KR20220149091A - A pharmaceutical composition for preventing or treating osteoporosis comprising MSM, calcium and rosemary extract as an active ingredient - Google Patents
A pharmaceutical composition for preventing or treating osteoporosis comprising MSM, calcium and rosemary extract as an active ingredient Download PDFInfo
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- KR20220149091A KR20220149091A KR1020210056276A KR20210056276A KR20220149091A KR 20220149091 A KR20220149091 A KR 20220149091A KR 1020210056276 A KR1020210056276 A KR 1020210056276A KR 20210056276 A KR20210056276 A KR 20210056276A KR 20220149091 A KR20220149091 A KR 20220149091A
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- South Korea
- Prior art keywords
- extract
- calcium
- rosemary
- preventing
- pharmaceutical composition
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Abstract
Description
본 발명은 메틸설포닐메탄, 칼슘 및 로즈마리 추출물을 유효성분으로 포함하는 골다공증 예방 또는 치료용 약학 조성물에 관한 것이다. The present invention relates to a pharmaceutical composition for preventing or treating osteoporosis comprising methylsulfonylmethane, calcium and rosemary extract as active ingredients.
식물 유래의 다양한 화학물질들은 항 골다공증 작용, 파골세포 분화 억제 작용과 세포보호 작용을 포함한 여러 가지 생리활성들을 나타내어 건강기능식품의 유용한 소재로서 각광을 받고 있다. Various chemical substances derived from plants exhibit various physiological activities, including anti-osteoporosis, osteoclast differentiation, and cytoprotection, and are in the spotlight as useful materials for health functional foods.
한편 국내뿐 아니라 전 세계적으로 고령화, 운동 부족, 흡연 등에 의해 증가하고 있는 골다공증은 다발성 골절, 고관절 골절 등 심각한 위험을 초래하는 질병으로서, 노년기의 삶의 질을 크게 저하시키기 때문에 이를 예방하고 치료하기 위한 생리활성물질의 개발이 요구된다. On the other hand, osteoporosis, which is increasing not only in Korea but also worldwide due to aging, lack of exercise, and smoking, is a disease that causes serious risks such as multiple fractures and hip fractures. The development of bioactive substances is required.
이와 관련하여 한국등록특허 제10-1279852호는 하나 이상의 칼슘 보충제 및 하나 이상의 마그네슘 보충제를 포함하는 무기질 혼합물; 및 스트렙토코쿠스 서모필루스 ST3(Streptococcus thermophilus ST3)(KCTC 11870BP); 락토바실루스 람노수스 LR3 (Lactobacillus rhamnosus LR3)(KCTC 11868BP); 및 락토바실루스 파라카제이 LPC4(Lactobacillus paracasei LPC4) (KCTC 11866BP)를 포함하는 골다공증 예방 또는 치료용 조성물을 개시하고 있다. In this regard, Korean Patent Registration No. 10-1279852 discloses an inorganic mixture comprising one or more calcium supplements and one or more magnesium supplements; and Streptococcus thermophilus ST3 (KCTC 11870BP); Lactobacillus rhamnosus LR3 (KCTC 11868BP); And Lactobacillus paracasei LPC4 (Lactobacillus paracasei LPC4) (KCTC 11866BP) discloses a composition for preventing or treating osteoporosis, including.
또한 한국공개특허 제10-2004-0088976호는 (a) 소뼈, 오징어뼈, 홍조류, 유기산, 및 정제수를 포함하여 구성되는 미네랄 보급용 알칼리 수용액; (b) 생약 조성물; (c) 황태; (d) 검정콩; (e) 표고버섯; (f) casein phospeptide; 및 (g) DHA(Docosahexaenoic acid)를 포함하여 구성되는 골다공증 예방 및 개선용 조성물을 개시하고 있다. In addition, Korean Patent Laid-Open Publication No. 10-2004-0088976 discloses (a) an aqueous alkali solution for mineral replenishment comprising bovine bones, squid bones, red algae, organic acids, and purified water; (b) herbal compositions; (c) Hwangtae; (d) black beans; (e) shiitake mushrooms; (f) casein phospeptide; And (g) it discloses a composition for preventing and improving osteoporosis comprising DHA (Docosahexaenoic acid).
아울러 한국공개특허 제10-2012-0001283호는 갈근, 마황, 건강, 대조, 계피, 작약 및 감초를 포함하는 생약 혼합물에 물 또는 유기용매를 가한 후 추출하여 생약 추출물을 제조하고, 상기 생약 추출물에 유산균을 접종하여 발효시킴으로써 제조되는 것을 특징으로 하는 생약 추출물의 유산균 발효물을 유효성분으로 함유하는 골다공증 예방 또는 치료용 약학적 조성물을 개시하고 있다. In addition, Korea Patent Laid-Open Patent No. 10-2012-0001283 discloses that a herbal extract is prepared by adding water or an organic solvent to a herbal mixture containing ginseng root, ephedra, health, coriander, cinnamon, peony and licorice, and then extracting the extract. Disclosed is a pharmaceutical composition for preventing or treating osteoporosis containing a fermented product of lactic acid bacteria of a herbal extract, characterized in that it is prepared by inoculating and fermenting lactic acid bacteria as an active ingredient.
