Use of Antrodia camphorata compounds in the preparation of pharmaceutical compositions for the treatment of diabetes

Abstract

The purpose of this invention is to provide the use of an Antrodia camphorata compound in the preparation of pharmaceutical compositions for treating diabetes, wherein the Antrodia camphorata compound is as shown in the following chemical formula 1: Or, a pharmacologically acceptable salt of chemical formula 1.

Description

Use of Antrodia camphorata compounds in the preparation of a pharmaceutical composition for treating diabetes The present invention discloses a use of Antrodia camphorata compounds, in particular a use for preparing a pharmaceutical composition for treating diabetes. Antrodia camphorata, also known as Zhangzhi, Niuzhanggu, Hongzhangzhi, etc., belongs to the perennial mushroom of Aphyllophorales and Polyporaceae, and is an endemic fungus in Taiwan. It only grows on the inner wall of the hollow and decayed heartwood of Cinnamoum kanehirai Hay, a protected tree species in Taiwan. Due to the extremely scarce distribution of Cinnamoum kanehirai Hay and the illegal logging by humans, the number of wild Antrodia camphorata that can grow parasitically in it is even scarcer. Moreover, because its fruiting body grows quite slowly and the growth period is only between June and October, the price is very expensive. Among the many components of Antrodia cinnamomea, triterpenoid compounds have been studied the most. Triterpenoid compounds are a general term for natural compounds composed of thirty carbon elements combined into hexagonal or pentagonal shapes. The bitterness of Antrodia cinnamomea mainly comes from this component of triterpenoids. In 1995, Cherng et al. found that the fruiting body extract of Antrodia cinnamomea contained three new triterpenoid compounds with an ergostane skeleton: antcin A, antcin B, and antcin C (Cherng, I. H., and Chiang, H. C. 1995. Three new triterpenoids from Antrodia cinnamomea. J. Nat. Prod. 58:365-371). Chen et al. found three triterpenoid compounds, namely zhankuic acid A, zhankuic acid B, and zhankuic acid C, after extracting the fruiting body of Antrodia cinnamomea with ethanol (Chen, C. H., and Yang, S. W. 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 found three other new triterpenoid compounds from the fruiting body extract in 1995, namely a sesquiterpene lactone and two bisphenol derivatives, namely antrocin, 4,7-dimethoxy-5-methyl-1,3-benzodioxole, and 2,2',5,5'-tetramethoxy-3,4,3',4'-bi-methylenedioxy-6,6'-dimethylbiphenyl (Chiang, H. C., Wu, D. P., Cherng, I. W., and Ueng, C. H. 1995. A sesquiterpene lactone, phenyl and biphenyl compounds from Antrodia cinnamomea. Phytochemistry. 39:613-616).By 1996, Cherng et al. again discovered four new triterpenoid compounds, namely antcin E, antcin F, methyl antcinate G, and methyl antcinate H, using the same analytical method (Cherng, I. H., Wu, D. P., and Chiang, H. C. 1996. Triteroenoids from Antrodia cinnamomea. Phytochemistry. 41:263-267). Yang et al. discovered two new compounds with ergostane skeletons, namely zhankuic acid D and zhankuic acid E, and three new compounds with lanostane skeletons: 15α-acetyl-dehydrosulphurenic acid, dehydroeburicoic acid, and dehydrasulphurenic acid (Yang, S. W., Shen, Y. C., and Chen, C. H. 1996. Steroids and triterpenoids of Antrodia cinnamomea-a fungus parasitic on Cinnamomum micranthum. Phytochemistry. 41:1389-1392). Diabetes belongs to chronic endocrine and metabolic diseases. It is a disease caused by absolute or relative insufficiency of insulin secretion, resulting in disorders of sugar, fat, and protein metabolism. Clinically, it is divided into two types: type I and type II. Type I is insulin-dependent (IDDM), and the vast majority of cases are due to autoimmunity. Patients' pancreatic β-cells are damaged and unable to secrete insulin, resulting in absolute insulin deficiency. Type II is non-insulin-dependent (NIDDM). Patients' pancreatic β-cells still have the function of secreting insulin, but due to the resistance or insensitivity of insulin receptors on target cells (corresponding cells) 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 need to maintain normal metabolism, leading to elevated blood sugar and increased urine sugar. This situation is called relative insulin insufficiency. Taking the invention patents previously applied by the applicant as examples, such as the invention patent of the Republic of China Patent Publication No. I527583, "Pharmaceutical Composition for Promoting Wound Healing and Its Use", the invention patent of the Republic of China Patent Publication No. I626053, "Use of Antrodia camphorata Compounds for Preparing a Composition for Promoting Hair Growth", the invention patent of the People's Republic of China Patent Publication No. CN104887661B, "Pharmaceutical Composition for Promoting Wound Healing and Its Use", and the patent application number of the People's Republic of China CN201610313277.9, "Use of Antrodia camphorata Extract and Compounds for Preparing a Composition for Promoting Hair Growth" as examples, they have disclosed the compound 4,7-dimethoxy-5-methyl-1,3-benzodioxole (hereinafter referred to as SY1) purified from Antrodia camphorata, which has the effects of promoting wound healing and promoting hair growth, as well as the potential to develop related pharmaceutical compositions. However, whether this compound has other medical applications remains to be studied. So far, no literature has disclosed that this compound can be applied to related uses such as reducing blood sugar and blood lipids to improve type 2 