CN1150951A - Five compounds of Qingyang ginseng glucoside and its preparation method and application - Google Patents
Five compounds of Qingyang ginseng glucoside and its preparation method and application Download PDFInfo
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Abstract
A process for preparing 5 compounds, Cynanchi otophylli glucosides C,D,E,F and G and a process for application of them in pharmaceutical industry are disclosed. Said compounds can especially be used to prepare the medicines for hepatitides and Meniere's disease with advantages of wide available raw materials, high curative effect, stability and biologic utilization rate, and low poison.
Description
The present invention relates to Cynanchum otophvllum glucoside third, glucoside fourth, glucoside penta, glucoside the sixth of the twelve Earthly Branches, glucoside five compounds in heptan, and preparation method thereof and in pharmacy field especially in the preparation Antihepatitis medicament and the application in the anti-Meniere's medicine.
The general formula of five compounds that the present invention relates to is:
Compound 1 (Cynanchum otophvllum glucoside third)
Compound 2 (Cynanchum otophvllum glucoside fourth)
Compound 3 (Cynanchum otophvllum glucoside penta)
Compound 4 (Cynanchum otophvllum glucoside the sixth of the twelve Earthly Branches)
Compound 5 (Cynanchum otophvllum glucoside heptan)
R2＝CH
3CO
-
Five compounds of the tool said structure formula that extraction separation obtains from Asclepiadaceae Cynanchum plant Cynanchum otophvllum all do not have bibliographical information.
The object of the present invention is to provide five kinds of new compound Cynanchum otophvllum glucosides third that from the Cynanchum otophvllum plant, obtain, glucoside fourth, glucoside penta, glucoside the sixth of the twelve Earthly Branches, glucoside heptan, their preparation method, and the particularly application in anti-hepatitis of preparation and anti-Meniere's medicine in pharmacy field.
The method for preparing general formula (I) compound provided by the present invention is to adopt vegetable chemistry method extraction separation from the plant Cynanchum otophvllum.
Concrete, above-mentioned preparation method gets Asclepiadaceae Cynanchum plant Cynanchum otophvllum (Cynanchumotophyllum Schneid) root, extract 2-3 time 60-70 ℃ of heat with chloroform, reclaim dope behind the solvent with the sherwood oil degreasing that refluxes, getting substrate is the thick glucoside block in drug effect position, uses CCL4-AcoEt (95: 5; 85: 15v/v) extract successively (95: 5v/v) extract and (85: 15v/v) extract, the latter is through silica gel H medium pressure column chromatography and Rp-18 medium pressure column chromatography repeatedly, more respectively with chloroform+methyl alcohol (98: 2 and 95: 5v/v), (70: 30 and 80: 20v/v) the mixed solution wash-out got final product methyl alcohol+water.
Another kind of preparation method of the present invention gets Asclepiadaceae suede Calamus plant Cynanchum otophvllum (Cynanchumotophyllum Schneid) root, extract 2-3 time 60-70 ℃ of heat with ethyl acetate, reclaim dope behind the solvent with the sherwood oil degreasing that refluxes, getting substrate is the thick glucoside block in drug effect position, uses CCL4-AcoEt (95: 5; 85: 15v/v) extract successively (95: 5v/v) extract and (85: 15v/v) extract, the latter is through silica gel H medium pressure column chromatography and Rp-18 medium pressure column chromatography repeatedly, more respectively with chloroform+methyl alcohol (98: 2 and 95: 5v/v), (70: 30 and 80: 20v/v) the mixed solution wash-out got final product methyl alcohol+water.
The present invention provides the application of general formula (I) compound in the preparation Antihepatitis medicament simultaneously.
Above-mentioned application, the especially application in the medicine of preparation treatment chronic persistent hepatitis and chronic hepatitis disease.
The present invention provides the application of general formula (I) compound in preparation anti-Meniere's medicine in addition.
Hepatitis is the difficult and complicated illness of harm humans life and health, at present both at home and abroad the development Antihepatitis medicament is mainly from two class medicines---and developing antiviral, immunopotentiating agent or the immunomodulator, is being ucleosides and Sulfate of polysaccharide class at the antiviral drug of clinical study just in the recent period; Immunopotentiating agent is in disturbing plain and the vegetable polysaccharides body, and it is more sure that for example deoxidation Zovirax and alpha-interferon share the clinical effectiveness for the treatment of hepatitis B.Other medicines still do not have breakthrough progress.But said medicine mostly has bigger toxic side effect, nucleotide drug, particularly some quite serious toxic side effect appear in the alpha-interferon clinical application, according to Giustina etc. at document Giustina et al:Ital J.Gastroenterol, 26:203, report in 1994 is used alpha-interferon treatment chronic viral hepatitis and spirit depressing occurred, side effects such as autoimmune disorder, diabetes, cardiovascular disorder, hemolytic anemia, minority is died from various organ failures.Therefore develop the curative effect height, treatment hepatitis medicament that toxic side effect is little has become urgent problem.And Meniere is a kind of common frequently-occurring disease, and morbidity is 0.5% of a total population, and the whole world has 2500-3000 ten thousand beautiful patients Ni Ershi, middle aged women number in the majority approximately.This disease is a kind of paroxysmal disease, and its symptom comprises that paroxysmal dizzy, nausea and vomiting, eyeball are stayed looks or tremble, some also have fluctuation deafness, ear toot and the ear sensation of fullness, a few patients (about 20% patient) stays disable vertigo and forfeiture hearing.This disease is not because of still having at present definite pathological study, the effective medicine of therefore not suiting the medicine to the illness.U.S. nile disease claims Mei Nier syndrome (Menier ' syndrome), be the imprecise difficult and complicated illness of not having the harm humans health of active drug again of a kind of cause of disease again.The world selects some kinds of medicines to carry out exploratory treatment clinically from scopolamine, dimenhydrinate, Anisodamine, promethazine hydrochloride, coromegine, betahistine hydrochloride (betahistine), meclizine medicines such as (mecligine) at present, and effect can not be satisfactory.And said medicine mostly has bigger toxic side effect, and long-term pharmacological agent has irreversible toxic side effect, as deaf and regular ear toot etc.Therefore develop the little treatment Meniere's medicine of curative effect height, toxic side effect and become urgent problem the sixth of the twelve Earthly Branches.The present invention is based on this, from the Cynanchum otophvllum plant extraction separation have anti-hepatitis and the active new drug composition of anti-Meniere's disease, with five kinds of compounds shown in the general formula (I).
