CN105152873A - New lignan compounds, and preparation method and medical application thereof - Google Patents

New lignan compounds, and preparation method and medical application thereof Download PDF

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CN105152873A
CN105152873A CN201510672462.2A CN201510672462A CN105152873A CN 105152873 A CN105152873 A CN 105152873A CN 201510672462 A CN201510672462 A CN 201510672462A CN 105152873 A CN105152873 A CN 105152873A
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叶澄
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Wenzhou Hongchengxiang Technology Co Ltd
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C41/00—Preparation of ethers; Preparation of compounds having groups, groups or groups
    • C07C41/01—Preparation of ethers
    • C07C41/34—Separation; Purification; Stabilisation; Use of additives
    • C07C41/36—Separation; Purification; Stabilisation; Use of additives by solid-liquid treatment; by chemisorption
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C43/00—Ethers; Compounds having groups, groups or groups
    • C07C43/02—Ethers
    • C07C43/20—Ethers having an ether-oxygen atom bound to a carbon atom of a six-membered aromatic ring
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Abstract

The invention discloses new lignan compounds, and a preparation method and medical application thereof. The lignan compounds are reported for the first time, have novel structure, and can be extracted, separated and purified from dry bark of Mangnolia officinalis. The in-vitro test proves that the compounds have a protective action on H2O2 oxidation damaged ECV-304 cells. The determination on cell supernatant MDA (malondialdehyde) content, SOD (superoxide dismutase) activity and GSH-Px (glutathione peroxidase) activity proves that the compounds (I) have the antioxidation characteristic of removing free radicals and active oxygen and can be used for developing vessel-protecting drugs.

Description

A kind of new Lignanoids compounds and preparation method thereof and medicinal use
Technical field
The invention belongs to technical field of pharmaceuticals, be specifically related to from the dry bark of the bark of official magnolia, be separated obtain a kind of and there is Lignanoids compounds of vascular protection effect and preparation method thereof.
Background technology
The bark of official magnolia, derive from the dry dry hide of Magnoliaceae magnolia bark of official magnolia MagnoliaofficinalisRehdetWils. and Magnolia bilola MagnoliaofficinalisRehd.etWils.var.bilobaRehd.etWils., branch skin and Gen Pi, main product in Sichuan, Hubei, Zhejiang, the ground such as Fujian and Anhui.Begin to be loaded in Shennong's Herbal, be classified as middle product.The bark of official magnolia is conventional Chinese medicine, have remove dampness by means of aromatics, promoting the circulation of qi disappears long-pending, eliminating dampness except effect that is full, lowering the adverse QI to relieve asthma, the illness such as cure mainly stagnation of QI due to dyspepsia, abdominal distention constipation, retention of dampness in middle-JIAO, institute ruffian Tu Wan, phlegm stops up the circulation of vital energy in the wrong direction, fullness sensation in chest is breathed with cough.The bark of official magnolia is had to be used as medicine in many conventional prescriptions and Chinese patent medicine preparation.Modern times grind to make internal disorder or usurp and show, main in the bark of official magnolia have good activity containing lignanoid, alkaloid and volatilization wet goods composition to Digestive tract, and have antibacterial, CNS inhibition, flaccidity and the effect such as antitumor.
In successive dynasties main book on Chinese herbal medicine, contained its medicinal part of the certified products bark of official magnolia is bark, the record of bark of official magnolia fruit (abundant) is had in " Mingyi Bielu " and Compendium of Material Medica, there is the record of its bud (Flos Magnoliae Officinalis) in " medicine materical crude slice is newly joined ", in " Chinese Pharmacopoeia " 2010 editions one, recorded bark of official magnolia magnolia obovata flower standard.But be showed no the medicinal record of Leaf of Magnolia officinalis at all times in pharmacy monograph, only have certain application among the people.
