CN1315866C - Radix asparagi steroid saponin extract and its preparing process and application - Google Patents
Radix asparagi steroid saponin extract and its preparing process and application Download PDFInfo
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Abstract
The present invention provides total steroid saponin extract of radix asparagi, a preparation method and the application thereof. Six compounds are separated out of steroid saponin extract that is prepared, and four of the compounds are identified as novel compounds. The preparation method comprises the way of n-butyl alcohol extraction and the way of macroporous resin adsorption. Proved by the research of pharmacological activity, the prepared total steroid saponin extract of radix asparagi, which has the activity of cardiac vessels and cerebral vessels, can obviously increase the cerebral blood flow of anesthetized dogs, obviously reduce the cerebral vascular resistance of anesthetized dogs, and produce unobvious influence on the mean arterial pressure and heart rate of anesthetized dogs. Therefore, the total steroid saponin extract of radix asparagi can be used for preparing medicines or food for preventing and curing cardiac diseases and cerebral diseases.
Description
Technical field
Effective component extracts that the present invention relates to obtain from natural drug and preparation method thereof and application are the total steroid saponin extract that obtains about by the Chinese medicine asparagus fern specifically, and its preparation method reaches in preparation cardiovascular medicament or Application in Food.
Background technology
The Chinese medicine asparagus fern is the piece root of liliaceous plant Radix Asparagi [Asparagus cochinchinensis (Lour.) Merr.], have nourshing Yin and drynsessmoistening prescription, effect of fire falls in clearing lung-heat, cure mainly cough caused by dryness-heat, consumption of YIN caused by febrile disease, interior heat is quenched one's thirst, the dry constipation of intestines, swelling and pain in the throat etc. [editorial committee of State Administration of Traditional Chinese Medicine's " China's book on Chinese herbal medicine " compiles. and " China's book on Chinese herbal medicine "; Shanghai science tech publishing house, 1999.8 (22): 63～69].Asparagus fern is a conventional Chinese medicine, and aboundresources.TenjiK etc. [Tenji K, et al.Chem Pharm Bull, 1979,27 (12): 3086] from asparagus fern isolation identification 4 asparagus fern furan sterol oligosaccharides glycosides (ASPIV; ASPV; ASPVI; ASPVII); [Liang Z Z such as Liang Z Z, et al.Planta Med, 1988,54 (4): 344] from asparagus fern isolation identification Smilax saponin B (Methylprotodiosin), pseudo-protodioscin (Pseudoprotodiosin) and 3-O-[α-L-rhamnopyranosyl (1 → 4)-β-D-glucopyranosyl]-26-O-(β-D-glucopyranosyl)-(25R)-5,20-furan steroid diene-3 β-, the 26-glycol 3-O-[α-L-rhamanopyranosyl (1 → 4)-β-D-glucopyranosyl]-26-O-(β-D-glucopyranosyl)-(25R)-furosta-5,20-dien-3,20-diol}; [Masashi T, etal.Chem Pharm Bull, 1974,22 (10): 2306] such as Masashi T. have isolated 7 oligosaccharides and sucrose from asparagus fern; Holt just wait [holt just, etc. study of pharmacy (day), 1958,30:477] from asparagus fern, got glucose, fructose, 5-methoxymethyl furfural, β-Gu Zaichun; Masashi T. etc. [Masashi T, et al.CA, 1976,85:30677y] and Ni Jingman etc. [Ni Jingman, etc. herbal medicine, 1992,23 (4): 182] reported 19 seed amino acid compositions; Du Xuhua etc. [Du Xuhua etc. Shenyang Pharmacy College's journal, 1990,7 (3): 197] from asparagus fern, told 4 kinds of asparagus fern polysaccharide; Open the prosperous grade of portion [portion of opening is prosperous etc. the research University of Anhui journal (natural science edition) of the separation and purification of Radix Asparagi glycoprotein and cardohydrata-peptide linkage, 1996,20 (4): 51～54] reported the separation and purification and the structural research of asparagus fern glycoprotein.So far do not see that the compound that above-mentioned separation obtains has the active report of cardiovascular and cerebrovascular.
Summary of the invention
The invention provides a kind of asparagus fern total steroid saponin extract and preparation method thereof and application, separate obtaining 6 kinds of compounds from the total steroid saponin extract, their chemical structure is identified as follows:
Compound (1): (25S)-26-O-β-D-glucopyanosyl base-5 β-furan steroid-20 (22)-alkene-3 β, 26-glycol-3-O-α-L-sandlwood pyrans glycosyl (1 → 4)]-β-D-glucopyranoside (25S)-and 26-O-β-D-glucopyranosyl-5 β-furosta-20 (22)-en-3 β, 26-diol-3-O-[α-L-rhamnopyranosyl (1 → 4)]-β-D-glucopyranoside}.Physico-chemical property and spectroscopic data are as follows: white amorphous powder (MeOH).mp：189-191℃。[α]
D 20-33.06°(C＝4.84，MeOH)。The Liebermann-Burchard reaction is positive, and the Ehrlich reagent react takes on a red color, and illustrates that this compound is the steroidal saponin of F ring open loop.ESI-MS shows: 909[M+Na]
+, point out this compound molecular weight to should be 886; Molecular formula is C
45H
74O
17 1H-NMR and
13C-NMR sees Table 1 and table 2.
