KR102592892B1 - Composition for preventing or treating diabetes comprising desoxycorticosterone glucoside - Google Patents

Composition for preventing or treating diabetes comprising desoxycorticosterone glucoside Download PDF

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KR102592892B1
KR102592892B1 KR1020210032538A KR20210032538A KR102592892B1 KR 102592892 B1 KR102592892 B1 KR 102592892B1 KR 1020210032538 A KR1020210032538 A KR 1020210032538A KR 20210032538 A KR20210032538 A KR 20210032538A KR 102592892 B1 KR102592892 B1 KR 102592892B1
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desoxycorticosterone
glucoside
present
diabetes
preventing
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KR20220128017A (en
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홍의주
이상렬
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충남대학교산학협력단
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7028—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages
    • A61K31/7034—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin
    • A61K31/704—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin attached to a condensed carbocyclic ring system, e.g. sennosides, thiocolchicosides, escin, daunorubicin
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/125—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives containing carbohydrate syrups; containing sugars; containing sugar alcohols; containing starch hydrolysates
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
    • A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2200/00—Function of food ingredients
    • A23V2200/30—Foods, ingredients or supplements having a functional effect on health
    • A23V2200/328—Foods, ingredients or supplements having a functional effect on health having effect on glycaemic control and diabetes

Abstract

본 발명은 데속시코르티코스테론 글루코사이드(desoxycorticosterone glucoside)를 포함하는 조성물 및 이의 당뇨병 예방 또는 치료 용도에 관한 것으로, 바람직하게는 제2형 당뇨병 또는 인슐린 저항성 예방, 개선 또는 치료 용도에 관한 것이다. The present invention relates to a composition containing desoxycorticosterone glucoside and its use for preventing or treating diabetes, preferably for preventing, improving or treating type 2 diabetes or insulin resistance.

Description

데속시코르티코스테론 글루코사이드를 포함하는 당뇨병 예방 또는 치료용 조성물 {Composition for preventing or treating diabetes comprising desoxycorticosterone glucoside}Composition for preventing or treating diabetes comprising desoxycorticosterone glucoside}

본 발명은 제2형 당뇨병 예방 또는 치료 효과가 우수한 데속시코르티코스테론 글루코사이드(desoxycorticosterone glucoside)를 유효성분으로 포함하는 조성물에 관한 것이다. The present invention relates to a composition containing desoxycorticosterone glucoside as an active ingredient, which has excellent effects in preventing or treating type 2 diabetes.

제 2 형 당뇨병 (T2D)은 전 세계적으로 발생하고 있으며 유병률은 수십 년에 걸쳐 증가 할 것으로 예상된다 (Wild et al., 2004). 제 2 형 당뇨병의 유병률은 아시아에서 특히 급증하고 있으며 그 발병원인은 라이프 스타일 의존성인 것으로 좌식생화 및 비만을 초래하는 식단에 기인한다 (Zheng et al., 2018). 제 2 형 당뇨 환자는 초기에 근육에서 인슐린 저항성을 보이고 나서 지방과 간에서 인슐린 저항성을 나타낸다 (Fernandez et al., 2001). 일반적으로 인슐린 매개 포도당 수송체 4 (GLUT4) 전위 (translocation)가 근육에서 포도당 이용을 촉진하지만, 제 2 형 당뇨병 기전은 인슐린 수용체 (insulin receptor; IR)의 근육 특이적인 knockout에 의해 유발되지 않음으로서 세포 내 신호 전달이 혈당 조절에 중요하다는 것을 시사한다 (Bruning et al., 1998; Zisman et al., 2000 ).Type 2 diabetes (T2D) is occurring worldwide and its prevalence is expected to increase over the decades (Wild et al., 2004). The prevalence of type 2 diabetes is rapidly increasing, especially in Asia, and its cause is lifestyle-dependent, resulting from diets that lead to sedentary living and obesity (Zheng et al., 2018). Type 2 diabetic patients initially exhibit insulin resistance in muscle and then in fat and liver (Fernandez et al., 2001). Although insulin-mediated glucose transporter 4 (GLUT4) translocation generally promotes glucose utilization in muscle, the mechanism of type 2 diabetes is not triggered by muscle-specific knockout of the insulin receptor (IR) in the cells. This suggests that intracellular signaling is important for glycemic control (Bruning et al., 1998; Zisman et al., 2000).

