KR20130014150A - Liposome collecting kaempferitrin and coating carrageenan and method of the same - Google Patents
Liposome collecting kaempferitrin and coating carrageenan and method of the same Download PDFInfo
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Abstract
Description
본 발명은 캠페리트린이 포집되어 있고, 카라기난으로 코팅되어 있는 리포솜에 관한 것으로, 더욱 자세하게는 자외선 차단 효과가 있는 캠페리트린을 리포솜화하여 입자의 표면 안정성을 증가시켜 유효 물질의 피부 흡수를 최소화하고, 리포솜 외부에 천연 유래의 카라기난을 코팅하여 물질의 안정성을 높여 자외선 차단 지속시간을 증가시키는 것을 특징으로 하는 캠페리트린이 포집되어 있고, 카라기난으로 코팅되어 있는 리포솜, 이의 제조방법 및 이를 유효성분으로 함유하는 자외선 차단 화장용 조성물에 관한 것이다.
The present invention relates to liposomes in which capperitrin is trapped and coated with carrageenan, and more particularly, liposomes which have a sunscreen effect to increase the surface stability of particles to minimize skin absorption of active substances. And, by coating a carrageenan derived from the nature outside the liposomes to increase the stability of the material to increase the duration of UV protection is trapped capritririn, characterized in that the liposomes coated with carrageenan, its preparation method and active ingredient It relates to a sunscreen cosmetic composition containing.
캠페리트린(kaempferitrin)은 아프리카, 인도에서 자생하는 쌍떡잎 식물로 아욱묵 아욱과의 한해살이 풀인 양마에 다량 존재하는 플라보노이드(Flavonoid)계 물질로써 항염 효과 (Food chemistry, 117:444, 2009), 항당뇨 효과 (Bioc. Biop. Res. Comm., 380:39, 2009) 등의 연구에 대하여 알려져 있고, 한국등록특허 제10-0815476호에는 케나프(Hibiscus cannabbinus L., Malvaceae) 추출물과 이로부터 분리된 신규 캠페롤 화합물이 항산화 및 항균에 우수한 효과를 나타내는 것이 알려져 있다.Kaempferitrin is a dicotyledonous plant native to Africa and India. It is a flavonoid-based substance present in large quantities of horseshoe, a perennial herb of Mallow family, and has anti-inflammatory effects (Food chemistry, 117: 444, 2009). Effect (Bioc. Biop. Res. Comm., 380: 39, 2009), and the like, and the Korean Patent No. 10-0815476 discloses Kenaf ( Hibiscus cannabbinus L., Malvaceae) extract and isolated therefrom It is known that new camphorol compounds have excellent effects on antioxidants and antibacterial properties.
일반적으로 리포솜(Liposome)은 인지질로 구성된 전달 및 매개체로써 나노 크기(1um 이하)의 캡슐형상이며, 리포솜 내부에는 친유성과 친수성의 기능성 물질을 모두 함유시킬 수 있어 인체의 피부 세포와 유사한 생체에 적합성 물질이며 친수성 제형에 첨가하면 부유상태로 존재하게 되어 표면 안정성을 지닌다. 천연 자외선 차단제의 선행연구의 예를 살펴보면, 커피유, 대추 과일 추출물, 알로에 추출물 등이 자외선 차단효과에 효능이 있음이 알려져 있으나, 제형 내 안정성을 위한 포집 기술이나 에멀젼화 기술을 적용한 연구는 무기 자외선 차단물질을 이용한 경우를 제외하고는 전무한 실정이다.In general, liposomes are nano-sized (1 μm or less) capsules as phospholipids and mediators, and they can contain both lipophilic and hydrophilic functional materials, so they are suitable for living organisms similar to human skin cells. It is a substance that, when added to a hydrophilic formulation, remains suspended and has surface stability. Looking at the examples of previous studies of natural sunscreen, coffee oil, jujube fruit extract, aloe extract, etc. are known to be effective in the sunscreen effect, but the study applying the capture technology or emulsification technology for stability in the formulation is inorganic ultraviolet Except in the case of using a blocking material, there is no situation.
이에, 본 발명자들은 천연식물에서 추출한 캠페리트린을 리포솜화하여 안정화 및 피부표면 지속력을 높이고, 자외선 차단 효과가 있는 화장용 조성물을 제조하고자 예의 노력한 결과, 자외선 차단 효과가 있는 캠페리트린이 포집되어 있는 리포솜에 카라기난을 코팅시켜 향상된 분산 안정성을 바탕으로 자외선 차단에 효과가 있는 것을 확인하고 본 발명을 완성하게 되었다.
