CN105232891B - The preparation method of trilliaceae extract and wherein saponins compound and its preparing the application in anti-ischemic heart medicine - Google Patents
The preparation method of trilliaceae extract and wherein saponins compound and its preparing the application in anti-ischemic heart medicine Download PDFInfo
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- CN105232891B CN105232891B CN201510697362.5A CN201510697362A CN105232891B CN 105232891 B CN105232891 B CN 105232891B CN 201510697362 A CN201510697362 A CN 201510697362A CN 105232891 B CN105232891 B CN 105232891B
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Abstract
The present invention relates to disclose trilliaceae total steroidal saponin and the wherein preparation method of steroid saponin compound and its preparing the application in anti-ischemic heart medicine.Prepared trilliaceae total steroidal saponin itself is the 60-85% alcohol elution of macroporous absorbent resin on trilliaceae total extract and its total steroidal saponin content is greater than 90%, mainly includes bisnosaponin compound content 66-75%, Trillin kind compound content 3-7%, Dioscin kind compound content 5-8%.Prepared trilliaceae steroid saponin compound itself is one of pennogenin -3-O- α-L- rhamnopyranosyl (1 → 2)-β-D-Glucose glycosides, pennogenin -3-O- α-L- rhamnopyranosyl (1 → 4)-[α-L- rhamnopyranosyl (1 → 2)]-β-D-Glucose glycosides and pennogenin -3-O- α-L- rhamnopyranosyl (1 → 4)-α-L- rhamnopyranosyl (1 → 4)-[α-L- rhamnopyranosyl (1 → 2)]-β-D-Glucose glycosides or a variety of.Above-mentioned trilliaceae total steroidal saponin and wherein steroid saponin compound are used to prepare anti-ischemic heart medicine, and any one for the ischemic heart disease that there is the coronary heart diseases and angina pectoris and myocardial infarction treating and caused and occurred by myocardial ischemia to be formed.
Description
Technical field
The present invention relates to trilliaceae extract and wherein the preparation method of saponins compound and its preparing anti-ischemic
Application in cardiotropic formulation, belongs to technical field of traditional Chinese medicine preparation.
Background technique
Trilliaceae is Liliaceae Trillium plant trilliaceae Trillium tschonoskii Maxim., the popular name crown
Rhizoma Trillii Tschonoskii, Rhizoma Trillii Tschonoskii, lion seven etc., are traditional rare Chinese medicines, there is the effect of promoting longevity.It cures mainly and has a dizzy spell, has a sleepless night, falling
Beat the diseases such as damage, traumatic hemorrhage, neurasthenia, high blood pressure, postconcussion syndrome, be the famous Folk medicine of Tujia Nationality in Enshi it
One.Trilliaceae, which has stronger anti-inflammatory, immunological regulation, improvement, to be shown to the pharmacology activity research of Trillium plant both at home and abroad at present
The effects of learning and memory function and anti-aging.Inventor causes myocardial ischemia in rats experimental model using coronary ligation, from the heart
Electrograph, myocardial infarct size, serum creatine kinase (CK) and vigor of serum lactic dehydrogenase (SLDH) (LDH) etc. are studied and confirm to prolong
Age 85% ethanol elution object of grass has significant preventive and therapeutic effect to myocardial ischemia.
Chemical research shows 85% alcohol elution of active component based on steroid saponin constituents, and steroid saponin is total
Content be greater than 90%, wherein predominantly pennogenin -3-O- α-L- rhamnopyranosyl (1 → 2)-β-D-Glucose glycosides, partially
Promise sapogenin -3-O- α-L- rhamnopyranosyl (1 → 4)-[α-L- rhamnopyranosyl (1 → 2)]-β-D-Glucose glycosides and partially
Promise sapogenin -3-O- α-L- rhamnopyranosyl (1 → 4)-α-L- rhamnopyranosyl (1 → 4)-[α-L- rhamnopyranosyl
(1 → 2)]-β-D-Glucose glycosides, above-mentioned three kinds of saponin(es are both the index and characteristic chemical constituent and effective of trilliaceae medicinal material
The index and characteristic chemical constituent of 85% alcohol elution of position.Further pharmacological research confirms pennogenin in active component
Member -3-O- α-L- rhamnopyranosyl (1 → 2)-β-D-Glucose glycosides, pennogenin -3-O- α-L- rhamnopyranosyl (1
→ 4)-[α-L- rhamnopyranosyl (1 → 2)]-β-D-Glucose glycosides and pennogenin -3-O- α-L- rhamnopyranosyl (1
→ 4)-α-L- rhamnopyranosyl (1 → 4)-[α-L- rhamnopyranosyl (1 → 2)]-β-D-Glucose glycosides is that trilliaceae is anti-scarce
The effective component of hemorrhagic cardiotropic formulation.The studies above is not yet reported that we pass through the inside and outside experimental study of system, first
It was found that it, which fights ischemic heart disease, has good therapeutic effect.
Summary of the invention
It is an object of the present invention to provide a kind of trilliaceae extract and wherein the preparation method of saponins compound and its making
Application in standby anti-ischemic heart medicine.
Present invention employs following technical proposals.
The preparation method of trilliaceae extract: by trilliaceae pulverizing medicinal materials at coarse powder, 0.5-2 is impregnated with 0-100% ethyl alcohol
Hour, 4-12 times is measured solvent heating and refluxing extraction 2-4 times, and each 1-3 hours, filtering, merging filtrate.Above-mentioned filtrate and 45-80
It DEG C is concentrated under reduced pressure into 0.5-1.5g raw medicinal herbs/ml, 0-4 DEG C of refrigerated overnight analyses glue, filtering, and filtrate is recycled with extracting n-butyl alcohol again
It obtains medicinal extract and dilutes the directly upper macroporous absorbent resin AB-8 type of upper macroporous absorbent resin or filtrate or D-101 type or HPD-400 or each
Kind model macroporous adsorbing resin for purification, is successively eluted with ethanol water by 0-50%, then afforded respectively with ethanol water by 50-95%
Position, the main elution position for collecting ethanol water 60-85% are eluted, solvent is recovered under reduced pressure at 45-80 DEG C to second for each position that elutes
The appropriate concentration of alcohol is dried with freeze-drying or spray drying, collect freeze-dried powder or spray dried powder to get.
Pennogenin -3-O- α-L- rhamnopyranosyl (1 → 2)-β-D-Glucose in Trillin class compound
Glycosides (pennogenin-3-O- α-L-rhamnopyranosyl- (1 → 2)-β-D-glucopyranoside), pennogenin-
3-O- α-L- rhamnopyranosyl (1 → 4)-[α-L- rhamnopyranosyl (1 → 2)]-β-D-Glucose glycosides (pennogenin-
3-O-α-L-rhamnopyranosyl-(1→4)-[a-L-rhamnopyranosyl-(1→2)]-β-D-
Glucopyranoside) and pennogenin -3-O- α-L- rhamnopyranosyl (1 → 4)-α-L- rhamnopyranosyl (1 →
4)-[α-L- rhamnopyranosyl (1 → 2)]-β-D-Glucose glycosides (pennogenin-3-O- α-L-rhamnopyranosyl-
(1→4)-α-L-rhamnopyranosyl-(1→4)-[α-L-rhamnopyranosyl-(1→2)]-β-D-
Glucopyranoside by silicagel column on gained trilliaceae extract, chloroform: first successively preparation method): is used after loading
Alcohol or methylene chloride: methanol or chloroform: methanol: water or methylene chloride: methanol: water gradient elution, with above-mentioned trilliaceae soap
Pennogenin -3-O- α-L- rhamnopyranosyl (1 → 2)-β-D-Glucose glycosides, pennogenin -3- in glycosides compound
O- α-L- rhamnopyranosyl (1 → 4)-[α-L- rhamnopyranosyl (1 → 2)]-β-D-Glucose glycosides and pennogenin -3-
O- α-L- rhamnopyranosyl (1 → 4)-α-L- rhamnopyranosyl (1 → 4)-[α-L- rhamnopyranosyl (1 → 2)]-β-D-
3 kinds of saponin(es of glucoside are detected as reference substance TLC or HPLC, merge corresponding fraction.Above-mentioned fraction is dense in 45-80 DEG C of decompression
Contracting is evaporated, and is suspended or is dissolved with appropriate distilled water, upper reverse phase silica gel ODS-C18 column chromatography or SephadexLH-20 column chromatography or
Preparative high-performance liquid chromatographic technology isolates and purifies, and is successively eluted with ethanol water by 0-50%, then is eluted with ethanol water 50-95%,
Eluent freeze-drying or spray drying are dried, and freeze-dried powder or spray dried powder are collected, with 10-95% alcohol crystal
And recrystallize to get.