Compound 1 Cynanchum otophvllum glucoside third molecular weight of the present invention is 630, white powder, and fusing point is 159-162 ℃, is dissolved in ethanol or other organic solvent.
Compound 2 Cynanchum otophvllum glucoside fourth molecular weight of the present invention are 1056, white powder, and fusing point is 154-157 ℃, is dissolved in ethanol or other organic solvent.
Compound 3 Cynanchum otophvllum glucosides penta molecular formula of the present invention is C48H76016, and molecular weight is 908, and white powder is dissolved in ethanol or other organic solvent.
Compound 4 Cynanchum otophvllum glucoside molecular formula in the sixth of the twelve Earthly Branches of the present invention are C48H76O16, and molecular weight is 908, and white powder is dissolved in ethanol or other organic solvent.
Compound 5 Cynanchum otophvllum glucoside molecular formula in heptan of the present invention are C44H70O16, and molecular weight is 854, and white powder is dissolved in ethanol or other organic solvent.
In order to understand essence of the present invention better, immune pharmacology, the toxicity with Cynanchum otophvllum glucoside third, glucoside fourth, glucoside penta, glucoside the sixth of the twelve Earthly Branches, glucoside compound in heptan illustrates its new purposes and beneficial effect thereof in the medicine of anti-hepatitis disease of preparation and anti-Meniere's disease below.
Adopt Cynanchum otophvllum glucoside third that embodiments of the invention make, glucoside fourth, glucoside penta, glucoside the sixth of the twelve Earthly Branches, glucoside to be used for following experiment heptan:
One, acute toxicity test:
50 of rats, be divided into 5 groups at random, every group 10, irritate the starch suspension that stomach gives the Cynanchum otophvllum glucoside third of various dose or glucoside fourth or glucoside penta or glucoside the sixth of the twelve Earthly Branches or glucoside heptan, observe toxicity symptom and all mortality ratio, ask medium lethal dose (LD50) by simplifying probit method, Cynanchum otophvllum glucoside third is 115mg/kg, and Cynanchum otophvllum glucoside fourth is 110mg/kg, and Cynanchum otophvllum glucoside penta is 120mg/kg, Cynanchum otophvllum glucoside the sixth of the twelve Earthly Branches is 118mg/kg, and Cynanchum otophvllum glucoside heptan is 124mg/kg.
The acute poisoning symptom is mainly excitement and dyskinesia, and recurrent tonic convulsion is a feature.
Two, sub-acute toxicity test:
Get 15-18 and restrain mouse 120 grams that then wean, male and female half and half, by body weight, sex random packet, every group 30, first, second and third group is irritated stomach respectively and is given about 1/10,1/20 and the starch suspension in the Cynanchum otophvllum glucoside third of 1/30LD50 or glucoside fourth or glucoside penta or glucoside the sixth of the twelve Earthly Branches or glucoside heptan, and control group is given regularly feeding of starch slurry, continuous 60 days, experimental session is weighed once every day, gets 1/3 animal when experiment finishes and does routine blood test and renal function (NPN) inspection.1/3 animal is cooked liver function (SGPT) and checks, 1/3 animal is observed after giving over to drug withdrawal, when observing blood picture and hepatic and renal function, carries out dirty and testis is weighed and obtained the organ pipe coefficient, and to the visual inspections such as gland, brain and testis of the heart, liver, spleen, lung, kidney, kidney.
In bimestrial subacute toxicity test, Cynanchum otophvllum glucoside third or glucoside fourth or glucoside penta or glucoside glucoside in the sixth of the twelve Earthly Branches dosage in heptan integral dose reach 5-6 times of LD50, most growth of animal are normal, show the oral no tangible cumulative effect of this medicine, but the big white mouse abdominal injection observed cumulative effect; Animal body body weight, red, total white blood cells, renal function and internal organs all there is not tangible detrimentally affect; Also observe in the experiment, Cynanchum otophvllum can change the ratio of lymphocyte and neutrophil, lymph is reduced and the neutrophil rising, but total white blood cells is not had obvious influence.
