CN101239093A

CN101239093A - Active component in albizia julibrissin with inhibiting angiogenesis function and preparation method and application thereof

Info

Publication number: CN101239093A
Application number: CNA2008100204963A
Authority: CN
Inventors: 金坚; 花慧; 张莲芬; 冯磊; 陆核; 李红
Original assignee: Jiangnan University
Current assignee: Jiangnan University
Priority date: 2008-03-10
Filing date: 2008-03-10
Publication date: 2008-08-13

Abstract

The invention relates to an active ingredient which has role in inhibiting angiogenesis in cortex albiziae. The active ingredient is a saponin compound which has applications of treatment and prevention of diseases about angiogenesis and anti-tumor angiogenesis, can be used in medicines which can prevents angiogenesis, or can be used as conventional medicine carrier and/or excipient, or is applied in preparation of health-care food or cosmetics for prevention of angiogenesis, so that the invention also provides the preparation method of the active ingredient, which is that the dry tree barks, stem skins, leaves, flowers, seeds, pods, or roots of the cortex albiziae are extracted by organic solvents, organic solvents with water or water; the extracting solution is filtered and condensed into extract; the extract is eluted and separated by a D-101 shape macroporous resin, then the effective groups are collected and are eluted and separated by a silica gel chromatography column, another effective groups are collected and separated by an RP-styrene column so that an effective group is obtained, the solvent is recycled to be an extract which is freezed, dried and then made into drymeal.

Description

Has active ingredient that suppresses angiogenesis function and its production and application in the Cortex Albiziae

(1) technical field

The present invention relates to have the active ingredient that suppresses angiogenesis function in the Cortex Albiziae, the invention still further relates to the preparation method and the application of described active ingredient.

(2) background technology

Angiogenesis (angiogenesis) refers to tissue utilization and had both deposited the process that blood vessel produces new blood vessel.The vascular endothelial cell of normal adult generally is in relative static conditions, the average per several years division of endotheliocyte once under the physiological conditions, only betide the ovary and the endometrium of repair in trauma or menstrual cycle, but numerous disease such as tumor, the quick hypertrophy of new vessels endotheliocyte in can occurring organizing, its hypertrophy cycle is equivalent to the germinal cell or the intestinal epithelial cell [1 of bone marrow, T í m á r J, D  me B, Fazekas K, Janovics A, PakuS.Angiogenesis-dependent diseases and angiogenesis therapy.PatholOncol Res.2001,7 (2): 85-94.2, Folkman J.Angiogenesis in cancer, rheumatoid and disease.Nat Med.1995,1:27-31.], the growth of most malignant entity tumors and transfer are that blood vessel relies on, be that tumor tissues can be induced new angiogenesis in its growth course, so that required nutrition of tumor growth and oxygen to be provided, carry transitional cell by blood vessel to its hetero-organization simultaneously, and form in other position continued growths of body and induction of vascular, cause neoplasm metastasis [3, D ' Amato RJ, Adamis AP.Angiogenesis inhibition in age-related macular degeneration.Ophthalmology. 1995,102 (9): 1261-1262.4, Arbiser JL.Angiogenesis andthe skin:a primer.J Am Acad Dermatol.1996,34 (3): 486-497.5, O ' BrienKD, McDonald TO, Chait A, Allen MD, Alpers CE.Neovascular expressionof E-selectin, intercellular adhesion molecule-1, and vascular celladhesion molecule-1 in human atherosclerosis and their relation tointimal Leukocyte content.Circulation.1996,93 (4): 672-82.6, HanahanD, Folkman J.Patterns and emerging mechanisms of the angiogenic switchduring tumorigenesis.Cell.1996,86 (3): 353-364.].Think at present, cancerous cell can be secreted a large amount of many angiogenesis factors that promote angiogenic growth, make tumor form new blood vessel and growth fast, and vascular endothelial cell also can be secreted angiogenesis factor and impels the propagation of endothelial cell self and even tumor cell [7, Folkman, Cotran.Relation of vascular proliferation to tumor growth.Int Rev Exp Pathol.1976,16:207-248.8, Veikkola T, Alitalo K.VEGFs, receptors and angiogenesis.Semin Cancer Biol.1999,90:211-220.] generally speaking, the generation and the transfer of angiogenesis and tumor have confidential relation.The generation of inhibition new vessels has become a neofield in the treatment and prevention of tumour.U.S. Folkman seminar in 1998 reports that angiostatin (Angiostatin) and Endostatin (Endostatin) share, mice to the big intestinal tumour of lotus, prostate tumor, mastadenoma and cerebroma has significant curative effect, and Endostatin is not developed immunity to drugs, thereby cause the extensive concern of scientific circles, make angiogenesis inhibitor treatment tumor become a big focus.

To suppress angiogenesis is target spot, thereby reaches the purpose of the transfer that suppresses tumor, and this neodoxy has overcome the drawback of treatment tumor in the past, is difficult for producing drug resistance.The effect of medicine can be to normal cell toxigenicity.Countries in the world are passed through broad research with tumor-blood-vessel growth as target spot, develop many angiogenesis inhibitors at present and carrying out the I～research of III clinical trial phase, for example kinds more than 20 such as angiogenesis chalone (Angiostatin), Endostatin (Endostatin), Amebacilin analog and metalloprotein enzyme inhibition factor and urokinase, il-1 2, platelet factor, Bufotanine.Above-mentioned angiogenesis inhibitor has high specificity, and effect is fast, and dosage is little, the curative effect height, and side effect is little, is difficult for producing advantages such as drug resistance.This type of medicine not only can be used for the treatment of most of entity tumors, also can be used for the prevention of cancer and the treatment of blood system malignant tumor, simultaneously to the disease of other and associated angiogenesis as: the prevention and the treatment of diabetic renal papillary necrosis, rheumatic arthritis, psoriasis, hemangioma, atherosclerosis etc. all have important theory and realistic meaning.