Therefore, the present inventors liposomes the camphorritin extracted from natural plants to increase the stabilization and skin surface sustainability, and as a result of the effort to manufacture a cosmetic composition having a sunscreen effect, as a result, the camphortrit having a sunscreen effect is collected Carrageenan is coated on the liposomes to confirm that it is effective in blocking the sun based on the improved dispersion stability to complete the present invention.
본 발명의 목적은 자외선 차단 효과가 있는 캠페리트린을 리포솜화하여 입자의 표면 안정성을 증가시켜 유효 물질의 피부 흡수를 최소화하고, 리포솜 외부에 천연 유래의 카라기난을 코팅하여 물질의 안정성을 높여 자외선 차단 지속시간을 증가시키는 캠페리트린을 포집한 리포솜, 이의 제조방법 및 이를 유효성분으로 함유하는 자외선 차단 화장용 조성물을 제공하는 데 있다.
An object of the present invention is to liposome the sunscreen to increase the surface stability of the particles by increasing the surface stability of the particles by liposomes, and to increase the stability of the material by coating the carrageenan derived from the nature outside the liposomes The present invention provides a liposome that collects camphortrin to increase its duration, a preparation method thereof, and a sunscreen cosmetic composition containing the same as an active ingredient.
상기 목적을 달성하기 위하여, 본 발명은 (a) 인지질을 혼합하여 지질용매에 용해시키는 단계; (b) 감압 진공 건조하여 인지질 막을 형성시키는 단계; (c) 캠페리트린을 버퍼에 용해시켜 제조된 수용액과 카라기난을 상기 생성된 인지질 막에 첨가한 후, 초음파 처리하여 캠페리트린을 리포솜 내로 포집하는 단계; (d) 상기 리포솜 외의 포집되지 못한 캠페리트린을 세척하여 제거하는 단계; 및 (e) 상기 세척된 리포솜 표면에 카라기난으로 필름을 형성시켜 코팅된 리포솜을 수득하는 단계를 포함하는 캠페리트린이 포집되어 있고, 카라기난으로 코팅되어 있는 리포솜의 제조방법을 제공한다.In order to achieve the above object, the present invention comprises the steps of (a) mixing the phospholipids dissolved in a lipid solvent; (b) vacuum drying to form a phospholipid membrane; (c) adding an aqueous solution and carrageenan prepared by dissolving campertrin in a buffer to the resulting phospholipid membrane, followed by sonication to capture campertrin into liposomes; (d) washing and removing non-captured camphortrins other than liposomes; And (e) forming a film with carrageenan on the washed liposome surface to obtain a coated liposome, thereby providing a method for producing a liposome that is trapped and coated with carrageenan.
본 발명은 또한, 상기의 방법으로 제조되고, 캠페리트린이 포집되어 있고, 카라기난으로 코팅되어 있는, 자외선 차단 기능을 가지는 리포솜을 제공한다.The present invention also provides a liposome having an ultraviolet blocking function, which is prepared by the above method, in which camphoritrin is collected and coated with carrageenan.
본 발명은 또한, 상기의 리포솜을 유효성분으로 함유하는 자외선 차단 화장용 조성물을 제공한다.
The present invention also provides a sunscreen cosmetic composition containing the liposome as an active ingredient.
본 발명의 리포솜화된 캠페리트린에 카라기난을 코팅함으로써 제형 내 분산 안정성을 높이고 피부침투를 최대한으로 억제하여 피부 표면 위의 유효물질 지속시간을 증가시켜 자외선 차단지수를 높인 천연 유기 자외선 차단제를 개발함으로써 캠페리트린을 제형 내 안정화 기술을 확보하고, 무기 자외선 차단제가 가지는 부작용 및 단점 극복하여 효과적인 자외선 차단 능력을 가지는 천연 유기 자외선 차단제 및 피부 외용제 조성물을 제공한다.
By coating carrageenan on the liposomeized camphortririn of the present invention, by improving the dispersion stability in the formulation and inhibiting skin penetration to the maximum, by increasing the duration of the active substance on the surface of the skin by developing a natural organic sunscreen having a high UV protection index It provides a natural organic sunscreen and external skin composition having an effective sunscreen ability by securing the stabilization technology in the formulation of camphortrin, and overcomes the side effects and disadvantages of the inorganic sunscreen.
도 1은 리포솜화된 캠페리트린의 TEM 이미지를 나타낸 것이다.1 shows a TEM image of liposomeized camphortrin.
다른 식으로 정의되지 않는 한, 본 명세서에서 사용된 모든 기술적 및 과학적 용어들은 본 발명이 속하는 기술분야에서 숙련된 전문가에 의해서 통상적으로 이해되는 것과 동일한 의미를 갖는다. 일반적으로 본 명세서에서 사용된 명명법은 본 기술분야에서 잘 알려져 있고 통상적으로 사용되는 것이다.Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In general, the nomenclature used herein is well known and commonly used in the art.
