CN1241559C - Chinese traditional medicine hypoglycemic agent and preparing process thereof - Google Patents
Chinese traditional medicine hypoglycemic agent and preparing process thereof Download PDFInfo
- Publication number
- CN1241559C CN1241559C CN 02138362 CN02138362A CN1241559C CN 1241559 C CN1241559 C CN 1241559C CN 02138362 CN02138362 CN 02138362 CN 02138362 A CN02138362 A CN 02138362A CN 1241559 C CN1241559 C CN 1241559C
- Authority
- CN
- China
- Prior art keywords
- ethanol
- chinese medicine
- ethyl acetate
- extract
- concentrate
- 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.)
- Expired - Fee Related
Links
Abstract
The present invention extracts and separates effective parts and effective components which have the function of lowering blood sugar from winged euonymus twig which is traditional Chinese medicine. An ethyl acetate extract of the winged euonymus twig is separated by macroporous resin, and 50% of ethanol elution part, terpene and phenolic acid as well as EA-2 and EA-3 are separated. The present invention has the function of lowering blood sugar of a diabetic mouse and has little and low side effect. If a patient takes the present invention for a long time, considerable safety can be still obtained. Malondialdehyde and superoxide dismutase are influenced. The present invention has the function of treating both manifestation and root cause of a disease. Occurrence and development of diabetic complication can be lightened or inhibited.
Description
Technical field
The present invention relates to utilize Chinese medicine to prepare blood sugar lowering and preparation method thereof, relate in particular to and adopt modern extraction, isolation technics, from the Chinese medicine Ramulus Euonymi, obtain to have the effective site and the effective ingredient of hypoglycemic activity, the preparation blood sugar lowering.
Background technology
Diabetes are a kind of commonly encountered diseases, are difficult to cure, and needs of patients is taken medicine for a long time, and diabetic complication often causes serious consequence.The diabetes Chinese medical discrimination belongs to " quenching one's thirst " sick category.Study carefully its pathogenesis and mainly be YIN fluid deficiency, scorching inclined to one side victory, and be this with the deficiency of YIN scorchingly is mark.Production of dryness-heat in the interior, the dried liquid in Tianjin is withered, and QI and blood is not smooth, is the stasis of blood for a long time.Clinical medicine commonly used has metformin hydrochloride and insoral (phenformin) at present, is biguanide antidiabetic medicament.The insoral major side effects is a gastrointestinal reaction, anorexia, bitter taste are arranged, feels sick, symptoms such as vomiting, abdominal distention and diarrhoea, because of through liver metabolism, easily brings out lactic acidosis.Though the probability that metformin brings out lactic acidosis still has this side effect than the remarkable minimizing of insoral, and major part occurs in this medicine had the contraindication patient, as the renal failure patient.Therefore, need research and development good effect, few side effects and the modern preparation of blood sugar lowering Chinese medicine little, treating both the principal and secondary aspects of a disease.
Ramulus Euonymi is aliform appurtenance or its twig of Celastraceae plant winged euonymus Euonymus alatus (Thunb.) Sieb., and sour in the mouth is sweet puckery, and cold nature can restore menstrual flow and invigorate blood circulation.The report of the domestic and international relevant rarely seen sterol of its The Chemical Constituents, flavone compound, referring to: Chen Ke etc., the steroidal composition [J] in the Celastraceae plant Ramulus Euonymi. Chinese herbal medicine (Chin Trait Herb Drugs), 1983,14 (9): 385-388; Chen Ke etc., the flavane composition [J] in the Ramulus Euonymi. Chinese herbal medicine (Chin Trait Herb Drugs), 1986,17 (3): 97-100; Susumu Kitanaka etal.Cytotoxic cardenolides from woods of Euonymus alata[J] .Chem Pharm Bull, 1996,44 (3): 615-617.
Summary of the invention
The technical problem to be solved in the present invention is from Ramulus Euonymi, research and development good effect, few side effects and little hypoglycemic effective site and the modern preparation of Chinese medicine of effective ingredient.Side effect significantly reduces and reduces, and still has suitable safety even make the patient take for a long time; Add and have the treating both the principal and secondary aspects of a disease function, can alleviate or suppress the generation and the development of diabetic complication.The technical problem to be solved in the present invention also comprises extraction, the separation method of the above-mentioned blood sugar lowering Chinese medicine composition of research preparation, and this method is answered favorable reproducibility, can be used in suitability for industrialized production.
For solving the problems of the technologies described above, technical scheme of the present invention is as follows.
