CN103304518A - Sesquiterpenoids and pharmaceutical composition thereof, and application of pharmaceutical composition in pharmacy - Google Patents
Sesquiterpenoids and pharmaceutical composition thereof, and application of pharmaceutical composition in pharmacy Download PDFInfo
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- CN103304518A CN103304518A CN2013102361527A CN201310236152A CN103304518A CN 103304518 A CN103304518 A CN 103304518A CN 2013102361527 A CN2013102361527 A CN 2013102361527A CN 201310236152 A CN201310236152 A CN 201310236152A CN 103304518 A CN103304518 A CN 103304518A
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Abstract
The invention provides a pharmaceutical composition with two novel sesquiterpenoids obtained by separation and purification from a traditional Chinese medicine for promoting blood circulation and dissipating stasis serving as active ingredients, a preparation method of the pharmaceutical composition, and application of the pharmaceutical composition in preparation of a medicine for treating diabetic nephropathy or chronic nephrosis. The two novel sesquiterpenoids have remarkable effects of inhibiting a high-glucose induced rat glomerular mesangial cell from secreting IL-6 (Interleukin-6), fibronectin (Fibronectin) and IV type collagen and are in obvious dose-effect and time-effect relationship which reflects the application prospection of such sesquiterpenoids in the preparation of the medicine for treating diabetic nephropathy and chronic nephrosis.
Description
Technical field:
The invention belongs to technical field of pharmaceuticals, relate to sesquiterpene derivative and the pharmaceutical composition take it as activeconstituents thereof, its preparation method, with and application in the medicine of preparation treatment of diabetic nephropathy or chronic nephropathy.
Background technology:
The method of the harm of diabetic nephropathy or chronic nephropathy, pathomechanism and drug research and principle, we are in published patent publication No.: clear and definite description is all arranged among the CN102153630A.Really, modern nervous mode of life and the change of diet style cause fat in the world wide, diabetes to increase, itself so that lead to complications such as diabetic nephropathy or chronic nephropathy.In view of the present medicine shortage in this disease field or limited, therefore constantly seek the medicine of intervening diabetic nephropathy or chronic nephropathy very necessary.
In recent years progress shows that the genesis of diabetic nephropathy and ephrosis is and cytokine such as IL-6, and the excessive secretion of MCP-1 etc. and extracellular matrix such as type Ⅳ collagen and FN is closely related.Therefore observe the cytokine of the kidney mesangial cell that medicine induces high sugar and the impact of collagen protein, become the common method of present research medicine for treating diabetic nephropathy.
Dried lacquer is the product behind the resin drying of lacquer tree (Toxicodendron verniciflum) secretion, have another name called the lacquer slag, from ancient times medicinal, successive dynasties have 25 with the dried lacquer ball of dried lacquer name according to incompletely statistics, dried lacquer also is one of important composition flavour of a drug of famous classical prescription Radix Et Rhizoma Rhei et Eupolyphaga Seu Steleophaga Pilulae, has the promoting blood circulation and breaking stagnation effect.Diabetic nephropathy belongs to microangiopathies, and we infer might search out the anti diabetes and kidney disease active substance from invigorating blood circulation (breaking) stasis of blood Chinese medicine, be used for the intervention of the chronic nephropathys such as diabetic nephropathy.In the prior art there are no compound of the present invention and have diabetic nephropathy or chronic nephropathy is worth the relevant report of the activity for the treatment of prospect.
Summary of the invention:
The object of the present invention is to provide to have the compound 1 and 2 that anti diabetes and kidney disease or chronic nephropathy are worth, the pharmaceutical composition take it as activeconstituents, its preparation method, with and application in the medicine of preparation treatment of diabetic nephropathy or chronic nephropathy.
Above-mentioned purpose of the present invention is to be achieved by following technical scheme:
The pharmaceutical composition for the treatment of diabetic nephropathy or chronic nephropathy contains the above-claimed cpd 1 for the treatment of significant quantity or 2 and pharmaceutically acceptable carrier.
The application of compound 1 or 2 in the medicine of preparation treatment diabetic nephropathy or chronic nephropathy.