프로게스테론 수용체 막 성분 1 (Pgrmc1)은 전형적인 프로게스테론 수용체와는 달리 직접적인 유전자 조절 전사적 효과를 갖지 않는 세포 표면의 새로운 프로게스테론 결합 수용체이다 (Kasubuchi et al., 2017). 이전 연구에서 PGRMC1은 콜레스테롤 합성에 기여하고 (Hughes et al., 2007; Rohe et al., 2009), 콜레스테롤 항상성의 핵심 조절자인 INSIG-1에 결합하는 것으로 알려져 있다 (Suchanek et al., 2005). 또한, PGRMC1은 췌장 β 세포에서 순환 AMP (cAMP) 축적에 관여하는 글루카곤 유사 펩타이드-1 수용체 (GLP-1R)의 기능적 성분으로 확인되었다 (Zhang et al., 2014). 최근 연구에 따르면 Pgrmc1이 고지방 식단에 유발된 지방간을 억제하는 것이 관찰되었다 (Lee et al., 2018). Pgrmc1의 내분비 관련성이 나타나고 있지만 Pgrmc1이 제 2 형 당뇨병과 관련이 있는지 여부에 대해 풀어야 할 과제가 존재한다.Progesterone receptor membrane component 1 (Pgrmc1) is a novel progesterone-binding receptor on the cell surface that, unlike typical progesterone receptors, does not have direct gene regulatory transcriptional effects (Kasubuchi et al., 2017). Previous studies have shown that PGRMC1 contributes to cholesterol synthesis (Hughes et al., 2007; Rohe et al., 2009) and is known to bind to INSIG-1, a key regulator of cholesterol homeostasis (Suchanek et al., 2005). Additionally, PGRMC1 was identified as a functional component of the glucagon-like peptide-1 receptor (GLP-1R), which is involved in cyclic AMP (cAMP) accumulation in pancreatic β cells (Zhang et al., 2014). A recent study observed that Pgrmc1 suppresses fatty liver disease induced by a high-fat diet (Lee et al., 2018). Although the endocrine relevance of Pgrmc1 is emerging, whether Pgrmc1 is associated with type 2 diabetes remains to be resolved.

이전 연구에 따르면 PGRMC1과 인슐린 수용체 베타 (IRβ) 간의 단백질 상호 작용이 확인되었으며 Pgrmc1 하위 조절은 인슐린 치료 없이 포도당 흡수를 감소 시킨 것을 세포주를 통해 확인하였다. 하지만 IRβ-GLUT4 신호 전달의 중심 조절인 인산화된 AKT의 수준이 유의적으로 증가하였다 (Hampton et al., 2018). 세포막 IRβ 및 인산화된-AKT 수준이 Pgrmc1 조절에서 모순되는 결과를 보여주기 때문에, 본 발명에서는 Pgrmc1이 AKT 인산화를 활성화하는 세포 내 신호 전달에 연루되어 있다고 예상하였다. PGRMC1 수준을 조절하는 내인성 호르몬인 데속시코르티코스테론 글루코사이드(desoxycorticosterone glucoside)를 사용하여 제2형 당뇨병에서 PGRMC1의 역할과 새로운 치료 방법에 사용될 가능성을 탐구하였다.According to a previous study, protein interaction between PGRMC1 and insulin receptor beta (IRβ) was confirmed, and Pgrmc1 downregulation decreased glucose uptake without insulin treatment through cell lines. However, the level of phosphorylated AKT, a central regulator of IRβ-GLUT4 signaling, was significantly increased (Hampton et al., 2018). Because cell membrane IRβ and phosphorylated-AKT levels show contradictory results in Pgrmc1 regulation, we predicted that Pgrmc1 is involved in intracellular signaling that activates AKT phosphorylation. We explored the role of PGRMC1 in type 2 diabetes and its potential use in new treatment methods using desoxycorticosterone glucoside, an endogenous hormone that regulates PGRMC1 levels.

한국공개특허 제2010-0094495호Korean Patent Publication No. 2010-0094495 한국공개특허 제2008-0063401호Korean Patent Publication No. 2008-0063401

본 발명은 상기와 같은 요구에 의해 도출된 것으로서, 본 발명자들은 데속시코르티코스테론 글루코사이드를 포함하는 조성물의 당뇨병 예방 또는 개선, 특히 제2형 당뇨병 또는 인슐린 저항성 개선 효과를 확인하여 본 발명을 완성하였다. The present invention was developed in response to the above-mentioned needs, and the present inventors completed the present invention by confirming the effect of a composition containing desoxycorticosterone glucoside on preventing or improving diabetes, especially type 2 diabetes or improving insulin resistance. .

상기 과제를 해결하기 위하여, 본 발명은 데속시코르티코스테론 글루코사이드(desoxycorticosterone glucoside)를 유효성분으로 포함하는 당뇨병 또는 이의 합병증 예방 또는 치료용 약학 조성물을 제공한다. In order to solve the above problems, the present invention provides a pharmaceutical composition for preventing or treating diabetes or its complications containing desoxycorticosterone glucoside as an active ingredient.

본 발명의 일 예에서, 당뇨병은 제2형 당뇨병인 것을 특징으로 하는 것이다.In one example of the present invention, diabetes is characterized as type 2 diabetes.