All prove from cardiovascular physiology, pharmacology, Cytobiology and molecular biology etc. are multi-field, blood vessel plays important self-regulation effect to blood pressure.Antiotasis is the important factor determining blood pressure, and hypertension is mainly because caused by the increase of blood vessel total peripheral resistance.Antiotasis is by the adjustment of numerous endogenous vaso-active substance, Leaf of Magnolia officinalis the different extracted parts shows the regulating effect to antiotasis, prompting Leaf of Magnolia officinalis can further grind as regulation of blood vessels medicine the exploitation that makes internal disorder or usurp, especially it should be noted that the vasodilatory effect of its large polar fraction, show that Leaf of Magnolia officinalis has potential hypotensive activity, therefore, whether can have on the basis of hypotensive activity in body grinding further the Leaf of Magnolia officinalis extract that makes internal disorder or usurp, can specify Leaf of Magnolia officinalis carry out sending out utilization as the hypertensive vasodilator for the treatment of.
Summary of the invention
The object of this invention is to provide and a kind ofly from the dry bark of the bark of official magnolia, be separated obtain a kind of there is Lignanoids compounds of vascular protection effect and preparation method thereof.
Above-mentioned purpose of the present invention is achieved by technical scheme below:
There is the compound (I) of following structural formula,
The preparation method of described compound (I), comprise following operation steps: the dry bark of the bark of official magnolia is pulverized by (a), extract with 80 ~ 90% alcohol heat reflux, united extraction liquid, be concentrated into without alcohol taste, use sherwood oil, ethyl acetate and water saturated n-butanol extraction successively, obtain petroleum ether extract, acetic acid ethyl ester extract and n-butyl alcohol extract respectively; Acetic acid ethyl ester extract macroporous resin removal of impurities in (b) step (a), first use 15% ethanol elution, 6 column volumes, then use 75% ethanol elution, 10 column volumes, collect 75% ethanol eluate, concentrating under reduced pressure obtains 75% ethanol elution thing medicinal extract; C in () step (b), 75% ethanol elution medicinal extract purification on normal-phase silica gel is separated, obtain 5 components successively with the methylene chloride-methanol gradient elution that volume ratio is 80:1,50:1,30:1,15:1 and 1:1; D in () step (c), component 4 is separated further by purification on normal-phase silica gel, obtain 3 components successively with the methylene chloride-methanol gradient elution that volume ratio is 25:1,15:1 and 8:1; E in () step (d), component 2 reverse phase silica gel of octadecylsilane bonding is separated, with the methanol aqueous solution isocratic elution that concentration expressed in percentage by volume is 70%, collect 10 ~ 12 column volume elutriants, elutriant concentrating under reduced pressure obtains pure compound (I).
Described macroporous resin is AB-8 type macroporous adsorbent resin.
It is 85% that described alcohol heat reflux extracts the alcohol concn adopted.
A kind of pharmaceutical composition, the described compound (I) wherein containing treatment significant quantity and pharmaceutically acceptable carrier.
Described compound (I) application in the medicine preparing vascular protection.
The application of described pharmaceutical composition in the medicine preparing vascular protection.
When the compounds of this invention is used as medicine, directly can uses, or use with the form of pharmaceutical composition.
This pharmaceutical composition contains the compounds of this invention (I) for the treatment of significant quantity, and all the other are acceptable on pharmacology, nontoxic to humans and animals and pharmaceutically acceptable carrier of inertia and/or vehicle.
Described pharmaceutically acceptable carrier or vehicle are that one or more are selected from solid, semisolid and liquid diluent, filler and pharmaceutical preparation assistant agent.Pharmaceutical composition of the present invention is used with the form of per weight dose.Medicine of the present invention is applied to by form that is oral or injection the patient needing treatment.For time oral, tablet, slow releasing tablet, controlled release tablet, capsule, dripping pill, micropill, suspensoid, emulsion, powder or granule, oral liquid etc. can be made into; During for injecting, can be made into water-based or oily solution, aseptic powder injection, liposome or the emulsion etc. of sterilizing.
Figure of description
Fig. 1 is compound (I) structural formula;
Fig. 2 is that the theoretical ECD value of compound (I) compares with experiment ECD value.
Embodiment
Further illustrate essentiality content of the present invention below in conjunction with embodiment, but do not limit scope with this.Although be explained in detail the present invention with reference to preferred embodiment, those of ordinary skill in the art should be appreciated that and can modify to technical scheme of the present invention or equivalent replacement, and does not depart from essence and the scope of technical solution of the present invention.