Table 1 compound (1)
1H-NMR (C
5D
5N, 400MHz)
| NO. | δ H/ppm | NO. | δ H/ppm |
| 17-H CH 3-18 CH 3-19 CH 3-21 CH 3-27 | 2.49(1H，d，J＝10.1) 0.69(3H，s) 0.85(3H，s) 1.63(3H，s，) 1.05(3H，d，J＝6.0) | 26-Glu 1 6 3-Glu 1 6 4’-Rha 1 6 | 4.81(1H，d，J＝7.7) 4.39(1H，brd，J＝124) 4.57(1H，brd，J＝124) 4.83(1H，d，J＝7.8) 4.12(1H，brd，J＝10.3) 4.26(1H，brd，J＝10.3) 5.9(1H，brs) 1.7(3H，d，J＝6.2) |
Table 2 compound (1)
13C-NMR (C
5D
5N, 400MHz)
| NO. | δ C/ppm | NO. | δ C/ppm |
| 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 | 30.6 26.5 74.2 30.2 36.6 26.6 26.6 34.9 39.8 34.9 21.0 39.8 43.5 54.4 31.1 84.2 64.3 14.1 23.5 103.2 11.5 152.1 34.1 23.3 33.4 74.9 16.8 | 26-Glu 1 2 3 4 5 6 3-Glu 1 2 3 4 5 6 4’-Rha 1 2 3 4 5 6 | 104.8 75.2 78.2 72.5 78.1 62.5 102.7 74.9 76.4 78.01 76.8 61.8 102.3 72.3 72.5 73.7 69.8 18.2 |
Compound (2): Sarsasapogenin-3-O-[α-L-sandlwood pyrans glycosyl (1 → 4)]-β-D-glucopyranoside 3-O-[α-L-rhamnopyranosyl (1 → 4)]-β-D-glucopyranoside-(25S)-5 β-spirostan-3 beta-ol }.Physico-chemical property and spectroscopic data are as follows: colourless needle crystal (CHCl
3).The Liebermann-Burchard reaction is positive.mp：252-254℃。[α]
D 20-61.5°[C＝0.80，CHCl
3/MeOH(1∶1)]。ESI-MS provides [M+Na]
+Peak 747 infers that compound molecular weight is 724; Molecular formula is C
39H
64O
12NMR sees Table 3.With document [the black positive allusion quotation of willow etc., foreign medical science, traditional Chinese medicine fascicle, 1988; 10 (1): 56] unanimity.
Table 3 compound (2)
1H-NMR (DMSO-D6,400MHz)
| NO. | δ H(J Hz) | δ C | NO. | δ H(J Hz) | δ C |
| 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 | 0.9(3H，s) 0.7(3H，s) 1.02(3H， d，J＝7.06) | 29.3 26.1 73.7 30.0 35.8 26.3 25.9 34.8 39.6 34.6 20.4 39.6 40.1 55.6 31.4 80.3 61.8 16.1 23.5 41.5 14.4 | 22 23 24 25 26 27 3-Glu 1 2 3 4 5 6 Rha 1 2 3 4 5 6 | 3.35(1H，d，J＝9.3)， 4.09(1H，dd，J＝5.3， 10.6) 0.92(3H，d，J＝6.9) 4.19(1H，d，J＝7.8) 4.71(1H，brs) 1.11(3H，d，J＝6.16) | 108.8 25.5 25.4 26.4 64.9 15.9 100.5 72.6 75.3 76.8 75.3 60.0 100.8 70.6 70.7 71.9 68.6 17.7 |
Compound (3): (25R)-26-O-β-D-glucopyanosyl base-furan steroid-5,20-diene-3 β, 26-glycol-3-O-[α-L-sandlwood pyrans glycosyl (1 → 2)]-[(β-D-glucopyanosyl-(1 → 4))-α-L-sandlwood pyrans glycosyl-(1 → 4)]-β-D-glucopyranoside (25R)-26-O-β-D-glucopyranosyl-furosta-5,20-diene-3 β, 26-diol-3-O-[α-L-rhamnopyranosyl (1 → 2)]-[(β-D-glucopyranoside-(1 → 4)-α-L-rhamnopyranosyl-(1 → 4))-β-D-glucopyranoside].Physico-chemical property and spectroscopic data are as follows: white amorphous powder (MeOH).mp：190-192℃。The Liebermann-Burchard reaction is positive, and the Ehrlich reagent react takes on a red color, and illustrates that this compound is the steroidal saponin of F ring open loop.ESI-MS shows: 1215[M+Na]
+, point out this compound molecular weight to should be 1192.Molecular formula is C
57H
92O
26 1H-NMR and
13C-NMR sees Table 4 and table 5.
