CN115894419B - Iridoid compound in viburnum yangbiense leaves as well as preparation method and application thereof - Google Patents
Iridoid compound in viburnum yangbiense leaves as well as preparation method and application thereof Download PDFInfo
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- WGLUMOCWFMKWIL-UHFFFAOYSA-N dichloromethane;methanol Chemical compound OC.ClCCl WGLUMOCWFMKWIL-UHFFFAOYSA-N 0.000 claims description 13
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- 238000000034 method Methods 0.000 claims description 6
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- 235000020357 syrup Nutrition 0.000 claims description 2
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- 229940022698 acetylcholinesterase Drugs 0.000 abstract description 9
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- 208000024827 Alzheimer disease Diseases 0.000 description 8
- HEDRZPFGACZZDS-MICDWDOJSA-N Trichloro(2H)methane Chemical compound [2H]C(Cl)(Cl)Cl HEDRZPFGACZZDS-MICDWDOJSA-N 0.000 description 8
- 239000000243 solution Substances 0.000 description 7
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical group [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 4
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 4
- OIPILFWXSMYKGL-UHFFFAOYSA-N acetylcholine Chemical compound CC(=O)OCC[N+](C)(C)C OIPILFWXSMYKGL-UHFFFAOYSA-N 0.000 description 4
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- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 4
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- ADEBPBSSDYVVLD-UHFFFAOYSA-N donepezil Chemical compound O=C1C=2C=C(OC)C(OC)=CC=2CC1CC(CC1)CCN1CC1=CC=CC=C1 ADEBPBSSDYVVLD-UHFFFAOYSA-N 0.000 description 2
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- KIUMMUBSPKGMOY-UHFFFAOYSA-N 3,3'-Dithiobis(6-nitrobenzoic acid) Chemical compound C1=C([N+]([O-])=O)C(C(=O)O)=CC(SSC=2C=C(C(=CC=2)[N+]([O-])=O)C(O)=O)=C1 KIUMMUBSPKGMOY-UHFFFAOYSA-N 0.000 description 1
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- ZEEBGORNQSEQBE-UHFFFAOYSA-N [2-(3-phenylphenoxy)-6-(trifluoromethyl)pyridin-4-yl]methanamine Chemical compound C1(=CC(=CC=C1)OC1=NC(=CC(=C1)CN)C(F)(F)F)C1=CC=CC=C1 ZEEBGORNQSEQBE-UHFFFAOYSA-N 0.000 description 1
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Classifications
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
An iridoid compound in Viburnum yangbiensis leaves, a preparation method and application thereof, belongs to the technical field of medicines, and relates to 2 novel iridoid compounds (compounds 1-2) extracted and separated from dry leaves of Viburnum yangbiensis (Viburnum chingii Hsu) plants of the genus Viburnum of the family of the five-Fuhuaceae (Adoxaceae). The crude extract of the viburnum yangbiense is separated by a chromatographic method of silica gel column chromatography, namely, the crude extract is rapidly separated into four crude fractions (Fr.1-4), and the four crude fractions are sequentially separated by a chromatographic method of D101 column chromatography, ODS column chromatography, silica gel column chromatography and HPLC separation. And the compounds have inhibitory activity against acetylcholinesterase.
Description
Technical Field
The invention belongs to the technical field of medicines, and particularly relates to an iridoid compound in viburnum sargenti leaves, and a preparation method and application thereof.
Background
The Viburnum yangbiense (Viburnum chingii Hsu) is a plant of the genus Viburnum (Viburn L.) of the family Pacific (Adoxaceae), and is also named meat She Jia and thin-stalk Viburnum yangbiense. To date, literature searches have shown that there is currently only one phytochemical study on this species, and no related report on iridoids and pharmacological activity has been found.
Acetylcholinesterase (AChE) hydrolyzes acetylcholine (ACh) to terminate the pulsatile transmission of cholinergic synapses, present in cholinergic brain synapses and neuromuscular junctions. Alzheimer's Disease (AD) is a neurodegenerative disease that occurs mainly in the elderly. The incidence of AD rises year by year as the population ages. Prevention of AD by increasing the level of acetylcholine in the nervous system is one of the most common methods, meaning that AD can be prevented by lowering AChE levels. The use of acetylcholinesterase inhibitors is one of the most promising methods for increasing brain acetylcholine levels, and can be used in alzheimer's disease and related neurodegenerative diseases.