그러나 상기 문헌에 개시된 기술은 무기질 혼합물, 알칼리 수용액 또는 생약 추출물을 활용한 것으로서, 제조방법이 복잡하고 약효가 우수하지 않다. However, the technique disclosed in the above document utilizes an inorganic mixture, an aqueous alkali solution or a herbal extract, and the manufacturing method is complicated and the medicinal effect is not excellent.
또한 장기간 복용 시 체중 증가, 두통, 우울증 등의 부작용을 초래하는 문제점이 있다. In addition, there is a problem that causes side effects such as weight gain, headache, and depression when taken for a long time.
본 발명은 상기 종래 기술의 문제점을 해결하기 위한 것으로서, 쉽고 간단하게 제조할 수 있으며 약효가 우수한 골다공증 예방 또는 치료용 약학 조성물을 제공하는 것을 목적으로 한다. An object of the present invention is to provide a pharmaceutical composition for preventing or treating osteoporosis that can be easily and simply prepared and has excellent medicinal efficacy.
또한 본 발명은 장기간 복용 시 체중 증가, 두통 등의 부작용을 최소화할 수 있는 골다공증 예방 또는 치료용 약학 조성물을 제공하는데 그 목적이 있다. Another object of the present invention is to provide a pharmaceutical composition for preventing or treating osteoporosis that can minimize side effects such as weight gain and headache when taken for a long time.
상기와 같은 목적을 달성하기 위하여 본 발명은 메틸설포닐메탄, 칼슘 및 로즈마리 추출물을 유효성분으로 포함하는 골다공증 예방 또는 치료용 약학 조성물을 제공한다. In order to achieve the above object, the present invention provides a pharmaceutical composition for preventing or treating osteoporosis comprising methylsulfonylmethane, calcium and rosemary extract as active ingredients.
본 발명의 일실시예에 있어서, 상기 로즈마리 추출물은 로즈마리를 용매로 가열 추출하고 감압 농축하여 수득하는 것을 특징으로 한다. In one embodiment of the present invention, the rosemary extract is characterized in that it is obtained by heating and extracting rosemary with a solvent and concentration under reduced pressure.
본 발명의 일실시예에 있어서, 상기 로즈마리 추출물은 로즈마리를 용매로 가열 추출하고 감압 농축하여 용매를 제거한 후, 정제수로 수용화한 수용성 추출물인 것을 특징으로 한다. In one embodiment of the present invention, the rosemary extract is a water-soluble extract obtained by heat-extracting rosemary with a solvent, removing the solvent by concentration under reduced pressure, and then soluble in purified water.
본 발명의 일실시예에 있어서, 상기 조성물은 메틸설포닐메탄 100중량부에 대하여 칼슘 5~30중량부 및 로즈마리 추출물 2~15중량부를 포함하는 것을 특징으로 한다. In one embodiment of the present invention, the composition is characterized in that it contains 5 to 30 parts by weight of calcium and 2 to 15 parts by weight of rosemary extract based on 100 parts by weight of methylsulfonylmethane.
또한 본 발명은 메틸설포닐메탄, 칼슘 및 로즈마리 추출물을 유효성분으로 포함하는 골다공증 예방 또는 개선용 식품 조성물을 제공한다. In addition, the present invention provides a food composition for preventing or improving osteoporosis comprising methylsulfonylmethane, calcium and rosemary extract as active ingredients.
본 발명은 쉽고 간단하게 제조할 수 있으며 약효가 우수한 골다공증 예방 또는 치료용 약학 조성물을 제공할 수 있다. The present invention can provide a pharmaceutical composition for preventing or treating osteoporosis that can be easily and simply prepared and has excellent medicinal efficacy.
또한 본 발명은 장기간 복용 시 체중 증가, 두통 등의 부작용을 최소화할 수 있는 골다공증 예방 또는 치료용 약학 조성물을 제공할 수 있다. In addition, the present invention can provide a pharmaceutical composition for preventing or treating osteoporosis that can minimize side effects such as weight gain and headache when taken for a long period of time.
도 1은 본 발명의 조성물을 섭취한 경우 실험동물의 체중변화를 나타낸다.
도 2는 본 발명의 조성물을 섭취한 경우 실험동물의 대퇴부 뼈의 골밀도를 나타낸다.
도 3은 본 발명의 조성물을 섭취한 경우 실험동물의 대퇴부 뼈의 무기질 함량을 나타낸다. 1 shows the change in body weight of experimental animals when the composition of the present invention is ingested.
Figure 2 shows the bone density of the femoral bone of the experimental animal when the composition of the present invention is ingested.
Figure 3 shows the mineral content of the femoral bone of the experimental animal when the composition of the present invention is ingested.