Three, the immune pharmacological action of anti-hepatitis:
With animal (mouse body weight 18-22 gram) random packet, every group 10, Cynanchum otophvllum glucoside third or glucoside fourth or glucoside penta or glucoside the sixth of the twelve Earthly Branches or glucoside add endoxan group (being called for short the Cynanchum otophvllum group) heptan, solvent polyoxyethylene glycol (PEG) adds endoxan group (being called for short the CP group), and blank group injecting normal saline every day 0.2ml/ only.The Cynanchum otophvllum group is pressed 1/4LD50 (70mg/kg) intramuscular injection every day, endoxan was pressed the 100mg/kg subcutaneous injection once on 10th, in experiment the 1st, 14,17,24,31 by tail vein blood mensuration total white blood cells, lymphocyte per-cent, ANAE positive lymphocyte per-cent, lymphocyte DNA content (pressing point-score):
1, to the influence of ANAE positive lymphocyte:
After giving CP the 4th day, CP group ANAE positive lymphocyte per-cent changed little, but its absolute number obviously descends, and dropped to minimum level on 7th behind injection CP, recovered normal then gradually; The decline in the 4th day behind injection CP of Cynanchum otophvllum group ANAE positive lymphocyte is not remarkable, drops to lower-most point on 7th yet, is evident as light but the decline degree is single with the CP group.
2, to the provide protection of lymphocyte DNA:
The content of CP group lymphocyte DNA began on 4th to descend, and dropped to minimum level on the 7th.The Cynanchum otophvllum group also is to begin in 4th to descend, and drops to lower-most point by 7 days, but all is significantly higher than the CP group.The dynamic change of lymphocyte DAN, the decline of the DNA value of CP group is rapid, and continues for some time; Though the dna content of Cynanchum otophvllum group has decline, the time generation that sharply descends is later, keeps the time weak point of the lower-most point of decline, with regard to bottom out.A large amount of cavitys all appear in CP group and PEG group lymphocyte nuclear dna colour attaching area, and the Cynanchum otophvllum group is not observed this phenomenon, and visible Cynanchum otophvllum has provide protection to lymphocyte DNA.
3, to the hepatitis patient Immune Effects:
Cynanchum otophvllum glucoside third or glucoside fourth or glucoside penta or glucoside the sixth of the twelve Earthly Branches or glucoside are treated delay property chronic hepatitis 44 examples respectively heptan, wherein extract 19 examples out and done the PHA tuerculoderma before and after treatment, and the mean diameter of blush is in the positive reaction of the above person of 14mm.The result is: having 2 examples to be the skin test feminine gender before the treatment, is 9.5mm before the 1 example treatment, rises to 16.5mm after the treatment, reaches clinical cure.Before the 1 example treatment is 5mm, rises to mean diameter 10.5mm after the treatment.(mean diameter is 14.5,16.0,15mm), rising is arranged all after the treatment, and its mean diameter is respectively 21.5mm, 17.5mm, 24mm to treat the positive weak reaction of preceding 3 examples.All at the positive reaction high level, there are 5 routine mean diameters that the 1-4mm of dwindling is arranged slightly after the treatment before other 14 example treatment, 2 routine no changes, 7 routine mean diameters increase from 5.5-16.5mm.19 examples before and after treatment the PHA mean diameter relatively, there were significant differences (t=3.1855，d.f＝19-1＝18，t18.005＝2.101p＜0.05)。Above result shows that Cynanchum otophvllum can make the PHA skin test reaction of hepatitis transfer the positive to, and high reactor is decreased, and prompting has regulating effect to the T lymphocyte.
Four, anti-Meniere's disease immunity pharmacological action
1, anti-audiogenic seizure
P π-PMc the rat of audiogenic seizure sensitivity is used in experiment, male and female are all used, random packet, every group 10, body weight 200-270 gram is at first measured outbreak susceptibility, continuous 3 days, react constant and in experiment, the ip this product of then dividing into groups is surveyed its anticonvulsion effect again after 4 hours, to prevent that fully outbreak from being index, calculate its ED50, the result proves that the ED50 of glucoside third is 10.8mg/kg, and the ED50 of glucoside fourth is 11.2mg/kg, the ED50 of glucoside penta is 10.5mg/kg, the ED50 in glucoside the sixth of the twelve Earthly Branches is 12.2mg/kg, and the ED50 in glucoside heptan is 10.8mg/kgg, and consumption has made outbreak obviously alleviate (P＜0.01) when 6.0-7.0mg/kg.
Take a broad view of above experimental result, beneficial effect of the present invention is:
Cynanchum otophvllum glucoside third of the present invention, glucoside fourth, glucoside penta, glucoside the sixth of the twelve Earthly Branches, glucoside tool in heptan medicine source is wide, drug effect is high, toxicity is low, good stability, advantage that the bio-absorbable availability is high:
Cynanchum otophvllum glucoside third of the present invention, glucoside fourth, glucoside penta, glucoside the sixth of the twelve Earthly Branches, glucoside have immunocompetence heptan, and toxic side effect is little, and convenient oral is the antihepatitis drug composition of effective low toxicity.Adopt Cynanchum plant Cynanchum otophvllum (Cynanchum otophyllum Schneid) root, extract 2-3 time 60-70 ℃ of heat with ethyl acetate, reclaim dope behind the solvent with the sherwood oil degreasing that refluxes, substrate be the thick glucoside block in drug effect position, with CCL4-AcoEt (95: 5; 85: 15v/v) extract successively (95: 5v/v) extract and (85: 15v/v) extract, the latter is through silica gel H medium pressure column chromatography and Rp-18 medium pressure column chromatography repeatedly, again respectively with chloroform+methyl alcohol, methyl alcohol+water mixed liquid wash-out gets Cynanchum otophvllum glucoside third respectively, the glucoside fourth, glucoside penta, glucoside the sixth of the twelve Earthly Branches, glucoside heptan, they are different from the single immunocompetence of general immune drug, they work simultaneously to immunity system-nervus centralis-internal secretion, this shows on pharmacology and the clinical effectiveness, collection is suffered from chronic hepatitis all over the body, the patient of impotence and epilepsy takes Cynanchum otophvllum glucoside third or glucoside fourth or glucoside penta or glucoside the sixth of the twelve Earthly Branches or glucoside and obtains the curative effect that a medicine is controlled three diseases heptan.Show that this medicinal ingredients oral absorption is good, easy administration.The effective dose of human oral (to moving chronic hepatitis) is 15-20mg/kg (body weight for humans), and its safety ratio is (12.4-16.8 doubly), and efficient is 68%, shows this medicinal ingredients safe ready.Comprehensive above-mentioned chemistry, the result of pharmacology, confirm Cynanchum otophvllum glucoside third, the glucoside fourth, glucoside penta, glucoside the sixth of the twelve Earthly Branches, glucoside has immunomodulatory and enhancement heptan, the move chronic hepatitis particularly viral to hepatitis has therapeutic action, it is efficient near nuclear class medicine and alpha-interferon combined treatment effect, all at 68% clinical efficacy, and Cynanchum otophvllum antihepatitis drug, toxic side effect is little: other class medicines mostly have bigger toxic side effect, as nucleotide drug, particularly some quite serious toxic side effect appear in the alpha-interferon clinical application, according to Giustina etc. at document Giustina et al:Ital J.Gastroenterol, 26:203, report in 1994, use alpha-interferon treatment chronic viral hepatitis and spirit depressing occurs, autoimmune disorder, diabetes, cardiovascular disorder, various organ failures are died from side effects such as hemolytic anemia, minority.