Disclosure of Invention
The invention aims to provide an iridoid compound in viburnum yangense leaves, a preparation method and application thereof, in particular to an iridoid (1-2) compound separated from leaves of a medicinal plant viburnum yangense (Viburnum chingii Hsu) of the genus viburnum of the family of the five-Fucaceae, an extraction and preparation method of the iridoid compound, and application of the iridoid compound as an acetylcholinesterase inhibitor in the aspect of resisting Alzheimer disease.
The iridoid compound in the viburnum yangbiense leaves or the pharmaceutically acceptable salt of the compound has the structure shown in the following (1) or (2):
Wherein the iridoid compounds shown in the structural formula (1) or (2) are all 3R,4S isomerism.
The preparation method of the iridoid compound in the viburnum yangbiense leaves specifically comprises the following steps:
S1, extracting dry leaves of the viburnum yangense with industrial ethanol with the volume concentration of 70-90% under heating and reflux, combining extracting solutions, and concentrating the extracting solutions to obtain extractum;
s2, suspending the obtained extract with water, extracting with n-butanol for 3-4 times, and removing solvent from the n-butanol layer to obtain dry extract;
s3, subjecting the dry extract to silica gel column chromatography, performing gradient elution by using a dichloromethane-methanol system, and combining the obtained fractions to obtain 4 fractions, namely Fr.1, fr.2, fr.3 and Fr.4; wherein Fr.2 is the sum of the parts by volume of methylene dichloride-methanol with the volume ratio of 40:1, 30:1, 20:1 and 10:1.
S4, carrying out gradient elution on the fraction Fr.2 by an ethanol-water system through a D101 column chromatography, and obtaining 3 fractions after combination of silica gel thin layer detection and HPLC analysis, wherein the 3 fractions are respectively ethanol accounting for 40%,60% and 80% of the volume of the ethanol-water system, and are sequentially recorded as Fr.2.1, fr.2.2 and Fr.2.3;
S5, performing gradient elution on Fr.2.1, fr.2.2 and Fr.2.3 respectively through ODS column chromatography by using an ethanol-water system, and performing cross combination through silica gel thin layer identification and HPLC analysis to obtain 3 fractions, wherein the 3 fractions are respectively 40%,60% and 80% of ethanol in the volume percentage of the ethanol-water system, and are sequentially marked as Fr.A, fr.B and Fr.C;
S6, performing gradient elution on Fr.B through a silica gel column chromatography by using a dichloromethane-methanol system, wherein the obtained fractions are Fr.B1-Fr.B9; fr.b2 is the sum of the methylene chloride-methanol volume ratio of 40:1 and 35:1 fractions.
S7, separating a methanol-water system with the volume ratio of Fr.B2 of 53:47 by utilizing preparative HPLC to obtain 8 components Fr.B2.1-Fr.B2.8; fr.b2.5 is the fraction with HPLC peak time 53-65 min;
s8, eluting the component Fr.B2.5 by utilizing semi-preparative HPLC (high performance liquid chromatography) in an acetonitrile-water system with the volume ratio of 20:80, wherein the flow rate is 2.5ml/min, and the compound 1 is obtained at 32min and the compound 2 is obtained at 34 min;
In the preparation method, the dry Viburnum yangensis leaves are obtained from Viburnum yangensis (Viburn L.) plants of the genus Viburnum of the family Pachyrhizus (Adoxaceae) of the family Pachyrhizus yangensis (Viburnum chingii Hsu).
In the preparation method, in S1, the dry leaves of the viburnum sargenti are extracted for 2-3 times by reflux with 70-90% industrial ethanol;
In the preparation method, in S1, the extract is concentrated under reduced pressure to obtain extract, and the temperature of the reduced pressure concentration is 55-65 ℃.
