CN102731597A - Abelmoschus manihot extract and novel application of chemical components thereof - Google Patents
Abelmoschus manihot extract and novel application of chemical components thereof Download PDFInfo
- Publication number
- CN102731597A CN102731597A CN2012100187663A CN201210018766A CN102731597A CN 102731597 A CN102731597 A CN 102731597A CN 2012100187663 A CN2012100187663 A CN 2012100187663A CN 201210018766 A CN201210018766 A CN 201210018766A CN 102731597 A CN102731597 A CN 102731597A
- Authority
- CN
- China
- Prior art keywords
- extract
- flower
- compound
- sunset abelmoschus
- ethanol
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Abstract
The invention relates to an abelmoschus manihot extract and a novel application of chemical components thereof, concretely, relates to a compound of formula I or its pharmaceutically acceptable salt, solvate, ester and prodrug, and an extract containing the compound of formula I, a composition thereof and an application thereof. The compound or extract of the invention can be used as anti-oxidative damaging agent, and can be used for treating or preventing diseases or symptoms related to oxidative damage.
Description
Technical field
The present invention relates to the Chemistry for Chinese Traditional Medicine field; Relate to a kind of flavonoid compound that obtains that extracts from Flower of Sunset Abelmoschus; Also relate to this chromocor compound and comprise the new purposes of the Flos abelmoschi manihot extract of this chromocor compound, in particular as the purposes of anti-oxidative damage agent and be used to treat or the purposes of prevention and oxidative damage diseases associated or illness.
Background technology
Oxidative damage is to cause many chronic diseases, like cardiovascular diseases, cancer, hepatopathy, mellitus and the old and feeble important factor that takes place.Oxygen produces the active active oxygen of chemical property through the single electron reduction reaction in vivo, participates in the physiological process of body, and generation plays an important role with energy to the metabolism of organism.When active oxygen existed, superoxide-dismutase can be translated into oxygen, effectively prevented oxidative damage.But because overactivity oxygen exists; Or body self eliminate activity oxygen scarce capacity, cause the activity in vivo oxygen level to surpass arm's length standard, cause peroxo-such as protein, nucleic acid, lipid; And cause the damage of organism function, the most always cause the generation and the irreversible development of disease.
Although the transformation period of active oxygen is very short, because its chemical property is active, in short duration, can react with DNA, cause fracture of DNA nucleic acid chains and oxidative damage, it is crosslinked to form DNA.Can make protein denaturation with proteins react, influence proteinic normal physiological function.Cause the microbial film damage with lipid reactant, form the functional destruction of microbial film, thereby cause chronic diseases such as cardiovascular diseases, cancer, hepatopathy, mellitus and aging.Therefore, excessive active oxygen formation is a research content in modern life science field with eliminating in the control volume.
Many chronic diseases that oxidative damage causes for example cardiovascular diseases, cancer, hepatopathy, mellitus and aging etc. are well-known.For example; Liao Duanfang etc. (the anti-oxidative damage effect I of gypenoside, to the influence of radical damage cardiovascular function; Liao Duanfang, etc., Hengyang Medical College's journal; 1993 01 phases) disclose with guinea-pig heart papillary muscle and isolated rabbit aorta radical damage model, inquired into the provide protection of Herb Gynostemmae Pentaphylli total glycosides the radical damage cardiovascular function.The result finds that gypenoside (50mg/L) can reduce the myocardial contraction due to the Green Tea Extract, refractory phase shortens and autorhymicity increases; And can reduce the MDA of cardiac muscular tissue content, protection SOD activity and blood vessel endothelium discharge the ability of EDRF.
Chen Jihai etc. (relation of diabetes B microvascular complication and DNA oxidative damage, Chen Jihai etc., Jiangsu medicine, 2009 03 phases) have inquired into diabetes B (T2DM) merging microvascular complication patient's DNA oxidative damage.Its method is 87 routine T2DM patients to be divided into mellitus microvascular complication (A) group (n=56) do not have microvascular complication (B) group (n=31) with T2DM, and the normal examinees of alternative 65 examples are that normal control (C) is organized.Measure blood mononuclear cell DNA oxidative damage parameters-8 hydroxyl deoxy-guanine (8-OHdG) level with the ELISA method.A, B group blood 8-OHdG level all significantly raise than the C group as a result; The A group course of disease, 24h ALB, blood 8-OHdG level also significantly raise than the B group; Show that DNA oxidative damage that T2DM merges the microvascular complication patient more normally contrasts and not have the complication diabetic subject obvious, mellitus are merged the microvascular complication person, and to implement anti-oxidant treatment be rational.In addition, Yang Qianyong etc. (diabetic complication treatment New Policy: guard wire plastochondria oxidative damage, Yang Qianyong etc., medical science summary, 2007 24 phases) have summarized the oxidative damage that active oxygen (ROS) causes and in the pathogenesis of diabetic complication, have played an important role.It possibly be the common mechanism of diabetic complication morbidity that plastosome ROS increases.Therefore, reduce the oxidative damage that plastosome ROS generates and causes, for the control of diabetic complication provides New Policy.This New Policy is expected to realize through the medicine that designs Mitochondrially targeted property inhibitor and minimizing mitochondrial membrane potential.
