Application of antrodia camphorata compound in preparation of pharmaceutical composition for treating diabetes

Abstract

The invention provides application of an antrodia camphorata compound to preparation of a pharmaceutical composition for treating diabetes mellitus. The antrodia camphorata compound is shown in the following chemical formula 1: # imgabs0 # chemical formula 1, or pharmacologically acceptable salts of the chemical formula 1.

Description

Technical Field

[0001] The present invention discloses a use of an Antrodia cinnamomea compound, and particularly relates to a use for preparing a pharmaceutical composition for treating diabetes. Background Art

[0002] Antrodia camphorata, also known as Antrodia camphorata, Antrodia camphorata mushroom or red Antrodia camphorata, belongs to the perennial fungus of the Aphyllophorales and Polyporaceae families. It is endemic to Taiwan Province, China and only grows on the inner wall of the hollow rotten heartwood of the protected tree species Cinnamoum kanehirai Hay in Taiwan Province, China. Due to the extremely rare distribution of Cinnamoum kanehirai Hay and the illegal logging, the number of wild Antrodia camphorata that can only grow in it is even rarer. In addition, because its fruiting body grows very slowly and the growing period is only between June and October, the price is very expensive.

[0003] Among the many components of Antrodia cinnamomea, triterpenoids are the most studied. Triterpenoids are a general term for natural compounds composed of thirty carbon elements combined into hexagonal or pentagonal shapes. The bitter taste of Antrodia cinnamomea mainly comes from triterpenoids. In 1995, Cherng et al. found that the Antrodia cinnamomea fruiting body extract contained three new triterpenoids with ergostane as the skeleton: antcin A, antcin B and antcin C (Cherng, IH, and Chiang, HC 1995. Three new triterpenoids from Antrodiacinnamomea. J. Nat. Prod. 58: 365-371). Chen et al. extracted the fruiting bodies of Antrodia cinnamomea with ethanol and found three triterpenoid compounds, namely, zhangkuic acid A, zhangkuic acid B and zhangkuic acid C (Chen, CH, and Yang, SW 1995. New steroid acids from Antrodia cinnamomea, - a fungus parasitic on Cinnamomum micranthum. J. Nat. Prod. 58: 1655-1661). In addition, Chiang et al. also discovered three other new triterpenoids from the fruiting body extracts, namely, antrocin, 4,7-dimethoxy-5-methyl-1,3-benzodioxole and 2,2',5,5'-teramethoxy-3,4,3',4'-bi-methylenedioxy-6,6'-dimethylbiphenyl (Chiang, HC, Wu, DP, Cherng, IW, and Ueng, CH1995. Asesquiterpene lactone, phenyl and biphenyl compounds from Antrodiacinnamomea. Phytochemistry. 39:613-616).In 1996, Cherng et al. discovered four new triterpenoids using the same analytical method: antcin E, antcin F, methyl antcinate G, and methylantcinate H (Cherng, IH, Wu, DP, and Chiang, HC 1996. Triteroenoids from Antrodia cinnamomea. Phytochemistry. 41: 263-267); while Yang et al. discovered two new compounds with ergostane as the skeleton, zhankuic acid D and zhankuic acid E, and three new compounds with lanostane as the skeleton: 15α-acetyl-dehydrosulphurenic acid, dehydroeburicoic acid, and dehydrasulphurenic acid (Yang, SW, Shen, YC, and Chen, CH 1996. Steroids and triterpenoids ofAntrodia cinnamomea—a fungus parasitic on Cinnamomummicranthum. Phytochemistry. 41:1389-1392).

[0004] Diabetes is a chronic endocrine metabolic disease. It is a disease caused by absolute or relative deficiency of insulin secretion, which leads to disorders in sugar, fat and protein metabolism. It is clinically divided into type I and type II. Type I is insulin-dependent (IDDM), which is mostly caused by autoimmunity. The patient cannot secrete insulin due to damage to the pancreatic β cells, resulting in absolute insulin deficiency; type II is non-insulin-dependent (NIDDM). The patient's pancreatic β cells still have the function of secreting insulin, but because the target cell (corresponding cell) insulin receptors are resistant or insensitive to insulin, insulin is not released when needed or the binding force between the receptor and insulin decreases. Although the amount of insulin is normal, it cannot meet the needs of maintaining normal metabolism, resulting in increased blood sugar and increased urine sugar. This situation is called relative insulin deficiency.