Table 4 compound (3)
1H-NMR (C
5D
5N, 400MHz)
| NO. | δ H/ppm | NO. | δ H/ppm |
| 17-H CH 3-18 CH 3-19 CH 3-21 CH 3-27 26-Glu 1 6 | 2.45(1H，d，J＝10.1) 0.72(3H，s) 1.04(3H，s) 1.61(3H，s，) 1.02(3H，d，J＝6.7) 4.82(1H，d，J＝7.7) 4.34(1H，brd，J＝12.4) 4.41(1H，br d，J＝12.4) | 3-Glu 1 6 2’-Rha 1 6 4’-Rha 1 6 4”-Glu 1 6 | 4.92(1H，d，J＝7.0) 4.05(1H，brd，J＝10.5) 4.20(1H，brd，J＝10.5) 5.83(1H，brs) 1.75(3H，d，J＝6.2) 6.35(1H，brs) 1.65(3H，d，J＝6.2) 5.21(1H，d，J＝7.7) 4.32(1H，brd，J＝12.4) 4.41(1H，brd，J＝12.4) |
Table 5 compound (3)
13C-NMR (C
5D
5N, 400MHz)
| NO. | δ C/ppm | NO. | δ C/ppm |
| 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 | 37.2 29.8 77.73 38.6 142.1 121.5 32.1 31.1 50.0 36.8 20.9 39.3 43.1 54.6 31.1 84.1 64.1 13.8 19.1 103.2 11.4 152.1 34.1 23.3 33.4 74.2 16.8 | 26-Glu 1 2 3 4 5 6 3-Glu 1 2 3 4 5 6 2’-Rha 1 2 3 4 5 6 4’-Rha 1 2 3 4 5 6 4”-Glu 1 2 3 4 5 6 | 104.8 78.24 74.86 72.13 78.2 62.15 100.1 77.45 73.78 77.59 76.64 60.93 101.64 71.40 72.48 73.78 69.15 18.26 101.69 71.59 72.17 84.97 68.25 17.98 106.45 76.2 78.12 71.01 76.6 62.53 |
Compound (4): (25S)-26-O-β-D-glucopyanosyl base-5 β-furan steroid-20 (22)-alkene-3 β, 15,26-triol-3-O-α-L-sandlwood pyrans glycosyl (1 → 4)]-β-D-glucopyranoside (25S)-26-O-β-D-glucopyranosyl-5 β-furosta-20 (22)-en-3 β, 15,26-triol-3-O-[α-L-rhamnopyranosyl (1 → 4)]-β-D-glucopyranoside}.Physico-chemical property and spectroscopic data are as follows: white amorphous powder (MeOH).mp：210-212℃。The Liebermann-Burchard reaction is positive, and the Ehrlich reagent react takes on a red color, and illustrates that this compound is the steroidal saponin of F ring open loop.ESI-MS shows: 925[M+Na]
+, point out this compound molecular weight should be: 902.Molecular formula is C
45H
74O
18 1H-NMR and
13C-NMR sees Table 6 and table 7.
Table 6 compound (4)
1H-NMR (C
5D
5N, 400MHz)
| NO. | δ H/ppm | NO. | δ H/ppm |
| 17-H CH 3-18 CH 3-19 CH 3-21 CH 3-27 | 2.5(1H，d，J＝10.3) 0.7(3H，s) 0.85(3H，s) 1.72(3H，s，) 1.18(3H，d，J＝6.7) | 26-Glu 1 6 3-Glu 1 6 4’-Rha 1 6 | 4.82(1H，d，J＝7.8Hz) 4.38(1H，brd，J＝12.3) 4.56(1H，brd，J＝12.3) 4.82(1H，d，J＝7.8) 4.14(1H，brd，J＝10.5) 4.27(1H，brd，J＝10.5) 5.9(1H，brs) 1.7(3H，d，J＝6.2) |
Table 7 compound (4)
13C-NMR (C
5D
5N, 400MHz)
| NO. | δ C/ppm | NO. | δ C/ppm |
| 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 | 30.601 26.642 74.894 29.292 36.647 25.516 26.642 34.808 39.845 34.884 20.984 39.736 43.567 54.42 84.37 90.094 64.64 14.182 23.538 104.96 11.36 153.79 34.109 39.421 30.713 75.368 17.697 | 26-Glu 1 2 3 4 5 6 3-Glu 1 2 3 4 5 6 4’-Rha 1 2 3 4 5 6 | 104.86 74.948 78.130 71.384 76.473 62.515 102.69 75.210 78.227 78.084 76.825 61.272 102.36 72.297 72.489 73.696 70.003 18.188 |
Compound (5): (25R)-26-O-β-D-glucopyanosyl base-5 β-furan steroid-20 (22)-alkene-3 β, 26-glycol-3-O-[β-D-glucopyanosyl base-(1 → 2)]-β-D-glucopyranoside (25R)-and 26-O-β-D-glucopyranosyl-5 β-furost-20 (22)-en-3 β, 26-diol-3-O-[β-D-glucopyranosyl-(1 → 2)]-β-D-glucopyranoside}.Physico-chemical property and spectroscopic data are as follows: white amorphous powder (MeOH).mp：188-190℃。The Liebermann-Burchard reaction is positive, and the Ehrlich reagent react takes on a red color, and illustrates that this compound is the steroidal saponin of F ring open loop.ESI-MS shows: 925[M+Na]
+, point out this compound molecular weight should be: 902.Molecular formula is C
45H
74O
18 1H-NMR and
13C-NMR sees Table 8 and table 9.