It is pulse family Mimosoideae plant that Cortex Albiziae belongs to albizzia Albizzia, is distributed widely in all over the world.China has 16 kinds, major part originates in the south and the west and south, wherein modal have a Herba Albiziae AlbizziaJulibrissin Durazz, principal columns of a hall tree Albizzia chinensis (Osb.) Merr., lebbek Albizzia Lebbeck (L.) Benth and BAIGE Aibizzia Procera (L.) Benth etc.The main chemical constituent of albizzia plant bark, peel of stem, leaf, flower, seed has triterpene and glycosides compound, flavone and glycosides compound thereof, alkaloid, organic acid, phytosterin compound, lignanoid, tannin and volatile ingredient etc.Modern pharmacology experiment shows, effects such as that albizzia plant has is anticancer widely, calm, antifertility, paf receptor antagonism and antiinflammatory.For example, by the Cortex Albiziae in albizzia plant of including of Chinese Pharmacopoeia is conventional Chinese medicine simply, dry bark for leguminous plant Herba Albiziae Albizia julibrissin Durazz., has resolving stagnation for tranquilization, the function of promoting blood circulation and detumescence is used for irritability, melancholy insomnia, the lung abscess skin ulcer is swollen, falls diseases such as pouncing on the pain of injury.Cortex Albiziae extract has widely in the body, the activity of extracorporeal suppression tumor cell propagation, but because its antitumaous effect mechanism is also indeterminate, limited the clinical practice [9 of its antitumaous effect, TsuyoshiIkeda, Tetsuya Kajimoto, Toshihiro Nohara, Jun-ei Kinjo, Chi-Huey Wong.Preparation of a neoglycolipid carrying the oligosaccharide componentof saponin from Albizzia julibrissin.Tetrahedron Letters.1995,36 (9): 1509-1510.10, Kun Zou, Yuying Zhao, Guangzhong Tu, JingrongCui, Zhonghua Jia and Ruyi Zhang.Two diastereomeric saponins withcytotoxic activity from Albizia julibrissin.Carbohydrate Research.2000,324 (3): 182-188.11, Kun Zou, Yuying Zhao, Guangzhong Tu, JunhuaZheng, Ruyi Zhang.A New Isomer of Julibroside J ₂From Albizia julibrissin.J Asian Natural Products Research.1998,1 (1): 59-66.12, Zou Kun, Zhao Yuying, Li Deyu, Xu Feng, Zheng Junhua, Zhang Ruyi. Hydrophobic constituents from Albizia julibrissin. journal of Beijing Medical University, 1999,31 (1): 32-34.] albizzia Albizzia is a pulse family Mimosoideae plant, is distributed widely in all over the world.China has 16 kinds, major part originates in the south and the west and south, wherein modal have a Herba Albiziae Albizzia Julibrissin Durazz, principal columns of a hall tree Albizzia chinensis (Osb.) Merr., lebbek Albizzia Lebbeck (L.) Benth and BAIGE Albizzia Procera (L.) Benth etc.The main chemical constituent of albizzia plant bark, peel of stem, leaf, flower, seed has triterpene and glycosides compound, flavone and glycosides compound thereof, alkaloid, organic acid, phytosterin compound, lignanoid, tannin and volatile ingredient etc.Modern pharmacology experiment shows, effects such as that albizzia plant has is anticancer widely, calm, antifertility, paf receptor antagonism and antiinflammatory.For example, by the Cortex Albiziae in albizzia plant of including of Chinese Pharmacopoeia is conventional Chinese medicine simply, dry bark for leguminous plant Herba Albiziae Albizia julibrissin Durazz., has resolving stagnation for tranquilization, the function of promoting blood circulation and detumescence is used for irritability, melancholy insomnia, the lung abscess skin ulcer is swollen, falls diseases such as pouncing on the pain of injury.Cortex Albiziae extract has widely in the body, the activity of extracorporeal suppression tumor cell propagation, but because its antitumaous effect mechanism is also indeterminate, limited the clinical practice [13 of its antitumaous effect, TsuyoshiIkeda, Tetsuya Kajimoto, Toshihiro Nohara, Jun-ei Kinjo, Chi-Huey Wong.Preparation of a neoglycolipid carrying the oligosaccharide componentof saponin from Albizzia julibrissin.Tetrahedron Letters.1995,36 (9): 1509-1510.14, Kun Zou, Yuying Zhao, Guangzhong Tu, JingrongCui, Zhonghua Jia and Ruyi Zhang.Two diastereomeric saponins withcytotoxic activity from Albizia julibrissin.Carbohydrate Research.2000,324 (3): 182-188.15, Kun Zou, Yuying Zhao, Guangzhong Tu, JunhuaZheng, Ruyi Zhang.A New Isomer of Julibroside J ₂From Albizia julibrissin.J Asian Natural Products Research.1998,1 (1): 59-66.16, Zou Kun, Zhao Yuying, Li Deyu, Xu Feng, Zheng Junhua, Zhang Ruyi. Hydrophobic constituents from Albizia julibrissin. journal of Beijing Medical University, 1999,31 (1): 32-34.]

Multicomponent, many target spots, multipath " be the characteristics of treatment by Chinese herbs disease, performance motherland medical science characteristic, from multipath, multi-level, too many levels, research Chinese medicine directly or affect indirectly angiogenesis has important clinical meaning and application prospect.

(3) summary of the invention

At the problems referred to above, the invention provides have in the Cortex Albiziae suppress angiogenesis function active ingredient its can be used to suppress tumor vessel and other and angiogenesis equally, the present invention also provides the preparation method and the application of described active ingredient.

Technical scheme of the present invention:

Having the active ingredient that suppresses angiogenesis function in the Cortex Albiziae is oside compound.

Described oside compound wherein contains compd A and/or compd B, being characterized as of the molecular structure of described compd A:

(I) R1:CH wherein ₃R ₂: NHAC R ₃: OH C-6:R

The molecular formula of compd A is: C ₁₀₄H ₁₆₅O ₄₉N, molecular weight is 2211, fusing point is 202～204 ℃, chemistry 3-o-[β by name-D-xylopyranosyl-(1 → 2)-β-D-pyrans furan glycosyl-(1 → 6)-β-D-2-deoxidation-2-acetamido glucosyl group]-21-o-{ (6s)-2-is trans-and ((6R)-2-trans-2 for 2-methylol-6-methyl-6-o-[4-o-, 6-dimethyl-6-o-β-D-pyrans quinovose base-2,7-octadiene acidic group)-and β-D-pyrans quinovose base]-2,7-octadiene acidic group }-acacic acid-28-o-β-D-glucopyranosyl-(1 → 3)-[α-L-arabinofuranosyl-(1 → 4)]-α-L-rhamnopyranosyl-(1 → 2)-β-D-glucopyranosyl ester;

Being characterized as of the molecular structure of described compd B:

(II) R wherein ₁: CH ₃R ₂: OH C-6:S

The molecular formula of compd B is: C ₁₀₂H ₁₆₂O ₄₉, molecular weight is 2170, fusing point is 204～206 ℃; Chemical name 3-o-[β-D-xylopyranosyl-(1 → 2)-β-D-pyrans furan glycosyl-(1 → 6)-β-D-glucopyranosyl]-21-o-{ (6s)-2-is trans-and ((6R)-2-trans-2 for 2-methylol-6-methyl-6-o-[4-o-, 6-dimethyl-6-o-(β-D-pyrans quinovose base)-2,7-octadiene acidic group)-and β-D-pyrans quinovose base]-2,7-octadiene acidic group }-acacic acid-28-o-β-D-pyrans Fructus Vitis viniferae-(1 → 3)-[α-L-arabinofuranosyl-(1 → 4)]-α-L-rhamnopyranosyl-(1 → 2)-β-D-glucopyranosyl ester.

The preparation method that has the active ingredient that suppresses angiogenesis function in the Cortex Albiziae, it is characterized in that: with dry bark, peel of stem, leaf, flower, seed, soybean pod or the root of organic solvent, water-containing organic solvent or water extraction Cortex Albiziae, and with extracting liquid filtering, be condensed into extractum; The extractum employing D-101 type macroporous resin eluting of gained separates, and collects active princlple and carries out the separation of silica gel column chromatography eluting, regathers active princlple and carries out anti-phase styrene chromatographic column separation, gets active princlple, reclaims solvent and becomes extractum, and dry powder is made in lyophilization.

It is further characterized in that: separate described anti-phase styrene chromatographic column to such an extent that active princlple carries out getting oside compound A and oside compound B after anti-phase C18 chromatographic column eluting separates.