본 발명의 일 예에서, 데속시코르티코스테론 글루코사이드는 PGRMC1 억제와 당 분해를 유도하는 것을 특징으로 한다.In one example of the present invention, desoxycorticosterone glucoside is characterized by inhibiting PGRMC1 and inducing glycolysis.

본 발명의 다른 예에서, 상기 합병증은 당뇨병성 심장병, 당뇨병성 신장장애, 당뇨병성 혈관장애 또는 당뇨병성 신경장애인 것을 특징으로 한다.In another example of the present invention, the complication is characterized by diabetic heart disease, diabetic kidney disease, diabetic vascular disease, or diabetic neuropathy.

또한, 본 발명은 데속시코르티코스테론 글루코사이드(desoxycorticosterone glucoside)를 유효성분으로 포함하는 당뇨병 또는 이의 합병증 예방 또는 개선용 식품 조성물에 관한 것이다. Additionally, the present invention relates to a food composition for preventing or improving diabetes or its complications containing desoxycorticosterone glucoside as an active ingredient.

본 발명의 다른 예에서, 본 발명은 데속시코르티코스테론 글루코사이드(desoxycorticosterone glucoside)를 유효성분으로 포함하는 인슐린 저항성 예방 또는 치료용 약학 조성물에 관한 것이다. In another example of the present invention, the present invention relates to a pharmaceutical composition for preventing or treating insulin resistance containing desoxycorticosterone glucoside as an active ingredient.

본 발명의 또 다른 예에서, 데속시코르티코스테론 글루코사이드(desoxycorticosterone glucoside)를 유효성분으로 포함하는 인슐린 저항성 예방 또는 개선용 식품에 관한 것이다. In another example, the present invention relates to a food for preventing or improving insulin resistance containing desoxycorticosterone glucoside as an active ingredient.

본 발명의 데속시코르티코스테론 글루코사이드를 포함하는 조성물은 당 대사와 PGRMC1에 대한 효과가 있으므로 당뇨병 예방 또는 치료에 유용하게 사용될 수 있다. The composition containing desoxycorticosterone glucoside of the present invention has an effect on glucose metabolism and PGRMC1 and can therefore be usefully used in the prevention or treatment of diabetes.

도 1은 본 발명의 일 구현 예에 따른 데속시코르티코스테론 글루코사이드 처리 A204 근육세포에서 PGRMC1의 발현에 대한 효과를 나타낸 것이다.
도 2는 본 발명의 일 구현 예에 따른 데속시코르티코스테론 글루코사이드 처리 인슐린 처리 A204 근육세포에서 세포외 산성화율(ECAR, extracellular acidification rate)에 대한 효과를 나타낸 것이다.
도 3은 본 발명의 일 구현 예에 따른 데속시코르티코스테론 아세테이트 처리 인슐린 처리 A204 근육세포에서 세포외 산성화율(ECAR, extracellular acidification rate)에 대한 효과를 나타낸 것이다.Figure 1 shows the effect on the expression of PGRMC1 in A204 muscle cells treated with desoxycorticosterone glucoside according to an embodiment of the present invention.
Figure 2 shows the effect on extracellular acidification rate (ECAR) in desoxycorticosterone glucoside-treated and insulin-treated A204 muscle cells according to an embodiment of the present invention.
Figure 3 shows the effect on extracellular acidification rate (ECAR) in desoxycorticosterone acetate-treated, insulin-treated A204 muscle cells according to an embodiment of the present invention.

이하, 본 발명의 바람직한 구현예에 대하여 상세히 설명한다. 또한, 하기의 설명에서는 구체적인 구성요소 등과 같은 많은 특정 사항들이 도시되어 있는데, 이는 본 발명의 보다 전반적인 이해를 돕기 위해서 제공된 것일 뿐 이러한 특정 사항들 없이도 본 발명이 실시될 수 있음은 이 기술분야에서 통상의 지식을 가진 자에게는 자명하다 할 것이다. 그리고, 본 발명을 설명함에 있어서, 관련된 공지 기능 혹은 구성에 대한 구체적인 설명이 본 발명의 요지를 불필요하게 흐릴 수 있다고 판단되는 경우 그 상세한 설명을 생략한다.Hereinafter, preferred embodiments of the present invention will be described in detail. In addition, in the following description, many specific details, such as specific components, are shown, but this is provided to facilitate a more general understanding of the present invention, and it is known in the art that the present invention can be practiced without these specific details. It will be self-evident to those who have the knowledge. Additionally, in describing the present invention, if it is determined that a detailed description of a related known function or configuration may unnecessarily obscure the gist of the present invention, the detailed description will be omitted.

본 발명에서 용어, "예방" 또는 "방지"라 함은 질환의 원인으로부터 발생을 억제하거나 지연시키는 것을 의미한다.In the present invention, the term “prevention” or “prevention” means suppressing or delaying the occurrence of a disease from its cause.

