WO2016145982A1 - 鼠尾草酸的医药用途 - Google Patents

鼠尾草酸的医药用途 Download PDF

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WO2016145982A1
WO2016145982A1 PCT/CN2016/074780 CN2016074780W WO2016145982A1 WO 2016145982 A1 WO2016145982 A1 WO 2016145982A1 CN 2016074780 W CN2016074780 W CN 2016074780W WO 2016145982 A1 WO2016145982 A1 WO 2016145982A1
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petroleum ether
rosemary
composition
extract
group
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French (fr)
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李萍
徐晓军
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China Pharmaceutical University
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China Pharmaceutical University
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Priority claimed from CN201510120789.9A external-priority patent/CN106138024A/zh
Priority claimed from CN201510120866.0A external-priority patent/CN106138025B/zh
Priority claimed from CN201510540350.1A external-priority patent/CN105878366A/zh
Priority claimed from CN201510942960.4A external-priority patent/CN106309546A/zh
Application filed by China Pharmaceutical University filed Critical China Pharmaceutical University
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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
    • A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
    • A61K31/19—Carboxylic acids, e.g. valproic acid
    • A61K31/192—Carboxylic acids, e.g. valproic acid having aromatic groups, e.g. sulindac, 2-aryl-propionic acids, ethacrynic acid 

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  • the invention discloses a field of natural medicine, discloses a medical use of carnosic acid, and discloses the use of carnosic acid for preventing or treating type I diabetes and/or diabetic nephropathy, and the prevention or treatment of rosemary petroleum ether extract is high. Uses for lipemia, hyperglycemia, nonalcoholic fatty liver disease, diabetic nephropathy, and/or type 2 diabetes.
  • Diabetes is a metabolic disease characterized by hyperglycemia caused by defects in insulin secretion and/or its biological dysfunction. It is a genetic susceptibility disease with environmental factors involved in genetics. It is a chronic systemic disease. Metabolic diseases, which seriously endanger human health, and actively carry out prevention and treatment of diabetes have become major social public health problems.
  • Type II diabetes is a pathological process from insulin resistance with insulin deficiency to insulin secretion deficiency with insulin resistance.
  • Type II diabetes is currently recognized as a complex genetic disease caused by multiple factors or interactions caused by environmental factors and lifestyle changes, but the etiology of type 2 diabetes has not yet been fully elucidated.
  • Western medicine currently uses diet, exercise therapy sulfonylureas, biguanides, thiazolidinediones, ⁇ -glucosidase inhibitors, repaglinide, insulin, etc. to treat type 2 diabetes, only thiazolidinediones Improve insulin resistance. While the patients are treated with western medicine, complications such as arteriosclerosis, coronary heart disease and hypertension are constantly appearing. The complex mechanism of type II diabetes and the systemic diseases caused by it are the weak links of western medicine treatment. Whether it is a sulphur-promoting agent or a thiazolidinedione sensitizer, it has no obvious weight loss effect in experimental and clinical research, and obesity is an important pathological basis of type 2 diabetes and even metabolic syndrome.
  • Type II diabetes is not only a disorder of glucose metabolism, but also a disorder of fat metabolism.
  • People with type 2 diabetes caused by obesity usually have a high-sugar and high-fat diet and are often accompanied by hyperlipidemia.
  • most obese people do not develop diabetes, indicating that the body's own condition plays an important role. Under the same toxic effects, susceptible people can easily develop into type II diabetes, and insensitive people may not develop disease for a lifetime or delay the onset of disease.
  • Modern medicine can't change the susceptibility of the body, but it can reduce the early incentives as much as possible by changing lifestyles, losing weight, etc., and prolonging the compensation period as much as possible, thus delaying the occurrence of type 2 diabetes and even avoiding the onset.
  • the classification standard formulated by the Ministry of Health formulated the "Guidelines for the Clinical Research of New Drugs for Treating Diabetes Mellitus (Diabetes)" is the most widely used classification method, namely, Yin Deficiency Syndrome, Qi and Yin Deficiency Syndrome, Yin and Yang. Two types of deficiency syndrome and blood stasis syndrome.
  • Diabetic nephropathy is one of the most important complications of diabetes.
  • the incidence rate in China is also on the rise. It has become the second leading end stage renal disease, second only to glomerulonephritis, due to the complexity.
  • Metabolic disorders, kidney damage in diabetes, especially once entering the clinical proteinuria the condition is generally irreversible, often progressive development until end-stage renal disease, often more difficult than other kidney diseases, is the main cause of death in diabetic patients One of them, the pathogenesis of DN and the development of therapeutic drugs have received great attention from the medical community.
  • Modern medicine begins with diet control, blood sugar control, blood pressure reduction, lipid metabolism adjustment, or the use of dialysis, kidney transplantation and other treatments, there is no effective Western medicine can prevent the process of DN kidney damage.
  • the main clinical diagnosis is characterized by albumin excretion rate and albumin creatinine ratio.
  • Diabetic nephropathy is a modern medical disease name. There is no exact disease name corresponding to it in the ancient medical books. However, the literature records the long-term loss of diabetes, urinary edema, diarrhea, kidney elimination, and related diabetes. The clinical manifestations of kidney disease are very similar.
  • the carnosic acid is derived from the sage of the genus Lamiaceae, rosemary, sage, and sage, and has the effects of anti-oxidation, cardiovascular disease improvement, weight loss and lipid-lowering.
  • the present invention discloses a composition of carnosic acid for the preparation of a medicament for the treatment or prevention of diabetes.
  • the present invention discloses a composition of carnosic acid for the preparation of a medicament for treating or preventing diabetic nephropathy.
  • the present invention discloses the use of carnosic acid for the preparation of a composition for the treatment or prevention of type I or type II diabetes.
  • the present invention discloses a composition of carnosic acid for the preparation of cells that protect islet cells and repair damaged islet cells.
  • the present invention discloses a composition of carnosic acid for preparing kidney tissue for protecting a diabetic kidney patient, or for repairing damaged kidney tissue of a diabetic kidney patient.
  • the present invention discloses the use of carnosic acid for the preparation of a composition for treating or preventing hyperlipidemia.
  • the present invention discloses a composition of carnosic acid for the preparation of a medicament for treating or preventing nonalcoholic fatty liver.
  • the carnosic acid composition disclosed by the present invention is:
  • the pharmaceutically acceptable salt of carnosic acid includes a basic salt such as a sodium salt, a potassium salt and a calcium salt;
  • the composition is made up of a drug, a health supplement, or a functional food, and the excipient or carrier is an excipient or carrier commonly used in the pharmaceutical or food field, such as a diluent, a disintegrant, a lubricant, and the like.
  • the present invention discloses a rosemary petroleum ether extract for use in the preparation of a composition for treating or preventing any one or more of hyperlipidemia, hyperglycemia, nonalcoholic fatty liver disease, diabetic nephropathy and/or type 2 diabetes.
  • the present invention discloses a composition for preparing a rosemary petroleum ether extract, which is:
  • compositions for treating or preventing type I or type II diabetes For the preparation of a composition for treating or preventing type I or type II diabetes; or;
  • compositions for protecting islet cells, repairing damaged islet cells For the preparation of a composition for protecting islet cells, repairing damaged islet cells; or;
  • the invention discloses a composition for preparing a rosemary petroleum ether extract, wherein the components of the rosemary petroleum ether extract are: carnosic acid, carnosol, rosmarin, rosmarinol, Epidermal rosmarinol, methyl myristate, Salviol, 1-Phenanthrenecarboxylic acid, 2-Phenanthrenemethanol.
  • the invention discloses a composition for preparing a rosemary petroleum ether extract, wherein the components of the rosemary petroleum ether extract are: rosemary phenol, carnosol, carnosic acid, methyl rat tail acid, Salviol; its relative mass spectral peak area ratio: 2.23: 10.12: 39.64: 2.93: 32.32.
  • the invention discloses a composition for preparing a rosemary petroleum ether extract, which is used for preparing a composition for preventing and treating hyperlipidemia, preventing and treating hyperglycemia, or nonalcoholic fatty liver, diabetic nephropathy and type 2 diabetes.
  • the content of the rosemary petroleum ether extract ranges from 1 mg to 2 g.
  • the invention discloses a composition for preparing a rosemary petroleum ether extract, and the preparation method of the rosemary petroleum ether extract is:
  • the rosemary medicinal material is extracted with alcohol, the alcohol extract is dissolved in water, the aqueous solution is extracted with petroleum ether, the petroleum ether layer is combined, and the petroleum ether is recovered to obtain the rosemary petroleum ether portion.
  • the invention discloses a composition for preparing a rosemary petroleum ether extract, and the preparation method of the rosemary petroleum ether extract is:
  • Rosemary herbs are extracted with ethanol, combined with 95% ethanol extract, ethanol is recovered, the extract is added with water to dissolve, petroleum ether, petroleum ether and solution are added several times, and the petroleum ether layer is combined to recover petroleum ether to obtain rosemary petroleum ether. Part.
  • Rosemary herbs are extracted with methanol, combined with methanol extract, methanol is recovered, extract is added with water to dissolve, petroleum ether, petroleum ether and solution are added as many times as possible, and petroleum ether layer is combined to recover petroleum ether to obtain rosemary petroleum ether. .
  • the rosemary medicinal material is extracted with n-butanol, combined with n-butanol extract, n-butanol is recovered, the extract is added with water to dissolve, and petroleum ether, petroleum ether and solution are extracted several times, and the petroleum ether layer is combined to recover petroleum ether. Rosemary petroleum ether site.
  • the invention discloses a composition for preparing a rosemary petroleum ether extract, which is prepared into a medicine, a health care product, or a functional food, and the excipient or carrier is an excipient commonly used in the pharmaceutical or food field or A carrier such as a diluent, a disintegrant, a lubricant or the like.
