CN105287448A - Application of antrodia camphorata compound in preparation of drugs for treating kidney diseases - Google Patents
Application of antrodia camphorata compound in preparation of drugs for treating kidney diseases Download PDFInfo
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- CN105287448A CN105287448A CN201410256389.6A CN201410256389A CN105287448A CN 105287448 A CN105287448 A CN 105287448A CN 201410256389 A CN201410256389 A CN 201410256389A CN 105287448 A CN105287448 A CN 105287448A
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Abstract
The invention discloses application of an antrodia camphorata compound in preparation of drugs for treating kidney diseases. The compound is shown by a formula I as shown in the specification, and the kidney diseases can be glomerulosclerosis or post-streptococcal glomerulonephritis.
Description
Technical field
The present invention relates to a kind of medical application of compound of separated from Antrodia camphorate, particularly relate to the purposes of a kind of Antrodia Camphorata compound at the medicine of preparation treatment kidney disease.
Background technology
Antrodia Camphorata (Antrodiacamphorata), also known as Antrodia camphorata, Antrodia camphorata or red Antrodia camphorata etc., belong to the perennial mushroom fungus class of Aphyllophorales (Aphyllophorales), Polyporaceae (Polyporaceae), for Taiwan endemic species fungus, be only grown on the rotten heartwood inwall of hollow of Taiwan child care class seeds-Cinnamomum kanahirai hay tree (CinnamoumkanehiraiHay).Because Cinnamomum kanahirai hay tree distributed quantity is very rare, add artificial felling trees unlawfully, more shape is rare to make to parasitize the wild Antrodia Camphorata quantity that wherein can grow, and due to its sporophore growth quite slow, trophophase is also only between June to October, and therefore price is very expensive.In Taiwan folk custom medically, Antrodia Camphorata has removing toxic substances, alleviating diarrhoea symptom, antiinflammatory, treatment liver related disease and the function such as anticancer.Antrodia Camphorata as the gill fungus mushrooms of edible medicinal, there is the composition of many complexity, in known physiologically active ingredient, comprise: triterpenoid compound (triterpenoids), polysaccharides (polysaccharides, as candy gathers in β-D-Portugal), adenosine (adenosine), vitamin is (as vitamin B, niacin), protein (containing immunoglobulin), super oxygen disproportionation ferment (superoxidedismutase, SOD), trace element (as: calcium, phosphorus, germanium), nucleic acid, steroid and blood pressure stabilization material (as antodiaacid) etc., this a little physiologically active ingredient is considered to have antitumor, increase immunocompetence, antiallergic, Anti-bacterium, resisting hypertension, the multiple efficacies such as blood sugar lowering and cholesterol reducing.Simultaneously in recent years, the special component having started to find successively wherein may have effect of anti-inflammatory, particularly can be used for treatment kidney disease etc.
Generally common kidney disease, is often through and attacks kidney pompon and affect renal function.Kidney pompon disease comprises many diseases that several genes and environmental factors cause, but nothing more than being mainly divided into kidney pompon sclerosis and the large class of acute glomerulonephritis two.Kidney pompon sclerosis refers to the kidney pompon sclerosis in kidney.Generally refer to the blood capillary of kidney, kidney pompon and damage in order to the renal function unit from blood filtration urine.Urine protein (measuring maximum protein in urine) is one of signal of kidney pompon sclerosis.Kidney injury can affect kidney filtration function, and causes albumen by blood leakage in urine.But kidney pompon sclerosis is only one of many origin causes of formation of urine protein.And Kidney sections may be one judge whether patient suffers from the necessary basis for estimation of kidney pompon sclerosis or other kidney diseases.Kidney pompon sclerosis, can mean locality kidney pompon sclerosis (FSGS) and nodositas kidney pompon sclerosis especially.
Locality kidney pompon sclerosis (FSGS) judges with the damage characteristic of the sclerosis of local kidney pompon and podocyte podocytic process flattening (footprocesseffacemen).The research display of nearly 20 years, suffers from its late stage renal disease incidence rate of FSGS sufferer between 13 to 78%.Although FSGS source of disease and pathogenic factor are not yet clear, should be mainly result from the infringement of kidney pompon epithelial cell itself, and cause in kidney pompon complicated reaction to produce, and then cause kidney pompon to harden.
