CN102146057B - C19-diterpenoid alkaloid, preparation method of C19-diterpenoid alkaloid, medical composite with C19-diterpenoid alkaloid used as active ingredient and application of medical composite - Google Patents
C19-diterpenoid alkaloid, preparation method of C19-diterpenoid alkaloid, medical composite with C19-diterpenoid alkaloid used as active ingredient and application of medical composite Download PDFInfo
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Abstract
The invention relates to C19-diterpenoid alkaloid which has cardiotonic and hart failure-resistance activities and is shown in the formula I, pharmaceutically acceptable salts of the C19-diterpenoid alkaloid, preparation methods of the C19-diterpenoid alkaloid and the salts, and medicines containing the C19-diterpenoid alkaloid and the salts, as well as application of the C19-diterpenoid alkaloid and the salts as medicines with cardiotonic and hart failure-resistance effects. In the formula I, R1 is hydrogen, hydroxyl or methoxyl; R2 is hydrogen or methyl; and R3, R4 and R5 represent hydrogen or hydroxyl.
Description
Technical field:
The present invention relates to have the C of pharmaceutical use
19-diterpene alkaloid N-goes ethyl aconine alkali, the peaceful alkali of north crow, N-to remove the peaceful alkali of ethyl time crow and peaceful alkali of different Dare and homologue thereof.They divide is raw material in monkshood or with the napelline, semi-synthetic preparation.
The invention still further relates to two kinds of C
19Peaceful alkali of crow and the peaceful alkali of time crow in-the diterpene alkaloid.They are raw material to divide the napelline in the plant rhizome of Chinese monkshood, are prepared from semi-synthetic.
Simultaneously, the preparation method of above-claimed cpd and be the pharmaceutical composition of activeconstituents with this compound is disclosed.The invention belongs to natural drug extracts active ingredients and preparing technical field.
Background technology:
The Chinese medicine monkshood is the processed goods of the sub-root of the ranunculaceae plant rhizome of Chinese monkshood (Aconitum carmichaeli Debx.).Have recuperating deplered YANG and rescuing the patient from collapse, it is supporing yang to mend fire, by wind-cold damp pathogen, and the antalgic function.Be used for the yang depletion collapse, the cold weak pulse of limb, impotence, the palace is cold, trusted subordinate's crymodynia, the cold of insufficiency type is vomited and diarrhoea, cold sensation of the genitalia oedema, deficiency of yang diseases caused by external factors, arthralgia due to cold-dampnes (Chinese Pharmacopoeia, version was an one in 2000, the 150th page).At China's monkshood aboundresources, it is wider to distribute, and is widely used in clinical as famous Chinese medicine.
Discover cardiac stimulant composition in the monkshood have racemization go the black alkali of first (dl-demethylcoclaurine) (T.Kosuge et al, Chem.Pharm.Bull, 1976,24:176-178), methyl chloride dopamine (coryneine chloride) (C.Lammp; Plata Medica, 1979,35:150-), salsolidine alkali (salsoline) (Chen Dihua etc.; Acta Pharmaceutica Sinica, 1982,17:792-794), uracil (uracil) (Han Gongyu etc.; Research and development of natural products, 1997,9 (3): 30-34) and monkshood aglycone (fuzinoside) (Xu Dongming etc.; Chinese herbal medicine, 2004,35:964-966; Chinese invention patent, 2006, CN100386338C).But, still do not have and report that the diterpene alkaloid compound of from monkshood, telling has cardiotonic.In recent years, the method that we adopt cardiac activity to follow the trail of is discovered, from monkshood, tells the Compound C with significant cardiac stimulant and anti-heart failure effect
19-diterpene alkaloid N-goes ethyl aconine alkali, Bei Wuning, N-to remove the peaceful alkali of ethyl time crow, the peaceful alkali of different Dare, the peaceful alkali of middle crow and the peaceful alkali of inferior crow.But because these alkaloids are lower in the monkshood, we have studied with the higher napelline of content in the product rhizome of Chinese monkshood of Xinjiang again is raw material, and the semi-synthetic N-of preparation goes the method for the peaceful alkali of ethyl time crow, the peaceful alkali of middle crow and the peaceful alkali of inferior crow.
Summary of the invention:
The invention provides a kind of pharmaceutical use C that has
19-diterpene alkaloid N-goes ethyl aconine alkali, Bei Wuning, N-to remove the peaceful alkali of ethyl time crow and peaceful alkali of different Dare and homologue thereof.
The present invention also provides the method for from the monkshood and the rhizome of Chinese monkshood, extracting above-mentioned substance.
It is raw material that the present invention also provides with the napelline, prepares the method that N-removes ethyl aconine alkali, the peaceful alkali of middle crow and the peaceful alkali of inferior crow, is suitable for suitability for industrialized production.
It is the pharmaceutical composition of activeconstituents that the present invention further provides with the above-claimed cpd, is used to prepare the medicine of cardiac stimulant and anti-heart failure.
Compound of the present invention has following general structural formula (I):
R wherein
1Be hydrogen, hydroxyl or methoxyl group; R
2Be hydrogen or methyl; R
3, R
4And R
5Be hydrogen or hydroxyl.
C with cardiac stimulant and anti-heart failure effect of the present invention
19Peaceful alkali of crow and the peaceful alkali of time crow during-diterpene alkaloid compound comprises.
The present invention provides following technological method:
(1) extraction separation N-removes ethyl aconine alkali (XII) and the peaceful alkali of different Dare (XIII) from the monkshood or the rhizome of Chinese monkshood.
(2) Wu Ting is a raw material to the north of, through hydrolysis preparation north crow peaceful (XIV).
(3) with the napelline be raw material, through full acetylated, N-go to ethylize, N-methylates and hydrolysis, the peaceful alkali (II) of crow in the preparation.
(4) with the napelline be raw material, full acetylated through chloro, elimination, hydrogenation, N-goes to ethylize, and N-methylates and hydrolysis, the peaceful alkali (III) of preparation time crow.
(5) with the napelline be raw material, full acetylated through chloro, elimination, hydrogenation, N-goes to ethylize and hydrolysis, and preparation N-removes the peaceful alkali (XV) of ethyl time crow.
The method that the present invention extraction separation N-from the monkshood or the rhizome of Chinese monkshood removes ethyl aconine alkali and the peaceful alkali of different Dare may further comprise the steps:
(1) monkshood or rhizome of Chinese monkshood pulverizing back boiling are got extracting solution;
(2) extracting solution alcohol precipitation, filtration, filtrate decompression concentrate, and get solid extract;
(3) extract with water dissolution, alkalization after, n-butanol extraction reclaims solvent, n-butanol extract;
(4) n-butanol extract separates through silica gel column chromatography, and different ratios chloroform-methanol mixed solvent wash-out gets required compound.
Method therefor of the present invention, simple, be suitable for enlargement of scale, be easy to realize the suitability for industrialized production purpose.
The method of the peaceful alkali of crow in the present invention's preparation may further comprise the steps:
1) under the Catalyzed by p-Toluenesulfonic Acid, napelline and Acetyl Chloride 98Min. or aceticanhydride room temperature reaction 6-12 hour get solids after well-established law is handled, and 95% alcohol crystal gets compound 3,13,15-triacetyl napelline (V);
2) 3,13,15-triacetyl napelline mixes with 3 normal N-bromo-succinimides and 20-40 times of Glacial acetic acid min. 99.5, and room temperature reaction 2-6 hour, concentrating under reduced pressure got the white solid thing.Through silica gel column chromatography (sherwood oil: acetone=2: 1), get N-and remove ethyl-3,13,15-triacetyl napelline (VI);
3) N-removes ethyl 3,13, and 15-triacetyl napelline mixes with normal methyl iodide, and THF is a solvent, and room temperature reaction 2-4 hour, decompressing and extracting got the white solid thing.(sherwood oil: ETHYLE ACETATE (3: 1) gets 3,13, black alkali (VII) in the 15-triacetyl through silica gel column chromatography;
4) 3,13, black alkali and 5% sodium hydroxide hydrolysis in the 15-triacetyl, ambient temperature overnight, decompressing and extracting gets the white solid thing.Through silica gel column chromatography (chloroform-methanol that ammoniacal liquor is saturated, 95: 1), get the peaceful alkali (II) of crow in the compound.