Embodiment 1: compound (I) is separated preparation and structural identification
Reagent source: ethanol, sherwood oil, ethyl acetate, propyl carbinol, methylene dichloride are analytical pure, purchased from Shanghai Ling Feng chemical reagent company limited, methyl alcohol, analytical pure, purchased from Jiangsu Han Bang chemical reagent company limited.
Preparation method: the dry bark (8kg) of the bark of official magnolia is pulverized by (a), (25L × 3 time) are extracted with 85% alcohol heat reflux, united extraction liquid, be concentrated into without alcohol taste (6L), use sherwood oil (6L × 3 time), ethyl acetate (6L × 3 time) and water saturated propyl carbinol (6L × 3 time) to extract successively, obtain petroleum ether extract, acetic acid ethyl ester extract (355g) and n-butyl alcohol extract respectively; Acetic acid ethyl ester extract AB-8 type macroporous resin removal of impurities in (b) step (a), first use 15% ethanol elution, 6 column volumes, use 75% ethanol elution, 10 column volumes again, collect 75% ethanol eluate, concentrating under reduced pressure obtains 75% ethanol elution thing medicinal extract (136g); C in () step (b), 75% ethanol elution medicinal extract purification on normal-phase silica gel is separated, successively with volume ratio be 80:1 (8 column volumes), the methylene chloride-methanol gradient elution of 50:1 (8 column volumes), 30:1 (6 column volumes), 15:1 (8 column volumes) and 1:1 (5 column volumes) obtains 5 components; D component 4 (31g) is separated further by purification on normal-phase silica gel in () step (c), successively with volume ratio be 25:1 (8 column volumes), the methylene chloride-methanol gradient elution of 15:1 (10 column volumes) and 8:1 (6 column volumes) obtains 3 components; E in () step (d), component 2 (11g) reverse phase silica gel of octadecylsilane bonding is separated, with the methanol aqueous solution isocratic elution that concentration expressed in percentage by volume is 70%, collect 10-12 column volume elutriant, elutriant concentrating under reduced pressure obtains pure compound (I) (28mg).
Structural identification: HR-ESIMS shows [M+Na] +for m/z431.1716, can obtain molecular formula in conjunction with nuclear-magnetism feature is C 21h 28o 8, degree of unsaturation is 8.Hydrogen nuclear magnetic resonance modal data δ h(ppm, DMSO-d 6, 500MHz): H-2 (6.10, d, J=2.0), H-5 (6.50, d, J=8.0), H-6 (6.25, dd, J=8.0, 2.0), H-7 (2.33, dd, J=13.5, 4.5), H-7 (1.94, dd, J=13.5, 4.5), H-8 (1.85, dt, J=8.0, 3.5), H-9 (4.57, d, J=1.0), H-2 ' (6.62, d, J=1.5), H-5 ' (6.62, d, J=8.0), H-6 ' (6.57, d, J=8.0, 1.5), H-7 ' (4.34, d, J=10.0), H-8 ' (2.15, m), H-9 ' (4.09, t, J=8.5), H-9 ' (3.89, dd, J=8.5, 6.5), 3-OCH 3(3.74, s), 3 '-OCH 3(3.73, s), 9-OCH 3(3.09, s), carbon-13 nmr spectra data δ c(ppm, DMSO-d 6, 125Hz): 130.1 (C, 1-C), 110.5 (CH, 2-C), 146.8 (C, 3-C), 143.5 (C, 4-C), 113.7 (CH, 5-C), 120.1 (CH, 6-C), 37.8 (CH 2, 7-C), 49.2 (CH, 8-C), 108.5 (CH, 9-C), 134.7 (C, 1 '-C), 109.1 (CH, 2 '-C), 146.7 (C, 3 '-C), 145.2 (C, 4 '-C), 113.7 (CH, 5 '-C), 118.5 (CH, 6 '-C), 75.8 (CH, 7 '-C), 49.9 (CH, 8 '-C), 69.4 (CH 2, 9 '-C), 53.8 (CH 3, 3-OCH 3), 53.7 (CH 3, 3 '-OCH 3), 52.4 (CH 3, 9-OCH 3), carbon atom mark is see Fig. 1. 