CN107141305B - It is a kind of to be isolated from the blue novel Crow alkane type diterpene-kind compound for spending radix scutellariae and its in the purposes prepared in inverase - Google Patents
It is a kind of to be isolated from the blue novel Crow alkane type diterpene-kind compound for spending radix scutellariae and its in the purposes prepared in inverase Download PDFInfo
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- C07D493/00—Heterocyclic compounds containing oxygen atoms as the only ring hetero atoms in the condensed system
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Abstract
The invention belongs to technical field of phytochemistry, and in particular to a kind of to be isolated from the blue novel Crow alkane type diterpene-kind compound for spending radix scutellariae and its in the purposes prepared in inverase.The neo clerodane diterpenoids class compound has structure shown in Formulas I:Wherein R1Selected from Ac, Bz, R2Selected from OH, OAc or OBz, R3Selected from H, OAc or OBz.Disease caused by formula Compound I can be used for treating and/or preventing being infected by HIV-1.
Description
Technical field
The invention belongs to technical field of phytochemistry, and in particular to a kind of novel Crow alkane type two for being isolated from blue flower radix scutellariae
Terpenoid and its preparing the purposes in inverase.
Background technique
Labiatae (Lamiaceae) Scutellaria (Scutellaria) plant is with being distributed widely in Perenniporia martius mountain range
Area, throughout Europe, North America and East Asia, there are about more than 360 a species.There are 102 kinds what China was expressly recited, 50 mutation, mostly
It is medicinal.Wherein 2010 version " Chinese Pharmacopoeia " include 2 kinds: radix scutellariae (Scutellaria baicalensis) and Sculellaria barbata
(Scutellaria barbata), radix scutellariae cure mainly damp-warm syndrome, summer-heat and damp, chest tightness and emesis, damp and hot feeling of fullness, dysentery, jaundice, cough with lung heat,
High fever is tired of cough, blood-head tells nosebleed, carbuncle sore tumefacting virus and threatened abortion.Modern pharmacological studies have shown that radix scutellariae platymiscium is with anti-oxidant, anti-
Tumour, liver protection, anti-inflammatory, anticonvulsion, antibacterial, it is antiviral the effects of.Terpene and phenolic compound are mainization of radix scutellariae platymiscium
It studies point, wherein Diterpenes have inhibits the duplication of EBV virolysis well, and it is antitumor, resist food and to lipopolysaccharide-induced
Nitric oxide has generated inhibiting effect isoreactivity.
Indigo plant flower radix scutellariae (Scutellaria formosana N.E.Brown) is that kind Labiatae (Lamiacea) Scutellaria is more
Year raw herbaceous plant, rhizome is closely wooden, tool fibrous fibrous root.The plant originates from Guangdong, Jiangxi, Fujian, the ground such as Yunnan.It closes at present
Volatile oil component identification and its crude extract activity research, knot are only limitted in the chemical component of blue flower radix scutellariae and the research of pharmacological activity
Fruit shows that blue flower radix scutellariae crude extract has certain antitumor and Anti-HIV-1 Active.According to this platymiscium relevant report, these activity
It may be related with Diterpenoids from bulbus.The neo clerodane diterpenoids class compound isolated from blue flower radix scutellariae has not been reported, also not
See that such compound is reported in anti-HIV-1.
Summary of the invention
The present invention provide a kind of neo clerodane diterpenoids class compound, its stereoisomer, tautomer, prodrug or its
Pharmaceutically acceptable salt, it is characterised in that the neo clerodane diterpenoids class compound has structure shown in Formulas I:
Wherein R1Selected from Ac, Bz, R2Selected from OH, OAc or OBz, R3Selected from H, OAc or OBz.