The material that searching has an anti-oxidative damage ability is that those skilled in the art are expected to be used to treat or for example to prevent cardiovascular diseases, cancer, hepatopathy, mellitus and complication thereof and old and feeble chronic diseases.
Summary of the invention
The objective of the invention is to seek material to be used for treatment or to prevent for example cardiovascular diseases, cancer, hepatopathy, mellitus and complication thereof and old and feeble chronic diseases with anti-oxidative damage ability.The inventor finds to extract the monomeric compound Flower of Sunset Abelmoschus glycosides C that obtains and the extract that comprises this compound has tangible anti-oxidative damage ability from Flower of Sunset Abelmoschus.The present invention is based on this discovery and be accomplished.
For this reason, first aspect present invention provides with the following formula I compound:
Or its pharmacologically acceptable salts, solvate, ester, prodrug.
According to the compound of first aspect present invention, it is 3,3 ', 4 '; 5,5 ', 7,8-seven flavonols-8-O-β-the D-glucuronic acid is (promptly; 3,3 ', 4 ', 5; 5 ', 7,8-hepthydroxyl flavone-8-O-β-D-glucuronopyranoside) or its pharmacologically acceptable salts, solvate, ester, prodrug.In one embodiment, the pharmacologically acceptable salts of said compound is an alkali metal salt or alkaline earth salt, for example sylvite, sodium salt, magnesium salts etc.
Second aspect present invention provides a kind of Flos abelmoschi manihot extract, wherein comprises the described compound of first aspect present invention.
According to the extract of second aspect present invention, its ethanolic soln that to be Flower of Sunset Abelmoschus (be preferably greater than 60%, be preferably greater than 70%, 70-99% for example, 80-98% for example, 90-98% for example, for example about 95%) greater than 50% with concentration extracts and obtains.
According to the extract of second aspect present invention, it is that Flower of Sunset Abelmoschus (is preferably greater than 60%, is preferably greater than 70% greater than 50% with concentration; 70-99% for example; 80-98% for example, 90-98% for example, for example about 95%) ethanolic soln; Extract through heating (for example refluxing), extracting solution obtains after reclaiming solvent.
According to the extract of second aspect present invention, it is a Flower of Sunset Abelmoschus with concentration greater than 50% ethanolic soln, and through heating and extracting, extracting solution is after reclaiming solvent, and wash-out obtains on macroporous adsorbent resin (for example D101 resin) post.In one embodiment; It is to use following solvent elution successively that said wash-out is handled: (a) water or concentration are lower than 40% ethanol, (b) 40-85% ethanol (60-80% ethanol for example; For example about 70% ethanol), (c) 85-99% ethanol, the wherein extract of the present invention that after drying, obtains of (b) elutriated fraction.
According to the extract of second aspect present invention, wherein contain the described compound of first aspect present invention greater than 0.01% (for example 0.01-30%, for example 0.01-20%, for example 0.01-10%, for example 0.05-10%, for example 0.1-10%).
Third aspect present invention provides the method for preparing the said extract of second aspect present invention, and it may further comprise the steps:
(i) ethanolic soln that makes Flower of Sunset Abelmoschus (be preferably greater than 60%, be preferably greater than 70%, 70-99% for example, 80-98% for example, 90-98% for example, for example about 95%) greater than 50% with concentration, extracting solution desolventizes, and drying promptly gets extract; And, choose wantonly further
(ii) make step (i) products therefrom on macroporous adsorbent resin (for example D101 resin) post, use following solvent elution successively: (a) water or concentration are lower than 40% ethanol, (b) 40-85% ethanol (60-80% ethanol for example; For example about 70% ethanol), (c) 85-99% ethanol; Wherein (b) elutriated fraction obtains extract after drying.
According to the method for third aspect present invention, wherein the working concentration ethanolic soln that (is preferably greater than 60%, is preferably greater than 70%, 70-99% for example, 80-98% for example, 90-98% for example, for example about 95%) greater than 50% extracts Flower of Sunset Abelmoschus, obtains extract.
According to the method for third aspect present invention, wherein working concentration (is preferably greater than 60%, is preferably greater than 70% greater than 50%; 70-99% for example; 80-98% for example, 90-98% for example, for example about 95%) ethanolic soln; Extract Flower of Sunset Abelmoschus through heating (for example refluxing), extracting solution obtains extract after reclaiming solvent.
According to the method for third aspect present invention, wherein working concentration (is preferably greater than 60%, is preferably greater than 70% greater than 50%; For example 70-99%, for example 80-98%, for example 90-98%; For example about 95%) ethanolic soln; Extract Flower of Sunset Abelmoschus through heating (for example refluxing), extracting solution is after reclaiming solvent, and wash-out obtains on macroporous adsorbent resin (for example D101 resin) post.In one embodiment; It is to use following solvent elution successively that said wash-out is handled: (a) water or concentration are lower than 40% ethanol, (b) 40-85% ethanol (60-80% ethanol for example; For example about 70% ethanol), (c) 85-99% ethanol, the wherein extract of the present invention that after drying, obtains of (b) elutriated fraction.