[0005] Taking the invention patents previously applied by the applicant as examples, such as the invention patent of Taiwan Province of China Patent Publication No. I527583 "Medical composition for promoting wound healing and its use", the invention patent of Taiwan Province of China Patent Publication No. I626053 "Use of Antrodia cinnamomea compound for preparing a composition for promoting hair growth", the invention patent of China Patent Publication No. CN104887661B "Medical composition for promoting wound healing and its use" and the Chinese patent application No. CN201610313277.9 "Use of Antrodia cinnamomea extract and compound for preparing a composition for promoting hair growth", they have disclosed the compound 4,7-dimethoxy-5-methyl-1,3-benzodioxole (SY1) purified from Antrodia cinnamomea, which has the effects of promoting wound healing and promoting hair growth, and the potential for developing related pharmaceutical compositions. However, whether the compound has other medical applications remains to be studied. So far, there is no literature revealing that the compound can be used to lower blood sugar, blood lipids, and improve type 2 diabetes. Summary of the invention

[0006] Based on this, the inventors have made considerable research on Antrodia cinnamomea compounds, and after painstaking research and repeated experiments, they have gradually found relevant applications in specific aspects, such as the use disclosed in this case that can reduce blood sugar, blood lipids, etc. to improve type 2 diabetes.

[0007] The object of the present invention is to provide a use of an Antrodia cinnamomea compound for preparing a pharmaceutical composition for treating diabetes, wherein the Antrodia cinnamomea compound is as shown in the following chemical formula 1:

[0008] Chemical formula 1, or a pharmacologically acceptable salt of Chemical formula 1.

[0009] Preferably, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier.

[0010] Preferably, the diabetes is type 2 diabetes.

[0011] Preferably, the method is used to lower blood sugar in patients suffering from diabetes.

[0012] Preferably, the method is further used for lowering blood lipids in patients suffering from diabetes. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] Figure 1 Displays the results of the glucose tolerance test (OGTT);

[0014] Figure 2 Displays the results of the glucose tolerance test (OGTT);

[0015] Figure 3 Displays the results of biochemical test of glutamate phenylacetate transaminase (AST);

[0016] Figure 4 Displays the results of alanine transaminase (ALT) biochemical test;

[0017] Figure 5 Displays the results of low-density lipoprotein (LDL) biochemical tests. DETAILED DESCRIPTION

[0018] Preparation of Antrodia cinnamomea extract SY-C and compound SY-1

[0019] Take the mycelium, fruiting body or a mixture of Antrodia camphorata and extract them with water or an organic solvent to obtain Antrodia camphorata water extract or organic solvent extract. First, take 1.0 kg of Antrodia camphorata mycelium, fruiting body or a mixture of the two and extract them twice with 10 times the amount of alcohol, then combine and concentrate them to obtain a crude extract of about 230 g. The aforementioned crude extract is further subjected to a three-time distribution extraction method with a dichloromethane / water (1:1) solution to separate it into a dichloromethane layer of about 102.6 g and a water layer of about 127.4 g. Take 6.0 g of the dichloromethane layer and separate it with a silica gel column chromatography method through a solvent separation of n-hexane / dichloromethane (1:4), dichloromethane, and methanol / dichloromethane (5:95), and it can be separated into four fractions (fractions) such as LT-ED-1, LT-ED-2, LT-ED-3, and LT-ED-4. The above four fractions are obtained as follows: after being extracted with n-hexane / dichloromethane (1:4) and dichloromethane respectively, the fractions obtained are combined to obtain LT-ED-1, and then extracted with methanol / dichloromethane (5:95) to obtain the first half of the fraction as LT-ED-2, the second half of the fraction as LT-ED-3, and finally extracted with methanol to obtain LT-ED-4. Among them, the LT-ED-1 fraction is recovered as Antrodia cinnamomea extract SY-C.