Table 8 compound (5)
1H-NMR (C
5D
5N, 400MHz)
| NO. | δ H/ppm | NO. | δ H/ppm |
| 17-H CH 3-18 CH 3-19 CH 3-21 CH 3-27 | 2.45(1H，d，J＝10.0) 0.72(3H，s) 1.02(3H，s) 1.65(3H，s，) 1.05(3H，d，J＝5) | 26-Glu 1 6 3-Glu 1 6 2’-Glu 1 6 | 4.80(1H，d，J＝7.0) 4.34(1H，brd，J＝12.5) 4.42(1H，brd，J＝12.5) 4.95(1H，d，J＝7.0) 4.36(1H，brd，J＝10.6) 4.49(1H，brd，J＝10.6) 5.4(1H，d，J＝7.0) 4.37(1H，brd，J＝12.3) 4.5(1H，brd，J＝12.3) |
Table 9 compound (5)
13C-NMR (C
5D
5N, 400MHz)
| NO. | δ C/ppm | NO. | δ C/ppm |
| 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 | 30.656 26.557 74.90 30.488 36.575 26.595 26.690 34.944 39.940 34.944 21.04 39.835 43.58 54.50 31.116 84.29 64.43 14.10 23.17 103.25 11.48 152.12 34.124 23.337 33.396 74.90 16.87 | 26-Glu 1 2 3 4 5 6 3-Glu 1 2 3 4 5 6 2’-Glu 1 2 3 4 5 6 | 104.75 76.60 78.07 71.46 78.24 62.42 101.56 82.76 75.01 71.34 77.89 62.58 105.52 74.86 77.62 71.59 78.12 62.67 |
Compound (6): (25R)-26-O-β-D-glucopyanosyl base-furan steroid-5,20-diene-3 β, 26-glycol-3-O-[α-L-sandlwood pyrans glycosyl (1 → 2)]-[α-L-sandlwood pyrans glycosyl-(1 → 4)]-β-D-glucopyranoside (25R)-26-O-β-D-glucopyranosyl-furost-5,20-diene-3 β, 26-diol-3-O-[α-L-rhamnopyranosyl (1 → 2)]-[α-L-rhamnopyranosyl-(1 → 4)]-β-D-glucopyranoside}.Physico-chemical property and spectroscopic data are as follows: white amorphous powder (MeOH).mp：175-177℃。The Liebermann-Burchard reaction is positive, and the Ehrlich reagent react takes on a red color, and illustrates that this compound is the steroidal saponin of F ring open loop.ESI-MS shows: 1053[M+Na]
+, point out this compound molecular weight should be: 1030.Molecular formula is C
51H
82O
21 1H-NMR and
13C-NMR sees Table 10 and table 11.With document [Liang Z Z etal, Planta Med.1988; 54 (4): 344] unanimity.
Table 10 compound (6)
1H-NMR (C
5D
5N, 400MHz)
| NO. | δ H/ppm | NO. | δ H/ppm |
| 17-H CH 3-18 CH 3-19 CH 3-21 CH 3-27 | 2.45(1H，d，J＝10.0) 0.72(3H，s) 1.04(3H，s) 1.61(3H，s，) 1.02(3H，d，J＝6.0) | 26-Glu 1 6 3-Glu 1 6 2’-Rha 1 6 4’-Rha 1 6 | 4.80(1H，d，J＝7.0) 4.34(1H，brd，J＝12.5) 4.42(1H，brd，J＝12.5) 4.95(1H，d，J＝7.0) 4.05(1H，brd，J＝10.4) 4.10(1H，brd，J＝10.4) 5.85(1H，brs) 1.64(3H，d，J＝6.0) 6.4(1H，brs) 1.75(3H，d，J＝6.0) |
Table 11 compound (6)
13C-NMR (C
5D
5N, 400MHz)
| NO. | δ C/ppm | NO. | δ C/ppm |
| 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 | 37.2 29.8 77.67 38.6 140.4 121.4 32.04 31.02 49.95 39.92 20.89 39.27 43.04 54.58 31.02 84.11 64.17 13.74 19.04 103.2 11.4 153.2 43.14 23.29 33.37 74.57 16.8 | 26-Glu 1 2 3 4 5 6 3-Glu 1 2 3 4 5 6 2’-Rha 1 2 3 4 5 6 4’-Rha 1 2 3 4 5 6 | 104.82 77.37 74.85 72.18 78.15 62.49 99.89 78.27 76.56 77.59 78.24 60.92 102.54 71.36 70.04 72.36 73.55 18.11 101.63 72.15 69.13 72.45 73.77 18.27 |
In the above-claimed cpd, compound (2) and (6) are known compound; Compound (1), (3), (4) and (5) are new compound.
Asparagus fern total steroid saponin preparation method of extract of the present invention has two kinds of following n-butanol extraction and macroreticular resin absorbing methods.Concrete preparation process is:
One, n-butanol extraction:
The asparagus fern meal is doubly measured the 70-80% ethanol percolate extraction with 20-30, gets medicinal extract.Medicinal extract dilutes with suitable quantity of water, and behind chloroform extraction, the aqueous solution is with n-butanol extraction, evaporate to dryness, and the residue dissolve with methanol adds acetone to precipitation and no longer produces, and filters, dry asparagus fern total steroid saponin extract.
The asparagus fern meal, with 20 times of amount 70～80% ethanol percolate extraction, decompression recycling ethanol gets medicinal extract.Medicinal extract is diluted with suitable quantity of water,, remove fat-soluble component with chloroform extraction.The aqueous solution is with n-butanol extraction 3-5 time, and the reclaim under reduced pressure propyl carbinol is to doing, and residue dissolves with small amount of methanol, add acetone under stirring and no longer produce to precipitation, filtration under diminished pressure, vacuum-drying gets asparagus fern total steroid saponin extract, yield is about 4-5%, and wherein content of furostanol saponin is about 70-80%.