The trilliaceae extract of preceding method preparation and wherein saponins compound, effective component are pennogenin-
3-O- α-L- rhamnopyranosyl (1 → 2)-β-D-Glucose glycosides, pennogenin -3-O- α-L- rhamnopyranosyl (1 →
4)-[α-L- rhamnopyranosyl (1 → 2)]-β-D-Glucose glycosides and pennogenin -3-O- α-L- rhamnopyranosyl (1 →
4) one of-α-L- rhamnopyranosyl (1 → 4)-[α-L- rhamnopyranosyl (1 → 2)]-β-D-Glucose glycosides or more
The hydroxyl or 17 hydroxyls of 21st and the 27th conversion methylol of kind or the sapogenin in its substance or all glycosyls
Alcoholic extract hydroxyl group is replaced by any one in acetyl group, fatty alkenyl group, aromatic substituents, amide groups, or hydrolyze
The substance that pennogenin -3-O- α-L- rhamnopyranosyl (1 → 2)-β-D-Glucose glycosides can be generated afterwards, can be used for preparing anti-
Ischemic heart medicine, the drug include oral solution, capsule, tablet, effervescent tablet, powder-injection, liquid drugs injection or injection or
Various dosage formulations, the drug further include various folk prescriptions and compound preparation.
Related ischemic heart disease is the coronary heart diseases and angina pectoris for being caused and being occurred by myocardial ischemia and myocardial infarction etc.
Any one of ischemic heart disease.
The invention has the advantages that providing a kind of trilliaceae extract and the wherein extraction of saponins compound and preparation side
Method, effective component is mainly pennogenin -3-O- α-L- rhamnopyranosyl (1 → 2)-β-D-Glucose in obtained material
Glycosides, pennogenin -3-O- α-L- rhamnopyranosyl (1 → 4)-[α-L- rhamnopyranosyl (1 → 2)]-β-D-Glucose
Glycosides and pennogenin -3-O- α-L- rhamnopyranosyl (1 → 4)-α-L- rhamnopyranosyl (1 → 4)-[α-L- pyrans mouse
Lee's glycosyl (1 → 2)] one of-β-D-Glucose glycosides or a variety of, obtained material can be used for preparing anti-ischemic heart medicine
Oral solution, capsule, tablet, effervescent tablet, powder-injection, liquid drugs injection or injection or various dosage formulations bulk pharmaceutical chemicals.
A kind of trilliaceae extract and wherein pennogenin -3-O- α-L- rhamnopyranosyl (1 in saponins compound
→ 2)-β-D-Glucose glycosides, pennogenin -3-O- α-L- rhamnopyranosyl (1 → 4)-[α-L- rhamnopyranosyl (1 →
2)]-β-D-Glucose glycosides and pennogenin -3-O- α-L- rhamnopyranosyl (1 → 4)-α-L- rhamnopyranosyl (1 →
4)-[α-L- rhamnopyranosyl (1 → 2)]-β-D-Glucose glycosides is in vivo and in vitro to the coronary disease for being caused by myocardial ischemia and being occurred
The ischemic heart diseases such as disease, angina pectoris and myocardial infarction have good therapeutic effect and stronger bioactivity.
A kind of trilliaceae extract and wherein pennogenin -3-O- α-L- rhamnopyranosyl (1 in saponins compound
→ 2)-β-D-Glucose glycosides, pennogenin -3-O- α-L- rhamnopyranosyl (1 → 4)-[α-L- rhamnopyranosyl (1 →
2)]-β-D-Glucose glycosides and pennogenin -3-O- α-L- rhamnopyranosyl (1 → 4)-α-L- rhamnopyranosyl (1 →
4)-[α-L- rhamnopyranosyl (1 → 2)] preparation method of-β-D-Glucose glycosides mainly by extracting from natural drug, is
Pure natural component and monomer have good biocompatibility.
Detailed description of the invention
Fig. 1 is the molecular structural formula of pennogenin -3-O- α-L- rhamnopyranosyl (1 → 2)-β-D-Glucose glycosides；
Fig. 2 is pennogenin -3-O- α-L- rhamnopyranosyl (1 → 4)-[α-L- rhamnopyranosyl (1 → 2)] -
β-D-Glucose glycosides molecular structural formula；
Fig. 3 is pennogenin -3-O- α-L- rhamnopyranosyl (1 → 4)-α-L- rhamnopyranosyl (1 → 4)-[α -
L- rhamnopyranosyl (1 → 2)]-β-D-Glucose glycosides molecular structural formula.
Specific embodiment
The present invention will be further described below with reference to examples:
Embodiment 1
A kind of trilliaceae extract and wherein pennogenin -3-O- α-L- rhamnopyranosyl (1 → 2)-β-D- grape
Glucosides, pennogenin -3-O- α-L- rhamnopyranosyl (1 → 4)-[α-L- rhamnopyranosyl (1 → 2)]-β-D- grape
Glucosides and pennogenin -3-O- α-L- rhamnopyranosyl (1 → 4)-α-L- rhamnopyranosyl (1 → 4)-[α-L- pyrans
Rhamnopyranosyl (1 → 2)]-β-D-Glucose glycosides preparation method: by trilliaceae pulverizing medicinal materials at coarse powder, impregnate 1 with 70% ethyl alcohol
Hour, 10 times amount solvent heating and refluxing extraction 3 times, 2 hours every time, filter, merging filtrate.Above-mentioned filtrate and 65 DEG C of reduced pressures
To 1.0g raw medicinal herbs/ml, 0-4 DEG C of refrigerated overnight analyses glue, filtering, and filtrate is extracted with organic solvents such as n-butanols again, recycles organic
Solvent obtains medicinal extract and water is added to be suspended, upper AB-8 model macroporous adsorbing resin for purification, successively with water, 30% ethyl alcohol, 60% ethyl alcohol and
85% ethanol elution obtains each elution position, is dried with freeze-drying, collect freeze-dried powder to get.Wherein 85% wash
Based on steroid saponin constituents, steroid saponin total content is greater than 95% to get trilliaceae extract at de- position.Prolong age for above-mentioned
Careless extract both the elution position of 85% ethyl alcohol medicinal extract be added equivalent 100-200 mesh silica gel mix thoroughly, 50 DEG C decompression volatilize it is molten
Agent is ground well, dry method loading, upper normal pressure silica gel column (200-300 mesh, silica gel amount are 30 times of medicinal extract), successively uses trichlorine after loading
Methane: methanol: water gradient elution mainly collects chloroform: methanol: the fraction of water 65: 35: 1, with above-mentioned Trillin
Pennogenin -3-O- α-L- rhamnopyranosyl (1 → 2)-β-D-Glucose glycosides, pennogenin -3-O- α-in substance
L- rhamnopyranosyl (1 → 4)-[α-L- rhamnopyranosyl (1 → 2)]-β-D-Glucose glycosides and pennogenin -3-O- α -
L- rhamnopyranosyl (1 → 4)-α-L- rhamnopyranosyl (1 → 4)-[α-L- rhamnopyranosyl (1 → 2)]-β-D- grape
3 kinds of saponin(es of glucosides are detected as reference substance TLC or HPLC, merge corresponding fraction.Above-mentioned fraction is evaporated in 65 DEG C of reduced pressures,
It is suspended with appropriate distilled water, upper reverse phase silica gel ODS-C18 column chromatography is successively eluted with ethanol water by 0-50%, then use ethanol water
50-95% elution, main 60%-80% eluent of collecting are dried with freeze-drying, collect freeze-dried powder, use 10-95%
Alcohol crystal and recrystallize to get.Pennogenin -3-O- α-L- pyrrole in Trillin substance is prepared using the above method
Mutter rhamnopyranosyl (1 → 2)-β-D-Glucose glycosides yield be 1.5-2.6g/kg, through HPLC detection purity be 99.0%；Inclined promise
The yield of sapogenin -3-O- α-L- rhamnopyranosyl (1 → 4)-[α-L- rhamnopyranosyl (1 → 2)]-β-D-Glucose glycosides
It is 98.6% through HPLC detection purity for 0.8-1.9g/kg；Pennogenin -3-O- α-L- rhamnopyranosyl (1 → 4)-α -
What the yield of L- rhamnopyranosyl (1 → 4)-[α-L- rhamnopyranosyl (1 → 2)]-β-D-Glucose glycosides respectively was
Yield is 0.3-0.9g/kg, is 98.2% through HPLC detection purity；Chromatographic condition UItimate RXB-C18 chromatographic column (4.6mm
× 150mm, 3 μm), mobile phase is acetonitrile-water (10%-100% gradient elution), Detection wavelength 203nm, flow velocity 1.0ml/
Min, 30 DEG C of column temperature, 20 μ l of sample volume.