본 발명에서는 자외선 차단 효과가 있는 캠페리트린이 리포솜에 포집되어 자외선 차단물질의 피부표면에서의 제형 안정성을 극대화하여 자외선 차단 지속시간을 증가시키며, 카라기난이 리포솜 표면에 코팅막을 형성하여 UV에 의한 자외선 차단물질의 산화 안정성을 극대화시킴으로써 기존의 천연 유래의 자외선 차단제의 단점을 개선하는 자외선 차단제 화장료 조성물을 제공한다.In the present invention, the camphortririn having a sunscreen effect is trapped in the liposomes to maximize the formulation stability on the skin surface of the sunscreen material to increase the duration of the sunscreen, carrageenan forms a coating film on the liposome surface UV by UV It provides a sunscreen cosmetic composition that maximizes the oxidative stability of the blocking material to improve the disadvantages of existing natural sunscreens.
일 관점에서, 본 발명은 본 발명은 (a) 인지질을 혼합하여 지질용매에 용해시키는 단계; (b) 감압 진공 건조하여 인지질 막을 형성시키는 단계; (c) 캠페리트린을 버퍼에 용해시켜 제조된 수용액과 카라기난을 상기 생성된 인지질 막에 첨가한 후, 초음파 처리하여 캠페리트린을 리포솜 내로 포집하는 단계; (d) 상기 리포솜 외의 포집되지 못한 캠페리트린을 세척하여 제거하는 단계; 및 (e) 상기 세척된 리포솜 표면에 카라기난으로 필름을 형성시켜 코팅된 리포솜을 수득하는 단계를 포함하는 캠페리트린이 포집되어 있고, 카라기난으로 코팅되어 있는 리포솜의 제조방법에 관한 것이다.In one aspect, the present invention comprises the steps of (a) mixing the phospholipids dissolved in a lipid solvent; (b) vacuum drying to form a phospholipid membrane; (c) adding an aqueous solution and carrageenan prepared by dissolving campertrin in a buffer to the resulting phospholipid membrane, followed by sonication to capture campertrin into liposomes; (d) washing and removing non-captured camphortrins other than liposomes; And (e) forming a film with carrageenan on the washed liposome surface to obtain a coated liposome, and to a method for producing a liposome that is trapped and coated with carrageenan.
본 발명에서의 카라기난(carrageenan)은 홍조류의 Irish moss로부터 열수추출로 얻어지는 고분자의 천연 다당류로, 기본구조는 갈락토오스 중합체이고, κ-carrageenan, λ-carrageenan, ι-carrageenan 등 6형으로 대별된다. 모두 갈락토오스의 잔기로 이루어지고 있지만, 황산기의 결합상태 및 결합수가 다르다. 카라기난은 수상에서 소량으로도 용액을 점증하거나, 젤화시키는 능력을 가지고 있어 젤리화제, 약품이나 화장품의 안정화제, 분산제로서 널리 사용된다.Carrageenan (carrageenan) in the present invention is a natural polysaccharide of a polymer obtained by hot water extraction from Irish moss of red algae, the basic structure is galactose polymer, κ-carrageenan, λ-carrageenan, ι-carrageenan and roughly divided into six types. Although all consist of residues of galactose, the bond state and the number of bonds of a sulfate group differ. Carrageenan has the ability to thicken or gelate solutions even in small amounts in the water phase and is widely used as a jelly, stabilizer and dispersant for pharmaceuticals and cosmetics.
본 발명에 있어서, 상기 인지질은 포스포리피드, 포스파티딜 콜린, 리피드, 레시틴 및 콜레스테롤로 구성된 군에서 1종 이상 선택되는 것을 특징으로 할 수 있고, 상기 지질용매는 클로로포름, 에테르, 아세톤, 알코올, 클로로포름 및 이황화탄소로 구성된 군에서 선택되는 것을 특징으로 할 수 있다.In the present invention, the phospholipid may be selected from the group consisting of phospholipid, phosphatidyl choline, lipid, lecithin and cholesterol, wherein the lipid solvent is chloroform, ether, acetone, alcohol, chloroform and It may be characterized in that it is selected from the group consisting of carbon disulfide.