Two, macroreticular resin absorbing method:
Get the asparagus fern meal, doubly measure 70～80% alcohol reflux 3 times with 7-15, the extracting solution reduction vaporization reclaims ethanol to there not being the alcohol flavor, behind the thin up with macroporous resin column (D101 macroporous resin column for example, the ZTC-1 macroporous resin column, AB-8 macroporous resin column etc.) carry out chromatographic separation, successively with water, 20% ethanol, the 70%-80% ethanol elution, to get asparagus fern total steroid saponin extract behind the 70%-80% ethanol elution part decompression recycling ethanol evaporate to dryness, yield is about 4～5%, and wherein content of furostanol saponin is 80～85%.
The asparagus fern total steroid saponin extract that the present invention makes with the piece root of asparagus fern through pharmacology activity research, finds that asparagus fern total steroid saponin extract has remarkable cardiovascular and cerebrovascular active function.The total steroid saponin extract is given anesthetized dog intravenous drip (5mg/Kg) test, finds that this total steroid saponin extract can significantly increase the anesthetized dog cerebral blood flow (CBF); Can significantly reduce the anesthetized dog cerebral vascular resistance; Anesthetized dog mean arterial pressure and heart rate there is not obvious influence; Quiet of dog (2.5mg/Kg) is given in the new compound of getting (1), (3), and specific activity total steroid saponin extract is better.Therefore, asparagus fern total steroid saponin extract can be used for preparation control cardiovascular and cerebrovascular medicine or food.
Embodiment
Below by embodiment, the invention will be further described.
Embodiment 1 n-butanol extraction:
Asparagus fern meal 1kg, with 20 times of amount 70～80% ethanol percolate extraction, the percolate decompression recycling ethanol gets medicinal extract.Medicinal extract is diluted with suitable quantity of water, with chloroform extraction 2 times (using chloroform 500ml) at every turn.The aqueous solution that chloroform extraction is crossed uses n-butanol extraction 4-5 time (to use propyl carbinol 400～500ml) more at every turn, merge butanol extraction liquid, the reclaim under reduced pressure propyl carbinol is to doing, residue dissolves with small amount of methanol, add acetone under stirring and no longer produce to precipitation, filtration under diminished pressure precipitates, and precipitation gets the total steroid saponin extract through vacuum-drying, yield is about 4%, and wherein content of furostanol saponin is about 75%.
Embodiment 2 macroreticular resin absorbing methods:
Asparagus fern meal 1kg, with 70～80% alcohol reflux 3 times (using ethanol 1000ml) at every turn, the alcohol extract reclaim under reduced pressure is not distinguished the flavor of to there being alcohol, behind the thin up, last D101 macroporous resin column is carried out chromatographic separation, successively with water, 20% ethanol, 70-80% ethanol elution, 70%-80% ethanol elution part decompression and solvent recovery is extremely done, residue promptly gets the total steroid saponin extract through vacuum-drying, and yield is about 5%, and wherein content of furostanol saponin is about 83%.
The cerebrovascular activity test of embodiment 3 asparagus fern total steroid saponin extracts
3.1 test objective: observation asparagus fern total steroid saponin extract, compound (1), compound (3) influence the anesthetized dog cerebrovascular.
3.2 be subjected to the reagent thing: see Table 12.
3.3 animal: 27 of healthy hybrid dogs are provided conformity certification number by the The 2nd Army Medical College Experimental Animal Center: SCXK (Shanghai) 2003-0002, body weight 13～15kg, the male and female dual-purpose, be divided at random 5 groups as follows, see Table 12:
Table 12
| Grouping | Number of animals (only) | Dosage | Route of administration |
| Negative control group | 6 | Physiological saline 5ml/kg | Intravenous drip |
| Nimodipine group asparagus fern total steroid saponin group compound (1) group compound (3) group | 6 6 5 4 | Nimodipine 300ug/kg 5mg/kg 2.5mg/kg 2.5mg/kg | Intravenous drip intravenous drip intravenous drip intravenous drip |
3.4 test method: animal is used Nembutal vein anesthetic (30mgkg
-1); Separate right common carotid artery and external carotid artery, after ligation external carotid artery and other arteriole branches, laying diameter on arteria carotis communis is the magnetic flow meter probe of 3mm, with MFV-3200 type Electromagnetic Flow instrument (Japanese photoelectricity Co., Ltd.) record volume of blood flow, and expression ICAF amount.Separate femoral artery, parallel intubate is thrust four limbs and the front is subcutaneous with needle electrode, and the FAP force transducer is connected bioelectrical signals amplifier (productions of Shanghai Alcott company) and by the computer demonstration, writes down arteriotony with cardiac diagnosis lead-line; Separate femoral vein, parallel intubate (intravenously administrable is used).Separate tracheae, insert trachea cannula.After operation finishes, stablized 30 minutes, write down once every index as administration before control value; The administration volume is 5ml/kg, and the quiet notes time is 30min.Quiet notes back was write down every desired value in 5,15,30,60,90,120 minutes respectively.Put to death dog after 120 minutes, get brain and weigh.Multiply by 2 representative full cerebral blood flow (CBF)s (CBF) with the right common carotid artery volume of blood flow.Cerebral vascular resistance can be calculated with formula: R=MBP (kPa)/[CBF (ml/min) 100g brain].
3.5 route of administration: intravenous drip.
3.6 administration number of times: equal single-dose.
3.7 observation index and observing time: observed and recorded ICAF amount, blood pressure, be after the administration 5,15,30,60,90,120 minutes observing times.
3.8 data and statistical procedures:
Experimental result with
Expression adopts one-tenth team two sample analyses of t check paired value to do administration front and back statistical study, comparative analysis between variance t such as two samples check work group, and P＜0.05 is judged as the significant difference with significance.
