CN107890466A - A kind of chromene lactone is used for the purposes that for aristolochic acid and the Chinese medicine containing aristolochic acid is attenuated - Google Patents
A kind of chromene lactone is used for the purposes that for aristolochic acid and the Chinese medicine containing aristolochic acid is attenuated Download PDFInfo
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- CN107890466A CN107890466A CN201711263828.6A CN201711263828A CN107890466A CN 107890466 A CN107890466 A CN 107890466A CN 201711263828 A CN201711263828 A CN 201711263828A CN 107890466 A CN107890466 A CN 107890466A
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- aristolochic acid
- chromene
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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
- A61K31/33—Heterocyclic compounds
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/365—Lactones
- A61K31/366—Lactones having six-membered rings, e.g. delta-lactones
- A61K31/37—Coumarins, e.g. psoralen
Abstract
The invention discloses a kind of chromene lactone to be used for the purposes that for aristolochic acid and the Chinese medicine containing aristolochic acid is attenuated.Chromene lactone provided by the invention can reduce the renal toxicity of aristolochic acid, can be used for preparing the single medicinal material containing aristolochic acid or Chinese medical extract or the toxicity-reducing medicament of compound Chinese medicinal preparation.Above-mentioned chromene lactone can both develop into single medicine, take, the single medicinal material containing aristolochic acid or Chinese medical extract or Chinese medicine compound prescription can also be developed into pharmaceutical composition with above-mentioned chromene lactone simultaneously with the single medicinal material containing aristolochic acid or Chinese medical extract or compound Chinese medicinal preparation.Chinese medicine can be the Chinese medicine containing aristolochic acid such as caulis aristologhiae manshuriensis, birthwort, Aristolochia fangchi, dutchmanspipe root, Ciliatenerve Knotweed Root, berba aristolochiae mollissimae, herba aristolochiae, the bark of official magnolia, asarum.Chinese medicine compound prescription can be the Chinese medicine compound prescriptions containing aristolochic acid such as rheum officinale clearing stomach ball, children-welfare tablet, Fenqing Wulin pill, Longdan Xiegan wan, shixiang fansheng pill, storax pill for treating coronary heart disease.
Description
Technical field
The invention belongs to field of medicaments, is related to chromene lactone and is used for aristolochic acid and the Chinese medicine attenuation containing aristolochic acid
Purposes.
Background technology
On October 18th, 2017, the researcher from Singapore and TaiWan, China is in Science periodicals Science
Translational Medicine issue entitled " Aristolochic acids and their derivatives are
Widely implicated in liver cancers in Taiwan and throughoutAsia " research paper.Should
Article points out dark aspect (toxic side effect) of Chinese herbal medicine, its leading role-aristolochic acid in the form of cover story
(aristolochic acids, AA) is a kind of chemistry being widely present in aristolochiaceae plant and a variety of natural herbals into
Point.The chemical constitution of four kinds of main aristolochic acids is as follows：
The toxicity that state food pharmaceuticals administration general bureau of China (CFDA) and the world of medicine also pay special attention to aristolochic acid is made
With.The China that is operated in that standardized administration contains Aristolochic Acid compound Chinese herbal medicine deploys already, and is continued for carrying out.