A kind of hypoglycemic Chinese medicine composition is characterized in that active component makes with the following method:
1) get the Ramulus Euonymi medical material, water and/or ethanol extraction concentrate extracting solution, obtain concentrate;
2) get the concentrate of water in the step 1) and/or ethanol extract, use ethyl acetate extraction, concentrated extract obtains acetic acid ethyl ester extract, is the active component of described Chinese medicine composition; Perhaps, get the concentrate of water in the step 1) and/or ethanol extract, carry out chromatography with 15-60 order polyamide, use the aquiferous ethanol gradient elution, collect its ethanol elution position of 50%, after concentrating, obtain the mixture of terpenoid and phenolic acid compound, be the active component of described Chinese medicine composition.Described acetic acid ethyl ester extract further carries out chromatography with 15-60 order polyamide, uses the aquiferous ethanol gradient elution, collects its ethanol elution position of 50%.The application of the active component acetic acid ethyl ester extract of Chinese medicine composition in preparation blood sugar lowering medicine.
A kind of hypoglycemic Chinese medicine composition is characterized in that active component makes with the following method:
1) gets the Ramulus Euonymi medical material, use ethyl acetate extraction, extracting solution is concentrated, obtain concentrate;
2) get the concentrate of acetic acid ethyl acetate extract in the step 1), carry out chromatography, use the aquiferous ethanol gradient elution with 15-60 order polyamide, collect its ethanol elution position of 50%, after concentrating, obtain the mixture of terpenoid and phenolic acid compound, be the active component of described Chinese medicine composition.
The application in preparation blood sugar lowering medicine of the active component terpenoid of aforementioned Chinese medicine composition and the mixture of phenolic acid compound.Described Chinese medicine composition contains pharmaceutically acceptable carrier.That is: described kinds of ingredients or chemical constituent can with pharmaceutically be fit to carrier and mix, make various preparations.
A kind of hypoglycemic Chinese medicine composition mainly is made up of the ethyl acetate extract or the extract of Ramulus Euonymi.Described Chinese medicine composition, the ethyl acetate extract of Ramulus Euonymi or extract mainly are made up of compound of phenolic acid and/or terpenoid.Compound of phenolic acid is mainly P-hydroxybenzoic acid, 3,4-resorcylic acid, 3-methoxyl group-4-hydroxy benzoic acid and/or 3.5-dimethoxy-4 '-hydroxy benzoic acid.
A kind of hypoglycemic Chinese medicine composition preparation method, it is characterized in that: 1) get the Ramulus Euonymi medical material, water, ethanol and/or ethyl acetate extraction get water extraction concentrate, ethanol extraction concentrate or ethyl acetate extraction concentrate after concentrating, or its corresponding concentrate; 2) water extraction concentrate or ethanol extraction concentrate, or the concentrate of main water or ethanol extraction are used ethyl acetate extraction, get the acetic acid ethyl ester extract that mainly contains terpenoid and phenolic acids composition; Acetic acid ethyl ester extract carries out chromatography with 15-60 order polyamide, uses the aquiferous ethanol gradient elution, and its ethanol elution position of 50% obtains the mixture of terpenoid and phenolic acid compound; Perhaps, water extraction concentrate or ethanol extraction concentrate, or the concentrate of main water or ethanol extraction, carry out chromatography with 15-60 order polyamide, use the aquiferous ethanol gradient elution, its ethanol elution position of 50%, the reuse ethyl acetate extraction obtains the mixture of terpenoid and phenolic acid compound; Perhaps, ethyl acetate extraction concentrate or mainly use the concentrate of ethyl acetate extraction carries out chromatography with 15-60 order polyamide, uses the aquiferous ethanol gradient elution, and its ethanol elution position of 50% obtains the mixture of terpenoid and phenolic acid compound.
Preceding method is characterized in that: main water or ethanol extraction or mainly use ethyl acetate extraction, be meant in its extracting solution that water or ethanol or ethyl acetate content are 30～99%, with 51～99% for well; Mainly use ethyl acetate extraction, be meant that ethyl acetate content is 30～99% in its extracting solution, with 51～99% for well.
Said method is characterised in that: 50% ethanol elution position of polyamide chromatography, through the SephadexLH-20 chromatography, use chloroform again: and 1: 1 eluting of methanol, flow out terpenoid earlier, the phenolic acids component is flowed out in the back.
Described method is characterised in that: the phenolic acids component is carried out silica gel column chromatography, use the chloroform-methanol gradient elution, again through Sephadex LH-20 column chromatography for separation, use chloroform: 2: 1 gradient elutions of methanol, obtain chemical compound P-hydroxybenzoic acid, 3 respectively, 4-resorcylic acid, 3-methoxyl group-4-hydroxy benzoic acid and 3.5-dimethoxy-4 '-hydroxy benzoic acid.
The application in preparation blood sugar lowering medicine of the ethyl acetate extract of Ramulus Euonymi, acetic acid ethyl ester extract, terpenoid and/or phenolic acids component.
P-hydroxybenzoic acid, 3, the application in preparation blood sugar lowering medicine of 4-resorcylic acid, 3-methoxyl group-4-hydroxy benzoic acid and/or 3.5-dimethoxy-4 '-hydroxy benzoic acid.