본 발명의 일 실시예에서는 레시틴, 콜레스테롤 혼합액을 클로로폼에 용해시킨 다음, 감압 진공 플라스크에 옮긴 후 유기용매가 증발할 때까지 진공 감압하고 40℃ 이하에서 6시간 동안 감압 농축하여 진공 플라스크 표면에 얇은 인지질 막을 형성시켰다. 그 후 PBS 버퍼에 캠페리트린이 1%가 되게 용해시킨 후 카라기난을 각각 0.5%, 1%, 2%로 첨가하여 가온 조건에서 플라스크 벽면에 형성된 인지질 막이 용해될 때까지 초음파 처리를 하여 캠페리트린을 리포솜화 하여 시료 1을 수득하였다.In an embodiment of the present invention, the lecithin and cholesterol mixture is dissolved in chloroform, and then transferred to a vacuum vacuum flask, and vacuum reduced until the organic solvent evaporates and concentrated under reduced pressure at 40 ° C. or lower for 6 hours to thin the surface of the vacuum flask. Phospholipid membranes were formed. Thereafter, dissolve 1% of camphortrin in PBS buffer, and then add carrageenan at 0.5%, 1%, and 2%, respectively, and sonicate until the phospholipid membrane formed on the flask wall is dissolved under warming conditions. Liposome was obtained to give sample 1.
본 발명에 있어서, (e)단계에서 리포솜에 카라기난 용액을 투입하여 40 ~ 60℃, 20 ~ 100hPa에서 진공 감압하고 카라기난 이외의 용매가 증발할 때까지 반복하여 리포솜 표면에 카라기난을 코팅하는 것을 특징으로 할 수 있다.In the present invention, the carrageenan solution is added to the liposome in step (e) and vacuum reduced at 40 to 60 ° C. and 20 to 100 hPa, and the carrageenan is coated on the surface of the liposome by repeating until a solvent other than carrageenan evaporates. can do.
본 발명의 다른 실시예에서는 수득한 리포솜을 PBS 버퍼를 이용하여 세척한 후 원심분리하여 15분 동안 리포솜을 가라앉힌 후, 상등액을 제거하였다. 이 세척 단계를 3회 반복하여 순수하게 리포솜화된 캠페리트린을 수득하고, 카라기난의 코팅을 위하여, 상기 수득한 리포솜에 0.5% 카라기난 수용액을 첨가하여 감압 플라스크에 넣고 60℃ 가온 조건에서 20hPa까지 진공 감압을 하여 용매가 증발하면서 리포솜 벽면에 카라기난이 코팅된 캠페리트린 함유 리포솜 시료 2를 수득하였다.In another embodiment of the present invention, the obtained liposomes were washed with PBS buffer and then centrifuged to settle the liposomes for 15 minutes, and then the supernatant was removed. This washing step was repeated three times to obtain purely liposome-formulated camphortririn, and for coating of carrageenan, 0.5% carrageenan aqueous solution was added to the obtained liposome and placed in a reduced pressure flask and vacuumed to 20 hPa at 60 ° C warming conditions. The solvent was evaporated under reduced pressure to obtain camphortririn-containing liposome sample 2 having carrageenan coated on the liposome wall.
본 발명에 있어서, (e)단계는 리포솜을 카라기난 용액으로 세척하고, 리포솜을 수득하여 냉동한 후, 이를 다시 PBS 버퍼로 세척하여 최종적으로 카라기난이 하이드로젤 상태로 코팅된 리포좀을 수득하는 것을 특징으로 할 수 있다.In the present invention, step (e) is to wash the liposomes with a carrageenan solution, obtain a liposome and freeze, and then wash it again with PBS buffer to obtain a liposome coated with carrageenan in a hydrogel state. can do.
본 발명의 또 다른 실시예에서는 수득한 리포솜을 PBS 버퍼를 이용하여 세척한 후 원심분리하여 리포솜을 가라앉힌 후 상등액을 제거하였다. 이 세척 단계를 3회 반복하여 순수하게 리포솜화된 캠페리트린을 수득하였고, 리포솜화된 캠페리트린에 카라기난 수용액을 넣은 후 voltex mixer를 이용하여 1분 동안 교반 및 원심분리하여 상등액을 제거하여 침적된 리포솜 시료 3을 수득하고, 60℃ 이상에서 건조하였다.In another embodiment of the present invention, the obtained liposomes were washed with PBS buffer and centrifuged to subside the liposomes, and then the supernatant was removed. This washing step was repeated three times to obtain purely liposome-formulated camphortrin, and the aqueous solution of carrageenan was added to the liposome-formulated camphortrin, followed by stirring and centrifugation for 1 minute using a voltex mixer to remove the supernatant. Liposome Sample 3 was obtained, and dried at 60 ° C or higher.
본 발명에 있어서, (e)단계는 리포솜에 전체적으로 카라기난 용액을 분무하고, 이를 동결건조 또는 건조하여 하이드로젤 상태로 코팅된 리포좀을 수득하는 것을 특징으로 할 수 있다.In the present invention, step (e) may be characterized by spraying the carrageenan solution to the liposome as a whole, lyophilized or dried to obtain a liposome coated in a hydrogel state.