The case of acute renal failure has been caused to be reported excessive use Aristolochiaceae caulis aristologhiae manshuriensis early in Wu Han pines in 1964
(bibliography：Wu Songhan, two reports of akebi induced Acute renal failure, Jiangsu traditional Chinese medicine, 1964).90 years 20th century
Generation, the generation of the event such as " Belgian slimming drugs (Aristolochia fangchi) ", " Longdan Xiegan wan ", and Chinese patent drug containing aristolochic acid trigger
The case quantity of kidney function damage is also on the increase, and promotes the toxicity research of the Chinese herbal medicine containing aristolochic acid further to go deep into, manage
It is stricter.Version in 2000《Chinese Pharmacopoeia》Carried out under caulis aristologhiae manshuriensis kind using limiting, being indicated in the case where paying attention to item " can not be more
With, long term usage；Renal insufficiency and be not taken by pregnant women "；CFDA took again in the medicinal standard for eliminating caulis aristologhiae manshuriensis in 2003,2004
The medicinal standard of the Aristolochia fangchi that disappeared and dutchmanspipe root, and propose to strengthen the Chinese medicine containing birthwort, berba aristolochiae mollissimae, herba aristolochiae and Ciliatenerve Knotweed Root
The supervision and management of preparation；Version in 2005《Chinese Pharmacopoeia》Also start to phase out the Chinese herbal medicine containing aristolochic acid in record, extremely
Version in 2015《Chinese Pharmacopoeia》Only 3 kinds of herba aristolochiae, birthwort and asarum medicines containing aristolochic acid still record wherein.
But have much that the Chinese medicine containing aristolochic acid has its special purposes in tcm field, and do not have also at present
There is the side's of the entering substitute for searching out these Chinese medicines containing aristolochic acid.This results in some Gus for passing through succession in hundreds and thousands of years
Allusion quotation recipe cannot be further continued for using because of the renal toxicity of aristolochic acid, tangible sorry.
The content of the invention
It is aristolochic acid to be used for it is an object of the invention to overcome the deficiencies of the prior art and provide a kind of chromene lactone
And the purposes of the Chinese medicine attenuation containing aristolochic acid, for reducing the renal toxicity of the Chinese medicine containing aristolochic acid.
The above-mentioned purpose of the present invention is achieved by following technical scheme：
The chromene lactone of following chemical constitution is used for the purposes for preparing aristolochic acid toxicity-reducing medicament：
Preferably, the attenuation refers to the renal toxicity for reducing aristolochic acid.
Preferably, the aristolochic acid is Aristolochic acid-I, Aristolochic acid-I I, Aristolochic acid-I II and Aristolochic acid-I V
In one or more.
Above-mentioned chromene lactone is used for the purposes for preparing the toxicity-reducing medicament of the single medicinal material containing aristolochic acid；It is described to subtract
Poison refers to the renal toxicity for reducing the single medicinal material containing aristolochic acid.
Above-mentioned chromene lactone is used for the purposes for preparing the toxicity-reducing medicament of the Chinese medical extract containing aristolochic acid；It is described
Attenuation refers to the renal toxicity for reducing the Chinese medical extract containing aristolochic acid.
Preferably, the Chinese medicine is caulis aristologhiae manshuriensis, birthwort, Aristolochia fangchi, dutchmanspipe root, Ciliatenerve Knotweed Root, berba aristolochiae mollissimae, herba aristolochiae, thickness
Piao, asarum etc..
Above-mentioned chromene lactone is used for the purposes for preparing the toxicity-reducing medicament of the compound Chinese medicinal preparation containing aristolochic acid；Institute
State attenuation and refer to the renal toxicity for reducing the compound Chinese medicinal preparation containing aristolochic acid.
Preferably, the compound Chinese medicinal preparation be rheum officinale clearing stomach ball, children-welfare tablet, Fenqing Wulin pill, Longdan Xiegan wan,
Shixiang fansheng pill, storax pill for treating coronary heart disease etc..
A kind of pharmaceutical composition, including the single medicinal material containing aristolochic acid, Chinese medical extract or Chinese medicine compound prescription, in addition to
Above-mentioned chromene lactone.
Advantages of the present invention：
It is a discovery of the invention that chromene lactone provided by the invention can reduce the renal toxicity of aristolochic acid, can be used for
Prepare the single medicinal material containing aristolochic acid or Chinese medical extract or the toxicity-reducing medicament of compound Chinese medicinal preparation.Above-mentioned chromene
Lactone can both develop into single medicine, with the single medicinal material containing aristolochic acid or Chinese medical extract or compound Chinese medicinal preparation
Take simultaneously, also can be by the single medicinal material containing aristolochic acid or Chinese medical extract or Chinese medicine compound prescription and above-mentioned chromene lactone
Develop into pharmaceutical composition.