The present invention follows the trail of the effective site and the active component of Ramulus Euonymi blood sugar lowering based on its activity.Experiment is set about from different extract parts, select normal and chemical diabetes mouse model for use, on integral level, carry out antidiabetic drug and imitate relatively, the effective site of preliminary definite Ramulus Euonymi blood sugar lowering is the ethyl acetate extraction position of Ramulus Euonymi decocting liquid, carry out polyamide column chromatography again, get its ethanol elution position of 50% and have good efficacy.Four liposoluble ingredients that obtain having curative effect are separated at ethanol elution position from 50%, their structure is defined as P-hydroxybenzoic acid (being called for short EA-1) through chemical method and Spectrum Analysis, 3,4-resorcylic acid (being called for short EA-2), 3-methoxyl group-4-hydroxy benzoic acid (being called for short EA-3), 3.5-dimethoxy-4 '-hydroxy benzoic acid (being called for short EA-4).Four chemical compounds are to separate from this plant first and obtain.
Diabetes are that islet cells is impaired, hypoinsulinism and a kind of endocrinopathy of the metabolism disorder that causes.It is generally acknowledged that hypoglycemic mechanism of action relates generally to two aspects: the one, promote secretion of insulin and the release or the secretion of glucagon suppression; The 2nd, influence metabolic some link behind the Insulin receptor INSR, mainly be the glyconeogenesis that suppresses liver, its purpose all is to reduce and blood sugar control.Experimental result shows that Ramulus Euonymi does not have obvious influence to healthy mice blood glucose, points out it significantly not stimulate insulin release action or ILA; The effective site of extraction separation and effective ingredient have tangible reduction effect to the blood sugar increasing due to the alloxan in the Ramulus Euonymi under the low dosage condition, and this shows the function of the β cell that it all has repairing damage or reduces the antagonism of body to insulin.
The diabetes Chinese medical discrimination belongs to " quenching one's thirst " sick category.Study carefully its pathogenesis and mainly be YIN fluid deficiency, scorching inclined to one side victory, and be this with the deficiency of YIN scorchingly is mark.Production of dryness-heat in the interior, the dried liquid in Tianjin is withered, and QI and blood is not smooth, is the stasis of blood for a long time.So the treatment diabetes are worked as based on replenishing YIN and removing heat, blood circulation promoting and blood stasis dispelling.Ramulus Euonymi nature and flavor bitter cold, thus can consolidating YIN heat clearing away, the merit of blood circulation promoting and blood stasis dispelling is arranged again.The effective site of extraction separation and effective ingredient in the Ramulus Euonymi, it is controlled to quench one's thirst and can take into account specimen, and the diabetes pathogenesis hits.
The present invention has studied aforementioned 50% ethanol elution position, terpenoid, phenolic acids and EA-2, the EA-3 of the Ramulus Euonymi influence to blood glucose in diabetic mice, malonaldehyde (MDA), superoxide dismutase (SOD).
Kunming mouse, body weight 22-25 gram, male and female half and half are provided by this school Animal House.(body weight 22～25g) is raised fasting 16 hours (drinking-water is not limit) after 3 days to healthy mice, and (with water recently distilled preparation, concentration 1%, dosage: 200mg/kg), the normal raising after 72 hours, surveyed fasting blood sugar (FSG) to the lumbar injection alloxan.The mice of choosing FSG＞180mg/kg is a diabetic mice.
Administration and mensuration: 12 of first picked at random healthy mices are as normal group before the modeling.Choose 84 of diabetic mices after other mice modelings, 12 every group, be divided into 7 groups at random by fasting blood sugar.Each treatment group I (50% ethanol) 3.2mg/Kg; II (terpenoid) 3.00mg/Kg; III (phenolic acid) 3.00mg/Kg; IV (EA3) 1.5mg/Kg; V (EA2) 1.5mg/Kg, normal group and model group are given the CMC-Na of equivalent 0.5%, and the positive drug group is given and insoral (110mg/kg).Administering mode is pressed 0.2ml/10g and is irritated stomach.Successive administration 7 days, last administration fasting in preceding 2 hours, two hours after administration eye socket rear vein beard is got blood, presses the test kit explanation and surveys blood glucose, MDA, SOD.
Influence to blood glucose: table 1 data show that Ramulus Euonymi ethyl acetate extract polyamide column has tangible blood sugar reducing function with 50% ethanol elution position and EA-2.In the experiment, contrast medicine insoral is to calculate by the labelled amount of tablet, other test dose then be by extract rate conversion (with crude drug 4g/kg is reference, the yield of extract * 4g/kg).