Embodiment 2
A kind of trilliaceae extract and wherein pennogenin -3-O- α-L- rhamnopyranosyl (1 → 2)-β-D- grape
Glucosides, pennogenin -3-O- α-L- rhamnopyranosyl (1 → 4)-[α-L- rhamnopyranosyl (1 → 2)]-β-D- grape
Glucosides and pennogenin -3-O- α-L- rhamnopyranosyl (1 → 4)-α-L- rhamnopyranosyl (1 → 4)-[α-L- pyrans
Rhamnopyranosyl (1 → 2)]-β-D-Glucose glycosides preparation method: by trilliaceae pulverizing medicinal materials at coarse powder, impregnated with 60% ethyl alcohol
2.0 hours, 12 times amount solvent heating and refluxing extraction 3 times, 2 hours every time, filtering, merging filtrate.Above-mentioned filtrate and 70 DEG C of decompressions
It is concentrated into 1g raw medicinal herbs/ml, 0-4 DEG C of refrigerated overnight analyses glue, filters, D101 model macroporous adsorbing resin for purification on filtrate, successively
With water, 30% ethyl alcohol, 60% ethyl alcohol and 85% ethanol elution, each elution position is obtained, is dried with spray drying process, collected
Spray dried powder to get.Wherein based on steroid saponin constituents, steroid saponin total content is greater than 92% at 85% elution position,
Up to trilliaceae extract.The medicinal extract at the elution position of above-mentioned trilliaceae extract both 85% ethyl alcohol is added to the 100- of equivalent
200 mesh silica gel are mixed thoroughly, and 50 DEG C of decompressions volatilize solvent, grind well, dry method loading, and (200-300 mesh, silica gel amount are upper normal pressure silica gel column
25 times of medicinal extract), chloroform is successively used after loading: methanol: water gradient elution mainly collects chloroform: methanol: water 60
: 30: 1 elution fraction, with pennogenin -3-O- α-L- rhamnopyranosyl in above-mentioned Trillin substance (1 →
2)-β-D-Glucose glycosides, pennogenin -3-O- α-L- rhamnopyranosyl (1 → 4)-[α-L- rhamnopyranosyl (1 →
2)]-β-D-Glucose glycosides and pennogenin -3-O- α-L- rhamnopyranosyl (1 → 4)-α-L- rhamnopyranosyl (1 →
4)-[α-L- rhamnopyranosyl (1 → 2)] 3 kinds of saponin(es of-β-D-Glucose glycosides are detected as reference substance TLC or HPLC, are merged
Corresponding fraction.Above-mentioned fraction is evaporated in 65 DEG C of reduced pressures, is suspended with appropriate distilled water, upper SephadexLH-20 column chromatography, according to
It is secondary to be eluted with ethanol water by 0-50%, then eluted with ethanol water 50-95%, eluent is dried with spray drying, collects spray
Mist xeraphium, with 10-95% alcohol crystal and recrystallize to get.It is prepared in Trillin substance partially using the above method
The yield of promise sapogenin -3-O- α-L- rhamnopyranosyl (1 → 2)-β-D-Glucose glycosides is 1.5-2.6g/kg, is examined through HPLC
Surveying purity is 98.8%；Pennogenin -3-O- α-L- rhamnopyranosyl (1 → 4)-[α-L- rhamnopyranosyl (1 →
2)] yield of-β-D-Glucose glycosides is 0.8-1.9g/kg, is 98.5% through HPLC detection purity；Pennogenin -3-O- α -
L- rhamnopyranosyl (1 → 4)-α-L- rhamnopyranosyl (1 → 4)-[α-L- rhamnopyranosyl (1 → 2)]-β-D- grape
The yield that the yield of glucosides respectively is is 0.3-0.9g/kg, is 97.8% through HPLC detection purity；Chromatographic condition
UItimate RXB-C18 chromatographic column (4.6mm × 150mm, 3 μm), mobile phase are acetonitrile-water (10%-100% gradient elution),
Detection wavelength is 203nm, flow velocity 1.0ml/min, 30 DEG C of column temperature, 20 μ l of sample volume.
Embodiment 3
A kind of trilliaceae extract and wherein pennogenin -3-O- α-L- rhamnopyranosyl (1 → 2)-β-D- grape
Glucosides, pennogenin -3-O- α-L- rhamnopyranosyl (1 → 4)-[α-L- rhamnopyranosyl (1 → 2)]-β-D- grape
Glucosides and pennogenin -3-O- α-L- rhamnopyranosyl (1 → 4)-α-L- rhamnopyranosyl (1 → 4)-[α-L- pyrans
Rhamnopyranosyl (1 → 2)]-β-D-Glucose glycosides preparation method: by trilliaceae pulverizing medicinal materials at coarse powder, impregnated with 50% ethyl alcohol
1.0 hours, 8 times amount solvent heating and refluxing extraction 3 times, 2 hours every time, filtering, merging filtrate.Above-mentioned filtrate and 70 DEG C of decompressions are dense
It is reduced to 1g raw medicinal herbs/ml, 0-4 DEG C of refrigerated overnight analyses glue, filters, HPD400 model macroporous adsorbing resin for purification on filtrate, successively
With water, 30% ethyl alcohol, 50% ethyl alcohol and 85% ethanol elution, each elution position is obtained, is dried with spray drying process, collected
Spray dried powder to get.Wherein based on steroid saponin constituents, steroid saponin total content is greater than 90% at 85% elution position,
Up to trilliaceae extract.The medicinal extract at the elution position of above-mentioned trilliaceae extract both 85% ethyl alcohol is added to the 100- of equivalent
200 mesh silica gel are mixed thoroughly, and 50 DEG C of decompressions volatilize solvent, grind well, dry method loading, and (200-300 mesh, silica gel amount are upper normal pressure silica gel column
20 times of medicinal extract), chloroform is successively used after loading: methanol: water gradient elution mainly collects chloroform: methanol: water 65
: 35: 1 elution fraction, with pennogenin -3-O- α-L- rhamnopyranosyl in above-mentioned Trillin substance (1 →
2)-β-D-Glucose glycosides, pennogenin -3-O- α-L- rhamnopyranosyl (1 → 4)-[α-L- rhamnopyranosyl (1 →
2)]-β-D-Glucose glycosides and pennogenin -3-O- α-L- rhamnopyranosyl (1 → 4)-α-L- rhamnopyranosyl (1 →
4)-[α-L- rhamnopyranosyl (1 → 2)] 3 kinds of saponin(es of-β-D-Glucose glycosides are detected as reference substance TLC or HPLC, are merged
Corresponding fraction.Above-mentioned fraction is evaporated in 65 DEG C of reduced pressures, is isolated and purified with preparative high-performance liquid chromatographic technology, is used ethanol water
50-95% elution, eluent are dried through appropriate concentration spray drying, spray dried powder are collected, with 10-95% ethyl alcohol knot
It is brilliant and recrystallize to get.Pennogenin -3-O- α-L- rhamnopyranosyloxyhy in Trillin substance is prepared using the above method
The yield of glycosyl (1 → 2)-β-D-Glucose glycosides is 1.5-2.6g/kg, is 99.3% through HPLC detection purity；Pennogenin-
The yield of 3-O- α-L- rhamnopyranosyl (1 → 4)-[α-L- rhamnopyranosyl (1 → 2)]-β-D-Glucose glycosides is 0.8-
1.9g/kg is 99.1% through HPLC detection purity；Pennogenin -3-O- α-L- rhamnopyranosyl (1 → 4)-α-L- pyrans
Rhamnopyranosyl (1 → 4)-[α-L- rhamnopyranosyl (1 → 2)] yield that the yield of-β-D-Glucose glycosides respectively is is
0.3-0.9g/kg is 98.8% through HPLC detection purity；Chromatographic condition UItimate RXB-C18 chromatographic column (4.6mm ×
150mm, 3 μm), mobile phase is acetonitrile-water (10%-100% gradient elution), Detection wavelength 203nm, flow velocity 1.0ml/
Min, 30 DEG C of column temperature, 20 μ l of sample volume.