In the preparation method, in S3, the gradient elution of a methylene dichloride-methanol system is carried out, and the volume change is 100:1-1:1.
In the preparation method, in S4, the fraction Fr.2 is subjected to gradient elution by a D101 ethanol-water system with the volume ratio of 20:80-90:10, and the volume concentration of the adopted ethanol is 20-90%;
In the preparation method, in S5, fractions Fr.2.1, fr.2.2 and Fr.2.3 are subjected to gradient elution by an ODS column chromatography in a volume ratio of 20:80-90:10 ethanol-water system, and the volume concentration of the adopted ethanol is 20% -90%;
In the preparation method, in S6, the fraction Fr.B is subjected to gradient elution by a silica gel column chromatography in a dichloromethane-methanol system with the volume ratio of 45:1-7:1;
in the preparation method, in S7, the fraction Fr.B2 is subjected to isocratic elution by a methanol-water system with the volume ratio of 53:47 through preparation HPLC;
In the preparation method, in S8, the fraction Fr.B2.5 is subjected to isocratic elution by semi-preparative HPLC in an acetonitrile-water system with a volume ratio of 20:80.
The iridoid compound (compound 1-2) has the activity of inhibiting acetylcholinesterase, and the result shows that the compound 1,2 has the activity of inhibiting acetylcholinesterase, and the IC 50 value is 1.284+/-0.277 to 2.090 +/-0.095 mu M.
A pharmaceutical composition comprises iridoid compound in the Viburnum yangbiense leaves, or pharmaceutically acceptable salt of the compound, and one or combination of pharmaceutically acceptable carrier, excipient and diluent; the drug composition is orally taken or injected, and the dosage form comprises: tablets, capsules, powders, syrups and injections.
The invention relates to an application of iridoid compounds or pharmaceutical compositions in viburnum yangbiense leaves in preparation of acetylcholinesterase inhibitors.
The iridoid compound or the pharmaceutical composition in the viburnum yangbiense leaves is used for preparing the anti-Alzheimer disease medicine.
The invention has the beneficial effects that:
The invention provides a novel iridoid compound for the field of Alzheimer disease resistance, and the compounds are easy to separate and purify, are all optical pure compounds with determined stereo configuration, and have further development value. In addition, the method has the characteristics of reasonable design, simple process, low cost and industrial production.
Drawings
Hresis spectrum of compound 1 of fig. 1;
the UV spectrum of compound 1 of fig. 2;
FIG. 3 1 H NMR spectrum of Compound 1 (600 MHz, DMSO-d 6);
FIG. 4 13 C NMR spectrum of Compound 1 (150 MHz, DMSO-d 6);
FIG. 5 HSQC spectrum of Compound 1 (600 MHz, DMSO-d 6);
FIG. 6 HMBC spectra (600 MHz, DMSO-d 6) of Compound 1;
FIG. 7 NOESY spectrum of Compound 1 (600 MHz, DMSO-d 6);
FIG. 8 1H-1 H COSY spectrum of Compound 1 (600 MHz, DMSO-d 6);
FIG. 9 critical HMBC of Compounds 1-2;
FIG. 10 key NOE correlation for Compounds 1-2;
FIG. 11 HRESIMS spectrum of Compound 2;
FIG. 12 UV spectrum of Compound 2;
FIG. 13 1 H NMR spectrum of Compound 2 (600 MHz, DMSO-d 6);
FIG. 14 13 C NMR spectrum of Compound 2 (150 MHz, DMSO-d 6);
FIG. 15 HSQC spectrum of Compound 2 (600 MHz, DMSO-d 6);
FIG. 16 HMBC spectra (600 MHz, DMSO-d 6) of Compound 2;
FIG. 17 NOESY spectrum of Compound 1 (600 MHz, DMSO-d 6);
FIG. 18 1H-1 H COSY spectrum of Compound 1 (600 MHz, DMSO-d 6);
FIG. 19 shows a graph of the comparison of measured ECD to calculated ECD for Compounds 1-2; wherein (a) is compound 1; (b) is compound 2;
FIG. 20 binding interactions of Compounds 1,2 with acetylcholinesterase.