The present invention prepares time method of the peaceful alkali of crow, may further comprise the steps:
(1) with the THF is solvent, with napelline and normal SOCl
2Mix, ambient temperature overnight reaction 1-3 hour, decompressing and extracting gets the white solid thing.Add again ETHYLE ACETATE-95% ethanol-Glacial acetic acid min. 99.5 (3: 3: 1, w/w) after the mixed solution, under the catalysis of palladium carbon, normal pressure hydrogenation 1-3 days.Decompressing and extracting gets the white solid thing.Through 95% alcohol crystal, get deoxyaconitine (VIII);
(2) under the Catalyzed by p-Toluenesulfonic Acid, deoxyaconitine mixes with 1.5 normal Acetyl Chloride 98Min.s or aceticanhydride, stirs, and room temperature reaction spends the night, and decompressing and extracting gets the white solid thing.Use 95% alcohol crystal again, get compound 13,15-diacetyl deoxyaconitine) (IX);
(3) 13,15-diacetyl deoxyaconitines mix with 3 equivalent N-bromo-succinimides and 20-40 times of Glacial acetic acid min. 99.5, and room temperature reaction 2-6 hour, concentrating under reduced pressure got the white solid thing.Through silica gel column chromatography (sherwood oil-acetone, 2: 1), compound N-go ethyl-13,15-diacetyl deoxyaconitine (X);
(4) N-removes ethyl-13, and 15-diacetyl deoxyaconitine mixes with the equivalent methyl iodide, and THF is a solvent, and room temperature reaction 2-4 hour, decompressing and extracting got the white solid thing.Through silica gel column chromatography (petroleum ether-ethyl acetate, 3: 1), get compound 13,15-diacetyl time black alkali (XI);
(5) 13,15-diacetyl time black alkali adds 5% sodium hydroxide solution, ambient temperature overnight, well-established law handle the white solid thing.Silica gel column chromatography (chloroform-methanol that ammoniacal liquor is saturated, 95: 1) gets the peaceful alkali (III) of compound time crow.
The present invention prepares the method that N-removes the ethyl time peaceful alkali of crow, may further comprise the steps:
(1) with the napelline be raw material, according in the peaceful alkali method of above-mentioned preparation time crow 1.-3. step prepare N-and remove ethyl-13,15-diacetyl deoxyaconitine (X);
(2) N-removes ethyl-13, and 15-diacetyl deoxyaconitine adds the 5%NaOH methanol solution, ambient temperature overnight, well-established law handle the white solid thing.Silica gel column chromatography (the saturated chloroform-methanol of ammoniacal liquor, 95: 1) wash-out gets required compound N-and removes the peaceful alkali (XV) of ethyl time crow.
The peaceful alkali of crow, the peaceful alkali of inferior crow and N-go the peaceful alkali of ethyl time crow through spectroscopic analysis in above-mentioned, have following physicochemical property respectively:
The peaceful alkali of middle crow: white amorphous powder, specific optical rotation does
Molecular formula is C
24H
39NO
9Soluble in water, be insoluble to propyl alcohol, chloroform.Through comprising spectroscopic analysis such as two-dimentional nuclear-magnetism fully, confirm that its structural formula is (II).The spectroscopic data of the peaceful alkali of middle crow is following:
IR(KBr)cm
-1：3424，2932，2892，2821，1639，1453，1106；
1H NMR (CDCl
3) δ: 3.07 (1H, dd, J=8.4,6.0Hz, H-1), 3.75 (1H, m, H-3); 4.15 (1H, d, J=6.8Hz, H-6), 3.93 (1H, d, J=5.2Hz, H-14 β); 4.49 (1H, t, J=5.6Hz, H-15), 3.19 (1H, d, J=6.0Hz, H-16); 2.91 (1H, s, H-17), 3.65,3.74 (each 1H, ABq, J=9.6Hz, H
2-18), 2.48,2.79 (each 1H, ABq, J=11.2Hz, H
2-19), 2.34 (3H, s, NCH
3), 3.26 (3H, s, OCH
3-1), 3.34 (3H, s, OCH
3-6), 3.64 (3H, s, OCH
3-16), 3.32 (3H, s, OCH
3-18);
13C?NMR(CDCl
3)δ：82.6(C-1)，33.8(C-2)，71.5(C-3)，43.4(C-4)，46.5(C-5)，83.2(C-6)，46.4(C-7)，78.8(C-8)，48.8(C-9)，41.6(C-10)，50.1(C-11)，36.9(C-12)，76.3(C-13)，78.7(C-14)，81.5(C-15)，90.7(C-16)，62.5(C-17)，76.8(C-18)，49.8(C-19)，42.5(NCH
3)，56.3(OCH
3-1)，57.9(OCH
3-6)，61.3(OCH
3-16)，59.1(OCH
3-18)；
ESI-MS?m/z(％)：486[M+H](100)；
HR-MS: it is 486.2699 that quasi-molecule measures definite value, and calculated value is: 486.2624.
The peaceful alkali of inferior crow: white amorphous powder, specific optical rotation does
Molecular formula is C
24H
39NO
8Soluble in water, be insoluble to acetone, chloroform.Through comprising spectrographic technique such as two-dimentional nuclear-magnetism fully, confirm that its structural formula is (III).
The spectroscopic data of the peaceful alkali of inferior crow is following:
IR(KBr)cm
-1：3442，2925，2820，1635，1456，1111；
1H?NMR(CDCl
3)δ：3.12(1H，t，J＝7.2Hz，H-1)，4.12(1H，d，J＝7.2Hz，H-6)，3.82(1H，d，J＝4.8Hz，H-14β)，4.42(1H，d，J＝6.0Hz，H-15)，3.05(1H，d，J＝6.0Hz，H-16)，2.93(1H，s，H-17)，3.65(1H，ABq，J＝8.4Hz，H-18)，2.76(1H，ABq，J＝10.8Hz，H-19)，2.35(3H，s，NCH
3)，3.25(3H，s，OCH
3-1)，3.33(3H，s，OCH
3-6)，3.59(3H，s，OCH
3-16)，3.28(3H，s，OCH
3-18)；
13C?NMR(CDCl
3)δ：84.3(C-1)，25.1(C-2)，32.5(C-3)，38.8(C-4)，46.9(C-5)，83.1(C-6)，46.7(C-7)，78.0(C-8)，48.9(C-9)，41.9(C-10)，50.0(C-11)，37.3(C-12)，76.4(C-13)，78.5(C-14)，81.1(C-15)，91.7(C-16)，62.9(C-17)，80.2(C-18)，56.7(C-19)，41.6(NCH
3)，54.8(OCH
3-1)，56.7(OCH
3-6)，60.1(OCH
3-16)，58.1(OCH
3-18)；
ESI-MS?m/z(％)：470[M+H]
+(100)；
HR-MS m/z: it is 470.2749 that quasi-molecule measures definite value, and calculated value is: 470.2675.