  • the invention discloses a rosemary petroleum ether extract for preventing and treating hyperlipidemia, and the rosemary petroleum ether extract ranges from 1 mg to 2 g;
  • the present invention discloses a rosemary petroleum ether extract for controlling hyperglycemia, and the rosemary petroleum ether extract ranges from 1 mg to 2 g;
  • the invention discloses a rosemary petroleum ether extract for preventing and treating type 2 diabetes, and the rosemary petroleum ether extract ranges from 1 mg to 2 g;
  • the present invention discloses a composition for controlling non-alcoholic fatty liver, and the rosemary petroleum ether extract ranges from 1 mg to 2 g.
  • the invention discloses a rosemary petroleum ether extract for preventing and treating hyperlipidemia, and the rosemary petroleum ether extract is used in an amount ranging from 1 mg/day to 1 g/day;
  • the invention discloses a rosemary petroleum ether extract for controlling hyperglycemia, wherein the rosemary petroleum ether extract is used in an amount ranging from 1 mg/day to 1 g/day;
  • the invention discloses a rosemary petroleum ether extract for preventing and treating type 2 diabetes, and the rosemary petroleum ether extract is used in an amount ranging from 1 mg/day to 1 g/day;
  • the invention discloses a rosemary petroleum ether extract for controlling non-alcoholic fatty liver, and the rosemary petroleum ether extract is used in an amount ranging from 1 mg/day to 1 g/day.
  • the invention discloses a rosemary petroleum ether extract for preparing and preventing diabetic nephropathy, and the content of the rosemary petroleum ether extract ranges from 1 mg to 2 g.
  • FIG. 1 HE staining of kidney tissue
  • ITT Insulin tolerance curve
  • Figure 8 HE staining of liver pathological sections of a lipid-induced mouse model of diabetes in a high fat-induced mouse
  • Figure 10 Effect of C57BL/6 group, STZ+VEHICLE group, STZ+Irbesartan group and STZ+PFR group on urinary albumin excretion rate in DN mice
  • Figure 1 shows the effects of carnosic acid on pancreatic tissue of type I diabetic mice through the upper, middle and lower parts;
  • Figure 1 shows the normal group of mouse pancreatic tissue
  • Figure 1 shows the pancreatic tissue of the STZ group of mice
  • Figure 1 shows pancreatic tissue of STZ + carnosic acid group mice.
  • Figure 2 (Detailed view of HE staining of kidney tissue)
  • Figure 1 shows renal HE staining through the upper, middle and lower parts
  • Figure 1 shows the kidney tissue of the normal group of mice
  • Figure 1 shows the kidney tissue of the STZ group of mice
  • Figure 1 shows the kidney tissue of STZ + carnosic acid mice.
  • FIG. 1 shows renal PAS staining through the upper, middle and lower parts
  • Figure 2 shows the kidney tissue of the normal group of mice
  • the middle part of Figure 2 shows the kidney tissue of the STZ group of mice
  • FIG. 2 shows the kidney tissue of the STZ + carnosic acid group of mice.
  • Table 4 Effect of carnosic acid on blood glucose in STZ modeled diabetic nephropathy mice
  • Table 7 Effect of carnosic acid on albumin creatinine ratio in diabetic nephropathy mice.
  • mice were randomly divided into 3 groups, normal control group (10) and STZ group (20).
  • STZ group received continuous intraperitoneal injection of STZ (50mg/Kg) for 5 days. After two weeks, fasting blood glucose was measured. The blood glucose was greater than or equal to 13.8mmol/L.
  • the STZ group was randomly divided into model group and carnosic acid treatment group.
  • the oxalic acid treatment group was treated with oral gavage (15 mg/kg).
  • the STZ model group and the normal control group were treated with an equal volume of 0.5% CMCNa solution. After 10 weeks, the mice were fasted for 4 hours and the blood glucose was measured. As shown in Figure 1, there is a significant statistical difference.
  • the pathological results of the pancreas are shown in Figure 1.
  • the number of islets in the model group was significantly reduced compared with the normal control group.
  • the number of islets in the drug-administered group was increased to some extent compared with the model group, and the islet tissue morphology was restored to some extent, in order to better evaluate the carnosic acid.
  • the number of islets in each field was randomly selected from the HE stained pathological sections of pancreas. The results are shown in Table 2.
  • the total islet area in the corresponding field of view is shown in Table 3.
  • the drug-administered group can To some extent, the damage of STZ to the pancreas was repaired, the number of islets was significantly increased, and the total area of islets was significantly increased in each field of view.
  • carnosic acid can effectively repair islet injury and effectively reduce the blood sugar lowering caused by STZ. It can be speculated that the mechanism of carnosic acid treatment of type I diabetes may be effective in repairing damaged islet cells.
  • C57BL mice were randomly divided into 3 groups, normal control group (10 rats) and STZ group (20 rats).
  • STZ group received continuous intraperitoneal injection of STZ (50mg/Kg) for 5 days. After two weeks, fasting blood glucose was measured. The blood glucose was greater than or equal to 13.8 mM.
  • the STZ group was randomly divided into STZ group and carnosic acid treatment group. Only the STZ group and the normal oxalic acid treatment group were given oral gavage (30 mg/kg), the STZ group and the normal group were given the same volume of CMCNa, and after 12 weeks, the mice were placed in the mouse metabolic cage, free diet. Drinking water, using urine for analysis, the measured indicators are all evaluating the classic indicators of renal function.
  • the STZ group shown in Tables 4, 5 and 6 showed a significant increase of more than 10 times, indicating that the diabetic mice had severe damage to the kidney function.
  • the STZ group showed normal organ damage compared with the normal group. obvious. Based on the above table, the successful model of diabetic nephropathy can be obtained.
  • the present invention performs PAS staining on the kidney. And HE staining.
  • Figures 2 and 3 show that carnosic acid can effectively repair the organ damage of diabetic nephropathy, as shown in the STZ group, the glomerular mesangial area is widened, the matrix is increased, the glomerular basement membrane is thickened, and the renal tubule base is Membrane thickening and division, showing a typical characteristic of diabetic nephropathy, showing good modeling, while carnosic acid is repaired to a certain extent, the mesangial area is relatively reduced, and the stromal hyperplasia and tubular basement membrane thickening are improved, indicating treatment The effect is obvious.
  • the mobile phase is (A) 0.1% aqueous formic acid and (B) acetonitrile.
  • the mobile phase elution procedure is:
  • the flow rate is 0.8ml/min.
  • Mass spectrometry drying gas temperature 350 ° C, dry gas flow rate 10 L / h, atomization pressure 35 psig, ESI ion source, electrospray voltage 3500v, cracking voltage 120v, skimmer 65v, OCTIRF Vgp750v, scanning ion range 100-1100.
  • the components of the extract of rosemary petroleum ether are: carnosic acid, carnosol, rosmarin, rosmarin, epirubic rosin, methyl methyl tail. , Salviol, 1-Phenanthrenecarboxylic acid, 2-Phenanthrenemethanol and other compounds.
  • mice C57BL mice, SPF grade, male, body weight (20 ⁇ 2) g, were randomly divided into two groups, the first group of 10, the normal group, given normal feed, the remaining mice (40) were divided into three groups, Give high-fat diet (basic feed with 20% lard, 1.25% cholesterol, 0.5% sodium cholate) for free feeding and drinking.
  • the matrix, lovastatin group (60 mg/kg), the low concentration drug (75 mg/kg) of the petroleum ether fraction of Example 3, and the high concentration group (150 mg/kg) of the petroleum ether fraction of Example 3 were respectively administered. After 7 weeks of continuous feeding, blood was collected, sacrificed, and liver tissue was taken.
  • the experimental animals were kept in the SPF animal room, 12h light 12h night, free diet and drinking water, the animal state was normal during the experiment.
  • TC total cholesterol
  • HDL-C high density lipoprotein
  • LDL-C lower low density lipoprotein
  • TC Total cholesterol
  • TG triglyceride
  • HDL-C high-density lipoprotein
  • LDL-C low-density lipoprotein
  • HDL-C is mainly used to transport cholesterol from the blood to the liver for metabolism. It is therefore called “good” cholesterol, anti-atherosclerotic cholesterol, and the HDL-C is significantly elevated due to the increase in TC. Both the statin group and the drug-administered group significantly increased the content of this "good cholesterol", indicating that the treatment effect was remarkable.
  • the rosemary petroleum ether site can effectively improve hyperlipidemia, reduce TG, TC, LDL-C, and increase the content of "good cholesterol” HDL-C.
  • mice were randomly divided into 5 groups, 10 in each group, normal control group (10), model group. (10), low concentration group of rosemary petroleum ether (10), Example 3 high concentration group of rosemary petroleum ether, Example 3 low concentration drug of rosemary petroleum ether (75 mg/kg), examples 3
  • the high concentration group of petroleum ether (150mg/kg) except the normal control, the other groups were made with high-fat diet (basic feed added 20% lard, 1.25% cholesterol, 0.5% sodium cholate) for 6 weeks. Fasting overnight measurements, fasting blood glucose, glucose tolerance, and insulin tolerance after the end of dosing.
  • Glucose tolerance test method mice were fasted overnight. After oral administration of blood glucose 2g/kg, the mice were tested for blood glucose at 0, 15, 30, 60, 90, and 120 minutes, and the corresponding line area was calculated using GraphPad Prism software. (AUC).
  • Insulin tolerance test method mice were fasted overnight, and the mice were intraperitoneally injected with insulin 0.75 IU/kg at 0, 30, 60, 90, 120, and 150 min, respectively, and blood glucose was measured at the tip of the tail, and the corresponding values were calculated using GraphPad Prism software. Underline area (AUC).
  • the blood glucose level of the model group was about 1 times higher than that of the normal control group, indicating that the model was good, the blood glucose was decreased after administration, and the high concentration had a better hypoglycemic effect than the low concentration, and there was a statistical difference.