본 명세서에서, "치료"라 함은 완전히 치유하지 않아도 증상의 진전 및/또는 악화를 억제하여 손상의 진행을 멈추거나, 또는 증상의 일부 혹은 전부를 개선하여 치유의 방향으로 유도하는 것을 의미한다.In this specification, “treatment” means stopping the progression of damage by suppressing the progression and/or worsening of symptoms even if not completely cured, or improving some or all of the symptoms and leading toward healing.

본 발명에서 용어 “개선”은 증상이 호전 또는 이롭게 변경되는 모든 행위를 의미한다.In the present invention, the term “improvement” refers to any action that improves or beneficially changes symptoms.

본 발명의 목적을 달성하기 위하여, 본 발명은 데속시코르티코스테론 글루코사이드(desoxycorticosterone glucoside)를 유효성분으로 포함하는 당뇨병 또는 이의 합병증 예방 또는 치료용 약학 조성물을 제공한다.In order to achieve the object of the present invention, the present invention provides a pharmaceutical composition for preventing or treating diabetes or its complications containing desoxycorticosterone glucoside as an active ingredient.

본 발명에서, 데속시코르티코스테론 글루코사이드(desoxycorticosterone glucoside)는 11β-하이드록시프로게스테론의 에피머로서 11β-하이드록시스테로이드 디하이드로게네이즈의 아이소폼 1 및 2의 경쟁적 억제자로서 작용한다.In the present invention, desoxycorticosterone glucoside is an epimer of 11β-hydroxyprogesterone and acts as a competitive inhibitor of isoforms 1 and 2 of 11β-hydroxysteroid dehydrogenase.

상기 당뇨병은 제1형 당뇨병 또는 제2형 당뇨병에 관한 것이고, 바람직하게는 제2형 당뇨병에 관한 것이나, 이에 제한되는 것은 아니다.The diabetes relates to type 1 diabetes or type 2 diabetes, and preferably relates to type 2 diabetes, but is not limited thereto.

상기 합병증은 당뇨병으로 인해 발생하는 모든 합병증에 관한 것이고, 바람직하게는 당뇨병성 심장병, 당뇨병성 신장장애, 당뇨병성 혈관장애 또는 당뇨병성 신경장애에 관한 것이나, 이에 제한되는 것은 아니다.The complication relates to all complications arising from diabetes, and preferably relates to diabetic heart disease, diabetic kidney disease, diabetic vascular disease, or diabetic neuropathy, but is not limited thereto.

본 발명의 다른 예에서, 본 발명은 데속시코르티코스테론 글루코사이드(desoxycorticosterone glucoside)를 유효성분으로 포함하는 인슐린 저항성 예방 또는 치료용 약학 조성물에 관한 것이다. In another example of the present invention, the present invention relates to a pharmaceutical composition for preventing or treating insulin resistance containing desoxycorticosterone glucoside as an active ingredient.

본 발명의 일 예에서, 데속시코르티코스테론 글루코사이드는 PGRMC1 억제 및 당 분해를 유도하는 것을 특징으로 한다.In one embodiment of the present invention, desoxycorticosterone glucoside is characterized by inhibiting PGRMC1 and inducing glycolysis.

또한, 본 발명의 일 구현예에 따른 조성물에서, 상기 조성물은 당뇨병에 유용한 추가 성분을 포함할 수 있고, 추가성분은 화합물, 천연물을 포함하는 당뇨병에 효과가 공지된 모든 성분을 포함할 수 있다.Additionally, in the composition according to one embodiment of the present invention, the composition may include additional ingredients useful for diabetes, and the additional ingredients may include all ingredients known to be effective in diabetes, including compounds and natural products.

본 발명의 조성물은 조성물의 제조에 통상적으로 사용하는 적절한 담체, 부형제 및 희석제를 더 포함하여 모든 형태의 조성물로 제조될 수 있고, 바람직하게는 약학 조성물, 건강기능식품 조성물 또는 화장료 조성물의 형태로 제조될 수 있으나, 이에 제한되는 것은 아니다. The composition of the present invention can be prepared in any form by further including appropriate carriers, excipients, and diluents commonly used in the preparation of the composition, and is preferably prepared in the form of a pharmaceutical composition, health functional food composition, or cosmetic composition. It may be, but is not limited to this.

본 발명의 약학 조성물은, 통상의 방법에 따라 산제, 과립제, 정제, 캡슐제, 현탁액, 에멀젼, 시럽, 에어로졸 등의 경구형 제형, 외용제, 좌제 및 멸균 주사용액의 형태로 제제화될 수 있으며, 산제, 정제, 캡슐제, 주사제 및 액제가 보다 바람직하다. 이러한 제제화는 약제학 분야에서 통상적으로 행하여지는 방법으로 수행될 수 있으며, Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA에 개시되어 있는 방법을 이용하여 각 질환에 따라 또는 성분에 따라 바람직하게 제제화할 수 있다. The pharmaceutical composition of the present invention can be formulated in the form of oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories, and sterile injectable solutions according to conventional methods. , tablets, capsules, injections and liquid formulations are more preferable. This formulation can be performed by a method commonly performed in the pharmaceutical field, and can be preferably formulated according to each disease or ingredient using the method disclosed in Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA.