본 발명의 또 다른 실시예에서는 수득한 리포솜을 PBS 버퍼를 이용하여 세척한 후 원심분리하여 15분 동안 리포솜을 가라앉힌 후 상등액을 제거하였다. 이 세척 단계를 3회 반복하여 순수하게 리포솜화된 캠페리트린을 수득하였다. 이후, 제조된 리포솜화된 캠페리트린에 0.5% 카라기난 수용액을 분무하여 전체적으로 카라기난 용액을 묻히게 한 후, 동결건조를 실시하여 시료 4를 수득하였다.In another embodiment of the present invention, the obtained liposomes were washed with PBS buffer and centrifuged to settle the liposomes for 15 minutes, and then the supernatant was removed. This washing step was repeated three times to obtain pure liposomes of camphoritrine. Thereafter, a 0.5% carrageenan aqueous solution was sprayed onto the prepared liposome-ized camphortririn so that the carrageenan solution was entirely buried, and then lyophilized to obtain sample 4.
다른 관점에서, 본 발명은 상기의 방법으로 제조되고, 캠페리트린이 포집되어 있고, 카라기난으로 코팅되어 있는, 자외선 차단 기능을 가지는 리포솜에 관한 것이다.In another aspect, the present invention relates to a liposome having an ultraviolet blocking function, which is prepared by the above-described method, wherein the camphortrin is collected and coated with carrageenan.
본 발명의 일 양태에서, 리포솜화 및 코팅하지 않은 캠페리트린을 함유하는 크림과 상기 제조한 시료 1~4를 함유하는 크림을 제조하여 SPF지수를 분석한 결과, 리포솜화 및 코팅하지 않은 캠페리트린을 함유하는 크림의 SPF 지수는 3.2로 나타난 반면, 리포솜화 및 코팅한 캠페리트린 함유 크림의 SPF 지수는 4.9~9.4으로 높게 나타난 것을 확인할 수 있었다.In one embodiment of the present invention, a cream containing liposomes and uncoated camphortrin and a cream containing samples 1 to 4 prepared above were analyzed and the SPF index was analyzed. The SPF index of creams containing trine was 3.2, while the SPF index of liposome-formed and creamy camphortrin-containing creams was 4.9-9.4.
또 다른 관점에서, 본 발명은 상기의 리포솜을 유효성분으로 함유하는 자외선 차단 화장용 조성물에 관한것이다.In another aspect, the present invention relates to a sunscreen cosmetic composition containing the liposome as an active ingredient.
본 발명에서 자외선 차단 효과가 있는 캠페리트린을 리포솜화하고, 카라기난을 코팅함으로써 제형 내 분산 안정성을 높이고 피부침투를 최대한으로 억제하여 피부 표면 위의 유효물질 지속시간을 증가시켜 자외선 차단지수를 높인 천연 유기 자외선 차단제를 개발할 수 있다.
In the present invention, by liposome-forming the camphortririn having a sunscreen effect, and coating the carrageenan to increase the dispersion stability in the formulation and inhibit the skin penetration to the maximum to increase the duration of the active substance on the skin surface to increase the UV block index Organic sunscreens can be developed.
이하, 실시예를 통하여 본 발명을 더욱 상세히 설명하고자 한다. 이들 실시예는 오로지 본 발명을 예시하기 위한 것으로서, 본 발명의 범위가 이들 실시예에 의해 제한되는 것으로 해석되지 않는 것은 당업계에서 통상의 지식을 가진 자에 있어서 자명할 것이다.
Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for illustrating the present invention, it will be apparent to those skilled in the art that the scope of the present invention is not to be construed as limited by these examples.
실시예 1: 캠페리트린을 함유하는 리포솜의 제조방법 Example 1 Preparation of Liposomes Containing Camparytrin
Kaempferitrin은 양마(ibiscus cannabinus L.)잎을 유기용매를 사용하여 추출 후 초고압 염석공정에 의해 유효성분인 kaempferitrin을 분리 정제하여 제조하였고, 95 % 이상의 순도를 가지는 물질을 사용하였다.Kaempferitrin was prepared by separating and purifying kaempferitrin, an active ingredient, by ultra-high pressure salting process after extracting the leaves of ibiscus cannabinus L. using an organic solvent, and using a material having a purity of 95% or more.
레시틴, 콜레스테롤 혼합액 0.5g을 클로로폼 10㎖에 용해시킨 다음, 감압 진공 플라스크에 옮긴 후 유기용매가 증발할 때까지 20hPa로 진공 감압하고 40℃ 이하에서 6시간 동안 감압 농축하여 진공 플라스크 표면에 얇은 인지질 막을 형성시켰다. 그 후 PBS 버퍼에 캠페리트린이 1%가 되게 용해시킨 후 카라기난을 각각 0.5%, 1%, 2%로 첨가하여 가온 조건(50℃)에서 플라스크 벽면에 형성된 인지질 막이 용해될 때까지 초음파 처리를 하여 캠페리트린을 리포솜화 하여 시료 1을 수득하였다.