Detailed Description
The present invention will be described in further detail with reference to examples.
Example 1
The preparation of the compound iridoid (1-2) comprises the following steps:
Reflux-extracting dry leaves of the viburnum yangbiense with 70% -90% industrial ethanol for 2 times, mixing the extracting solutions, concentrating under reduced pressure at 60 ℃ to obtain an extract, suspending the obtained extract with water, extracting with n-butanol for 3-4 times, subjecting the obtained dry extract after removing the solvent from the n-butanol layer to silica gel column chromatography, carrying out gradient elution (100:1-1:1, v:v) with a dichloromethane-methanol system, and carrying out cross-combination on the obtained fractions to obtain 4 fractions Fr.1 (132 g) (the volume ratio of dichloromethane-methanol is the sum of 100:1 and 50:1), fr.2 (150 g) (the volume ratio of dichloromethane-methanol is the sum of 40:1, 30:1, 20:1 and 10:1), fr.3 (100 g) (the volume ratio of dichloromethane-methanol is the sum of 8:1, 5:1 and 2:1), fr.4 (200 g) (the volume ratio of dichloromethane-methanol is 1:1);
Subjecting the fraction Fr.2 to D101 column chromatography, gradient eluting with ethanol-water as eluent (20:80-90:10, v: v), and combining by silica gel thin layer detection and HPLC analysis to obtain 3 fractions Fr.2.1 (30 g) (ethanol-water volume ratio 40:60), fr.2.2 (50 g) (ethanol-water volume ratio 60:40), fr.2.3 (50 g) (ethanol-water volume ratio 80:20);
Fr.2.1, fr.2.2 and Fr.2.3 are respectively subjected to gradient elution (20:80-90:10, v: v) by an ODS column chromatography in an ethanol-water system, and 3 fractions Fr.A (14.5 g) (the ethanol-water volume ratio is 40:60 fractions), fr.B (12.5 g) (the ethanol-water volume ratio is 60:40) and Fr.C (20.4 g) (the ethanol-water volume ratio is 80:20 fractions) are obtained by cross-combining through silica gel thin layer identification and HPLC analysis;
performing gradient elution on Fr.B through a silica gel column chromatography with a volume ratio of 45:1-7:1 methylene dichloride-methanol system to obtain 9 fractions Fr.B1-Fr.B9; wherein Fr.B2 (2 g) is a fraction obtained by mixing methylene dichloride and methanol in a volume ratio of 40:1 to 35:1;
Performing isocratic separation on Fr.B2 by using a methanol-water system with the volume ratio of 53:47 to obtain 8 components Fr.B2.1-Fr.B2.8; wherein Fr.B2.5 (15 mg) is the fraction with the HPLC peak time of 53-65 min;
Component fr.b.2.5 was subjected to isocratic separation by semi-preparative HPLC in an acetonitrile-water system at a volume ratio of 20:80, at a flow rate of 2.5ml/min, to give compound 1 (3 mg) at 32min, and compound 2 (4 mg) at 34 min.
The structural identification result of the compound obtained by the invention is as follows:
the structure and relative configuration of compounds 1-2 were determined using ultraviolet spectroscopy, high resolution mass spectrometry, one-dimensional and two-dimensional NMR techniques. The absolute configuration of the measured ECD is determined by comparing the measured ECD with the calculated ECD spectra (see FIGS. 1-19).
Compound 1:
compound 1 is white needle crystal (methanol), is easy to dissolve in solvents such as methanol, dichloromethane and the like, Molecular ion peaks [ M+Na ] +m/z 251.0892(calcd for C11H16O5 Na,251.0890 were given by HRESIMS, and the molecular formula was determined to be C 11H16O5, unsaturation 4 (FIG. 1). UV (CH 3OH)λmax (log epsilon): 252nm (1.183) (fig. 2). 1H-NMR(600MHz,CDCl3) spectrum (fig. 3), the high field region gives 2 methyl proton signals δ H 2.20 (3 h, d, j=1.4), including one methoxy proton signal δ H 3.69.69 (3 h, d, j=1.35), 2 oxygen methine signals δ H 3.58.58 (1 h, dd, j= 10.8,6.5), 3.47 (1 h, m), 3.92 (1 h, dd, j= 11.5,2.6), 4.36 (1 h, m), proton signals δ H7.58(1H,s).13C-NMR(150MHz,CDCl3 on a set of double bonds show 11 carbon signals in spectrum (fig. 4), δ C 196.1,172.6 is 2 hydroxy carbon signals, δ C 63.3,61.9 is 2 oxygen carbon signals, two methyl carbon signals δ C 25.1.1, including one methoxy carbon signal δ C 51.9.9, and a set of double bond carbon signals δ C 157.8,118.03.