3.9 observed content:
1, to the influence of cerebral blood flow (CBF): to the influence of cerebral blood flow (CBF), its velocity of variation and physiological saline control group relatively have there was no significant difference with time point before and after the medication.
2, to the influence of cerebral vascular resistance: the variation of cerebral vascular resistance before and after the medication, its velocity of variation and physiological saline control group relatively have there was no significant difference with time point.
3, to the influence of mean blood pressure: to the influence of mean blood pressure, velocity of variation and physiological saline control group before and after the administration have not statistically significant with the time point comparing difference before and after the medication.
4, to the influence of heart rate: to the influence of heart rate, its velocity of variation and physiological saline control group relatively have there was no significant difference with time point before and after the medication.
3.10 experimental result:
1, the intravenous drip of asparagus fern total steroid saponin extract is to the influence of anesthetized dog cerebral blood flow (CBF): before the medication of physiological saline control group with medication after each time point comparison cerebral blood flow (CBF) slightly reduce and cerebral blood flow (CBF) in 30 minutes, raise (P＜0.05) relatively before and after the asparagus fern total steroid saponin extract, compound (1), the medication of compound (3) group.And velocity of variation between physiological saline group comparative group, asparagus fern total steroid saponin extract, compound (1), compound (3) group raise in 30 minutes obviously, (P＜0.05-0.01).See Table 13.
2, to the influence of anesthetized dog cerebral vascular resistance
Before the medication of physiological saline group relatively cerebral vascular resistance do not have considerable change, after asparagus fern total steroid saponin extract, compound (1), the medication of compound (3) group with medication before relatively cerebral vascular resistance in 60min, reduce.Its velocity of variation and physiological saline group relatively obviously reduced (P＜0.05～0.01) and see Table 14 in 30 minutes.
3, to the influence of anesthetized dog mean blood pressure and heart rate
Physiological saline group and asparagus fern total steroid saponin extract, compound (1), compound (3) group all do not have obvious influence to anaesthetized dog blood pressure and heart rate.See Table 15 and 16.
The influence of table 13 pair anesthetized dog cerebral blood flow (CBF) (ml/min.100g,
N=6)
| Group | Before the administration | After the administration | |||||
| 5min | 15min | 30min | 60min | 90min | 120min | ||
| The physiological saline group | 428.45.52±122.81 | 405.07 ±140.47 | 409.08±126.98 | 399.31±119.82 | 387.87±70.28 | 321.49±54.86 | 290.010±80.64 |
| Rate of change % Nimodipine group rate of change % Radix asparagi steroid saponin extract group rate of change % compound (1) group rate of change % compound (3) group rate of change % | 299.46±35.85 265.15±79.96 371.75±30.96 381.54±37.43 | -5.97±16.14 334.82±61.90 13.11±24.56 338.10±98.70 △△ 28.32±16.29 ** 445.96±70.72 21.44±26.67 450.03±40.45 18.14±642 | -4.89±9.95 339.06±44.57 14.37±18.84 * 310.40±103.29 △ 16.70±12.67 * 419.41±58.72 14.12±23.59 427.36±49.06 12.06±8.03 | -20.10±35.43 331.95±20 42 12.04±13.87 ** 280.21±101.48 4.58±13.27 * 357.89±35.38 -2.74±16.16 404.01±38.84 6.15±8.77 | -6.51±17.06 355.02±29.94 * 19.96±17.22 * 227.66±114.58 -17.30±23.00 329.25±44.23 -10.39±17.85 327.17±30.63 -13.20±15.09 | -22.30±14.05 332.65±60.56 10.98±14.23 ** 206.69±122.78 -26.41±48 280.63±41.13 -24.08±13.15 285.15±33.37 -24.10±15.95 | -30.00±19.18 322.23±69.21 6.91±14.60 ** 195.48±113.01 -30.00±24.83 217.55±43.17 -41.77±8.00 224.26±60.59 -39.65±21.72 |
Velocity of variation and physiological saline control group compare with time point,
*P＜0.05,
*P＜0.01;
△P＜0.05,
△ △Compare before and after group innerlich anwenden of p＜0.01
The influence of table 14 pair anesthetized dog cerebral vascular resistance (the kPamin/ml100g brain,
N=6)
| Group | Before the administration | After the administration | |||||
| 5min | 15min | 30min | 60min | 90min | 120min | ||
| The physiological saline group | 4.01±1.67 | 4.22±1.36 | 4.17±1.35 | 4.17±1.15 | 4.27±0.73 | 5.72±1.35 | 6.92±1.94 |