Have the application of the active ingredient that suppresses angiogenesis function in the Cortex Albiziae, it is characterized in that: described oside compound has antineoplastic vascular to generate and the purposes of other and associated angiogenesis disease treatment, prevention; Or in the medicine that suppresses angiogenesis, use; Perhaps as conventional pharmaceutical carrier and/or excipient; The perhaps application in the health food of preparation inhibition angiogenesis; The perhaps application in the cosmetics of preparation inhibition angiogenesis.

Its further feature is: when making described inhibition angiogenesis drug, can add one or more pharmaceutically acceptable carriers; Described carrier comprises the diluent of pharmaceutical field routine, excipient, filler, binding agent, wetting agent, disintegrating agent, absorption enhancer, surfactant, absorption carrier, lubricant, emulsifying agent, osmotic pressure regulator can also add flavouring agent, sweeting agent etc. in case of necessity; When making described inhibition angiogenesis drug, preparation process that can be routinely, use separately or prepare the medicine of operable various different dosage forms clinically, for example various ways such as injection, tablet, powder, granule, capsule, oral liquid, unguentum, cream or Emulsion with other medicines; The described inhibition angiogenesis drug of making is used to prevent and/or treat at least a disease that is selected from one of the following: cancer and/or cancer metastasis, hemangioma, fibrohemangioma, neovascular glaucoma, diabetic renal papillary necrosis, retinopathy of prematurity and retinal vein occlusion, retinal degeneration, retrolental fibroplasia, trachomatous conjunctivitis, the keratopathy that angiogenesis causes, climacteric macula lutea and degeneration of macula, psoriasis, degeneration of old plate-like speckle and rheumatic arthritis, psoriasis, telangiectasis, botryomycosis hominis suppresses spot and forms in the wound healing, atherosclerosis, seborrheic dermatitis and acne; The described inhibition angiogenesis drug of making is used for the purposes of chemotherapy of tumors and/or adjuvant chemotherapy.

Beneficial effect of the present invention:

Having the active ingredient that suppresses angiogenesis function in the clear and definite first Cortex Albiziae of the present invention is oside compound, and with it as the disease prevention that is used for tumor-blood-vessel growth and other and associated angiogenesis, the medicine of treatment, from multicomponent, many target spots, multipath, at many levels, the too many levels aspect directly or affect indirectly angiogenesis, have important clinical meaning and application prospect, with the angiogenesis theory is the research background, for setting forth the mechanism of action of Chinese medicine from now on, the direction that developing Chinese medicine pharmacology opinion is new finds that new drug target provides important scientific basis.

(4) description of drawings

The anti-phase Resource column chromatography figure of Fig. 1: 298-J--D-G-R

The anti-phase C of Fig. 2: 298-J-D-G-R2-A, 298-J-D-G-R2-B (compd A, compd B) ₁₈Liquid chromatogram

Fig. 3: the separation and purification figure frame diagram of the active princlple of Cortex Albiziae, compd A, compd B

Fig. 4: the liquid chromatogram of compd A

Fig. 5: the liquid chromatogram of compd B

The mass spectrum of Fig. 6: 298-J-D-G

The mass spectrum of Fig. 7: 298-J--D-G-R

Fig. 8: compd B is to the inhibitory action figure of human vascular endothelial transfer ability

Fig. 9: compd B is to the inhibitory action figure of Matrigel tube method blood vessel in the body

Figure 10: compd B suppresses chick chorioallantoic membrane angiogenesis figure (CAM experiment)

Figure 11: the mass spectrum of compd A

Figure 12: the carbon spectrogram of compd A

Figure 13: the hydrogen spectrogram of compd A

Figure 14: the infrared spectrum of compd A

Figure 15: the mass spectrum of compd B

Figure 16: the carbon spectrogram of compd B

Figure 17: the hydrogen spectrogram of compd B

Figure 18: the infrared spectrum of compd B

(5) specific embodiment

(I) R wherein ₁: CH ₃R ₂: NHAC R ₃: OH C-6:R

The molecular formula of compd A is: C ₁₀₄H ₁₆₅O ₄₉N, molecular weight is 2211, fusing point is 202～204 ℃, chemistry is by name: 3-o-[β-D-xylopyranosyl-(1 → 2)-β-D-pyrans furan glycosyl-(1 → 6)-β-D-2-deoxidation-2-acetamido glucosyl group]-21-o-{ (6s)-2-is trans-and ((6R)-2-trans-2 for 2-methylol-6-methyl-6-o-[4-o-, 6-dimethyl-6-o-β-D-pyrans quinovose base-2,7-octadiene acidic group)-and β-D-pyrans quinovose base]-2,7-octadiene acidic group }-acacic acid-28-o-β-D-glucopyranosyl-(1 → 3)-[α-L-arabinofuranosyl-(1 → 4)]-α-L-rhamnopyranosyl-(1 → 2)-β-D-glucopyranosyl ester;

Being characterized as of the molecular structure of described compd B:

(II) R wherein ₁: CH ₃R ₂: OH C-6:S

The application that has the active ingredient that suppresses angiogenesis function in the Cortex Albiziae, described oside compound have antineoplastic vascular to generate and the purposes of other and associated angiogenesis disease treatment, prevention; Or in the medicine that suppresses angiogenesis, use; Perhaps as conventional pharmaceutical carrier and/or excipient; The perhaps application in the health food of preparation inhibition angiogenesis; The perhaps application in the cosmetics of preparation inhibition angiogenesis.

When making described inhibition angiogenesis drug, can add one or more pharmaceutically acceptable carriers; Described carrier comprises the diluent of pharmaceutical field routine, excipient, filler, binding agent, wetting agent, disintegrating agent, absorption enhancer, surfactant, absorption carrier, lubricant, emulsifying agent, osmotic pressure regulator can also add flavouring agent, sweeting agent etc. in case of necessity; When making described inhibition angiogenesis drug, preparation process that can be routinely, use separately or prepare the medicine of operable various different dosage forms clinically, for example various ways such as injection, tablet, powder, granule, capsule, oral liquid, unguentum, cream or Emulsion with other medicines; The described inhibition angiogenesis drug of making is used to prevent and/or treat at least a disease that is selected from one of the following: cancer and/or cancer metastasis, hemangioma, fibrohemangioma, neovascular glaucoma, diabetic renal papillary necrosis, retinopathy of prematurity and retinal vein occlusion, retinal degeneration, retrolental fibroplasia, trachomatous conjunctivitis, the keratopathy that angiogenesis causes, climacteric macula lutea and degeneration of macula, psoriasis, degeneration of old plate-like speckle and rheumatic arthritis, psoriasis, telangiectasis, botryomycosis hominis suppresses spot and forms in the wound healing, atherosclerosis, seborrheic dermatitis and acne; The described inhibition angiogenesis drug of making is used for the purposes of chemotherapy of tumors and/or adjuvant chemotherapy.

Below in conjunction with specific embodiment and experiment, the invention will be further described, and effective site described in the literary composition is the mixture that includes oside compound, and effective monomer is compd A or compd B.