  • type 2 diabetes is characterized by insulin resistance, the body is not sensitive to insulin, and is commonly evaluated by glucose tolerance and insulin tolerance.
  • the Glucose Tolerance Test is an oral glucose load test used to understand the body's ability to regulate blood glucose after eating glucose. Through the glucose tolerance test, sugar metabolism abnormalities can be found early, which is currently recognized as the gold standard for diagnosis of diabetes.
  • OGTT can be used for differential diagnosis in order to determine whether or not diabetes is present.
  • the body has a mechanism to maintain blood sugar, oral glucose, rapid rise in blood glucose after a brief rise, that is, normal glucose tolerance, the corresponding area under the oral glucose tolerance curve is small, the use of sugar in diabetic patients, oral glucose After the blood glucose rises rapidly, the blood sugar declines slowly, that is, the corresponding under-line area of glucose tolerance decreases.
  • the glucose tolerance decreases, and the blood glucose state of the body after the fasting is simulated. The blood glucose of the model group at each time point is shown. Both were higher than the normal group, indicating that the modeling was successful.
  • the model showed that the glucose tolerance curves of the rosemary petroleum ether treatment group and the normal group were under the model group curve. In order to better quantify this phenomenon, the GraphPad Prism software was used to calculate.
  • the corresponding under-line area (AUC), the corresponding under-line area is also lower than the model group, and has statistical significance. It shows that the overall hypoglycemic effect of rosemary petroleum ether is obvious.
  • the insulin tolerance test is an experiment that reflects the body's sensitivity to insulin. It mainly simulates the change of sugar level after insulin secretion. After high fat modeling, the blood glucose level of each point after injection of the same amount of insulin is higher than that of the normal group. The corresponding under-line area was also higher than that of the normal group. The blood glucose level at each time point after treatment with rosemary petroleum ether was lower than that of the model group, and the corresponding under-line area (Table 8.) was lower than that of the high-fat-induced model group. The sugar effect is obvious. The results showed that the rosemary petroleum ether site can effectively increase insulin sensitivity and improve diabetes.
  • mice C57BL mice, SPF grade, male, body weight (20 ⁇ 2) g, were randomly divided into two groups, the first group of 10, the normal group, given normal feed, the remaining mice (40) were given high fat Feed (basic feed added 20% lard, 1.25% cholesterol, 0.5% sodium cholate) free to eat and drink. Divided into high fat group, high fat + lovastatin group (60mg / kg), high fat + Example 1 rosemary petroleum ether low concentration group (75mg / kg), high fat + Example 1 rosemary oil High concentration group of ether (150mg/kg). 7 weeks of observation:
  • liver function related indicators plasma ALT
  • Liver lipid accumulation related indicators liver weight, liver TG, liver TC administration, determination of mice, blood lipid levels. Statistical analysis was performed using a t test.
  • mice After modeling, the blood levels of TC, TG, HDL-C and LDL-C in the mice were significantly increased, and there were significant statistical differences, indicating successful modeling, and significant concentrations of TC, TG and LDL-C were administered. The reduction in dependence has significant statistical differences, indicating a significant therapeutic effect.
  • HDL-C is mainly used to transport cholesterol from the blood to the liver for metabolism. It is therefore called “good” cholesterol, anti-atherosclerotic cholesterol, and the HDL-C is significantly elevated due to the increase in TC. Both the statin group and the drug-administered group significantly increased the content of this "good cholesterol", indicating that the treatment effect was remarkable.
  • Alanine aminotransferase also known as alanine aminotransferase, abbreviated as GPT, ALT, alanine aminotransferase is mainly present in the liver cell cytoplasm, and its intracellular concentration is 1000-3000 times higher than serum. Hepatocytes raise the alanine aminotransferase in the blood. 1% of liver cell damage can double the concentration of alanine aminotransferase in the blood. Therefore, alanine aminotransferase is the most sensitive indicator of liver cell damage, the most clinically Commonly used indicators of liver function tests. After the mouse model was established, the ALT increased about 1.5 times, and the ALT administration returned to normal.
  • the pathological results showed that the fat accumulation in the liver of the mice was significant after the model establishment, the hepatic cell steatosis was obvious, and the fibrosis was increased, indicating non-alcoholic fat. Liver modeling was established, and lovastatin and rosemary petroleum ether were given, and pathological damage was significantly reversed. In addition, the liver weight increased significantly after high-fat modeling, and some degree of reversal after administration. Combined with blood ALT and TC, TG, LDL indicators indicate liver development toward hardening trend, liver weight in high-dose group gradually recovered, and statistically significant.
  • the data showed that non-alcoholic fatty liver model establishment was established, and the therapeutic effect after administration was significant, with statistical differences.
  • Triglyceride content in the liver, unit mg cholesterol / g fresh liver tissue
  • Diabetic nephropathy Diabetic nephropathy
  • Micro-albumin (m-Alb) test kit (ELISA method) Nanjing Institute of Built Biologicals Batch number: 20150526
  • STZ streptozotocin
  • 2.1 g of citric acid is added to 100 mL of double distilled water to prepare a liquid A.
  • 2.94 g of sodium citrate was added to 100 mL of double distilled water to prepare a liquid B.
  • the A and B liquids are mixed at 1:1.32, the pH value is measured by a pH meter, and the pH is adjusted to 4.2-4.5, that is, the citric acid buffer solution of the STZ is required.
  • STZ was prepared into a 3 mg/ml streptozotocin solution in buffer, protected from light, and injected within 30 minutes.
  • Drug formulation PFR was formulated into a solution of 7.72 mg/ml with 0.5% sodium carboxymethylcellulose (CMC-Na). Irbesartan was formulated into a 0.75 mg/ml solution with 0.5% CMC-Na.
  • mice Thirty-eight C57BL/6 mice were randomly divided into normal group (12 rats) and model group (26 rats) after 7 days of adaptive feeding, and fed a normal diet daily. After 7 days, the model group was intraperitoneally injected with STZ (60 mg/kg). After STZ5d was continuously administered, the tail blood was taken from the tail of the mouse to measure fasting blood glucose with a blood glucose meter. Mice with fasting blood glucose > 16 mmol/L were included in diabetic nephropathy animals.
  • mice Molded diabetic mice were randomly divided into 3 groups according to blood glucose and body weight, namely: STZ+Vehicle group (10) Only), STZ+PFR (154.4mg/kg) group (5), STZ+Irbesartan group (9). Another 12 normal mice were selected as the C57BL/6 group.
  • the specific dosage regimen is:
  • Each group of animals was intragastrically administered daily according to the above protocol for 24 hours.
  • urine was collected.
  • the mice were placed in a metabolic cage for 24 hours, and the water was not fasted and 24 hours of urine was collected.
  • the rats were fasted for 12 hours before the end of the experiment, and the fasting blood glucose was measured with a blood glucose meter.
  • mice The general state, fasting blood glucose, body weight, food intake and water intake of the mice in each group were monitored during the experiment.
  • mice in each group were collected for 24 hours urine samples one week before the experiment. They were not fasted during the period of urine retention. The mice were placed in a washed metabolic cage. After recording the urine volume, take 2 ml, centrifuge at 5000 rpm for 1 min, remove the sediment, and dispense. Store in an EP tube at -80 ° C in a refrigerator. Urine albumin concentration was measured using an ELISA kit.
  • the urine samples of each group of mice were collected for one week before the experiment, and the urine creatinine concentration was measured with a kit.
  • the general state showed: reduced activity; polydipsia, the average drinking water was three times that of C57BL/6 group; polyuria, litter was obviously moist, and needed to be replaced 1-2 times a day.
  • the weight gradually decreased, the spirit was wilting, and the coat color was gray and rough.
  • the STZ+PFR group and the STZ+Irbesartan group had improved water intake, urine output, weight loss, and mental status.
  • the blood glucose levels of the STZ+Vehicle group were significantly increased after 24 weeks (P ⁇ 0.001).
  • the blood glucose levels in the STZ+PFR group were significantly lower (P ⁇ 0.05), and the blood glucose levels in the STZ+Irbesartanl group were not statistically significant (P>0.05).
  • the urinary albumin excretion rate (UAER) in the STZ+VEHICLE group was significantly higher than that in the C57BL/6 group (P ⁇ 0.001). Compared with the STZ+VEHICLE group, the UAER in the STZ+PFR group was significantly lower (P ⁇ 0.01). There was no significant difference in UAER between STZ+Irbesartanl group (P>0.05).
  • the ratio of urinary albumin to creatinine (UAlb/UCr) in the STZ+VEHICLE group was significantly higher (P ⁇ 0.001).
  • the UAlb/UCr values in the STZ+PFR group were significantly lower (P ⁇ 0.01).
  • the UAlb/UCr values in the STZ+Irbesartanl group were not statistically significant (P>0.05).
  • Rosemary medicinal herbs 10 times the amount of methanol extraction 3 times, 1h / time, combined with methanol extract, methanol recovery, extract with water to dissolve, add petroleum ether, petroleum ether and aqueous solution according to 1; 1 extraction three times, combined with petroleum ether layer, The petroleum ether is recovered from the petroleum ether.
  • Rosemary medicinal material 10 times the amount of n-butanol extraction 3 times, 1h / time, combined with n-butanol extract, recovery of n-butanol, extract with water to dissolve, add petroleum ether, petroleum ether and aqueous solution according to 1; 1 extraction three times The petroleum ether layer is combined and the petroleum ether is recovered to obtain the rosemary petroleum ether portion.
  • Preparation process take the sage acid over 100 mesh sieve, add starch, magnesium stearate, mix well, make granules, dry, tablet, and get.
  • Preparation process take carnosic acid over 100 mesh sieve, add starch, magnesium stearate, mix well, make granules, dry, capsule, and get.
