[0033] In addition, in the stabilizer composition of the present invention, the sugar alcohol may be added in place of sodium chloride, and the stabilizer composition containing sodium chloride may undergo phase separation, while the stabilizer composition containing sugar alcohol may not undergo phase separation, but is not limited thereto.

[0034] In the present invention, the sugar alcohol may be mixed with a polyhydric alcohol and may be an alcohol having two or more hydroxyl groups.

[0035] In one embodiment of the above-described embodiment, the sugar alcohol may be glycerol, mannitol, or a combination thereof.

[0036] In one embodiment of the above-described embodiment, glycerol and mannitol in the stabilizer composition may comprise a weight ratio of 1 to 9: 9 to 1, but is not limited thereto. Specifically, glycerol and mannitol may be in a weight ratio of 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1 or 9:1. In one embodiment of the present invention, when collagen was stored for 6 months under general buffer conditions (Tris-buffer 50 mM) or in the presence of sodium chloride, at a temperature of 25°C and a humidity of 60%, phase separation occurred and the liquid composition including collagen was not stabilized, whereas when the present invention was used, it was confirmed that the composition could be stabilized even when stored under the same conditions.

[0037] In particular, when the stabilizing composition of the present invention containing 2.5 mM to 5 mM of phosphate buffer, mannitol, and glycerol but not sodium chloride is used, it was confirmed that the liquid composition containing collagen was stored for 6 months at a temperature of 25°C and a humidity of 60% without phase separation of the liquid composition and the pH was maintained, indicating excellent stability.

[0038] In addition, it was confirmed that when glycerol and mannitol are included in the composition of the present invention, phase separation does not occur, and in particular, when the weight ratio of glycerol and mannitol exceeds the range of 1 to 9: 9 to 1, phase separation occurs and phase stability is not maintained.

[0039]

[0040] Another aspect of the present invention provides a liquid composition comprising collagen and a phosphate buffer of 2.5 mM to 5 mM, which does not contain sodium chloride.

[0041] The above collagen is as described in other aspects, and the liquid composition may be stabilized by including the stabilizer composition of the present invention.

[0042] In one implementation example, the collagen of the present invention may be included in an amount of 1.5 to 3 wt%, but is not limited thereto. Specifically, the collagen included in the composition of the present invention may be included in an amount of about 1.5 to 3 wt%, 1.2 to 3 wt%, 1.0 to 3 wt%, 1.5 to 2.8 wt%, 1.2 to 2.8 wt%, or 1.0 to 2.8 wt%, based on the total weight of the composition, but is not limited thereto.

[0043] The term "about" above encompasses not only the exact number described after the term, but also a range that is or is nearly that number. Whether the number is or is nearly the specific number mentioned can be determined based on the context in which it is presented. For example, the term "about" may refer to a range of -10% to +10% of a given numerical value. As another example, the term "about" may refer to a range of -5% to +5% of a given numerical value. Other examples include, but are not limited to, a range that includes ±0.5, ±0.4, ±0.3, ±0.2, ±0.1, etc.

[0044]

[0045] The liquid composition containing the collagen of the present invention can be manufactured into or included in a tissue repair composition, a composition for biological administration, a pharmaceutical composition, a cosmetic composition, a food composition, and a pharmaceutical composition.

[0046] In addition, another aspect of the present invention provides a tissue repair composition, a composition for bioadministration, a pharmaceutical composition, a cosmetic composition, a food composition, and a pharmaceutical composition, which comprise a liquid composition comprising the collagen of the present invention.

[0047] The liquid composition containing the collagen may be stabilized by including the stabilizer composition of the present invention, as described in other aspects.

[0048]

[0049] The composition of the present invention (e.g., a stabilizer composition, a liquid composition containing collagen, a tissue repair composition, a composition for bioadministration, a quasi-drug composition, a cosmetic composition, a food composition, and a pharmaceutical composition) may further include one or more of a carrier, an excipient, and a diluent.

[0050] Examples of the carrier, excipient, and diluent include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, purified water, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, and mineral oil. For example, when the composition of the present invention is formulated and used, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants may be included in the composition of the present invention, but are not limited thereto.

[0051] Carriers, excipients and diluents can be appropriately selected by those skilled in the art depending on the intended use of the composition.

[0052] Additionally, it is self-evident that the composition of the present invention may further comprise a physiologically active substance and / or a physiologically acceptable carrier.