diabetes. Accordingly, based on extensive research on Antrodia camphorata compounds, after painstaking research and repeated experiments, the inventors gradually found relevant applications in specific aspects, such as the related uses for improving type 2 diabetes by reducing blood sugar, blood lipids, etc. disclosed in this case. The object of the present invention is to provide a use of an Antrodia camphorata compound for preparing a pharmaceutical composition for treating diabetes, wherein the Antrodia camphorata compound is as shown in the following Chemical Formula 1: Chemical Formula 1, or a pharmaceutically acceptable salt of the Chemical Formula 1. Preferably, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier. Preferably, the diabetes is type 2 diabetes. Preferably, it further comprises reducing the blood sugar of a patient suffering from the diabetes. Preferably, it further comprises reducing the blood lipids of a patient suffering from the diabetes. Antrodia camphorata extract SY-C and the compound SY-1 for the preparation Take the mycelium, fruiting body of Antrodia camphorata, or a mixture of both, and extract with water or an organic solvent to obtain an aqueous extract or an organic solvent extract of Antrodia camphorata. First, take 1.0 kg of the mycelium, fruiting body of Antrodia camphorata, or a mixture of both, extract it twice with 10 times the amount of alcohol, and then combine and concentrate it to obtain approximately 230 g of a crude extract. Further subject the aforementioned crude extract to three times of partition extraction with a dichloromethane / water (1:1) solution to divide it into a dichloromethane layer of approximately 102.6 g and a water layer of approximately 127.4 g. Take 6.0 g of the dichloromethane layer and separate it by silica gel column chromatography with solvents of n-hexane / dichloromethane (1:4), dichloromethane, and methanol / dichloromethane (5:95) to obtain four fractions, namely LT-E-D-1, LT-E-D-2, LT-E-D-3, and LT-E-D-4. The acquisition of the above four fractions is as follows: After eluting with n-hexane / dichloromethane (1:4) and dichloromethane eluents respectively and combining the obtained fractions, it is LT-E-D-1. Then elute with a methanol / dichloromethane (5:95) eluent. The fraction obtained in the first half is LT-E-D-2, and the fraction obtained in the second half is LT-E-D-3. Finally, elute with methanol to obtain LT-E-D-4. Among them, the LT-E-D-1 fraction, after recovery, is the Antrodia camphorata extract SY-C. To further purify the active ingredients in the Antrodia camphorata extract SY-C, take out the LT-E-D-1 layer, and then it can be separated and purified by silica gel column chromatography or by a preparative high performance liquid chromatograph to purify the compound 4,7-dimethoxy-5-methyl-1,3-benzodioxole shown in Chemical Formula 1 (i.e., SY-1). Chemical Formula 1 Among them, the organic solvent can include alcohols (such as methanol, ethanol, or propanol), esters (such as ethyl acetate), alkanes (such as hexane), or haloalkanes (such as chloromethane, chloroethane), but is not limited thereto. Among them, the preferred one is alcohols, and the more preferred one is ethanol. The weight ratio of the organic solvent to the mycelium, fruiting body, or a mixture of both can be 15:1 to 5:1, preferably 12:1 to 8:1, but is not limited thereto. In addition, the ratio of the dichloromethane / water solution can be between 2:1 and 1:2, and is not limited thereto. Regarding the extraction of 4,7-dimethoxy-5-methyl-1,3-benzodioxole, reference can be made to that described in Tu SH., J Agric FoodChem. 2012 Apr 11;60(14):3612-8. In previous related studies, whether it was the extract of Antrodia cinnamomea or the purified SY-1 compound, the focus was mainly on its anti-tumor effect, and there was no research on the treatment of diabetes. In this invention, experiments were conducted using the compound SY-1 purified from Antrodia cinnamomea and its salts, and it was found and confirmed that both have the effect of promoting insulin secretion, which belongs to a novel use that has never been seen before. Animal model of type 2 diabetic mice In this experiment, four-week-old male C57BL / 6JNarl mice from the National Animal Experiment Center (Taipei, Taiwan) were used and bred in an environment of 23°C ± 2°C with a 12-hour light / 12-hour dark cycle. The mice were allowed to adapt for one week and were fed water and food ad libitum. After one week, they were divided into groups and fed a commercial feed 1320 (containing 11% fat) purchased from Jieyi Enterprise Co., Ltd. (New Taipei City, Taiwan) and D12451 (containing 45% high-fat feed) purchased from Meijing Biotechnology Co., Ltd. (Taichung, Taiwan) for 12 weeks to induce diabetes. The water and feed intake were calculated weekly, and the bedding urine volume was observed to monitor whether there were any changes in the status of the mice (data not shown), so as to know whether the mice had reached the diabetic model. During the experiment, the body weight and fasting blood glucose value of the mice were measured weekly and the changes were recorded. The mean ± 3SD of the fasting blood glucose value of the mice was calculated. If the fasting blood glucose value exceeded the mean + 3SD of the normal diet group, intragastric administration of the drug was started for 4 weeks, and a glucose tolerance test was performed by collecting blood from the tail vein in the last week of the experiment. On the last day of the experiment, all animals (except for those that died unexpectedly) were fasted for 20 hours at the National