Table 1, each position of Ramulus Euonymi to tetrahydropyrimidine induce blood glucose in diabetic mice effect (n=12, X ± S)
Group dosage blood glucose value (mg/dl) blood glucose decline percentage rate
(mg/kg) after the preceding administration of administration (%)
Normal group/138.70 ± 22.10 119.95 ± 25.91 13.5
Model group/542.5 ± 202.19 531.18 ± 190.64 2.1
Insoral 110 544.6 ± 199.53 327.4 ± 190.42*## 39.9
50% ethanol, 3.20 545.1 ± 198.43 371.82 ± 153.81*# 31.8
Terpenoid 3.00 542.2 ± 186.21 409.37 ± 167.82 24.5
Phenolic acid 3.00 547.8 ± 185.88 542.88 ± 152.32 0.9
EA-3 1.50 552.7±199.72 421.65±242.80 23.7
EA-2 1.50 572.7±214.17 353.48±139.79* 35.9
* P＜0.05vs model group; #P＜0.05; ##P＜0.01vs normal group
Influence to malonaldehyde (MDA): body cell is subjected to free radical to attack degree: terpenoid＜EA-3＜phenolic acid＜50% ethanol＜EA-2＜insoral.Referring to table 2.
MDA mainly tests and reflects body inner lipid peroxidating degree, the indirect reaction body cell extent of damage.Cooperate with the mensuration of SOD, the height reflection body of SOD vigor is removed the ability of oxygen-derived free radicals, and the high mental retardation of MDA reflects the order of severity that body cell is attacked by free radical indirectly.Numerical value is high more, illustrates that body cell is subjected to the order of severity of free radical attack serious more.
Table 2, each position of Ramulus Euonymi are to the influence of malonaldehyde (MDA) (n=12, X ± S)
MDA content
Dosage (mg/kg) nmol/ml
Normal group/25.19 ± 3.57
Model group/31.90 ± 6.18
Insoral 110 39.29 ± 14.98
I 3.2 23.73±8.57
*
II 3 20.65±6.28
***
III 3 24.67±6.48
*
IV 1.5 22.76±5.15
**
V 1.5 27.20±7.48
* P＜0.05; * P＜0.01; * * P＜0.01vs model group
Influence to superoxide dismutase (SOD): remove ultra-oxygen anion free radical (O
2) ability: phenolic acid＞50% ethanol＞EA-3＞terpenoid＞EA-2, and insoral does not have this effect.Referring to table 3.
Diabetics is often removed the superoxide dismutase activity of free radical with the cylinder super-oxidation lipid level obviously raises low, and free radical increases the generation and the development that can increase the weight of diabetic complication.SOD plays crucial effects to the oxidation and the antioxidation balance of body, and the height reflection body of its vigor is removed the ability of ultra-oxygen anion free radical.
Table 3, each position of Ramulus Euonymi are to the influence of superoxide dismutase (n=12, X ± S)
Dosage SOD vigor
Group (mg/kg) (NU/ml)
Normal group/172.22 ± 69.82
Model group/148.32 ± 46.66
Insoral 110 135.61 ± 101.90
I 3.2 382.47±55.67***
II 3 191.94±97.80
III 3 465.81±53.64***
IV 1.5 250.66±96.69*
V 1.5 163.50±82.17
* P＜0.05; * P＜0.01; * * P＜0.01vs model group
Experiment shows: aforementioned 50% ethanol elution position and EA-2 of Ramulus Euonymi, and it is remarkable to reduce the blood glucose in diabetic mice effect, and using dosage is little, and terpenoid, phenolic acids and EA-3 are then to MDA, SOD influence obviously.Relatively comprehensive, the modern preparation of the Chinese medicine that hypoglycemic effective site of Ramulus Euonymi and effective ingredient are made, side effect significantly reduces and reduces, and still has suitable safety even make the patient take for a long time.Even more ideal is, only cures the symptoms, not the disease with insoral and compares, and Ramulus Euonymi Chinese medicine preparation of the present invention has the treating both the principal and secondary aspects of a disease function, can alleviate or suppress the generation and the development of diabetic complication.
Extraction, the separation method of the above-mentioned blood sugar lowering Chinese medicine composition of preparation of the present invention's research, favorable reproducibility can be used in suitability for industrialized production.
The specific embodiment
Instrument and reagent:
ZK-82A type vacuum drying oven, AE240 electronic analytical balance, the medical numerical control Ultrasound Instrument of KQ-100DE, constant water bath box, drawdown pump.7550 ultra-violet and visible spectrophotometers (Shanghai analytical tool factory), centrifugal settler (Shanghai Surgical Operation Equipment Factory).Agents useful for same is analytical pure.Column chromatography and tlc silica gel are Haiyang Chemical Plant, Qingdao's product.Polyamide chromatography and polyamide thin layer chromatography are Shanghai chemical reagent factory of Chinese Medicine group product.