Embodiment 1: the preparation method that has the active ingredient that suppresses angiogenesis function in the Cortex Albiziae

1.1 the 1kg Cortex Albiziae is used 10L, 100 ℃ of reflux, extract, twice of 8L 75% ethanol (methanol of water or variable concentrations or acetone soln) respectively, each 2 hours, filter, remove slag.Merging filtrate, decompression recycling ethanol gets extractum, and this effective site is numbered 298-J.With gained extractum 298-J through D101 type macroporous resin adsorption, water again, 30% ethanol, 80% ethanol, 3 column volumes of each eluting are collected each phase, reclaim 80% ethanol mutually extractum, this effective site is numbered 298-J-D.Gained 298-J-D is separated with 100～200 purpose silica gel in atmosphere pressure, use the volume ratio chloroform: methanol: water is 9: 1: 0.1～6.5: 3.5: 1 gradient elutions, and equal-volume is collected, with the 33rd～44 bottle of merging, reclaim solvent and get extractum, this effective site numbering 298-J-D-G.Gained 298-J-D-G is separated with pressing in the anti-phase styrene post, with 30%～50% acetonitrile gradient eluting, by volume collect (70ml～150ml, 150ml～300ml, 300ml～600ml) as shown in Figure 1, reclaim each phase solvent and get extractum, this effective site is numbered 298-J-D-G-R1 respectively, 298-J-D-G-R2,298-J-D-G-R3.Gained 298-J-D-G-R2 is separated with pressing in the anti-phase C18 chromatographic column, and the methanol aqueous solution eluting with 70% the results are shown in Figure 2, by the unimodal collection of chromatograph, reclaim solvent, lyophilization gets effective monomer 298-J-D-G-R2-A (compd A), 298-J-D-G-R2-B (compd B).Total separation and purification route is seen Fig. 3.

1.2 the HPLC of compd A and compd B analyzes

With gained white powder 298-J-D-G-R2-A, 298-J-D-G-R2-B analyzes through HPLC Agilent 1100 and adopts XDB C-18 post (4.6mm * 150mm, filler φ 5 μ m), and mobile phase adopts 65% methanol (V/V, methanol: water=65: 35).25 ℃ of column temperatures, 215nm detects, sample size 5 μ L.AgilentChemstation software collection, analytical data.The content of 298-J-D-G-R2-A is that the content of 90%298-J-D-G-R2-B is 92.50%.The liquid phase analysis of 298-J-D-G-R2-A the results are shown in Figure 4, and the 298-J-D-G-R2-B liquid phase analysis the results are shown in Figure 5.

1.3: have the active ingredient physical and chemical identification that suppresses angiogenesis function in the Cortex Albiziae

Described active princlple 298-J, 298-J-D, 298-J-D-G, 298-J-D-G-R1,298-J-D-G-R2, the Molish reaction of 298-J-D-G-R3 is aubergine, and the Libermann-Buchard reaction is aubergine, and the Butter of antimony. colour developing is for red.

The MALDI-TOF-MS of active princlple 298-J-D-G shows that quasi-molecular ion peak m/z is at 800-2400.This molecular weight with the Saponin constituents of the bibliographical information of relevant Herba Albiziae is consistent, the results are shown in Figure 6;

This active princlple of above presentation of results all contains at least a above oside compound;

The MALDI-TOF-MS of active princlple 298-J-D-G-R2 shows quasi-molecular ion peak m/z as shown in Figure 7.

Active princlple 298-J-D-G-R2 carries out getting compd A/compd B after anti-phase C18 chromatographic column eluting separates, compd A/compd B, and the Molish reaction is aubergine, and the Libermann-Buchard reaction is aubergine, and the Butter of antimony. colour developing is redness.

1.4: the activity identification of the active princlple that extracts in the Cortex Albiziae

1.4.1 active princlple is to the effect of human microvascular endothelial cell (mvec) inhibition of proliferation

With human microvascular endothelial cell (mvec) (HMEC-1, France national health Institute for Medical Research is so kind as to give) place and contain the 1mmol glutamine, 1 μ g/mL hydrocortisone is cultivated (37 ℃, 5%CO in the MCDB-131 culture fluid of 10ng/mL epidermal growth factor (EGF) and 15% calf serum ₂, 95% humidity).The trophophase cell of taking the logarithm is with 10 ⁵The cell/mL cell inoculation is in 96 orifice plates, and 4 parallel holes are established for every group in 100 μ L/ holes, are positioned over 37 ℃, 5% humidifying CO ₂Incubator spend the night.The 298-J (1～100 μ g/mL) that adds the above extraction separation of variable concentrations, 298-J-D (1～50 μ g/mL), 298-J-D-G (1～10 μ g/mL), 298-J-D-G-R1 (1～10 μ g/mL), 298-J-D-G-R2 (1～10 μ g/mL), 298-J-D-G-R3 (1～10 μ g/mL), 298-J-D-G-R2-A (compd A) (0.05～0.5 μ g/mL), 298-J-D-G-R2-B (compd B) (1～5 μ g/mL), water belongs with yin contrast, cultivate after 72 hours, remove supernatant, every hole adds 10% trichloroacetic acid, 100 μ L lightly to be fixed, and leaves standstill to move on to 4 ℃ behind the 5min and outwell fixative after placing 1h, wash 5 times air drying with deionized water.Every hole adds 37 ℃ of incubators of 0.4% sulphonyl rhodamine B (SRB), 100 μ L and places 30min, washes 5 times air drying with 1% acetate solution.Add the dissolving of 100 μ L 10Mm Tris alkali liquor (pH10.5), spectrophotometer is measured the absorbance (A) of each aperture at 515nm wavelength place automatically.Above-mentioned experiment repeats 3 times.Suppression ratio (%) is calculated as follows:

Result according to gained calculates its half amount of suppression (IC ₅₀), as a result shown in the table 1.

Table 1

The sample title	IC ₅₀(μg/mL)
The sample title	IC ₅₀(μg/mL)	298-J	30.0
298-J-D	10.0	298-J	30.0
298-J-D	10.0	298-J-D-G	2.8
298-J-D-G-R1	1.8	298-J-D-G	2.8
298-J-D-G-R1	1.8	298-J-D-G-R2	1.5
298-J-D-G-R3	1.5	298-J-D-G-R2	1.5
298-J-D-G-R3	1.5	298-J-D-G-R2-A	0.2
298-J-D-G-R2-B	1.2	298-J-D-G-R2-A	0.2

Above presentation of results is along with the propelling of the step of extraction separation, the IC of the effective site of our gained ₅₀Value also reducing, is also progressively improved with regard to the purity that shows effective site.

1.4.2 inhibitory action to the human vascular endothelial transfer ability.

The trophophase cell of taking the logarithm is with 10 ⁵The cell/mL cell inoculation is in 24 orifice plates, and 4 parallel holes are established for every group in the 1mL/ hole, behind the cell attachment, scratch cell along the hole center line, dead cell is removed in the culture medium washing, add water negative control group and 298-J (30.0 μ g/mL) respectively, 298-J-D (10.0 μ g/mL), 298-J-D-G (2.8 μ g/mL), 298-J-D-G-R1 (1.8 μ g/mL), 298-J-D-G-R2 (1.5 μ g/mL), 298-J-D-G-R3 (1.5 μ g/mL), 298-J-D-G-R2-A (0.2 μ g/mL), 298-J-D-G-R2-B (1.2 μ g/mL), respectively at 0h, 6h, 12h, 24h. takes pictures, measure the width of intermediate blank band, record.The width of the blank tape of negative control group cell is less, illustrates that cell migration quantity is higher, and adds after medicine intervened, and the cell migration number obviously reduces.Compd B the results are shown in shown in Figure 8, and the result of other 6 effective sites is to similar with compd A/B, and data do not provide.Above presentation of results effective site, effective monomer all have the effect that suppresses endothelial cell migration.They may suppress the generation of new vessels by the migration that suppresses vascular endothelial cell.