In addition, Cynanchum otophvllum glucoside third, glucoside fourth, glucoside penta, glucoside the sixth of the twelve Earthly Branches, glucoside have immunocompetence heptan, and chloroform is injected the Meniere pathological model (the dizzy rotation of cavy, ocular ataxy, head are vacillated now to the left, now to the right) that causes in cavy external auditory meatus deep good therapeutic action.Efficient is 85-90%, with control group notable difference is arranged; Cynanchum otophvllum glucoside third or glucoside fourth or glucoside penta or glucoside toxic side effect in the sixth of the twelve Earthly Branches are little, and convenient oral is the anti-Meniere's medicine of effective low toxicity.
Embodiment one:
Get Asclepiadaceae Cynanchum plant Cynanchum otophvllum (Cynanchum otophyllum Schneid) root 10kg, extract 2 times 70 ℃ of heat with chloroform, reclaim dope behind the solvent with the sherwood oil degreasing that refluxes, substrate be drug effect position thick glucoside block 200g, with CCL4-AcoEt (95: 5; 85: 15v/v) extract to such an extent that (95: 5) extract 38 restrains and (85: 15v/v) extract 107.5g successively, the latter uses silica gel column chromatography, (99: 1,98: 2,95: 5,85: 15v/v) gradient elution, every 1000ml were a, receive 44 parts altogether to use CHCL3, CHCL3-MeOH successively.The latter is through silica gel H medium pressure column chromatography and Rp-18 medium pressure column chromatography repeatedly, more respectively with chloroform+methyl alcohol (98: 2 and 95: 5v/v), (70: 30 and 80: 20v/v) the mixed solution wash-out got final product methyl alcohol+water.Get Cynanchum otophvllum glucoside third 640mg, glucoside fourth 1020mg, glucoside penta 520mg, glucoside 560mg in the sixth of the twelve Earthly Branches, glucoside 610mg in heptan respectively.In glucoside third or glucoside fourth or glucoside penta or glucoside the sixth of the twelve Earthly Branches or glucoside heptan and vehicle weight ratio is that 1: 2 ratio adds vehicle starch, makes tablet.
Embodiment two:
Obtain Cynanchum otophvllum glucoside third, glucoside fourth, glucoside penta, glucoside the sixth of the twelve Earthly Branches, glucoside heptan with embodiment one described method, be pressed into the capsule for medicine of packing into behind the powder then and make the Cynanchum otophvllum capsule.
Embodiment three:
Get Asclepiadaceae Cynanchum plant Cynanchum otophvllum (Cynanchum otophyllum Schneid) root 10kg, extract 3 times 65 ℃ of heat with chloroform, reclaim dope behind the solvent with the sherwood oil degreasing that refluxes, substrate be drug effect position block 250g, with CCL4-AcoEt (95: 5; 85: 15v/v) extract to such an extent that (95: 5) extract 42 restrains and (85: 15v/v) extract 117.5g successively, the latter uses silica gel column layer spare, (99: 1,98: 2,95: 5,85: 15v/v) gradient elution, every 1000ml were a, receive 44 parts altogether to use CHCL3, CHCL3-MeOH successively.The latter is through silica gel H medium pressure column chromatography and Rp-18 medium pressure column chromatography repeatedly, more respectively with chloroform+methyl alcohol (98: 2 and 95: 5v/v), (70: 30 and 80: 20v/v) the mixed solution wash-out got final product methyl alcohol+water.Respectively Cynanchum otophvllum glucoside third 660mg, glucoside fourth 1100mg, glucoside penta 530mg, glucoside 560mg in the sixth of the twelve Earthly Branches, glucoside 630mg in heptan, formulation method is routinely made glucoside third or glucoside fourth or glucoside penta or glucoside the sixth of the twelve Earthly Branches or glucoside heptan the nourushing syrup of one of concentration 0.5g/10ml.