According to the method for third aspect present invention, contain the described compound of first aspect present invention in its extract that obtains greater than 0.01% (for example 0.01-30%, for example 0.01-20%, for example 0.01-10%, for example 0.05-10%, for example 0.1-10%).
Fourth aspect present invention provides a kind of compsn, and it comprises said compound of first aspect present invention or the said extract of second aspect present invention, and optional physiology acceptable carrier.
According to the compsn of fourth aspect present invention, it is selected from: pharmaceutical composition, Halth-care composition, health food composition.
According to the compsn of fourth aspect present invention, it can be used as the anti-oxidative damage agent.
According to the compsn of fourth aspect present invention, it can be used for treatment or prevention and oxidative damage diseases associated or illness.
According to the compsn of fourth aspect present invention, it can be used for treating or prevents to include but not limited to following chronic disease: cardiovascular diseases, cancer, hepatopathy, mellitus and complication thereof, and old and feeble.
Fifth aspect present invention provides said compound of first aspect present invention or the purposes of the said extract of second aspect present invention in the product of preparation anti-oxidative damage agent.
Fifth aspect present invention also provides the said compound of first aspect present invention or the said extract of second aspect present invention to be used for treating or the purposes of the product of prevention and oxidative damage diseases associated or illness in preparation.
Fifth aspect present invention also provides the said compound of first aspect present invention or the said extract of second aspect present invention to be used to treat or to prevent to include but not limited to following chronic disease in preparation: cardiovascular diseases, cancer, hepatopathy, mellitus and complication thereof, and the purposes in the old and feeble product.
According to the purposes of fifth aspect present invention, wherein said product is medicine, healthcare products, protective foods.
Sixth aspect present invention provides the method for anti-oxidative damage in the experimenter who needs is arranged, and it comprises said compound of first aspect present invention or the said extract of second aspect present invention of using the anti-oxidative damage significant quantity to said experimenter.
Sixth aspect present invention also provides the method for in the experimenter who needs is arranged treatment or prevention and oxidative damage diseases associated or illness, and it comprises to said experimenter's administering therapeutic or prevents the said compound of first aspect present invention or the said extract of second aspect present invention of significant quantity.
Sixth aspect present invention also provides treats or prevents to include but not limited to following chronic disease in the experimenter who needs is arranged: cardiovascular diseases, cancer, hepatopathy, mellitus and complication thereof; And old and feeble method, it comprises said compound of first aspect present invention or the said extract of second aspect present invention to said experimenter's administering therapeutic or prevention significant quantity.
The characteristic that arbitrary embodiment had of the arbitrary aspect of the present invention is equally applicable to other arbitrary embodiment of this aspect; The characteristic that arbitrary aspect or its arbitrary embodiment of the present invention had is equally applicable to arbitrary embodiment of other arbitrary aspect or this other arbitrary aspect, as long as they can be not conflicting; Certainly at where applicable each other, necessary words can be done suitably to modify to individual features.
Do further to describe with characteristics to various aspects of the present invention below.
All documents that the present invention quoted from, their full content is incorporated this paper by reference into, and if the expressed implication of these documents and the present invention when inconsistent, be as the criterion with statement of the present invention.In addition; Various terms and phrase that the present invention uses have the general sense of well known to a person skilled in the art; Nonetheless; The present invention still hopes at this more detailed explanation and explanation to be done in these terms and phrase, and term of mentioning and phrase are as the criterion with the implication that the present invention was explained if any inconsistent with known implication.
In the present invention, arbitrary numerical range includes arbitrary subrange of this numerical range or the arbitrary concrete numerical value in this numerical range.For example when mentioning that extract of the present invention contains the described compound of first aspect present invention of 0.01-30%; Said 0.01-30% comprises arbitrary subclass or the arbitrary concrete numerical value in this numerical range; For example 0.01-30% includes but not limited to: 0.01-20%, 0.01-10%, 0.05-10%, 0.1-10%, or the like.
Because the present invention has verified that with monomeric compound it has a following effect to according to the invention: (1) anti-oxidative damage; (2) treatment or prevention and oxidative damage diseases associated or illness; (3) treatment or prevention include but not limited to following chronic disease: cardiovascular diseases, cancer, hepatopathy, mellitus and complication thereof and aging; Therefore The compounds of this invention can be any source; For example they can be chemosynthesis (comprising complete synthesis or semi-synthetic) independently of one another or obtain through from plant, extracting, and comprise that thus the extract expection of this compound has corresponding pharmacotoxicological effect equally.。
Of the present invention arbitrary aspect, when described compound or extract or compsn are used to bring into play the purpose that treats and/or prevents disease according to the invention or illness, in any case it is to use or preparation with the dosage that treats and/or prevents significant quantity.For example for pharmaceutical composition of the present invention; Effective dose wherein can be according to well known to a person skilled in the art the suitably processing of medical situation do; For example for the medicament of unitary dose; More after a little while, can obtain desired effects for activeconstituents in each unitary dose medicament through using a plurality of unitary dose medicaments; " significant quantity " and for example mentioned in the present invention, this significant quantity are that these compounds, extract or compsn offer said Mammals to be enough to the producing amount of expecting effect.In one embodiment of the invention; Reach the dosage of efficacious therapy and/or prevention; No matter what give is simplification compound, extract or pharmaceutical composition, this dosage is converted to the Flower of Sunset Abelmoschus medicinal material, is equivalent to medicinal material 0.001-50g/kg body weight/day; Preferred 0.01-25g/kg body weight/day, more preferably 0.1-10g/kg body weight/day.In one embodiment of the invention, for reaching the dosage of efficacious therapy and/or prevention, when with chemical monomer timing according to the invention; It can be the 0.001-100mg/kg body weight/day; Preferred 0.01-100mg/kg body weight/day, perhaps preferred 0.1-100mg/kg body weight/day, perhaps preferred 0.001-10mg/kg body weight/day; Perhaps preferred 0.001-1mg/kg body weight/day; Perhaps preferred 0.01-10mg/kg body weight/day, perhaps preferred 0.1-1mg/kg body weight/day, perhaps preferred 1-10mg/kg body weight/day.In addition, the clinician can confirm concrete using dosage according to many factors, and these factors are kind and the severity etc. of age, sex, general health situation, disease for example.In any case those skilled in the art openly can easily confirm the using dosage under the particular case according to the present invention.
