A kind of preparation method and pharmacological action thereof of Mangiferin six fourth esterified derivatives
Technical field
The present invention relates to, take Mangiferin as starting raw material, prepare a kind of method of new organic compound, and this compound is in anti-inflammatory, pharmacological action aspect hypoglycemic.
Technical background
Many Chinese medicinal materialss are as fruit, leaf, the bark of Anacardiaceae plant mango (Mangifera indica L.), rhizome, the over-ground part of the liliaceous plant wind-weed (Anemarrhena asphodeloides Bge.), flower, the leaf etc. of irides blackberry lily (Belamcanda chinensis (L.) DC.) all contain Mangiferin (Mangiferin).Mangiferin is a kind of glucose C-glucosides (molecular formula C of the Xanthone of having structure
19h
18o
11, molecular weight 422.34).
Mangiferin molecule has larger two dimensional structure, poor (the average solubleness < 1mgmL in the buffered soln of pure water and pH 1.3~7.4 of solubleness
-1), and be 630mg through conversion people's oral effective dose, dissolve the GI liquid volume > 250mL that single dose Mangiferin needs, according to biopharmacy categorizing system (Biopharmaceutics classification system, BCS) definition, Mangiferin belongs to a kind of poorly soluble medicine.In addition, Mangiferin is partition ratio (Oil/water partition coefficient in n-butanol-water system, P) very little (pH1~4 o'clock, P value is about 2.3, and in the time that pH value is greater than 6.86, P value is only 0.01), conventionally P value and stomach and intestine permeability good relationship, P value was at 100~1000 o'clock, and Passive intake is better, inferred that Mangiferin stomach and intestine permeability is poor.Moreover Mangiferin intestinal absorption effect in Mice Body is poor, confirm that Mangiferin permeability is poor.The character of the comprehensive above-mentioned three aspects: of Mangiferin, the low Determination of oil-water partition coefficient of Mangiferin may be directly related with its low bioavailability in Mice Body, according to biopharmacy categorizing system (BCS) principle, Mangiferin can classify as BCS the 4th class medicine (low-solubility-low permeability).
In order to solve the absorption problem of BCS the 4th class medicine (low-solubility-low permeability), it is derivative that medicine is carried out esterification by the large more options of Chinese scholars, forms fat-soluble high derivative, thereby improve the cross-film permeability of medicine, the absorption that improves medicine.
Summary of the invention
First technical problem that the present invention will solve is: adopt chemical synthesis process, prepare Mangiferin six fourth esterified derivatives, its structural formula is as follows:
Preparation method and step are as follows: under continuous stirring and cooling conditions, in butyryl oxide, drip high-concentration inorganic acid (the wherein mixture of one or more: mass percent concentration is not less than 90% H
2sO
4, be not less than 30% HCl, be not less than 90% H
3pO
4), obtain solution A; Under continuous stirring and cooling conditions, Mangiferin is dissolved in A, obtain solution B; B is incubated to 18-72 hour in 60-90 ℃ of environment, obtains reaction liquid C; Under continuous agitation condition, by direct C impouring water; Leaching insolubles, obtains reaction product D; By D silica gel mixed sample, put on the dry capital of silica gel, with sherwood oil: chloroform: acetone gradient elution, at sherwood oil: chloroform: acetone=10: obtain crystallization in 7: 3 flow points, be Mangiferin six fourth esterified derivatives.
Second technical problem that the present invention will solve is: the structural identification of mangiferin derivatives.Infrared, ultraviolet to derivative, hydrogen spectrum, carbon spectrum, two-dimensional spectrum, mass spectrum (low, high resolving power) are resolved, and have confirmed its structure.Characterization of compound data: off-white color crystal, mp 158-160 ℃;
1h NMR (CDCl
3, 600Hz) and d:7.91 (1H, s, H-8), 7.31 (1H, s, H-5), 6.84 (1H, s, H-4), 5.30~3.89 (protons on sugar), 2.52~1.34 (24H, q), 1.11 (3H, t), 1.03 (3H, t), 1.01 (3H, t), 0.95 (3H, t), 0.90 (3H, t), 0.86 (3H, t).
13C?NMR(CDCl
3,125Hz)d:161.2(C-1),112.3(C-2),158.8(C-3),93.4(C-4),153.1(C-10a),103.8(C-5),147.5(C-6),147.5(C-7),112.3(C-8),120.8(C-8a),179.9(C-9),153.1(C-9a),102.3(C-4a),73.3(C-1′),70.6(C-2′),73.5(C-3′),67.4(C-4′),76.6(C-5′),61.0(C-6′),36.4,36.1,36.0,36.0,35.9,35.6(6CH
2),18.6、18.5、18.4、18.4、18.2、18.1(6CH
2),14.1、13.9、13.8、13.8、13.8、13.6(6CH
3),173.6、173.3、172.8、172.0、171.1、171.3(6C=O)。EI-MS:842[M]
+,758,702,646,592,536,523(100),467,411,354。HR-MS:842.3368,Calcd.842.3361,Calcd.for?C
43H
54O
17。
The 3rd problem that the present invention will solve is: the pharmacological action of clear and definite Mangiferin six fourth esterified derivatives.Adopt dimethylbenzene induced mice auricle edema model research anti-inflammatory drug effect, result shows, derivative has significant anti-inflammatory action; Adopt the diabetic mice model research hypoglycemic drug effect of tetraoxypyrimidine induction, result shows, derivative has significant hypoglycemic activity, and diabetic mice is had to obvious result for the treatment of.