Brief description of the drawings
Fig. 1 is each group HK-2 Apoptosis ratio (%)；
Fig. 2 is that each group renal interstitial fibrosis rat relative area compares；
Fig. 3 is each group renal tissues of rats markers of fibrosis PROTEIN C OL- I relative expression levels.
Embodiment
Essentiality content of the present invention is specifically introduced with reference to the accompanying drawings and examples, but the guarantor of the present invention is not limited with this
Protect scope.
The chromene lactone that following embodiments use is the compound of following structure, and purchase or self-control, purity are not less than
95%.
Embodiment 1：The protective effect for causing people's renal cells to damage to aristolochic acid
First, experiment material
People's kidney proximal tubular cell line (HK-2) is purchased from China typical culture collection center；
Aristolochic acid-I (AA- I) is purchased from Nat'l Pharmaceutical ＆ Biological Products Control Institute；
Hyclone (FCS), Gibico companies；DMEM：HamsF-12 culture mediums (Hyclone)；Annexin-V reagents
Box, Invitrogen companies；
Enzyme-linked immunosorbent assay instrument, Thermofisher companies of the U.S.；Flow cytometer, U.S. company BD.
2nd, experimental method
1st, cell culture and packet
Well-grown HK-2 cells are taken, are suspended in the DMEM containing 10%FCS and 1% penicillin+streptomysin：HamsF-12
In culture medium, with 6 × 107/ L cell suspension inoculations are inoculated with 100 μ L per hole, in 37 DEG C, 5%CO in 96 orifice plates2, humidity
After cultivating 24h cell attachments under 100% environment, nutrient solution is abandoned in suction, continues to be incubated training by the following relative medicine that is separately added into
Support 48h.6 groups are set up in experiment altogether, and each group sets 8 multiple holes：
(1) control group：Nutrient solution is not added with AA- I；
(2) aristolochic acid group：AA- I is added in nutrient solution (25mg/L, each pharmaceutical intervention group are same)；
(3) netvein goldenray root element A groups：Netvein goldenray root element A (10mg/L, being final concentration, similarly hereinafter) and AA- I is added in nutrient solution simultaneously；
(4) netvein goldenray root element B groups：Netvein goldenray root element B (10mg/L) and AA- I is added in nutrient solution simultaneously；
(5) netvein goldenray root element C groups：Netvein goldenray root element C (10mg/L) and AA- I is added in nutrient solution simultaneously；
(6) netvein goldenray root element group：Add netvein goldenray root plain (10mg/L) and AA- I simultaneously in nutrient solution；
2nd, mtt assay measure cell survival rate
Each group 4h before culture terminates, the 5mg/L μ L of MTT solution 20 are added per hole, after culture terminates, 100 μ are added per hole
L DMSO, using the OD values that each hole is measured at enzyme-linked immunosorbent assay instrument 480nm.
3rd, flow cytometry detection apoptosis rate
After culture terminates, cell 1 × 10 is collected6Individual/mL, 1000rpm centrifuge 5min, abandon supernatant, and cell is resuspended with PBS,
Operated according to Annexin-VFITC cell apoptosis detection kits specification, detect the percentage of apoptotic cell and non-viable non-apoptotic cell.
4th, statistical method
Analyzed using SPSS19.0 software statistics, data are represented with mean value ± deviation, are compared between group using single factor test variance
Analysis.
3rd, experimental result
1st, mtt assay measure cell survival rate
Compared with control group, after AA- I acts on 48h, aristolochic acid group HK-2 cytoactives are obvious suppressed (P ＜ 0.05)；
Compared with aristolochic acid group, netvein goldenray root element A groups, netvein goldenray root element B groups, netvein goldenray root element C groups, netvein goldenray root element group HK-2 cytoactives
Significantly improve (P ＜ 0.05).OD values under the conditions of each group 480nm wavelength detectings are shown in Table 1.