0.5 g of lecithin and cholesterol mixture was dissolved in 10 ml of chloroform, and then transferred to a vacuum flask, and vacuum reduced to 20 hPa until the organic solvent evaporated, and concentrated under reduced pressure at 40 ° C. or lower for 6 hours to form a thin phospholipid on the surface of the vacuum flask. A film was formed. After dissolving 1% of camphortrin in PBS buffer, carrageenan was added at 0.5%, 1%, and 2%, respectively, and sonication was performed until the phospholipid membrane formed on the wall of the flask was dissolved under heating conditions (50 ° C). Was subjected to liposome formation of sample to obtain Sample 1.
실시예 2: 감압방법으로 카라기난이 코팅된 캠페리트린 함유 리포솜의 제조방법Example 2 Preparation of Carrageenan-Coated Camperritin-Containing Liposomes by Pressure Reduction Method
상기 실시예 1에서 수득한 리포솜을 PBS 버퍼를 이용하여 세척한 후 원심분리기를 이용하여 5000rpm(4,400g)에서 15분 동안 리포솜을 가라앉힌 후, 상등액을 제거하였다. 이 세척 단계를 3회 반복하여 순수하게 리포솜화된 캠페리트린을 수득하였다.The liposome obtained in Example 1 was washed with PBS buffer, and then the liposome was allowed to settle for 15 minutes at 5000 rpm (4,400 g) using a centrifuge, and then the supernatant was removed. This washing step was repeated three times to obtain pure liposomes of camphoritrine.
카라기난의 코팅을 위하여, 상기 수득한 리포솜에 0.5% 카라기난 수용액을 10㎖ 첨가하여 감압 플라스크에 넣고 60℃ 가온 조건에서 20hPa까지 진공 감압을 하여 용매가 증발하면서 리포솜 벽면에 카라기난이 코팅된 캠페리트린 함유 리포솜 시료 2를 수득하였다.
For the coating of carrageenan, 10 ml of 0.5% carrageenan aqueous solution was added to the obtained liposome and placed in a vacuum flask under reduced pressure to 20 hPa under 60 ° C. warming condition. Liposomal Sample 2 was obtained.
실시예 3: 침적방법으로 카라기난이 코팅된 캠페리트린Example 3: Carrageenan-Coated Campertrins by Immersion 함유 리포솜의 제조방법Method for producing liposomes containing
상기 실시예 1에서 수득한 리포솜을 PBS 버퍼를 이용하여 세척한 후 원심분리기를 이용하여 5000rpm(4,400g)에서 15분 동안 리포솜을 가라앉힌 후 상등액을 제거하였다. 이 세척 단계를 3회 반복하여 순수하게 리포솜화된 캠페리트린을 수득하였다. The liposomes obtained in Example 1 were washed with PBS buffer, and then the liposomes were allowed to settle for 15 minutes at 5000 rpm (4,400 g) using a centrifuge to remove the supernatant. This washing step was repeated three times to obtain pure liposomes of camphoritrine.
리포솜화된 캠페리트린에 카라기난 수용액 5㎖를 넣은 후, voltex mixer를 이용하여 1분 동안 교반 및 5000rpm(4,400g)에서 15분 동안 원심분리하여 상등액을 제거하여 침적된 리포솜 시료 3을 수득하고, 60℃ 이상에서 건조하였다.
5 ml of carrageenan solution was added to the liposome-formulated camphortrin, followed by stirring for 1 minute using a voltex mixer and centrifugation at 5000 rpm (4,400 g) for 15 minutes to remove the supernatant to obtain the deposited liposome sample 3, It dried at 60 degreeC or more.
실시예 4: 분무방법으로 카라기난이 코팅된 캠페리트린 함유 리포솜의 제조방법Example 4 Preparation of Carrageenan Coated Camperritin-Containing Liposomes by Spray Method
상기 실시예 1에서 수득한 리포솜을 PBS 버퍼를 이용하여 세척한 후 원심분리기를 이용하여 5000rpm(4,400g)에서 15분 동안 리포솜을 가라앉힌 후 상등액을 제거하였다. 이 세척 단계를 3회 반복하여 순수하게 리포솜화된 캠페리트린을 수득하였다. The liposomes obtained in Example 1 were washed with PBS buffer, and then the liposomes were allowed to settle for 15 minutes at 5000 rpm (4,400 g) using a centrifuge to remove the supernatant. This washing step was repeated three times to obtain pure liposomes of camphoritrine.