Animal Experiment Center, euthanized with pure carbon dioxide, and autopsy was performed, including weighing the organs (liver) and a rough examination, including the evaluation of the liver and pancreas. Most of the lesions of the animals were recorded at the time of final sacrifice, including the scheduled euthanasia and unscheduled deaths. Experimental design and methods Table 1: Experimental groups and drug administration treatment conditions Glucose tolerance test ( The last week ) Please refer to Figures 1 - 2, which respectively show the test results of the Oral Glucose Tolerance Test (OGTT). Each group shown in Figures 1 - 2 corresponds 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). Among them, NFD refers to a normal fat diet, HFD refers to a high - fat diet, and Glibenclamide is an oral hypoglycemic drug for type 2 (non - insulin - dependent) diabetic patients. The results show that according to the results of the Oral Glucose Tolerance Test (OGTT) at the 16th week, it can be seen that for the group with the addition of the hypoglycemic drug Glibenclamide, when no sugar water was fed, the fasting blood glucose increased higher than any other group. Thus, it can be known that after 4 weeks of drug treatment, the hypoglycemic effect of this drug is worse in the later stage compared to SY1. Regarding the part of SY1 feeding, 30 minutes after feeding sugar water, SY1 had an obvious hypoglycemic effect in the low - dose and medium - dose groups of HFD + 5 and HFD + 10. In the HFD + 5 group, the hypoglycemic effect was comparable to that of the HFD + 10 group (n = 6, the average blood glucose at 120 minutes in HFD + 5 and HFD + 10 was 155.83 and 155.33 respectively), and the effect was more obvious than that of the high - dose HFD + 20 and the hypoglycemic drug Glibenclamide groups. in the blood ALT and AST biochemical analysis Two transaminases in the blood, including Alanine aminotransferase (ALT) and Aspartate aminotransferase (AST). ALT is mainly present in liver cells, while AST is mainly present in the liver, myocardium, muscles, and also in red blood cells. When these cells are damaged and necrotic for various reasons, the intracellular ALT and AST will be released into the blood. Therefore, in cases of hepatitis, myocardial infarction, muscle inflammation, or hemolysis, the AST value may increase. By抽血检验 (drawing blood for testing) the degree of increase of these enzymes, the degree of cell damage can be inferred; similarly, an increase in the ALT value can be said to be caused by liver inflammation. The liver function can be evaluated by detecting these two enzymes. In this experiment, a Hitachi 7080 blood biochemical analyzer and related reagents were used, and the testing was entrusted to the National Laboratory Animal Center. The results are shown in Figures 3 to 4, which respectively show the biochemical analysis results 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 among 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, while this phenomenon was not seen in the SY1 group during the same treatment period. Low-density lipoprotein ( Low-density lipoprotein, LDL ) detection Low-density lipoprotein (LDL) is one of the five main groups of lipoproteins. LDL is a large spherical macromolecule that can transport hydrophobic molecules (lipids and cholesterol) throughout the body through the aqueous environment of the blood. Low-density lipoprotein LDL delivers fat molecules to cells, and if they are oxidized within the arterial wall, it may cause atherosclerosis. Since oxidized low-density lipoproteins are more easily retained by proteoglycans, there is 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. In the part of the LDL test results, the effect of reducing LDL in the HFD+5 group was not much different from that of the Western medicine group and was also better than that of the HFD+10 group. This repeatedly proves that the HFD+5 group is the best dosage for both blood sugar reduction and LDL reduction. The use of the Ganoderma lucidum compound provided by the present invention for preparing a pharmaceutical composition for treating diabetes indeed has industrial utilization value. However, the above description is only for the illustration of the preferred embodiments of the present invention. Those skilled in this art can make various other improvements based on the above description, but these changes still fall within the spirit of the present invention and the patent scope defined below. None Figures 1 to 2 respectively show the results of the oral glucose tolerance test (OGTT); Figure 3 shows the results of the biochemical test of glutamate phenylacetate transaminase (AST); Figure 4 shows the results of the biochemical test of alanine transaminase (ALT); Figure 5 shows the results of the biochemical test of low-density lipoprotein (LDL). None

Claims

1. Use of an Antrodia camphorata compound in the preparation of a pharmaceutical composition for the treatment of diabetes, wherein the Antrodia camphorata compound is as shown in Chemical Formula 1 below: Chemical Formula 1, or a pharmacologically acceptable salt of Chemical Formula 1.

2. The use as described in claim 1, wherein the pharmaceutical composition further includes a pharmaceutically acceptable carrier.

3. The use as described in claim 1, wherein the diabetes is type 2 diabetes.

4. The uses described in claim 3 further include lowering blood sugar levels in patients with the diabetes.

5. The uses described in claim 3 further include lowering blood lipids in patients with the diabetes.

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