Glomerulonephritis between nodositas kidney pompon sclerosis or blood capillary, also diabetes nephropathy (diabetic nephropathy) or Ji-Wei Er Shi disease (Kimmelstiel-Wilsonsyndrome) can be referred to as, be a kind of Progressive symmetric erythrokeratodermia kidney also, it results from the blood capillary pathological changes of kidney pompon.The feature of this disease is nephropathy and the sclerosis of diffusivity kidney pompon; And result from long-time having diabetes, and the primary treatment regimen of various countries is for washing kidney.
Up to now, although steroid and immune formulation are be mainly used in treating the method suffering from constitutional FSGS, but effect is still wired in the treatment that this little therapy develops for injury of kidney, and various side effect can be produced because of it, therefore Therapeutic Method is only based on empirical law, and less according to pathology evidence.
Acute glomerulonephritis then means the inflammatory response of kidney membrane tissue, and wherein membrane tissue has filtering function, and can be separated garbage from blood and more than fluid.
Modal acute glomerulonephritis is immunoglobulin A type acute glomerulonephritis (IgAN), cause the cause of disease of this nephritis accelerated development still unpredictable, and clinically still cannot Prevention and Curation, therefore be regarded as one of principal element of the chronic renal failure that may cause nephropathy.Therefore, in the kidney of IgAN sufferer, even if renal failure can be caused in upper, clinically and on pathology the factor rank of other immunity, but the abnormal increases such as systemic T cell activation and lymph corpuscle/macrophage/neutrophils infiltration, are regarded as causing one of IgAN main process converting chronic renal failure to.Moreover oxidative pressure is also for mainly causing in sufferer and the factor that in animal model, IgAN increases and develops; Once had report to point out, in the mankind and experimental nephropathy of majority (comprising IgAN), active oxygen (ROS) can be directly diseases induced.
Although candy matter Corticosterone (glucocorticoidsteroid) has been widely used in treatment IgAN sufferer, but its maintenance renal function for IgAN and to slow down the effect of urine protein still unclear, and because of the long-time side effect using the potential uncontrollable Immunosuppression reaction caused, still cause the problem in medication.
Experiment many has at present learnt that Antrodia Camphorata extract has many medical functions, and its ingredient also successively analyzed go out, but whether there is in Antrodia Camphorata extract the compound that other effectively can treat kidney disease, be still problem one of urgently to be resolved hurrily.
Summary of the invention
Accordingly, the present invention is the compound extracted from Antrodia Camphorata mainly for two kinds, detects it and whether has the effect can treating kidney disease and anti-kidney inflammation.
An object of the present invention is to provide the purposes of a kind of Antrodia Camphorata compound at the medicine of preparation treatment kidney disease, and this compound represents with formula I:
Wherein, this kidney disease is kidney pompon sclerosis or acute glomerulonephritis.
Preferably, wherein this kidney pompon sclerosis is locality kidney pompon sclerosis (FSGS) or nodositas kidney pompon sclerosis.
Preferably, wherein this acute glomerulonephritis is immunoglobulin A type acute glomerulonephritis (IgAN).
In above-claimed cpd, LT1 all has antioxidative effect for nephritis cell, it can thus be appreciated that, above-mentioned Antrodia Camphorata compound can be utilized future in the medicine of preparation treatment kidney disease, can expectedly for kidney disease known at present, comprise kidney pompon sclerosis and acute glomerulonephritis, particularly locality kidney pompon sclerosis (FSGS), nodositas kidney pompon sclerosis, immunoglobulin A type acute glomerulonephritis (IgAN) etc. all have effect of anti-inflammatory.
Accompanying drawing explanation
Figure 1A and Figure 1B figure represents the result of the cell survival rate analysis (MTTcellviabilityassay) of 2 kinds of Antrodia Camphorata compounds respectively.
Fig. 2 A and Fig. 2 B represents that 2 kinds of Antrodia Camphorata compounds are for antioxidative measurement result respectively.
Fig. 3 A and Fig. 3 B represents that 2 kinds of Antrodia Camphorata compounds carry out the analysis result of anti-inflammatory reaction (MCP-1) respectively.