Medical material and medicine:
The Ramulus Euonymi medical material is available from Nanjing medical material company, through being accredited as branch and the aliform appurtenance of Celastraceae plant Euonymus alatus (Thunb.) Sieb..
Glyburide is that Tianjin Pharmaceutical Co., Ltd of drug research institute produces lot number 9902311.
Insoral, Jintan City, Jiangsu Province pharmaceutical factory produces, lot number 9901202,20011201.
Alloxan, U.S. SIGMA company produces.
Glucose kit, Rong Sheng biotech company in Shanghai produces; Lot number: 20020503
MDA test kit, Nanjing build up bio-engineering research production; Lot number: 20020715
SOD test kit, Nanjing build up bio-engineering research production; Lot number: 20020725
Animal: Kunming mouse, male and female half and half, body weight 22～25g and 18～22g.Provide by China Medicine University's animal center.
Embodiment 1:
Get Ramulus Euonymi medical material 5kg, decoct 2 times with 6 times of water yields, each 2 hours, filtered while hot.Get extractum 250g after extracting solution concentrates, extractum is used petroleum ether, ethyl acetate, water saturated n-butanol extraction successively.Make it be divided into five positions, be respectively aqueous extract group (I), petroleum ether group (II), ethyl acetate group (III), n-butyl alcohol group (IV), the collection remaining group in back (V).
Each extract part is to the blood sugar reducing function of modeling mice:
(body weight 18～22g) is divided into seven groups at random by body weight, 10 every group to get 70 of healthy mices.Press the 0.2ml/10g gastric infusion, once a day, continuous 7 days.The treatment group is all by being equivalent to crude drug amount 4g/kg administration (I 400mg/Kg; II 1.00mg/Kg; III 17.20mg/Kg; IV 38.60mg/Kg; V344.00mg/Kg), matched group is given the 0.5%CMC-Na of equivalent, and the positive drug group is given glyburide (100mg/kg).Beginning fasting 2 hours (drinking-water is not limit) before the last administration, fasting blood sugar (FSG) is surveyed in administration after 2 hours.
The modeling experimental technique, referring to: Huang Min etc., different fasting time are to the influence of the serum insulin and the blood glucose of alloxan diabetes mouse model, China Medicine University's journal, 2001,32 (3): 221-223.
(body weight 22～25g) is raised fasting 16 hours (drinking-water is not limit) after 3 days to healthy mice, and (with water recently distilled preparation, concentration 1%, dosage: 200mg/kg), the normal raising after 72 hours, surveyed fasting blood sugar (FSG) to the lumbar injection alloxan.The mice of choosing FSG＞180mg/kg is a diabetic mice.
12 of first picked at random healthy mices are as normal group before the modeling.Choose 84 of diabetic mices after other mice modelings, be divided into 7 groups at random by fasting blood sugar.Each treatment group is by being equivalent to Ramulus Euonymi crude drug amount 4g/kg administration (I 400mg/Kg; II 1.00mg/Kg; III 17.20mg/Kg; IV 38.60mg/Kg; V344.00mg/Kg), normal group and model group are given the CMC-Na of equivalent 0.5%, and the positive drug group is given and insoral (110mg/kg).Administering mode is pressed 0.2ml/10g and is irritated stomach.Successive administration 7 days, last administration fasting in preceding 2 hours, two hours after administration is surveyed FSG.
Each extract part is to the blood sugar influence of healthy mice
By visible each extract part of Ramulus Euonymi of table 4 blood sugar reducing function and matched group comparison no significant difference P＞0.05 to healthy mice.
Each extract part of table 4. Ramulus Euonymi is to the blood sugar reducing function of healthy mice (n=12, X ± S)
| Group | Dosage (mg/kg) | FSG(mg/dl) |
| Normal group glyburide I II III IV V | / 100.00 400.00 1.00 17.20 38.60 334.00 | 45.99±14.59 25.06±11.78 ** 51.26±17.89 40.86±19.49 32.82±16.44 37.71±19.45 41.50±11.53 |
*P＜0.05vs normal group
Each extract part is to the influence of the chemical diabetes mouse blood sugar due to the alloxan
The ethyl acetate extract of Ramulus Euonymi (III), water extraction position (I), residual water position (V), extraction back blood sugar reducing function have remarkable meaning, and petroleum ether partly (II), n-butanol portion (IV) effect are not remarkable.Referring to table 5.Research proves that repeatedly water extraction position blood sugar reducing function is clear and definite, and ethyl acetate extract and the equivalence of water extraction position are so determine that ethyl acetate extract is further object of study.