Embodiment four:
Get Asclepiadaceae Cynanchum plant Cynanchum otophvllum (Cynanchum otophyllum Schneid) root 10kg, extract 2 times 65 ℃ of heat with ethyl acetate, reclaim dope behind the solvent with the sherwood oil degreasing that refluxes, substrate be drug effect position block 300g, with CCL4-AcoEt (95: 5; 85: 15v/v) extract to such an extent that (95: 5) extract 45 restrains and (85: 15v/v) extract 127.5g successively, the latter uses silica gel column layer spare, (99: 1,98: 2,95: 5,85: 15v/v) gradient elution, every 1000ml were a, receive 44 parts altogether to use CHCL3, CHCL3-MeOH successively.The latter is through silica gel H medium pressure column chromatography and Rp-18 medium pressure column chromatography repeatedly, more respectively with chloroform+methyl alcohol (98: 2 and 95: 5v/v), (70: 30 and 80: 20v/v) the mixed solution wash-out got final product methyl alcohol+water.Respectively Cynanchum otophvllum glucoside third 670mg, glucoside fourth 1200mg, glucoside penta 535mg, glucoside 570mg in the sixth of the twelve Earthly Branches, glucoside 640mg in heptan, be 1: 4 ratio adding vehicle starch in glucoside third or glucoside fourth or glucoside penta or glucoside the sixth of the twelve Earthly Branches or glucoside heptan and vehicle weight ratio, make tablet.
Embodiment five:
Get Asclepiadaceae Cynanchum plant Cynanchum otophvllum (Cynanchum otophyllum Schneid) root 10kg, extract 3 times 70 ℃ of heat with ethyl acetate, reclaim dope behind the solvent with the sherwood oil degreasing that refluxes, substrate be drug effect position block 300g, with CCL4-AcoEt (95: 5; 85: 15v/v) extract to such an extent that (95: 5) extract 45 restrains and (85: 15v/v) extract 127.5g successively, the latter uses silica gel column layer spare, (99: 1,98: 2,95: 5,85: 15v/v) gradient elution, every 1000ml were a, receive 44 parts altogether to use CHCL3, CHCL3-MeOH successively.The latter is through silica gel H medium pressure column chromatography and Rp-18 medium pressure column chromatography repeatedly, more respectively with chloroform+methyl alcohol (98: 2 and 95: 5v/v), (70: 30 and 80: 20v/v) the mixed solution wash-out got final product methyl alcohol+water.Respectively Cynanchum otophvllum glucoside third 670mg, glucoside fourth 1200mg, glucoside penta 535mg, glucoside 570mg in the sixth of the twelve Earthly Branches, glucoside 640mg in heptan, be pressed into the capsule for medicine of packing into behind the powder heptan by glucoside third or glucoside fourth or glucoside penta or glucoside the sixth of the twelve Earthly Branches or glucoside and make the Cynanchum otophvllum capsule.
Embodiment six:
Get Asclepiadaceae Cynanchum plant Cynanchum otophvllum (Cynanchum otophyllum Schneid) root 10kg, extract 3 times 60 ℃ of heat with ethyl acetate, reclaim dope behind the solvent with the sherwood oil degreasing that refluxes, substrate be drug effect position 250 grams, with CCL4-AcoEt (95: 5; 85: 15v/v) extract to such an extent that (95: 5) extract 38 restrains and (85: 15v/v) extract 107.5g successively, the latter uses silica gel column layer spare, (99: 1,98: 2,95: 5,85: 15v/v) gradient elution, every 1000ml were a, receive 44 parts altogether to use CHCL3, CHCL3-MeOH successively.The latter is through silica gel H medium pressure column chromatography and Rp-18 medium pressure column chromatography repeatedly, more respectively with chloroform+methyl alcohol (98: 2 and 95: 5v/v), (70: 30 and 80: 20v/v) the mixed solution wash-out got final product methyl alcohol+water.Respectively Cynanchum otophvllum glucoside third 660mg, glucoside fourth 1100mg, glucoside penta 530mg, glucoside 560mg in the sixth of the twelve Earthly Branches, glucoside 630mg in heptan, formulation method is routinely made glucoside third or glucoside fourth or glucoside penta or glucoside the sixth of the twelve Earthly Branches or glucoside heptan the nourushing syrup of one of concentration 0.2g/ml.
Embodiment seven:
Get Asclepiadaceae Cynanchum plant Cynanchum otophvllum (Cynanchum otophyllum Schneid) root 10kg, extract 2 times 70 ℃ of heat with chloroform, reclaim dope behind the solvent with the sherwood oil degreasing that refluxes, substrate be drug effect position thick glucoside block 200g, with CCL4-AcoEt (95: 5; 85: 15v/v) extract to such an extent that (95: 5) extract 38 restrains and (85: 15v/v) extract 107.5g successively, the latter uses silica gel column layer spare, (99: 1,98: 2,95: 5,85: 15v/v) gradient elution, every 1000ml were a, receive 44 parts altogether to use CHCL3, CHCL3-MeOH successively.The latter is through silica gel H medium pressure column chromatography and Rp-18 medium pressure column chromatography repeatedly, more respectively with chloroform+methyl alcohol (98: 2 and 95: 5v/v), (70: 30 and 80: 20v/v) the mixed solution wash-out got final product methyl alcohol+water.Respectively Cynanchum otophvllum glucoside third 640mg, glucoside fourth 1020mg, glucoside penta 520mg, glucoside 560mg in the sixth of the twelve Earthly Branches, glucoside 610mg in heptan, be 1: 2 ratio adding vehicle starch in glucoside third or glucoside fourth or glucoside penta or glucoside the sixth of the twelve Earthly Branches or glucoside heptan and vehicle weight ratio, make tablet.
Embodiment eight:
Obtain Cynanchum otophvllum glucoside third, glucoside fourth, glucoside penta, glucoside the sixth of the twelve Earthly Branches or glucoside heptan with embodiment seven described methods, be pressed into the capsule for medicine of packing into behind the powder then and make the Cynanchum otophvllum capsule.