N-removes the peaceful alkali of ethyl time crow: white amorphous powder, specific optical rotation does
(c=0.52, methyl alcohol); Molecular formula is C
23H
37NO
8Soluble in water, be insoluble to chloroform, ether.Confirm that through spectrographic technique its structural formula is (XV).N-goes the spectroscopic data of the ethyl time peaceful alkali of crow following:
IR(KBr)cm
-1：3396，2936，2825，1451，1098；
1H?NMR(CDCl
3)δ：4.60(1H，d，J＝6.4Hz)，4.15(1H，d，J＝6.4Hz)，3.87(1H，d，J＝4.4Hz)，3.70(3H，s)，3.35(3H，s)，3.31(3H，s)，3.20(3H，s)，3.12(1H，d，J＝8.0Hz)；
13C?NMR(CDCl
3)δ：91.0(d)，83.5(d)，82.4(d)，81.1(d)，79.9(t)，78.5(s)，78.4(d)，76.2(s)，61.9(d)，59.1(q)，58.1(q)，56.3(q)，55.3(q)，51.5(d)，50.2(s)，49.0(t)，47.7(d)，42.8(d)，41.1(d)，38.7(s)，36.8(t)，28.5(t)，23.0(t)；
ESI-MS?m/z(％)：456[M+H]
+(100)。
Through isolated frog heart, the external cardiac activity screening of isolated rat heart and normal rat heart and the test of heart failure rat heart cardiac activity, above-claimed cpd has cardiac stimulant and anti-heart failure effect.
Following pharmacological testing has confirmed the cardiac stimulant and the anti-heart failure pharmacologically active of The compounds of this invention.
Experimental example 1: peaceful alkali of middle crow and of the provide protection of the peaceful alkali of time crow to the isolated rat heart ischemic damage and reperfusion damage
One, materials and methods
Laboratory animal: the SD rat, male, Test Animal Centre, Academy of Military Medical Sciences, P.L.A provides, conformity certification number, SCXK (army) 2007-0004.
Receive test product: the peaceful alkali of middle crow, molecular weight 485, soluble in water; The peaceful alkali of inferior crow, molecular weight 469, soluble in water, all be formulated as respective concentration before the use with KH liquid.
Positive control drug: the Deslanoside injection liquid, lot number 090409 is produced by Shanghai Xudong Hipu Medicine Co., Ltd, is formulated as respective concentration with KH liquid before using.
Liquid is used in experiment: KH liquid (adds NaCl:13.7918g successively in the 2000ml zero(ppm) water; KCl:0.7008g; CaCl
2: 0.56605g; Anhydrous MgSO
4: 0.2841g; KH
2PO
4: 0.3212g; NaHCO
3: 4.1803g; Glucose:4.3994g.Application of sample is magnetic agitation simultaneously, and is subsequent use).
Laboratory apparatus:
1.Langendorff the isolated heart perfusion device, U.S. BIOPAC company.
2.DC100A polygraph, U.S. BIOPAC company.
Experimental procedure:
1. rat anesthesia is opened chest rapidly, and left hand is mentioned heart gently with tweezers; The right hand is cut off the tissue around superior and inferior vena cava, pulmonary artery, aorta and the heart, and the extraction heart (near the heart aortic root, keep the length of about 0.5-1.0cm; Being equipped with heart cathetrization uses); Move to immediately in the petridish of 4 ℃ of KH liquid of contain, push heart gently intraventricular residue blood is got rid of, clean the heart remained blood.
2. be connected to heart aorta ascending branch on the sleeve pipe of perfusion rapidly, and prick fixing with toe-in.Carry out perfusion to contain oxygen KH liquid immediately.Ppa pulmonary artery pressure 70mm mercury column, perfusion temperature (38 ± 0.5) ℃, flow velocity (11.5 ± 0.5) ml/min.
3. sidewall is cut an osculum in the left atrium, and little latex water pocket is inserted left ventricle through atrioventricular orifice.Be full of ultrapure water in latex water pocket and the conduit that links to each other, the other end of conduit is thoroughly removed inner size bubble through the T joint pressure transducer.Left indoor pressure makes an experiment after stablizing 30min again.
4. write down the coronary flow of each minute heart with graduated cylinder; Pressure transducer signal is led register system through the bridging amplifier input more.Through computer A cq373 software, accomplish signal in real time collection and processing.
5. divide into groups and receive the reagent thing: model group: after containing oxygen KH liquid perfusion 10min, stop irritating 20min, recover to contain oxygen KH liquid perfusion 60min again; Positive controls with tried drug group: wherein positive drug dosage is 10
-12Mol/L is tried in the thing the peaceful alkali of crow and is 10 with the peaceful alkaline agent amount of time crow
-9Mol/L, immediately through receiving reagent thing hole to give the medicine of respective amount, all the other operate same model group when just recovering perfusion, receive the long-pending 1ml that is of reagent object.
6. the filling back is since 10min before each group record stops irritating and again, and every coronary flow (CBF), left ventricular end diastolic at a distance from 10 minutes pressed (LVEDP), the maximum climbing speed (dp/dtmax) of left indoor pressure and heart rate, n=3.
7. data are represented with mean+SD, and the T-test check is taked in data analysis.
Two, the result is shown in table 1-4.
In the table 1. the peaceful alkali of crow and the peaceful alkali of time crow to the influence of CBF (coronary flow) (ml/min, n=3)
Compare with model group,
*P＜0.05,
*P＜0.01
In the table 2. the peaceful alkali of crow and the peaceful alkali of time crow to the influence of dpdtmax (the maximum climbing speed of left indoor pressure) (mmHg/s, n=3)
Compare with model group,
*P＜0.05,
*P＜0.01
In the table 3. the peaceful alkali of crow and the peaceful alkali of time crow to the influence of LVEDP (left ventricular end diastolic presssure) (mmHg, n=3)
Compare with model group,
*P＜0.05,
*P＜0.01
In the table 4. the peaceful alkali of crow and the peaceful alkali of time crow to the influence of heart rate (inferior/min, n=3)
Compare with model group,
*P＜0.05,
*P＜0.01
Isolated rat heart ischemia model is adopted in this experiment, observes to receive peaceful alkali of crow and the influence of the peaceful alkali of time crow to coronary flow, the maximum climbing speed of left indoor pressure, left ventricular end diastolic presssure and heart rate four indices in the test product.
But the blood flow of coronary flow index reflecting myocardium coronary artery.Visible by this experimental result, model group, positive drug control group, the peaceful alkali of middle crow and and the peaceful alkali group administration of crow after, all appears and increase the trend that afterwards reduces earlier, but each administration group of statistical result showed is compared equal unknown significance difference with model group.
The contractile function of the maximum climbing speed index reflection of left indoor pressure cardiac muscle diastole.Compare with model, positive drug control group 10min and 20min time point after administration obviously increase, and statistical comparisons has significant difference; Being tried in the thing the peaceful alkali group 10min after administration of crow all has significant difference to each time point of 40min, and the peaceful alkali group of inferior crow is compared unknown significance difference with model group.
The left ventricular end diastolic presssure index then reflects the diastole conformability of ventricle indirectly.Compare with model group, positive drug control group all obviously reduces to each time point of 60min at 30min, and significant difference is arranged; In the peaceful alkali of crow compare with model group in 20min left ventricular end diastolic presssure index in five time points of 60min significant difference all arranged, peaceful this index of alkali group of inferior crow is in above-mentioned time point unknown significance difference then.
Aspect heart rate, to compare with model group, positive drug control group each time point after administration all obviously slows down, and statistical analysis has significant difference; The peaceful alkali group of middle crow has significant difference at 10min and 20min time point, and the peaceful alkali group of inferior crow has significant difference at the 10min time point.
The prompting of above-mentioned experimental result receives in the test product the peaceful alkali of crow 10
-9Under the mol/L dosage rat ischemia/pour into isolated heart again there is direct increase myocardial contraction, improves the myocardial relaxation function.
Experimental example 2: the peaceful alkali of middle crow is to the cardiotonic of heart failure rat heart
1 materials and methods
1.1 medicine and reagent
The peaceful alkali of middle crow: molecular weight 485, soluble in water.
1.2 animal and test apparatus
Experimental animal: the SD rat, body weight 200-250g, the male and female dual-purpose is provided by Sichuan University's West China medical experiment animal center.
Test apparatus: the BL-420F bio signal is gathered and treatment system: Chengdu Tai Meng science and technology limited Company is produced and is provided.