The present invention provides another preferred embodiment, the preferably following compound II-VIII of compound of formula I:
Another embodiment of the present invention provides the method for prepare compound II-VIII simultaneously a kind of, it is characterised in that packet
Include following steps:
(1) the indigo plant flower radix scutellariae herb to dry in the shade is crushed, heats 50-60 DEG C with ethanol solution and extract 2-4 times, extracts 1- every time
Crude extract is concentrated under reduced pressure to obtain in 3h, combined extract；
(2) after suspension being made with the dilution of suitable water in the crude extract that step (1) obtains, successively with isometric petroleum
Ether extracts 3-5 times, merges petroleum ether phase, medicinal extract is concentrated under reduced pressure to obtain；
(3) medicinal extract for obtaining step (2), first through depressurizing silica gel column chromatography, using petroleum ether-ethyl acetate mixed solvent
Gradient elution is carried out as eluant, eluent, gradient is respectively 100:0,90:10,80:20,70:30,60:40,50:50,40:
60,30:70,20:80,10:90,0:100, each gradient collect two column volumes, are divided into 10 components according to polarity size,
It is component 1 that middle gradient 100:0~90:10, which is afforded, and it is component 2 that gradient 90:10~80:20, which is afforded, gradient 80:
It is component 3 that 20~70:30, which is afforded, and it is component 4, gradient 60:40~50:50 that gradient 70:30~60:40, which is afforded,
Affording is component 5, and it is component 6 that gradient 50:50~40:60, which is afforded, and gradient 40:60~30:70 is afforded
It is component 7, it is component 8 that gradient 30:70~20:80, which is afforded, and it is component that gradient 20:80~10:90, which is afforded,
It is component 10 that 9, gradient 10:90~0:100, which are afforded, and wherein for component 4 first through normal-phase silica gel column chromatography, eluant, eluent is petroleum
Ether: ethyl acetate=15:1-5:1 mixed solvent elutes 2-5 column volume, again through Sephadex LH-20 after reduced pressure
Gel filtration chromatography, eluant, eluent CHCl3: the mixed solvent of MeOH=1:1 elutes 3-6 column volume, passes through again after reduced pressure
High-efficient liquid phase chromatogram HPLC preparation, chromatographic column are Waters C18,9.4 × 250mm, 7 μm, flow velocity 2mL/min, mobile phase
For MeOH:H2O=55:45, successively obtains compound II, IV, VI, and component 5 first through Sephadex LH-20 gel filtration chromatography, is washed
De- agent is MeOH, elutes 3-6 column volume, prepares again through high-efficient liquid phase chromatogram HPLC after reduced pressure, chromatographic column Waters
C18,9.4 × 250mm, 7 μm, flow velocity 2mL/min, mobile phase MeOH:H2O=75:25, successively obtain compound III,
VIII, component 6 are prepared through high-efficient liquid phase chromatogram HPLC, and chromatographic column is Waters C18,9.4 × 250mm, 7 μm, and flow velocity is
2mL/min, mobile phase MeCN:H2O=75:25 successively obtains compound VII, V.
Above-mentioned preparation method, ethanol solution described in step (1) are the ethanol solution that volume fraction is 75-95%, are used
Amount uses 2-3L ethanol solution for every kilogram of blue flower radix scutellariae herb (dry weight).The dosage of water, those skilled in the art in step (2)
The dosage of water can be reasonably selected according to the amount of crude extract, preferably every 100 grams of crude extracts add water 200-300mL, extract every time
The volumetric usage of petroleum ether and the volume of water are identical.
Related to eluant, eluent or mobile phase ratio are volume ratio in chromatographic isolation of the present invention.
Another embodiment of the present invention provides a kind of inverase, it is characterised in that the drug with compound of formula I, its
Stereoisomer, tautomer, prodrug or its pharmaceutically acceptable salt are as effective component.The drug may also include other
Antiviral drugs.The drug may also include pharmaceutic adjuvant, such as pharmaceutically acceptable carrier, excipient or diluent.
Another embodiment of the present invention provides a kind of inverase, it is characterised in that the drug is with compound II-
VIII, its stereoisomer, tautomer, prodrug or its pharmaceutically acceptable salt are as effective component.The drug may be used also
Including other antiviral drugs.The drug may also include pharmaceutic adjuvant, such as pharmaceutically acceptable carrier, excipient or dilution
Agent.
Another embodiment of the present invention provides compound of formula I, its stereoisomer, tautomer, prodrug or its medicine
Acceptable salt is preparing the purposes in inverase on.Especially preparing the purposes in anti-HIV-1 medicines.
Another embodiment of the present invention provide compound II-VIII, its stereoisomer, tautomer, prodrug or
Its pharmaceutically acceptable salt is preparing the purposes in inverase.Especially preparing the purposes in anti-HIV-1 medicines.
Term " pharmaceutically acceptable salt " refers to the addition of atoxic inorganic or organic acid and/or alkali in the present invention
Salt, reference can be made to " Salt selection for basic drugs ", Int.J.Pharm. (1986), 33,201-217.