Prepare the method for described compound 1 and 2, get dried lacquer, be ground into meal, flood 3 times with 80% ethanol room temperature, each solvent 20L, united extraction liquid decompression and solvent recovery gets medicinal extract, use equal-volume ethyl acetate extraction 3 times behind the medicinal extract water suspendible, each 5L, the acetic acid ethyl acetate extract decompression and solvent recovery gets extract, this extract is through silica gel 200-300 order silica gel column chromatography, through 99:1,98:2,97:3,96:4,95:5,94:6,93:7,92:8,90:10,85:15,80:20 chloroform-methanol system gradient elution, every kind of solvent gradient is 2 times of column volumes, collects to get 10 components according to every part of 500mL; Component 5 is through silica gel 100g column chromatography, and the sherwood oil of 10:1-1:1-acetone gradient elution gets compound 2; Component 7 is through MCI gel CHP20P column chromatography, and 30 –, 100% methanol-water wash-out gets 4 inferior components, and wherein component 7-1 gets compound 1 through Sephadex LH-20 methyl alcohol column chromatography.
The compounds of this invention can directly be used separately or Combination application, also can comprise that plant milk extract forms the form use of compound with other medicines, can use different pharmaceutical excipients, makes many kinds of solids preparation and liquid preparation.Pharmaceutical composition of the present invention is used with the form of per weight dose.But medicine oral administration of the present invention and two kinds of form administrations of injection.Usage quantity can be carried out one or many according to variations such as the type of age of route of administration, patient, body weight, the disease for the treatment of and severity and be used.
Description of drawings:
Figure 1A-1C represents that compound suppresses the short inflammatory factor of kidney mesangial cell and extracellular matrix determination experiment that high sugar is induced: adopt the ELISA test kit of USCK company to detect Rat Mesangial culture supernatant IL-6, the expression of Collagen IV and Fibronectin; Figure 1A be compound to the restraining effect of interleukin-6 (IL-6), * P＜0.05vs normal glucose;
#The high sugar of P＜0.05vs; Figure 1B is compound to the restraining effect .*P of fibronectin (fibronectin)＜0.05vs normal glucose;
#The high sugar of P＜0.05vs; Fig. 1 C is compound to the restraining effect .*P of IV Collagen Type VI＜0.05vs normal glucose;
#The high sugar of P＜0.05vs.
Fig. 2 A-2C represents that compound suppresses the short inflammatory factor of kidney mesangial cell that high sugar induces and the time-effect relationship of extracellular matrix; Fig. 2 A is that compound is to the restraining effect (time-effect relationship) of interleukin-6 (IL-6); Fig. 2 B is that compound is to the restraining effect (time-effect relationship) of fibronectin (fibronectin); Fig. 2 C is that compound is to the restraining effect (time-effect relationship) of IV Collagen Type VI.
Embodiment:
The below further specifies essentiality content of the present invention with embodiments of the invention, but does not limit the present invention with this.Essence according to the present invention all belongs to scope of the present invention to the simple modifications that the present invention carries out.
Embodiment 1:
Compound 1 and 2 separation and purification:
Dried lacquer 17kg is ground into meal, floods 3 times with 80% ethanol room temperature, each solvent 20L.United extraction liquid decompression and solvent recovery gets medicinal extract, uses equal-volume ethyl acetate extraction 3 times behind the medicinal extract water suspendible, each 5L.The acetic acid ethyl acetate extract decompression and solvent recovery gets the 220g extract, this extract through silica gel column chromatography (silica gel 200-300 order, 2.5kg), chloroform-methanol system gradient elution (99:1,98:2,97:3,96:4,95:5,94:6,93:7,92:8,90:10,85:15,80:20), every kind of solvent gradient is 2 times of column volumes, collects according to every part of 500mL) must 10 components.Component 5 (18.3g) is through silica gel column chromatography (silica gel 100g), and sherwood oil-acetone (10:1-1:1) gradient elution gets compound 2 (4.2mg); Component 7 (13.0g) is through the MCIgelCHP20P column chromatography, and methanol-water (30 – 100%) wash-out gets 4 inferior components, and wherein component 7-1 (1.1g) is through SephadexLH-20(methyl alcohol) column chromatography, get compound 1 (5.0mg).