OD values under the conditions of each group 480nm wavelength detectings of table 1
2nd, flow cytometry detection apoptosis rate
Compared with control group, after AA- I acts on 48h, aristolochic acid group HK-2 Apoptosis ratios substantially increase (P ＜
0.05)；Compared with aristolochic acid group, netvein goldenray root element A groups, netvein goldenray root element B groups, netvein goldenray root element C groups, netvein goldenray root element group HK-2 are thin
Born of the same parents' apoptosis ratio substantially reduces (P ＜ 0.05).Each group apoptosis value is shown in Table 2 and Fig. 1.
The each group HK-2 Apoptosis ratios of table 2
The embodiment illustrates that Aristolochic Acid in Antagonizing Human Renal Tubular epithelial cell HK-2 has obvious cytotoxicity, can caused
HK-2 survival rates reduce, apoptosis rate rise；Chromene lactone compound netvein goldenray root element A provided by the invention, netvein goldenray root element B,
Netvein goldenray root element C, netvein goldenray root element can have exploitation with antagonism Aristolochic Acid in Antagonizing Human Renal Tubular epithelial cell HK-2 cytotoxicity
Into the prospect of aristolochic acid toxicity-reducing medicament.
Embodiment 2：Cause the protective effect of renal interstitial fibrosis rat to aristolochic acid
First, experiment material
SPF level male SD rats, body weight 280-300g, it is purchased from Jiangning county Qinglongshan animal reproduction field；
3,4-methylenedioxy-8-methoxy-10-nitro-1-phenanthrenecarboxylic acid is dissolved with edible oil, as gavage working solution.
2nd, experimental method
1st, aristolochic acid kidney fibrosis rat model and experiment packet are established
Aristolochic acid kidney fibrosis modeling method (bibliography：The foundation of Chronic Aristolochic Acid Nephropathy animal model and ox
Sulfonic acid is acted on its early protection, the special collection of Chinese combination of Chinese tradiational and Western medicine magazine in June, 2003 fundamental research of volume 23, Beijing
Department of pathology of University Hospital portion)：Rat is first pressed to dosage the continuous gavage 5d, drug withdrawal 9d of 20mg/kg/d 3,4-methylenedioxy-8-methoxy-10-nitro-1-phenanthrenecarboxylic acids；Hereafter
Dosage reduces to 15mg/kg/d, every other week gastric infusion, until the 11st week.
After SPF level male SD rats adaptability is fed one week, 6 groups, every group 10 are randomly divided into according to body weight：
(1) control group：Gavage gives isometric solvent (edible oil)；
(2) aristolochic acid group：Modeling according to the method described above；
(3) netvein goldenray root element A groups：Modeling as stated above, and from first week the next day gavage 25mg/kg/d netvein goldenray root elements A；
(4) netvein goldenray root element B groups：Modeling as stated above, and from first week the next day gavage 25mg/kg/d netvein goldenray root elements B；
(5) netvein goldenray root element C groups：Modeling as stated above, and from first week the next day gavage 25mg/kg/d netvein goldenray root elements C；
(6) netvein goldenray root element group：Modeling as stated above, and from first week the next day gavage 25mg/kg/d netvein goldenray roots element；
2nd, renal fibrosis pathologic finding
After culture terminates, put to death rat, take out rat kidney, PBS elutes totally on ice, conventional dehydration, embed,
After section and Masson dyeing, the renal interstitial image of light microscope (× 100) collection cortical section (adopt at random by every rat
Collect 15 visuals field), carry out automatic measurement with multi-functional true color pathological image analysis system (BJ University of Aeronautics ＆ Astronautics manufactures)
Analysis, the kidney region fibrosis relative area=renal interstitial Lv Ran areas area/visual field gross area × 100%.