Specific embodiment
For the ease of a further understanding of the present invention, examples provided below has done more detailed description to it.But
It is that these embodiments are only not supposed to be a limitation to the present invention or implementation principle for better understanding invention, reality of the invention
The mode of applying is not limited to the following contents.
Embodiment 1: the preparation of Crow alkane type diterpene-kind compound II-VIII
(1) indigo plant flower radix scutellariae herb (2.0kg) to dry in the shade is crushed, heats 50-60 DEG C with 75% ethanol solution (6L) and mentions
It takes 4 times, extracts 1h every time, crude extract (about 300g) is concentrated under reduced pressure to obtain in combined extract；
(2) crude extract that step (1) obtains is diluted with suitable water (600mL) after suspension is made, successively with etc. bodies
Long-pending petroleum ether (600mL) extracts 5 times, merges petroleum ether phase, medicinal extract (about 21g) is concentrated under reduced pressure to obtain；
(3) medicinal extract for obtaining step (2), first through depressurizing silica gel column chromatography, using petroleum ether-ethyl acetate mixed solvent
Gradient elution is carried out as eluant, eluent, gradient is respectively 100:0,90:10,80:20,70:30,60:40,50:50,40:
60,30:70,20:80,10:90,0:100, each gradient collect two column volumes, are divided into 10 components according to polarity size,
It is component 1 that middle gradient 100:0~90:10, which is afforded, and it is component 2 that gradient 90:10~80:20, which is afforded, gradient 80:
It is component 3 that 20~70:30, which is afforded, and it is component 4, gradient 60:40~50:50 that gradient 70:30~60:40, which is afforded,
Affording is component 5, and it is component 6 that gradient 50:50~40:60, which is afforded, and gradient 40:60~30:70 is afforded
It is component 7, it is component 8 that gradient 30:70~20:80, which is afforded, and it is component that gradient 20:80~10:90, which is afforded,
It is component 10 that 9, gradient 10:90~0:100, which are afforded, and wherein for component 4 first through normal-phase silica gel column chromatography, eluant, eluent is petroleum
Ether: ethyl acetate=15:1-5:1 mixed solvent elutes 2-5 column volume, again through Sephadex LH-20 after reduced pressure
Gel filtration chromatography, eluant, eluent CHCl3: the mixed solvent of MeOH=1:1 elutes 3-6 column volume, passes through again after reduced pressure
High-efficient liquid phase chromatogram HPLC preparation, chromatographic column are Waters C18,9.4 × 250mm, 7 μm, flow velocity 2mL/min, mobile phase
For MeOH:H2O=55:45, successively obtains compound II (20.0mg), IV (2.8 mg), VI (10.0mg), and component 5 first passes through
Sephadex LH-20 gel filtration chromatography, eluant, eluent MeOH elute 3-6 column volume, again through efficient liquid phase after reduced pressure
Chromatography HPLC preparation, chromatographic column be Waters C18,9.4 × 250mm, 7 μm, flow velocity 2mL/min, mobile phase MeOH:
H2O=75:25 successively obtains compound III (18.9mg), VIII (2.5mg), and component 6 is through high-efficient liquid phase chromatogram HPLC system
Standby, chromatographic column is Waters C18,9.4 × 250mm, 7 μm, flow velocity 2mL/min, mobile phase MeCN:H2O=75:25, according to
It is secondary to obtain compound VII (3.0mg), V (2.7mg).