Table 5, extract each position to chemical diabetes that alloxan caused
The influence of mouse blood sugar (n=12, X ± S)
| Group | Dosage (mg/kg) | Blood glucose value (mg/dl) | Blood glucose decline percentage rate (%) | |
| Before the administration | After the administration | |||
| Normal group model group insoral | / / 110.0 | 127.01±19.67 391.13±45.86 363.08±58.20 | 117.41±26.67 380.34±55.14 145.87±92.74 ***### | 7.6 2.8 59.8 |
| I II III IV V | 400.0 1.0 17.2 38.6 334.0 | 419.55±85.35 390.23±101.02 386.15±93.99 397.88±67.69 414.35±122.4 | 199.38±94.38 ***### 265.99±114.78 159.09±68.88 ***### 327.723±133.54 257.87±70.50 ## | 52.5 31.8 57.1 17.6 37.8 |
* *P＜0.01vs model group; ##P＜0.05 ###P＜0.01vs normal group;
In order further to prove conclusively its active site, we adopt polyamide column chromatography to the ethyl acetate extraction position.
Embodiment 2:
Each chemical constituent of ethyl acetate extract is to the influence of the chemical diabetes mouse blood sugar that alloxan caused.
The chemical constituent qualitative analysis is carried out at the ethyl acetate extraction position found that it mainly contains constituents such as terpenoid, steroidal, phenolic acid.Be divided into 30% ethanol component (III-1), 50% ethanol component (III-2), 95% ethanol component (III-3) behind the extraction separation and carry out pharmacodynamics relatively; Found that the hypoglycemic effect of III-2 component and water extraction position is suitable.See Table 6.
Table 6, isolating each component of ethyl acetate extract are to chemical diabetes that alloxan caused
The influence of mouse blood sugar (n=12, X ± S)
| Group | Dosage (mg/kg) | Blood glucose value (mg/dl) | Blood glucose decline percentage rate (%) | |
| Before the administration | After the administration | |||
| Normal group model group phenformin I III III-1 III-2 III-3 | / / 110.00 200.00 9.56 4.54 3.20 0.64 | 139.03±22.74 445.55±164.93 434.04±159.75 439.70±144.80 445.05±165.59 368.79±171.78 386.57±169.07 411.51±149.52 | 139.18±32.127 418.92±141.75 209.80±88.656 *** 241.21±114.34 *** 285.84±152.51 ** 368.22±115.01 246.24±150.61 ***Δ 300.00±158.23 | -0.11 5.9 51.7 45.1 35.8 0.1 36.3 27.1 |
*P＜0.05
* *P＜0.01vs model group; P＞0.05vs normal group
Take by weighing polyamide column (15-60 order) on the embodiment 1 gained 5 gram Ramulus Euonymi ethyl acetate extracts, water, 30% ethanol, 50% ethanol, 95% ethanol carry out column chromatography successively, and water and 30% component merge, and obtain three components.Be that water adds 30% ethanol component (III-1), 50% ethanol component (III-2) and 95% ethanol component (III-3).
Each component of ethyl acetate extract is to the blood sugar reducing function of diabetic mice:
Get ethyl acetate extract chemical constituent III-1, III-2, III-3 and Ramulus Euonymi extract part I, III, carry out the pharmacodynamics evaluation by the previous experiments method.Normal group and model group are given the CMC-Na of equivalent 0.5%, and the positive drug group is given and insoral (110mg/kg).
In view of the hypoglycemic activity optimum of ethyl acetate extract 50% ethanol component (III-2), we have further carried out the extraction separation of chemical constituent to this component.
Embodiment 3:
Get Sephadex LH-20 post on the extractum of III-2 component, use chloroform: methanol=1: 1 eluting, gained extractum are gone up 200-300 order silicagel column again, with the chloroform and the methanol gradient elution of different proportion, get compd E A-1～4, all ferric chloride reagent is reacting positive.
The crystallization of EA-1 colourless powder shape, mp.213-214 ℃.ESI-MS(m/z)：137[M-1]
-。
1H-NMR(300MHz，Acetone-d
6)δ9.62(1H，s，-COOH)7.88(2H，d，J＝8.8Hz，2-H and6-H)，6.89(2H，d，J＝8.8Hz，3-H and 5-H)；
13C-NMR(300MHz，Acetone-d
6)：168.27(-COOH)，163.27(C-4)，133.34(C-3andC-5)，123.33(C-1)，116.57(C-2andC-6)。Physicochemical data is consistent with the P-hydroxybenzoic acid of spectroscopic data and bibliographical information, so determine that this chemical compound is a P-hydroxybenzoic acid.
The EA-2 colourless crystallization, mp.ESI-MS (m/z): 153[M-1]
- 1H-NMR (300MHz, Acetone-d
6) δ .51 (1H, d, J=2.0Hz, 2-H), 7.44 (1H, dd, J=8.3,2.0Hz, 6-H), 6.88 (1H, d, J=8.3Hz, 5-H)
13C-NMR (300MHz, Acetone-d
6): 166.63 (COOH), 149.59 (C-4), 144.44 (C-3), 122.45 (C-6), 122.01 (C-1), 116.32 (C-2), 114.51 (C-5) physicochemical data is consistent with spectroscopic data and bibliographical information, so determine this chemical compound 3,4-resorcylic acid.