1.4.3 external Matrigel tube method blood vessel inhibition test.

External tubule form the test reference literature [17, ThaloorD, SinghAK, Sidhu GS, et al.Inhibition of angiogenic differenti-ation of human umbilical veinendothelial cells by curcumin[J] .CellGrowth Diff, 1998,9:305-312.] method carry out.Under 4 ℃,, add 50 μ L growth minimizing type Matrigel, pave the back and hatch 4h in 37 ℃ in the bottom of 96 well culture plates.Respectively with the 298-J (1～100 μ g/mL) of variable concentrations, 298-J-D (1～50 μ g/mL), 298-J-D-G (1～10 μ g/mL), 298-J-D-G-R1 (1～10 μ g/mL), 298-J-D-G-R2 (1～10 μ g/mL), 298-J-D-G-R3 (1～10 μ g/mL), 298-J-D-G-R2-A (0.05～0.5 μ g/mL), 298-J-D-G-R2-B (1～5 μ g/mL) is with MCDB-131 culture fluid that contains the 150mL/L calf serum and HMEC-1 (4 * 10 ⁴Cells/well, every pore volume is 0.25mL) join in the culture plate behind the mixing, with the negative contrast of culture medium, after cultivating 24h under 37 ℃, 50mL/L CO2 condition, under inverted microscope, observe the influence that the above internal chrotoplast extracorporeal blood vessel of each sample sample forms.The result shows that above each sample all has the obvious suppression effect to the formation of the external Matrigel tubule of HMEC-1.With the compd B is example, promptly influential to the formation of HMEC-1 tubule when concentration is 1 μ g/ml, the decreased number of tubule; When concentration is 2 μ g/ml, does not almost have after HMEC-1 is adherent and move, do not have tubule substantially and form; Under 3 μ g/ml concentration, the cell attachment difficulty.When the concentration of compd B was 1 μ g/ml, 1.5 μ g/ml, 2 μ g/ml and 3 μ g/ml, compare with the matched group result of other sample of notable difference (the results are shown in shown in Figure 9) of the area that 4 experimental grouies form tube chambers was similar to compd A/B, does not provide.Above presentation of results effective site, effective monomer have the obvious in-vitro suppression angiogenesis function.

1.4.4 albizzia plant extraction suppresses chick chorioallantoic membrane angiogenesis (CAM experiment)

The Rhizoma Euonymus hatching egg, with reference to the method for Zhang Shucheng [18, Zhang Shucheng, Wu Zhikui, Wang Lei, Deng. the application of the chick chorioallantoic membrane test model of research Chinese medicine angiogenic activity and effect. Chinese TCM basis medical journal, 1999,5:16-19.] the preparation chick chorioallantoic membrane.Prepare the microporous filter membrane disk that diameter is 6mm, damp and hot autoclave sterilization with card punch.If the 298-J group, 298-J-D group, 298-J-D-G group, 298-J--D-G-R1 group, 298-J-D-G-R2 group, 298-J-D-G-R3 group, 298-J-D-G-R2-A group, 298-J-D-G-R2-B group and normal saline negative control group.Medicine is dripped on filter paper, make the medicine film, dry up standby.Embryo Gallus domesticus divides into groups after cultivating 3d, the medicine film is affixed on the position of CAM.Behind the dosing 48h, observe medicine film blood vessel number on every side, and photographic recording.Compd B the results are shown in Figure 10.Normal saline negative control group CAM angiogenesis is very obvious, and integral body is cerise, and blood vessel is radial growth under the medicine film, and blood vessel is thick, blood engorgement, and vessel branch is more, and capillary density is big.Compd B administration group has then obviously suppressed angiogenesis, the number of blood vessel of radial growth greatly reduces, blood vessel mainly exists with forms of expression such as fragmentary, mixed and disorderly, vein samples, blood vessel is tiny, sparse, vessel branch is few, and the blood vessel color is light pale yellow, the part rarely has the area vasculosa, perhaps rupture of blood vessel or disappearance around reaching under the medicine diaphragm.The result of other sample is similar to compd A/B, and data do not provide.Above presentation of results effective site, effective monomer have inhibition angiogenesis function in the tangible body.

1.4.5: the molecular structure of Cortex Albiziae effective monomer (compd A, compd B) is identified.

1.4.5.1 compd A is white powder, and is soluble in water, fusing point is 204～204 ℃, and the Molish reaction is aubergine, and the Libermann-Buchard reaction is aubergine, and the Butter of antimony. colour developing is an oside compound for red explanation compd A.Its MALDI-TOF-MS shows quasi-molecular ion peak m/z 2236[M+Na+2] ⁺The results are shown in Figure 11

¹H-NMP (500H _z, C ₃D ₆O, δ ppm) shows 7 unimodal proton signals 0.93 of methyl, 1.02,1.02,1.08,1.15,1.18,1.88,5.58,3 proton signals 3.56 on the oxygen carbon that contain that hang down the field relatively of 1 wide unimodal alkene hydrogen proton signal, 5.21 and 6.31,2 olefinic carbon signals 143.5,123.0 and 1 ester carbonyl group carbon signal 174.6, determine that according to source of students relation and hydrocarbon data the glycoside unit of compd A is the oleanolic acid type triterpene sapogenin, relatively its carbon is composed julibroside III (19.TsuyoshiIkeda, SatokoFujiwara, the KaoruAraki of data and bibliographical information, etal.Cytotoxicglycosides fromAlbiziajulibrissin[J] .J.Nat.Prod, 1997,60 (2): the carbon spectrum data of aglycon acacic acid 102) in full accordly the results are shown in Table 2.

Table 2 glycoside unit acacic acid ¹³C NMR data (py-d ₅)

	¹³C NMR			¹³C NMR
	¹³C NMR			¹³C NMR		NO.	JIII	Compd A	NO.	JIII	Compd A
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15	38.7 26.6 88.5 39.5 55.9 18.5 33.4 39.9 46.9 36.9 23.7 122.9 143.2 41.8 35.7	38.6 26.5 88.4 39.3 56.1 18.7 33.6 39.8 46.9 36.6 23.6 122.9 143.5 41.7 35.6	16 17 18 19 20 21 22 23 24 25 26 27 28 29 30	73.7 51.5 40.8 47.7 35.1 76.7 36.2 27.9 15.7 16.7 17.0 27.1 174.3 29.0 19.0	73.8 51.3 40.6 47.6 35.1 76.9 36.1 27.9 15.6 16.9 17.1 27.0 174.6 28.8 18.9	NO.	JIII	Compd A	NO.	JIII	Compd A