1this compound of HNMR spectrum display exists two 1, 3, 4-tri-substituted aroma ring [δ H6.10 (1H, d, J=2.0Hz, H-2), 6.50 (1H, d, J=8.0Hz, H-5) and 6.25 (1H, dd, J=8.0, 2.0Hz, and 6.62 (1H H-6), d, J=1.5Hz, H-2 '), 6.62 (1H, d, J=8.0Hz, H-5 '), with 6.57 (1H, dd, J=8.0, 1.5Hz, H-6 ')], two containing oxygen methyne [δ H4.57 (1H, d, J=1.0Hz, H-9) and 4.34 (1H, d, J=10.0Hz, H-7 ')], three methoxyl group [δ H3.74 (3H, s, 3-OCH 3), 3.73 (3H, s, 3 '-OCH 3), and 3.09 (3H, s, 9-OCH 3)], one containing Oxymethylene [δ H4.09 (1H, t, J=8.5Hz, H-9 ' a) He 3.89 (1H, dd, J=8.5,6.5, H-9 ' is b)), methylene radical [δ H2.33 (1H, dd, J=13.5,4.5, H-7a) and 1.94 (1H, dd, J=13.5,4.5, ], and two methynes [δ H1.85 (1H, dt, J=8.0 H-7b), 3.5, H-8) and 2.15 (1H, m, H-8 ')]. 13cNMR spectrum shows 21 carbon signals, comprising 12 aromatic carbons of two aromatic rings, two containing oxygen methyne [δ C108.5 (C-9) and 75.8 (C-7 ')], one containing Oxymethylene [δ C69.4 (C-9 ')], three methoxyl group [δ C53.8 (3-OCH 3), 53.7 (3 '-OCH 3) and 52.4 (9-OCH 3)], two methynes [δ C49.9 (C-8 ') and 49.2 (C-8)], and a methylene radical [δ C37.8 (C-7)].H-7 and H-9 and H-7 ' and H-8 and H-8 ' and H-9 '-OCH during coupling constant (1.0Hz) less between H-8 and H-9 and NOESY compose 3dependency show that the absolute structure of C-8 and C-8 ' is respectively S and R.In addition C-7 ' be configured as R.Comprehensive hydrogen spectrum, carbon spectrum, HMBC spectrum and NOESY spectrum, and document is about correlation type nuclear magnetic data, can substantially determine this compound as shown in Figure 1, steric configuration is determined further by ECD test, theoretical value and experimental value basically identical (Fig. 2).
Embodiment 2: compound (I) pharmacological action is tested
One, material and instrument
Vascular endothelial cell (ECV-304) is presented by immunity teaching and research room of medical college of Shandong University.Compound (I) is made by oneself, and preparation method is shown in that embodiment 1, HPLC normalization method purity is greater than 98%.RPMI-1640 dehydrated medium, trypsinase are purchased from Gibeo company of the U.S..Foetal calf serum is purchased from Tianjin TBD company.MTT, agarose, AnnexinV-FITC are purchased from Sigma Co., USA.LDH, MDA, SOD, GSH-Px mensuration test kit is purchased from Nanjing and builds up Bioengineering Research Institute.DMSO is purchased from Chinese Medicine (group) Solution on Chemical Reagents in Shanghai company limited.Hematorylin is purchased from Foochow and steps true tumor Technology Co., Ltd..Rhodamine123 is purchased from Solarbic company of the U.S..Positive drug Ligustrazine is purchased from Yuan Ye bio tech ltd, Shanghai.
Inverted light microscope (Japanese OlymPus company), CO2gas incubator (FormaScientific company of the U.S.), electronic analytical balance (ER-182A type) (Japanese A & D company), Bechtop (ZHJH-1209 type) (Shanghai ZHICHENG Anaiytical Instrument Manufacturing Co., Ltd.), flow cytometer (FACSVantage type) (BeetonDiehnson company of the U.S.), constant temperature oscillator (Taicang, Jiangsu experimental installation factory), 1420Vietor 3multiple labeling analyser (PerkinElmerLifescience company of the U.S.), the visible ultraviolet grating spectrophotometer of 721 type (Shanghai exact science company limited), electric heating constant-temperature water-bath tank (production of Shanghai Medical instrument three factory), enzyme-linked immunosorbent assay instrument (MK3Multiskan) (Shanghai ThermoLabsystem analytical instrument company).