The HMBC spectra (FIG. 6) combined with the HSQC spectra (FIG. 5) gave delta H.58 (H-1) correlated with delta C.26.7 (C-3), 63.3 (C-5), 196.1 (C-6) and 118.0 (C-2) presence, delta H 2.00 (H-4) correlated with delta C.7 (C-3), 172.6 (C-9), 37.9 (C-4) and 118.0 (C-2) presence, delta H 4.36,3.92(H2 -5) correlated with delta C.7 (C-2), 61.9 (C-10), 157.8 (C-1) presence, delta H 4.36,3.92(H2 -5) correlated with delta C.26.7 (C-2), 61.9 (C-10), 157.8 (C-1) presence, and delta H.7.58 (H-1) correlated with delta 3463.3 (C-5) presence, thereby determining the six-membered ring composition of C-1, C-3, C-4, C-5. Delta H 2.78,2.19(H2 -8) correlated with delta C 26.7.7 (C-3), 36.9 (C-4), 118.0 (C-2), 172.6 (C-9) presence, and delta H 3.69(-OCH3) correlated with delta C 172.6.6 (C-9), indicating that-OCH 3 is attached to C-9, determining the presence of C-3 side chain segments, delta H 2.20 (H-7) correlated with delta C 157.8 (C-1) and 196.1 (C-6) presence, indicating that-CH 3 is attached to C-6, determining the presence of C-2 side chain segments. The key HMBC signal is shown in fig. 9, 1H-1 H COSY spectrum further confirms the structural fragment (fig. 8), whereby the planar structure of compound 1 was determined.
The relative configuration of compound 1 was determined by NOESY spectra (FIG. 7), NOSEY spectra, delta H 2.96 (H-3) and delta H3.58,3.47(H2 -10) correlated, indicating that H-3 and H-10 (FIG. 10) are on the same side, and thus the relative configuration of compound 1 was 3R, 4S.
The absolute configuration of compound 1 was determined by comparing the calculated and measured ECD. The ECD curve measured for Compound 1 was better matched with the ECD curve calculated for 3R,4S configuration (FIG. 19 (a)), thus determining the absolute configuration of Compound 1 as 3R,4S. In conclusion, the structure of the compound 1 is determined, hydrocarbon signals of the compound are attributed, and the compound is determined to be a novel compound which is not reported after literature retrieval.
Compound 2:
the compound 2 is white needle crystal (methanol), is easy to dissolve in solvents such as methanol, dichloromethane and the like, Molecular ion peaks [ M+Na ] +m/z 251.0894(calcd for C11H16O5 Na,251.0890 were given by HRESIMS, and the molecular formula was determined to be C 11H16O5, unsaturation 4 (FIG. 11). UV (CH 3OH)λmax (log epsilon): 263nm (1.457), 205nm (1.526) (fig. 12). 1H-NMR(600MHz,CDCl3) spectrum (fig. 13), the high field region gives 2 methyl proton signals δ H 2.21 (3 h, d, j=1.1), including one methoxy proton signal δ H 3.69.69 (3 h, d, j=2.7), 2 even oxymethylene signals δ H 3.58.58 (1 h, dd, j= 10.7,6.3), 3.47 (1 h, m), 2.79 (1 h, dd, j= 16.2,3.3), 2.18 (1 h, m), one methylene proton signal δ H 4.38(1H,dt,J＝11.5,1.9),4.00(1H,dd,J＝11.5,2.8).13C-NMR(150MHz,CDCl3) spectrum (fig. 14) shows 11 carbon signals, δ C 189.3,172.6 is 2 hydroxy carbon signals, δ C 63.7,61.9 is 2 even oxygen carbon signals, two methyl carbon signals δ C 16.7, including one methoxy carbon signal δ C 51.8.8, and a set of double bond carbon signals δ C 171.6,114.5.