이하 실시예를 바탕으로 본 발명을 상세히 설명한다. 본 발명에 사용된 용어, 실시예 등은 본 발명을 보다 구체적으로 설명하고 통상의 기술자의 이해를 돕기 위하여 예시된 것에 불과할 뿐이며, 본 발명의 권리범위 등이 이에 한정되어 해석되어서는 안 된다. The present invention will be described in detail based on the following examples. The terms, examples, etc. used in the present invention are merely exemplified to explain the present invention in more detail and help those of ordinary skill in the art to understand, and the scope of the present invention should not be construed as being limited thereto.
본 발명에 사용되는 기술 용어 및 과학 용어는 다른 정의가 없다면 이 발명이 속하는 기술 분야에서 통상의 지식을 가진 자가 통상적으로 이해하고 있는 의미를 나타낸다. Technical terms and scientific terms used in the present invention represent meanings commonly understood by those of ordinary skill in the art to which this invention belongs, unless otherwise defined.
본 발명은 메틸설포닐메탄, 칼슘 및 로즈마리 추출물을 유효성분으로 포함하는 골다공증 예방 또는 치료용 약학 조성물에 관한 것이다. The present invention relates to a pharmaceutical composition for preventing or treating osteoporosis comprising methylsulfonylmethane, calcium and rosemary extract as active ingredients.
상기 메틸설포닐메탄(Methyl sulfonyl methane; MSM)은 조골세포의 분화를 촉진하여 관절 및 뼈 건강에 도움을 줄 수 있다. The methyl sulfonyl methane (Methyl sulfonyl methane; MSM) can help joint and bone health by promoting the differentiation of osteoblasts.
상기 칼슘은 골다공증 및 관절질환을 예방하고 개선하는 성분으로서, 해조칼슘, 코랄칼슘, 어골칼슘, 탄산칼슘, 인산칼슘, 난각칼슘, 진주칼슘, 패각칼슘 등이 사용될 수 있으며, 분말, 액상, 이온, 염, 조성물, 복합체 등 다양한 형태로 사용될 수 있다. The calcium is a component for preventing and improving osteoporosis and joint diseases, and seaweed calcium, coral calcium, fish bone calcium, calcium carbonate, calcium phosphate, egg shell calcium, pearl calcium, shell calcium, etc. may be used, and powder, liquid, ion, It may be used in various forms, such as salts, compositions, and complexes.
상기 로즈마리 추출물은 높은 peroxyl radical 소거 활성 및 reducing capacity를 나타내고, 세포 내 산화 스트레스를 효과적으로 감소시켜 우수한 세포 내 항산화 활성을 나타내며, 높은 파골세포 분화 억제 활성을 가지므로 골다공증의 예방에 효과가 있다. The rosemary extract exhibits high peroxyl radical scavenging activity and reducing capacity, exhibits excellent intracellular antioxidant activity by effectively reducing intracellular oxidative stress, and has high osteoclast differentiation inhibitory activity, so it is effective in preventing osteoporosis.
상기 로즈마리 추출물은 로즈마리를 물, 알코올(에탄올, 메탄올, 주정, 발효주정 등), 아세톤 등의 용매로 가열 추출하고 감압 농축하여 수득할 수 있다. The rosemary extract can be obtained by heating and extracting rosemary with a solvent such as water, alcohol (ethanol, methanol, alcohol, fermented alcohol, etc.), acetone, and then concentrating under reduced pressure.
예를 들면, 상기 로즈마리 추출물은 로즈마리를 알코올로 가열 추출하고 감압 농축하여 수득한 알코올 추출물인 것이 바람직하다. For example, the rosemary extract is preferably an alcohol extract obtained by heating and extracting rosemary with alcohol and concentrating under reduced pressure.
또한 상기 로즈마리 추출물은 로즈마리를 알코올로 가열 추출하고 감압 농축하여 알코올을 제거한 후, 정제수로 수용화한 수용성 추출물을 사용할 수 있다. In addition, the rosemary extract may be used as a water-soluble extract obtained by heating and extracting rosemary with alcohol and concentrating under reduced pressure to remove the alcohol, and then soluble in purified water.
상기 로즈마리 수용성 추출물은 물에 잘 녹고 점도가 낮아 섭취가 간편하고, 상기 수용화 과정을 거치면서 일부 유효성분의 함량이 증가할 수 있다. The water-soluble extract of rosemary is easily soluble in water and has a low viscosity, so it is easy to ingest, and the content of some active ingredients may increase through the water-soluble process.
가열 추출은 로즈마리 100중량부에 대하여 알코올 100~1,000중량부를 가하고 40~90℃에서 1~12시간 가열하여 추출할 수 있으며, 알코올로 1~3시간씩 1~5번 반복해서 가열 추출하는 것이 바람직하다. Heat extraction can be performed by adding 100 to 1,000 parts by weight of alcohol to 100 parts by weight of rosemary and heating at 40 to 90° C. for 1 to 12 hours. It is preferable to heat extraction with alcohol repeatedly 1 to 5 times for 1 to 3 hours each. do.