Have 9 saccharide matrix subsignals 4.93 4.87 (1H, d, J=8.0Hz, glcH-1), 5.02 (1H, d, J=8.5Hz, fucH-1), 5.06 (1H, d, J=6.5Hz, xylH-1, (6.03 1H, d, J=8.0Hz, glc ' H-1), (5.88 1H, brs, rha H-1), 6.28 (1H, s, araf H-1), 5.37 (1H, d, J=8.5Hz, glc ' H-1), 4.82 (1H, d, J=6.5Hz, qui H-1), 4.83 (1H, d, J=7.5Hz, qui ' H-1), the bimodal signal 1.76 (3H of methyl proton on 4 desoxy sugars, d, J=5.5Hz, rha6-C H ₁), 1.58 (3H, d, J=5.0Hz, qui ' 6-C H ₃), 1.34 (3H, d, J=6.0Hz, qui6-C H ₃), 1.46 (3H, d, J=6.0Hz, fuc6-C H ₃); 1H-NMR ¹³C-NMR (500H _z, C ₃D ₆O, δ ppm): in methyl carbon signal 17.3,18.6,18.8 and 18.9 on 95.6,99.2,99.4,101.8,103.2,104.5,105.6,106.7 and 111.0 and 4 desoxy sugars of 9 sugared end group carbon signal δ is arranged.Compd A hydrogen spectrum demonstration one typical acetonyl proton signal 8.49 (1H, d, J=8.5Hz); The carbon spectrum has amino-carbon signal δ 58.1 and 1 ethyl carbonyl carbon signal δ 170.1.Illustrating has 9 monosaccharide in the compd A molecule, wherein 4 is 6-deoxidation monosaccharide, and 1 is amino sugar.Except that rha and araf sugar are α-configuration, other sugar are beta comfiguration, and are with the contrast of documentation compound Julibroside III (JIII) sugar moieties carbon spectrum data, in full accord, the replacement kind of compd A and Julibroside III sugar is described, number and connecting mode are the same.The results are shown in Table 3

Table 3 compd A sugar moieties ¹³C NMR and ¹H NMR data

	¹³C NMR ¹H NMR				¹³C NMR ¹H NMR
	¹³C NMR ¹H NMR				¹³C NMR ¹H NMR			JIII	Compd A	Compd A	JIII	Compd A	Compd A
glu 1 2 3 4 5 6 fuc 1 2 3 4 5 6 xyl 1 2 3 4 5 glc ′ 1 2 3 4 5 6 C＝0 CH ₃ qui ′ 1 2 3 4 5 6	106.5 75.5 78.2 71.4 77.3 69.8 103.2 82.1 75.2 72.4 71 17.2 106.8 76.3 77.8 70.6 67 95.5 78.9 76.9 71.6 78.2 62.4 170.1 23.7 99.1 75.3 78.2 76.7 72.4 18.7	106.5 75.4 78.6 71.5 77.1 69.8 103.2 81.7 75.2 72.3 71 17.3 106.7 76.8 77.5 70.8 66.8 95.6 79.1 76.8 71.9 78.6 62.5 170.1 23.7 99.2 75.3 78.3 77.5 72.3 18.8	5.93(d J＝8) 5.01(d，J＝7.5) 1.46(d，J＝6.5) 6.05(d，J＝8.0) 4.83(d 7.8)	rha 1 2 3 4 5 6 glc″ 1 2 3 4 5 6 araf 1 2 3 4 5 qui 1 2 3 4 5 6	101.6 70.3 78.7 84.2 69 18.7 105.5 75 78 71.1 78.2 62.6 110.8 81.9 78.2 85.2 61.8 99.1 75.3 78.2 76.7 72.4 18.7	101.4 70.5 78.6 84.2 68.8 18.6 105.6 75.4 78.6 71.9 78.1 62.5 111.0 81.7 78.1 85.1 61.7 99.4 75.3 78.1 76.8 72.3 18.9	5.88(s) 1.76(d，J＝6.0 5.34(d，J＝9.0) 6.28 4.82(d，J＝7.5) 1.59(d，J＝4.5)	JIII	Compd A	Compd A	JIII	Compd A	Compd A

Except that aglycon and sugared signal, the carbon spectrum has more 20 carbon signals, analyzes its hydrogen spectrum and carbon spectrum signal feature, infers that it may contain two monoterpene structures.In the compd A carbon spectrum δ 145.4 is arranged, 133.6 olefinic carbon and 56.2 methylol signals, illustrate that 9 inboard of monoterpenes are that methylol replaces, can see two groups of triple cutting edges of a knife or a sword 7.09 near 7.0 in the hydrogen spectrum, 7.03, the carbon spectrum respectively has 5 carbon signals 40.8 and 38.7, illustrates that the middle C-6 of MT ' in the outside is the R configuration.Relatively its hydrogen spectrum and carbon spectrum are in full accord with the monoterpene partial data of the chemical compound of bibliographical information JulibrosideIII (JIII).The results are shown in Table 4.Compd A ¹³C-NMR sees Figure 12, ¹H-NMR sees Figure 13.

Table 4. monoterpene acid (MT) ¹³C NMR

MT	¹³C NMR	¹³C NMR	¹H NMR
MT	¹³C NMR	¹³C NMR	¹H NMR	NO.	JIII	Compd A	Compd A
1 2 3 4 5 6 7 8 9 10 MT’ 1 2 3 4 5 6 7 8 9 10	167.4 133.5 145.3 23.4 40.7 79.4 143.8 114.7 56.1 26.6 167.4 127.9 145.3 23.9 40.7 79.7 143.8 114.7 12.8 24.8	167.3 133.6 145.4 23.6 40.8 79.4 143.7 114.7 56.2 26.5 167.3 133.6 145.4 23.6 38.7 79.4 143.7 114.7 12.6 56.2	7.04(t，7.5) 1.79(t，8.5) 6.18(dd，11.0.17.6) 5.16(d，11.5，8a-H) 5.35(d，17.5，8b-H) 1.5(s) 7.04(t，7.5) 1.79(t，8.5) 6.18(dd，11.0.17.6) 5.16(d，11.5，8a-H) 5.35(d，17.5，8b-H) 1.5(s)	NO.	JIII	Compd A	Compd A

IR (KBr) cm-1:3413,2928,1692,1631,1384,1071, the results are shown in Figure 14.

In sum, authenticating compound A is: 3-o-[β-D-xylopyranosyl-(1 → 2)-β-D-pyrans furan glycosyl-(1 → 6)-β-D-2-deoxidation-2-acetamido glucosyl group]-21-o-{ (6s)-2-is trans-and ((6R)-2-trans-2 for 2-methylol-6-methyl-6-o-[4-o-, 6-dimethyl-6-o-β-D-pyrans quinovose base-2,7-octadiene acidic group)-and β-D-pyrans quinovose base]-2,7-octadiene acidic group }-acacic acid-28-o-β-D-glucopyranosyl-(1 → 3)-[α-L-arabinofuranosyl-(1 → 4)]-α-L-rhamnopyranosyl-(1 → 2)-β-D-glucopyranosyl ester promptly (3-o-β-D-xylopyranosyl-(1 → 2)-β-D-fucopyranosyl-(1 → 6)-β-D-2-deoxy-2-acetamidogluco-pyranosyl]-21-o-{ (6s)-2trans-2-hydroxymethyl-6-methyl-6-o-[4-o-((6R)-2-trans-2,6-dimethyl-6-o-β-D-quinovopyranosyl)-2,7-octadienoyl)-and β-D-quinovopyranosyl]-2,7-octadienoyl}-acacic acid-28-o-β-D-glucopyranosyl-(1 → 3)-[α-L-arabinofurano syl-(1 → 4)]-α-L-rhamnopyranosyl-(1 → 2)-β-D-glucopyranosylester

1.5.2 compd B is white powder, and is soluble in water, fusing point is that 204～206 ℃ of Molish reactions are aubergine, and the Libermann-Buchard reaction is aubergine.MALDI-TOF-MS shows quasi-molecular ion peak m/z 2196[M+Na+2] ⁺The results are shown in Figure 15

¹H-NMR (500Hz, C ₃D ₆O, δ ppm) shows 9 unimodal letters 0.96 of methyl proton, 1.03,1.05,1.10 1.16,1.31 and 1.88: the carbon spectrum shows 2 olefinic carbon signals 143.7,123.4, with an ester carbon back signal 174.6, determine that according to source of students relation and hydrocarbon data the glycoside unit of compd B is the oleanane type triterpene sapogenin, compares Julibroside II (the JII) (20.TsuyoshiIkeda that its carbon spectrum data and document are reported for work, SatokoFujiwara, KaoruAraki, etal.Cytotoxicglycosides from AlbiziajulibRissi[J] .J.Nat.Prod, 1997,60 (2): the carbon spectrum data (seeing Table 5) of glycoside unit acacic acid 102), in full accord.