Compound II-VIII structure is as follows:
Structural Identification data are as follows:
Compound II:(1R, 4R, 5S, 6,8S, 9R, 10R, 12S, 13R) -1,6- diacetoxy -12- benzoyloxy
-4,18；New Crow alkane -15, the 16- lactone of 8,13- diepoxies-is colorless crystalline object, is soluble in chloroform.Through HRESI (+) MS (m/z
593.2366[M+Na]+, theoretical value 593.2357) and determine that its molecular formula is C31H38O10；According to1H,13C and two dimensional NMR
Data determine that its structure, framework types are Crow alkane type, are named as Scuteformoid A,1H and13C NMR data is returned
Category be shown in Table 1. [400MHz (1H), 100MHz (13C), solvent: CDCl3]。
Compound III:(1R, 4R, 5S, 6S, 7R, 8S, 9R, 10R, 12S, 13R) -1,6- diacetoxy -7,12- hexichol
Formyloxy -4,18；New Crow alkane -15, the 16- lactone of 8,13- diepoxies-is colorless crystalline object, is soluble in chloroform.Through HRESI
(+)MS(m/z 713.3372[M+Na]+, theoretical value 713.3374) and determine that its molecular formula is C38H42O12；According to1H,13C and two
Dimension nuclear magnetic resonance data determines that its structure, framework types are Crow alkane type, is named as Scuteformoid E,1H and13C
NMR data ownership be shown in Table 2. [400MHz (1H), 100MHz (13C), solvent: CDCl3]。
Compound IV:(1R, 4R, 5S, 6S, 7R, 8S, 9R, 10R, 12S, 13R) -1,6,7- triacetoxyl group -12- benzene first
Acyloxy -4,18；New Crow alkane -15, the 16- lactone of 8,13- diepoxies-is amorphous powder, is soluble in chloroform.Through HRESI
(+)MS(m/z 651.2428[M+Na]+, theoretical value 651.2412) and determine that its molecular formula is C33H40O12；According to1H,13C and two
Dimension nuclear magnetic resonance data determines that its structure, framework types are Crow alkane type, is named as Scuteformoid F,1H and13C
NMR data ownership be shown in Table 2. [400MHz (1H), 100MHz (13C), solvent: CDCl3]。
Compound V:(1R, 4R, 5S, 6S, 7R, 8S, 9R, 10R, 12S, 13R) -1- hydroxyl -6- acetoxyl group -7,12- two
Benzoyloxy -4,18；New Crow alkane -15, the 16- lactone of 8,13- diepoxies-is amorphous powder, is soluble in chloroform.Through
HRESI(+)MS (m/z 671.2578[M+Na]+, theoretical value 671.2577) and determine that its molecular formula is C36H40O11；According to1H,13C and two dimensional NMR data determine that its structure, framework types are Crow alkane type, are named as Scuteformoid M,
Its1H and13C NMR data ownership be shown in Table 3. [400MHz (1H), 100MHz (13C), solvent: CDCl3]。
Compound VI:(1R, 4R, 5S, 6S, 8S, 9R, 10R, 12S, 13R) -6,12- diacetoxy -1- benzoyloxy
-4,18；New Crow alkane -15, the 16- lactone of 8,13- diepoxies-is colorless crystalline object, is soluble in chloroform.Through HRESI (+) MS (m/z
593.2358[M+Na]+, theoretical value 593.2357) and determine that its molecular formula is C31H38O10；According to1H,13C and two dimensional NMR
Data determine that its structure, framework types are Crow alkane type, are named as Scuteformoid C,1H and13C NMR data is returned
Category be shown in Table 2. [400MHz (1H), 100MHz (13C), solvent: CDCl3]。
Compound VII:(1R, 4R, 5S, 6S, 7R, 8S, 9R, 10R, 12S, 13R) -6,7,12- triacetoxyl group -1- benzene first
Acyloxy -4,18；New Crow alkane -15, the 16- lactone of 8,13- diepoxies-is colorless crystalline object, is soluble in chloroform.Through HRESI
(+)MS(m/z 651.2428[M+Na]+, theoretical value 651.2412) and determine that its molecular formula is C33H40O12；According to1H,13C and two
Dimension nuclear magnetic resonance data determines that its structure, framework types are Crow alkane type, is named as Scuteformoid L,1H and13C
NMR data ownership be shown in Table 3. [400MHz (1H), 100MHz (13C), solvent: CDCl3]。
Compound VIII:(1R, 4R, 5S, 6S, 7R, 8S, 9R, 10R, 12S, 13R) -1,6,12- triacetoxyl group -7- benzene
Formyloxy -4,18；New Crow alkane -15, the 16- lactone of 8,13- diepoxies-is amorphous powder, is soluble in chloroform.Through HRESI