가열 추출한 후 감압 농축하여 로즈마리 알코올 추출물을 수득할수 있다. After extraction by heating, it can be concentrated under reduced pressure to obtain a rosemary alcohol extract.
또한 로즈마리를 알코올로 가열 추출하고 감압 농축하여 알코올을 제거한 후, 이를 다시 정제수로 수용화함으로써, 로즈마리 수용성 추출물을 제조할 수 있다. In addition, rosemary water-soluble extract can be prepared by heat-extracting rosemary with alcohol and concentrating under reduced pressure to remove the alcohol, and then re-solubilizing it with purified water.
상기 로즈마리 수용성 추출물은 그대로 사용하거나 감압 농축하여 정제수를 제거하여 사용할 수도 있다. The water-soluble extract of rosemary may be used as it is or concentrated under reduced pressure to remove purified water.
본 발명은 상기 로즈마리 추출물로서 알코올 추출물과 수용성 추출물을 동시에 사용할 수도 있고, 이때 상기 알코올 추출물 및 수용성 추출물의 중량비는 60~80:20~40 인 것이 바람직하다. 상기 알코올 추출물 및 수용성 추출물의 중량비가 상기 수치범위를 만족하는 경우, 골다공증 예방효과가 극대화될 수 있다. In the present invention, an alcohol extract and a water-soluble extract may be used simultaneously as the rosemary extract, and in this case, the weight ratio of the alcohol extract and the water-soluble extract is preferably 60 to 80: 20 to 40. When the weight ratio of the alcohol extract and the water-soluble extract satisfies the numerical range, the osteoporosis prevention effect can be maximized.
본 발명의 조성물은 메틸설포닐메탄 100중량부에 대하여 칼슘 5~30중량부 및 로즈마리 추출물 2~15중량부를 포함할 수 있다. The composition of the present invention may include 5 to 30 parts by weight of calcium and 2 to 15 parts by weight of rosemary extract based on 100 parts by weight of methylsulfonylmethane.
상기 메틸설포닐메탄, 칼슘 및 로즈마리 추출물의 함량이 상기 수치범위를 만족하는 경우, 골다공증 예방효과가 극대화될 수 있다. When the content of the methylsulfonylmethane, calcium and rosemary extract satisfies the above numerical range, the osteoporosis prevention effect can be maximized.
또한 본 발명의 조성물은 프로폴리스 추출물을 추가로 포함할 수 있다. In addition, the composition of the present invention may further include a propolis extract.
상기 프로폴리스 추출물은 높은 peroxyl radical 소거 활성 및 reducing capacity를 나타내고, 세포 내 산화 스트레스를 효과적으로 감소시켜 우수한 세포 내 항산화 활성을 나타내며, 높은 파골세포 분화 억제 활성을 가지므로 골다공증의 예방에 효과가 있다. The propolis extract exhibits high peroxyl radical scavenging activity and reducing capacity, exhibits excellent intracellular antioxidant activity by effectively reducing intracellular oxidative stress, and has high osteoclast differentiation inhibitory activity, so it is effective in preventing osteoporosis.
상기 프로폴리스 추출물은 프로폴리스를 에탄올, 메탄올, 주정, 발효주정 등의 알코올로 가열 추출하고 감압 농축하여 수득한다. The propolis extract is obtained by heating and extracting propolis with alcohol, such as ethanol, methanol, alcohol, or fermented alcohol, and concentrating under reduced pressure.
예를 들면, 상기 프로폴리스 추출물은 프로폴리스를 에탄올로 가열 추출하고 감압 농축하여 수득한 알코올 추출물인 것이 바람직하다. For example, the propolis extract is preferably an alcohol extract obtained by heat-extracting propolis with ethanol and concentration under reduced pressure.
또한 상기 프로폴리스 추출물은 프로폴리스를 알코올로 가열 추출하고 감압 농축하여 알코올을 제거한 후, 정제수로 수용화한 수용성 추출물을 사용할 수 있다. In addition, the propolis extract may be an aqueous extract obtained by heating and extracting propolis with alcohol and concentrating under reduced pressure to remove the alcohol, and then soluble in purified water.
가열 추출은 프로폴리스 100중량부에 대하여 알코올 100~1,000중량부를 가하고 40~90℃에서 1~12시간 가열하여 추출할 수 있으며, 알코올로 1~3시간씩 1~5번 반복해서 가열 추출하는 것이 바람직하다. Heat extraction can be performed by adding 100 to 1,000 parts by weight of alcohol to 100 parts by weight of propolis and heating at 40 to 90°C for 1 to 12 hours. desirable.
가열 추출한 후 감압 농축하여 프로폴리스 알코올 추출물을 수득한다. After heat extraction, the mixture was concentrated under reduced pressure to obtain a propolis alcohol extract.