  • Dry yeast 5g warm water 90ml, water a little, flour 150g, carnosic acid 5mg, vegetable oil 10g, low sodium salt
  • Biscuits Sprinkle the yeast in warm water, stir and dissolve, and place the carnosic acid. Add flour and stir, add vegetable oil, and knead into a smooth dough; the dough is made into a 0.2 cm thick sheet. Press out the shape, puncture the hole, sprinkle water on the surface, sprinkle with a little low sodium salt, ferment for 10 minutes at room temperature; preheat the oven to 120 degrees, put it on the upper layer, and bake for about 10 minutes to get the food containing carnosic acid.
  • Preparation process the rosemary petroleum ether part of Example 3 is passed through a 100 mesh sieve, and starch and magnesium stearate are uniformly mixed to form granules, dried, and tableted.
  • Preparation process the rosemary petroleum ether part of Example 3 is passed through a 100 mesh sieve, and starch and magnesium stearate are uniformly mixed to form granules, dried, and encapsulated.
  • Preparation process Take the rosemary petroleum ether part of Example 3, add soybean soft phospholipid, colloid mill and mix, vacuum and press, that is, soft capsule.
  • Dry yeast 5g warm water 90ml, water a little, flour 150g, rosemary petroleum ether part 5mg, vegetable oil 10g, low sodium salt a little
  • Biscuit method The yeast was sprinkled in warm water and stirred to dissolve, and the rosemary petroleum ether portion of Example 3 was added. Add flour, stir, add vegetable oil, knead into smooth dough; dough into 0.2cm thick slices; press out the shape, puncture the hole, sprinkle water on the surface, sprinkle with a little low sodium salt, ferment at room temperature for 10 minutes; oven preheat At 120 degrees, it is placed on the upper layer and baked for about 10 minutes to obtain a food containing rosemary petroleum ether.

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Abstract

本发明公开了鼠尾草酸在制备用于预防或治疗糖尿病、糖尿病肾病、高脂血症、非酒精性脂肪肝的组合物中的用途，以及迷迭香石油醚提取物在制备用于预防或治疗高脂血症、高血糖、非酒精性脂肪肝、糖尿病肾病和/或2型糖尿病的组合物中的用途。

Description

鼠尾草酸的医药用途 技术领域
本发明公开了天然药物领域，公开了一种鼠尾草酸医药用途，本发明公开了鼠尾草酸预防或治疗I型糖尿病和/或糖尿病肾病的用途，迷迭香石油醚提取物预防或治疗高脂血症、高血糖、非酒精性脂肪肝、糖尿病肾病和/或2型糖尿病的用途。
背景技术
糖尿病是一种由于胰岛素分泌缺陷及(或)其生物学作用障碍引起的以高血糖为特征的代谢疾病，是在遗传基础上，有环境因素参与的遗传易感性疾病，是一种慢性全身性代谢疾病，严重危害人类健康，积极开展糖尿病的防治工作已成为主要的社会公共卫生问题。
I型糖尿病的发病原因主要是由于胰岛损伤胰岛素分泌绝对缺少，II型糖尿病是从胰岛素抵抗为主伴胰岛素相对不足到胰岛素分泌不足为主伴胰岛素抵抗的病理过程。目前公认II型糖尿病是一种在环境因素、生活方式的改变的作用下由多个基因分别或相互作用所导致的复杂遗传病，但是II型糖尿病的病因尚未完全阐明。
西医目前常采用饮食、运动疗法磺酰脲类、双胍类、噻唑烷二酮类、α-葡萄糖苷酶抑制剂、瑞格列奈、胰岛素等来治疗II型糖尿病，仅噻唑烷二酮类可改善胰岛素抵抗。病人在用西药治疗的同时，又不断地出现动脉硬化、冠心病和高血压等并发症，II型糖尿病的复杂机制及其导致的全身病变是西药治疗的薄弱环节。不论是磺脉类的促泌剂还是噻唑烷二酮类的增敏剂在实验和临床研究中都不具备明显的减肥功效，而肥胖是2型糖尿病乃至代谢综合征的重要病理基础。
II型糖尿病不仅是糖代谢紊乱疾病，还是脂肪代谢紊乱疾病。肥胖引起的II型糖尿病患者通常都有高糖和高脂饮食，并常伴有高脂血症。有研究表明提示血糖“正常”的肥胖患者己有β细胞分泌功能的异常。但是，大部分的肥胖者并不发展为糖尿病，说明机体的自身状况起重要作用。在同样的毒性作用下,易感人群很容易发展成II型糖尿病，而不敏感人群可能终身不发病，或者延迟发病。现代医学并不能够改变机体的易感性，但可以通过改变生活方式，减肥等手段来尽可能地减少早期的诱因，尽量的延长代偿期，从而延迟2型糖尿病的发生，甚至避免发病。
现代II型糖尿病的治疗观点已从已往的单纯控制血糖转为降糖、降脂、降 压、改善胰岛素抵抗等多环节治疗。但是，对于II型糖尿病患者，药量需逐渐增加，投药种类也常由单一用药逐渐变为联合用药，这不可避免地要考虑药物代谢对肝肾的副作用。故从传统中药中寻找低毒、疗效肯定的天然药物来治疗是目前研糖尿病究热点之一。
卫生部制定颁布的《中药新药治疗消渴病(糖尿病)的临床研究指导原则》中所制定的分类标准是目前采用最广泛的分类方法,即阴虚热盛证、气阴两虚证、阴阳两虚证和血疲气滞证四型。
糖尿病肾病(diabetic nephropathy，DN)是糖尿病最重要的并发症之一，我国的发病率亦呈上升趋势，目前已成为终末期肾病的第二位，仅次于肾小球肾炎，由于存在复杂的代谢紊乱，糖尿病发生肾脏损害，特别是一旦进入临床蛋白尿期，病情一般不可逆转，往往呈进行性发展直至终末期肾病，往往比其他肾脏疾病的治疗更加困难，是引起糖尿病患者死亡的主要原因之一，DN的发病机制和治疗药物的研制受到了医学界极大的重视。现代医学上多从饮食控制、血糖控制、降压、调整脂代谢等方面着手，或是采用透析、肾移植等治疗手段，尚无疗效确切的西药能阻止DN肾功能损害的进程。临床其主要诊断的特征为白蛋白排泄率，和白蛋白肌酐比值。
糖尿病肾病是现代医学病名，在古医籍中未有与之对应的确切病名，但文献记载的消渴病日久失治、误治后所出现尿独、水肿、肾消、关格等与糖尿病肾病的临床表现十分相似。如《圣济总录》曰:“消渴病久,肾气受伤,肾主水,肾气虚衰,气化失常,开阖不利,发为水肿。”，《杂病源流犀烛.三消源流》记载:“有消渴后身肿者,有消渴面目足膝肿小便少者。”，因而本病当属中医学中的“消渴”、“水肿”、“虚劳“等范畴。
对糖尿病肾病病机的认识，中医各家认识较为一致，多立足本虚标实、虚实夹杂，公开了肝脾肺肾诸脏，糖尿病肾病的病机特点是：以肾、脾、肝及气血阴阳之虚为本；标实以瘀血、水湿、痰浊最多。中医药在防治糖尿病肾病方面显示出独特的优势，从中药中发现改善临床症状、减轻蛋白尿、降低血糖、保护肾功能的天然产物是国内外研究的热点。
鼠尾草酸来源于唇形科鼠尾草、迷迭香、快乐鼠尾草、三叶鼠尾草等中，具有抗氧化、改善心血管疾病、减肥降脂等的功效。
至今未见有关鼠尾草酸治疗I糖尿病治疗或预防作用的报道。
至今未见有关鼠尾草酸的糖尿病肾病治疗或预防作用的报道。
至今未见有关迷迭香石油醚提取物治疗或预防高脂血症、高血糖、非酒精性脂肪肝、糖尿病肾病、2型糖尿病的报道。
发明内容
本发明公开了鼠尾草酸用于制备治疗或预防糖尿病的组合物。
本发明公开了鼠尾草酸用于制备治疗或预防糖尿病肾病的组合物。
本发明公开了鼠尾草酸用于制备治疗或预防I型或II型糖尿病的组合物。
本发明公开了鼠尾草酸用于制备保护胰岛细胞、修复受损的胰岛细胞的组合物。
本发明公开了鼠尾草酸用于制备保护糖尿病肾病人的肾脏组织、或、修复糖尿病肾病人的受损的肾脏组织的组合物。
本发明公开了鼠尾草酸用于制备治疗或预防高脂血症的组合物。
本发明公开了鼠尾草酸用于制备治疗或预防非酒精性脂肪肝的组合物。
本发明公开了的鼠尾草酸组合物,为：
为市售或按已知方法制备的鼠尾草酸或其药用盐、水合物或无水物；
其中鼠尾草酸药用盐包括碱式盐如钠盐、钾盐和钙盐；