Embodiment
The following example is used for illustrating the present invention, is not any restriction to protection domain of the present invention.
Embodiment 1: prepare Mangiferin six fourth esterified derivatives
In water-bath, operate, under continuous agitation condition, in 250ml butyryl oxide, drip 1.5ml 98%H
2sO
4, after dripping off, then add 10g Mangiferin; This solution is placed in to 40 ℃ of environment and is incubated, react 24 hours, frequently stir Mangiferin is all dissolved during this time; React complete, under continuous agitation condition, by direct reaction solution impouring water; Leaching insolubles, obtains reaction product; By reaction product silica gel mixed sample, put on the dry capital of silica gel, first use chloroform wash-out, then with chloroform: ethyl acetate: acetone=20: 1: 1 wash-out, then use chloroform: ethyl acetate: acetone=7: 2: 1 wash-outs, collect flow point, volatilize, obtain coarse crystallization; Coarse crystallization recrystallizing methanol 2-3 time, obtains Mangiferin six fourth esterified derivatives.
Embodiment 2: prepare Mangiferin six fourth esterified derivatives
In water-bath, operate, under continuous agitation condition, in 250ml butyryl oxide, drip 4ml 36%HCl, after dripping off, then add 10g Mangiferin; This solution is placed in to 50 ℃ of environment and is incubated, react 12 hours, frequently stir Mangiferin is all dissolved during this time; React complete, under continuous agitation condition, by direct reaction solution impouring water; Leaching insolubles, obtains reaction product; By reaction product silica gel mixed sample, put on the dry capital of silica gel, first use chloroform wash-out, then with chloroform: ethyl acetate: acetone=20: 1: 1 wash-out, then use chloroform: ethyl acetate: acetone=7: 2: 1 wash-outs, collect flow point, volatilize, obtain coarse crystallization; Coarse crystallization recrystallizing methanol 2-3 time, obtains Mangiferin six fourth esterified derivatives.
Embodiment 3: anti-inflammatory pharmacological evaluation and result
Mouse auricle swelling test is got 40 of mouse, is divided at random 4 groups.If Normal group, Asprin group (0.2g/kg), the Mangiferin six large small dose group of fourth esterified derivative (0.4g/kg, 0.2g/kg), every day gastric infusion 1 time, Normal group gives equal-volume distilled water, continuously 7d.After last administration after 0.5h, get 20 μ l dimethylbenzene with microsyringe and drip and be applied to mouse right ear, after 15min by drawing cervical vertebra to put to death mouse.The punch tool that is 6mm with bore immediately, draw materials along the punching of mouse left and right auricle same area, then use ten thousand/electronic balance to weigh respectively, using the difference of two auricle weight (mg) as swelling level index, swelling=(auris dextra weight-left ear weight)/auris dextra weight, the results are shown in Table 1.
Test-results shows, Mangiferin six fourth esterified derivatives have significant antiphlogistic effects.
Table 1 anti-inflammatory test-results
Note: through t check, with Normal group comparison: * P < 0.05; * P < 0.01.
Embodiment 4: hypoglycemic pharmacology experiment and result
Tetraoxypyrimidine hyperglycemia model mouse hypoglycemic test: get 90 of male KM mouse, body weight (25 ± 2g) after adaptability raising 1W.Separate at random 10 for Normal group by body weight, remain 80 mouse for experiment treatment group.Water 12h is can't help in all mouse fasting, and except Normal group injection equal-volume NS, all the other each caudal veins are injected the tetraoxypyrimidine NS solution that freshly prepared concentration is 3.5mg/ml (mouse dosage 35mg/kg, administration capacity 0.1ml/10kg).In the modeling of tail vein injection tetraoxypyrimidine, after 5 days, fasting 3h tail venous blood sampling, with the stable rapid blood sugar instrument test of three promises blood sugar.Except normal group, all the other mouse are got the diabetic mice of blood glucose value between 11~26mmol/L, supply test by blood sugar height again random packet.Each mouse administration every day 1 time, gavage volume is unified is each 0.2ml/10g body weight, and normal group, model group are to equal-volume pure water.Be 14 days the course for the treatment of, in administration 0 day, 14 days surveys blood glucose value, more each treated animal blood glucose value and blood sugar decline percentage.Blood glucose value × 100% before blood sugar decline percentage %=(the rear blood glucose value of blood glucose value-experiment before experiment)/experiment.
Animal experiment shows, N1,N1-Dimethylbiguanide positive drug group group and the large, medium and small dosage group of Mangiferin six fourth esterified derivative all can obviously reduce the blood sugar of the diabetic mice of tetraoxypyrimidine induction, and the diabetes of animal model are had to significant result for the treatment of.
The hypoglycemic test-results of table 2
Note: through t check, with model group comparison, * * p < 0.01.