Embodiment 4
Inventor causes myocardial ischemia in rats experimental model using coronary ligation, from electrocardiogram, myocardial infarct size, serum flesh
Vigor of acid kinase (CK) and serum lactic dehydrogenase (SLDH) (LDH) etc. research confirms 85% ethanol elution object of trilliaceae to cardiac muscle
Ischemic has significant preventive and therapeutic effect and stronger bioactivity.
1 medicinal material extract: by dry trilliaceae root and rhizome 30kg, being ground into coarse powder, extracts 3 with 70% alcohol reflux
Secondary, 2.0 hours every time, filtration, combined extract was recovered under reduced pressure, and get dry extract (total extract).By AB-8 after dry cream water is suspended
The separation of type macroporous absorbent resin, successively uses water, 30% ethyl alcohol, 60% ethyl alcohol, 85% ethanol elution, obtains each elution position.It will
70% ethanol total extract of trilliaceae (1g is equivalent to crude drug 6.7g), water elution object (1g is equivalent to crude drug 12.2g), 30% ethyl alcohol
Eluate (1g is equivalent to crude drug 83.3g), 60% ethanol elution object (1g is equivalent to crude drug 55.6g), 85% ethanol elution object (1g
It is equivalent to crude drug 26.7g), test liquid is prepared with 2% polysorbate aqueous solution respectively.
2 animal packets and administration: rat 60, by weight be randomly divided into sham-operation group, model group, positive group (the heart difficult to understand
Urapidil is 60mg/kg), 85% ethanol elution object high dose group be 88mg/kg, 85% ethanol elution object middle dose group is
Equal amount of distilled water is given in 44mg/kg, dosage 20ml/kg, sham-operation group, model group filling, 1 time a day, continuous 14 days.
The preparation of 3 models: after last dose 1h, 3% yellow Jackets 45mg/kg anesthetized animal is injected intraperitoneally, by animal
Dorsal position is fixed on mouse platform, cuts off throat's skin, blunt separation subcutaneous fascia, muscle, finds tracheae, row endotracheal intubation,
And connect animal respirator assisted respiartion.In chest unhairing, disinfection, along left mid-clavicular line longitudinal incision skin about 2cm, in the 4th, 5
Intercostal blunt separation muscle layer opens thoracic cavity, digs out heart with from round coring ring.Pulmonary conus and left auricle of heart lower edge can
To see left coronary artery descending anterior branch initial part, at 1~2mm of left auricle of heart lower edge, ligatured with 6/0 suture.Depth of needle control exists
2mm or so, width are about 2mm, and heart is put back to thoracic cavity after ligation, close chest with haemostatic clamp rapidly after excluding the air in thoracic cavity
Chamber.Sham-operation group, which only threads, not to ligature.
4 observation index:
4.1 II lead electrocardiogram Animal Anesthesia dorsal positions are fixed on mouse platform, and four limbs are subcutaneously inserted the needle-shaped electricity of electrocardiogram
Pole, record normal II lead electrocardiogram and coronary ligation half an hour after electrocardiogram (the ECG ST section back of a bow is obviously raised after ligation,
And the myocardium color of ligation shoals to represent and ligature successfully), the variation of electrocardiogram, ST sections of 30min and T wave after observation ligation.
After 4.2 myocardial infarct size abdominal aortic bloods, heart is taken out rapidly, is rinsed well in heart with PBS buffer solution
Remaining blood sucks moisture with filter paper, heart is taken out after freezing 8min in -20 DEG C of refrigerators, rapidly by part under left ventricle ligature
Myocardium to be cut into the sheet that thickness is about 2mm in parallel, sheet cardiac muscle restores to be protected from light dye in the 1%TTC buffer that room temperature postposition is now matched
During which color, and 37 DEG C of water bath with thermostatic control 15min vibrate 4 times sufficiently to dye.Cardiac muscular tissue is so placed in 10% Fu Er after dyeing
It is impregnated in Malin for 24 hours, normal myocardium is contaminated for red, and infarct cardiac muscle is not colored.Cardiac muscular tissue's sequence is taken pictures, is made
The myocardial infarction area area taken pictures is analyzed with Image Pro Plus6.0 medical image analysis system, uses count
And measure abiects tool calculates Infarct area and entire myocardial area, infarction size=Infarct area/heart
The gross area × 100%.
Abdominal aortic blood 6ml after the detection myocardial infarction 3h of 4.3 CK, LDH vigor, 4000r/min is centrifuged after standing 2h
20min takes supernatant.The vigor of kit detection CK, LDH.
5 statistical processing methods:
As a result it usesIt indicates, with different time point measured values and before ligaturing, the difference of basic value indicates electrocardiogram.
Comparison among groups are carried out using SPSS13.0 statistical software one-way analysis of variance method.
6 test results:
The influence that 6.1 trilliaceae, 85% ethanol elution object changes ST sections of rats with myocardial ischemia II lead electrocardiogram and T wave
It the results are shown in Table 1, table 2.Sham-operation group is compared, and model group rats are 5 after coronary ligation, 10,15,30min electrocardiogram
ST sections, T wave significantly increases.Compared with model group, positive drug group, 85% ethanol elution object high dose group of trilliaceae and prolong age
Careless 85% ethanol elution object middle dose group, which can significantly fight ligation coronary artery, leads to raising (p < 0.05 or the p of ST sections of rat, T wave
< 0.01).
Influence of 6.2 trilliaceae, the 85% ethanol elution object to rats with myocardial ischemia myocardial infarct size
It the results are shown in Table 3.Compared with sham-operation group, model group rats myocardial infarct size is significantly increased；With model group phase
Compare, positive drug group, 85% ethanol elution object high dose group of trilliaceae, 85% ethanol elution object middle dose group of trilliaceae can be shown
Writing reduces myocardial infarct size (p < 0.01), illustrates positive drug group, 85% ethanol elution object high dose group of trilliaceae, trilliaceae
85% ethanol elution object middle dose group can increase ischemic region blood supply, reduce myocardial ischemia damage.
3 trilliaceae of table, 85% ethanol elution object to rats with myocardial ischemia myocardial infarct size influence (N=11)
Note: compared with sham-operation group, ##P < 0.01；Compared with model group, * * P < 0.01.