The term that uses among this paper " extract " is meant the extract that is obtained through extraction by the vegetable drug Flower of Sunset Abelmoschus, wherein comprises formula I compound according to the invention at least, for example Flower of Sunset Abelmoschus glycosides C.Need to prove that can also comprise other material that exists originally in the Flower of Sunset Abelmoschus in this extract, these other materials are known in the art.
It is known to those skilled in the art that the content of measuring the formula I compound in medicinal material or the extract, these measuring methods are to obtain easily, for example can perhaps suitably do some adjustment with reference to the method for prior art bibliographical information.In the present invention, can adopt the HPLC method of following condition to measure the amount of the formula I compound in extract of the present invention or the compsn: chromatographic column: Hypersil-C18 chromatographic column (250mm * 4.6mm, 5 μ m); Moving phase is second eyeball/water=45/55; Flow velocity: 1.0mL/min; Detect wavelength: 254nm; Sample size: 10 μ L; 30 ℃ of column temperatures.In the present invention, during qualitative or quantitatively characterizing formula I compound of the present invention, in the time of perhaps need carrying out the preparation of monomeric compound,, all be to adopt this HPLC method when needs as explanation is not arranged in addition with the preparation HPLC method.
As described herein, term " significant quantity " is meant the dosage that can in the experimenter, realize treating, prevent, alleviate and/or alleviating disease according to the invention or illness.
As described herein, term " pharmaceutical composition " is meant to be used in the material of realizing treating, prevent, alleviate and/or alleviating disease according to the invention, illness, symptom among the experimenter.In special embodiment, should " pharmaceutical composition " can exchange use with " compsn ".
As described herein; Term " experimenter " can refer to that patient or other accept the present composition and extract to treat, to prevent, to alleviate and/or to alleviate the animal of disease according to the invention, illness, symptom; Particularly Mammals, for example people, dog, monkey, ox, horse etc.
As described herein, term " disease or symptom " is meant a kind of physical state of said experimenter, this physical state is relevant with disease according to the invention or symptom.
As described herein, " % ", as do not specialize, generally be meant the per-cent of w/w when being solid for total material, generally be meant the per-cent of weight/volume when being liquid for total material.Certainly, be liquid and solute when being liquid for total material, the per-cent that characterizes this liquid solute generally is meant the per-cent of volume.
" pharmaceutically acceptable carrier " that uses in the pharmaceutical composition of the present invention can be the carrier of any routine in the field of pharmaceutical preparations.The selection of specific support will be depended on administering mode or disease type and the state that is used to treat particular patient.The suitable drug preparation of compositions method that is used for the specific administration pattern is fully in pharmaceutical field technician's ken.For example, can be used as thinner, carrier, weighting agent, tackiness agent, wetting agent, disintegrating agent, absorption enhancer, tensio-active agent, absorption carrier and the lubricant etc. that pharmaceutically acceptable carrier comprises that pharmaceutical field is conventional.In case of necessity, can also in pharmaceutical composition, add flavouring agent, sanitas and sweeting agent etc.
Pharmaceutical composition of the present invention can be processed tablet, pulvis, granule, capsule, oral liquid, paste, creme, injectable emulsion various ways such as (sterile powder for injection pins).The medicine of above-mentioned various formulations all can be according to the ordinary method preparation of pharmaceutical field.
Embodiment
Can further describe the present invention through following embodiment, yet scope of the present invention is not limited to following embodiment.One of skill in the art can understand, and under the prerequisite that does not deviate from the spirit and scope of the present invention, can carry out various variations and modification to the present invention.The present invention carries out generality and/or concrete description to the material and the TP that are used in the test.Though for realizing that employed many materials of the object of the invention and working method are well known in the art, the present invention still does to describe in detail as far as possible at this.
Embodiment 1: the preparation of extract of the present invention and formula I compound
Step 1: the dried floral (2.8kg) of getting Flower of Sunset Abelmoschus; With the 95%EtOH solution of 10,8,8 times of amounts heating and refluxing extraction 3,2,2h successively, merging filtrate, decompression and solvent recovery; In 50 ℃ of following vacuum-dryings; Get 95% ethanol extraction (648g, 23.14%), this extract can be used as the reagent in the hereinafter Test Example.