Two, test method
1, cell cultures
ECV-304 cell is cultivated based on 37 DEG C with RPMI-1640,5%CO 2secondary Culture in incubator.Microscopic observation, goes down to posterity with tryptic digestion when cell confluency rate reaches more than 70%, prepares cell suspension to grow stable 3-5 for cell.
2, cell grouping and process
Get well-grown ECV-304 cell and make cell suspension, be inoculated in 96 holes, 24 holes, 6 porocyte culture plates or culture dish, 37 DEG C, 5%CO 2cultivate 24h.Be divided into six groups at random: 1. normal group; 2. H 2o 2model group (150 μm of ol/L); 3. Ligustrazine (TMP) (50 μm of ol/L)+H 2o 2group; 4. compound (I) (10 μm of ol/L)+H 2o 2group; 5. compound (I) (50 μm of ol/L)+H 2o 2group; 6. compound (I) (100 μm of ol/L)+H 2o 2group.
3, experimental project and Testing index
3.1MTT method detects cell viability
Well-grown ECV-304 cell is prepared into 5 × 10 4the cell suspension of/mL, is inoculated in 96 orifice plates by every hole 100 μ L, puts 37 DEG C, 5%CO 2hatch 24h.Cell divides into groups and processes.After continuing to cultivate 6h and 12h, every hole adds 10 μ LMTT solution (final concentration 0.5mg/L) and puts 37 DEG C, 5%CO 2after incubator hatches 4h, every hole adds 100 μ LDMSO, and the built-in enzyme-linked immunosorbent assay instrument of 60s, 30min that vibrates after leaving standstill 10min detects 570nm place absorbance (OD 570).
By formula:
Cell injury inhibiting rate=(medication group OD 570-model group OD 570)/(normal group OD 570-model group OD 570) × 100%, calculates cell injury inhibiting rate.
3.2LDR release rate assay
Get well-grown ECV-304 cell and make l × 10 5the cell suspension inoculation of/mL, in 24 orifice plates, puts 37 DEG C, 5%CO 2hatch 24h.Cell divides into groups and processes the same.Continue to cultivate 6h and 12h, collect culture supernatant.After PBS washs 2 times, every hole adds 0.5mL cell pyrolysis liquid (150mmol/LNaCl, 150mmol/LTris-HCI, lmmol/LEDTA, 1%TritonX-100), vibrate after 4 DEG C of standing 15min several minutes, 10000rpm, 4 DEG C of centrifugal 10min collecting cell lysates, measure the LDH activity in supernatant liquor and cell pyrolysis liquid respectively with reference to LDH test kit specification sheets.
By formula: LDH activity in LDH release rate=supernatant liquor/(in supernatant liquor in LDH activity+cell pyrolysis liquid LDH activity) × 100%, calculates LDH release rate.
3.3 enzyme biochemical process measures MDA content, SOD vigor, GSH-Px vigor
Get well-grown ECV-304 cell and make 1 × 10 5the cell suspension inoculation of/mL, in 24 orifice plates, puts 37 DEG C, 5%CO 2hatch 24h.Cell divides into groups and processes the same.Continue to cultivate 12h, collect culture supernatant.Cell MDA content, SOD vigor, GSH-Px vigor is measured by MDA, SOD, GSH-Px detection kit specification sheets.
4, data processing
The statistical software that MicrosoftExcel carries processes, experimental data with represent, group difference t checks.
Three, result and conclusion
1, compound (I) is to H 2o 2the impact of damage ECV-304 cells survival rate
Produce succinodehydrogenase in normal live cells mitochondrial process, faint yellow MTT can be reduced into the cured crystallization of water-fast hepatic first, crystallization quantity is directly proportional to viable count.Result: ECV-304 cell is through H 2o 2after (150 μm of ol/L) oxidative damage 6h and 12h, cell OD value obviously reduces and have statistical significance (P<0.01) compared with normal group, shows that cell viability declines.And compound (I) has provide protection to cell, significantly H can be suppressed 2o 2to the oxidative damage of cell, improve survival rate.Effect 6h, 50,100 μm of ol/L compounds (I) are respectively 16.67% (P<0.05) and 28.21% (P<0.01) to cell injury inhibiting rate.Effect 12h, 10,50,100 μm of ol/L compounds (I) rise to 24.85% (P<0.05) to cell injury inhibiting rate, 29.64% (P<0.01) and 38.47% (P<0.01).In table 1 (* * P<0.01vsNormal; #p<0.05, #p<0.01vsH 2o 2group; ▲p<0.05, ▲ ▲p<0.01vsTMPgroup, lower same).