(+)MS(m/z 651.2428[M+Na]+, theoretical value 651.2412) and determine that its molecular formula is C33H40O12；According to1H,13C and two
Dimension nuclear magnetic resonance data determines that its structure, framework types are Crow alkane type, is named as Scuteformoid H,1H and13C
NMR data ownership be shown in Table 3. [400MHz (1H), 100MHz (13C), solvent: CDCl3]。
1. compound II's of table1H-NMR and13C-NMR data [δ (ppm), J (Hz)]
Table 2. compound III, IV, VI's1H-NMR and13C-NMR data [δ (ppm), J (Hz)]
Table 3. compound V, VII, VIII's1H-NMR and13C-NMR data [δ (ppm), J (Hz)]
Embodiment 2
(1) indigo plant flower radix scutellariae herb (2.0kg) to dry in the shade is crushed, heats 50-60 DEG C with 95% ethanol solution (4L) and mentions
It takes 2 times, extracts 3h every time, crude extract (about 310g) is concentrated under reduced pressure to obtain in combined extract；
(2) crude extract that step (1) obtains is diluted with suitable water (930mL) after suspension is made, successively with etc. bodies
Long-pending petroleum ether (930mL) extracts 3 times, merges petroleum ether phase, medicinal extract (about 22g) is concentrated under reduced pressure to obtain；
(3) medicinal extract for obtaining step (2), first through depressurizing silica gel column chromatography, using petroleum ether-ethyl acetate mixed solvent
Gradient elution is carried out as eluant, eluent, gradient is respectively 100:0,90:10,80:20,70:30,60:40,50:50,40:
60,30:70,20:80,10:90,0:100, each gradient collect two column volumes, are divided into 10 components according to polarity size,
It is component 1 that middle gradient 100:0~90:10, which is afforded, and it is component 2 that gradient 90:10~80:20, which is afforded, gradient 80:
It is component 3 that 20~70:30, which is afforded, and it is component 4, gradient 60:40~50:50 that gradient 70:30~60:40, which is afforded,
Affording is component 5, and it is component 6 that gradient 50:50~40:60, which is afforded, and gradient 40:60~30:70 is afforded
It is component 7, it is component 8 that gradient 30:70~20:80, which is afforded, and it is component that gradient 20:80~10:90, which is afforded,
It is component 10 that 9, gradient 10:90~0:100, which are afforded, and wherein for component 4 first through normal-phase silica gel column chromatography, eluant, eluent is petroleum
Ether: ethyl acetate=15:1-5:1 mixed solvent elutes 2-5 column volume, again through Sephadex LH-20 after reduced pressure
Gel filtration chromatography, eluant, eluent CHCl3: the mixed solvent of MeOH=1:1 elutes 3-6 column volume, passes through again after reduced pressure
High-efficient liquid phase chromatogram HPLC preparation, chromatographic column are Waters C18,9.4 × 250mm, 7 μm, flow velocity 2mL/min, mobile phase
For MeOH:H2O=55:45, successively obtains compound II (20.2mg), IV (2.6 mg), VI (11.0mg), and component 5 first passes through
Sephadex LH-20 gel filtration chromatography, eluant, eluent MeOH elute 3-6 column volume, again through efficient liquid phase after reduced pressure
Chromatography HPLC preparation, chromatographic column be Waters C18,9.4 × 250mm, 7 μm, flow velocity 2mL/min, mobile phase MeOH:
H2O=75:25 successively obtains compound III (19.0mg), VIII (2.5mg), and component 6 is prepared through high-efficient liquid phase chromatogram HPLC,
Chromatographic column is Waters C18,9.4 × 250mm, 7 μm, flow velocity 2mL/min, mobile phase MeCN:H2O=75:25, successively
Obtain compound VII (3.2mg), V (2.9mg).
The structural identification data and embodiment 1 of compound II-VIII is consistent.
Embodiment 3: pharmacological activity experiment
Experimental material
Cell: T lymphocytes in human body C8166, MT-4, experimental virus strain HIV-1IIIB。
Cell culture fluid: (FBS) containing 10% fetal calf serum, RPMI-1640 complete medium, 2mM L-Glutamine,
10mM HEPES, 100,000IU/L penicillin, 50 μM of 2 mercapto ethanols, 100 μ g/ml streptomysins.