상기 약학 조성물에 포함될 수 있는 담체, 부형제 및 희석제로는 락토즈, 덱스트로즈, 수크로스, 솔비톨, 만니톨, 자일리톨, 에리스리톨, 말티톨, 전분, 아카시아 고무, 알지네이트, 젤라틴, 칼슘 포스페이트, 칼슘 실리케이트, 셀룰로오스, 메틸 셀룰로오스, 미정질 셀룰로오스, 폴리비닐피롤리돈, 물, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트, 탈크, 마그네슘 스테아레이트, 및 광물유 등을 포함한다.Carriers, excipients, and diluents that may be included in the pharmaceutical composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, and cellulose. , methyl cellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil.

제제화할 경우에는 보통 사용하는 충진제, 증량제, 결합제, 습윤제, 붕해제, 계면활성제 등의 희석제 또는 부형제를 추가로 사용하여 조제될 수 있다.When formulated, it can be prepared by additionally using diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants.

경구 투여를 위한 고형 제제에는 정제, 환제, 산제, 과립제, 캡슐제 등이 포함되며, 이러한 고형 제제는 적어도 하나 이상의 부형제, 예를 들면 전분, 칼슘카보네이트(calcium carbonate), 수크로오스(sucrose) 또는 락토오스(lactose), 젤라틴 등을 섞어 조제된다. 또한, 단순한 부형제 이외에 마그네슘 스테아레이트, 탈크 같은 윤활제들도 사용된다.Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid preparations contain at least one excipient, such as starch, calcium carbonate, sucrose, or lactose ( It is prepared by mixing lactose, gelatin, etc. Additionally, in addition to simple excipients, lubricants such as magnesium stearate and talc are also used.

경구 투여를 위한 액상 제제로는 현탁액, 내용액제, 유제, 시럽제 등이 해당되는데, 흔히 사용되는 단순 희석제인 물, 리퀴드 파라핀 이외에 여러 가지 부형제, 예를 들면 습윤제, 감미제, 방향제, 보존제 등이 포함될 수 있다. 비경구 투여를 위한 제제에는 멸균된 수용액, 비수성용제, 현탁제, 유제, 동결건조 제제, 좌제가 포함된다. 비수용제, 현탁제로는 프로필렌글리콜(propylene glycol), 폴리에틸렌 글리콜, 올리브 오일과 같은 식물성 기름, 에틸올레이트와 같은 주사 가능한 에스테르 등이 사용될 수 있다. 좌제의 기제로는 위텝솔(witepsol), 마크로골, 트윈(tween), 카카오지, 라우린지, 글리세로제라틴 등이 사용될 수 있다. Liquid preparations for oral administration include suspensions, oral solutions, emulsions, and syrups. In addition to the commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. there is. Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous agents and suspensions may include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate. As a base for suppositories, witepsol, macrogol, tween, cacao, laurin, glycerogeratin, etc. can be used.

본 발명의 약학 조성물의 바람직한 투여량은 환자의 상태 및 체중, 질병의 정도, 약물형태, 투여경로 및 기간에 따라 다르지만, 당업자에 의해 적절하게 선택될 수 있다. 그러나, 바람직한 효과를 위해서 본 발명의 약학 조성물은 1 일 0.01 내지 99.9 중량%, 바람직하게는 0.1 내지 99 중량%로 포함될 수 있다. 일일 투여량은 약 0.1 내지 1,000 mg/kg으로, 바람직하게는 100~300 mg/kg일 수 있다. The preferred dosage of the pharmaceutical composition of the present invention varies depending on the patient's condition and weight, degree of disease, drug form, administration route and period, but can be appropriately selected by a person skilled in the art. However, for desirable effects, the pharmaceutical composition of the present invention may be included in an amount of 0.01 to 99.9% by weight, preferably 0.1 to 99% by weight, per day. The daily dosage may be about 0.1 to 1,000 mg/kg, preferably 100 to 300 mg/kg.

또한, 본 발명은 데속시코르티코스테론 글루코사이드(desoxycorticosterone glucoside)를 유효성분으로 포함하는 당뇨병 또는 이의 합병증 예방 또는 개선용 식품 조성물에 관한 것이다. Additionally, the present invention relates to a food composition for preventing or improving diabetes or its complications containing desoxycorticosterone glucoside as an active ingredient.