6.3 influence of the 85% ethanol elution object of trilliaceae to rats with myocardial ischemia sero-enzyme CK, LDH
It the results are shown in Table 4.Compared with sham-operation group, CK, LDH vigor are significant in serum after ligaturing coronary artery for model group rats
It increases；Compared with model group, positive drug group, 85% ethanol elution object high dose group of trilliaceae, 85% ethanol elution of trilliaceae
CK, LDH activity significantly reduce (p < 0.01) in the serum of object middle dose group, prompt positive drug group, 85% ethanol elution of trilliaceae
Object high dose group, 85% ethanol elution object middle dose group of trilliaceae have protective effect to the ischemic injuries of cardiac muscle.
4 trilliaceae of table, 85% ethanol elution object to rats with myocardial ischemia sero-enzyme CK, LDH vigor influence (N=
11)
Note: compared with sham-operation group, ##P < 0.01, compared with model group, * * P < 0.01.
7 experiment brief summaries:
Myocardial ischemia is due to caused by the supply and demand disequilibrium of cardiac blood and oxygen, is that the ischemic heart diseases such as coronary heart disease occur
Main pathophysiological mechanism.In the related experimental study of myocardial ischemia, the ECG change of experimental animal is important observation
One of index, the variation of ST sections and T wave height be myocardial ischemia earliest period occur, most sensitive and exact index one, commonly use
In judge myocardial infarction and ischemia model whether duplication successful and the drug effect that resists myocardial ischemia of evaluation drug.Electrocardiogram institute when myocardial ischemia
It is that the cardiac muscle cell depolarized due to external membrane of heart ischemic region inner part and non-ischemic region cardiac muscle are thin that the ST section shown, which is raised,
Caused by the transient current of injury formed between born of the same parents, the raising of T wave is due to subendocardiac muscle ischemic.Therefore, usually will
Main indicator of the changing value of ST sections and T wave as evaluation myocardial ischemia in rats damage.
CK is the important kinases for having direct relation with intracellular energy transhipment, contraction of muscle, ATP regeneration, it is reversible
Catalysis creatine and ATP between the phosphoryl that turns react.It is mainly distributed in skeletal muscle, cardiac muscle and brain cell.Its major function
Are as follows: under atriphos (ATP) effect, it is catalyzed creatine phosphate, forms phosphocreatine and adenosine diphosphate (ADP) (ADP), and
The energy-rich phosphate bond of catalytic phosphatase creatine is transferred to adenosine diphosphate (ADP) (ADP) and generates atriphos (ATP), and forms creatine.
Cell membrane can be released into blood by different degrees of damage, creatine kinase from damaged cell after myocardial ischemia, make flesh in serum
Acid kinase is horizontal to be improved rapidly, and creatine kinase is the important blood biochemistry index of myocardial infarction.
LDH is a kind of oligomerization enzyme, can be catalyzed the reversible transition of lactic acid and pyruvic acid.The enzyme is widely distributed in myocardium, red thin
In born of the same parents, skeletal muscle and liver, its content in cell liquid is very rich, thus the reaction being catalyzed is very fast, is cell
Important biomolecule enzyme in respiratory chain.When histanoxia, cell glycolysis is vigorous, and pyruvic acid is transformed into cream under LDH effect
The process of acid is reinforced, and the raising of LDH activity means that anerobic glycolysis process is reinforced, and aerobic oxidation process is suppressed, energy supply
Obstacle.So in serum LDH activity can reflecting myocardium ischemic injuries range and severity, activity it is higher, reflect the heart
Injury of muscle is heavier.
Originally experimental studies have found that, after the ligation of rat left coronary artery descending anterior branch, ECG ST section, T wave are significantly increased, cardiac muscle stalk
Plug range dramatically increases, and CK, LDH vigor are significantly raised in serum, prompts myocardial ischemia modeling success.85% ethyl alcohol of trilliaceae is washed
De- object high dose group, 85% ethanol elution object middle dose group of trilliaceae can significantly reduce the exception of ST sections of rat model and T wave
It raises, reduces the myocardial infarct size of rats with myocardial ischemia, inhibit the vigor of CK, LDH in serum, prompt 85% ethyl alcohol of trilliaceae
Eluate has certain preventive and therapeutic effect to coronary ligation Myocardial Ischemia.
Embodiment 5
Trilliaceae constituent analysis and quality standard research, seminar find therein in the research to trilliaceae ingredient
Trilliaceae pennogenin B3(pennogenin -3-O- α-L- rhamnopyranosyl (1 → 2)-β-D-Glucose glycosides), B1(inclined promise soap
Aglycon -3-O- α-L- rhamnopyranosyl (1 → 4)-[α-L- rhamnopyranosyl (1 → 2)]-β-D-Glucose glycosides), B2(partially
Promise sapogenin -3-O- α-L- rhamnopyranosyl (1 → 4)-α-L- rhamnopyranosyl (1 → 4)-[α-L- rhamnopyranosyl
(1 → 2)]-β-D-Glucose glycosides) content is larger, and it is specificity ingredient and characteristic chemical constituent in trilliaceae, and at 203nm
There is UV absorption, directly can carry out assay with UV detector, method is easy.Trilliaceae is measured using HPLC method herein
Middle Trillin B2、B1、B3Content, provide foundation to study and formulating the quality standard of trilliaceae medicinal material.
B3: pennogenin -3-O- α-L- rhamnopyranosyl (1 → 2)-β-D-Glucose glycosides (pennogenin-3-O-
α-L-rhamnopyranosyl- (1 → 2)-β-D-glucopyranoside), molecular structural formula is shown in Fig. 1.
B1: pennogenin -3-O- α-L- rhamnopyranosyl (1 → 4)-[α-L- rhamnopyranosyl (1 → 2)]-β -
D-Glucose glycosides (pennogenin-3-O- α-L-rhamnopyranosyl- (1 → 4)-[α-L-rhamnopyranosyl- (1
→ 2)]-β-D-glucopyranoside), molecular structural formula is shown in Fig. 2.
B2: pennogenin -3-O- α-L- rhamnopyranosyl (1 → 4)-α-L- rhamnopyranosyl (1 → 4)-[α-L-
Rhamnopyranosyl (1 → 2)]-β-D-Glucose glycosides (pennogenin-3-O- α-L-rhamnopyranosyl- (1 → 4)-α-
L-rhamnopyranosyl- (1 → 4)-[α-L-rhamnopyranosyl- (1 → 2)]-β-D-glucopyranoside),
Molecular structural formula is shown in Fig. 3.
Reference substance Trillin B2(pennogenin -3-O- α-L- rhamnopyranosyl (1 → 4)-α-L- rhamnopyranosyloxyhy
Glycosyl (1 → 4)-[α-L- rhamnopyranosyl (1 → 2)]-β-D-Glucose glycosides), B1(pennogenin -3-O- α-L- pyrans
Rhamnopyranosyl (1 → 4)-[α-L- rhamnopyranosyl (1 → 2)]-β-D-Glucose glycosides), B3(pennogenin -3-O- α-L-
Rhamnopyranosyl (1 → 2)-β-D-Glucose glycosides) it is made by oneself by this laboratory, it is measured by efficient liquid phase areas of peak normalization method
Mass fraction 99%.Trilliaceae medicinal material picks up from Hubei Province Shennongjia, identifies through Jiangxi University of Traditional Chinese Medicine associate professor Zuo Yueming
For Liliaceae Trillium plant trilliaceae Trillium tschonoskii Maxim..Acetonitrile is that (α Cygni friend is raw for chromatographically pure
Object medical technology Co., Ltd), water is secondary distilled water (self-control).
Sample measurement weighs trilliaceae medicinal powder (crossing No. 4 sieves, low temperature drying to constant weight) about 1.0g, molten according to test sample
Prepared by the preparation method of liquid, accurate 20 μ l of pipette samples solution, injection high performance liquid chromatograph measurement, according to external standard two-point method meter
Calculate the Trillin B in sample2、B1、B3Content be respectively 0.13%, 0.19% and 0.28% (n=3).