Detailed description of the invention
The present invention from the compound of Antrodia Camphorata mainly for two kinds of extractions, detects it and whether has the effect can treating kidney disease and anti-kidney inflammation.AntrocamolLT1, AntrocamolLT2 are the noval chemical compound that inventor finds, will provide above-mentioned extraction and Structural Identification data below.
The extraction of Antrodia Camphorata composition
Get Antrodia Camphorata (Antrodiacamphorata) mycelium, after sporophore or the mixture of the two (1.0kg) extract 2 times with the ethanol of 10 times amount, merge concentrated can obtaining slightly to take out thing and be about 230g (LT-E), slightly take out thing and carry out distribution extraction 3 times with dichloromethane/water (1:1), dichloromethane layer is divided into be about 102.6g (LT-E-D) and water layer is about 127.4g (LT-E-W), get dichloromethane layer 6.0g with silica gel column chromatography through normal hexane/dichloromethane (1:4), dichloromethane, the solvent of ethanol/methylene (5:95) is separated, be divided into ANCA-E-D-1, ANCA-E-D-2, ANCA-E-D-3, ANCA-E-D-4 etc. four layers.
According to the above results, ANCA-E-D-2 and ANCA-E-D-3 is carried out purification further, ANCA-E-D-3 layer can be obtained a noval chemical compound AntrocamolLT1 through preparative anti-phase column chromatography (C-18 reverse-phase chromatography tubing string) with 80%MeOH/H20 near 18.75 minutes and be about 150mg, and LT-E-D-2 layer can obtain another noval chemical compound AntrocamolLT2 through preparative anti-phase column chromatography (C-18 reverse-phase chromatography tubing string) with 80%MeOH/H20 near 25.10 minutes is about 170mg.
AntrocamolLT1 is colourless product liquid, and the molecular formula of this compound is C by analysis
24h
38o
5, molecular weight is 406, and complete Chinese illustrious name is called 4-hydroxyl-2,3-dimethoxy-6-methyl-5-(9-hydroxyl-3,7,11-trimethyl-2,6,10-12 carbon triolefin)-2-cyclonene, 4-hydroxy-5-[9-hydroxy-3,7,11-trimethyldodeca-2,6,10-trienyl]-2,3-dimethoxy-6-methyl-cyclohex-2-enone.
LT1 Structural Identification data:
1h-NMR (400MHz, CDCl
3): 1.12 (3H, d, J=7.2Hz), 1.61 (3H, s), 1.64 (3H, s), 1.66 (3H, s), 1.68 (3H, s), 1.72 (1H, m), 1.98 – 2.30 (8H), 2.51 (1H, dq, J=11.6,7.2Hz), 3.62 (3H, s), 4.02 (3H, s), 4.33 (1H, d, J=2.8Hz), 4.35 (1H, dt, J=9.2,4.0Hz), 5.09 (1H, d, J=8.4Hz), 5.14 (1H, t, J=7.2Hz), 5.15 (1H, t, J=7.2Hz);
13c-NMR (100MHz, CDCl
3): 12.17 (q), 15.95 (q), 16.19 (q), 18.13 (q), 25.72 (q), 25.93 (t), 26.78 (t), 39.41 (t), 39.98 (d), 43.29 (d), 47.94 (t), 58.81 (q), 60.48 (q), 65.35 (d), 67.24 (d), 121.64 (d), 127.64 (d), 128.42 (d), 132.03 (s), 134.99 (s), 135.97 (s), 137.42 (s), 160.82 (s), 197.15 (s).
AntrocamolLT2: be colourless product liquid, the molecular formula of this compound is C by analysis
26h
40o
6; Molecular weight is 448, complete Chinese illustrious name is called 4-acetyl carboxyl-2,3-dimethoxy-6-methyl-5-(9-hydroxyl-3,7,11-trimethyl-2,6,10-12 carbon triolefin)-2-cyclonene, 4-acetoxy-5-[9-hydroxy-3,7,11-trimethyldodeca-2,6,10-trienyl]-2,3-dimethoxy-6-methyl-cyclohex-2-enone.