Embodiment nine:
Get Asclepiadaceae Cynanchum plant Cynanchum otophvllum (Cynanchum otophyllum Schneid) root 10kg, extract 3 times in 65C heat with chloroform, reclaim dope behind the solvent with the sherwood oil degreasing that refluxes, substrate be drug effect position block 250g, with CCL4-AcoEt (95: 5; 85: 15v/v) extract to such an extent that (95: 5) extract 42 restrains and (85: 15v/v) extract 117.5g successively, the latter uses silica gel column layer spare, (99: 1,98: 2,95: 5,85: 15v/v) gradient elution, every 1000ml were a, receive 44 parts altogether to use CHCL3, CHCL3-MeOH successively.The latter is through silica gel H medium pressure column chromatography and Rp-18 medium pressure column chromatography repeatedly, more respectively with chloroform+methyl alcohol (98: 2 and 95: 5v/v), (70: 30 and 80: 20v/v) the mixed solution wash-out got final product methyl alcohol+water.Respectively Cynanchum otophvllum glucoside third 660mg, glucoside fourth 1100mg, glucoside penta 530mg, glucoside 560mg in the sixth of the twelve Earthly Branches, glucoside 630mg in heptan, formulation method is routinely made glucoside third or glucoside fourth or glucoside penta or glucoside the sixth of the twelve Earthly Branches or glucoside heptan the nourushing syrup of one of 0.5g/10ml.
Embodiment ten:
Get Asclepiadaceae Cynanchum plant Cynanchum otophvllum (Cynanchum otophyllum Schneid) root 10kg, extract 2 times 65 ℃ of heat with ethyl acetate, reclaim dope behind the solvent with the sherwood oil degreasing that refluxes, substrate be drug effect position block 300g, with CCL4-AcoEt (95: 5; 85: 15v/v) extract to such an extent that (95: 5) extract 45 restrains and (85: 15v/v) extract 127.5g successively, the latter uses silica gel column layer spare, (99: 1,98: 2,95: 5,85: 15v/v) gradient elution, every 1000ml were a, receive 44 parts altogether to use CHCL3, CHCL3-MeOH successively.The latter is through silica gel H medium pressure column chromatography and Rp-18 medium pressure column chromatography repeatedly, more respectively with chloroform+methyl alcohol (98: 2 and 95: 5v/v), (70: 30 and 80: 20v/v) the mixed solution wash-out got final product methyl alcohol+water.Respectively Cynanchum otophvllum glucoside third 670mg, glucoside fourth 1200mg, glucoside penta 535mg, glucoside 570mg in the sixth of the twelve Earthly Branches, glucoside 640mg in heptan, be 1: 4 ratio adding vehicle starch in glucoside third or glucoside fourth or glucoside penta or glucoside the sixth of the twelve Earthly Branches or glucoside heptan and vehicle weight ratio, make tablet.
Embodiment 11:
Get Asclepiadaceae Cynanchum plant Cynanchum otophvllum (Cynanchum otophyllum Schneid) root 10kg, extract 3 times 70 ℃ of heat with ethyl acetate, reclaim dope behind the solvent with the sherwood oil degreasing that refluxes, substrate be drug effect position block 300g, with CCL4-AcoEt (95: 5; 85: 15v/v) extract to such an extent that (95: 5) extract 45 restrains and (85: 15v/v) extract 127.5g successively, the latter uses silica gel column layer spare, (99: 1,98: 2,95: 5,85: 15v/v) gradient elution, every 1000ml were a, receive 44 parts altogether to use CHCL3, CHCL3-MeOH successively.The latter is through silica gel H medium pressure column chromatography and Rp-18 medium pressure column chromatography repeatedly, more respectively with chloroform+methyl alcohol (98: 2 and 95: 5v/v), (70: 30 and 80: 20v/v) the mixed solution wash-out got final product methyl alcohol+water.Respectively Cynanchum otophvllum glucoside third 670mg, glucoside fourth 1200mg, glucoside penta 535mg, glucoside 570mg in the sixth of the twelve Earthly Branches, glucoside 640mg in heptan, press glucoside third or glucoside fourth or glucoside penta or glucoside the sixth of the twelve Earthly Branches or glucoside heptan, be pressed into the capsule for medicine of packing into behind the powder and make the Cynanchum otophvllum capsule.
Embodiment 12:
Get Asclepiadaceae Cynanchum plant Cynanchum otophvllum (Cynanchum otophyllum Schneid) root 10kg, extract 3 times 60 ℃ of heat with ethyl acetate, reclaim dope behind the solvent with the sherwood oil degreasing that refluxes, substrate be drug effect position 200 grams, with CCL4-AcoEt (95: 5; 85: 15v/v) extract to such an extent that (95: 5) extract 38 restrains and (85: 15v/v) extract 107.5g successively, the latter uses silica gel column layer spare, (99: 1,98: 2,95: 5,85: 15v/v) gradient elution, every 1000ml were a, receive 44 parts altogether to use CHCL3, CHCL3-MeOH successively.The latter is through silica gel H medium pressure column chromatography and Rp-18 medium pressure column chromatography repeatedly, more respectively with chloroform+methyl alcohol (98: 2 and 95: 5v/v), (70: 30 and 80: 20v/v) the mixed solution wash-out got final product methyl alcohol+water.Respectively Cynanchum otophvllum glucoside third 660mg, glucoside fourth 1100mg, glucoside penta 530mg, glucoside 560mg in the sixth of the twelve Earthly Branches, glucoside 630mg in heptan, formulation method is routinely made glucoside third or glucoside fourth or glucoside penta or glucoside the sixth of the twelve Earthly Branches or glucoside heptan the nourushing syrup of one of concentration 0.2g/ml.