2, compound (I) is to H 2o 2the impact of damage ECV-304 cell LDH release rate
Serum lactic dehydrogenase energy catalysis lactic acid generates pyruvic acid, and pyruvic acid and 2,4 dinitrophenyl hydrazine react and generate pyruvic acid dinitrophenylhydrazone, in red-brown in basic solution, indirectly can obtain Ldh Activity by colorimetric estimation reaction product.Result shows: compared with normal group, and model group ECV-304 cell LDH release rate significantly increases (P<0.01).With model group ratio, compound (I) respectively group cell LDH release rate all reduces: 10,50,100 μm of ol/L compound (I) effect 6h, and the LDH release rate of cell reduces to 20.54% (P<0.01) and 21.22% (P<0.01); 50,100 μm of ol/L compound (I) effect 12h, the LDH release rate of cell reduces to 33.64% (P<0.01) and 29.53% (P<0.01).Compound (I), with the increase of concentration, also strengthens gradually the provide protection of oxidative damage ECV-304 cell, presents certain dose-effect relationship.In table 2.
3, compound (I) is to H 2o 2the impact of damage ECV-304 cell MDA content, SOD, GSH-Px vigor
Experimental result shows: significantly increase (P<0.01) than MDA content in model group ECV-304 cell culture fluid with normal group.Compared with model group, 10,50,100 μm of ol/L compound (I) group cell MDA growing amounts all obviously reduce, be respectively 3.00 ± 0.79nmol/mL (P<0.05), 2.86 ± 0.75nmol/mL (P<0.05) and 2.69 ± 0.45nmol/mL (P<0.01).Contrast display between compound (I) each concentration group group, with the increase of concentration, the provide protection of compound (I) to ECV-304 cytolipin peroxide injury also strengthens gradually, presents certain dose-effect relationship.The results are shown in Table 3.
Experimental result shows: compared with normal group, and model group ECV-304 cell SOD is active significantly reduces (P<0.01).Compared with model group, 50,100 μm of ol/L compounds (I) all can to strengthen the SOD of ECV-304 cell active, SOD activity rises to 17.9 ± l.34U/mL (P<0.05) and 19.25 ± 0.81U/mL (P<0.01) respectively.And with the increase of concentration, the SOD activity of cell also improves gradually, shows that compound (I) can strengthen the antioxygen free action of ECV-304 cell, a certain amount of effect relationship of tool; 10 μm of ol/L compounds (I), though can improve SOD activity, do not have remarkable statistical significance yet.In table 3.
Experimental result shows: compared with normal group, and in model group ECV-304 cell culture fluid, GSH-Px is active significantly reduces (P<0.01).Compared with model group, 10,50,100 μm of ol/L compound (I) group cell GSH-Px activity are significantly increased, be respectively 73.8 ± 10.3U/mL (P<0.05), 92.2 ± 8.5U/mL (P<0.01) and 102.5 ± 10.3U/mL (P<0.01).And with the increase of compound (I) concentration, the GSH-Px activity of ECV-304 cell also improves gradually, shows that the ability of cellular anti-oxidant effect also strengthens gradually, presents certain dose-effect relationship.The results are shown in Table 3.
Sum up: this experiment adopts H 2o 2as exogenous free radical generation system, can induction of vascular endothelial cell (ECV-304)) oxidativestress damage, promote apoptosis.Detected by mtt assay and LDH activity and confirm that compound (I) is to H 2o 2oxidative damage cell has provide protection; By cell conditioned medium liquid MDA content, SOD is active, GSH-Px is active mensuration, confirm that compound (I) has the anti-oxidation characteristics of scavenging free radicals and active oxygen.
Table 1 compound (I) is to H 2o 2the impact of damage ECV-304 cells survival rate ( n=8)
Table 2 compound (I) is to H 2o 2the impact of damage ECV-304 cell LDH release rate ( n=8)
Table 3 compound (I) is to H 2o 2the impact of damage ECV-304 cell MDA content, SOD, GSH-Px vigor ( n=8)
Embodiment 3
The preparation of tablet: by embodiment 1 method first obtained compound (I), and the salt utilizing organic acid to make as tartrate or citric acid or formic acid or oxalic acid etc., mineral acid example hydrochloric acid or sulfuric acid or phosphoric acid, vehicle is added, pelletizing press sheet than the ratio for 1:9 in itself and excipient weight.