[0053] The term "physiologically acceptable carrier" in the present invention may include an acceptable carrier in a tissue repair composition, a composition for biological administration, a quasi-drug composition, a cosmetic composition, a food composition, and / or a pharmaceutical composition, and refers to a carrier or diluent that does not stimulate a living organism and does not inhibit the activity and properties of the composition of the present invention. In a composition formulated as a liquid solution, an acceptable carrier may be a sterile and biocompatible one, and depending on the properties of the composition, saline solution, sterile water, albumin injection solution, dextrose solution, maltodextrin solution, ethanol, and one or more of these components may be mixed and used, but is not limited thereto.

[0054]

[0055] In the present invention, the term 'tissue repair' means something that can be used for replacing, repairing, and / or reconstructing human tissues and organs, such as blood vessels, heart, septum, fascia, joints, joint spaces, cartilage, tendons or ligaments, and / or skin.

[0056] The tissue repair composition of the present invention can be used as a tissue repair biomaterial, and the tissue repair biomaterial refers to a bio-derived material used for replacement, repair, and reconstruction of human tissues and organs such as blood vessels, heart, septum, fascia, skin, joints, joint spaces, cartilage, tendons, or ligaments, and is not limited to a cosmetic filler, and can refer to a tissue repair biomaterial specified by the Ministry of Food and Drug Safety.

[0057] The composition for bioadministration of the present invention is not limited in terms of formulation, administration method, etc., as long as it can be administered to a living body.

[0058] The term "administration" in the present invention refers to introducing the composition of the present invention into a subject by an appropriate method, and the administration route may be various, such as application, subcutaneous injection, or dermal injection, as long as it can reach the target tissue. The formulation may be, but is not limited to, a topical preparation, subcutaneous injection (injection), dermal injection (injection), or intramuscular injection (injection). The preferred dosage of the composition of the present invention may vary depending on the condition of the subject.

[0059]

[0060] In the present invention, the term "quasi-drug" means a product that has a milder effect than a pharmaceutical product among products used for the purpose of diagnosing, treating, improving, alleviating, managing or preventing diseases of humans or animals. For example, according to the Pharmaceutical Affairs Act of the Republic of Korea, a quasi-drug is a product excluding products used for the purpose of pharmaceutical products, and includes products used for treating or preventing diseases of humans or animals, products that have a mild effect on the human body or do not act directly, etc.

[0061] As an example of one embodiment, the pharmaceutical composition of the present invention can be manufactured in a form selected from the group consisting of, but not limited to, body cleanser, shampoo, conditioner, foam, soap, mask, ointment, cream, lotion, essence, and spray.

[0062]

[0063] The cosmetic composition of the present invention can be formulated in various forms. In one embodiment, the cosmetic composition of the present invention is a product with specialized functionality, emphasizing physiologically active efficacy and effectiveness, and may be included in cosmetics as defined by the Ministry of Health and Welfare. However, this is not a limitation.

[0064] In one embodiment, the cosmetic composition of the present invention can be prepared in a formulation selected from the group consisting of, but not limited to, a solution, an external ointment, a cream, a foam, a nourishing toner, an emollient toner, a pack, an emollient, a milky lotion, a makeup base, an essence, a soap, a liquid cleanser, a bath agent, a sunscreen cream, a sun oil, a suspension, an emulsion, a paste, a gel, a lotion, a powder, a soap, a surfactant-containing cleanser, an oil, a powder foundation, an emulsion foundation, a wax foundation, a patch, and a spray.

[0065]

[0066] The term "food" of the present invention includes dairy products including meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, ice cream, various soups, beverages, tea, drinks, alcoholic beverages, vitamin complexes, health functional foods, and health foods, and includes all foods in the conventional sense.

[0067] The above-mentioned "health food" can refer to foods that have a more active health maintenance or promotion effect than regular foods, while "health supplement food" refers to foods intended for health supplementation. In some cases, the terms "health functional food," "health food," and "health supplement food" are used interchangeably.

[0068] Additionally, the food composition of the present invention may include additional ingredients commonly used in food compositions to improve efficacy, odor, taste, appearance, etc. (e.g., vitamins, amino acids, minerals, flavoring agents, etc.). The food composition of the present invention may include food additives such as preservatives, colorants, sweeteners, flavoring agents, antioxidants, and flavoring agents. The above additives may be selected depending on the type of food and used in an appropriate amount.