또한 프로폴리스를 알코올로 가열 추출하고 감압 농축하여 알코올을 제거한 후, 이를 다시 정제수로 수용화함으로써, 프로폴리스 수용성 추출물을 제조할 수 있다. In addition, by heating and extracting propolis with alcohol and concentrating under reduced pressure to remove the alcohol, and re-solubilizing it with purified water, a water-soluble propolis extract can be prepared.
상기 프로폴리스 수용성 추출물은 그대로 사용하거나 감압 농축하여 정제수를 제거하여 사용할 수도 있다. The aqueous propolis extract may be used as it is or concentrated under reduced pressure to remove purified water.
본 발명은 상기 프로폴리스 추출물로서 알코올 추출물과 수용성 추출물을 동시에 사용할 수도 있고, 이때 상기 알코올 추출물 및 수용성 추출물의 중량비는 20~40:60~80 인 것이 바람직하다. 상기 알코올 추출물 및 수용성 추출물의 중량비가 상기 수치범위를 만족하는 경우, 골다공증 예방효과가 극대화될 수 있다. In the present invention, an alcohol extract and a water-soluble extract may be used simultaneously as the propolis extract, and in this case, the weight ratio of the alcohol extract and the water-soluble extract is preferably 20-40:60-80. When the weight ratio of the alcohol extract and the water-soluble extract satisfies the numerical range, the osteoporosis prevention effect can be maximized.
상기 프로폴리스 추출물은 메틸설포닐메탄 100중량부에 대하여 2~10중량부 사용되는 것이 바람직하며, 프로폴리스 추출물의 함량이 상기 수치범위를 만족하는 경우, 골다공증 예방효과가 극대화될 수 있다. The propolis extract is preferably used in an amount of 2 to 10 parts by weight based on 100 parts by weight of methylsulfonylmethane.
또한 로즈마리 추출물 및 프로폴리스 추출물의 중량비는 60~80:20~40 인 것이 바람직하다. 상기 로즈마리 추출물 및 프로폴리스 추출물의 중량비가 상기 수치범위를 만족하는 경우, 골다공증 예방효과가 극대화될 수 있다. In addition, the weight ratio of the rosemary extract and the propolis extract is preferably 60 to 80: 20 to 40. When the weight ratio of the rosemary extract and the propolis extract satisfies the numerical range, the osteoporosis prevention effect can be maximized.
본 발명의 약학 조성물은 약학적 조성물의 제조에 통상적으로 사용하는 적절한 담체, 부형제 및 희석제를 더 포함할 수 있다. The pharmaceutical composition of the present invention may further include suitable carriers, excipients and diluents commonly used in the preparation of pharmaceutical compositions.
상기 담체, 부형제 및 희석제로는 락토즈, 텍스트로즈, 수크로스, 솔비톨, 만니톨, 자일리톨, 에리스리톨, 말티톨, 전분, 아카시아 고무, 알지네이트, 젤라틴, 칼슘 포스페이트, 칼슘 실리케이트, 셀롤로즈, 메틸 셀롤로즈, 미정질 셀롤로즈, 폴리비닐 피톨리돈, 물, 메틸히드록시벤조에이트, 프로필 히드록시벤조에이트, 탈크, 마그네슘 스테아레이트 및 광물유를 들 수 있다. The carrier, excipient and diluent include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, unknown. vaginal cellulose, polyvinyl phytolidone, water, methylhydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate and mineral oil.
본 발명의 약학 조성물은 산제, 과립제, 정제, 캡슐제, 현탁액, 에멀젼, 시럽, 에어로졸 등의 경구형 제형, 외용제, 좌제 및 멸균 주사용액의 형태로 제형화하여 사용될 수 있다. The pharmaceutical composition of the present invention may be formulated and used in the form of oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories, and sterile injection solutions.
또한 본 발명은 메틸설포닐메탄, 칼슘 및 로즈마리 추출물을 유효성분으로 포함하는 골다공증 예방 또는 개선용 식품 조성물에 관한 것이다. The present invention also relates to a food composition for preventing or improving osteoporosis comprising methylsulfonylmethane, calcium and rosemary extract as active ingredients.
식품은 각종 식품류, 캔디, 음료, 껌, 차, 비타민 복합제, 건강 기능성 식품류 등의 형태일 수 있고, 분말, 과립, 정제, 캡슐, 음료 등의 형태로 제공될 수 있다. Food may be in the form of various foods, candy, beverages, gum, tea, vitamin complexes, health functional foods, and the like, and may be provided in the form of powders, granules, tablets, capsules, beverages, and the like.
이하 실시예를 통해 본 발명을 상세히 설명한다. 하기 실시예는 본 발명의 실시를 위하여 예시된 것일 뿐, 본 발명의 내용이 하기 실시예에 의하여 한정되는 것은 아니다. The present invention will be described in detail with reference to the following examples. The following examples are only exemplified for the practice of the present invention, and the content of the present invention is not limited by the following examples.