Step 2: get above-mentioned steps 1 gained 95% ethanol extraction 500g; Handle (water → 70%EtOH → 95%EtOH), obtain water elution thing (220g, 10.2%), 70% ethanol elution thing (120g respectively through the D101 macroporous adsorbent resin; 5.55%) and 95% ethanol elution thing (22g, 1.02%).Wherein each eluate can be used as the reagent in the hereinafter Test Example, and particularly 70% ethanol elution thing can be used as the reagent in the hereinafter Test Example.
Step 3: above-mentioned steps 2 gained 70% ethanol elution thing (96g) is through silica gel column chromatography [CHCl
3-MeOH (100: 3 → 10: 1, v/v) → CHCl
3-MeOH-H
2O (7: 3: 1 → 6: 4: 1; V/v/v; Lower floor) → and MeOH] wash-out, obtain 9 elution fractions [9 elution fractions are followed successively by: Fr.1 (1.0g), Fr.2 (2.5g), Fr.3 (4.2g), Fr.4 (4.7g), Fr.5 (16.0g), Fr.6 (22.0g), Fr.7 (23.2g), Fr.8 (12.2g), Fr.9 (6.2g)] altogether.Wherein each eluate can be used as the reagent in the hereinafter Test Example.Particularly elution fraction Fr.9 can be used as the reagent in the hereinafter Test Example.
Step 4: above-mentioned steps 3 gained Fr.9 are through Sephadex LH-20 column chromatography [MeOH-H
2O (1: 1, v/v)], obtain 12 components [Fr.9-1～9-12].Wherein Fr.9-6 separates preparation [MeOH-H through preparative high performance liquid chromatography
2O (35: 65, v/v)+1%HAC], having obtained Flower of Sunset Abelmoschus glycosides C is 200mg.Use conventional salinization method, make the Flower of Sunset Abelmoschus glycosides C that obtains obtain its sodium salt with the sodium hydroxide salinization.
Embodiment 2: the structure of Flower of Sunset Abelmoschus glycosides C is identified and physico-chemical property
The Flower of Sunset Abelmoschus glycosides C that embodiment 1 step 4 obtains: amorphous yellow powder.Be a yellow blackening under the 254nm uv lamp, the colour developing of 10% sulfuric acid ethanol liquid is pale brown look, and supposition possibly be flavonoid compound.Its specific rotatory power is negative value ([α]
D 25:+49.0 °, MeOH).High resolution Q-TOF-ESI-MS provides its quasi-molecular ion peak m/z 509.0554 [M-H]
-, confirm that its molecular formula is C
21H
18O
15There is hydroxyl (3342cm in the information display structure that ir spectra provided
-1), unsaturated ketone group (1652cm
-1), aromatic nucleus (1607,1511,1418cm
-1).UV spectrum further point out the existing of flavonol skeleton (305,255nm).
1H-NMR (400MHz, DMSO-d
6) low place δ 12.44 in the spectrum (and 1H, br.s) in the prompting structure 5 have 1 phenolic hydroxyl group; (2H is that the sharp-pointed unimodal B of a supposition ring is 1,3,4 s), 5-or 1,2,4,6 symmetries, four replacement spin coupled systems by fragrance district δ 7.44; (1H is one sharply unimodal s) to δ 6.25, infers that it possibly encircle 6 proton signals for A.In addition, high field region sugar end group signal δ 4.71 (1H, d, J=7.6Hz) and sugar go up other proton signal δ 3.58 (1H, d, J=9.2Hz), 3.44 (1H, m), 3.40 (1H, m), 3.32 (1H, m) with
13C-NMR (100MHz, DMSO-d
6) in one group of associated carbon signal (δ
C: 71.6,73.9,75.5,75.9,106.0,171.3) be connected with 1 β-D-glucal acid fragment in the supposition structure.In addition, in the HMBC spectrum, can observe H-6 [δ
H6.25 (1H, s)] and C-10 (δ
C102.7), C-8 (δ
C125.0), C-5 (δ
C156.3) and C-7 (δ
C156.2), H-2 ', 6 ' [δ
H7.44 (2H, s)] and C-1 ' (δ
C120.4), C-4 ' (δ
C135.9), C-3 ', 5 ' (δ
C145.5) and C-2 (δ
C147.9), H-1 " [δ
H4.71 (1H, d, J=7.6Hz)] and C-8 (δ
C125.0), 5-OH proton [δ
H11.92 (1H, s)] and C-6 (δ
C98.2), C-10 (δ
C102.7), C-5 (δ
C156.3) between long-range relevant.This compound is through 5%H
2SO
4The aqueous solution-1, (the sugared product after the hydrolysis is the BSTFA derivatize after CMEH handles, and through GC its tetrahydrothiazole derivates is analyzed again for 1:1, v/v) acid hydrolysis, has confirmed to connect in the structure sugar and has been the D-glucuronic acid for the 4-dioxane.In sum, the structure of confirming Flower of Sunset Abelmoschus glycosides C is 3,3 ', 4 ', 5,5 ', 7,8-seven flavonols-8-O-β-D-glucuronic acid (3,3 ', 4 ', 5,5 ', 7,8-hepthydroxyl flavone-8-O-β-D-glucuronopyranoside).Flower of Sunset Abelmoschus glycosides C wherein
1H-
1H COSY and HMBC dependency structure are illustrated as follows:
The physico-chemical property of Flower of Sunset Abelmoschus glycosides C:
Yellow powder.[α]
D 25+ 49.0 ° (concentration 0.31, MeOH).