Table1
1Hand
13CNMR?data?of?1and2(δin?ppm,JinHz).
a1H?NMR?at400MHz,
13C?NMR?at100MHz?in?CDCl
3,and?the?assignments?were?based?on?DEPT,2D?NMR?experiments.
b1H?NMR?at500MHz,
13C?NMR?at100MHz?in?CDCl
3,and?the?assignments?were?based?on?DEPT,2D?NMR?experiments
Toxicodenane?B(1):colorless?gums;[α]
15 D+7.69(c0.24,CHCl
3);IR(KBr)ν
max3423,2958,2924,2853,1727,1462,1377,1159,1065,1033,968cm
-1;ESIMS(negative):m/z507[2M-H]
-;HRESIMS(negative):m/z253.1802[M-H]
-(calcd?for?C
15H
25O
3,253.1803).
Toxicodenane?C(2):colorless?gums;[α]
15 D–4.9(c0.35,CHCl
3);IR(KBr)ν
max3426,2954,2925,2854,1640,1463,1378,1168,1093,1042,1033,881cm
-1;EIMS:m/z236[M]
+;HREIMS:m/z236.1778[M]
+(calcd?for?C
15H
24O
2,236.1776).
Embodiment 2:
Compound 1 or 2 among the embodiment 1, method injects water routinely, and the essence filter can be made into injection liquid after the embedding sterilization.
Embodiment 3:
Compound 1 or 2 among the embodiment 1 is dissolved in it in sterile water for injection, filters with aseptic funnel, and packing, aseptic sealing by fusing namely gets powder injection behind the frozen drying.
Embodiment 4:
Some amount embodiment 1 compound 1 or 2 is prepared into various dose tablet formulation (such as table 1) with the vehicle auxiliary material: several vehicle auxiliary materials are evenly mixed with bulk drug, add 1% sodium cellulose glycolate solution and make in right amount soft material, the granulation of sieving, wet grain oven dry and the whole grain that sieves, adding Magnesium Stearate and talcum powder mix rear compressing tablet and get final product.
Bulk drug and the accessory formula of compound 1 or 2 medicinal composition tablets among table 1 embodiment 1
Embodiment 5:
Among the embodiment 1 compound 1 or 2 routinely method be equipped with various pharmaceutical excipients and can be made into capsule:
Contain compound 1 among the embodiment 1 or 2 preparations as the drug regimen capsule preparations of effective constituent, as the adjunct ingredient for preparing the medicinal composition capsule, proportioning is made the capsule preparations that contains compound 1 or 2 composition 1-100mg in every capsules among the use embodiment 1 in the compound any according to a certain percentage as active constituents of medicine, several vehicle of use.
Embodiment 6:
The anti diabetes and kidney disease of the compounds of this invention and the pharmaceutical composition that forms with pharmaceutical excipient thereof or the pharmacological action of chronic nephropathy.
Compound 1 and 2 suppresses kidney mesangial cell strain inflammatory factor and extracellular matrix determination experiment (see figure 1):
Adopt the ELISA test kit of Uscn company to detect Rat Mesangial culture supernatant IL-6, Collagen IV and Fibronectin express.Figure 1A is the compound dosage dependent expression of IL-6; Figure 1B is the compound dosage dependent expression of Fibronectin; Fig. 1 C is the compound dosage dependent expression of Collagen IV.Fig. 2 A is the compound time-effect relationship of IL-6; Fig. 2 B is the compound time-effect relationship of Fibronectin; Fig. 2 C is the compound time-effect relationship of Collagen IV.
Experimental principle:
Adopt double antibody sandwich method to measure Rat Mesangial culture supernatant IL-6, Fibronectin and Collagen IV expression level in the sample.Anti-IL-6, Fibronectin and Collagen IV antibody sandwich microwell plate with purifying, make insolubilized antibody, in the micropore of coated monoclonal antibody, add successively the testing sample that contains expression IL-6, Fibronectin and Collagen IV, again with the antibodies of HRP mark, form antibody-antigen-hrp-antibody complex, through thoroughly adding substrate TMB colour developing after the washing.TMB changes into blueness under the catalysis of HRP enzyme, and changes into final yellow under the effect of acid.The expression of the depth of color and the IL-6 in the sample, Fibronectin and Collagen IV is proportionate.With measuring absorbancy (OD value) under the microplate reader 450nm wavelength, by rat IL-6, Fibronectin in the typical curve calculation sample and Collagen IV concentration.