Table 5 glycoside unit ¹³C NMR data (py-d ₅)

	¹³C NMR			¹³C NMR
	¹³C NMR			¹³C NMR		NO.	J II	Compd B	NO.	J II	Compd B
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15	38.7 26.6 88.5 39.5 55.9 18.5 33.4 39.9 46.9 36.9 23.7 122.9 143.2 41.8 35.7	38.6 26.5 88.0 39.3 56.1 18.5 33.4 39.8 46.9 36.8 23.6 123.1 143.4 41.7 35.6	16 17 18 19 20 21 22 23 24 25 26 27 28 29 30	73.7 51.5 40.8 47.7 35.1 76.7 36.2 27.9 15.7 16.7 17.0 27.1 174.3 29.0 19.0	73.5 51.3 40.6 47.5 35.1 76.8 36.1 27.9 15.6 16.8 17.1 27.0 174.6 28.9 18.9	NO.	J II	Compd B	NO.	J II	Compd B

Anomeric proton letter 4.87 (1H, d, J=7.0Hz, quiH-1) 4.82 (1H that nine sugar are arranged, d, J=8.0Hz, qui ' H-1), 4.96 (1H, d, J=8.0Hz, glcH-1), 5.01 (1H, d, J=7.5Hz, fucH-1) 5.09 (1H, d, J=7.0Hz, xylH-1) 5.34 (1H, d, J=8.0Hz, glcH '-1), 5.90 (1H, brs, rhaH '-1), (6.07 1H, d, J=8.0Hz, glc ' H-1), 6.26 (1H, brs, arafH-1), 4 bimodal signal 1.34 (3H of 6-desoxy sugar methyl proton, d, J=6.0Hz, qui6-CH3) 1.58 (3H, d, J=5.0Hz, qui ' 6-CH3) 1.48 (3H, d, J=6.0Hz, 6-CH3), 1.75 (3H, d, J=6.0Hz, 6-CH3). ¹³C-NMR (500H _z, C ₃D ₆O, δ ppm): in observe 9 sugared end group carbon signals 95.5,99.0,99.4,101.4,103.2,105.4,106.5,106.5, the methyl carbon signal 17.2,18.7,18.89 and 18.9 on 107.0 and 110.7,4 desoxy sugars.Illustrating has 9 monosaccharide in the molecule, wherein 4 is the 6-desoxy sugar, and the carbon of Saponin Julibroside II (J II) sugar moieties that compd B sugar moieties carbon spectrum data and document are reported for work is composed data (table 6) and compared, and is in full accord.

Table 6 compd B sugar moieties ¹³C NMR, ¹H NMR data (py-d ₅)

	¹³C NMR ¹H NMR				¹³C NMR ¹H NMR
	¹³C NMR ¹H NMR				¹³C NMR ¹H NMR			J II	Compd B	Compd B	J II	Compd B	Compd B
glu 1 2 3 4 5 6 fuc 1 2 3 4 5 6 xyl 1 2 3 4 5 glc′ 1 2 3 4 5 6 qui′ 1 2 3 4 5 6	106.5 75.5 78.2 71.4 77.3 69.8 103.2 82.1 75.2 72.4 71 17.2 106.8 76.3 77.8 70.6 67 95.5 78.9 76.9 71.6 78.2 62.4 99.1 75.3 78.2 76.7 72.4 18.7	106.5 75.4 78.6 71.5 77.1 69.8 103.2 81.7 75.2 72.3 71 17.1 106.5 76.8 77.5 70.8 66.8 95.5 79.1 76.8 71.9 78.6 62.5 99.4 75.3 78.3 77.5 72.3 18.8	5.90(d J＝8) 5.01(d，J＝7.5) 1.48(d，J＝6.5) 6.07(d，J＝8.0) 4.83(d 7.8)	rha 1 2 3 4 5 6 glc ″ 1 2 3 4 5 6 araf 1 2 3 4 5 qui 1 2 3 4 5 6	101.6 70.3 78.7 84.2 69 18.7 105.5 75 78 71.1 78.2 62.6 110.8 81.9 78.2 85.2 61.8 99.1 75.3 78.2 76.7 72.4 18.7	101.4 70.5 78.6 84.2 68.8 18.9 105.4 75.4 78.6 71.9 78.1 62.5 110.7 81.7 78.1 85.1 61.7 99 75.3 78.1 76.8 72.3 18.89	5.88(s) 1.75(d，J＝6.0 5.34(d，J＝9.0) 6.26 4.82(d，J＝7.5) 1.58(d，J＝4.5)	J II	Compd B	Compd B	J II	Compd B	Compd B

Except that aglycon and sugared signal, the carbon spectrum also has more 20 carbon signals, analyzes its hydrogen spectrum and carbon spectrum signal feature, infers that it may contain two monoterpene structures.In the compd B carbon spectrum δ 144.8 is arranged, 133.5 olefinic carbon and 56.06 methylol signals, illustrate that 9 inboard of monoterpenes are that methylol replaces, can see two groups of triple cutting edges of a knife or a sword 7.09 near 7.0 in the hydrogen spectrum, 7.03, the carbon spectrum respectively has 5 carbon signals 40.8 and 40.8, illustrates that the middle C-6 of MT ' in the outside is the S configuration.Relatively its hydrogen spectrum and carbon spectrum are in full accord with the monoterpene partial data of the chemical compound of bibliographical information Julibroside III (JIII).The results are shown in Table 7, compd B ¹³C-NMR figure sees Figure 16, ¹H-NMR figure the results are shown in Figure 17.