IRλ
max(KBr)cm
-1：3342、2911、1721、1652、1607、1568、1511、1418、1329、1252、1202、1174、1072、1023、979、839、794。
UVν
max(MeOH)nm(logε)：305(3.79)、255(4.14)。
HR-ESI-TOF-MS: negative ion mode m/z 509.0554 [M-H]
-(calculated value, C
21H
17O
15, 509.0552), 545.0342 [M+Cl]
+(calculated value, C
21H
18O
15Cl:545.0340).
1H-NMR (DMSO-d
6, 400MHz): δ 3.44 (1H, m, overlapping, H-2 "), 3.58 (1H, d; J=9.2Hz, H-5 "), 3.33 (and 1H, m, overlapping, H-3 "), 3.38 (1H, m; overlapping, H-4 "), 4.71 (1H, d, J=7.6Hz, H-1 "), 6.25 (1H, s; H-6), 7.44 (2H, s, H-2 ', 6 '), 12.44 (1H, br.s, 5-OH).
13C-NMR(DMSO-d
6，100MHz)：δ
C：147.9(C-2)、135.5(C-3)、175.6(C-4)、156.3(C-5)、98.2(C-6)、156.2(C-7)、125.0(C-8)、148.2(C-9)、102.7(C-10)、120.4(C-1′)、107.7(C-2′)、145.5(C-3′)、135.9(C-4′)、145.5(C-5′)、107.7(C-6′)、106.0(C-1″)、73.9(C-2″)、75.5(C-3″)、71.6(C-4″)、75.9(C-5″)、171.3(C-6″)。
Test Example 1: Flower of Sunset Abelmoschus 95% ethanol extraction and chemical ingredients Study on antioxidation
The HepG2 cell is a human liver cancer cell; Because of it has preserved a lot of normal liver cells' metabolic characteristic; Comprise synthetic and the secretion plasma proteins, and be widely used as studying the external model of mechanism of drug action, toxicity/cytoprotective, genetoxic/livers such as anti-genetoxic protection experiment.(tert-butylhydroperoxide can be a living radical by metabolism such as Cytochrome P450s t-BHP) to tertbutyl peroxide, and the latter can further produce active oxygen radical, thereby causes peroxidatic reaction of lipid, causes cell injury.
TP: with cell inoculation in 96 orifice plates, 1 * 10
4Cells/well, 24h is cultivated in 5 every group multiple holes, adds embodiment 1 step 1 gained Flower of Sunset Abelmoschus 95% ethanol extraction and Flower of Sunset Abelmoschus glycosides C.Other establishes normal control group (promptly also handling the not test group of administration with t-BHP), model group (promptly handle with t-BHP but not the test group of administration).Behind the application of sample 20h, except that the normal control group, all the other each group add t-BHP to concentration be 0.5mmol/L, damage after 7 hours, mtt assay mensuration is respectively organized the survival rate of cell.
With cell inoculation in 96 orifice plates, 3 * 10
4Cells/well, 24h is cultivated in 5 every group multiple holes, adds embodiment 1 step 1 gained Flower of Sunset Abelmoschus 95% ethanol extraction and Flower of Sunset Abelmoschus glycosides C.Other establishes normal control group, model group.Behind the application of sample 20h, except that the normal control group, all the other each group add t-BHP to concentration be 0.5mmol/L, damage after 5 hours, carefully sop up cell conditioned medium liquid, add fresh medium and cultivate 12h, absorb supernatant, the ultrasonication cell is measured the interior GSH content of cell.
With cell inoculation in 24 orifice plates, 2.5 * 10
5Cells/well, 24h is cultivated in 4 every group multiple holes, adds embodiment 1 step 1 gained Flower of Sunset Abelmoschus 95% ethanol extraction and Flower of Sunset Abelmoschus glycosides C.Other establishes normal control group, model group.Behind the application of sample 20h, except that the normal control group, all the other each group add t-BHP to concentration be 0.5mmol/L; Damage after 5 hours, carefully sop up cell conditioned medium liquid, add fresh medium and cultivate 12h; Get supernatant 200 μ l and measure MDA content; 400 μ lPBS ultrasonication cells are got 200 μ l and are measured the CAT vigor, and 50 μ l measure the SOD vigor.
Testing data is represented with mean ± standard deviation
; Adopt the SPSS11.5 software package to carry out statistical procedures, significance level is a standard with 0.05.Test-results is seen table 1.
Table 1:
* *P＜0.001,
*P＜0.01,
*P＜0.05, the model group of handling with t-BHP compares.