1.3 TP
The SD rat is divided into 5 groups at random, 4 every group, be respectively the normal control group, model group (saline water 10ml/kg), the peaceful alkali of middle crow big (0.05mg/kg), in (0.0375mgml/kg), little (0.025ml/kg) dose groups.The normal control group is only carried out left ventricular cannulation record left indoor pressure, does not open breast knot and pricks arteria coroaria sinistra, and operation gives saline water 10ml/kg after accomplishing.
Each organizes rats by intraperitoneal injection 10% Chloral Hydrate 0.03ml/kg anesthesia.Be fixed in the mouse plate, it is subcutaneous that ECG electrode is inserted four limbs, leads with the standard II and monitor the animal electrocardiogram(ECG; Operation separates tracheae, bilateral common carotid arteries; The promoting the circulation of qi cannula connects respirator to keep breathing unobstructed; Row left carotid intubate is connected the recording blood pressure curve through sensator with the collection of BL-420F bio signal with treatment system; Be close to left border of sternum and cut off the 3rd or the 4th rib, vertically passivity is separated the upper and lower intercostal muscle, exposes mediastinum and thoracic cavity, left side, visible heart; Passivity is separated pericardium, and near 2mm, arteria coroaria sinistra left anterior descending branch opening part below pulmonary conus and the left auricle of heart intersection, the sewing needle that will be installed with suture line (6/0) penetrates from the left ventricle side, is passed by the pulmonary conus side, is equipped with ligation and uses; Row right carotid intubate is to the left ventricular recording left indoor pressure, and ligation is intubate fixedly; The ligation LADCA is by above-mentioned separated part ligation coronary artery.Whether raise greater than 0.1mv with the ST section, and the change that combines hemodynamic index modeling is successfully to judge the heart failure model; The ligation coronary artery is stablized the pneumoretroperitoneum administration.Before ligation, after the ligation, after the administration heart rate (HR) of 5min, 10min, 20min, 30min, 40min, 50min, 60min, systolic pressure (SP), diastolic pressure (DP), left ventricular systolic pressure (LVSP), left ventricular end diastolic presssure (LVEDP) and left indoor pressure maximum rise and fall off rate (± dp/dtmax).
All data are all represented with mean+SD
after the modeling; Handling the back with modeling previous crops difference adopts the SPSS13.0 statistical software to carry out relatively t check of mean, inspection level α=0.05.
2 test-results
2.1 the peaceful alkali of middle crow is seen table 5 to the result that influences of heart failure rat heart rate (HR).
In the table 5 the peaceful alkali of crow to the influence of heart failure rat heart rate (HR) (x ± s, n=4)
Compare #:P＜0.05, ##:P＜0.01 with normal group; Compare with model group,
*: P＜0.05,
*: P＜0.01;
With heavy dose group ratio, ▲: P＜0.05, ▲ ▲: P＜0.01; Compare with middle dose groups: △: P＜0.05, △ △: P＜0.01.
Table 5 result shows; Compare between peaceful each dose groups of alkali of middle crow; In dose groups stablize HR not make it obvious downtrending remarkable than other dose groups, the heavy dose of group of rise HR effect obvious (P＜0.05) when 20min especially, all the other are respectively organized each time period HR and compare no significant difference.The peaceful alkali of crow has the effect of stable heart failure rat heart rate in the explanation.
2.2 the peaceful alkali of middle crow is seen table 6 to the result that influences of heart failure rat systolic pressure (SBP).
In the table 6 the peaceful alkali of crow to the influence of heart failure rat systolic pressure (SBP) (x ± s, n=4)
Compare #:P＜0.05, ##:P＜0.01 with normal group; Compare with model group,
*: P＜0.05,
*: P＜0.01;
With heavy dose group ratio, ▲: P＜0.05, ▲ ▲: P＜0.01; Compare with middle dose groups: △: P＜0.05, △ △: P＜0.01.
Table 6 result shows, ratio between each dose groups of the peaceful alkali of middle crow, each time period SBP no significant difference of each dosage.The peaceful alkali of crow has the effect of remarkable rise systolic pressure in the explanation to the heart failure rat.
2.3 the peaceful alkali of middle crow is seen table 7 to the result that influences of heart failure rat diastolic pressure (DBP).
In the table 7 the peaceful alkali of crow to the influence of heart failure rat diastolic pressure (DBP) (x ± s, n=4)
Compare #:P＜0.05, ##:P＜0.01 with normal group; Compare with model group,
*: P＜0.05,
*: P＜0.01;
With heavy dose group ratio, ▲: P＜0.05, ▲ ▲: P＜0.01; Compare with middle dose groups: △: P＜0.05, △ △: P＜0.01.
Table 7 result shows that ratio between each dose groups of the peaceful alkali of middle crow, middle dosage resist the DBP reduction when 50min amplitude is better than small dose group (P＜0.05), and all the other respectively organize relatively no significant difference of each time period DBP.The peaceful alkali of crow has the effect that remarkable antagonism heart failure rat diastolic pressure descends in the explanation.
2.4 the peaceful alkali of middle crow is seen table 8 to the result that influences of heart failure rat left ventricular systolic pressure (LVSP).
In the table 8 the peaceful alkali of crow to the influence of heart failure rat left ventricular systolic pressure (LVSP) (x ± s, n=4)
Compare #:P＜0.05, ##:P＜0.01 with normal group; Compare with model group,
*: P＜0.05,
*: P＜0.01;
With heavy dose group ratio, ▲: P＜0.05, ▲ ▲: P＜0.01; Compare with middle dose groups: △: P＜0.05, △ △: P＜0.01.
Table 8 shows, ratio between each dose groups of the peaceful alkali of middle crow, each time period LVSP no significant difference of each dosage.The peaceful alkali of crow has the effect that significantly improves heart failure rat left chamber contractile function in the explanation.
2.5 the peaceful alkali of middle crow presses the result that influences of (LVEDP) to see table 9 to heart failure rat left chamber diastole end.
In the table 9 the peaceful alkali of crow to heart failure rat left chamber diastole end press (LVEDP) influence (x ± s, n=4)
Compare #:P＜0.05, ##:P＜0.01 with normal group; Compare with model group,
*: P＜0.05,
*: P＜0.01;
With heavy dose group ratio, ▲: P＜0.05, ▲ ▲: P＜0.01; Compare with middle dose groups: △: P＜0.05, △ △: P＜0.01.
Table 9 shows, ratio between each dose groups of the peaceful alkali of middle crow, middle dosage 20,60min, low dose of when 30-40min antagonism LVEDP raise than heavy dose more obvious (P＜0.05～0.01), LVEDP no significant difference between all the other each dose groups of each time period.The peaceful alkali of crow has the effect that reduces heart failure rat left chamber diastole end pressure in the explanation.
(+dp/dtmax) the result that influences sees table 10 2.6 the peaceful alkali of middle crow is to the maximum climbing speed of heart failure rat left ventricle isovolumetric phase pressure.
In the table 10 the peaceful alkali of crow to the maximum climbing speed of heart failure rat left ventricle isovolumetric phase pressure (+dp/dtmax) influence (and x ± s, n=8)
Compare #:P＜0.05, ##:P＜0.01 with normal group; Compare with model group,
*: P＜0.05,
*: P＜0.01;
With heavy dose group ratio, ▲: P＜0.05, ▲ ▲: P＜0.01; Compare with middle dose groups: △: P＜0.05, △ △: P＜0.01.
Table 10 shows, ratio between each dosage of the peaceful alkali of middle crow, middle dosage at 50-60min time+dp/dtmax apparently higher than heavy dose group (P＜0.05), at 60min time+dp/dtmax apparently higher than small dose group (P＜0.05).The peaceful alkali of crow has the effect that significantly improves heart failure rat left chamber maximum collapse speed in the explanation.
(result that influences dp/dtmax) sees table 11 2.7 the peaceful alkali of middle crow is to the maximum fall off rate of heart failure rat left ventricle isovolumetric phase pressure.