Sample process:
1) cells and supernatant: sterile tube is collected, 2,000rpm/min, centrifugal 20min.Carefully collect supernatant.Packing is frozen in-80 ℃.
2) scraping diapire cell, the cracking of total protein lysate, 12,000rpm/min, 4 ℃ of centrifugal 10min collect supernatant, and the Bradford method is measured total protein content.
ELISA detects:
Operation is undertaken by the test kit explanation:
1) dilution of standard substance and application of sample: the coated plate of enzyme mark is established the standard substance hole, and doubling dilution (each hole application of sample amount all is 50 μ l after the dilution) adds the standard substance of IL-6, Fibronectin and Collagen IV.
2) application of sample: establish respectively blank well (the blank hole does not add sample and enzyme marking reagent, and all the other each rapid operations are identical), testing sample hole (the diluted sample degree is 5 times).Rock gently mixing.
3) incubation: 37 ℃, 30min.
4) washing: discard liquid in the plate, dry, washing lotion is washed 5 times, 30 seconds/time.Thieving paper pats dry.
5) enzyme-added: every hole adds 50 μ l enzyme labelling liquid, except the blank well.
6) incubation: 37 ℃, 30min.
7) washing: discard liquid in the plate, dry, washing lotion is washed 5 times, 30 seconds/time.Thieving paper pats dry.
8) colour developing: every hole adds substrate A, each 50 μ l/ hole of B liquid.37℃，15min。Lucifuge.
9) stop: every hole adds each 50 μ l/ hole of stop buffer.
10) measure: with the blank well zeroing, microplate reader 450nm reads absorbancy (OD value).
11) experiment repeats 3 times.
The result calculates: utilize standard substance quantitative tracing analysis calculation sample concentration and with total protein of cell normalization (IL-6:pg/mg cell protein; Fibronectin﹠amp; Collagen IV:ng/mg cell protein).
Above presentation of results compound 1 and 2 is 1,3,10 and the kidney mesangial cell strain inflammatory factor IL-6, the Fibronectin that high sugar are induced during 30 μ M and Collagen IV restraining effect that dose-dependently is all arranged.Because these cytokines and extracellular matrix are detected in diabetic nephropathy and chronic nephropathy and present high expression level, the secretion that therefore suppresses them has positive value to the control of diabetic nephropathy or ephrosis undoubtedly.
Claims (4)
1. the sesquiterpenoids 1 and 2 that has following structural formula,
2. the pharmaceutical composition for the treatment of diabetic nephropathy or chronic nephropathy contains the compound claimed in claim 11 for the treatment of significant quantity or 2 and pharmaceutically acceptable carrier.
3. the application of compound claimed in claim 1 in the medicine of preparation treatment diabetic nephropathy or chronic nephropathy.