| Velocity of variation % | 7.00±18.07 | 7.59±20.80 | 8.52±19.94 | 18.96±35.10 | 55.05±47.32 | 85.79±66.48 | |
| The nimodipine group | 5.09±1.92 | 2.60±0.77 △ | 2.65±0.66 △ | 3.11±0.65 △ | 3.36±0.92 △ | 4.32±2.08 | 4.45±2.25 |
| Rate of change % Radix asparagi steroid saponin extract group rate of change % compound (1) group rate of change % compound (3) group rate of change % | 5.26±1.13 7.11±0.91 6.77±1.57 | -57.98±10.92 * 4.12±0.76 -20.67±12.49 * 5.87±0.92 -17.19±9.89 * 5.66±1.49 -16.78±3.71 * | -38.62±36.63 * 4.37±1.08 -16.42±17.51 * 5.92±0.65 -16.04±11.57 * 5.77±1.07 -14.02±4.79 * | -28.66±39.56 * 4.40±1.74 -17.38±23.84 * 7.40±0.69 4.74±9.89 6.14±2.15 -10.99±11.82 * | -22.65±46.17 * 5.25±2.44 4.80±37.35 7.42±0.56 5.31±12.15 7.76±2.25 14.33±25.67 | -10.31±33.84 * 7.17±3.86 36.78±75.32 8.23±0.90 17.58±20.77 9.02±2.50 33.12±26.52 | -9.18±33.21 * 8.62±4.63 62.45±87.00 11.77±2.94 66.03±40.80 11.29±3.87 66.74±51.48 |
Velocity of variation and solvent control group compare with time point,
*P＜0.05,
*P＜0.01
The influence of table 15 pair anesthetized dog mean blood pressure (kPa,
, n=6)
| Group | Before the administration | Time after the administration | |||||
| 5min | 15min | 30min | 60min | 90min | 120min | ||
| The physiological saline group | 15.43±3.76 | 14.78±2.07 | 15.15±1.83 | 15.12±1.31 | 15.93±1.58 | 17.50±2.95 | 18.18±2.53 |
| Velocity of variation % | -0.68±21.12 | 2.53±24.23 | 2.65±23.53 | 7.73±24.22 | 17.79±31.02 | 21.88±26.46 | |
| The nimodipine group | 14.85±4.69 | 8.63±2.59 △ | 8.87±2.05 △ | 10.28±1.98 | 12.12±2.27 | 13.53±3.75 | 13.22±3.61 |
| The total soap body saponin extract of rate of change % asparagus fern group rate of change % compound (1) group rate of change % compound (3) group rate of change % | 13.55±3.10 17.44±1.36 17.40±4.44 | -34.15±41.15 13.57±3.24 0.49±10.34 18.12±1.52 0.04±0.05 17.00±4.14 -1.78±5.42 | -33.05±34.37 12.95±3.18 -3.80±13.55 16.96±1.28 -0.02±0.10 16.50±3.43 -3.52±11.16 | -22.09±37.22 11.40±3.26 -16.20±12.02 18.56±1.36 0.07±0.11 16.30±4.37 -5.92±10.05 | -8.96±39.59 10.18±3.19 -23.64±19.14 17.08±1.44 -0.02±0.07 16.85±4.59 -3.64±3.58 | -2.69±29.46 11.40±2.99 -14.38±19.41 16.18±2.25 -0.07±0.15 16.98±4.14 -2.06±3.11 | -4.28±31.73 12.73±3.70 -3.75±28.92 17.34±1.23 0.00±0.12 15.90±3.44 -7.49±7.35 |
Compare before and after △ P＜0.05, medication.
The influence of table 16 pair anesthetized dog heart rate (inferior/minute)
| Group | Before the administration | After the administration | |||||
| 5min | 15min | 30min | 60min | 90min | 120min | ||
| The physiological saline group | 202±37 | 178±26 | 184±19 | 188±18 | 184±30 | 179±53 | 203±35 |
| Velocity of variation % | -9±23 | -6±20 | -5±17 | -7±16 | -11±25 | 2±15 | |
| The nimodipine group | 182±32 | 157±38 | 166±34 | 155±45 | 166±40 | 172±38 | 180±34 |
| Rate of change % Radix asparagi steroid saponin extract group rate of change % compound (1) group rate of change % compound (3) group rate of change % | 173±36 205±36 199±27 | -12.96±19.31 163±31 -5±5 190±18 -0.06±0.10 179±36 -11±11 | -7.94±18.72 158±29 -8±10 186±13 -0.08±0.12 185±48 -8±16 | 13.78±25.46 153±23 -10±13 184±13 -0.09±0.12 168±37 -16±10 | -7.21±21.94 148±23 -13±10 170±21 -0.16±0.12 170±55 -16±17 | -3.35±24.78 152±28 -11±10 175±30 -0.14±0.17 166±43 -18±11 | 1.25±23.31 146±38 -15±12 174±34 -0.14±0.21 153±46 -24±15 |
Conclusion
The test-results of above embodiment 3 shows:
1. asparagus fern total steroid saponin extract, compound (1) and compound (3) intravenous drip can obviously increase the anesthetized dog cerebral blood flow (CBF).
2. asparagus fern total steroid saponin extract, compound (1) and compound (3) intravenous drip have obvious reduction anesthetized dog cerebral vascular resistance effect.
3. asparagus fern total steroid saponin extract, compound (1) and compound (3) intravenous injection do not have obvious influence to anesthetized dog mean arterial pressure and heart rate.
Test-results shows that asparagus fern total steroid saponin extract, compound (1) and compound (3) have as the primary condition of effectively preventing and treating the medicine of cardiovascular and cerebrovascular diseases.Therefore, total steroid saponin extract, compound (1) and the compound (3) of Chinese medicine asparagus fern preparation of the present invention can be used for preparation control cardiovascular and cerebrovascular medicine or food.