[0020] In order to further purify the active ingredients of Antrodia cinnamomea extract SY-C, the LT-ED-1 layer is taken out and then separated and purified by silica gel column chromatography or preparative high performance liquid chromatography to purify the compound 4,7-dimethoxy-5-methyl-1,3-benzodioxole represented by the following chemical formula 1 (i.e., SY-1).

[0021]

[0022] The organic solvent may include alcohols (such as methanol, ethanol or propanol), esters (such as ethyl acetate), alkanes (such as hexane) or halogenated alkanes (such as methyl chloride, ethyl chloride), but is not limited thereto. Alcohols are preferred, and ethanol is more preferred. The weight ratio of the organic solvent to the mycelium, the fruiting body or the mixture of the two may be 15:1 to 5:1, preferably 12:1 to 8:1, but is not limited thereto. In addition, the ratio of dichloromethane to aqueous solution may be between 2:1 and 1:2, but is also not limited thereto.

[0023] For the extraction of 4,7-dimethoxy-5-methyl-1,3-benzodioxole, reference may be made to Tu SH., J Agric Food Chem. 2012 Apr 11; 60(14): 3612-8.

[0024] In previous related studies, whether it is Antrodia cinnamomea extract or purified SY-1 compound, the main focus is on its anti-tumor effect, and there is no research on the treatment of diabetes. The present invention uses the purified Antrodia cinnamomea compound SY-1 and its salts to conduct experiments and finds and confirms that they both have the effect of promoting insulin secretion, which is a novel use that has never been seen before.

[0025] Animal model of type 2 diabetes mellitus in mice

[0026] Four-week-old male C57BL / 6JNarl mice were obtained from the Animal Experiment Center (Taipei, Taiwan Province, China) and were housed at 23°C ± 2°C with a 12-h light / 12-h dark cycle.

[0027] The mice were fed water and food ad libitum for one week after adaptation. One week later, they were divided into groups and fed with commercial diet 1320 (containing 11% fat) purchased from Jie Yi Enterprise Co., Ltd. (New Taipei City, Taiwan Province, China) and D12451 (containing 45% high-fat diet) purchased from Meijing Biological Co., Ltd. (Taichung, Taiwan Province, China) for 12 weeks of induction.

[0028] The amount of water and feed was calculated weekly and the amount of urine in the litter was observed to monitor whether the mice's condition changed (data not shown) to determine whether the mice had reached the diabetic model.

[0029] During the experiment, the body weight and fasting blood glucose level of mice were measured weekly and their changes were recorded. The mean±3SD of the fasting blood glucose level of mice was calculated. If the fasting blood glucose level exceeded the mean+3SD of the normal diet group, tube feeding was started for 4 weeks. In the last week of the experiment, blood was collected from the tail vein for glucose tolerance test.

[0030] On the last day of the experiment, all animals (except unplanned deaths) were fasted for 20 h and euthanized with pure carbon dioxide at the Animal Experiment Center, and necropsy, organ weight (liver) and gross examination, including evaluation of the liver and pancreas, were performed. Most lesions of the animals were recorded at the time of final sacrifice, including scheduled euthanasia and unscheduled deaths.

[0031] Experimental design and methods

[0032] Table 1: Experimental groups and treatment conditions

[0033]

[0034]

[0035] Glucose tolerance test (final week)

[0036] See also Figures 1-2 , Figures 1-2 The test results of the Oral Glucose Tolerance Test (OGTT) are shown respectively. Figures 1-2 The groups shown correspond to the conditions shown in Table 1: Group A (NFD), Group B (NFD+5), Group C (NFD+20), Group D (HFD), Group E (HFD+5), Group F (HFD+10), Group G (HFD+20), Group H (Glibenclamide). NFD refers to a normal fat diet, HFD refers to a high fat diet, and Glibenclamide is an oral hypoglycemic drug for patients with type II (non-insulin-dependent) diabetes.