The physico-chemical property in the glucoside third that makes in the foregoing description, glucoside fourth, glucoside penta, glucoside the sixth of the twelve Earthly Branches, glucoside heptan is as follows:
Glucoside third: be white powder, it is 630 that fusing point 159-162 degree, FAB-MS are measured its molecular weight; UV λ MeoH/max, nm (log ε): 204 (4.2), 259 (4.1); Infrared IRv KBr/maxem13430 (OH), 3400,2930,1690 (c-o) .1585,1505 (aromatic rings), 1475 (CH3), 1270,1230,1195,1160,1075,1045 (c-o-o), 975,850,770, nucleus magnetic hydrogen spectrum (CDC13) 1.05 (3H, S.ZC19-CH3), 1.23 (3H, d.J=6.4Hz. sugar C6-CH3), 1.50 (3H, S.C18-CH3), 2.08 (3H, S, C21-CH3), 3.52 (1H, S.C3-aH), 4.70 (1H, dd.J1=11.2Hz, J2=4.4Hz, C12-H), 4.87 (1H, d.J=8Hz, sugar C1-H), 5.3 (1H, S.Ce-H), 6.75, (7.73 4H, dd, J=8.76Hz, 4H on the 1.4-=substituted benzene ring).
The glucoside fourth: be white powder, fusing point 154-157 ℃, it is 1056 that FAB-MS measures its molecular weight; UV λ KBr/max, nm (log e): 206 (4.2), 219 (4.3), 281 (3.2); Infrared IRVKBr/max cm13450 (OH), 2960,2930 (1440) (OCH3), 1700,1635 (C=CH-CO), 1520,1370,1350,1220,1160,1080,1050 (C-O-O), 1000,980,910,860,750; Nucleus magnetic hydrogen spectrum (CDC13) 1.06 (6H, d.J=7Hz ,-CH-CH3), 1.13 (3H, S.C19-CH3), 1.19 (3H, d, J=6Hz, sugar C6-CH3), 1.21 (3H, d.J=6Hz, sugared C6-CH3), (1.25 3H, d, J=8Hz, sugared C6-CH3), 1.50 (3H, S, C18-CH3), 2.11 (3H, S.C7-CH3), 2.17 (6H, S. sugar C3-OCH3), 4.31 (1H, m, C3-aH), 4.44 (4H, dd, J=8Hz, 4 sugared C1-H), 4.56 (1H, dd/J=9Hz.C12-aH), (4.84 4H, t.J=10Hz, 4 sugared C4-H), 5.34 (1H, br.C6-H), 5.53 (1H, S.C2-H).
Glucoside penta: be the unformed powder of white, molecular formula C48H76O16, fusing point 159-162 ℃, MS (m/z): 908 (m+), 633,517,451,359,264,173,145,127.1H-NMR (C5D5N): 6.45 (1H, brd, OH), 5.85 (1H, S, 2-H), 5.45 (ah, S, 6-H), 3.56,3.45 (each 3H, S, 2XOCH3), 2.60 (3H, S, 21-CH3), 2.26 (3H, S, 7-CH3), 1.97 (3H, S, 19-CH3), 1.55,1.43,1.35 (each 3H, d, J=6.0Hz, sugar 6-CH3), 1.30 (3H, S, 18-CH3), 0.93 (6H, d, 5,6-CH3).
Glucoside the sixth of the twelve Earthly Branches: be the unformed powder of white, fusing point 159-162 ℃, molecular formula C48H76O16; MS (m/z): 908,798,716,619,511,359,281,188,127.H-NMR (CDCl3): 5.39 (1H, S, 2-H), 5.23 (1H, S, 6-H), 4.80,4.69 (each 1H, brd, J=9.4Hz, sugared 1-H), 4.39 (2H, brd, J=10.6Hz, 12d-H, sugared 1-H), 4.11 (1H, brs, cymarose 3-H), (3.76 1H, dq, sugared 5-H), 3.31,3.24 (each 3H, S, 2XOCH3), 2.97 (1H, t, oleandrose 4-H), 2.04 (3H, S, 21-CH3), 1.94 (3H, S, 7-CH3), 1.34 (3H, S, 19-CH3), 1.18 (3H, d, J=6.0Hz, sugared 6-CH3), 1.10 (6H, d, J=6.0Hz, sugared 6-CH3), 1.01 (3H, S, 18-CH3), 0.94 (6H, d, J=7.0Hz, 5,6-CH3).
Glucoside heptan: be the unformed powder of white, fusing point 159-162 ℃, molecular formula C44H70 O 16, MS (m/z): 854 (m+), 794 (M_HOAc), 710 (M+-145+H), 643,582,522,419,357,325,283,255,145,127.H-NMR (C5D5N): 6.53 (1H, S, OH), 6.25 (1H, S, 6-H), 4.95 (1H, dd, J=11.5,3.7Hz, 12a-H), 4.76 (1H, brd, J=8.3Hz, sugar 1-aH), 4.20 (2H, dq, two cymarose 5-H), 4.09, (4.05 each 1H, brd, J=2.6Hz, two cymarose 3-H), 3.85 (1H, m, 3a-H), 3.61,3.57,3.47 (each 3H, S, 3XOCH3), 2.46 (3H, S, 21-CH3), 2.05 (3H, S, OAc), 1.93 (3H, S, 18-CH3), 1.57,1.41,1.38 (each 3H, d, J=6.1Hz, sugared 6-CH3), 1.31 (3H, S, 19-CH3).