[0069]

[0070] The pharmaceutical composition of the present invention may further comprise a pharmaceutically acceptable carrier. In addition, the pharmaceutically acceptable carrier of the present invention may comprise a non-natural carrier. The pharmaceutical composition of the present invention may be administered in a pharmaceutically effective amount, wherein the term "pharmaceutically effective amount" means an amount sufficient to treat or prevent a disease at a reasonable benefit / risk ratio applicable to medical treatment or prevention, and the effective dosage level may be determined according to the severity of the disease, the activity of the drug, the patient's age, weight, health, sex, the patient's sensitivity to the drug, the time of administration of the composition of the present invention used, the route of administration and the excretion rate, the treatment period, the drug used in combination with or concurrently with the composition of the present invention used, and other factors well known in the medical field. The pharmaceutical composition of the present invention may be administered alone or in combination with a known drug.

[0071]

[0072] Another aspect of the present invention provides a method for stabilizing a liquid composition comprising collagen, the method comprising adding a phosphate buffer of 2.5 mM to 5 mM, wherein sodium chloride is not added.

[0073] The collagen and liquid compositions described above are as described in other aspects.

[0074]

[0075] Another aspect of the present invention provides a use of the stabilizing composition of the present invention for stabilizing a liquid composition comprising collagen.

[0076] The above stabilizer composition, collagen, and stabilization, etc. are as described in other aspects.

[0077]

[0078] The present invention is described in more detail below through examples. However, the following examples are merely preferred embodiments intended to illustrate the present invention and are therefore not intended to limit the scope of the present invention. Furthermore, technical details not described herein can be readily understood and implemented by those skilled in the technical field of the present invention or similar fields.

[0079]

[0080] Example 1. Confirmation of the stability of properties and pH according to various compositions.

[0081] 1-1. Test conditions

[0082] (1) pH conditions: 6.0, 6.5, 7.0, 7.5, 8.0

[0083] (2) Buffer conditions

[0084] - Control: Tris buffer (50 mM) (normal buffer conditions)

[0085] - Experimental group: Phosphate buffer (1, 2.5, 5, 7.5 mM)

[0086] (3) Sugar alcohols (e.g., glycerol, mannitol) or sodium chloride

[0087] (4) Material concentration (main active ingredient): 15 mg / mL to 30 mg / mL In this example, collagen is used as the main active ingredient.

[0088] (5) Stability test conditions

[0089] - Store collagen as an active ingredient for 6 months at a temperature of 25℃ and humidity of 60% under the various conditions mentioned above.

[0090]

[0091] 1-2. Analysis method

[0092] (1) Phase stability: Suitable for maintaining phase (i.e., observing whether phase separation occurs)

[0093] (2) pH stability: Suitable for maintaining pH

[0094] When the stability (pH and long-term stability phase separation) items are met, the stabilizer composition of the present invention can be applied.

[0095]

[0096] 1-3. Results

[0097] The results of the stability analysis according to the above 1-1 and 1-2 are shown in Tables 1 and 2 and Figures 1 to 10. In the case of items in which phase separation was observed, it was confirmed that all components were not mixed but separated and coexisted in two or more liquid phases.

[0098]

[0099] Item Substance Concentration (mg / mL) Buffer Alcohol or Sodium Chloride pH (Initial) pH (After 6 months) Appearance Stability* pH Stability** 13050 mM TrisGlycerolMannitol6.015.97XO26.496.4737.017.0247.497.5057.967.96650mMTrisNaCl7.006.99O7301mMPhosphateGlycerolMannitol6.025.01OX86.465.8296.925.85107.455.89117.975.89X12NaCl6.955.99X13302.5mMPhosphateGlycerolMannitol6.025.99OO146.446.43156.976.9 1167.537.50177.987.96X18NaCl6.955.59XX19305mMPhosphateGlycerolMannitol6.046.00OO206.536.46216.946.95227.557.47237.997.93X24NaCl7.005.75XX25307.5mMPhosphateGlycerolMannitol6.015.97XO266.496.49276.966.91287.487.48297.957.9530NaCl6.936.01XX

[0100] * O: Maintained phase, X: Separated phase

[0101] ** O: pH maintenance, X: pH change

[0102]