Reagent: HEPES (N-2 (2-Hydroxyothyl) piperazine-N '-(2-ethanesufonic acid),
MTT [3- (4,5-dimethyl-2-thiazolyl) -2,5-diphenyl-2-H-tetrazolium bromide], paddy ammonia
Amide (Glutamine), penicillin (Penicillin), streptomycin sulphate (Streptomycinsulfate) are purchased from Sigma
Company；RPMI-1640 and fetal calf serum (FBS) are Invitrogen Products；2 mercapto ethanol (2-ME, 2-
It Mercaptoethanol) is Bio-Rad Products；LDH kit (Takara), Poly (A) × [dT]15(Roche),
3H-dTTP (PerkinElmer), MicroscintTM PS(PerkinElmer)。
Experimental method
HIV-1 infectious titration
By HIV-1IIIBStorage liquid makees 4 times of dilutions, point 10 gradients on 96 orifice plates, and 6 repeating holes of every gradient are set simultaneously
Set 6 hole of control wells.50 μ l of C8166 cell is added in every hole, and every hole final volume is 200 μ l.At 37 DEG C, 5%CO2Culture.It mends within 3rd day
Add fresh 100 μ l of RPMI-1640 complete medium, HIV-1 in every hole is observed under inverted microscope within the 7th dayIIIBWhat is induced is thin
Whether born of the same parents' pathological effect (Cytopathic Effect, CPE) has the formation of plasomidum (Syncytium) to determine with every hole；It presses
Reed&Muench method calculates the TCID50 (50%Tissue Culture Infection Dose) of virus.
Cytotoxicity experiment
By 4 × 105A/100 μ L of ml C8166 suspension is mixed from different drug solutions to be measured, and 3 repeating holes are arranged.Together
When setting not drug containing control wells, using AZT as positive drug control.At 37 DEG C, 5%CO2Under conditions of cultivate 3 days, use
MTT colorimetric determination cytotoxicity.ELx800 microplate reader measures OD value, and measurement wavelength is 570nm.CC is calculated50Value (50%
Cytotoxic Concentration), i.e., to 50% normal T-lymphocytes system C8166 generate toxicity when drug concentration.
The Inhibition test of HIV-1 cytopathogenic effect (CPE)
By 8 × 105A/ml normal human's T lymphocyte system C8166 pipettes 50 holes μ L/ and is inoculated into containing 100 holes μ L/ ladder
It spends in 96 porocyte culture plates of doubling dilution drug, the HIV-1 of 50 μ L is then added dropwiseIIIBDilute supernatant, 1300 TCID
50/ hole, every hole final volume are 200 μ l, and 3 repeating holes are arranged.The normal cell controls hole of not drug containing is set simultaneously, with AZT
For positive drug control.At 37 DEG C, 5%CO2Under conditions of cultivate 3 days, (100 ×) count plasomidum under inverted microscope
It is formed.EC50(50%Effective concentration) is compound concentration when inhibiting Syncytium formation 50%.
Statistical method
According to experimental result, line chart is drawn using Origin7.5, sample is calculated by Reed&Muench method and inhibits disease
50% effective concentration (EC of poison50), 50% inhibits cell growth concentration (CC50) and Anti-HIV-1 Active therapeutic index TI value
(Therapeutic index) are as follows: TI=CC50/EC50.
(1) cell grows survival rate (%)=experimental port OD value/value × 100 control wells OD
(2) inhibiting rate (%) of HIV-1 cytopathogenic effect=(1- experimental port plasomidum number/control wells plasomidum number) × 10
Experimental result
4. see the table below to the inhibiting effect result of HIV-1 cytopathogenic effect.The result shows that blue flower radix scutellariae petroleum ether part
Alcohol extracts show preferable Anti-HIV-1 Active, and the neo clerodane diterpenoids separated from petroleum ether part are to HIV-1 disease
Poison has faint inhibiting effect.
The Anti-HIV-1 Active for the crude extract that 4. the compounds of this invention II-VIII of table and step (1) obtain
bTI=CC50/EC50.
cPositive control medicine: 3'- nitrine -3'- deoxythymidine.
Claims (10)
1. a kind of neo clerodane diterpenoids class compound, its stereoisomer, tautomer or its pharmaceutically acceptable salt,
It is characterized in that the neo clerodane diterpenoids class compound has structure shown in Formulas I:
Wherein R1Selected from Ac, Bz, R2Selected from OH, OAc or OBz, R3Selected from H, OAc or OBz.