HMBC spectra (FIG. 16) combined with HSQC spectra (FIG. 15) showed that delta H 2.21(CH3 -7 was correlated with delta C 26.4.4 (C-3), 114.5 (C-2), 171.6 (C-1) and 37.9 (C-4) presence, delta H 1.99 (H-4) was correlated with delta C 26.4.4 (C-3), 61.9 (C-10) and 144.5 (C-2) presence, delta H 2.79.79 (H-8) was correlated with delta C 26.4.4 (C-3), 37.9 (C-4), 114.5 (C-2) and 172.6 (C-9) presence, delta H 9.81.81 (H-6) was correlated with delta C 26.4.4 (C-3) and 114.5 (C-2) presence, and delta H 4.38.38 (H-5) was correlated with delta C 26.4.4 (C-3), 171.6 (C-1) presence, thereby determining the six-membered ring consisting of C1, C2, C3, C4-membered ring. Delta H 2.21(CH3 -7) correlated with delta C 171.6.6 (C-1), 114.5 (C-2), indicating that-CH 3 was linked to C-1, delta H3.58,3.47(H2 -10) correlated with delta C 26.4.4 (C-3), 63.7 (C-5) and 37.9 (C-4), indicating that-CH 2 OH was linked to C-4, determining the presence of a C-4 side chain fragment, delta H 3.69(-OCH3) correlated with delta C 172.6 (C-9), delta H 2.94.94 (H-3) correlated with delta C 38.6.6 (C-8), 172.6 (C-9), determining the presence of a C-3 side chain fragment, the critical HMBC signal is shown in FIG. 9, and the 1H-1 H COSY spectrum further confirms the structural fragment (FIG. 18), whereby the compound 2 planar structure was determined.
The relative configuration of compound 2 was determined by NOESY spectra (FIG. 17), NOSEY spectra, delta H 2.94.94 (H-3) and delta H3.58,3.47(H2 -10) correlated, indicating that H-3 and H-10 are on the same side (FIG. 10), and thus the relative configuration of compound 2 was 3R, 4S. Its absolute configuration was determined to be 3R,4S by comparing the measured ECD with the calculated ECD (FIG. 19 (b)).
In conclusion, the structure of the compound 2 is determined, hydrocarbon signals of the compound are attributed, and related reports are not searched after the literature is consulted, so that the compound is determined to be a new compound.
TABLE 1 NMR data for Compounds 1-2 1 H (600 MHz) and 13 C (150 MHz) in DMSO-d 6
Example 2
Acetylcholinesterase inhibitory Activity
Samples were diluted to the appropriate concentration (100,40,20,10,1. Mu. Mol/L) using PBS, 50. Mu.L of PBS solution, 25. Mu.L of sample solution, 12.5. Mu.L of enzyme solution, 125. Mu.L of DTNB solution were added to wells of a 96-well plate in advance, mixed and then cooled overnight, then 50. Mu.L of ATCI solution was added, absorbance was measured at 412nm immediately using a microplate reader, and 5 minutes later, donepezil was used as a positive control.
The results showed that compounds 1,2 showed inhibitory activity against acetylcholinesterase (see FIG. 20), with IC 50 values of 1.284.+ -. 0.277 to 2.090.+ -. 0.095. Mu.M.
Claims (9)
1. An iridoid compound represented by the following structural formula (1) or (2), or a pharmaceutically acceptable salt thereof:
2. The iridoid compound according to claim 1, characterized in that: wherein the iridoid compound shown in the structural formula (1) or (2) is 3R,4S isomerism.
3. The iridoid compound according to claim 1, characterized in that: the iridoid compound is extracted and separated from viburnum sargenti (Viburnum chingii) belonging to genus viburnum of family Pavetaceae.