본 발명의 또 다른 예에서, 데속시코르티코스테론 글루코사이드(desoxycorticosterone glucoside)를 유효성분으로 포함하는 인슐린 저항성 예방 또는 개선용 식품에 관한 것이다. In another example, the present invention relates to a food for preventing or improving insulin resistance containing desoxycorticosterone glucoside as an active ingredient.

상기 조성물을 첨가할 수 있는 건강기능식품으로는 예를 들어, 각종 일반식품류, 음료, 껌, 차, 비타민 복합제 등이 있다.Health functional foods to which the composition can be added include, for example, various general foods, beverages, gum, tea, vitamin complexes, etc.

또한, 상기 조성물은 질환의 예방 효과를 목적으로 식품 또는 음료에 첨가될 수 있다. 이때, 식품 또는 음료 중의 상기 추출물의 양은 전체 식품 중량의 0.01 내지 15 중량%로 가할 수 있으며, 건강 음료 조성물은 100 g을 기준으로 0.02 내지 5 g, 바람직하게는 0.3 내지 1 g의 비율로 가할 수 있다.Additionally, the composition can be added to food or beverages for the purpose of preventing diseases. At this time, the amount of the extract in the food or beverage can be added at 0.01 to 15% by weight of the total weight of the food, and the health drink composition can be added at a rate of 0.02 to 5 g, preferably 0.3 to 1 g, based on 100 g. there is.

본 발명의 건강기능음료 조성물은 지시된 비율로 필수 성분으로서 상기 추출물을 함유하는 것 외에 다른 성분에는 특별한 제한이 없으며, 통상의 음료와 같이 여러 가지 향미제 또는 천연 탄수화물 등의 추가 성분을 함유할 수 있다. 상술한 천연탄수화물의 예로는 모노사카라이드, 예를 들어 포도당, 과당 등; 디사카라이드, 예를들어 말토오스, 수크로오스 등; 및 폴리사카라이드, 예를 들어 덱스트린, 시클로덱스트린 등과 같은 통상적인 당, 및 자일리톨, 솔비톨, 에리트리톨 등의 당알콜이다. 상술한 것 이외의 향미제로서 천연 향미제인 타우마틴, 스테비아 추출물, 예를 들어 레바우디오시드 A, 글리시르히진 등; 및 합성 향미제, 예를 들어 사카린, 아스파르탐 등을 유리하게 사용할 수 있다. 상기 천연 탄수화물의 비율은 본 발명의 조성물 100 g당 일반적으로 약 1 내지 20 g, 바람직하게는 약 5 내지 12 g이다. 상기 외에 본 발명의 추출물은 여러 가지 영양제, 비타민, 광물(전해질), 합성 및 천연 풍미제, 착색제 및 중진제(치즈, 초콜릿 등), 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알콜, 탄산 음료에 사용되는 탄산화제 등을 함유할 수 있다. 그밖에 본 발명의 추출물들은 천연 과일 쥬스, 과일 쥬스 음료 및 야채 음료의 제조를 위한 과육을 함유할 수 있다. 이러한 성분은 독립적으로 또는 조합하여 사용할 수 있다. 이때, 첨가제의 비율은 그다지 중요하지는 않지만 본 발명의 조성물 100 중량부 당 0.01 내지 약 20 중량부의 범위에서 선택되는 것이 일반적이다.The health functional drink composition of the present invention has no particular restrictions on other ingredients other than containing the extract as an essential ingredient in the indicated ratio, and may contain additional ingredients such as various flavoring agents or natural carbohydrates like a conventional beverage. there is. Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose, etc.; Disaccharides such as maltose, sucrose, etc.; and polysaccharides, such as common sugars such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. Flavoring agents other than those mentioned above include natural flavoring agent thaumatin, stevia extract, such as rebaudioside A, glycyrrhizin, etc.; and synthetic flavoring agents such as saccharin, aspartame, etc. can be advantageously used. The proportion of natural carbohydrates is generally about 1 to 20 g, preferably about 5 to 12 g, per 100 g of the composition of the present invention. In addition to the above, the extract of the present invention contains various nutrients, vitamins, minerals (electrolytes), synthetic and natural flavoring agents, colorants and thickening agents (cheese, chocolate, etc.), pectic acid and its salts, alginic acid and its salts, organic acids, and protective properties. It may contain colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc. In addition, the extracts of the present invention may contain pulp for the production of natural fruit juice, fruit juice beverages, and vegetable beverages. These ingredients can be used independently or in combination. At this time, the ratio of the additive is not very important, but is generally selected in the range of 0.01 to about 20 parts by weight per 100 parts by weight of the composition of the present invention.