(실시예 1) 로즈마리 추출물의 제조 (Example 1) Preparation of rosemary extract
건조된 로즈마리 100g 을 95% 에탄올로 50℃에서 3회 환류 추출한 다음 감압 농축하여 추출물을 수득하였다(로즈마리 알코올 추출물). 100 g of dried rosemary was extracted three times under reflux with 95% ethanol at 50° C., and then concentrated under reduced pressure to obtain an extract (rosemary alcohol extract).
건조된 로즈마리 100g 을 95% 에탄올로 50℃에서 3회 환류 추출한 다음 감압 농축하고 용매를 모두 제거한 후, 정제수로 수용화시켜 추출물을 수득하였다(로즈마리 수용성 추출물). 100 g of dried rosemary was extracted three times under reflux at 50° C. with 95% ethanol, concentrated under reduced pressure, and all solvents were removed, and then aqueous solution was obtained with purified water to obtain an extract (water-soluble rosemary extract).
또한 상기 로즈마리 추출물의 제조방법과 동일한 방법으로 프로폴리스 추출물을 제조하였다. In addition, a propolis extract was prepared in the same manner as in the preparation method of the rosemary extract.
(실시예 2) 조성물의 TRAP 활성 (Example 2) TRAP activity of the composition
Tartrate Resistant Acid Phosphatase(TRAP) 활성은 RAW 264.7 세포를 96-well plate에 1×104 cells/well의 수로 24시간 예비 배양하고, 분화유도배지 [50 ng/mL receptor activator of nuclear factor-κB ligand (RANKL)]와 함께 시료를 농도별로 처리하였다. Tartrate Resistant Acid Phosphatase (TRAP) activity was pre-incubated with RAW 264.7 cells in a 96-well plate at the number of 1×10 4 cells/well for 24 hours, and differentiation induction medium [50 ng/mL receptor activator of nuclear factor-κB ligand ( RANKL)] and the samples were treated by concentration.
매 2일마다 배지 교환 시에 시료를 넣고 총 5일간 배양하였다. Samples were added at the time of medium exchange every 2 days and cultured for a total of 5 days.
Media를 제거한 후 50 μL의 PBS (pH 7.4)로 1회 세척 후 10 mM sodium tartrate와 5mM ρ-nitrophenylphosphate가 포함된 50 mM citrate buffer를 100 μL씩 분주한 후 37℃에서 1시간 반응하였다. After removing the media, after washing once with 50 μL of PBS (pH 7.4), 100 μL of 50 mM citrate buffer containing 10 mM sodium tartrate and 5 mM ρ-nitrophenylphosphate was dispensed at a time and reacted at 37° C. for 1 hour.
반응 후 0.1 N NaOH로 반응을 중지시켜 405 nm에서 흡광도를 측정하였다. After the reaction, the reaction was stopped with 0.1 N NaOH and absorbance was measured at 405 nm.
시료의 RAW 264.7 세포 주에서 파골세포 분화 유도인자인 RANKL에 의해 5일간 분화시킨 후 파골세포 특이적으로 발현되는 Tartrate Resistant Acid Phosphatase (TRAP)의 활성을 측정한 결과는 표 1에 제시된다. Table 1 shows the results of measuring the activity of Tartrate Resistant Acid Phosphatase (TRAP) specifically expressed in osteoclasts after 5 days of differentiation by RANKL, an osteoclast differentiation inducer, in the RAW 264.7 cell line of the sample.
(100:20)Methylsulfonylmethane + Calcium
(100:20)
(100:20:10)Methylsulfonylmethane + Calcium + Rosemary Extract
(100:20:10)
(100:20:1)Methylsulfonylmethane + Calcium + Rosemary Extract
(100:20:1)
(100:20:20)Methylsulfonylmethane + Calcium + Rosemary Extract
(100:20:20)
(100:20:10:5)Methylsulfonylmethane + Calcium + Rosemary Extract + Propolis Extract
(100:20:10:5)
(100:20:5:5)Methylsulfonylmethane + Calcium + Rosemary Extract + Propolis Extract
(100:20:5:5)
(100:20:14:3)Methylsulfonylmethane + Calcium + Rosemary Extract + Propolis Extract
(100:20:14:3)
메틸설포닐메탄, 칼슘 및 로즈마리 추출물을 사용하는 경우 또는 메틸설포닐메탄, 칼슘, 로즈마리 추출물 및 프로폴리스 추출물을 사용하는 경우, RANKL에 의해 유도된 TRAP의 활성을 효과적으로 억제시키는 것으로 나타났다. When methylsulfonylmethane, calcium and rosemary extracts were used, or when methylsulfonylmethane, calcium, rosemary extracts and propolis extracts were used, it was shown that the activity of TRAP induced by RANKL was effectively inhibited.