组合物制成：药物、保健品、或功能性食品,赋形剂或载体为制药或食品领域中常用的赋形剂或载体，如稀释剂，崩解剂，润滑剂等。
本发明公开了迷迭香石油醚提取物用于制备治疗或预防高脂血症、高血糖、非酒精性脂肪肝、糖尿病肾病和/或2型糖尿病中任意一种或多种的组合物。
本发明公开了迷迭香石油醚提取物用于制备的组合物,为：
用于制备治疗或预防I型或II型糖尿病的组合物；或；
用于制备保护胰岛细胞、修复受损的胰岛细胞的组合物；或；
用于制备保护糖尿病肾病人的肾脏组织、或、修复糖尿病肾病人的受损的肾脏组织的组合物。
本发明公开了迷迭香石油醚提取物用于制备的组合物，迷迭香石油醚提取物的组分是：鼠尾草酸、鼠尾草酚、迷迭香酚、表迷迭香酚、表异迷迭香酚、鼠尾酸甲酯、Salviol、1-Phenanthrenecarboxylic acid、2-Phenanthrenemethanol。
本发明公开了迷迭香石油醚提取物用于制备的组合物，迷迭香石油醚提取物的组分是：迷迭香酚，鼠尾草酚，鼠尾草酸，鼠尾酸甲酯，Salviol；其相对质谱峰面积比值为：2.23：10.12：39.64：2.93：32.32。
本发明公开了迷迭香石油醚提取物用于制备的组合物，用于制备防治高脂血症、防治高血糖、或、非酒精性脂肪肝、糖尿病肾病、2型糖尿病的组合物，迷迭香石油醚提取物的含量范围是1毫克-2克。
本发明公开了迷迭香石油醚提取物用于制备的组合物，迷迭香石油醚提取物的制备方法是：
醇提取迷迭香药材，用水溶解醇提取物，用石油醚萃取水溶液，合并石油醚层，回收石油醚得迷迭香石油醚部位。
本发明公开了迷迭香石油醚提取物用于制备的组合物，迷迭香石油醚提取物的制备方法是：
迷迭香药材用乙醇提取，合并95％乙醇提取液，回收乙醇，浸膏加水使溶，加入石油醚，石油醚和溶液萃取多次，合并石油醚层，回收石油醚得迷迭香石油醚部位。
迷迭香药材用甲醇提取，合并甲醇提取液，回收甲醇，浸膏加水使溶，加入石油醚，石油醚和溶液按照萃取多次，合并石油醚层，回收石油醚得迷迭香石油醚部位。
迷迭香药材用正丁醇提取，合并正丁醇提取液，回收正丁醇，浸膏加水使溶，加入石油醚，石油醚和溶液萃取多次，合并石油醚层，回收石油醚得迷迭香石油醚部位。
本发明公开了迷迭香石油醚提取物用于制备的组合物，组合物制成：药物、保健品、或功能性食品,赋形剂或载体为制药或食品领域中常用的赋形剂或载体，如稀释剂，崩解剂，润滑剂等。
本发明公开了用于防治高脂血症的迷迭香石油醚提取物,迷迭香石油醚提取物的范围是1毫克-2克；
本发明公开了用于防治高血糖的迷迭香石油醚提取物,迷迭香石油醚提取物的范围是1毫克-2克；
本发明公开了用于防治2型糖尿病的迷迭香石油醚提取物,迷迭香石油醚提取物的范围是1毫克-2克；
本发明公开了用于防治非酒精性脂肪肝的组合物,迷迭香石油醚提取物的范围是1毫克-2克。
本发明公开了用于防治高脂血症的迷迭香石油醚提取物,迷迭香石油醚提取物的用量范围是1毫克/天-1克/天；
本发明公开了用于防治高血糖的迷迭香石油醚提取物,迷迭香石油醚提取物的用量范围是1毫克/天-1克/天；
本发明公开了用于防治2型糖尿病的迷迭香石油醚提取物,迷迭香石油醚提取物的用量范围是1毫克/天-1克/天；
本发明公开了用于防治非酒精性脂肪肝的迷迭香石油醚提取物,迷迭香石油醚提取物的用量范围是1毫克/天-1克/天。
本发明公开了用于制备防治糖尿病肾病的迷迭香石油醚提取物，迷迭香石油醚提取物的含量范围是1毫克-2克。
为了便于理解，下面通过附图和具体实施例对本发明的用途进行详细的描述。需要特别指出的是，具体实施例和附图仅是为了说明，显然本领域的技术人员可以根据本文说明，对本发明进行各种修正或改变，这些修正和改变也将 纳入本发明范围之内。
附图说明
图1.鼠尾草酸对胰岛细胞的修复
图2：肾脏组织HE染色图
图3：肾组织PAS染色
图4.迷迭香石油醚部位HPLC-ESI-Q-TOF-MS总离子流图
图5.迷迭香石油醚部位对高脂诱导的小鼠糖尿病模型口服糖耐量曲线(OGTT)
图6.迷迭香石油醚部位对高脂诱导的小鼠糖尿病模型胰岛素耐受曲线(ITT)
图7.迷迭香石油醚部位对高脂诱导的小鼠糖尿病模型肝脏病理切片PAS染色
图8.迷迭香石油醚部位对高脂诱导的小鼠糖尿病模型肝脏病理切片HE染色
图9.C57BL/6组、STZ+VEHICLE组、STZ+Irbesartan组和STZ+PFR组对DN小鼠血糖的影响
图10.C57BL/6组、STZ+VEHICLE组、STZ+Irbesartan组和STZ+PFR组对DN小鼠尿白蛋白排泄率的影响
图11.C57BL/6组、STZ+VEHICLE组、STZ+Irbesartan组和STZ+PFR组对DN小鼠UAlb/UCr的影响
图1的详细说明
图1通过上中下三个部分展示鼠尾草酸对I型糖尿病小鼠胰腺组织的影响；
图1的上部展示正常组小鼠胰腺组织；
图1的中部展示STZ组小鼠胰腺组织；
图1的下部展示STZ+鼠尾草酸组小鼠胰腺组织
图2(肾脏组织HE染色图)的详细说明
图1通过上中下三个部分展示肾脏HE染色；
图1的上部展示正常组小鼠肾脏组织；
图1的中部展示STZ组小鼠肾脏组织；
图1的下部展示STZ+鼠尾草酸小鼠肾脏组织。
图3(肾组织PAS染色)的详细说明
图2通过上中下三个部分展示肾脏PAS染色；
图2的上部展示正常组小鼠肾脏组织；
图2的中部展示STZ组小鼠肾脏组织；
图2的下部展示STZ+鼠尾草酸组小鼠肾脏组织。
本发明的数据记载于如下表格中:
表1.鼠尾草酸对STZ造模的I型糖尿病的10周后血糖的影响.
表2.胰岛数目统计分析
表3.每个视野下胰岛总面积统计分析
表4：鼠尾草酸对STZ造模的糖尿病肾病小鼠的血糖的影响；
表5：鼠尾草酸对糖尿病肾病小鼠的尿量的影响；
表6：鼠尾草酸对糖尿病肾病小鼠的UAE的影响；
表7：鼠尾草酸对糖尿病肾病小鼠的白蛋白肌酐比值的影响。
表8.迷迭香石油醚部位质谱表征
表9.血浆中甘油三酯含量
表10.血浆中总胆固醇含量
表11.血浆中高密度脂蛋白含量
表12.血浆中低密度脂蛋白含量
表13.禁食血糖水平
表14.口服糖耐量线下面积值
表15.胰岛素耐受线下面值
表16.血浆中ALT含量
表17.肝脏重量
表18.肝脏中总胆固醇含量
表19.肝脏中甘油三酯含量
表20.实验C56BL/6小鼠空腹血糖
表21实验C56BL/6小鼠尿白蛋白排泄率
表22.实验C56BL/6小鼠尿微量白蛋白与肌酐比值
具体实施方式
实施例1
为了研究鼠尾草酸对I型糖尿病血糖影响，本实验采用经典的I型糖尿病模型，使用C57BL小鼠随机分为3组，正常对照组(10只)，STZ组(20只)。STZ组连续腹腔注射STZ(50mg/Kg)5天，两周后测定禁食血糖，血糖大于等于13.8mmol/L为造模成功，将STZ组随机分为模型组和鼠尾草酸治疗组，鼠尾草酸治疗组采用口服灌胃(15mg/kg)，STZ模型组、正常对照组采用口服灌胃等体积的0.5％的CMCNa溶液，10周后，小鼠禁食4h后测定血糖，结果如表1所示，具有显著的统计学差异。
鼠尾草酸的配置方法:用0.5％的CMCNa溶液溶解.
动物的一般情况:
动物毛发，活动，精神状态等均无差别
表1.鼠尾草酸对STZ造模的I型糖尿病的10周后血糖的影响.
Figure PCTCN2016074780-appb-000001
注：*P<0.05，**P<0.01，***P<0.001，与STZ组相比。
如表1.所示STZ组相比正常组血糖升高了3倍多，提示造模很成功，鼠尾治疗10周后血糖显著降低
表2.胰岛数目统计分析
Figure PCTCN2016074780-appb-000002
表3.每个视野下胰岛总面积统计分析
Figure PCTCN2016074780-appb-000003
Figure PCTCN2016074780-appb-000004
胰腺病理结果如图1所示，模型组相比正常对照组胰岛数目明显减少，给药组胰岛数目相比模型组一定程度增加，胰岛组织形态在一定程度恢复，为了更好的评估鼠尾草酸对胰腺的修复作用，在胰腺HE染色病理切片中随机选取9个视野观察每个视野下胰岛数目结果如表2，测定相应视野下总的胰岛面积结果如表3所示，给药组能够在一定程度上修复STZ对胰腺的损伤，胰岛数目显著增加，每个视野下胰岛总面积显著增加。总之鼠尾草酸能够有效的修复胰岛损伤，有效的降低STZ引起的血糖降低。有此可以推测，鼠尾草酸治疗I型糖尿病的机制可能是有效的修复受损胰岛细胞。
实施例2
为了研究鼠尾草酸对糖尿病肾病血糖影响，使用C57BL小鼠随机分为3组，正常对照组(10只)，STZ组(20只)。STZ组连续腹腔注射STZ(50mg/Kg)5天，两周后测定禁食血糖，血糖大于等于13.8mM为造模成功，将STZ组随机分为STZ组和鼠尾草酸治疗组，每组10只，STZ组给予和正常鼠尾草酸治疗组采用口服灌胃(30mg/kg)，STZ组和正常组给予等体积的CMCNa，12周后，将小鼠放入小鼠代谢笼中，自由饮食饮水，将尿液用于分析，所测指标均评估肾功能的经典指标。
表4、5、6所示STZ组相比空白对照组，显著升高均在10倍以上，说明糖尿病小鼠肾功能出现严重损伤，如图2、3所示STZ组相比正常组器官损伤明显。综合以上表图可以得出糖尿病肾病造模成功。
表4.结果表明鼠尾草酸能够有效的降低STZ造成的血糖升高，逆转由糖尿病肾病引起的尿量增加(表5)，有效的降低糖尿病肾病引起的24h蛋白排泄率升高(表6)。临床上常用尿液中白蛋白和肌酐的比值评估肾功能，鼠尾草酸有效的降低糖尿肾病引起的白蛋白和肌酐的比值升高(表7)。尿量、24h蛋白排泄率白蛋白和肌酐的比值的结果表明鼠尾草酸治疗后很好的改善肾功能。
为了更进一步的评估肾脏器官性损伤的改变，本发明对肾脏进行PAS染色 和HE染色。图2，3显示：鼠尾草酸能够有效的修复糖尿病肾病的器官性损伤逆转，如图所示STZ组肾小球系膜区增宽，基质增加，肾小球基膜增厚，肾小管基膜增厚及分裂，呈现出极为典型的糖尿病肾病特征，显示造模良好，而鼠尾草酸在一定程度上修复，系膜区相对缩小，改善了基质增生以及肾小管基膜增厚，说明治疗作用明显效果显著。
表4.鼠尾草酸对STZ造模的糖尿病肾病的血糖的影响.