본 발명의 이점 및 특징, 그리고 그것들을 달성하는 방법은 상세하게 후술되어있는 실시예들을 참조하면 명확해질 것이다. 그러나 본 발명은 이하에서 개시되는 실시예들에 한정되는 것이 아니라 서로 다른 다양한 형태로 구현될 것이며, 단지 본 실시예들은 본 발명의 개시가 완전하도록 하고, 본 발명이 속하는 기술 분야에서 통상의 지식을 가진 자에게 발명의 범주를 완전하게 알려주기 위해 제공되는 것이며, 본 발명은 청구항의 범주에 의해 정의될 뿐이다.The advantages and features of the present invention and methods for achieving them will become clear with reference to the embodiments described in detail below. However, the present invention is not limited to the embodiments disclosed below and will be implemented in various different forms. The present embodiments are merely provided to ensure that the disclosure of the present invention is complete and that common knowledge in the technical field to which the present invention pertains is not limited. It is provided to fully inform those who have the scope of the invention, and the present invention is only defined by the scope of the claims.

<실시예> <Example>

1. 화합물1. Compound

Insulin (I0908, Sigma), glucose (G8270, Sigma), deoxycorticosterone acetate (D0047, TCI) 및 desoxycorticosterone glucoside를 사용하였다.Insulin (I0908, Sigma), glucose (G8270, Sigma), deoxycorticosterone acetate (D0047, TCI), and desoxycorticosterone glucoside were used.

2. 웨스턴 블롯팅2. Western blotting

단백질 샘플은 조직 및 세포의 균질환, 원심 분리 및 초음파 처리를 포함하여 순차적 절차로 획득하였다. 단백질 샘플의 농도는 PRO-Measure solution (Intron, #21011)로 측정하였고 동일한 양의 단백질을 로드하여 SDS-PAGE로 분리하였다. 겔은 막에 블롯팅하였고, 막은 블로킹한 후 1차 항체로 반응하였다. 다음날 막을 2차 항체로 반응시켰다. 결과는 ECL solution (XLS025-0000, Cyanagen)과 Chemi Doc (Fusion Solo, Vilber Lourmat)으로 검출하였다. Protein samples were obtained by sequential procedures including disaggregation, centrifugation, and sonication of tissues and cells. The concentration of the protein sample was measured using PRO-Measure solution (Intron, #21011), and the same amount of protein was loaded and separated by SDS-PAGE. The gel was blotted onto a membrane, and the membrane was blocked and then reacted with primary antibody. The next day, the membrane was reacted with secondary antibody. The results were detected with ECL solution (XLS025-0000, Cyanagen) and Chemi Doc (Fusion Solo, Vilber Lourmat).

1차 항체는 β-ACTIN (Santa Cruz, sc-130656) 및 PGRMC1 (CST, #13856)를 사용하였다.Primary antibodies used were β-ACTIN (Santa Cruz, sc-130656) and PGRMC1 (CST, #13856).

2차 항체는 Goat anti-rabbit IgG HRP (Thermo Fisher, 31460)를 사용하였다. Goat anti-rabbit IgG HRP (Thermo Fisher, 31460) was used as the secondary antibody.

3. 세포 배양3. Cell culture

모든 세포 배양 시약은 Welgene (Gyungsan, Korea)에서 구입하였다. A204 사람 횡문근육종 세포를 5% FBS, 페니실린(100 U/mol) 및 스트렙토마이신(100 μg/ml)로 보충한 McCoy 배양액 (Welgene, LM005-01)을 사용하여 5% CO2 대기중에서 37℃ 로 배양하였다. 세포는 혈청이 배제된 McCoy 배지로 5 시간 동안 배양하여 인슐린을 제거하였다. All cell culture reagents were purchased from Welgene (Gyungsan, Korea). A204 human rhabdomyosarcoma cells were cultured at 37°C in a 5% CO2 atmosphere using McCoy medium (Welgene, LM005-01) supplemented with 5% FBS, penicillin (100 U/mol), and streptomycin (100 μg/ml). did. Cells were cultured in serum-excluded McCoy medium for 5 hours to remove insulin.

4. 당분해 스트레스 테스트4. Glycolytic stress test

A204 세포는 감염되어 seahorse cell culture plate에서 분리되었다. 세포는 혈청이 제외된 McCoy 배지(glucose 4500 mg/L)로 5시간 동안 배양하여 내재적 호르몬을 제거하고, 저 포도당 혈청 배제 McCoy 배지(glucose 500 mg/L)에서 1시간 동안 배양하였다. 글루타민, 소듐 피루베이트 및 글루코스 동량을 포함하는 XFp 배지(103575-100, Agilent technologies)에서 40 min 내지 1 시간 동안 탈카르복시화하였고, 세포를 배양하였다. 당분해 스트레스 테스트를 위하여 인슐린 (100 nM) 및 글루코스(25 mM)를 사용하였고 extracellular acidification rate (ECAR) 를 측정하였다. Seahorse XFp analyzer (Agilent technologies) 및 Seahorse XFp, XFp FluxPak (103022-100, Agilent technologies)를 사용하였다. A204 cells were infected and isolated from seahorse cell culture plates. Cells were cultured in serum-excluded McCoy medium (glucose 4500 mg/L) for 5 hours to remove endogenous hormones, and then cultured in low-glucose serum-excluded McCoy medium (glucose 500 mg/L) for 1 hour. Decarboxylation was performed for 40 min to 1 hour in XFp medium (103575-100, Agilent technologies) containing equal amounts of glutamine, sodium pyruvate, and glucose, and the cells were cultured. For the glycolytic stress test, insulin (100 nM) and glucose (25 mM) were used, and extracellular acidification rate (ECAR) was measured. Seahorse XFp analyzer (Agilent technologies) and Seahorse XFp, XFp FluxPak (103022-100, Agilent technologies) were used.