이러한 결과로부터, 로즈마리 추출물 및/또는 프로폴리스 추출물은 파골세포의 형성과 TRAP 활성을 효과적으로 억제시켜 뼈의 흡수를 방해함으로써 뼈 건강에 긍정적 효과를 나타낼 수 있을 것으로 판단된다. From these results, it is determined that the rosemary extract and/or propolis extract can have a positive effect on bone health by effectively inhibiting osteoclast formation and TRAP activity, thereby interfering with bone resorption.
또한 상기 표에서 알 수 있듯이, 메틸설포닐메탄 100중량부, 칼슘 5~30중량부 및 로즈마리 추출물 2~15중량부를 사용하는 경우, 골다공증 예방효과가 극대화될 수 있다. In addition, as can be seen from the above table, when 100 parts by weight of methylsulfonylmethane, 5 to 30 parts by weight of calcium, and 2 to 15 parts by weight of rosemary extract are used, the osteoporosis prevention effect can be maximized.
아울러 메틸설포닐메탄 100중량부, 칼슘 5~30중량부, 로즈마리 추출물 2~15중량부 및 프로폴리스 추출물 2~10중량부를 사용하는 경우, 골다공증 예방효과가 극대화될 수 있다. In addition, when 100 parts by weight of methylsulfonylmethane, 5-30 parts by weight of calcium, 2-15 parts by weight of rosemary extract and 2-10 parts by weight of propolis extract are used, the osteoporosis prevention effect can be maximized.
또한 로즈마리 추출물 및 프로폴리스 추출물의 중량비가 60~80:20~40 인 경우, 골다공증 예방효과가 극대화될 수 있다. In addition, when the weight ratio of the rosemary extract and the propolis extract is 60 to 80: 20 to 40, the prevention effect of osteoporosis can be maximized.
(실시예 3) 실험동물의 체중 및 식이효율에 미치는 효과 (Example 3) Effect on body weight and dietary efficiency of experimental animals
난소 절제(ovariectomized)와 가짜수술(sham operation)을 실행한 생후 7주령의 암컷 SD Rat를 (주)중앙실험동물로부터 구입하여 고형식이를 먹이면서 14일간 환경에 적응시킨 후 체중에 따라 총 7군으로 분리하였다 (n=8 per group). Seven-week-old female SD Rat, which had been subjected to ovariectomized and sham operation, was purchased from Central Experimental Animal Co., Ltd. and acclimatized to the environment for 14 days while fed solid food, followed by a total of 7 according to body weight. They were divided into groups (n=8 per group).
실험동물에게 고형식이를 10주간 공급하였으며, 양성대조군으로 기제품인 호관원 프리미엄(MSM 25 mg/kg, calcium 4.5 mg/kg)을 선정하였으며, 비교를 위하여 로즈마리 추출물 저농도군(75 mg/kg) 및 고농도군(150 mg/kg)을 선정하였다. Solid food was supplied to the experimental animals for 10 weeks, and Hogwanwon Premium (MSM 25 mg/kg, calcium 4.5 mg/kg) was selected as a positive control group. For comparison, a low concentration group of rosemary extract (75 mg/kg) and a high concentration group (150 mg/kg) was selected.
최종적으로 MSM/Calcium/Rosemary 추출물을 혼합한 소재를 매일 경구투여로 정해진 양을 12주간 공급하였다. 사육장은 인공조명에 의하여 조명시간을 아침 9시부터 저녁 9시까지 12시간으로 조절하였으며 실내온도 25±3℃, 상대습도 50±10%로 유지하였다. 급수는 정수된 일반 식수를 사용하였고 사료와 급수는 제한하지 않았다. Finally, the material mixed with the MSM/Calcium/Rosemary extract was administered daily in a prescribed amount for 12 weeks. In the kennel, the lighting time was adjusted to 12 hours from 9 am to 9 pm by artificial lighting, and the room temperature was maintained at 25±3℃ and relative humidity of 50±10%. Purified general drinking water was used for water supply, and feed and water were not restricted.
1군: sham operation (가짜 수술군)Group 1: sham operation (sham operation group)
2군: 난소절제Group 2: Ovariectomy
3군: 난소절제 + 호관원 프리미엄 formula (MSM/칼슘)Group 3: Ovariectomy + Hogwanwon premium formula (MSM/calcium)
4군: 난소절제 + 로즈마리 저농도Group 4: Ovariectomy + Rosemary Low Concentration
5군: 난소절제 + 로즈마리 고농도Group 5: Ovariectomy + Rosemary High Concentration
6군: 난소절제 + MSM/칼슘/로즈마리 복합소재 formula (로즈마리 저농도)Group 6: Ovariectomy + MSM/calcium/rosemary composite formula (low concentration of rosemary)
7군: 난소절제 + MSM/칼슘/로즈마리 복합소재 formula (로즈마리 고농도)Group 7: Ovariectomy + MSM/calcium/rosemary composite formula (high concentration of rosemary)
MSM/Calcium/로즈마리 추출물 복합소재가 실험동물의 체중에 미치는 영향은 도 1에 제시된다. The effect of the MSM/Calcium/Rosemary extract composite material on the body weight of experimental animals is presented in FIG. 1 .