T-BHP can make the HepG2 cell viability descend, and SOD, CAT vigor and GSH content obviously descend in the cell, and MDA content rises in the cell.Flower of Sunset Abelmoschus 95% alcohol extract is (through measuring; Contain 0.05% Flower of Sunset Abelmoschus glycosides C in this reagent), Flower of Sunset Abelmoschus glycosides C can suppress the decline of cell viability due to the t-BHP; The reduction of GSH due to can inhibition t-BHP in various degree, suppress MDA in the impaired HepG2 cell conditioned medium liquid rising, improve GSH content and CAT, SOD vigor in the cell, have very strong anti-oxidant activity.
In addition; The contriver with the other test of above-mentioned experimental example 1 same procedure in; Used embodiment 1 step 2 gained 70% ethanol elution thing (, containing 0.21% Flower of Sunset Abelmoschus glycosides C in this reagent) respectively through measuring, the two tests as reagent step 3 gained elution fraction Fr.9 (through measuring, containing 3.2% Flower of Sunset Abelmoschus glycosides C in this reagent); The dosage of two kinds of reagents is in contained Flower of Sunset Abelmoschus glycosides C, the Flower of Sunset Abelmoschus glycosides C when dosage such as has with 95% ethanol extraction test group in the table 1.The model group that the result of 4 test parameters of two kinds of reagents of result (GSH, MDA, CAT in cell survival rate, the cell) and t-BHP handle relatively is
*P＜0.01, and the model group that SOD value and t-BHP handle relatively is
*P＜0.05.
Claims (10)
1. formula I compound:
Or its pharmacologically acceptable salts, solvate, ester, prodrug.
2. Flos abelmoschi manihot extract wherein comprises the described compound of claim 1.
3. the extract of claim 2, to be Flower of Sunset Abelmoschus with concentration extract greater than 50% ethanolic soln obtains for it; Or choose wantonly further, extracting solution obtains after reclaiming solvent.
4. the extract of claim 2, it is a Flower of Sunset Abelmoschus with concentration greater than 50% ethanolic soln, and through heating and extracting, extracting solution is after reclaiming solvent, and wash-out obtains on macroporous adsorptive resins.
5. each extract of claim 2-4 wherein contains greater than the described compound of 0.01% claim 1.
6. prepare the method for each said extract of claim 2-5, it may further comprise the steps:
(i) make Flower of Sunset Abelmoschus with concentration greater than 50% ethanolic soln, extracting solution desolventizes, drying promptly gets extract; And, choose wantonly further
(ii) make step (i) products therefrom on macroporous adsorptive resins, use following solvent elution successively: (a) water or concentration are lower than 40% ethanol, (b) 40-85% ethanol (60-80% ethanol for example; For example about 70% ethanol), (c) 85-99% ethanol; Wherein (b) elutriated fraction obtains extract after drying.
7. the method for claim 6 is characterized in that:
Working concentration extracts Flower of Sunset Abelmoschus greater than 50% ethanolic soln, obtains extract;
Working concentration is greater than 50% ethanolic soln, and through the heating and extracting Flower of Sunset Abelmoschus, extracting solution obtains extract after reclaiming solvent; Or
Working concentration is greater than 50% ethanolic soln, and through the heating and extracting Flower of Sunset Abelmoschus, extracting solution is after reclaiming solvent, and wash-out obtains on macroporous adsorptive resins.
8. compsn, it comprises said compound of claim 1 or the said extract of claim 2-5, and optional physiology acceptable carrier.
9. the compsn of claim 8, it is as the anti-oxidative damage agent; Perhaps be used for treatment or prevention and oxidative damage diseases associated or illness; Perhaps be used to treat or prevent to include but not limited to following chronic disease: cardiovascular diseases, cancer, hepatopathy, mellitus and complication thereof, and old and feeble.
10. the purposes of said compound of claim 1 or the said extract of claim 2-5 is used to prepare the product of anti-oxidative damage agent; Perhaps be used to prepare and be used to treat or the product of prevention and oxidative damage diseases associated or illness; Perhaps be used to prepare and be used to treat or prevent to include but not limited to following chronic disease: cardiovascular diseases, cancer, hepatopathy, mellitus and complication thereof, and old and feeble product.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210018766.3A CN102731597B (en) | 2012-01-20 | 2012-01-20 | Abelmoschus manihot extract and novel application of chemical components thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210018766.3A CN102731597B (en) | 2012-01-20 | 2012-01-20 | Abelmoschus manihot extract and novel application of chemical components thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102731597A true CN102731597A (en) | 2012-10-17 |
| CN102731597B CN102731597B (en) | 2014-10-29 |
Family
ID=46987970
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201210018766.3A Active CN102731597B (en) | 2012-01-20 | 2012-01-20 | Abelmoschus manihot extract and novel application of chemical components thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102731597B (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103911041A (en) * | 2014-03-31 | 2014-07-09 | 苏州科斯伍德油墨股份有限公司 | Invariant color oil-based ink containing abelmoschus manihot extract and preparation method of ink |
| CN103976466A (en) * | 2014-05-20 | 2014-08-13 | 河南中烟工业有限责任公司 | Abelmoschus manilhot seed extractive and application in cigarette |