The EA-3 colourless crystallization, mp.ESI-MS (m/z): 167[M-1]
- 1H-NMR (500MHzAcetone-d
6) δ 7.61 (1H, dd, J=8.3,1.9Hz, 6-H), 7.58 (1H, d, J=1.9Hz, 2-H), 6.88 (1H, d, J=8.2Hz, 5-H), 3.91 (3H, s ,-OCH
3)
13C-NMR (500MHz, Acetone-d
6): 166.34 (COOH), 150.91 (C-4), 146.91 (C-3), 123.73 (C-6), 121.77 (C-1), 114.34 (C-2), 112.45 (C-5), 55.22 (OCH
3) physicochemical data is consistent with spectroscopic data and bibliographical information, so definite this chemical compound 3-methoxyl group-4-hydroxy benzoic acid.
The colourless needle of EA-4, mp.ESI-MS (m/z): 197[M-1]
- 1H-NMR(300MHz，Acetone-d
6)δ7.3(2H，s，2-H and 6-H)，3.87(6H，s.-OCH
3)；
13C-NMR(300MHz，Acetone-d
6)：168.32(-COOH)，149.00(C-3andC-5)，142.01(C-4)，122.20(C-1)，108.84(C-2 and C-6)，57.34(-OCH
3)。Physicochemical data is consistent with spectroscopic data and bibliographical information, so determine this chemical compound 3.5-dimethoxy-4 '-hydroxy benzoic acid.
Embodiment 4: get 5 kilograms of Ramulus Euonymis directly with the ethyl acetate backflow of 5 times (W/V) 2 times, gained extractum 15 grams are gone up polyamide column, with second alcohol and water gradient elution, get 50% ethanol elution position, reclaim solvent, get ethyl acetate extract 50% ethanol component.
Embodiment 5: get 5 kilograms of Ramulus Euonymis, boil 2 times with 6 times of (W/V) decoctings, concentrate back gained extractum with polyamide column on the wet method, with second alcohol and water gradient elution, get 50% ethanol elution position, reclaim solvent, with 50% ethanol elution position ethyl acetate extraction, get ethyl acetate extract.
Embodiment 6: get 5 kilograms of Ramulus Euonymis, with 50% alcohol reflux of 5 times (W/V) 2 times, gained extractum 300 grams, extract successively with petroleum ether, ethyl acetate, get ethyl acetate extraction part, last polyamide column (30-60 order), with second alcohol and water gradient elution, get 50% ethanol elution position, reclaim solvent, get ethyl acetate extract 50% ethanol component.
Claims (4)
1, a kind of hypoglycemic Chinese medicine composition is characterized in that active component makes with the following method:
1) get the Ramulus Euonymi medical material, water or 50% ethanol extraction concentrate extracting solution, obtain concentrate;
2) get the concentrate of water in the step 1) or 50% ethanol extract, use ethyl acetate extraction, concentrated extract obtains acetic acid ethyl ester extract, is the active component of described Chinese medicine composition;
Perhaps, get the concentrate of water in the step 1) or 50% ethanol extract, carry out chromatography with 15-60 order polyamide, use the aquiferous ethanol gradient elution, collect its ethanol elution position of 50%, reclaim solvent, with 50% ethanol elution position ethyl acetate extraction, concentrated extract obtains the mixture of terpenoid and phenolic acid compound, is the active component of described Chinese medicine composition.
2, the Chinese medicine composition of claim 1 is characterized in that: described acetic acid ethyl ester extract, further carry out chromatography with 15-60 order polyamide, and use the aquiferous ethanol gradient elution, collect its ethanol elution position of 50%.
3, a kind of hypoglycemic Chinese medicine composition is characterized in that active component makes with the following method:
1) gets the Ramulus Euonymi medical material, use ethyl acetate extraction, extracting solution is concentrated, obtain concentrate;
2) get the concentrate of acetic acid ethyl acetate extract in the step 1), carry out chromatography, use the aquiferous ethanol gradient elution with 15-60 order polyamide, collect its ethanol elution position of 50%, after concentrating, obtain the mixture of terpenoid and phenolic acid compound, be the active component of described Chinese medicine composition.