7. 통계 분석7. Statistical analysis

결과는 평균±표준편차로 나타내었다. 평균의 차이는 스튜던트 t-테스트로 얻었고, Graph Pad Software (GraphPad Inc., San Diego, CA)로 수행하였다. Results were expressed as mean ± standard deviation. Differences in means were obtained by Student's t-test and performed with Graph Pad Software (GraphPad Inc., San Diego, CA).

<시험예> <Test example>

1. Desoxycorticosterone glucoside 처리 A204 근육세포에서 PGRMC1 발현.1. PGRMC1 expression in desoxycorticosterone glucoside treated A204 muscle cells.

Desoxycorticosterone glucoside 처리된 세포를 웨스턴 블롯팅 방법으로 분석하였을 때, desoxycorticosterone glucoside 처리된 세포에서 PGRMC1 단백질 수준이 감소 (p<0.05, 50nM과 100nM 에 각각 69.3 및 67%) 된 것을 확인하였다. When cells treated with desoxycorticosterone glucoside were analyzed by Western blotting, it was confirmed that the PGRMC1 protein level was decreased (p<0.05, 69.3 and 67% at 50nM and 100nM, respectively) in cells treated with desoxycorticosterone glucoside.

2. Desoxycorticosterone glucoside 처리 A204 근육세포에서 ECAR2. ECAR in desoxycorticosterone glucoside treated A204 muscle cells

Desoxycorticosterone glucoside 처리된 세포를 실시간 대사측정 방법으로 분석하였을 때, desoxycorticosterone glucoside 처리된 세포에서 포도당 노출 후 세포 외 산성화 율 (ECAR, extracellular acidification rate)이 증가되는 것을 (p<0.05, 1.64-fold) 확인하였다. When desoxycorticosterone glucoside-treated cells were analyzed using a real-time metabolic measurement method, it was confirmed that the extracellular acidification rate (ECAR) increased (p<0.05, 1.64-fold) after glucose exposure in desoxycorticosterone glucoside-treated cells. .

3. Desoxycorticosterone acetate 처리 A204 근육세포에서 ECAR3. ECAR in desoxycorticosterone acetate treated A204 muscle cells

Deoxycorticosterone acetate 처리된 세포를 실시간 대사측정 방법으로 분석하였을 때, deoxycorticosterone acetate 처리된 세포에서 포도당 노출 후 세포 외 산성화 율이 증가되지 않는 것을 확인하였다.When deoxycorticosterone acetate-treated cells were analyzed using a real-time metabolic measurement method, it was confirmed that the extracellular acidification rate did not increase in deoxycorticosterone acetate-treated cells after exposure to glucose.

ReferenceReference

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Claims (12)

데속시코르티코스테론 글루코사이드(desoxycorticosterone glucoside)를 유효성분으로 포함하는 제2형 당뇨병 예방 또는 치료용 약학 조성물Pharmaceutical composition for preventing or treating type 2 diabetes containing desoxycorticosterone glucoside as an active ingredient 삭제delete 제1항에 있어서, 데속시코르티코스테론 글루코사이드는 PGRMC1 억제 및 당 분해를 유도하는 것을 특징으로 하는 제2형 당뇨병 예방 또는 치료용 약학 조성물The pharmaceutical composition for preventing or treating type 2 diabetes according to claim 1, wherein desoxycorticosterone glucoside inhibits PGRMC1 and induces glycolysis. 삭제delete 데속시코르티코스테론 글루코사이드(desoxycorticosterone glucoside)를 유효성분으로 포함하는 제2형 당뇨병 예방 또는 개선용 식품 조성물Food composition for preventing or improving type 2 diabetes containing desoxycorticosterone glucoside as an active ingredient 삭제delete 제5항에 있어서, 데속시코르티코스테론 글루코사이드는 PGRMC1 억제 및 당 분해를 유도하는 것을 특징으로 하는 제2형 당뇨병 예방 또는 개선용 식품 조성물
The food composition for preventing or improving type 2 diabetes according to claim 5, wherein desoxycorticosterone glucoside inhibits PGRMC1 and induces glycolysis.
 삭제delete 삭제delete 삭제delete 삭제delete 삭제delete
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