2. compound of formula I described in claim 1 is selected from following compound II-VIII:
3. a kind of method for preparing neo clerodane diterpenoids class compound II-VIII as claimed in claim 2 simultaneously, feature exist
In including the following steps:
(1) the indigo plant flower radix scutellariae herb to dry in the shade is crushed, heats 50-60 DEG C with ethanol solution and extract 2-4 times, extracts 1-3h every time,
Crude extract is concentrated under reduced pressure to obtain in combined extract；
(2) after suspension being made with the dilution of suitable water in the crude extract that step (1) obtains, successively extracted with isometric petroleum ether
It takes 3-5 times, merges petroleum ether phase, medicinal extract is concentrated under reduced pressure to obtain；
(3) medicinal extract for obtaining step (2), first through depressurizing silica gel column chromatography, using petroleum ether-ethyl acetate mixed solvent conduct
Eluant, eluent carries out gradient elution, and gradient is respectively 100:0,90:10,80:20,70:30,60:40,50:50,40:60,
30:70,20:80,10:90,0:100, each gradient collect two column volumes, are divided into 10 components according to polarity size, wherein
It is component 1 that gradient 100:0~90:10, which is afforded, and it is component 2, gradient 80:20 that gradient 90:10~80:20, which is afforded,
It is component 3 that~70:30, which is afforded, and it is component 4 that gradient 70:30~60:40, which is afforded, and gradient 60:40~50:50 is washed
De- to obtain being component 5, it is component 6 that gradient 50:50~40:60, which is afforded, and gradient 40:60~30:70 is afforded
For component 7, it is component 8 that gradient 30:70~20:80, which is afforded, and it is component 9 that gradient 20:80~10:90, which is afforded,
It is component 10 that gradient 10:90~0:100, which is afforded, and wherein for component 4 first through normal-phase silica gel column chromatography, eluant, eluent is petroleum
Ether: ethyl acetate=15:1-5:1 mixed solvent elutes 2-5 column volume, again through Sephadex LH-20 after reduced pressure
Gel filtration chromatography, eluant, eluent CHCl3: the mixed solvent of MeOH=1:1 elutes 3-6 column volume, again through height after reduced pressure
Effect liquid phase chromatogram HPLC preparation, chromatographic column are Waters C18,9.4 × 250mm, 7 μm, and flow velocity 2mL/min, mobile phase is
MeOH:H2O=55:45 successively obtains compound II, IV, VI, and component 5 is first through Sephadex LH-20 gel filtration chromatography, elution
Agent is MeOH, elutes 3-6 column volume, prepares again through high-efficient liquid phase chromatogram HPLC after reduced pressure, chromatographic column Waters
C18,9.4 × 250mm, 7 μm, flow velocity 2mL/min, mobile phase MeOH:H2O=75:25, successively obtain compound III,
VIII, component 6 are prepared through high-efficient liquid phase chromatogram HPLC, and chromatographic column is Waters C18,9.4 × 250mm, 7 μm, flow velocity 2mL/
Min, mobile phase MeCN:H2O=75:25 successively obtains compound VII, V.
4. method as claimed in claim 3, it is characterised in that it is 75-95% that ethanol solution described in step (1), which is volume fraction,
Ethanol solution, dosage is every kilogram of blue flower radix scutellariae herb 2-3L ethanol solution.
5. a kind of inverase, it is characterised in that the drug is with the described in any item compound of formula I of claim 1-2, its solid
Isomers, tautomer or its pharmaceutically acceptable salt are as effective component.
6. drug described in claim 5, it is characterised in that the drug further includes other antiviral drugs.
7. the described in any item drugs of claim 5-6, it is characterised in that the drug further includes pharmaceutic adjuvant.
8. drug as claimed in claim 7, it is characterised in that pharmaceutic adjuvant is selected from pharmaceutically acceptable carrier, excipient or dilute
Release agent.
9. the described in any item compound of formula I of claim 1-2, its stereoisomer, tautomer or its can pharmaceutically connect
The salt received is preparing the purposes in inverase.
10. purposes as claimed in claim 9, it is characterised in that the inverase is for treating and/or preventing to be felt by HIV-1
Disease caused by contaminating.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710497562.5A CN107141305B (en) | 2017-06-27 | 2017-06-27 | It is a kind of to be isolated from the blue novel Crow alkane type diterpene-kind compound for spending radix scutellariae and its in the purposes prepared in inverase |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710497562.5A CN107141305B (en) | 2017-06-27 | 2017-06-27 | It is a kind of to be isolated from the blue novel Crow alkane type diterpene-kind compound for spending radix scutellariae and its in the purposes prepared in inverase |
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| CN107141305A CN107141305A (en) | 2017-09-08 |
| CN107141305B true CN107141305B (en) | 2018-12-18 |
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| TW201016226A (en) * | 2008-07-14 | 2010-05-01 | Herbalscience Group Llc | Anti-inflammatory and anti-allergy extracts from nettle |
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