4, the application of the mixture of the active component terpenoid of claim 1 or 3 Chinese medicine composition and phenolic acid compound in preparation blood sugar lowering medicine.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 02138362 CN1241559C (en) | 2002-09-29 | 2002-09-29 | Chinese traditional medicine hypoglycemic agent and preparing process thereof |
| CNB2005100060677A CN1321637C (en) | 2002-09-29 | 2002-09-29 | Chinese medicinal hypoglycemic agent and preparation thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 02138362 CN1241559C (en) | 2002-09-29 | 2002-09-29 | Chinese traditional medicine hypoglycemic agent and preparing process thereof |
Related Child Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB2005100060677A Division CN1321637C (en) | 2002-09-29 | 2002-09-29 | Chinese medicinal hypoglycemic agent and preparation thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1401318A CN1401318A (en) | 2003-03-12 |
| CN1241559C true CN1241559C (en) | 2006-02-15 |
Family
ID=4749436
Family Applications (2)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB2005100060677A Expired - Fee Related CN1321637C (en) | 2002-09-29 | 2002-09-29 | Chinese medicinal hypoglycemic agent and preparation thereof |
| CN 02138362 Expired - Fee Related CN1241559C (en) | 2002-09-29 | 2002-09-29 | Chinese traditional medicine hypoglycemic agent and preparing process thereof |
Family Applications Before (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB2005100060677A Expired - Fee Related CN1321637C (en) | 2002-09-29 | 2002-09-29 | Chinese medicinal hypoglycemic agent and preparation thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (2) | CN1321637C (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110464717B (en) * | 2019-09-23 | 2022-02-18 | 张建国 | Application of 2, 4-dihydroxybenzoic acid in preparation of blood sugar reducing medicine |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB2000025A (en) * | 1977-05-14 | 1979-01-04 | Pfizer Ltd | Nitroimidazole formulations |
| CN1144098A (en) * | 1994-01-15 | 1997-03-05 | 河南省密县科学技术研究所 | Oral liquor with functions of reducing blood sugar and invigorating kidney |
| CN1212889A (en) * | 1997-09-29 | 1999-04-07 | 全龙浩 | Traditional Chinese medicine component for improving immunocompetence and having effect on multiple epidemic diseases |
-
2002
- 2002-09-29 CN CNB2005100060677A patent/CN1321637C/en not_active Expired - Fee Related
- 2002-09-29 CN CN 02138362 patent/CN1241559C/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| CN1321637C (en) | 2007-06-20 |
| CN1679529A (en) | 2005-10-12 |
| CN1401318A (en) | 2003-03-12 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN1237961C (en) | Use of stilbene compounds in preparing medicaments for treating and preventing diabetes or diseases associated with retrovirus | |
| CN1754541A (en) | Steroid saponin pharmaceutical composition and its preparation method and uses | |
| CN1931270A (en) | Sobering up and liver protecting Chinese medicine prepn and its prepn process | |
| CN1850836A (en) | Celosia argentea suponin compound and its pharmaceutical use | |
| CN1840166A (en) | Modern Chinese medicinal oral liquid of 'Wen Dan Tang' and preparation method thereof | |
| CN101032504A (en) | Application of liquiritin in medicines | |
| CN1224383C (en) | Blood sugar reducing compound | |
| CN1817898A (en) | Use of anti-inflammatory medicine for scheelite total saponin and its saponin compound | |
| CN1733054A (en) | Dogwood fruit extract and its preparation process | |
| CN1241559C (en) | Chinese traditional medicine hypoglycemic agent and preparing process thereof | |
| CN102579530A (en) | Preparation method of aralia taibaiensis total saponin having diabetes mellitus resisting effect and medicament | |
| CN103432420B (en) | A kind of Chinese medicine composition for the treatment of diabetes and preparation method thereof and detection method | |
| CN101045077A (en) | Preparating method of oral solid preparation of fenugreek and its use | |
| CN1218701C (en) | Use of spirosterol type steroid saponin in preparing medicine for treating cardio-cerebral vascular disease | |
| CN1853688A (en) | Chinese medicinal preparation for treating heart cerebrovascular disease and ischemic apoplexia and making method thereof | |
| CN1895540A (en) | Medicinal composition for treating cardiovascular disease, its making method and use | |
| CN106083842A (en) | The Preparation method and use of 9 substituted double-functional group berberinc derivates | |
| CN111920799A (en) | Kulecuo effective component composition and preparation method and application thereof | |
| CN101032534A (en) | Method of preparing jiubiying total saponins and the application thereof | |
| CN1548142A (en) | Medicine for improving sugar tolerance and treating diabetes and its prepn process | |
| CN1539844A (en) | 2,3,5,4'-tetrahydroxy diphenyl ethylene-2-1-beta-D-glucoside | |
| CN1176939C (en) | Fructus gleditsiae total saponin, its preparation method and use in preparation of medicine | |
| CN1590396A (en) | Picrorhiza total glycoside extract and its application in preparation of liver disease medicine and preparation method | |
| CN106146488A (en) | The Preparation method and use of 9 substituted double-functional group berberinc derivates | |
| CN1543968A (en) | Drug prepared by mulberry bark extract |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C17 | Cessation of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20060215 Termination date: 20100929 |