Embodiment 4
Prepared by oral liquid: by embodiment 1 method first obtained compound (I), and the salt utilizing organic acid to make as tartrate or citric acid or formic acid or oxalic acid etc., mineral acid example hydrochloric acid or sulfuric acid or phosphoric acid, oral liquid method for making makes oral liquid routinely.
Embodiment 5
The preparation of capsule or granule: by embodiment 1 method first obtained compound (I), and the salt utilizing organic acid to make as tartrate or citric acid or formic acid or oxalic acid etc., mineral acid example hydrochloric acid or sulfuric acid or phosphoric acid, add vehicle in itself and excipient weight than the ratio for 1:9, make capsule or granule.
Embodiment 6
The preparation of injection liquid: by embodiment 1 method first obtained compound (I), and the salt utilizing organic acid to make as tartrate or citric acid or formic acid or oxalic acid etc., mineral acid example hydrochloric acid or sulfuric acid or phosphoric acid, inject with water routinely, essence filter, injection liquid is made in embedding sterilizing.
Embodiment 7
The preparation of aseptic powder injection: by embodiment 1 method first obtained compound (I), and the salt utilizing organic acid to make as tartrate or citric acid or formic acid or oxalic acid etc., mineral acid example hydrochloric acid or sulfuric acid or phosphoric acid, be dissolved in sterile water for injection, stirring makes molten, filter with aseptic suction funnel, aseptic essence filter again, be sub-packed in ampoule, after frozen drying, aseptic sealing by fusing obtains powder injection.
The effect of above-described embodiment is essentiality content of the present invention is described, but does not limit protection scope of the present invention with this.Those of ordinary skill in the art should be appreciated that and can modify to technical scheme of the present invention or equivalent replacement, and does not depart from essence and the protection domain of technical solution of the present invention.

Claims (7)

1. there is the compound (I) of following structural formula,
2. the preparation method of compound according to claim 1 (I), it is characterized in that comprising following operation steps: the dry bark of the bark of official magnolia is pulverized by (a), extract with 80 ~ 90% alcohol heat reflux, united extraction liquid, be concentrated into without alcohol taste, use sherwood oil, ethyl acetate and water saturated n-butanol extraction successively, obtain petroleum ether extract, acetic acid ethyl ester extract and n-butyl alcohol extract respectively; Acetic acid ethyl ester extract macroporous resin removal of impurities in (b) step (a), first use 15% ethanol elution, 6 column volumes, then use 75% ethanol elution, 10 column volumes, collect 75% ethanol eluate, concentrating under reduced pressure obtains 75% ethanol elution thing medicinal extract; C in () step (b), 75% ethanol elution medicinal extract purification on normal-phase silica gel is separated, obtain 5 components successively with the methylene chloride-methanol gradient elution that volume ratio is 80:1,50:1,30:1,15:1 and 1:1; D in () step (c), component 4 is separated further by purification on normal-phase silica gel, obtain 3 components successively with the methylene chloride-methanol gradient elution that volume ratio is 25:1,15:1 and 8:1; E in () step (d), component 2 reverse phase silica gel of octadecylsilane bonding is separated, with the methanol aqueous solution isocratic elution that concentration expressed in percentage by volume is 70%, collect 10 ~ 12 column volume elutriants, elutriant concentrating under reduced pressure obtains pure compound (I).
3. the preparation method of compound according to claim 2 (I), is characterized in that: described macroporous resin is AB-8 type macroporous adsorbent resin.
4. the preparation method of compound according to claim 2 (I), is characterized in that: it is 85% that described alcohol heat reflux extracts the alcohol concn adopted.
5. a pharmaceutical composition, is characterized in that: the compound according to claim 1 (I) wherein containing treatment significant quantity and pharmaceutically acceptable carrier.
6. compound according to claim 1 (I) application in the medicine preparing vascular protection.
7. the application of pharmaceutical composition according to claim 5 in the medicine preparing vascular protection.