The Primary Study of HPLC method measurement trilliaceae constituents absorbed into blood
This seminar has also carried out the Contained Serum chromatographic fingerprinting research of trilliaceae, but only tentatively establishes drug containing blood
Clear preparation method and detection method, primarily determining that trilliaceae enters blood component is B1(pennogenin -3-O- α-L- rhamnopyranosyloxyhy
Glycosyl (1 → 4)-[α-L- rhamnopyranosyl (1 → 2)]-β-D-Glucose glycosides) and B3(pennogenin -3-O- α-L- pyrans
Rhamnopyranosyl (1 → 2)-β-D-Glucose glycosides), to keep the quality evaluating method of trilliaceae more perfect, for its clear drug effect
Material base and the mechanism of action provide wider scientific basis.
Embodiment 6
Inventor causes myocardial ischemia in rats experimental model using coronary ligation, from electrocardiogram, myocardial infarct size, serum flesh
Vigor of acid kinase (CK) and serum lactic dehydrogenase (SLDH) (LDH) etc. research confirms the 85% effective portion of ethanol elution object of trilliaceae
Pennogenin -3-O- α-L- rhamnopyranosyl (1 → 2)-β-D-Glucose glycosides, pennogenin -3-O- α-L- in position
Rhamnopyranosyl (1 → 4)-[α-L- rhamnopyranosyl (1 → 2)]-β-D-Glucose glycosides and pennogenin -3-O- α-L-
Rhamnopyranosyl (1 → 4)-α-L- rhamnopyranosyl (1 → 4)-[α-L- rhamnopyranosyl (1 → 2)]-β-D-Glucose
Glycosides has significant preventive and therapeutic effect and stronger bioactivity to myocardial ischemia.
1 drug and reagent: (pennogenin -3-O- α-L- rhamnopyranosyl (1 → 2) Portugal-β-D- of Trillin 1
Polyglycoside), (pennogenin -3-O- α-L- rhamnopyranosyl (1 → 4)-[α-L- rhamnopyranosyl (1 of Trillin 2
→ 2)]-β-D-Glucose glycosides), (pennogenin -3-O- α-L- rhamnopyranosyl (1 → 4)-α-L- pyrrole of Trillin 3
Mutter rhamnopyranosyl (1 → 4)-[α-L- rhamnopyranosyl (1 → 2)]-β-D-Glucose glycosides) it is white powder, by the author's reality
It is voluntarily isolated to test room, mass fraction is greater than 98%.
2 experimental methods:
2.1 animal packets and administration: rat 90, by weight be randomly divided into sham-operation group, model group, positive group (it is difficult to understand
Sodium Ferulate capsule strength is high for 6mg/ml), 1 high dose group of glycosides (29.14mg/kg), 1 middle dose group of glycosides (14.57mg/kg), glycosides 2
Dosage group (17.14mg/kg), 2 middle dose group of glycosides (8.86mg/kg), 3 high dose group of glycosides (26.86mg/kg), 3 middle dose group of glycosides
(14.28mg/kg), every group of 10 mouse.Dosage is 10ml/kg, and sham-operation group, model group, which fill, gives equal amount of distilled water, daily 1
It is secondary, continuous 14 days.The preparation of 2.2 models: after last dose 1h, being injected intraperitoneally 3% yellow Jackets 45mg/kg anesthetized animal,
Supine position is fixed on mouse platform, throat's skin is cut off, blunt separation subcutaneous fascia, muscle find tracheae, and promoting the circulation of qi pipe is inserted
Guan Shu, and connect animal respirator assisted respiartion.In chest unhairing, disinfection, along left mid-clavicular line longitudinal incision skin about 2cm,
4th, 5 intercostal blunt separation muscle layer open thoracic cavity, dig out heart with from round coring ring.In pulmonary conus and left auricle of heart
Lower edge can see that left coronary artery descending anterior branch initial part, at 1~2mm of left auricle of heart lower edge, be ligatured with 6/0 suture.Depth of needle
In 2mm or so, width is about 2mm for control, and heart is put back to thoracic cavity after ligation, uses haemostatic clamp rapidly after excluding the air in thoracic cavity
Close thoracic cavity.Sham-operation group, which only threads, not to ligature.
2.3 observation index:
2.3.1 II lead electrocardiogram Animal Anesthesia dorsal position is fixed on mouse platform, and four limbs are subcutaneously inserted the needle-shaped electricity of electrocardiogram
Pole, record normal II lead electrocardiogram and coronary ligation half an hour after electrocardiogram (the ECG ST section back of a bow is obviously raised after ligation,
And the myocardium color of ligation shoals to represent and ligature successfully), the variation of electrocardiogram, ST sections of 30min and T wave after observation ligation.
2.3.2 taking out heart rapidly after myocardial infarct size abdominal aortic blood, heart is rinsed well with PBS buffer solution
Interior remaining blood, sucks moisture with filter paper, heart is taken out after freezing 8min in -20 DEG C of refrigerators, rapidly by left ventricle ligature lower part
Flesh of diverting one's attention is cut into the sheet that thickness is about 2mm in parallel, and sheet cardiac muscle restores to be protected from light in the 1%TTC buffer that room temperature postposition is now matched
During which dyeing, and 37 DEG C of water bath with thermostatic control 15min vibrate 4 times sufficiently to dye.Cardiac muscular tissue is so placed in 10% good fortune after dyeing
It is impregnated for 24 hours in your Malin, normal myocardium is contaminated for red, and infarct cardiac muscle is not colored.Cardiac muscular tissue's sequence is taken pictures,
The myocardial infarction area area taken pictures is analyzed using Image Pro Plus6.0 medical image analysis system, uses count
And measure abjects tool calculates Infarct area and entire myocardial area, infarction size=Infarct area/entire
The myocardium gross area × 100%.
2.3.3 abdominal aortic blood 6ml after the detection myocardial infarction 3h of CK, LDH vigor, stand 2h after 4000r/min from
Heart 20min takes supernatant.The vigor of kit detection CK, LDH.
2.4 statistical processing methods
As a result it usesIt indicates, with different time point measured values and before ligaturing, the difference of basic value indicates electrocardiogram.
Comparison among groups are carried out using SPSS13.0 statistical software one-way analysis of variance method.
3 experimental results:
The influence that 3.1 Trillins change ST sections of rats with myocardial ischemia II lead electrocardiogram and T wave
It the results are shown in Table 1, table 2.Compared with sham-operation group, model group rats are 5 after coronary ligation, 10,15,30min electrocardio
ST sections of figure, T wave significantly increase.Compared with model group, positive drug group, Trillin 1 (Trillin 2,3) be high, in
Dosage group, which can significantly fight ligation coronary artery, leads to the raising (p < 0.05 or p < 0.01) of ST sections of rat, T wave.
Influence of 3.2 Trillins to rats with myocardial ischemia myocardial infarct size
It the results are shown in Table 3.Compared with sham-operation group, model group rats myocardial infarct size is significantly increased；With model group phase
Compare, positive drug group, Trillin 1 (Trillin 2,3) are high, middle dose group can significantly reduce myocardial infarct size (p
< 0.01 or p < 0.05), it is scarce to illustrate that positive drug group, Trillin 1 (Trillin 2,3) height, middle dose group can increase
The blood supply of blood area reduces myocardial ischemia damage.
3 Trillin of table to rats with myocardial ischemia myocardial infarct size influence (N=8)
Note: compared with sham-operation group, ##P < 0.01；Compared with model group, * P < 0.05, * * P < 0.01.
Influence of 3.3 Trillins to rats with myocardial ischemia sero-enzyme CK, LDH
It the results are shown in Table 4.Compared with sham-operation group, CK, LDH vigor are significant in serum after ligaturing coronary artery for model group rats
It increases；Compared with model group, positive drug group, Trillin 1 (Trillin 2,3) be high, CK in middle dose group serum,
LDH activity significantly reduces (p < 0.05 or p < 0.01), prompt positive drug group, Trillin 1 (Trillin 2,3) it is high,
Middle dose group has protective effect to the ischemic injuries of cardiac muscle.