Figure PCTCN2016074780-appb-000005
注：*P<0.05，**P<0.01，***P<0.001，与STZ组相比。
表5.鼠尾草酸对STZ造模的糖尿病肾病的尿量的影响.
Figure PCTCN2016074780-appb-000006
注：*P<0.05，**P<0.01，***P<0.001，与STZ组相比。
表6.鼠尾草酸对STZ造模的糖尿病肾病的UAE(尿白蛋白排泄率)的影响.
Figure PCTCN2016074780-appb-000007
注：*P<0.05，**P<0.01，***P<0.001，与STZ组相比。
表7.鼠尾草酸对STZ造模的糖尿病肾病的UACR(尿白蛋白/肌酐比值)的影响.
Figure PCTCN2016074780-appb-000008
Figure PCTCN2016074780-appb-000009
注：*P<0.05，**P<0.01，***P<0.001，与STZ组相比。
实施例3
迷迭香石油醚部位提取工艺
迷迭香药材10倍量95％乙醇提取3次，1h/次，合并95％乙醇提取液，回收乙醇，浸膏加水使溶，加入石油醚，石油醚和溶液按照1；1萃取三次，合并石油醚层，回收石油醚得迷迭香石油醚部位。
实施例4
迷迭香石油部位成分鉴定
95％乙醇溶解迷迭香石油醚部位10mg/ml,12,000rpm离心10min，取上清进行液相分析，
采用Agilent 1290UPLC-6520 QTOF MS，
流动相为(A)0.1％甲酸水溶液和(B)乙腈.
流动相洗脱程序为:
10％B(0min),
30％B(5min),
50％B(10min),
80％B(15min),
100％B(25-30min).
流速为0.8ml/min.
质谱干燥气温度350℃，干燥气体流速10L/h，雾化气压35psig，ESI离子源，电喷雾电压3500v，裂解电压120v,skimmer 65v，OCTIRF Vgp750v，扫描离子范围100-1100.二级碰撞电压20v；40v；70v。
质谱鉴定结果如下表
能确定本发明公开了迷迭香石油醚提取物的组分是：鼠尾草酸、鼠尾草酚、迷迭香酚、表迷迭香酚、表异迷迭香酚、鼠尾酸甲酯、Salviol、1-Phenanthrenecarboxylic acid、2-Phenanthrenemethanol等化合物。
表8.迷迭香石油醚部位质谱表征
Figure PCTCN2016074780-appb-000010
Figure PCTCN2016074780-appb-000011
Figure PCTCN2016074780-appb-000012
Figure PCTCN2016074780-appb-000013
Figure PCTCN2016074780-appb-000014
Figure PCTCN2016074780-appb-000015
迷迭香酚，鼠尾草酚，鼠尾草酸，鼠尾酸甲酯，Salviol。其质谱相对峰面积比值为：2.23：10.12：39.64：2.93：32.32(图谱如图1.所示)。
实施例5
迷迭香石油醚部位对于高脂血动物模型的作用
1、实验动物及方法：
C57BL小鼠，SPF级，雄性，体重(20±2)g，随机分成两组，第一组10只，为正常组，给与正常饲料，其余小鼠(40只)分为三组组，给与高脂饲料(基础饲料加入20％猪油，1.25％胆固醇，0.5％的胆酸钠)自由摄食、饮水。分别给予基质、洛伐他汀组(60mg/kg)，实施例3石油醚部位低浓度药物(75mg/kg)、实施例3石油醚部位高浓度组(150mg/kg)。连续喂养7周后，采血、处死、取肝脏组织。
观察指标：
a动物的一般情况
实验动物在SPF动物房中饲养，12h光照12h黑夜，自由饮食饮水，实验期间动物状态正常。
c血脂相关指标：TC、TG、HDL、LDL
3、实验结果：
结果表明：实施例3迷迭香石油醚部位均可明显降低血浆中的总胆固醇(TC)和甘油三酯(TG)，升高高密度脂蛋白(HDL-C)，降低低密度脂蛋白(LDL-C)给药结束后测定小鼠、血脂水平。使用t检验进行统计学分析。结果显示，迷迭香石油醚部位对高脂改善作用效果明显，具有显著统计学差异。
表9.血浆中甘油三酯含量，单位：mM
Figure PCTCN2016074780-appb-000016
与高脂组比较，*P<0.05,**P<0.01,***P<0.001
表10.血浆中总胆固醇含量，单位：mM
Figure PCTCN2016074780-appb-000017
*P<0.05,**P<0.01,***P<0.001
表11.血浆中高密度脂蛋白含量，单位：mM
Figure PCTCN2016074780-appb-000018
与高脂组比较，*P<0.05,**P<0.01,***P<0.001
表12.血浆中低密度脂蛋白含量，单位：mM
Figure PCTCN2016074780-appb-000019
与高脂组比较，*P<0.05,**P<0.01,***P<0.001
造模后小鼠的血液中总胆固醇(TC)、甘油三酯(TG),高密度脂蛋白(HDL-C)、低密度脂蛋白(LDL-C)均显著升高，且具有显著的统计学差异，表明造模成功，给药TC,TG,LDL-C均显著性浓度依赖性的降低，具有显著的 统计学差异，说明治疗效果显著。
HDL-C主要作用为将胆固醇从血液运输至肝脏中代谢，因此被称为"好"胆固醇，抗动脉粥样硬化的胆固醇，由于TC的升高造模组HDL-C显著升高，洛伐他汀组和给药组均能显著性的升高这一“好胆固醇”的含量，说明治疗效果显著。总之迷迭香石油醚部位能够有效的改善高脂血症，降低TG,TC,LDL-C，升高“好胆固醇”HDL-C的含量。
实施例6
迷迭香石油醚部位对于高血糖动物模型的作用
为了研究迷迭香石油醚部位对糖尿病的影响，本实验采用经典高脂诱导小鼠糖尿病模型，使用C57BL小鼠随机分为5组，每组各10只，正常对照组(10)，模型组(10)，迷迭香石油醚部位低浓度组(10)，实施例3迷迭香石油醚部位高浓度组，实施例3迷迭香石油醚部位低浓度药物(75mg/kg)、实施例3石油醚部位高浓度组(150mg/kg)其中除正常对照外，其他组均采用高脂饮食(基础饲料加入20％猪油，1.25％胆固醇，0.5％的胆酸钠)造模6周，给药结束后禁食过夜测定、空腹血糖、葡萄糖耐量和胰岛素耐量。糖耐量实验方法：小鼠禁食过夜，小鼠口服血糖2g/kg后分别在0，15，30，60，90，120min时，尾尖采血测定相应血糖，使用GraphPad Prism软件计算相应线下面积(AUC)。
胰岛素耐受实验方法：小鼠禁食过夜，小鼠腹腔注射胰岛素0.75IU/kg后分别在0，30，60，90，120，150min时，尾尖采血测定相应血糖，使用GraphPad Prism软件计算相应线下面积(AUC)。
所示，模型组相比正常对照组血糖约升高1倍，说明造模良好，给药后血糖有所降低，且高浓度相比低浓度具有更好的降血糖作用，且具有统计学差异。由于II型糖尿病主要特征为胰岛素抵抗，即机体对胰岛素不敏感，常用糖耐量和胰岛素耐量实验评估。糖耐量试验是一种口服葡萄糖负荷试验，用于了解机体对进食葡萄糖后的血糖调节能力。通过糖耐量试验，可以早期发现糖代谢异常，是目前公认的诊断糖尿病的金标准，在血糖增高但尚未达到糖尿病诊断标准时，为明确是否患糖尿病，可以采用OGTT进行鉴别诊断。正常情况下，机体有一套维持血糖的机制，口服葡萄糖，血糖短暂升高后迅速恢复正常，即糖耐量正常，相应的口服糖耐量曲线线下面积较小，糖尿病患者糖的利用障碍，口服葡萄糖后血糖迅速升高，血糖下降速度较慢，即糖耐量减退相应的线下面积较大，该实验中糖耐量减退，模拟禁食后机体血糖状态，所示，模型组在各个时间点的血糖均高于正常组，说明造模很成功，模型组呈现迷迭香石油醚部位治疗组和正常组的糖耐量曲线均在模型组曲线下，为了更好的定量这一现象使用GraphPad Prism软件计算相应线下面积(AUC)，，相应的线下面积也均低于模型组，且具有统计意义。说明了迷迭香石油醚部位整体降糖效果明显。
胰岛素耐量试验是反映机体对胰岛素敏感性的实验，主要模拟的是机体分泌胰岛素后糖水平的变化，高脂造模后，注射相同量的胰岛素后各个点的血糖值均高于正常组，其相应的线下面积也高于正常组，迷迭香石油醚部位治疗后各个时间点的血糖数值低于模型组，相应线下面积(表8.)低于高脂诱导的模型组，说明降糖效果明显。结果表明迷迭香石油醚部位能够有效的增加胰岛素敏感性改善糖尿病。
为了初步探讨迷迭香石油醚部位除改善胰岛素抵抗外是否有其他的改善糖尿病的机制，我们进行了肝脏PAS染色的病理分析，PAS主要针对糖原染色，结果显示，造模后糖原减少即紫红色减少，给药后紫红色逐渐增加，说明糖原储备增加。由于肝脏中的糖原是由血中的葡萄糖合成而来。由此推测迷迭香石油醚部位可能通过增加血液中葡糖糖合成肝糖原而产生降糖作用。
以上数据表明迷迭香石油醚部位能够显著降低血液中葡萄糖含量(表13)显著提高糖耐受(表14)显著增加胰岛素敏感性(表15)。