3rd, the detection that renal fibrosis marker protein COL- I is expressed
After culture terminates, rat is put to death, rat kidney is taken out, is positioned on ice, prevents protein degradation.Clip about 35mg
Tissue, the lysate that 350 μ L concentration are 100mg/mL is added, is fully ground with grinding rod, stands 20min on ice, phase at 4 DEG C
To centrifugal force 5.67 × 104× g centrifuges 15min.Protein supernatant is drawn, discards precipitation.RCF 5.67 at 4 DEG C
×104× g centrifuges 15min, draws protein supernatant again, discards precipitation.Using GAPDH albumen as internal reference, using protein
Blotting (westernblot) measure each group renal tissues of rats COL- I relative expression levels.
4th, statistical method
Analyzed using SPSS19.0 software statistics, data are represented with mean value ± deviation, are compared between group using single factor test variance
Analysis.
3rd, experimental result
1st, each group renal interstitial fibrosis rat relative area
Compared with control group, aristolochic acid group renal interstitial fibrosis rat relative area significantly raises (P ＜ 0.05)；With horse
Pocket bell acid group compares, netvein goldenray root element A groups, netvein goldenray root element B groups, netvein goldenray root element C groups, netvein goldenray root element group renal interstitial fibrosis rat
Relative area substantially reduces (P ＜ 0.05).Each group renal interstitial fibrosis rat relative area is shown in Table 3 and Fig. 2.
The renal interstitial fibrosis rat relative area (%) of table 3
2nd, the expressions of each group renal tissues of rats markers of fibrosis PROTEIN C OL- I
Compared with control group, the protein expression levels of COL- I significantly raise (P ＜ in aristolochic acid group renal tissues of rats
0.05)；Compared with aristolochic acid group, netvein goldenray root element A groups, netvein goldenray root element B groups, netvein goldenray root element C groups, netvein goldenray root element group Rat renal
The protein expression levels of COL- I substantially reduce (P ＜ 0.05) in tissue.Each group rat COL- I relative expression levels such as Fig. 3
It is shown.
The embodiment illustrates that aristolochic acid can cause renal tissues of rats fibrosis；Chromene lactone provided by the invention
Compound netvein goldenray root element A, netvein goldenray root element B, netvein goldenray root element C, netvein goldenray root element can have with renal toxicity inside antagonism aristolochic acid
There is prospect of the exploitation into aristolochic acid toxicity-reducing medicament.
Embodiment 3：Pharmaceutical composition is made with single medicinal material
A kind of pharmaceutical composition, it is in single medicinal material and netvein goldenray root element A, netvein goldenray root element B, netvein goldenray root element C, netvein goldenray root element
One or more combinations；The single medicinal material can be caulis aristologhiae manshuriensis, birthwort, Aristolochia fangchi, dutchmanspipe root, Ciliatenerve Knotweed Root, berba aristolochiae mollissimae,
The Chinese medicine containing aristolochic acid such as herba aristolochiae, the bark of official magnolia, asarum.
Embodiment 4：Pharmaceutical composition is made with Chinese medical extract
A kind of pharmaceutical composition, it is in Chinese medical extract and netvein goldenray root element A, netvein goldenray root element B, netvein goldenray root element C, netvein goldenray root element
One or more combinations；The Chinese medicine can be caulis aristologhiae manshuriensis, birthwort, Aristolochia fangchi, dutchmanspipe root, Ciliatenerve Knotweed Root, berba aristolochiae mollissimae, day
The Chinese medicine containing aristolochic acid such as celestial rattan, the bark of official magnolia, asarum.
The pharmaceutical composition can be used for pharmacy or the products such as slim tea be made into.
Embodiment 5：Pharmaceutical composition is made with Chinese medicine compound prescription
A kind of pharmaceutical composition, it is in Chinese medicine compound prescription and netvein goldenray root element A, netvein goldenray root element B, netvein goldenray root element C, netvein goldenray root element
One or more combinations；The Chinese medicine compound prescription can be rheum officinale clearing stomach ball, children-welfare tablet, Fenqing Wulin pill, Longdan Xiegan wan,
The Chinese medicine compound prescription containing aristolochic acid such as shixiang fansheng pill, storax pill for treating coronary heart disease.