Table 7. monoterpene acid (MT and MT ') ¹³C NMR, ¹H NMR

MT	¹³C NMR ¹H NMR
MT	¹³C NMR ¹H NMR			NO.	J II	Compd B	Compd B
1 2 3 4 5 6 7 8 9 10 MT’ 1 2 3 4 5 6 7 8 9 10	167.4 133.5 145.3 23.4 40.7 79.4 143.8 114.7 56.1 26.6 167.4 127.9 145.3 23.9 40.7 79.7 143.8 114.7 12.8 24.8	167.3 133.6 145.4 23.6 40.8 79.4 143.7 114.7 56.2 26.5 167.3 133.6 145.4 23.6 40.8 79.4 143.7 114.7 12.7 24.7	7.05(t，7.5) 1.79(t，8.5) 6.18(dd，11.0.17.6) 5.16(d，11.5，8a-H) 5.36(d，17.5，8b-H) 1.5(s) 7.04(t，7.5) 1.79(t，8.5) 6.16(dd，11.0.17.6) 5.16(d，11.5，8a-H) 5.35(d，17.5，8b-H) 1.5(s)	NO.	J II	Compd B	Compd B

IR (KBr) cm-1:3413,2926,1694,1644,1374,1076 the results are shown in Figure 18

Authenticating compound B is in sum:

Chemical name 3-o-[β-D-xylopyranosyl-(1 → 2)-β-D-pyrans furan glycosyl-(1 → 6)-β-D-glucopyranosyl]-21-o-{ (6s)-2-is trans-and ((6R)-2-trans-2 for 2-methylol-6-methyl-6-o-[4-o-, 6-dimethyl-6-o-(β-D-pyrans quinovose base)-2,7-octadiene acidic group)-and β-D-pyrans quinovose base]-2,7-octadiene acidic group]-acacic acid-28-o-β-D-pyrans Fructus Vitis viniferae-(1 → 3)-[α-L-arabinofuranosyl-(1 → 4)]-α-L-rhamnopyranosyl-(1 → 2)-β-D-glucopyranosyl ester is 3-o-[β-D-xylopyranosyl-(1 → 2)-β-D-fucopyranosyl-(1 → 6)-β-D-gluCopyranosyl]-21-o-{ (6s)-2-trans-2-hydroxymethyl-6-methyl-6-o-[4-o-((6R)-2-trans-2,6-dimethyl-6-o-β-D-quinovopyranosyl)-2,7-octadienoyl)-and β-D-quinovopyranosyl]-2,7-octadienoyl}-acacic acid-28-o-β-D-glucopyranosyl-(1 → 3)-[α-L-arabinofuranosyl-(1 → 4)]-α-L-rhamnopyranosyl-(1 → 2)-β-D-glucopyranosyl ester.

The present invention is described in conjunction with most preferred embodiment, yet after having read foregoing of the present invention, those skilled in the art can understand and do many changes also can obtain identical or similar result in disclosed embodiment, and do not exceed design of the present invention, spirit and scope.More particularly, obviously some chemistry obtains identical or similar result with the alternative reagent disclosed herein of physiological related reagent.All similarly replace and modify for a person skilled in the art, obviously think all in spirit of the present invention, scope and design and the claim scope that promptly all above-mentioned these equivalent form of values all fall within claims of the present invention institute restricted portion equally.

Claims

1, having the active ingredient that suppresses angiogenesis function in the Cortex Albiziae is oside compound.

2, according to having the active ingredient that suppresses angiogenesis function in the described Cortex Albiziae of claim 1, it is characterized in that: described oside compound wherein contains compd A and/or compd B, being characterized as of the molecular structure of described compd A:

(I) R wherein ₁: CH ₃R ₂: NHAC R ₃: OH C-6:R

The molecular formula of compd A is: C ₁₀₄H ₁₆₅O ₄₉N, molecular weight is 2211, fusing point is 202～204 ℃, chemistry is by name: 3-o-[β-D-xylopyranosyl-(1 → 2)-β-D-pyrans furan glycosyl-(1 → 6)-β-D-2-deoxidation-2-acetamido glucosyl group]-21-o-{ (6s)-2-is trans-and ((6R)-2-trans-2 for 2-methylol-6-methyl-6-o-[4-o-, 6-dimethyl-6-o-β-D-pyrans quinovose base-2,7-octadiene acidic group)-and β-D-pyrans quinovose base]-2,7-octadiene acidic group }-acacic acid-28-o-β-D-glucopyranosyl-(1 → 3)-[α-L-arabinofuranosyl-(1 → 4)]-α-L-rhamnopyranosyl-(1 → 2)-β-D-glucopyranosyl ester

Being characterized as of the molecular structure of described compd B:

(II) R wherein ₁: CH ₃R ₂: OH C-6:S

3, the preparation method that has the active ingredient that suppresses angiogenesis function in the Cortex Albiziae, it is characterized in that: with dry bark, peel of stem, leaf, flower, seed, soybean pod or the root of organic solvent, water-containing organic solvent or water extraction Cortex Albiziae, and with extracting liquid filtering, be condensed into extractum; The extractum employing D-101 type macroporous resin eluting of gained separates, and collects active princlple and carries out the separation of silica gel column chromatography eluting, regathers active princlple and carries out anti-phase styrene chromatographic column separation, gets active princlple, reclaims solvent and becomes extractum, and dry powder is made in lyophilization.

4, according to the preparation method that has the active ingredient that suppresses angiogenesis function in the described Cortex Albiziae of claim 3, it is characterized in that: separate described anti-phase styrene chromatographic column to such an extent that active princlple carries out getting oside compound A and oside compound B after anti-phase C18 chromatographic column eluting separates.

5, have the application of the active ingredient that suppresses angiogenesis function in the Cortex Albiziae, it is characterized in that: described oside compound has antineoplastic vascular to generate and the purposes of other and associated angiogenesis disease treatment, prevention; Or in the medicine that suppresses angiogenesis, use; Perhaps as conventional pharmaceutical carrier and/or excipient; The perhaps application in the health food of preparation inhibition angiogenesis; The perhaps application in the cosmetics of preparation inhibition angiogenesis.

6, according to the application that has the active ingredient that suppresses angiogenesis function in the described Cortex Albiziae of claim 5, it is characterized in that: when making described inhibition angiogenesis drug, can add one or more pharmaceutically acceptable carriers.

7, according to the application that has the active ingredient that suppresses angiogenesis function in the described Cortex Albiziae of claim 6, it is characterized in that: described carrier comprises the diluent of pharmaceutical field routine, excipient, filler, binding agent, wetting agent, disintegrating agent, absorption enhancer, surfactant, absorption carrier, lubricant, emulsifying agent, osmotic pressure regulator can also add flavouring agent, sweeting agent etc. in case of necessity.

8, according to the application that has the active ingredient that suppresses angiogenesis function in the described Cortex Albiziae of claim 6, it is characterized in that: when making described inhibition angiogenesis drug, preparation process that can be routinely, use separately or prepare the medicine of operable various different dosage forms clinically, for example various ways such as injection, tablet, powder, granule, capsule, oral liquid, unguentum, cream or Emulsion with other medicines.

9, according to the application that has the active ingredient that suppresses angiogenesis function in the described Cortex Albiziae of claim 6, it is characterized in that: the described inhibition angiogenesis drug of making is used to prevent and/or treat at least a disease that is selected from one of the following: cancer and/or cancer metastasis, hemangioma, fibrohemangioma, neovascular glaucoma, diabetic renal papillary necrosis, retinopathy of prematurity and retinal vein occlusion, retinal degeneration, retrolental fibroplasia, the keratopathy that trachomatous conjunctivitis, angiogenesis cause, climacteric macula lutea and degeneration of macula, psoriasis, old plate-like speckle degeneration and rheumatic arthritis, psoriasis, telangiectasis, botryomycosis hominis, suppress spot in the wound healing and form atherosclerosis, seborrheic dermatitis and acne.

10, according to the application that has the active ingredient that suppresses angiogenesis function in the described Cortex Albiziae of claim 4, it is characterized in that: the described inhibition angiogenesis drug of making is used for the purposes of chemotherapy of tumors and/or adjuvant chemotherapy.