表13.禁食血糖水平，单位：mM
Figure PCTCN2016074780-appb-000020
*P<0.05,**P<0.01,***P<0.001
表14.口服糖耐量线下面积值
Figure PCTCN2016074780-appb-000021
与高脂组，*P<0.05,**P<0.01,***P<0.001
表15.胰岛素耐受线下面值
Figure PCTCN2016074780-appb-000022
Figure PCTCN2016074780-appb-000023
与高脂组，*P<0.05,**P<0.01,***P<0.001
实施例7
迷迭香石油醚部位对高脂血症和非酒精性脂肪肝的治疗作用
实验动物及方法：
C57BL小鼠，SPF级，雄性，体重(20±2)g，随机分成两组，第一组10只，为正常组，给与正常饲料，其余小鼠(40只)分别给予给与高脂饲料(基础饲料加入20％猪油，1.25％胆固醇，0.5％的胆酸钠)自由摄食、饮水。分为高脂组，高脂+洛伐他汀组(60mg/kg)、高脂+实施例1迷迭香石油醚部位低浓度组(75mg/kg)、高脂+实施例1迷迭香石油醚部位高浓度组(150mg/kg)。给药7周观察指标：
a动物的一般情况
实验动物，12h光照12h黑夜，自由饮食饮水，实验期间动物状态正常。
b肝功能相关指标：血浆ALT
c血脂相关指标：TC、TG、HDL、LDL
d病理学检查：HE染色
e肝脏脂类蓄积相关指标：肝脏重量、肝脏中TG、肝脏中TC给药结束后测定小鼠、血脂水平。使用t检验进行统计分析。
2.结果分析
造模后小鼠的血液中TC，TG，HDL-C，LDL-C均显著升高，且具有显著的统计学差异，表明造模成功，给药TC，TG，LDL-C均显著性浓度依赖性的降低，具有显著的统计学差异，说明治疗效果显著。
HDL-C主要作用为将胆固醇从血液运输至肝脏中代谢，因此被称为"好"胆固醇，抗动脉粥样硬化的胆固醇，由于TC的升高造模组HDL-C显著升高，洛伐他汀组和给药组均能显著性的升高这一“好胆固醇”的含量，说明治疗效果显著。
丙氨酸转氨酶又称谷丙转氨酶，简称GPT、ALT,丙氨酸转氨酶主要存在于肝细胞浆内，其细胞内浓度高于血清中1000-3000倍。肝细胞会使血液中的丙氨酸转氨酶升高。1％的肝细胞损伤可以使血液中的丙氨酸转氨酶浓度升高一倍。因此，丙氨酸转氨酶是反映肝细胞受损程度最灵敏的指标，临床上是最为 常用的肝功能检查指标。小鼠造模后ALT约升高1.5倍，给药ALT基本恢复正常，如病理结果显示，造模后小鼠肝脏中脂肪蓄积显著，肝细胞脂肪变性明显，纤维化增加，说明非酒精性脂肪肝造模成立，给予洛伐他汀和迷迭香石油醚部位，病理损伤显著逆转。此外高脂造模后肝脏重量显著增加，给药后一定程度逆转)结合血液中ALT和TC，TG，LDL指标说明肝脏向硬化趋势发展，高剂量组肝脏重量逐渐恢复，且具有统计学差异。
数据显示肝脏中甘油三酯和胆固醇含量的结果显示，造模后肝脏中甘油三酯升高约1.7倍，胆固醇约升高3倍，说明肝脏中甘油三酯胆固醇蓄积显著，给予洛伐他汀和，迷迭香石油醚部位基本恢复正常。综上所述，数据显示非酒精性脂肪肝造模成立，给药后治疗效果显著，具有统计学差异。
表16.血浆中ALT含量，单位：IU/L
Figure PCTCN2016074780-appb-000024
与高脂组，*P<0.05,**P<0.01,***P<0.001
表17.肝脏重量，单位：g
Figure PCTCN2016074780-appb-000025
表18.肝脏中总胆固醇含量，单位：mg胆固醇/g新鲜肝脏组织
Figure PCTCN2016074780-appb-000026
Figure PCTCN2016074780-appb-000027
与高脂组，*P<0.05,**P<0.01,***P<0.001
表19.肝脏中甘油三酯含量，单位：mg胆固醇/g新鲜肝脏组织
Figure PCTCN2016074780-appb-000028
与高脂组，*P<0.05,**P<0.01,***P<0.001
实施例8
提取物对C57BL/6小鼠实验性糖尿病肾病的治疗作用实验研究
本实验所用英文缩写词如下。
英文缩写词表
英文缩写  英文全称                          中文全称
PFR     Petroleum ethPFR fraction of rosemary      迷迭香石油醚提取物
DN      Diabetic nephropathy                       糖尿病肾病
ESRD    End-stage renal diseas                     终末期肾病
FBG     Fasting blood glucose                      空腹血糖
STZ      Streptozocin                              链脲佐菌素
UAER    Urinary albumin excretion rate             尿白蛋白排泄率
UAlb     Urinary albumin                           尿微量白蛋白
UCr      Urinary creatinine                        尿肌酐
本发明的实验方法及实验结果如下
实验材料
实验动物
C57BL/6小鼠，雄性，体重20±2g，由扬州大学动物医学实验教学中心提供，质量合格证号：SCXK-(苏)2012-0004。
药品与试剂
迷迭香                   产地：安徽(由生药学李会军教授鉴定)
厄贝沙坦(Irbesartan)     日本TCI公司           批号：UIGAD-QM
链脲佐菌素(STZ)          美国阿拉丁工业公司     批号：G1324033
肌酐测定试剂盒           南京建成生物研究所     批号：20150814
微量白蛋白(m-Alb)测试盒(ELISA方法)  南京建成生物研究所   批号：20150526
以下将实施例3的迷迭香石油醚提取物简称为PFR。
实验仪器
HEA-230血糖仪                     欧姆龙集团
Centrifuge 5427R离心机            艾本德(Eppendorf)公司
BioTek SynPFRgy2酶标仪            美国伯腾仪器有限公司
QL-901 Vortex混悬器               海门市其林贝尔仪器制造
实验方法
药物配制
饮食配方：正常小鼠饲料。
链脲佐菌素(STZ)溶液的配制：STZ-20℃以下保存。临用时以柠檬酸2.1g加入双蒸水100mL中配成A液。柠檬酸钠2.94g加入双蒸水100mL中配成B液。用时将A、B液按1：1.32混合，pH计测定pH值，调节pH＝4.2-4.5，即是所需配置STZ的柠檬酸缓冲液。临用时将STZ用缓冲液配制成3mg/ml的链脲佐菌素溶液，避光，30min内注射完毕。
药物配制：将PFR用0.5％羧甲基纤维素钠(CMC-Na)配成7.72mg/ml的溶液。Irbesartan用0.5％的CMC-Na配成0.75mg/ml的溶液。
实验模型的建立及给药方法
将38只C57BL/6小鼠适应性喂养7d后随机分为正常组(12只)和模型组(26只),每日喂食正常饮食。7d后，模型组腹腔注射STZ(60mg/kg)。连续给予STZ5d后小鼠尾尖取血用血糖仪检测空腹血糖。将空腹血糖值＞16mmol/L的小鼠列入糖尿病肾病动物。
将成模的糖尿病小鼠按血糖和体重随机分为3组，即：STZ+Vehicle组(10 只)、STZ+PFR(154.4mg/kg)组(5只)、STZ+Irbesartan组(9只)。另选12只正常组小鼠作为C57BL/6组。具体的给药方案为：
C57BL/6组：               灌胃0.5％CMC-Na 20mL/kg
STZ+Vehicle组：            灌胃0.5％CMC-Na 20mL/kg
STZ+Irbesartan组：         灌胃Irbesartan 15mg/kg
STZ+PFR组：              灌胃PFR 154.4mg/kg
各组动物每日按照上述方案灌胃给药，连续24w。实验前一周，收集尿液。将小鼠放入代谢笼中24h，不禁食不禁水，收集24h尿液。实验结束前12h禁食不禁水，用血糖仪测空腹血糖。
观察指标及标本采集
一般指标的观察
监测实验过程中各组小鼠的一般状态、空腹血糖、体重、摄食及摄水量等变化。
尿蛋白的测定
实验前一周收集各组小鼠24h尿样，留尿期间不禁食，不禁水，将小鼠放入洗净的代谢笼内，记录尿量后取2ml，5000rpm离心1min，去除沉渣，分装于EP管中，-80℃冰箱保存。用ELISA试剂盒检测尿白蛋白浓度。
尿肌酐的测定
实验前一周收集各组小鼠24h尿样，用试剂盒检测尿肌酐浓度。