4. The method for producing an iridoid compound according to claim 1, characterized in that: the method comprises the following steps:
(1) Reflux-extracting dry leaves of the viburnum yangbiense with ethanol and concentrating to obtain extract;
(2) Extracting the extract with n-butanol and removing solvent to obtain dry extract;
(3) Separating the dry extract into four crude fractions Fr.1-Fr.4 by vacuum silica gel column chromatography; wherein Fr.2 is the sum of the parts by volume of methylene dichloride and methanol with the volume ratio of 40:1-10:1;
(4) Separating Fr.2 by D101 and reversed-phase ODS column chromatography to obtain Fr.A-Fr.C; wherein Fr.B is a fraction of 60% of ethanol in the reversed-phase ODS column chromatography separation by volume percent of the ethanol-water system;
(5) Separating Fr.B by silica gel column chromatography to obtain 9 fractions Fr.B1-Fr.B9; wherein Fr.B2 is the sum of the obtained flow parts with the volume ratio of dichloromethane to methanol being 40:1-35:1;
(6) Separating Fr.B2 by preparative high performance liquid chromatography to obtain Fr.B2.1-Fr.B2.8; wherein Fr.B2.5 is a fraction with the peak time of 53-65min in the preparation of high performance liquid chromatography;
(7) Fr.B2.5 is separated by semi-preparative high performance liquid chromatography to obtain compound 1 of structural formula (1) and compound 2 of structural formula (2).
5. The method for producing an iridoid compound according to claim 4, characterized in that: the volume percentage concentration of the ethanol in the step (1) is 70-90 percent; the extraction times are 2-3 times; and/or, the temperature of the reduced pressure concentration in the step (2) is 55-65 ℃; and/or, the gradient elution condition of the reduced pressure silica gel column chromatography in the step (3) is CH 2Cl2:meoh=100:1 to 1:1.
6. The method for producing an iridoid compound according to claim 4, characterized in that: gradient elution conditions of the D101 column chromatography in the step (4) are EtOH 2 O=20:80-90:10; and/or gradient elution conditions for reversed-phase ODS column chromatography are EtOH 2 o=20:80-90:10.
7. The method for producing an iridoid compound according to claim 4, characterized in that: the silica gel column chromatography elution condition in the step (5) is dichloromethane-methanol=45:1-7:1; and/or, the preparative HPLC conditions in step (6) are methanol-water=53:47; and/or, the semi-preparative HPLC condition in step (7) is acetonitrile-water=20:80.
8. A pharmaceutical composition comprising the iridoid compound of claim 1 or 2, or a pharmaceutically acceptable salt thereof, and further comprising one or a combination of a pharmaceutically acceptable carrier, excipient, diluent; the drug composition is orally taken or injected, and the dosage form comprises: tablets, capsules, powders, syrups and injections.
9. Use of an iridoid compound according to claim 1 or 2 or a pharmaceutical composition according to claim 8 for the preparation of an anti-alzheimer's disease medicament.
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| CN108774276A (en) * | 2018-08-07 | 2018-11-09 | 扬州大学 | Radix Viburni fordiae fruit lignans extract and its active constituent and purposes |
| CN114195641A (en) * | 2020-09-18 | 2022-03-18 | 沈阳药科大学 | Seven-element vibsan diterpenoid compound and preparation and application thereof |
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| CN108774276A (en) * | 2018-08-07 | 2018-11-09 | 扬州大学 | Radix Viburni fordiae fruit lignans extract and its active constituent and purposes |
| CN114195641A (en) * | 2020-09-18 | 2022-03-18 | 沈阳药科大学 | Seven-element vibsan diterpenoid compound and preparation and application thereof |
Non-Patent Citations (2)
| Title |
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| Two new iridoids and triterpenoid analogues from the leaves of Viburnum chingii and their anti-acetylcholinesterase activity;Jing-Jing Liang等;《Fitoterapia》;第1-8页 * |
| 暴马子有效成分研究;梁文藻等;《药学学报》;19861231;第21卷(第12期);第906-911页 * |
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