[0103] Item Substance Concentration (mg / mL) Buffer Alcohol or Sodium Chloride pH (Initial) pH (6 months) Appearance Stability pH Stability 311550mMTris Glycerol Mannitol 6.00 5.98 XO 326.5 06.49 337.016.99 347.46 7.44 357.97 7.98 3650mMTris NaCl 6.98 6.97 O 37 151mMPhosphate Glycerol Mannitol 6.0 35.00 O X 386.48 5.39 396.99 5.92 407.45 5.95 417.92 5.96 X 42 NaCl 6.90 5.92 X 43 152.5mMPhosphate Glycerol Mannitol 6.0 26. 00OO446.496.45456.976.94467.457.41477.937.90X48NaCl6.955.78XX49155mMPhosphateGlycerolMannitol6.045.99OO506.526.47517.016.97527.527.49537.977.96X54NaCl7.025.92XX55157.5mMPhosphateGlycerolMannitol5.986.01XO566.516.50577.006.99587.517.51597.967.9560NaCl7.026.03XX

[0104] * O: Maintained phase, X: Separated phase

[0105] ** O: pH maintenance, X: pH change

[0106]

[0107] From the above results, it was confirmed that when collagen was stored for 6 months under general buffer conditions (Tris-buffer 50 mM) or in the presence of sodium chloride, at a temperature of 25°C and a humidity of 60%, phase separation occurred and the liquid composition containing collagen could not be stabilized, whereas when the composition of the present invention was used, it could be stabilized even when stored under the same conditions.

[0108]

[0109] Example 2. Confirmation of property stability and pH stability according to sugar alcohol ratio

[0110] Based on the test conditions of Example 1-1 and the analysis method of 1-2, the phase stability and pH stability were confirmed according to the sugar alcohol ratio. The stability analysis results are shown in Table 3 and Figures 11 to 14. In the case of items that exhibited phase separation, it was confirmed that all components did not mix but separated and coexisted in two or more liquid phases.

[0111]

[0112] Item Substance Concentration (mg / mL) Buffer Ratio of Glycerol and Mannitol pH (initial) pH (after 6 months) Properties Stability (O: Maintain properties X: Phase separation) pH Stability (O: Maintain pH, X: pH Change) Glycerol (mg / mL) Mannitol mg / mL) 1302.5mMPhosphate0306.987 37.01O73007.016.97X85mMPhosphate0306.996.95XO93276.927.04O107. 522.57.057.03O1115156.946.95O1222.57.56.966.94O132736.956.99O14 30076.95X15152.5mMPhosphate0307.016.94XO163276.947.03O177.522. 57.056.92O1815156.976.94O1922.57.57.036.93O202736.977.01O213006 .967.06X225mMPhosphate0307.046.92XO233277.066.95O247.522.576.9 4O2515157.016.97O2622.57.56.936.99O272737.017.01O283006.966.91X

[0113] * O: Maintained phase, X: Separated phase

[0114] ** O: pH maintenance, X: pH change

[0115]

[0116] In the case of the composition of the present invention, when the weight ratio of glycerol and mannitol, which are sugar alcohols, is 1 to 9: 9 to 1, it was confirmed that the liquid composition was stored for 6 months under conditions of a temperature of 25°C and a humidity of 60% without phase separation and the pH was maintained, indicating excellent stability.

[0117]

[0118] From the above description, those skilled in the art will understand that the present invention can be implemented in other specific forms without altering its technical spirit or essential characteristics. In this regard, it should be understood that the embodiments described above are illustrative in all respects and not restrictive. The scope of the present invention should be interpreted as encompassing all changes or modifications derived from the meaning and scope of the following claims and their equivalent concepts, rather than the detailed description above.

Claims

A stabilizer composition comprising 2.5 mM to 5 mM phosphate buffer, The above stabilizer composition does not contain sodium chloride, A stabilizing composition for stabilizing a liquid composition containing collagen.

2. A stabilizer composition according to claim 1, wherein the stabilizer composition further comprises a sugar alcohol.

3. A stabilizer composition according to claim 2, wherein the sugar alcohol is glycerol, mannitol or a combination thereof.

4. A stabilizer composition according to claim 3, wherein the glycerol and mannitol are in a weight ratio of 1 to 9: 9 to 1.

5. A stabilizer composition according to claim 1, wherein the stabilizer composition has a pH of 5 to 8.

6. A stabilizer composition according to claim 1, wherein the stabilizer composition prevents phase separation of the liquid composition and maintains pH. A liquid composition comprising a phosphate buffer of 2.5 mM to 5 mM and collagen, wherein the liquid composition does not contain sodium chloride.

8. A liquid composition according to claim 7, wherein collagen is contained in an amount of 1.5 to 3 wt%. A method for stabilizing a liquid composition containing collagen, comprising the step of adding a phosphate buffer of 2.5 mM to 5 mM, wherein sodium chloride is not added.

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