In the table 11 the peaceful alkali of crow to the maximum fall off rate of heart failure rat left ventricle isovolumetric phase pressure (influence dp/dtmax) (and x ± s, n=4)
Compare #:P＜0.05, ##:P＜0.01 with normal group; Compare with model group,
*: P＜0.05,
*: P＜0.01;
With heavy dose group ratio, ▲: P＜0.05, ▲ ▲: P＜0.01; Compare with middle dose groups: △: P＜0.05, △ △: P＜0.01.
Table 11 shows, ratio between each dosage of the peaceful alkali of middle crow, and the effect of middle dose groups rising-dp/dtmax 20, during 50-60min is apparently higher than heavy dose group (P＜0.05).
3. the comprehensive The above results of conclusion, the peaceful alkali of middle crow can improve the heart function of heart failure rat.Explain that the peaceful alkali of crow has significant cardiac stimulant and anti-heart failure effect in the compound.
The present invention proposes to contain the medicine of compound or pharmaceutically acceptable salt thereof shown in the general formula of the present invention (I).
The present invention also proposes to contain in the The compounds of this invention medicine of the peaceful alkali of crow and time peaceful alkali of crow or their pharmaceutical salts.
Said compound is converted into suitable pharmaceutical dosage form, uses inert additive and vehicle to process medicine where necessary.
The present invention has proposed in the application compound the peaceful alkali of crow simultaneously and has been used for clinical treatment with time peaceful alkali of crow or their pharmaceutical salts as cardiac stimulant and anti-heart failure medicine.
Drug regimen of the present invention can be ointment, gel, paste, sprays, lotion, suspension agent, solvent or emulsion, syrup, particle or the pulvis of activeconstituents in water and non-diluent water.
The medicine of the peaceful alkali of crow and time black peaceful alkali or their pharmaceutical salts in the application The compounds of this invention, the formulation that can adopt is the lid human relations form of administration of habitually practising, for example: ointment, tablet, pill, capsule, suppository, emulsion, input liquid and injection liquid.These preparations use traditional additive and vehicle to make by well-known method.Making medicine thus as required can be by the part, administration such as non-enteron aisle, oral, injection.
The thinner (like particle) that is used for pharmaceutical composition is applicable to form comprising of tablet, drageeing, capsule, pill and injection:
(a) weighting agent is like W-Gum, sugar and silicic acid;
(b) tackiness agent is like derivatived cellulose, alginate, gelatin and Vinylpyrrolidone polymer;
(c) wetting agent is like glycerine;
(d) disintegrating agent is like agar, lime carbonate and sodium hydrogencarbonate;
(e) absorption enhancer is like quaternary ammonium compound;
(f) tensio-active agent is like cetyl alcohol;
(g) absorption carrier is like kaolin and soap clay;
(h) lubricant is like talcum powder, calcium stearate, Magnesium Stearate and solid polyethylene glycol;
(i) pH regulator agent, example hydrochloric acid, sodium hydrogencarbonate, sodium hydroxide;
(j) isotonic agent is like citric acid, Trisodium Citrate BP, lactic acid and Sodium.alpha.-hydroxypropionate;
(k) solubility promoter is like Tegosept M, ethyl p-hydroxybenzoate, tween-80;
(l) inhibitor is like Sodium Pyrosulfite, Sulfothiorine, S-WAT and vitamins C.
By tablet, drageeing, the capsule pill that medicinal compsns of the present invention forms, can hold common dressing, coating and protectiveness matrix, it can contain the lucifuge agent.They can be made into such composition, make they over a period to come with activeconstituents only or preferably special in vivo part discharge.Dressing, coating and protectiveness matrix can be with processing like poly material or wax and so on.
Also available one or more the above-mentioned thinners of activeconstituents are processed the microcapsule packet form together.
Any currently known methods that aforementioned pharmaceutical compositions or production of medicine can be used in this technology carries out, and for example one or more activeconstituentss and one or more mixing diluents is formed medicinal compsns, again compsn is processed medicine.
According to compound shown in the general formula of the present invention (I) and the peaceful alkali of middle crow and the peaceful alkali of time crow as the dosage in the clinical medicine, preferably in scope, preferably in 0.01-5 milligram/kg body weight scope with 0.0001 to 5 milligram/kg body weight.
Pharmacological evaluation through the present invention did finds that compound shown in the general formula (I) and the peaceful alkali of middle crow have cardiac stimulant and anti-heart failure effect with time peaceful alkali of crow or their pharmaceutical salts.This effect can be used as cardiac stimulant and anti-heart failure drug use clinically.
Embodiment:
Embodiment 1:N-goes the preparation of ethyl aconine alkali (XII)
Take by weighing monkshood 22kg, with 3 times of calorimetric water extraction 3 times, behind extracting solution 75% ethanol sedimentation, the supernatant concentrating under reduced pressure, extract 175g.Use water dissolution, ammoniacal liquor is transferred pH 10, ether defatting.Water layer is used n-butanol extraction, and extraction liquid is drained, and gets n-butanol extract 10g.Silica gel (Haiyang Chemical Plant, Qingdao, down together) column chromatography, and chloroform-methanol (95: 5-85: 15) wash-out gets position D (3.2g), silica gel column chromatography, and chloroform-methanol (15: 1) wash-out gets required compound 60mg.
The white amorphous powder.C
25H
41NO
9。
ESIMS?m/z(％)：472[M+H]
+(100)；
1H?NMR(CDCl
3)δ：4.51(1H，d，J＝4.0Hz)，4.31(1H，d，J＝5.2Hz)，4.17(1H，d，J＝2.4Hz)，3.60(3H，s)，3.43(3H，s)，3.39(3H，s)，3.34(3H，s)，3.26(3H，s)；
13C?NMR(CDCl
3)δ：91.6(d)，81.7(d)，80.2(d)，79.9(d)，77.9(s)，78.2(d)，76.8(t)，76.0(s)，69.1(d)，66.9(d)，60.4(q)，59.0(q)，57.2(q)，54.1(q)，51.5(t)，50.3(s)，43.2(s)，42.3(d)，40.5(d)，40.5(d)，40.4(d)，36.4(t)，29.0(t)。
Embodiment 2: the preparation of the peaceful alkali of different Dare (XIII)
Silica gel column chromatography gained B position (2.1g) among the embodiment 1, silica gel column chromatography, chloroform-methanol (8: 2 → 7: 3) wash-out gets required compound 25mg.
The white amorphous powder.C
24H
39NO
7。
ESI-MS?m/z(％)：454[M+H]
+(100)；
1H?NMR(CDCl
3)δ：4.38(1H，d，J＝6.8Hz)，4.21(1H，d，J＝7.0Hz)，4.07(1H，d，J＝4.4Hz)，3.73(1H，d，J＝3.2Hz)，3.54(1H，d，J＝8.4Hz)，3.41(3H，s)，3.34(3H，s)，3.29(6H，s)，2.40(3H，s)；
13C?NMR(CDCl
3)δ：92.2(d)，84.5(t)，80.6(d)，79.4(d)，79.2(s)，76.3(d)，72.9(d)，63.6(d)，59.1(q)，58.9(t)，58.1(q)，57.5(q)，57.2(q)，50.6(s)，49.8(d)，49.3(d)，48.4(d)，45.3(d)，44.4(d)，41.6(q)，39.3(s)，31.7(t)，30.7(t)，29.8(t)。
Embodiment 3: the preparation of Bei Wuning (XIV)
Take by weighing Bei Wuting (beiwutine) 1.0g, add 20 milliliters of 5%NaOH methanol solutions, ambient temperature overnight, decompressing and extracting gets white solid thing 0.7g.Silica gel column chromatography, the chloroform-methanol that ammoniacal liquor is saturated (7: 3) wash-out gets required compound.