LT2 Structural Identification data:
1h-NMR (400MHz, CDCl
3): 1.18 (3H, d, J=7.2Hz), 1.54 (3H, s), 1.64 (3H, s), 1.67 (3H, s), 1.69 (3H, s), 1.72 (1H, m), 1.80 – 2.40 (8H), 2.50 (1H, dq, J=11.6,7.2Hz), 3.65 (3H, s), 3.98 (3H, s), 4.36 (1H, m), 5.10 (1H, t, J=6.8Hz), 5.12 (1H, d, J=8.0Hz), 5.20 (1H, t, J=6.4Hz), 5.72 (1H, t, J=3.2Hz),
13c-NMR (100MHz, CDCl
3): 12.80 (q), 15.96 (q), 16.09 (q), 18.14 (q), 20.93 (q), 25.72 (q), 26.19 (t), 26.76 (t), 39.47 (t), 41.25 (d), 42.98 (d), 48.12 (t), 59.65 (q), 60.67 (q), 65.53 (d), 68.98 (d), 120.74 (d), 127.42 (d), 128.25 (d), 131.74 (s), 134.70 (s), 137.31 (s), 137.56 (s), 158.21 (s), 169.73 (s), 196.84 (s).
Cell culture processes
This be use utilize mouse kidney diaphragm cell (MouseMClineCRL-1927) to carry out subsequent experimental.This cell strain takes from AmericanTypeCultureCollection (Rockville, MD, USA), with Dulbecco'smodifiedEagle'smedium and the Ham'sF-12medium culture fluid of 3:1 ratio, and add 5%fetalbovineserum and 14mMHEPES and regularly cultivate.
Cell survival rate is analyzed
Above-mentioned cell strain is cultivated 24 hours respectively in suitable culture fluid.By the cell after hypertrophy with PBS cleaning once, and process cell with the trypsin-EDTA of 1 times, under 1,200rpm centrifugal 5 minutes subsequently, cell Shen Dian is abandoned supernatant.Add the new culture fluid of 10ml afterwards, slight wobble makes cell again suspend, then is placed in by cell in 96 hole micro-plate.During test, respectively at the Antrodia Camphorata compound adding 0.01 ~ 200 μ g/ml in each hole, in 37 DEG C, 5%CO
2lower cultivation 48 hours.Thereafter, in each hole, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide (MTT) (the Sigma – Aldrich of 5mg/ml is added under the environment of lucifuge, St.Louis, MO) react 2 hours.Under 570nm extinction wavelength, measure its light absorption value with ferment immunity analysis instrument again, use and calculate the survival rate of cell, and extrapolate it and grow half suppression ratio desired concn (i.e. IC50 value) all experimental datas and all represent with mean+/-standard error.Experimental data with match t test (paired-ttest) carry out statistical analysis.Be less than 0.05 with p value and be considered as having statistically difference.
For the cell survival rate analysis (MTTcellviabilityassay) of 2 kinds of Antrodia Camphorata compounds, result is consulted as Figure 1A and Figure 1B.Result shows, and AntrocamolLT1, AntrocamolLT2 are in the (≤0.048 μ g/ml selected at present) concentration all do not affect kidney diaphragm Growth of Cells.
The effect of Antrodia Camphorata compound with oxidation resistance/anti-inflammatory
Due to known at present, kidney pompon sclerosis or acute glomerulonephritis are caused by serious inflammatory response, the leukocyte cell assembled because of immunoreation can disengage a large amount of ROS and cause nephrocyte internal oxidition adverse circumstance to produce, and then kidney is damaged, therefore reduction or even the oxidative pressure eliminated in nephrocyte are one of method for the treatment of pyelonephritis, therefore the ROS reduced in born of the same parents is also one of important indicator for the treatment of above-mentioned kidney disease.
The mensuration of reactiveoxygenspecies (ROS) in cell
In cell, the mensuration of ROS is determined by the fluorescence intensity of the oxide (2 ', 7 '-dichlorofluorescein) of detection 2 ', 7 '-dichlorofluoresceindiacetate.By kidney diaphragm cell with after testing compound process, add 2 ', 7 '-dichlorofluoresceindiacetate (2 μMs) reaction 30 minutes, then add LPS at fixed time.Then carry out survey with excitation wavelength 485nm with the absorption spectrum reader (Bio-RadLaboratories, Inc) of radiation wavelength 530nm to read, namely represent that ROS content is higher when fluorescence is stronger.
Because active oxygen (ROS) has inseparable relation with nephritis, carry out the mensuration of ROS in above-mentioned cell at this, result as shown in Figure 2 A and 2 B.