| CN104644699A (en) * | 2013-11-18 | 2015-05-27 | 童玉新 | Sunset abelmoschus flower extract used for treatment of skin warts, and preparation and application thereof |
| CN108379330A (en) * | 2018-05-12 | 2018-08-10 | 济南昊雨青田医药技术有限公司 | A kind of extraction purification technology of antivirus effective position |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1994337A (en) * | 2006-11-11 | 2007-07-11 | 周亚球 | Total flavone extract of maniod eibish, its preparation and application |
-
2012
- 2012-01-20 CN CN201210018766.3A patent/CN102731597B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1994337A (en) * | 2006-11-11 | 2007-07-11 | 周亚球 | Total flavone extract of maniod eibish, its preparation and application |
Non-Patent Citations (2)
| Title |
|---|
| XIANYIN LAI,ET AL.,: "SPE-HPLC method for the determination of four flavonols in rat plasma and urine after oral administration of Abelmoschus manihot extract", 《JOURNAL OF CHROMATOGRAPHY B》 * |
| 刘爽等: "黄蜀葵化学成分及药理活性研究进展", 《中国现代中药》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104644699A (en) * | 2013-11-18 | 2015-05-27 | 童玉新 | Sunset abelmoschus flower extract used for treatment of skin warts, and preparation and application thereof |
| CN103911041A (en) * | 2014-03-31 | 2014-07-09 | 苏州科斯伍德油墨股份有限公司 | Invariant color oil-based ink containing abelmoschus manihot extract and preparation method of ink |
| CN103911041B (en) * | 2014-03-31 | 2016-02-10 | 苏州科斯伍德油墨股份有限公司 | Nondiscoloration oil-based ink containing Flos abelmoschi manihot extract and preparation method thereof |
| CN103976466A (en) * | 2014-05-20 | 2014-08-13 | 河南中烟工业有限责任公司 | Abelmoschus manilhot seed extractive and application in cigarette |
| CN108379330A (en) * | 2018-05-12 | 2018-08-10 | 济南昊雨青田医药技术有限公司 | A kind of extraction purification technology of antivirus effective position |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102731597B (en) | 2014-10-29 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP4669920B2 (en) | Functional material that suppresses blood glucose rise and blood pressure rise | |
| AU2014314827B2 (en) | New salvianolic acid compound T, preparation method therefor, and use thereof | |
| RU2559784C2 (en) | Composition for mitigating fatigue, formulation and using them | |
| EP3560506A1 (en) | Pharmaceutical composition comprising indigo pulverata levis extract or fraction thereof as effective ingredient for preventing or treating inflammatory bowel disease | |
| TW201622698A (en) | Compositions including a chrysanthemum extract and methods of using the same in skin whitening and skin lightening applications | |
| Perera et al. | Perspectives on geraniin, a multifunctional natural bioactive compound | |
| CN102731597B (en) | Abelmoschus manihot extract and novel application of chemical components thereof | |
| Wang et al. | Diversity of potentially exploitable pharmacological activities of the highly prized edible medicinal fungus Antrodia camphorata | |
| WO2016043517A1 (en) | Pharmaceutical composition for treating and preventing degenerative neurological disorders, containing, as active ingredient, mixture extract of moutan root bark, angelica dahurica root and bupleurum root or fraction thereof | |
| CN107496573B (en) | Application of houttuynia cordata extract in preparation of pharmaceutical preparation for preventing myocardial ischemia reperfusion injury | |
| CN106822166B (en) | A kind of drug for preventing and treating diabetes and hyperlipidemia and its application in pharmacy | |
| KR100979459B1 (en) | Tetracera scandens extracts and 4H-chromen-4-one derivatives isolated therefrom increasing glucose uptake in differentiated L6 muscle cells | |
| KR101160088B1 (en) | Herbal extracts composition for the prevention of alcoholic fatty liver, hyperlipidemia and hangover | |
| KR20210047594A (en) | Compositions for reinforcing skin barrier and improving atopic dermatitis using hydrangenol or phyllodulcin as an active ingredient | |
| KR20200063772A (en) | Composition for anti-inflammation comprising extract of mealworm containing ginsenoside Re | |
| JP2007031302A (en) | Adiponectin production accelerator and metabolic syndrome preventive | |
| CN104367612B (en) | A kind of application of dog ant grass extract | |
| KR101402058B1 (en) | Composition comprising extract of fermented Curcuma longa by natural fermented soybean and curcuminoid derivatives isolated therefrom for treating or preventing liver disease | |
| WO2011095095A1 (en) | Medicinal composition comprising alcohol-soluble and water-insoluble licorice extract, pharmaceutical preparation, pharmaceutical application, therapeutic method, and preparative method thereof | |
| CN102526387B (en) | Medicinal composition for treating early-stage diabetic foot and preparation method thereof | |
| JP2010168399A (en) | Pharmaceutical for controlling elevation of blood sugar | |
| CN1899410A (en) | Medicine for treating cardiovascular disease and its preparing method and quality control method | |
| KR100767051B1 (en) | Composition comprising obovatol and/or obovatal, Pharmaceutically acceptable salts thereof, or Their derivatives as active ingredient for Curing and Preventing fatness, and Purification method of the active ingredients | |
| KR20180111280A (en) | Composition for preventing and treating inflammatory bowel disease comprising sagassum serratifolium extract | |
| CN108904481A (en) | O-hydroxylate chalcone analog is preparing the application in anti-oxidation medicine |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |