KR20130076579A - Betulinic acid and its derivatives with anti-hyperglycemic activity and compositions comprising the same - Google Patents

Abstract

PURPOSE: Betulinic acid and a derivative thereof are provided to induce anti-diabetic activity through intraperitoneal administration or oral administration, to be used as a functional food or a food additive, and to be formulated in a composition for oral administration and injection. CONSTITUTION: Betulinic acid of chemical formulas 1-3 or a derivative thereof has anti-diabetic activity. A pharmaceutical composition for preventing or treating diabetes contains betulinic acid or the derivative thereof as an active ingredient. A functional health food composition for treating diabetes contains betulinic acid or the derivative thereof as an active ingredient. A pharmaceutical composition contains betulinic acid and other carrier or excipient, and is provided in the form of a tablet, a pill, a granule, a liquid, and a capsule.

Description

Betulinic acid and its derivatives having antidiabetic activity and compositions comprising same {Betulinic acid and its derivatives with anti-hyperglycemic activity and compositions comprising the same}

The present invention relates to betulinic acid (Betulinic acid: BA) having an antidiabetic activity, a derivative thereof, and a composition comprising the same, and more particularly, to reveal the antidiabetic activity of betulinic acid and its derivatives, It relates to an antidiabetic pharmaceutical composition and functional health food composition.

In modern society, the incidence rate of various diseases is increasing due to environmental pollution and the increase of the elderly population. Among them, adult diseases and degenerative diseases that occur mainly in the mid 40s show a very high trend. In Korea, cancer occurring in various parts of a single disease accounts for the highest mortality rate, followed by cardiovascular disease and stroke, hyperlipidemia, hypertension, arteriosclerosis, diabetes, and degenerative arthritis. On the other hand, interest in personal health continues to increase as income levels and living standards increase due to economic development.In particular, health maintenance laws, health foods and supplements, and preventive medicines are spotlighted for prevention and treatment in relation to adult diseases. Is getting. Among these diseases, hypertension, arteriosclerosis, heart disease, stroke, etc. are representative circulatory diseases, but there are no drugs that guarantee cure even though many therapeutic drugs have been developed. In addition, these cardiovascular diseases, including diabetes, are often life-threatening with complications rather than direct causes of death. Therefore, maintaining the health after the middle age is an important factor for preventing these diseases. have.

Diabetes mellitus is a disease caused by the lack of insulin secreted by the pancreatic beta cells or abnormal action, causing blood sugar levels to rise and nutrients not to be used smoothly. Blood sugar is excreted in the urine or converted into cholesterol or triglycerides in the blood. to be. Diabetes does not produce as much insulin as the body needs, or the produced insulin does not work properly in the body, causing blood sugar levels to rise, causing excess sugar to be excreted in the urine, resulting in loss of energy and fatigue. do. It is known that about 5% of Korean adults are diabetic patients. Diabetes mellitus causes complications such as diabetic retinopathy or diabetic nephropathy, vascular disorders, neurological disorders and arteriosclerosis if not treated properly. These complications are the third leading cause of death in Korea and are a serious problem.

In diabetes, drugs for the purpose of treatment have already been developed. Insulin preparations are effective for insulin-depent diabetes mellitus (IDDM), a type of autoimmune disease. Type 2 diabetes mellitus; non-insulin-dependent diabetes mellitus (NIDDM) currently uses sulphonylurease (tolbutamide, chloropropamide, glibenclamide, etc.) and biguanides (metformin). However, diabetes mellitus, along with the use of diet and rehydration therapy (rehydration therapy) is essential, in order to prevent the exacerbation of diabetes requires careful attention in daily life such as diet control and weight management. Therefore, it can be said that the development of a functional food having an effective effect in the treatment as well as the preventive effect by ingesting naturally as in the normal meal.

Regarding betulinic acid or betulinic acid derivatives, Korean Patent Publication No. 1020070093573 (Published: 2007.09.19, Registered Patent No. 1007636200000, Registered: 2007.09.27) 'Growth Hormone containing Sanjoin Extract or Betulinic Acid as an active ingredient The composition for promoting secretion 'describes the use of promoting the secretion of growth hormone of betulinic acid,

Republic of Korea Patent Application Publication No. 1020020092908 (Published: 2002.12.12, Registered Patent No. 1006181000000, Registered: 2006.08.23) 'New betulinic acid derivatives having anti-angiogenic activity, methods of preparing such derivatives and tumor-associated angiogenesis Their use for treatment 'describes the use of betulinic acid derivatives for the treatment of tumor-associated angiogenesis,

Republic of Korea Patent Publication No. 1020010080507 (published: 2001.08.22, Patent Application No. 10-2001-7006315, Filed: 2001.05.18) 'Betulinic acid derivatives, methods for their preparation and their use as cancer growth inhibitors' Although the use of lean acid derivatives for inhibiting cancer growth has been described, there is no suggestion or suggestion regarding the antidiabetic activity of betulinic acid or its derivatives.

The present inventors have found that betulinic acid and its derivatives have an excellent effect on alleviating the symptoms of diabetes, and concluded that betulinic acid and its derivatives can be used for the prevention and treatment of diabetes.

1. Republic of Korea Patent Publication No. 1020070093573 2. Republic of Korea Patent Publication No. 1020020092908 3. Republic of Korea Patent Publication No. 1020010080507

It is an object of the present invention to provide antidiabetic efficacy and utilization of betulin acid and its derivatives, and the present invention can be widely used in general and health foods, and also can be administered as a pharmaceutical composition through oral administration. To provide raw materials such as therapeutic and health supplements.

The present invention relates to the antidiabetic activity of betulinic acid and its derivatives.

Betulinic acid (IUPAC name: (3β) -3-Hydroxy-lup-20 (29) -en-28-oic acid) has the following formula (1).

In addition, the betulinic acid derivative according to the present invention has the following general formula (2) or (3):

The compound of Formula 2 has a C ₃₃ H ₄₈ N ₂ O ₂ formula and has a molecular weight of 504.75, and the compound of Formula 3 has a C ₃₃ H ₄₆ N ₂ O ₂ formula and has a molecular weight of 502.73.

According to the present invention, the compounds of Formulas 1 to 3 are characterized by having antidiabetic activity.

In addition, according to the present invention, there is provided an antidiabetic pharmaceutical composition or an antidiabetic functional health food composition comprising the compound of Formulas 1 to 3 as an active ingredient.

In the present invention, the term "active ingredient" refers to a substance or a group of substances (a pharmacologically active ingredient or the like, which is expected to express directly or indirectly the efficacy and effect of the drug by intrinsic pharmacological action. It means containing a main component).

The functional food composition may be provided in a conventional form (cookies, beverages, etc.) that will be apparent to those skilled in the art, including, but not limited to, betulinic acid, and acceptable admixtures in food preparation.

"Functional health food" as defined herein means food manufactured and processed using raw materials or ingredients with functional properties useful to the human body according to Act No. 6727 of the Health Functional Food Act. In addition, the term "functionality" means ingestion for the purpose of gaining useful fruit for health use such as nutrient control or physiological action on the structure and function of the human body.

In addition, the pharmaceutical composition may contain betulinic acid and other carriers or excipients, and may be provided in a form obvious to those skilled in the art such as tablets, pills, granules, liquids, capsules, and the like.

According to the present invention, betulinic acid and its derivatives have antidiabetic activity by inhibiting glucose tolerance. Betulinic acid and its derivatives of the present invention can be used as a functional food or food additives as well as intraperitoneal administration as well as oral administration to induce antidiabetic activity, and especially formulated as an injectable composition as well as an injectable composition. Has the possible advantages.

FIG. 1A is a photograph showing Western blotting of AMPK (Adenosine Monophosphate-activated Protein Kinase) phosphorylation of betulinic acid, and FIG. 1B is a phosphorylation result of betulinic acid derivatives of Formulas 2 and 3 It is a photograph shown by Western blotting.
Figure 2 is a graph showing the initial glucose ratio over time after insulin injection after untreated, AICAR treated, betulinic acid treated in diabetic induced mice.
FIG. 3 shows PGC-1α in mouse myoblasts by NT (None Treated), mistletoe extract (KM110), mistletoe chloroform extract (CHE), betulinic acid (BA), AICAR, and rasveratol (RSV). It is a graph showing the expression.
Figure 4 is a graph showing the expression of GLUT4 in mouse myoblasts by NT, KME (Korean Mistletoe Extract), CHE, BA, RSV, AICAR.
5 shows NT, NT + insulin, NT + bortmannin, NT + insulin + bortmannin, CHE, CHE + insulin, CHE + bortmannin, CHE + insulin + bortmannin, betulinic acid, Glucose acquisition in mouse myoblasts by betulinic acid + insulin, betulinic acid + bortmannin, betulinic acid + insulin + bortmannin, AICAR, AICAR + insulin, AICAR + bortmannin, AICAR + insulin + bortmannin The graph shown.
6 is a photograph showing AS160 phosphorylation in C57BL / 6 mouse EDL muscle by betulinic acid.
Figure 7 is a graph showing increased insulin secretion in rat pancreatic cells by various doses of NT, betulinic acid, KM110, AICAR, RSV, CHE.

Hereinafter, the present invention will be described in detail by way of examples, but the following examples are intended to illustrate the present invention, and it will be apparent to those skilled in the art that the scope of the present invention is not limited thereto.

This Example is experimental Example 1, which experiments AMPK phosphorylation by treating betulin acid and its derivatives in mouse myoblasts, Experimental Example 2 for measuring the effect of betulin acid on insulin sensitivity in a diabetic mouse model Experimental Example 3 measuring the effect of various AMPK activators including acid on PGC-1α expression, Experimental Example 4 measuring the effect of betulin acid and Korean mistletoe extract on GLUT4 expression in mouse myoblasts, Experimental Example 5 demonstrating that tulinic acid acquires glucose by a PI3K independent pathway, Experimental Example 6 measuring the effect of betulinic acid on AS160 expression in mouse myoblasts, and the effect of betulinic acid on insulin secretion in rat pancreatic cells It consists of Experimental Example 7, which measures. Through the above experimental example to evaluate the anti-diabetic efficacy of the betulinic acid of the present invention.

Experimental Example  1: in muscle cells Betulin  Acids and their derivatives AMPK  Effect on phosphorylation

C2C12 cells (mouse myoblasts) were used to confirm the antidiabetic effect of betulinic acid and its derivatives by AMPK phosphorylation. Betulinic acid and derivatives of Formulas 2 and 3, respectively, were treated with muscle at a concentration of 100 ug / ml and had specificity for phosphorylated AMPK (p-AMPK) after 30 minutes, 1 hour, 3 hours, and 6 hours. Western blotting with antibody. As a result, it was confirmed that it has a strong activity as shown in Figure 1a, Figure 1b.

Experimental Example  2: in diabetic-induced mice Betulin  Effect of Acids on Insulin Sensitivity

To determine the effect of betulinic acid on insulin sensitivity, 70 mpk (meaning 'mg per kg') means that 70 mg of absolute dose per kg of body weight of the subject was administered to 70 mpk in a 20-g mouse. Was used in mice induced with diabetes with alloxan).

Diabetes-induced mice pretreated or untreated with betulinic acid or AICAR (5-amino-4-imidazolecarboxamidiboside) 1 hour prior to insulin treatment were administered insulin and blood glucose was measured and the results are shown in FIG. 2. It was. Betulinic acid at 200mpk concentration was significantly different from the hypoglycemic level of diabetic mice that induced only the negative control group, and showed a higher hypoglycemic effect than the other group treated with insulin alone. In addition, it showed a hypoglycemic effect similar to AICAR of 200mpk concentration used as a positive control. This indicates that betulinic acid has an effect of lowering blood sugar by increasing insulin sensitivity.

Experimental Example  3: AMPK  Activator PGC Effect on -1α Expression

C2C12 cells were used to investigate the effects of betulinic acid on PGC-1α expression associated with insulin sensitivity. In this experiment, the drug AICAR with anti-diabetic effect by activating AMPK, resveratrol rich in red wine and grape skin, mistletoe extract (KM110) and chloroform extract (CHE) extracted from chloroform together with betulinic acid The expression level of PGC-1α was examined. As shown in FIG. 3, betulinic acid showed a stronger expression effect of PGC-1α than previously known AICAR or resveratrol.

Experimental Example  4: In muscle cells Betulinic acid  Korean Mistletoe Extract GLUT4  Effect on expression

To be metabolized, glucose in the blood must be transported by GLUT4 into each tissue. Therefore, GLUT4's role in glycemic control is very important. As shown in FIG. 4, mistletoe extracts and betulinic acid showed relatively lower expression levels than AICAR in GLUT4 expression in C2C12 cells, but higher anti-diabetic effects than well-known resveratol.

Experimental Example  5: In muscle cells Betulin  Mountains PI3K Independent  Glucose Acquisition by Pathway uptake )

In order to control blood glucose, cells must acquire glucose and in response, insulin is secreted to bind to its receptors and then through various pathways. There are several pathways but they are usually caused by PI3K-dependent pathways, that is, PI3K pathways, but can also occur in PI3K-independent pathways. In this experiment, various samples including betulinic acid were treated to C2C12 cells to confirm 2-deoxyglucose and PI3K inhibitor, buttmannin, which were recognized as ³ H, and obtained the results as shown in FIG. 5. The results suggest that betulinic acid and CHE may be induced by PI3K-independent pathways such as AICAR. It has been demonstrated that glycemic control is achieved.

Experimental Example  6: AS160 For Betulinic  effect

AMPK activator drug AICAR with antidiabetic efficacy phosphorylates AS160 to obtain glucose. Accordingly, betulinic acid was activated to phosphorylate AS160 like AICAR in the presence of wortmannin to obtain a result as shown in FIG. 6. As shown, betulinic acid phosphorylated AS160, which showed antidiabetic effect by phosphorylating AS160 by PI3K independent pathway.

Experimental Example  7: Betulin  Effect of acid on insulin secretion

The effects of betulinic acid, mistletoe extract and other drugs on insulin secretion of RINm5F cells (rat pancreatic β-cells) were examined to obtain results as shown in FIG. 7. In particular, the Korean mistletoe extract CHE significantly increased insulin secretion, and betulinic acid and other drugs increased concentration-dependently. This result suggests that Korean mistletoe extract and betulinic acid promote insulin secretion like AICAR or resveratrol. This can be an example of the effect.

Claims (3)

Betulinic acid or a derivative thereof of Formulas 1 to 3 characterized by having antidiabetic activity:
Formula 1

Formula 2

(3)

A pharmaceutical composition for preventing or treating diabetes, comprising betulinic acid or a derivative thereof as an active ingredient. Functional health food composition for diabetic symptoms improvement comprising the betulinic acid or derivatives of claim 1 as an active ingredient.

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