统计方法
实验数据，计量资料进行组间t检验，P＜0.05为具有显著性统计学意义。(*P＜0.05，**P＜0.01，***P＜0.001)
实验结果
一般状态观察
模型组小鼠腹腔注射STZ 14d后，一般状态表现为：活动减少；多饮，平均饮水量是C57BL/6组的三倍；多尿，垫料明显潮湿，每日需更换1-2次，体重逐渐下降，精神萎靡，毛色灰暗，粗糙。与STZ+Vehicle组比较，STZ+PFR组，STZ+Irbesartan组小鼠的饮水量、尿量增多，体重下降及精神状态等均有所改善。
PFR对的影响
与C57BL/6组比较，24周后，STZ+Vehicle组小鼠血糖值均显著升高(P＜0.001)。与STZ+VEHICLE组比较，STZ+PFR组血糖值均显著降低(P＜0.05)，STZ+Irbesartanl组血糖值无统计学意义(P＞0.05)。
表20.实验C56BL/6小鼠空腹血糖
Figure PCTCN2016074780-appb-000029
Figure PCTCN2016074780-appb-000030
PFR对DN小鼠尿白蛋白排泄率的影响
与C57BL/6组比较STZ+VEHICLE组尿白蛋白排泄率(UAER)均显著升高(P＜0.001)。与STZ+VEHICLE组比较，STZ+PFR组UAER显著降低(P＜0.01)。STZ+Irbesartanl组UAER无统计学意义(P＞0.05).。
表21实验C56BL/6小鼠尿白蛋白排泄率
Figure PCTCN2016074780-appb-000031
PFR对DN小鼠尿微量白蛋白与肌酐比值的影响
与C57BL/6组比较，STZ+VEHICLE组尿微量白蛋白与肌酐比值(UAlb/UCr)均显著升高(P＜0.001)。与STZ+VEHICLE组比较，STZ+PFR组UAlb/UCr值显著降低(P＜0.01)。STZ+Irbesartanl组UAlb/UCr值无统计学意义(P＞0.05)。
表22.实验C56BL/6小鼠尿微量白蛋白与肌酐比值
Figure PCTCN2016074780-appb-000032
实施例9
迷迭香石油醚部位提取工艺
迷迭香药材10倍量甲醇提取3次，1h/次，合并甲醇提取液，回收甲醇，浸膏加水使溶，加入石油醚，石油醚和水溶液按照1；1萃取三次，合并石油醚层，回收石油醚得迷迭香石油醚部位。
实施例10
迷迭香石油醚部位提取工艺
迷迭香药材10倍量正丁醇提取3次，1h/次，合并正丁醇提取液，回收正丁醇，浸膏加水使溶，加入石油醚，石油醚和水溶液按照1；1萃取三次，合并石油醚层，回收石油醚得迷迭香石油醚部位。
实施例11
鼠尾草酸的制剂
鼠尾草酸片剂
鼠尾草酸10mg,淀粉88g,硬脂酸镁3g
制备工艺：取鼠尾草酸过100目筛，加淀粉、硬脂酸镁混合均匀，制成颗粒，干燥，压片，即得。
鼠尾草酸胶囊
鼠尾草酸10mg,淀粉88g,硬脂酸镁3g
制备工艺：取鼠尾草酸过100目筛，加淀粉、硬脂酸镁混合均匀，制成颗粒，干燥，装胶囊，即得。
实施例12
鼠尾草酸的食品制剂
干酵母5g、温水90ml、水少许、面粉150g、鼠尾草酸5mg、植物油10g、低钠盐少许
饼干做法：把酵母撒在温水里搅拌倒溶化，放鼠尾草酸。加入面粉搅拌，再加入植物油，揉成光滑的面团；面团制成0.2cm厚的薄片。压出造型，刺洞，表面撒上水，撒上少许低钠盐，室温发酵10分钟；烤箱预热120度，放在上层，烤约10分钟，得到含鼠尾草酸的食品。
实施例13
迷迭香石油醚部位的制剂
迷迭香石油醚部位的片剂：迷迭香石油醚部位10mg，淀粉88g，硬脂酸镁3g
制备工艺：取实施例3的迷迭香石油醚部位过100目筛，加淀粉、硬脂酸镁混合均匀，制成颗粒，干燥，压片，即得。
迷迭香石油醚部位的胶囊
迷迭香石油醚部位10mg，淀粉88g，硬脂酸镁3g
制备工艺：取实施例3的迷迭香石油醚部位过100目筛，加淀粉、硬脂酸镁混合均匀，制成颗粒，干燥，装胶囊，即得。
迷迭香石油醚部位的软胶囊
迷迭香石油醚部位10mg，大豆卵磷脂100g
制备工艺：取实施例3的迷迭香石油醚部位，加大豆软磷脂，胶体磨混匀，抽真空，压制，即得软胶囊。
迷迭香石油醚部位的冻干粉：
迷迭香石油醚部位2.0g，亚硫酸钠4.0g，乙醇50ml，加水定容至1000mL；
制备工艺：取实施例3的迷迷迭香石油醚部位分散在乙醇中，亚硫酸钠溶 于水中，在超声或搅拌条件下将拿硫酸钠溶液逐渐加入，使成澄清透明溶液；补加水定容至足量；经0.22μM微孔滤膜过滤，冷冻干燥得到。
实施例14
迷迭香石油醚部位的食品制剂
干酵母5g、温水90ml、水少许、面粉150g、迷迭香石油醚部位5mg，植物油10g、低钠盐少许
饼干做法：把酵母撒在温水里搅拌倒溶化，加入实施例3的迷迭香石油醚部位。加入面粉搅拌，再加入植物油，揉成光滑的面团；面团制成0.2cm厚的薄片；压出造型，刺洞，表面撒上水，撒上一点低钠盐，室温发酵10分钟；烤箱预热120度，放在上层，烤约10分钟，得含迷迭香石油醚部位的食品。

Claims (14)

  1. 鼠尾草酸用于制备治疗或预防糖尿病的组合物。
  2. 鼠尾草酸用于制备治疗或预防糖尿病肾病的组合物。
  3. 根据权利要求1-2中的组合物,其特征为：用于制备治疗或预防I型或II型糖尿病的组合或，用于制备保护胰岛细胞、修复受损的胰岛细胞的组合物，或，用于制备保护糖尿病肾病人的肾脏组织、或、修复糖尿病肾病人的受损的肾脏组织的组合物。
  4. 鼠尾草酸用于制备治疗或预防高脂血症的组合物。
  5. 鼠尾草酸用于制备治疗或预防非酒精性脂肪肝的组合物。
  6. 根据权利要求1-2中的组合物,其特征为：
    为市售或按已知方法制备的鼠尾草酸或其药用盐、水合物或无水物；
    其中鼠尾草酸药用盐包括碱式盐如钠盐、钾盐和钙盐；
    组合物制成：药物、保健品、或功能性食品,赋形剂或载体为制药或食品领域中常用的赋形剂或载体，如稀释剂，崩解剂，润滑剂等。
  7. 迷迭香石油醚提取物用于制备治疗或预防高脂血症、高血糖、非酒精性脂肪肝、糖尿病肾病、2型糖尿病中任意一种或多种的组合物。
  8. 根据权利要求7中的组合物,其特征为：
    用于制备治疗或预防I型或II型糖尿病的组合物；或；
    用于制备保护胰岛细胞、修复受损的胰岛细胞的组合物；或；
    用于制备保护糖尿病肾病人的肾脏组织、或、修复糖尿病肾病人的受损的肾脏组织的组合物。
  9. 根据权利要求7中的组合物,其特征为：迷迭香石油醚提取物的组分是：鼠尾草酸、鼠尾草酚、迷迭香酚、表迷迭香酚、表异迷迭香酚、鼠尾酸甲酯、Salviol、1-Phenanthrenecarboxylic acid、2-Phenanthrenemethanol。
  10. 根据权利要求7中的组合物,其特征为：迷迭香石油醚提取物的组分是：迷迭香酚，鼠尾草酚，鼠尾草酸，鼠尾酸甲酯，Salviol；其相对质谱峰面积比值为：2.23：10.12：39.64：2.93：32.32。
  11. 根据权利要求7中的组合物,其特征为：
    用于制备防治高脂血症、防治高血糖、或、非酒精性脂肪肝、糖尿病肾病、2型糖尿病的组合物，迷迭香石油醚提取物的含量范围是1毫克-2克。
  12. 根据权利要求7中的组合物,其特征为：
    迷迭香石油醚提取物的制备方法是：醇提取迷迭香药材，用水溶解醇提取物，用石油醚 萃取水溶液，合并石油醚层，回收石油醚得迷迭香石油醚部位。
  13. 根据权利要求7中的组合物,其特征为：
    迷迭香石油醚提取物的制备方法是：
    迷迭香药材用乙醇提取，合并95％乙醇提取液，回收乙醇，浸膏加水使溶，加入石油醚，石油醚和溶液萃取多次，合并石油醚层，回收石油醚得迷迭香石油醚部位；
    迷迭香药材用甲醇提取，合并甲醇提取液，回收甲醇，浸膏加水使溶，加入石油醚，石油醚和溶液按照萃取多次，合并石油醚层，回收石油醚得迷迭香石油醚部位；
    迷迭香药材用正丁醇提取，合并正丁醇提取液，回收正丁醇，浸膏加水使溶，加入石油醚，石油醚和溶液萃取多次，合并石油醚层，回收石油醚得迷迭香石油醚部位。
  14. 根据权利要求7中的组合物,其特征为：
    组合物制成：药物、保健品、或功能性食品,赋形剂或载体为制药或食品领域中常用的赋形剂或载体，如稀释剂，崩解剂，润滑剂等。
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CN112618586A (zh) * 2020-12-29 2021-04-09 温州医科大学 菜头肾总酚酸提取物的应用
CN112618586B (zh) * 2020-12-29 2021-12-28 温州医科大学 菜头肾总酚酸提取物的应用
CN118924723A (zh) * 2024-07-19 2024-11-12 中国中医科学院医学实验中心 鼠尾草酸在制备用于治疗或改善代谢相关脂肪性肝病的药物中的应用

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