To sum up, chromene lactone provided by the invention can reduce the renal toxicity of aristolochic acid, can be used for preparation and contain
There are the single medicinal material or Chinese medical extract or the toxicity-reducing medicament of compound Chinese medicinal preparation of aristolochic acid.Above-mentioned chromene lactone was both
Single medicine can be developed into, with the single medicinal material containing aristolochic acid or Chinese medical extract or compound Chinese medicinal preparation simultaneously
Take, can also be by the single medicinal material containing aristolochic acid or Chinese medical extract or Chinese medicine compound prescription and above-mentioned chromene lactone
Develop into pharmaceutical composition.
The effect of above-described embodiment is the specific essentiality content for introducing the present invention, but those skilled in the art should know
Road, protection scope of the present invention should not be confined to the specific embodiment.
Claims (9)
1. the chromene lactone of following chemical constitution is used for the purposes for preparing aristolochic acid toxicity-reducing medicament：
2. purposes according to claim 1, it is characterised in that：The attenuation refers to the renal toxicity for reducing aristolochic acid.
3. purposes according to claim 2, it is characterised in that：The aristolochic acid be Aristolochic acid-I, aristolochic acid-
One or more in II, Aristolochic acid-I II and Aristolochic acid-I V.
4. the chromene lactone described in claim 1 is used for the use for preparing the toxicity-reducing medicament of the single medicinal material containing aristolochic acid
On the way；The attenuation refers to the renal toxicity for reducing the single medicinal material containing aristolochic acid.
5. the chromene lactone described in claim 1 is used for the toxicity-reducing medicament for preparing the Chinese medical extract containing aristolochic acid
Purposes；The attenuation refers to the renal toxicity for reducing the Chinese medical extract containing aristolochic acid.
6. the purposes according to claim 4 or 5, it is characterised in that：The Chinese medicine is caulis aristologhiae manshuriensis, birthwort, Aristolochia fangchi, green grass or young crops
The banksia rose, Ciliatenerve Knotweed Root, berba aristolochiae mollissimae, herba aristolochiae, the bark of official magnolia, asarum etc..
7. the chromene lactone described in claim 1 is used for the toxicity-reducing medicament for preparing the compound Chinese medicinal preparation containing aristolochic acid
Purposes；The attenuation refers to the renal toxicity for reducing the compound Chinese medicinal preparation containing aristolochic acid.
8. purposes according to claim 7, it is characterised in that：The compound Chinese medicinal preparation is rheum officinale clearing stomach ball, little Er Jin
Red piece, Fenqing Wulin pill, Longdan Xiegan wan, shixiang fansheng pill, storax pill for treating coronary heart disease etc..
9. a kind of pharmaceutical composition, including the single medicinal material containing aristolochic acid, Chinese medical extract or Chinese medicine compound prescription, its feature exist
In：Also include the chromene lactone described in claim 1.
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| CN201711263828.6A CN107890466B (en) | 2017-12-05 | 2017-12-05 | A kind of chromene lactone is used for as aristolochic acid and the purposes of the attenuation of the Chinese medicine containing aristolochic acid |
| PCT/CN2018/084057 WO2019109576A1 (en) | 2017-12-05 | 2018-04-23 | Use of benzopyran compound |
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| CN107987089A (en) * | 2017-12-06 | 2018-05-04 | 赵腾骅 | A kind of extract, preparation method and medical usage rich in chromene lactone |
| WO2019109576A1 (en) * | 2017-12-05 | 2019-06-13 | 赵腾骅 | Use of benzopyran compound |
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| WO2019109576A1 (en) * | 2017-12-05 | 2019-06-13 | 赵腾骅 | Use of benzopyran compound |
| CN107987089A (en) * | 2017-12-06 | 2018-05-04 | 赵腾骅 | A kind of extract, preparation method and medical usage rich in chromene lactone |
| CN107987089B (en) * | 2017-12-06 | 2018-09-04 | 南京道尔医药科技有限公司 | A kind of extract, preparation method and medical usage rich in chromene lactone |
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