4 Trillin of table to rats with myocardial ischemia sero-enzyme CK, LDH vigor influence (N=8)
Note: compared with sham-operation group, ##P < 0.01；Compared with model group, * P < 0.05, * * P < 0.01.
4 experiment brief summaries:
Myocardial ischemia is due to caused by the supply and demand disequilibrium of cardiac blood and oxygen, is that the ischemic heart diseases such as coronary heart disease occur
Main pathophysiological mechanism.In the related experimental study of myocardial ischemia, the ECG change of experimental animal is important observation
One of index, the variation of ST sections and T wave height be myocardial ischemia earliest period occur, most sensitive and exact index one, commonly use
In judge myocardial infarction and ischemia model whether duplication successful and the drug effect that resists myocardial ischemia of evaluation drug.Electrocardiogram institute when myocardial ischemia
It is that the cardiac muscle cell depolarized due to external membrane of heart ischemic region inner part and non-ischemic region cardiac muscle are thin that the ST section shown, which is raised,
Caused by the transient current of injury formed between born of the same parents, the raising of T wave is due to subendocardiac muscle ischemic.Therefore, usually will
Main indicator of the changing value of ST sections and T wave as evaluation myocardial ischemia in rats damage.
CK is the important kinases for having direct relation with intracellular energy transhipment, contraction of muscle, ATP regeneration, it is reversible
Catalysis creatine and ATP between the phosphoryl that turns react.It is mainly distributed in skeletal muscle, cardiac muscle and brain cell.Its major function
Are as follows: under atriphos (ATP) effect, it is catalyzed creatine phosphate, forms phosphocreatine and adenosine diphosphate (ADP) (ADP), and
The energy-rich phosphate bond of catalytic phosphatase creatine is transferred to adenosine diphosphate (ADP) (ADP) and generates atriphos (ATP), and forms creatine.
Cell membrane can be released into blood by different degrees of damage, creatine kinase from damaged cell after myocardial ischemia, make flesh in serum
Acid kinase is horizontal to be improved rapidly, and creatine kinase is the important blood biochemistry index of myocardial infarction.
LDH is a kind of oligomerization enzyme, can be catalyzed the reversible transition of lactic acid and pyruvic acid.The enzyme is widely distributed in myocardium, red thin
In born of the same parents, skeletal muscle and liver, its content in cell liquid is very rich, thus the reaction being catalyzed is very fast, is cell
Important biomolecule enzyme in respiratory chain.When histanoxia, cell glycolysis is vigorous, and pyruvic acid is transformed into cream under LDH effect
The process of acid is reinforced, and the raising of LDH activity means that anerobic glycolysis process is reinforced, and aerobic oxidation process is suppressed, energy supply
Obstacle.So in serum LDH activity can reflecting myocardium ischemic injuries range and severity, activity it is higher, reflect the heart
Injury of muscle is heavier.
Originally experimental studies have found that, after the ligation of rat left coronary artery descending anterior branch, ECG ST section, T wave are significantly increased, cardiac muscle stalk
Plug range dramatically increases, and CK, LDH vigor are significantly raised in serum, prompts myocardial ischemia modeling success.(the inclined promise of Trillin 1
Sapogenin -3-O- α-L- rhamnopyranosyl (1 → 2)-β-D-Glucose glycosides), (pennogenin -3-O- α-of Trillin 2
L- rhamnopyranosyl (1 → 4)-[α-L- rhamnopyranosyl (1 → 2)]-β-D-Glucose glycosides) and (the inclined promise of Trillin 3
Sapogenin -3-O- α-L- rhamnopyranosyl (1 → 4)-α-L- rhamnopyranosyl (1 → 4)-[α-L- rhamnopyranosyl (1
→ 2)]-β-D-Glucose glycosides) height, middle dose group can significantly reduce ST sections of rat model and the exception of T wave is raised, aobvious to subtract
The myocardial infarct size of few rats with myocardial ischemia, inhibits the vigor of CK, LDH in serum, prompts 1 (pennogenin of Trillin
Member -3-O- α-L- rhamnopyranosyl (1 → 2)-β-D-Glucose glycosides), (pennogenin -3-O- α-L- pyrrole of Trillin 2
Mutter rhamnopyranosyl (1 → 4)-[α-L- rhamnopyranosyl (1 → 2)]-β-D-Glucose glycosides) and 3 (pennogenin of Trillin
Member -3-O- α-L- rhamnopyranosyl (1 → 4)-α-L- rhamnopyranosyl (1 → 4)-[α-L- rhamnopyranosyl (1 →
2)]-β-D-Glucose glycosides) height, middle dose group have certain preventive and therapeutic effect to coronary ligation Myocardial Ischemia.
Embodiment 7
Pennogenin -3-O- α-L- rhamnopyranosyl (1 → 2)-β-D-Glucose glycosides in Trillin substance,
Pennogenin -3-O- α-L- rhamnopyranosyl (1 → 4)-[α-L- rhamnopyranosyl (1 → 2)]-β-D-Glucose glycosides and
Pennogenin -3-O- α-L- rhamnopyranosyl (1 → 4)-α-L- rhamnopyranosyl (1 → 4)-[α-L- rhamnopyranose
Base (1 → 2)]-β-D-Glucose glycosides spectral data information and parsing.
1. pennogenin -3-O- α-L- rhamnopyranosyl-(1 → 2)-β-D-Glucose glycosides (pennogenin-3-O-
α-L-rhamnopyranosyl- (1 → 2)-β-D-glucopyranoside), colorless needles (methanol), molecular formula is
C39H62O13。1H-NMR(CDCl3, 400MHz): δ 0.76 (3H, d, J=6.0Hz, CH3- 27), 0.83 (3H, s, CH3- 18),
0.87 (3H, s, CH3- 19), 1.04 (3H, brs, CH3- 21), 1.19 (3H, m, Rha CH3-6″)；5.36 (1H, s, H-6),
3.62 (2H, m, H-26), 4.85 (1H, d, Glc H-l '), 5.92 (1H, s, Rha H-1 ").13C-NMR(CDCl3, 100MHz):
δ 36.8 (C-1), 31.4 (C-2), 80.5 (C-3), 37.1 (C-4), 141.4 (C-5), 121.9 (C-6), 33.6 (C-7), 31.3
(C-8), 54.1 (C-9), 36.1 (C-10), 22.4 (C-11), 31.7 (C-12), 43.8 (C-13), 51.3 (C-14), 30.6
(C-15), 90.5 (C-16), 90.3 (C-17), 16.8 (C-18), 19.1 (C-19), 46.9 (C-20), 18.6 (C-21),
110.0 (C-22), 33.9 (C-23), 26.9 (C-24), 30.3 (C-25), 66.9 (C-26), 16.7 (C-27), 99.8 (C-
1 '), 79.2 (C-2 '), 77.9 (C-3 '), 73.7 (C-4 '), 77.1 (C-5 '), 62.7 (C-6 '), 100.8 (C-1 "), 72.3
(C-2 "), 71.8 (C-3 "), 71.7 (C-4 "), 68.5 (C-5 "), 19.2 (C-6 ").Molecular structural formula is shown in Fig. 1.