12주간의 실험기간동안 식이섭취효율은 유의적인 차이가 없었으나, 체중 증가는 1군(Sham)과 2군(OVA)간에 유의적인 차이가 나타났으며, 나머지 투여군에서는 유의적인 차이가 없었다. There was no significant difference in dietary intake efficiency during the 12-week experimental period, but there was a significant difference in weight gain between group 1 (Sham) and group 2 (OVA), and there was no significant difference in the other administration groups.
이는 난소절제에 의해 지방 축적률이 증가하여 체중증가가 나타났으며, MSM/Calcium/로즈마리 추출물 복합소재는 독성 및 섭취거부가 없으며 영양대사에 영향을 미치지 않는 것으로 나타났다. This resulted in weight gain due to increased fat accumulation by ovariectomy, and the MSM/Calcium/Rosemary extract composite material had no toxicity or refusal to consume, and did not affect nutritional metabolism.
(실시예 4) 실험동물의 신장, 간, 자궁 및 대퇴부 뼈에 미치는 효과 (Example 4) Effect on kidney, liver, uterus and femur bones of experimental animals
실험이 종료되는 시점(12주)에 실험동물을 희생시켜 신장, 간 및 자궁을 적출하여 분석하였으며, 좌측 및 우측 대퇴부 뼈를 적출하여 -70℃에서 보관하여 분석에 사용하였다. At the end of the experiment (12 weeks), the experimental animals were sacrificed and the kidneys, livers and uterus were extracted and analyzed.
대퇴부 뼈의 골밀도(bone mineral density, BMD)와 뼈 무기질 함량(bone mineral content, BMC)은 PIXmus (GE medical systems Lunar, USA)를 이용하여 측정하였다. 대퇴부 뼈 조직의 BMD는 cm2 당 g 단위로 표시하였으며, BMC는 g 단위로 표시하였다.Bone mineral density (BMD) and bone mineral content (BMC) of femoral bones were measured using PIXmus (GE medical systems Lunar, USA). BMD of femoral bone tissue was expressed in g per cm 2 , and BMC was expressed in g.
본 발명의 조성물이 실험동물의 대퇴부 뼈(BMC, BMD)에 미치는 영향은 도 2 및 3에 제시된다. The effect of the composition of the present invention on the femoral bones (BMC, BMD) of experimental animals is shown in FIGS. 2 and 3 .
대퇴부 뼈의 골밀도와 무기질 함량은 2군(OVA)이 1군(Sham)에 비해 감소하였으며, 5군 및 7군은 2군(OVA)에 비해 골밀도와 무기질 함량이 유의적으로 증가한 것으로 나타났다.
BMD and mineral content of femoral bone were decreased in group 2 (OVA) compared to group 1 (Sham), and
로즈마리 추출물 및 프로폴리스 추출물은 파골세포의 형성을 효과적으로 억제시켜 뼈의 흡수를 방해함으로써 뼈 건강에 긍정적 효과를 나타낼 수 있을 것으로 판단된다. Rosemary extract and propolis extract effectively inhibit the formation of osteoclasts and thus interfere with bone resorption, which is believed to have a positive effect on bone health.
Claims (5)
A pharmaceutical composition for preventing or treating osteoporosis comprising methylsulfonylmethane, calcium and rosemary extract as active ingredients.
상기 로즈마리 추출물은 로즈마리를 용매로 가열 추출하고 감압 농축하여 수득하는 것을 특징으로 하는 골다공증 예방 또는 치료용 약학 조성물.
According to claim 1,
The rosemary extract is a pharmaceutical composition for preventing or treating osteoporosis, characterized in that obtained by heating and extracting rosemary with a solvent and concentrating under reduced pressure.
상기 로즈마리 추출물은 로즈마리를 용매로 가열 추출하고 감압 농축하여 용매를 제거한 후, 정제수로 수용화한 수용성 추출물인 것을 특징으로 하는 골다공증 예방 또는 치료용 약학 조성물.
According to claim 1,
The rosemary extract is a pharmaceutical composition for preventing or treating osteoporosis, characterized in that it is a water-soluble extract obtained by heat-extracting rosemary with a solvent and concentration under reduced pressure to remove the solvent, and then solubilized with purified water.
상기 조성물은 메틸설포닐메탄 100중량부에 대하여 칼슘 5~30중량부 및 로즈마리 추출물 2~15중량부를 포함하는 것을 특징으로 하는 골다공증 예방 또는 치료용 약학 조성물.
According to claim 1,
The composition is a pharmaceutical composition for preventing or treating osteoporosis, characterized in that it comprises 5 to 30 parts by weight of calcium and 2 to 15 parts by weight of rosemary extract based on 100 parts by weight of methylsulfonylmethane.
A food composition for preventing or improving osteoporosis comprising methylsulfonylmethane, calcium and rosemary extract as active ingredients.
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