Consult shown in Fig. 2 A, inhibited oxidation pressure (ROS) analysis is carried out respectively with the AntrocamolLT1 of 0.012 μ g/ml, 0.048 μ g/ml, when the results are shown in 5 minutes, 0.012 μ g/ml can inhibited oxidation pressure 3%, 0.048 μ g/ml can inhibited oxidation pressure 14%.
Consult shown in Fig. 2 B, carry out inhibited oxidation pressure (ROS) analysis with the AntrocamolLT2 of 0.012 μ g/ml, 0.048 μ g/ml respectively, current concentration unrestraint effect when the results are shown in 5 minutes.
Result arranges as shown in following table one:
Table one
Antrodia Camphorata compound is for the anti-inflammatory effect of nephritis cell
Known at present, MCP-1 plays key player impelling body tubular tissue gap inflammatory response, renal tubules atrophy and renal fibrosis etc., therefore detects this index protein, judges the anti-inflammatory effect of Antrodia Camphorata compound for kidney cell according to this.
ELISA is utilized to detect cover group (Biosciences, LosAngeles, CA, USA), detect the CCL2 matter of cell conditioned medium liquid according to operation instruction, and utilize ELISA reader (Bio-Tek) to carry out survey to read (absorbing wavelength is 450nm).* * represents p<0.005, and NS represents.
Consult shown in Fig. 3 A, carry out anti-inflammatory reaction (MCP-1) with the LT1 of 0.048 μ g/ml and 0.012 μ g/ml and analyze, result shows current concentration unrestraint effect.
Consult shown in Fig. 3 B, carry out anti-inflammatory reaction (MCP-1) with the LT2 of 0.048 μ g/ml and 0.012 μ g/ml and analyze, result shows current concentration unrestraint effect.
Result arranges as shown in following table two:
Table two
According to above-mentioned experimental result, the experimental result measured from ROS in cell, LT1 all has antioxidative effect for nephritis cell, indirectly susceptible of proof its there are the potentiality of medicine for the preparation for the treatment of kidney disease, can expectedly for kidney disease known at present, comprise kidney pompon sclerosis and acute glomerulonephritis, particularly locality kidney pompon sclerosis (FSGS), nodositas kidney pompon sclerosis, immunoglobulin A type acute glomerulonephritis (IgAN) etc. all have effect of anti-inflammatory.
In addition, though LT2 temporary acomia incumbent what for nephritis cell, there is the both effectiveness of antioxidation or anti-inflammatory, exclude in protection scope of the present invention, but more can confirm the preciousness extracting equally and there is from the LT1 compound of Antrodia Camphorata anti-inflammatory effect by contrast.
Value in the true tool industry of purposes of the medicine of Antrodia Camphorata compound provided by the present invention in preparation treatment kidney disease, above describing is only preferred embodiment explanation of the present invention, those skilled in the art can do other all improvement according to above-mentioned explanation, these changes still belong in the scope of the claims defined in spirit of the present invention and claims.
Claims (3)
1. the purposes of Antrodia Camphorata compound in the medicine of preparation treatment kidney disease, this compound represents with formula I:
Wherein, this kidney disease is kidney pompon sclerosis or acute glomerulonephritis.
2. purposes as claimed in claim 1, wherein, this kidney pompon sclerosis is locality kidney pompon sclerosis (FSGS) or nodositas kidney pompon sclerosis.
3. purposes as claimed in claim 1, wherein, this acute glomerulonephritis is immunoglobulin A type acute glomerulonephritis (IgAN).
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101357883A (en) * | 2007-07-30 | 2009-02-04 | 国鼎生物科技股份有限公司 | Antrodia camphoratea pimelie kelone compound for treating autoimmune disease and medicine composition |
| CN103458884A (en) * | 2011-01-21 | 2013-12-18 | 国鼎生物科技股份有限公司 | Methods and compositions for treating kidney disorders |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101357883A (en) * | 2007-07-30 | 2009-02-04 | 国鼎生物科技股份有限公司 | Antrodia camphoratea pimelie kelone compound for treating autoimmune disease and medicine composition |
| CN103458884A (en) * | 2011-01-21 | 2013-12-18 | 国鼎生物科技股份有限公司 | Methods and compositions for treating kidney disorders |
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Application publication date: 20160203 |