Claims (7)
1, general formula (I) is
Five C21 steroidal glycoside compounds, compound 1 in the formula (Cynanchum otophvllum glucoside third)
Compound 2 (Cynanchum otophvllum glucoside fourth)
Compound 3 (Cynanchum otophvllum glucoside penta)
Compound 4 (Cynanchum otophvllum glucoside the sixth of the twelve Earthly Branches)
Compound 5 (Cynanchum otophvllum glucoside heptan)
R2＝CH
3CO
-
2, the preparation method of the general formula compound of claim 1 is characterized in that adopting vegetable chemistry method extraction separation from the plant Cynanchum otophvllum.
3, as the preparation method of the said general formula compound of claim 2, it is characterized in that getting Asclepiadaceae Cynanchum plant Cynanchum otophvllum (Cynanchum otophyllum Schneid) root, extract 2-3 time 60-70 ℃ of heat with chloroform, reclaim dope behind the solvent with the sherwood oil degreasing that refluxes, getting substrate is the thick glucoside block in drug effect position, uses CCLA-AcoEt (95: 5; 85: 15v/v) extract successively (95: 5v/v) extract and (85: 15v/v) extract, the latter is through silica gel H medium pressure column chromatography and Rp-18 medium pressure column chromatography repeatedly, more respectively with chloroform+methyl alcohol (98: 2 and 95: 5v/v), (70: 30 and 80: 20v/v) the mixed solution wash-out got final product methyl alcohol+water.
4, as the preparation method of the said general formula compound of claim 2, it is characterized in that getting Asclepiadaceae Cynanchum plant Cynanchum otophvllum (Cynanchum otophyllum Schneid) root, extract 2-3 time 60-70 ℃ of heat with ethyl acetate, reclaim dope behind the solvent with the sherwood oil degreasing that refluxes, getting substrate is the thick glucoside block in drug effect position, uses CCL4-AcoEt (95: 5; 85: 15v/v) extract successively (95: 5v/v) extract and (85: 15v/v) extract, the latter is through silica gel H medium pressure column chromatography and Rp-18 medium pressure column chromatography repeatedly, more respectively with chloroform+methyl alcohol (98: 2 and 95: 5v/v), (70: 30 and 80: 20v/v) the mixed solution wash-out got final product methyl alcohol+water.
5, the application of the general formula compound of claim 1 in the preparation Antihepatitis medicament.
6, application according to claim 5 is characterized in that the application in the medicine of preparation treatment chronic persistent hepatitis and chronic hepatitis disease.
7, the application of the general formula compound of claim 1 in preparation anti-Meniere's medicine.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1292754C (en) * | 2004-11-09 | 2007-01-03 | 云南白药集团股份有限公司 | Application of C21 steroid glycoside in pharmacy |
| CN101953864A (en) * | 2010-09-29 | 2011-01-26 | 中国人民解放军军事医学科学院毒物药物研究所 | Cynanchum otophyllum aglycone and medical application of extractive containing same |
| CN101357150B (en) * | 2008-08-29 | 2011-07-20 | 贵州大学 | Method for separating and purifying Qingyangshenylycosides from Qingyangsheny |
| CN102550611A (en) * | 2011-01-04 | 2012-07-11 | 昆明华地丰润生物科技有限公司 | Plant rodenticide and preparation method thereof |
| CN102924554A (en) * | 2012-10-29 | 2013-02-13 | 中国科学院昆明植物研究所 | C-3,11,12,20-tetrasubstituted C-21 steroid derivative and its pharmaceutical composition and use in medicine |
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Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH07116216B2 (en) * | 1986-09-04 | 1995-12-13 | メクト株式会社 | Method for producing sialosil cholesterol |
| CN1019935C (en) * | 1987-03-09 | 1993-02-24 | 中国人民解放军空军桂林医院 | Method for extracting sweetening agent from momordica grosvenori |
| DE4001895A1 (en) * | 1990-01-23 | 1991-07-25 | Harrier Gmbh | PRODUCTION OF PARTICULAR STEROID GLYCOSIDES |
| TW479061B (en) * | 1993-12-24 | 2002-03-11 | Mitsubishi Chem Corp | Sialic acid derivatives |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1292754C (en) * | 2004-11-09 | 2007-01-03 | 云南白药集团股份有限公司 | Application of C21 steroid glycoside in pharmacy |
| CN101357150B (en) * | 2008-08-29 | 2011-07-20 | 贵州大学 | Method for separating and purifying Qingyangshenylycosides from Qingyangsheny |
| CN101953864A (en) * | 2010-09-29 | 2011-01-26 | 中国人民解放军军事医学科学院毒物药物研究所 | Cynanchum otophyllum aglycone and medical application of extractive containing same |
| CN102550611A (en) * | 2011-01-04 | 2012-07-11 | 昆明华地丰润生物科技有限公司 | Plant rodenticide and preparation method thereof |
| CN102924554A (en) * | 2012-10-29 | 2013-02-13 | 中国科学院昆明植物研究所 | C-3,11,12,20-tetrasubstituted C-21 steroid derivative and its pharmaceutical composition and use in medicine |
| CN102924554B (en) * | 2012-10-29 | 2015-07-22 | 中国科学院昆明植物研究所 | C-3,11,12,20-tetrasubstituted C-21 steroid derivative and its pharmaceutical composition and use in medicine |
| CN110128496A (en) * | 2019-06-06 | 2019-08-16 | 贵州师范大学 | It accuses up to pavilion astragalin composition and its preparation method and application |
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