4. prepare the method for compound 1 claimed in claim 1 and 2, get dried lacquer, be ground into meal, flood 3 times with 80% ethanol room temperature, each solvent 20L, united extraction liquid decompression and solvent recovery gets medicinal extract, use equal-volume ethyl acetate extraction 3 times behind the medicinal extract water suspendible, each 5L, the acetic acid ethyl acetate extract decompression and solvent recovery gets extract, this extract is through silica gel 200-300 order silica gel column chromatography, through 99:1,98:2,97:3,96:4,95:5,94:6,93:7,92:8,90:10,85:15,80:20 chloroform-methanol system gradient elution, every kind of solvent gradient is 2 times of column volumes, collects to get 10 components according to every part of 500mL; Component 5 is through silica gel 100g column chromatography, and the sherwood oil of 10:1-1:1-acetone gradient elution gets compound 2; Component 7 is through MCI gel CHP20P column chromatography, and 30 –, 100% methanol-water wash-out gets 4 inferior components, and wherein component 7-1 gets compound 1 through Sephadex LH-20 methyl alcohol column chromatography.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103819437A (en) * | 2014-03-10 | 2014-05-28 | 中国科学院昆明植物研究所 | Spirolingzhine compounds as well as pharmaceutical composition and applications thereof |
| CN104447783A (en) * | 2014-11-07 | 2015-03-25 | 中国科学院昆明植物研究所 | Ganoderma cochlear phenols A and B, pharmaceutical compositions of ganoderma cochlear phenols A and B and applications of ganoderma cochlear phenols A and B and pharmaceutical compositions in preparation of medicines and food |
| CN106957324A (en) * | 2016-01-11 | 2017-07-18 | 复旦大学 | Sequiterpene spiro lactone compounds and its production and use |
| CN111153911A (en) * | 2019-10-12 | 2020-05-15 | 深圳大学 | Lipid-lowering compound comiphorine A and application thereof in pharmacy |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2009015014A2 (en) * | 2007-07-20 | 2009-01-29 | Shrier David L | Multi-step method of pain and/or inflammation treatment |
| EP2070544A2 (en) * | 2007-12-13 | 2009-06-17 | Marianna Gross | Means of withdrawal from smoking |
| WO2010057295A1 (en) * | 2008-11-18 | 2010-05-27 | Origin Biomed Inc. | Homeopathic composition comprising hypericum perforatum extract and essential oils for the treatment of neuropathic pain |
| CN101906122A (en) * | 2010-08-16 | 2010-12-08 | 江西山香药业有限公司 | Method for extracting rhoifolin and prepared medicament application thereof |
| CN102153630A (en) * | 2011-01-11 | 2011-08-17 | 中国科学院昆明植物研究所 | Ring octapeptide and preparation method and application thereof in medicament making |
-
2013
- 2013-06-14 CN CN201310236152.7A patent/CN103304518B/en not_active Expired - Fee Related
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2009015014A2 (en) * | 2007-07-20 | 2009-01-29 | Shrier David L | Multi-step method of pain and/or inflammation treatment |
| EP2070544A2 (en) * | 2007-12-13 | 2009-06-17 | Marianna Gross | Means of withdrawal from smoking |
| WO2010057295A1 (en) * | 2008-11-18 | 2010-05-27 | Origin Biomed Inc. | Homeopathic composition comprising hypericum perforatum extract and essential oils for the treatment of neuropathic pain |
| CN101906122A (en) * | 2010-08-16 | 2010-12-08 | 江西山香药业有限公司 | Method for extracting rhoifolin and prepared medicament application thereof |
| CN102153630A (en) * | 2011-01-11 | 2011-08-17 | 中国科学院昆明植物研究所 | Ring octapeptide and preparation method and application thereof in medicament making |
Non-Patent Citations (1)
| Title |
|---|
| JIANG-BO HE ET AL.: "Sesquiterpenoids with New Carbon Skeletons from the Resin of Toxicodendron vernicifluum as New Types of Extracellular Matrix Inhibitors", 《ORG. LETT.》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103819437A (en) * | 2014-03-10 | 2014-05-28 | 中国科学院昆明植物研究所 | Spirolingzhine compounds as well as pharmaceutical composition and applications thereof |
| CN104447783A (en) * | 2014-11-07 | 2015-03-25 | 中国科学院昆明植物研究所 | Ganoderma cochlear phenols A and B, pharmaceutical compositions of ganoderma cochlear phenols A and B and applications of ganoderma cochlear phenols A and B and pharmaceutical compositions in preparation of medicines and food |
| CN106957324A (en) * | 2016-01-11 | 2017-07-18 | 复旦大学 | Sequiterpene spiro lactone compounds and its production and use |
| CN106957324B (en) * | 2016-01-11 | 2019-02-26 | 复旦大学 | Sequiterpene spiro lactone compounds and its preparation method and application |
| CN111153911A (en) * | 2019-10-12 | 2020-05-15 | 深圳大学 | Lipid-lowering compound comiphorine A and application thereof in pharmacy |
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| Publication number | Publication date |
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| CN103304518B (en) | 2014-12-31 |
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