이후, 제조된 리포솜화된 캠페리트린에 0.5% 카라기난 수용액 5㎖을 분무하여 전체적으로 카라기난 용액을 묻히게 한 후, 동결건조를 실시하여 시료 4를 수득하였다.Thereafter, 5 ml of a 0.5% carrageenan aqueous solution was sprayed onto the prepared liposome-ized camphortririn, and then the whole carrageenan solution was buried, followed by lyophilization to obtain Sample 4.
제형예 1: 리포솜화 및 코팅하지 않은 캠페리트린을 함유하는 크림Formulation Example 1 Cream Containing Liposomalized and Uncoated Campertrin
리포솜화 및 코팅이 되지 않은 캠페리트린을 이용하여 크림을 제조하였다 (표 1). 구성된 성분 중에서 먼저 표 1의 원료 12 ~17의 수성성분을 80℃로 가온하여 완전 용해시킨 다음, 표 1의 원료 1~11을 80℃로 가온하여 상기 원료 12~17의 용액에 투입하고 호모믹서(Homo Mixer Mark, Primix, 일본)를 이용하여 3,000rpm에서 15분간 유화시켰다. 그 후, 원료 18을 투입하여 5분간 교반시킨 후 실온으로 냉각하였다.
Creams were prepared using liposomed and uncoated camphortrins (Table 1). Among the components, first, the aqueous components of the raw materials 12 to 17 shown in Table 1 were completely dissolved by heating to 80 ° C., and then the raw materials 1 to 11 of Table 1 were heated to 80 ° C., and added to the solution of the raw materials 12 to 17, followed by a homomixer. (Homo Mixer Mark, Primix, Japan) was used to emulsify at 3,000 rpm for 15 minutes. Then, the raw material 18 was thrown in, stirred for 5 minutes, and cooled to room temperature.
제형예 2: 리포솜화 및 코팅 처리한 캠페리트린을 함유하는 크림Formulation Example 2: Cream Containing Liposomalized and Coated Campertrins
실시예 1에서 수득한 시료 1를 이용하여, 캠페리트린 함유 리포솜을 이용하여 크림을 제조하였다 (표 2).
Using Sample 1 obtained in Example 1, a cream was prepared using camparytrin-containing liposomes (Table 2).
제형예 3: 리포솜화 및 코팅 처리한 캠페리트린을 함유하는 크림Formulation Example 3: Cream Containing Liposomalized and Coated Campertrins
실시예 2에서 수득한 시료 2를 이용하여, 카라기난이 코팅된 캠페리트린을 함유 리포솜을 이용하여 하기 제형과 같이 크림을 제조하였다 (표 3).
Using Sample 2 obtained in Example 2, a cream was prepared using the liposome containing carrageenan coated camphortrin as shown in the following formulation (Table 3).
제형예 4: 리포솜화 및 코팅 처리한 캠페리트린을 함유하는 크림Formulation Example 4: Cream Containing Liposome and Coated Campertrin
실시예 3에서 수득한 시료 3을 이용하여, 카라기난이 코팅된 캠페리트린을 함유 리포솜을 이용하여 하기 제형과 같이 크림을 제조하였다 (표 4).
Using Sample 3 obtained in Example 3, a cream was prepared using liposomes containing carrageenan coated camphortrin as shown in the following formulation (Table 4).
제형예 5: 리포솜화 및 코팅 처리한 캠페리트린을 함유하는 크림Formulation Example 5: Cream Containing Liposomalized and Coated Campertrins
실시예 4에서 수득한 시료 4를 이용하여, 카라기난이 코팅된 캠페리트린을 함유 리포솜을 이용하여 하기 제형과 같이 크림을 제조하였다 (표 5).
Using Sample 4 obtained in Example 4, a cream was prepared using the liposome containing carrageenan coated camphortrin as shown in the following formulation (Table 5).
시험예 1: 리포솜화된 캠페리트린의 TEM 촬영 및 Test Example 1: TEM imaging of liposome-ized camphortririn and in vitroin vitro 에서의 SPF 측정 SPF measurement at
상기 실시예 1에서 제조된 시료 1의 TEM 촬영하였고 (도 1), Optometrics turnkey system 290S in vitro SPF 측정 장비를 사용하여 상기 실시예 1~4에서 제조한 시료 1~4의 자외선 차단능을 시험하였고, 대조군으로 리포솜화 및 코팅되지않은 캠페리트린을 사용하여 제조한 크림을 제형예 1에 나타내었고, 리포솜화된 캠페리트린을 사용하여 제형예 2~5로 나타내었다. 그 결과, 리포솜화 및 코팅하여 제조한 캠페리트린을 함유하는 크림의 SPF 지수가 리포솜화 및 코팅하지 않은 캠페리트린 함유 크림보다 높게 나타나는 것을 확인할 수 있었다 (표 6).