Claims (4)
1, a kind of asparagus fern total steroid saponin extract is characterized in that content of furostanol saponin is 70-85% in the described extract, and described extract contains 6 kinds of compounds, and wherein 4 kinds of compounds are:
Compound (1): (25S)-26-O-β-D-glucopyanosyl base-5 β-furan steroid-20 (22)-alkene-3 β, 26-glycol-3-O-α-L-sandlwood pyrans glycosyl (1 → 4)]-β-D-glucopyranoside;
Compound (3): (25R)-26-O-β-D-glucopyanosyl base-furan steroid-5,20-diene-3 β, 26 glycol-3-O-[α-L-sandlwood pyrans glycosyl (1 → 2)]-[(β-D-glucopyanosyl-(1 → 4))-α-L-sandlwood pyrans glycosyl-(1 → 4)]-β-D-glucopyranoside;
Compound (4): (25S)-26-O-β-D-glucopyanosyl base-5 β-furan steroid-20 (22)-alkene-3 β, 15,26-triol-3-O-α-L-sandlwood pyrans glycosyl (1 → 4)]-β-D-glucopyranoside;
Compound (5): (25S)-26-O-β-D-glucopyanosyl base-5 β-furan steroid-20 (22)-alkene-3 β, 26-glycol-3-O-[β-D-glucopyanosyl base-(1 → 2)]-β-D-glucopyranoside.
2, the described asparagus fern total steroid saponin of claim 1 preparation method of extract is characterized in that preparation process is: the asparagus fern meal is doubly measured the 70-80% ethanol percolate extraction with 20-30, gets medicinal extract with the n-butanol extraction preparation; Medicinal extract dilutes with suitable quantity of water, and behind chloroform extraction, the aqueous solution is with n-butanol extraction, evaporate to dryness, and the residue dissolve with methanol adds acetone to precipitation and no longer produces, and filters, dry asparagus fern total steroid saponin extract.
3, the described asparagus fern total steroid saponin of claim 1 preparation method of extract, it is characterized in that preparing with macroreticular resin absorbing method, preparation process is: get the asparagus fern meal, 70-80% alcohol reflux so that 7-15 doubly measures boils off ethanol, carries out chromatographic separation with macroporous resin column, successively with water, 20% ethanol, the 70%-80% ethanol elution gets asparagus fern total steroid saponin extract with 70-80% ethanol elution part evaporate to dryness.
4, the application of the described asparagus fern total steroid saponin of claim 1 extract is characterized in that described asparagus fern steroidal saponin extract is in preparation control cardiovascular and cerebrovascular medicine or Application in Food.
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| CN101307090B (en) * | 2007-05-16 | 2011-04-13 | 中国科学院上海药物研究所 | Method for preparing timosaponin BIII and uses thereof |
| EP2194998A4 (en) * | 2007-09-07 | 2012-03-21 | Bionovo Inc | Estrogenic extracts of asparagus conchinchinensis (lour.) merr of the liliaceae family and uses thereof |
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0348694A (en) * | 1989-07-18 | 1991-03-01 | Dowa Mining Co Ltd | Novel saponin substance |
| CN1102186A (en) * | 1993-10-29 | 1995-05-03 | 沈阳医学院 | Determination and usage of new component of traditional Chinese medicine Chinese onion |
| CN1212966A (en) * | 1997-09-26 | 1999-04-07 | 中国人民解放军军事医学科学院放射医学研究所 | Usage of steroi saponin for preventing and curing senile dementia and new steroid saponin |
| WO2003080067A1 (en) * | 2002-03-21 | 2003-10-02 | Council Of Scientific And Industrial Research | 3-0-'alfa-l-rhamnopyranosyl- (1-2) -alpha-l-rhamnopyranosyl- (1-4) -3beta-d-glucopyranosyl!-25 (s) -spirost an-3beta-ol, process for its isolation, and its use as immunomodulator |
| KR20030091319A (en) * | 2002-05-27 | 2003-12-03 | 대한민국 (관리부서 농촌진흥청) | New anticancer compounds and its purification from Asparagus oligoclonos |
| US6670459B2 (en) * | 2002-03-20 | 2003-12-30 | Zandu Pharmaceutical Works Ltd. | Process for the isolation of novel oligospirostanoside |
-
2004
- 2004-05-09 CN CNB2004100181802A patent/CN1315866C/en not_active Expired - Fee Related
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0348694A (en) * | 1989-07-18 | 1991-03-01 | Dowa Mining Co Ltd | Novel saponin substance |
| CN1102186A (en) * | 1993-10-29 | 1995-05-03 | 沈阳医学院 | Determination and usage of new component of traditional Chinese medicine Chinese onion |
| CN1212966A (en) * | 1997-09-26 | 1999-04-07 | 中国人民解放军军事医学科学院放射医学研究所 | Usage of steroi saponin for preventing and curing senile dementia and new steroid saponin |
| US6670459B2 (en) * | 2002-03-20 | 2003-12-30 | Zandu Pharmaceutical Works Ltd. | Process for the isolation of novel oligospirostanoside |
| WO2003080067A1 (en) * | 2002-03-21 | 2003-10-02 | Council Of Scientific And Industrial Research | 3-0-'alfa-l-rhamnopyranosyl- (1-2) -alpha-l-rhamnopyranosyl- (1-4) -3beta-d-glucopyranosyl!-25 (s) -spirost an-3beta-ol, process for its isolation, and its use as immunomodulator |
| KR20030091319A (en) * | 2002-05-27 | 2003-12-03 | 대한민국 (관리부서 농촌진흥청) | New anticancer compounds and its purification from Asparagus oligoclonos |
Non-Patent Citations (1)
| Title |
|---|
| 呋喃甾烷型皂甙的研究进展 张剑波等，有机化学，第20卷第5期 2000 * |
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