Yield: 92%.The white amorphous powder.C
24H
39N0
10。
ESI-MS?m/z(％)：502[M+H]
+(100)；
1H?NMR(CDCl
3)δ：4.45(1H，d，J＝6.0Hz)，4.26(1H，d，J＝5.2Hz)，3.16(1H，d，J＝6.4Hz)，3.82(1H，d，J＝8.4Hz)，3.57(3H，s)，3.35(3H，s)，3.29(3H，s)，3.27(3H，s)，2.37(3H，s)；
13C?NMR(CDCl
3)δ：92.6(d)，84.4(d)，83.2(d)，79.4(s)，78.5(d)，78.4(d)，78.1(s)，77.7(s)，75.8(t)，70.6(d)，61.5(q)，59.4(d)，59.1(q)，58.4(q)，56.8(s)，55.9(q)，51.1(t)，49.0(q)，49.0(t)，47.8(d)，44.4(s)，42.8(d)，42.4(d)，34.7(t)，34.7(t)。
Embodiment 4: the extraction separation of napelline (IV)
Take by weighing Xinjiang and produce the rhizome of Chinese monkshood (Aconitum carchaeli Debx.) rhizome powder 10kg, contain and infiltrate in the filter cylinder, with the diafiltration of 10～20 times of 0.3%HCl water liquid, after the alkalization of percolate strong aqua, use ethyl acetate extraction, the extraction liquid decompressing and extracting must the brown solid thing.
Silica gel column chromatography (ETHYLE ACETATE-sherwood oil-diethylamine, 60: 40: 2, w/w) separate, get the bullion napelline.Through 95% alcohol crystal, get required compound again.
Yield: 0.7%.Colourless needle crystal.C
34H
47NO
11。
ESI-MS?m/z(％)：646[M+H]
+(100)；
1H?NMR(CDCl
3)δ：8.03(1H，d，J＝7.2Hz)，7.58(1H，t，J＝7.2Hz)，7.46(1H，t，J＝7.2Hz)，4.87(1H，d，J＝5.2Hz)，4.47(1H，dd，J＝5.2，2.8Hz)，4.39(1H，d，J＝2.8Hz)，4.03(1H，d，J＝6.4Hz)，3.96(3H，s)，3.75(3H，s)，3.62(1H，d，J＝9.2Hz)，3.50(1H，d，J＝8.8Hz)，3.34(1H，d，J＝5.2Hz)，3.30(3H，s)，3.26(3H，s)，3.16(3H，s)，1.39(3H，s)，1.10(3H，d，J＝7.2Hz)；
13C?NMR(CDCl
3)δ：172.4(s)，166.0(s)，133.3(d)，129.6(s)，129.5(d)，128.6(d)，91.9(s)，89.9(d)，83.3(d)，82.3(d)，82.3(d)，78.8(d)，78.8(d)，76.6(t)，74.0(s)，71.3(d)，61.1(q)，61.0(d)，59.1(q)，57.9(q)，55.9(q)，49.9(s)，48.9(t)，46.9(d)，46.7(d)，44.6(d)，44.1(d)，43.0(s)，40.8(d)，35.7(t)，33.5(t)，21.4(q)，13.3(q)。
Embodiment 5: the preparation of the peaceful alkali of middle crow
(1) 3,13, the preparation of 15-triacetyl napelline (V)
Take by weighing 18.0 moles of napellines, add in the there-necked flask, other adds tosic acid 6g and 72 milliliters of diacetyl oxides, stirs, and room temperature reaction was handled by ordinary method after 72 hours, got the white solid thing.Through silica gel column chromatography (or acetone-ether crystallization), get required compound.
Yield: 95%.Colourless needle crystal.C
40H
53NO
14。
ESI-MS?m/z(％)：772[M+H]
+(100)；
1H?NMR(CDCl
3)δ：8.16(1H，d，J＝7.2Hz)，7.56(1H，t，J＝7.2Hz)，7.48(1H，t，J＝7.2Hz)，6.09(1H，d，J＝5.6Hz)，5.13(1H，d，J＝5.2Hz)，4.91(1H，dd，J＝8.4，5.2Hz)，4.11(1H，m)，4.05(1H，d，J＝5.6Hz)，3.75(1H，d，J＝8.8Hz)，3.68(1H，d，J＝5.2Hz)，3.64(1H，d，J＝5.2Hz)，3.40(3H，s)，3.24(3H，s)，3.19(6H，s)，3.11(1H，dd，J＝10.0，7.2Hz)，2.14(3H，s)，2.07(3H，s)，2.06(3H，s)，2.01(3H，s)，1.23(3H，s)，1.16(3H，t，J＝7.2Hz)；
13C?NMR(CDCl
3)δ：170.3(s)，170.3(s)，169.7(s)，168.8(s)，166.2(s)，133.3(d)，129.9(d)，129.5(s)，128.7(d)，88.7(d)，83.6(d)，81.5(d)，81.0(d)，78.8(d)，76.7(d)，71.3(t)，61.4(q)，60.4(d)，58.7(q)，58.6(q)，56.1(q)，49.8(s)，48.9(t)，47.0(t)，45.5(d)，45.1(d)，43.9(d)，42.1(s)，41.2(d)，35.9(t)，31.9(t)，21.2(q)，21.2(q)，21.1(q)，21.1(q)，13.6(q)。
(2) N-removes ethyl-3,13, the preparation of 15-triacetyl napelline (VI)
Take by weighing 17.0 mole 3,13,15-triacetyl napelline adds in the there-necked flask, and other adds 420 milliliters in Glacial acetic acid min. 99.5 and 54.5 moles of N-bromo-succinimides, stirs, and room temperature reaction got the white solid thing by the ordinary method processing reaction after 4 hours.Through silica gel column chromatography (sherwood oil-acetone, 2: 1), get required compound.
Yield: 75%.The white amorphous powder.C
38H
49NO
14。
ESI-MS?m/z(％)：744[M+H]
+(100)；
1H NMR (CDCl
3) δ: 7.48-8.18 (5H, m, Ar-H), 6.10 (1H, d, J=6.0Hz, H-15), 5.16 (1H; D, J=5.2Hz, H-14 β), 5.04 (1H, dd, J=10.0,5.6Hz; H-3), 4.07 (1H, d, J=6.8Hz, H-6), 3.86 (1H, d; J=5.6Hz, H-16), 3.75,2.97 (each 1H, ABq, J=8.8Hz, H
2-18), 3.21,3.23,3.30,3.40 (each 3H, s, OAc * 4), 2.06,2.07,2.18 (each 3H, s, OAc * 3), 1.25 (3H, s, OAc-8);
13C?NMR(CDCl
3)δ：170.5(s)，170.4(s)，169.9(s)，168.7(s)，166.1(s)，133.4(d)，129.9(d)，129.3(s)，128.7(d)，88.3(d)，88.3(s)，83.7(d)，80.8(s)，80.6(d)，78.7(d)，76.7(d)，72.9(t)，71.8(d)，61.4(d)，58.8(q)，58.5(q)，55.8(q)，55.7(q)，51.4(d)，49.9(s)，44.6(d)，42.9(d)，42.7(s)，41.3(d)，41.1(t)，34.6(t)，31.7(t)，21.3(q)，21.3(q)，21.1(q)，21.1(q)。
(3) 3,13, the preparation of black alkali (VII) in the 15-triacetyl
Take by weighing N-and remove ethyl-3,13,13 moles of 15-triacetyl napellines; Contain in the there-necked flask, other adds 13 moles of sodium hydrides and 300 milliliters of THFs, stirs 10 minutes under the argon gas stream; Add methyl iodide 15 mmoles again, stir room temperature reaction 3 hours; Drain solvent, by ordinary method handle the white solid thing.Through silica gel column chromatography (chloroform-methanol=95: 1), get required compound.