CN201510672462.2A 2015-10-16 2015-10-16 New lignan compounds, and preparation method and medical application thereof Pending CN105152873A (en)

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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105237604A (en) * 2015-09-12 2016-01-13 徐建立 New limonin compound, preparation method and medical uses thereof
CN105523937A (en) * 2016-01-20 2016-04-27 温州统益生物医药科技有限公司 Diterpene compound with medical application and preparation method thereof
CN105622706A (en) * 2015-12-27 2016-06-01 温州统益生物医药科技有限公司 Triterpene compound for liver protection and preparation method thereof
CN112457284A (en) * 2019-09-09 2021-03-09 中国医学科学院药物研究所 Oligo-lignans compound, preparation method thereof, pharmaceutical composition thereof and application thereof

Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2003059340A1 (en) * 2002-01-17 2003-07-24 Hormos Medical Corporation Method for the preparation of lariciresinol, cyclolariciresinol and secoisolariciresinol
US20100129302A1 (en) * 2008-11-25 2010-05-27 Oy Granula Ab Ltd Antimicrobial composition with low cytotoxicity
CN105078945A (en) * 2015-09-05 2015-11-25 林天样 Application of Pedinophyllol B in preparation of vascular protective drug
CN105198896A (en) * 2015-09-16 2015-12-30 潘光贤 Clerodane diterpenoid compound and preparation method and medical application thereof
CN105218330A (en) * 2015-09-30 2016-01-06 宋晓梅 A kind of new Lignanoids compounds and preparation method thereof and medicinal use
CN105237604A (en) * 2015-09-12 2016-01-13 徐建立 New limonin compound, preparation method and medical uses thereof
CN105330521A (en) * 2015-10-22 2016-02-17 淄博夸克医药技术有限公司 New lignans compound, and preparation method and medical application thereof
CN105646164A (en) * 2015-12-18 2016-06-08 杭州市第人民医院 Neolignan compound for the treatment of gastric cancer

Patent Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2003059340A1 (en) * 2002-01-17 2003-07-24 Hormos Medical Corporation Method for the preparation of lariciresinol, cyclolariciresinol and secoisolariciresinol
US20100129302A1 (en) * 2008-11-25 2010-05-27 Oy Granula Ab Ltd Antimicrobial composition with low cytotoxicity
CN105078945A (en) * 2015-09-05 2015-11-25 林天样 Application of Pedinophyllol B in preparation of vascular protective drug
CN105237604A (en) * 2015-09-12 2016-01-13 徐建立 New limonin compound, preparation method and medical uses thereof
CN105198896A (en) * 2015-09-16 2015-12-30 潘光贤 Clerodane diterpenoid compound and preparation method and medical application thereof
CN105218330A (en) * 2015-09-30 2016-01-06 宋晓梅 A kind of new Lignanoids compounds and preparation method thereof and medicinal use
CN105330521A (en) * 2015-10-22 2016-02-17 淄博夸克医药技术有限公司 New lignans compound, and preparation method and medical application thereof
CN105646164A (en) * 2015-12-18 2016-06-08 杭州市第人民医院 Neolignan compound for the treatment of gastric cancer

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
张淑洁 等: ""厚朴化学成分及其现代药理研究进展"", 《中药材》 *
欧扬: ""川芎嗪二苯甲基哌嗪对氧化损伤血管内皮细胞的保护作用研究"", 《中国优秀硕士学位论文全文数据库医药卫生科技辑》 *

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105237604A (en) * 2015-09-12 2016-01-13 徐建立 New limonin compound, preparation method and medical uses thereof
CN105622706A (en) * 2015-12-27 2016-06-01 温州统益生物医药科技有限公司 Triterpene compound for liver protection and preparation method thereof
CN105523937A (en) * 2016-01-20 2016-04-27 温州统益生物医药科技有限公司 Diterpene compound with medical application and preparation method thereof
CN112457284A (en) * 2019-09-09 2021-03-09 中国医学科学院药物研究所 Oligo-lignans compound, preparation method thereof, pharmaceutical composition thereof and application thereof
CN112457284B (en) * 2019-09-09 2023-01-24 中国医学科学院药物研究所 Oligo-lignans compound, preparation method thereof, pharmaceutical composition thereof and application thereof

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Application publication date: 20151216