[0037] The results showed that according to the results of the oral glucose tolerance test (OGTT) at week 16, the group that added the hypoglycemic drug Glibenclamide had a higher increase in fasting blood sugar than any other group when not fed with sugar water. This shows that after 4 weeks of drug treatment, the blood sugar lowering effect of this drug was poorer than that of SY1 in the later stage of treatment.

[0038] In the SY1 feeding part, 30 minutes after feeding sugar water, SY1 had a significant effect on lowering blood sugar in the low-dose and medium-dose groups of HFD+5 and HFD+10. The blood sugar lowering effect in the HFD+5 group was comparable to that in the HFD+10 group (n=6, the average 120-minute blood sugar levels of HFD+5 and HFD+10 were 155.83 and 155.33, respectively), and the effect was more significant than that of the high-dose HFD+20 and hypoglycemic drug Glibenclamide groups.

[0039] Biochemical analysis of ALT and AST in blood

[0040] There are two types of transaminases in the blood, including alanine aminotransferase ALT (Alanine aminotransferase) and aspartate aminotransferase AST (aspartate aminotransferase). ALT is mainly found in liver cells, while AST is mainly found in the liver, myocardium, muscles, and red blood cells. When these cells are necrotic and damaged due to various reasons, the ALT and AST in the cells will be released into the blood. Therefore, when there is hepatitis, myocardial infarction, muscle inflammation or hemolysis, the AST value may increase. The degree of increase in these enzymes can be used to infer the degree of cell damage by drawing blood to test the increase in these enzymes. Similarly, an increase in ALT can be said to be caused by liver inflammation. Liver function can be evaluated by testing the above two enzymes.

[0041] In this experiment, Hitachi 7080 blood biochemical analyzer and related reagents were used and tested by the Experimental Animal Center. Figures 3-4 As shown in the figure, it shows the results of biochemical analysis of aspartate transaminase (AST) and alanine transaminase (ALT) in the blood. In terms of liver function in the late stage of treatment, there was little difference between the groups, but the liver function index of the western medicine group increased and reached a significant difference. Long-term use of this western medicine may cause slight damage to the liver, but this phenomenon was not seen in the SY1 group during the same treatment period.

[0042] Low-density lipoprotein (LDL) test

[0043] Low-density lipoprotein (LDL) is one of the five major groups of lipoproteins. LDL is a large spherical molecule that transports hydrophobic molecules (lipids and cholesterol) throughout the body through the aqueous environment of the blood. LDL delivers fat molecules to cells, and if they are oxidized within the artery wall, they may produce atherosclerosis. Because oxidized LDL is more easily retained by proteoglycans, they pose a risk of cardiovascular disease when they invade the endothelium. Therefore, high levels of LDL are associated with arterial inflammation, leading to cardiovascular diseases (CVD) such as atherosclerosis, ischemic stroke, and myocardial infarction.

[0044] In terms of LDL test results, the HFD+5 group had an LDL-lowering effect that was not much different from that of the Western medicine group, and was better than HFD+10. This has repeatedly proved that the HFD+5 group is the optimal dose for both lowering blood sugar and LDL.

[0045] The use of the Antrodia cinnamomea compound provided by the present invention for preparing a pharmaceutical composition for treating diabetes is indeed of industrial value. However, the above description is only an illustration of a preferred embodiment of the present invention. Anyone skilled in the art can make other various improvements based on the above description, but these changes still fall within the spirit of the present invention and the scope of the patent defined below.

[0046]

Explanation of symbols

[0047] none

[0048]

Biological material storage

[0049] none

Claims

1. A use of an Antrodia cinnamomea compound for preparing a pharmaceutical composition for treating diabetes, characterized in that: The Antrodia cinnamomea compound is shown in the following chemical formula 1: Or a pharmacologically acceptable salt of the chemical formula 1.

2. The use according to claim 1, characterized in that The pharmaceutical composition comprises a pharmaceutically acceptable carrier.

3. The use according to claim 1, characterized in that The diabetes mellitus is type 2 diabetes mellitus.

4. The use according to claim 3, characterized in that This includes use in lowering blood sugar in patients suffering from such diabetes.

5. The use according to claim 3, characterized in that: Including use for lowering blood lipids in patients suffering from such diabetes.

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