Yield: 90%.Colourless prism.C
39H
51NO
14。
ESI-MS?m/z(％)：758[M+H]
+(100)；
1H NMR (CDCl
3) δ: 7.49-8.17 (5H, m), 6.07 (1H, d, J=5.6Hz), 5.14 (1H, d, J=5.2Hz), 4.91 (1H, dd; J=12.8,6.0Hz), 4.05 (1H, d, J=6.8Hz), 3.85 (1H, d, J=5.2Hz), 3.40,3.27; 3.27,3.19 (each 3H, s), 3.76,2.95 (each 1H, ABq, J=8.8Hz), 2.66 (1H, ABq; J=11.2Hz), 2.45 (3H, s), 2.16,2.07,2.06 (each 3H, s), 1.24 (3H, s);
13C?NMR(CDCl
3)δ：170.3(s)，170.1(s)，169.5(s)，168.7(s)，166.2(s)，133.3(d)，130.0(d)，129.6(s)，128.6(d)，88.8(d)，88.6(s)，83.5(d)，81.6(d)，81.1(s)，78.9(d)，76.7(d)，71.3(d)，71.2(t)，62.0(d)，61.2(q)，58.7(q)，58.6(q)，56.3(q)，49.9(s)，49.6(t)，45.0(d)，44.3(d)，43.9(d)，42.6(q)，42.4(s)，41.3(d)，35.7(t)，31.9(t)，21.2(q)，21.1(q)，21.1(q)，21.1(q)。
(4) preparation of the peaceful alkali (II) of crow in
Take by weighing 3,13, black alkali is 10.0 moles in the 15-triacetyl, contains in the there-necked flask, and other adds 250 milliliters of 5% sodium hydroxide methanol solutions, and 60 ℃ were stirred 30 minutes, drained solvent, handled by ordinary method, got the white solid thing.Through silica gel column chromatography (chloroform-methanol, 95: 5), get required compound.
Yield: 95%.The white amorphous powder.C
24H
39NO
9。
Spectrum (IR,
1HNMR,
13CNMR, ESI-MS) data see before and state.
Embodiment 6: the preparation of the peaceful alkali of inferior crow
(1) preparation of deoxyaconitine (VIII)
Take by weighing 2.0 moles of napellines, contain in the there-necked flask, other adds 100 milliliters of THFs and 7 milliliters of sulfur oxychlorides, stirs, and room temperature reaction was handled by ordinary method after 2 hours, the tawny solids.It is contained in the there-necked flask, add each 80 milliliters of ETHYLE ACETATE and 95% ethanol, other adds 2 milliliters in Glacial acetic acid min. 99.5, and palladium carbon 1 gram stirs and feeds hydrogen (about 0.1kg/cm down
2), react after 72 hours, get the white solid thing by ordinary method.Through silica gel column chromatography (chloroform-methanol, 95: 5), get required compound.
Yield: 90%.Colourless needle crystal.C
34H
47NO
10。
ESI-MS?m/z(％)：630[M+H](100)；
1H?NMR(CDCl
3)δ：8.02(1H，d，J＝7.2Hz)，7.57(1H，t，J＝7.2Hz)，7.45(1H，t，J＝7.2Hz)，4.87(1H，d，J＝4.8Hz)，4.45(1H，dd，J＝5.2，2.8Hz)，4.37(1H，d，J＝2.8Hz)，3.97(1H，d，J＝6.0Hz)，3.89(1H，s)，3.73(3H，s)，3.63(1H，d，J＝8.4Hz)，3.28(3H，s)，3.26(3H，s)，3.15(3H，s)，3.10(1H，d，J＝8.4Hz)，1.36(3H，s)，1.06(3H，t，J＝7.2Hz)；
13C?NMR(CDCl
3)δ：172.4(s)，166.1(s)，133.2(d)，129.7(d)，129.6(s)，128.5(d)，92.0(s)，90.0(d)，84.9(d)，83.1(d)，80.2(t)，78.9(d)，78.8(d)，74.0(s)，61.4(d)，61.0(q)，58.6(q)，57.9(q)，56.3(q)，53.0(t)，49.8(s)，49.2(t)，49.0(d)，45.0(d)，44.5(d)，40.9(d)，39.0(s)，36.6(t)，35.2(t)，26.3(t)，21.4(q)，13.4(q)。
The preparation of (2) 13,15-diacetyl deoxyaconitines (IX)
Take by weighing 3.0 moles of Shengs of deoxyaconitine and go in the there-necked flask, other adds tosic acid 2 gram and 20 milliliters of diacetyl oxides, stirs, and 45 ℃ of reactions were handled by ordinary method after 48 hours, got the white solid thing.Through acetone-ether crystallization, get required compound.
Yield: 90%.The white amorphous powder.C
38H
51NO
12。
ESI-MS?m/z(％)：714[M+H]
+(100)；
1H?NMR(CDCl
3)δ：8.16(1H，d，J＝7.2Hz)，7.56(1H，t，J＝7.2Hz)，7.48(1H，t，J＝7.2Hz)，6.07(1H，d，J＝5.6Hz)，5.14(1H，d，J＝5.2Hz)，3.94(1H，d，J＝6.4Hz)，3.84(1H，d，J＝5.6Hz)，3.40(3H，s)，3.25(3H，s)，3.22(3H，s)，3.15(3H，s)，2.13(3H，s)，2.04(3H，s)，1.25(3H，s)，1.13(3H，t，J＝7.2Hz)；
13C?NMR(CDCl
3)δ：170.3(s)，169.8(s)，168.8(s)，166.3(s)，133.3(d)，129.9(d)，129.5(s)，128.7(d)，88.9(s)，88.7(d)，84.7(d)，83.4(d)，81.1(s)，80.0(t)，78.9(d)，76.9(d)，61.4(q)，61.2(d)，59.6(q)，58.4(q)，53.0(q)，50.1(s)，49.1(t)，49.1(t)，48.7(d)，45.1(d)，43.9(d)，41.7(d)，38.8(s)，36.2(t)，35.2(t)，26.3(t)，21.3(q)，21.2(q)，21.1(q)，13.6(q)。
(3) N-removes ethyl-13, the preparation of 15-diacetyl deoxyaconitine (X)
Take by weighing 13,5.0 moles of 15-diacetyl deoxyaconitines are contained in the there-necked flask, and other adds 90 milliliters in Glacial acetic acid min. 99.5 and 15 moles of N-bromo-succinimides, and the stirring at room reaction was handled by ordinary method after 3 hours, the white solid thing.Through silica gel column chromatography (the saturated chloroform-methanol of ammoniacal liquor, 8: 2), get required compound.
Yield: 75%.The white amorphous powder.C
32H
43NO
10。
ESI-MS?m/z(％)：602[M+H]
+(100)；
1H?NMR(CDCl
3)δ：8.17(1H，d，J＝7.2Hz)，7.57(1H，t，J＝7.2Hz)，7.49(1H，t，J＝7.2Hz)，6.16(1H，d，J＝6.5Hz)，5.17(1H，d，J＝4.8Hz)，3.93(1H，dd，J＝9.6，6.4Hz)，3.41(3H，s)，3.29(3H，s)，3.27(3H，s)，3.18(3H，s)，2.21(3H，s)，2.08(3H，s)，1.26(3H，s)；
13C?NMR(CDCl
3)δ：170.6(s)，170.5(s)，168.7(s)，166.1(s)，133.4(d)，130.0(d)，129.3(s)，128.7(d)，88.7(s)，87.9(d)，83.7(d)，82.5(d)，80.7(s)，79.8(t)，78.6(d)，77.0(d)，61.5(d)，59.1(q)，58.4(q)，56.8(q)，55.5(q)，50.1(s)，49.2(d)，49.0(t)，43.4(d)，42.7(d)，41.1(d)，39.0(s)，35.4(t)，29.7(t)，29.6(t)，24.0(t)。
The preparation of (4) 13,15-diacetyl time black alkali (XI)
Take by weighing N-demethyl-13,3.8 moles of 15-diacetyl deoxyaconitines, Sheng is gone in the there-necked flask, and other adds 3.8 moles of sodium hydrides, and 4 moles of 80 milliliters of THFs and methyl iodide stir, and room temperature reaction was handled by ordinary method after 3 hours, got the white solid thing.Through silica gel column chromatography (chloroform-methanol, 95: 5), get required compound.