Pennogenin -3-O- α-L- rhamnopyranosyl (1 → 4) -2. [α-L- rhamnopyranosyl (1 → 2)]-β-D-
Glucoside (pennogenin-3-O- α-L-rhamnopyranosyl- (1 → 4)-[α-L-rhamnopyranosyl- (1 →
2)]-β-D-glucopyranoside), white needles (methanol), molecular formula C45H71O17。1H-NMR(CDCl3, 400MHz): δ
0.76 (3H, d, J=6.0Hz, CH3- 27), 0.83 (3H, s, CH3- 18), 0.87 (3H, s, CH3- 19), 1.04 (3H, brs,
CH3- 21), 1.20 (3H, d, J=6.0Hz, Rha ' CH3- 6), 1.23 (3H, d, J=6.8Hz Rha " CH3- 6), 5.36 (1H,
Brs, H-6), 4.88 (1H, d, Glc H-1), 5.49 (1H, s, Rha ' H-1), 6.17 (1H, s, Rha " H-1).13C-NMR
(CDCl3, 100MHz): δ 36.8 (C-1), 31.3 (C-2), 80.5 (C-3), 37.2 (C-4), 141.4 (C-5), 121.9 (C-
6), 33.6 (C-7), 31.4 (C-8), 54.1 (C-9), 36.1 (C-10), 22.4 (C-11), 31.7 (C-12), 43.8 (C-13),
51.3 (C-14), 30.8 (C-15), 90.5 (C-16), 90.3 (C-17), 16.8 (C-18), 19.1 (C-19), 46.9 (C-20),
18.6 (C-21), 110.2 (C-22), 33.9 (C-23), 26.9 (C-24), 30.3 (C-25), 66.9 (C-26), 16.8 (C-
27), 100.2 (Glc C-1 '), 79.2 (C-2 '), 77.9 (C-3 '), 76.7 (C-4 '), 77.1 (C-5 '), 62.7 (C-6 '),
101.8 (Rha ' C-1 "), 72.3 (C-2 "), 71.8 (C-3 "), 71.0 (C-4 "), 69.7 (C-5 "), 19.2 (C-6 "), 102.5
(Rha " C-1 " '), 72.7 (C-2 " '), 72.4 (C-3 " '), 77.1 (C-4 " '), 70.6 (C-5 " '), 19.2 (C-6 " ').Molecule
Structural formula is shown in Fig. 2.
Claims (11)
1. trilliaceae steroid saponin compound is preparing the application in anti-ischemic heart medicine, which is characterized in that this prolongs
Age grass steroid saponin compound include pennogenin -3-O- α-L- rhamnopyranosyl (1 → 2)-β-D-Glucose glycosides, partially
Promise sapogenin -3-O- α-L- rhamnopyranosyl (1 → 4)-[α-L- rhamnopyranosyl (1 → 2)]-β-D-Glucose glycosides and partially
Promise sapogenin -3-O- α-L- rhamnopyranosyl (1 → 4)-α-L- rhamnopyranosyl (1 → 4)-[α-L- rhamnopyranosyl
At least one of (1 → 2)]-β-D-Glucose glycosides.
2. trilliaceae steroid saponin compound according to claim 1 is preparing answering in anti-ischemic heart medicine
With, which is characterized in that related ischemic heart disease is the coronary heart diseases and angina pectoris and cardiac muscle for being caused and being occurred by myocardial ischemia
Any one of the ischemic heart disease of infarction.
3. trilliaceae steroid saponin compound according to claim 1 is preparing answering in anti-ischemic heart medicine
With, which is characterized in that the preparation formulation of the drug is oral solution, capsule, tablet, injection.
4. trilliaceae steroid saponin compound according to claim 1 is preparing answering in anti-ischemic heart medicine
With, which is characterized in that the drug includes single preparations of ephedrine and compound preparation.
5. trilliaceae steroid saponin compound according to claim 1 is preparing answering in anti-ischemic heart medicine
With, which is characterized in that the crude drug source of the trilliaceae steroid saponin compound is Liliaceae Trillium plant trilliaceae
Trillium tschonoskii Maxim.。
6. a kind of trilliaceae extract is preparing the application in anti-ischemic heart medicine, which is characterized in that the trilliaceae
Extract the preparation method comprises the following steps: by trilliaceae pulverizing medicinal materials at coarse powder, impregnate 1-2 hours with 50-70% ethyl alcohol, 8-12 times measure it is molten
Agent heating and refluxing extraction 3 times, 2 hours every time, filtering, merging filtrate；Above-mentioned filtrate is concentrated under reduced pressure into 1g crude drug in 65-70 DEG C
Material/ml, 0-4 DEG C of refrigerated overnight analyse glue, filtering, and filtrate recycles to obtain the upper macroporous absorbent resin of medicinal extract dilution with extracting n-butyl alcohol again
Enrichment or the directly upper macroporous adsorbing resin for purification of filtrate, successively use water, 30% ethyl alcohol, 60% ethyl alcohol and 85% ethanol elution, obtain each
Position is eluted, is dried with freeze-drying or spray drying, freeze-dried powder or spray dried powder are collected, wherein 85% ethyl alcohol is washed
De- position product is the trilliaceae extract.
7. trilliaceae extract according to claim 6 is preparing the application in anti-ischemic heart medicine, feature
It is, the model AB-8 type or D-101 type or HPD-400 of the macroporous absorbent resin.
8. trilliaceae extract according to claim 6 is preparing the application in anti-ischemic heart medicine, feature
It is, related ischemic heart disease is that the coronary heart diseases and angina pectoris for being caused and being occurred by myocardial ischemia and myocardial infarction are formed
Ischemic heart disease any one.
9. trilliaceae extract according to claim 6 is preparing the application in anti-ischemic heart medicine, feature
It is, the preparation formulation of the drug is oral solution, capsule, tablet, injection.
10. trilliaceae extract according to claim 6 is preparing the application in anti-ischemic heart medicine, feature
It is, the drug includes single preparations of ephedrine and compound preparation.
11. trilliaceae extract according to claim 6 is preparing the application in anti-ischemic heart medicine, feature
It is, the crude drug source of the trilliaceae extract is Liliaceae Trillium plant trilliaceae Trillium tschonoskii
Maxim.。
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| CN106138627A (en) * | 2016-07-30 | 2016-11-23 | 张忠立 | The preparation method of monomeric substance and application in preparation treatment Alzheimer disease drugs thereof in China's Trillium or Paris Linnaeus(Paris L.) medical material |
| CN106421286A (en) * | 2016-10-25 | 2017-02-22 | 西安交通大学 | Process for separating and purifying trillium steroid saponin by using macroporous resin |
| CN107693663A (en) * | 2017-10-20 | 2018-02-16 | 张忠立 | Application of the trilliaceae steroid saponin effective constituents in anti-cerebral ischemia reperfusion injury and its apoplexy sequela medicine is prepared |
| CN109438548B (en) * | 2018-12-01 | 2021-07-06 | 中国科学院昆明植物研究所 | Preparation method of paris polyphylla pennogenin Pb |
| CN114588225B (en) * | 2021-12-20 | 2023-09-15 | 嘉文丽（福建）化妆品有限公司 | Extraction and conversion method of saponin in longhairy antenoron herb |
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| CN105168235A (en) * | 2015-10-22 | 2015-12-23 | 江西中医药大学 | Preparation methods of trillium saponin substance and preparation thereof as well as application of trillium saponin substance and preparation thereof in preparation of medicine for treating senile dementia and Alzheimer's disease |
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| CN107648430A (en) * | 2015-10-22 | 2018-02-02 | 江西中医药大学 | A kind of preparation method of the effective constituents of steroid saponin containing trilliaceae capsule preparations and its application in various kinds of drug is prepared |
Non-Patent Citations (5)
| Title |
|---|
| Steroids from the Underground Parts of Trillium kamtschaticum;Masateru Ona et al;《Chem. Pharm. Bull.》;20031231;第51卷(第11期);第1328-1331页 |
| 头顶一颗珠对大鼠缺血再灌注后左心室功能及抗氧化酶的影响;袁杰;《临床合理用药》;20090630;第2卷(第11期);第31-32页，尤其是[摘要]方法、结论 |
| 延龄草属植物的化学成分及药理活性研究进展;喻玲玲等;《中成药》;20080930;第30卷(第9期);第1350-1354页 |
| 延龄草根及根茎化学成分研究;贾兰婷等;《中国实验方剂学杂志》;20140331;第20卷(第6期);第99-102页，尤其是[2 提取与分离]及[3 结构鉴定] |
| 延龄草根及根茎的化学成分研究(III);张忠立等;《中药材》;20131130;第36卷(第11期);第1779-1782页 |
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