TEM of Sample 1 prepared in Example 1 was taken (FIG. 1), and the UV blocking ability of Samples 1 to 4 prepared in Examples 1 to 4 was tested using an Optometrics turnkey system 290S in vitro SPF measuring apparatus. , A cream prepared using liposomes and uncoated camphortrin as a control group is shown in Formulation Example 1, and a liposomeized camphortrinin is shown as Formulation Examples 2 to 5. As a result, it was confirmed that the SPF index of the cream containing camphortririn prepared by liposomeization and coating was higher than that of the camphorritrin-containing cream without liposomeization and coating (Table 6).
이상으로 본 발명 내용의 특정한 부분을 상세히 기술하였는바, 당업계의 통상의 지식을 가진 자에게 있어서, 이러한 구체적 기술은 단지 바람직한 실시양태일 뿐이며, 이에 의해 본 발명의 범위가 제한되는 것이 아닌 점은 명백할 것이다. 따라서 본 발명의 실질적인 범위는 첨부된 청구항들과 그것들의 등가물에 의하여 정의된다고 할 것이다.The specific parts of the present invention have been described in detail above, and it is apparent to those skilled in the art that such specific descriptions are merely preferred embodiments, and thus the scope of the present invention is not limited thereto. something to do. It is therefore intended that the scope of the invention be defined by the claims appended hereto and their equivalents.
Claims (8)
(a) 인지질을 혼합하여 지질용매에 용해시키는 단계;
(b) 감압 진공 건조하여 인지질 막을 형성시키는 단계;
(c) 캠페리트린을 버퍼에 용해시켜 제조된 수용액과 카라기난을 상기 생성된 인지질 막에 첨가한 후, 초음파 처리하여 캠페리트린을 리포솜 내로 포집하는 단계;
(d) 상기 리포솜에 포집되지 못한 캠페리트린을 세척하여 제거하는 단계; 및
(e) 상기 세척된 리포솜 표면에 카라기난으로 필름을 형성시켜 코팅된 리포솜을 수득하는 단계.
A method for preparing a liposome coated with carrageenan, wherein the camphoritrine is collected, comprising the following steps:
(a) mixing the phospholipids and dissolving them in a lipid solvent;
(b) vacuum drying to form a phospholipid membrane;
(c) adding an aqueous solution and carrageenan prepared by dissolving campertrin in a buffer to the resulting phospholipid membrane, followed by sonication to capture campertrin into liposomes;
(d) washing and removing the camphortrin that was not collected in the liposomes; And
(e) forming a film with carrageenan on the washed liposome surface to obtain a coated liposome.
The method of claim 1, wherein the phospholipid is at least one selected from the group consisting of phospholipid, phosphatidyl choline, lipid, lecithin and cholesterol.
The method of claim 1, wherein the lipid solvent is selected from the group consisting of chloroform, ether, acetone, alcohol, chloroform and carbon disulfide.
According to claim 1, step (e) is a carrageenan solution is added to the liposome, the vacuum pressure at 40 ~ 60 ℃, 20 ~ 100hPa and repeated until the solvent other than carrageenan evaporates to coat the carrageenan on the liposome surface How to.
The method according to claim 1, wherein step (e) washes the liposomes with a carrageenan solution, obtains and freezes the liposomes, and washes them again with PBS buffer to finally obtain liposomes coated with carrageenan in a hydrogel state. How to.
The method of claim 1, wherein step (e) comprises spraying the carrageenan solution on the liposome as a whole, and lyophilizing or drying it to obtain a liposome coated in a hydrogel state.
A liposome having an ultraviolet blocking function, which is prepared by the method of any one of claims 1 to 7, wherein camphoritrin is collected and coated with carrageenan.
A sunscreen cosmetic composition comprising the liposome of claim 7 as an active ingredient.
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CN109007779A (en) * | 2018-06-13 | 2018-12-18 | 福建农林大学 | A kind of frozen minced fillets compounding water-retaining agent and its preparation method and application |
CN109007776A (en) * | 2018-06-13 | 2018-12-18 | 福建农林大学 | A kind of Chinese style meat dish compounding water-retaining agent and its preparation method and application |
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CN109007779A (en) * | 2018-06-13 | 2018-12-18 | 福建农林大学 | A kind of frozen minced fillets compounding water-retaining agent and its preparation method and application |
CN109007776A (en) * | 2018-06-13 | 2018-12-18 | 福建农林大学 | A kind of Chinese style meat dish compounding water-retaining agent and its preparation method and application |
CN109007779B (en) * | 2018-06-13 | 2021-07-23 | 福建农林大学 | Frozen minced fillet compound water-retaining agent and preparation method and application thereof |
CN109007776B (en) * | 2018-06-13 | 2021-08-03 | 福建农林大学 | Compound water-retaining agent for Chinese meat dishes as well as preparation method and application of compound water-retaining agent |
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