Yield: 80%.The white amorphous powder.C
33H
49NO
12。
ESI-MS?m/z(％)：698[M+H]
+(100)；
1H?NMR(CDCl
3)δ：8.17(1H，d，J＝7.2Hz)，7.52(1H，t，J＝7.2Hz)，7.48(1H，t，J＝7.2Hz)，6.07(1H，d，J＝5.6Hz)，5.13(1H，d，J＝5.2Hz)，3.40(3H，s)，3.27(6H，s)，3.15(3H，s)，2.42(3H，s)，2.13(3H，s)，2.03(3H，s)，1.22(3H，s)；
13C?NMR(CDCl
3)δ：170.3(s)，169.7(s)，168.8(s)，166.2(s)，133.3(d)，129.9(d)，129.5(s)，128.6(d)，88.7(s)，88.7(d)，84.6(d)，83.3(d)，81.1(s)，79.8(t)，78.9(d)，76.8(d)，62.3(d)，61.3(q)，58.9(q)，58.3(q)，56.3(q)，55.7(t)，50.1(s)，47.6(q)，44.0(d)，43.8(d)，42.8(d)，41.1(d)，39.0(s)，35.9(t)，34.8(t)，26.4(t)，21.3(q)，21.2(q)，21.1(q)。
(5) preparation of the peaceful alkali of inferior crow (III)
Take by weighing 13,5.0 moles in 15-diacetyl time black alkali is contained in the there-necked flask, and other adds 40 milliliters of 5% sodium hydroxide methanol solutions, and 60 ℃ of stirring reactions were handled by ordinary method after 30 minutes, got the white solid thing.Separate at 95: 5 through the saturated chloroform-methanol of silica gel column chromatography ammoniacal liquor, get required compound.
Yield: 92%.The white amorphous powder.C
24H
39NO
8。
Spectrum (ESI-MS,
1H NMR,
13C NMR HR-MS) data are seen before.
Embodiment 7:N-goes the preparation of the ethyl time peaceful alkali (XV) of crow
Take by weighing N-demethyl-13 among the embodiment 5,15-diacetyl time black alkali 0.5 gram is contained in the there-necked flask, adds 15 milliliters of 5%NaOH methanol solutions, room temperature, and stirred overnight is drained, and gets the white solid thing.Silica gel column chromatography (chloroform-methanol, 7: 3) gets required compound.
Yield: 92%.The white amorphous powder.C
23H
37NO
8。
Spectrum (
1H NMR,
13C NMR, ESI-MS, HR-MS) data see before.
The representational pharmaceutical dosage form of elucidated hereinafter is applicable to that compound shown in the general formula (I) and the peaceful alkali of middle crow and time peaceful alkali of crow or its pharmaceutical salts (below be designated as " compounds X ") take when treatment.
Claims (3)
1.C
19The peaceful alkali (II) of crow and time peaceful alkali (III) of crow or the purposes of their pharmaceutical salts in preparation cardiotonic drug and anti-heart failure agent in-the diterpene alkaloid,
2. be used for preparing the preparation method of the peaceful alkali of crow of cardiotonic drug and anti-heart failure agent, may further comprise the steps:
1) under the Catalyzed by p-Toluenesulfonic Acid, napelline (aconitine) (IV) with Acetyl Chloride 98Min. or aceticanhydride room temperature reaction 6-12 hour, after well-established law is handled, solids; 95% alcohol crystal gets compound 3,13; 15-triacetyl napelline (3,13,15-triacetylaconitine) (V);
2) 3,13,15-triacetyl napelline mixes with 3 normal N-bromo-succinimides and 20-40 times of Glacial acetic acid min. 99.5; Room temperature reaction 2-6 hour, concentrating under reduced pressure got the white solid thing, and silica gel column chromatography separates; With sherwood oil, acetone, chloroform, methylene dichloride, methyl alcohol or their mixed solvent is elutriant, gets N-and removes ethyl-3,13; 15-triacetyl napelline (N-deethyl-3,13,15-triacetylaconitine) (VI);
3) N-removes ethyl-3,13, and 15-triacetyl napelline mixes with normal methyl iodide, and THF is a solvent; Room temperature reaction 2-4 hour, decompressing and extracting got the white solid thing, and silica gel column chromatography separates; With sherwood oil, ETHYLE ACETATE, chloroform, methyl alcohol or their mixed solvent is elutriant, gets 3,13; Black alkali in the 15-triacetyl (3,13,15-triacetylmesaconitine) (VII);
4) 3,13, black alkali is used 5% sodium hydroxide hydrolysis, ambient temperature overnight in the 15-triacetyl; Decompressing and extracting gets the white solid thing, and silica gel column chromatography separates; With sherwood oil, chloroform, methylene dichloride, methyl alcohol or their mixing solutions is eluent, gets the peaceful alkali (II) of crow in the compound
3. be used to prepare time black rather preparation method of alkali of cardiotonic drug and anti-heart failure agent, may further comprise the steps:
1) with the THF is solvent, napelline is mixed with normal thionyl chloride, ambient temperature overnight reaction 1-3 hour; Decompressing and extracting, the white solid thing, add ETHYLE ACETATE-95% ethanol-Glacial acetic acid min. 99.5 mixed solution again after; Under the catalysis of palladium carbon, normal pressure hydrogenation 1-3 days, decompressing and extracting; Get the white solid thing,, get deoxyaconitine (deoxyaconitine) (VIII) through 95% alcohol crystal;
2) under the Catalyzed by p-Toluenesulfonic Acid, deoxyaconitine mixes with 1～2 normal Acetyl Chloride 98Min. or aceticanhydride, stirs; Room temperature reaction spends the night, and decompressing and extracting gets the white solid thing; Use 95% alcohol crystal again; Get compound 13,15-diacetyl deoxyaconitine) (13,15-diacetyldeoxyaconitine) (IX);
3) 13,15-diacetyl deoxyaconitine mixes with 2-4 equivalent N-bromo-succinimide and 20-40 times of Glacial acetic acid min. 99.5, room temperature reaction 2-6 hour; Concentrating under reduced pressure gets the white solid thing, and silica gel column chromatography separates; With sherwood oil, acetone, chloroform, methyl alcohol or their mixed solvent is eluent; Compound N-go ethyl-13,15-diacetyl deoxyaconitine (N-deethyl-13,15-diacetyl-deoxyaconitine) (X);
4) N-removes ethyl-13, and 15-diacetyl deoxyaconitine mixes with the equivalent methyl iodide, and THF is a solvent; Room temperature reaction 2-4 hour, decompressing and extracting got the white solid thing; Silica gel column chromatography separates, and is elutriant with sherwood oil, ETHYLE ACETATE, chloroform, acetone, methyl alcohol or their mixed solvent, gets compound 13; 15-diacetyl time black alkali (13,15-diacetylhypaconitine) (XI);
5) 13, and 15-diacetyl time black alkali (13,15-diacetylhypaconitine) add 5% sodium hydroxide solution; Ambient temperature overnight, well-established law is handled, and gets the white solid thing; Silica gel column chromatography separates; With chloroform, ETHYLE ACETATE, methylene dichloride, acetone, methyl alcohol or their mixed solvent is elutriant, gets the peaceful alkali (III) of compound time crow
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| CN102977020A (en) * | 2012-12-05 | 2013-03-20 | 四川大学 | Aconitine alkaloids and preparation method thereof, and pharmaceutical composition using compounds as cardiotonic and anti-heart-failure active component and application thereof |
| CN108456168B (en) * | 2017-02-21 | 2021-11-05 | 西南交通大学 | Class C19Diterpene alkaloid and application thereof |
| CN110066247B (en) * | 2018-01-24 | 2021-01-01 | 好医生药业集团有限公司 | Zhongwuning crystal form and preparation method thereof |
| CN108912049B (en) * | 2018-04-25 | 2020-08-11 | 西安医学院 | Diterpene alkaloid compound extracted from aconitum sinomontanum nakai and preparation method and application thereof |
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