CN101874825B - Medicinal composition for treating ulcerative colitis and preparation method thereof - Google Patents
Medicinal composition for treating ulcerative colitis and preparation method thereof Download PDFInfo
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- CN101874825B CN101874825B CN200910059160.2A CN200910059160A CN101874825B CN 101874825 B CN101874825 B CN 101874825B CN 200910059160 A CN200910059160 A CN 200910059160A CN 101874825 B CN101874825 B CN 101874825B
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Abstract
The invention provides application of a gastric medicament release system prepared from astragalus or active ingredients thereof and a colon positioning medicament release system prepared from lightyellow sophora root or active ingredients thereof in the preparation of combined medicaments for treating ulcerative colitis, and also provides a medicinal composition for treating the ulcerative colitis, which is a preparation prepared by mixing the gastric medicament release system prepared from the astragalus or active ingredients thereof and the colon positioning medicament release system prepared from the lightyellow sophora root or active ingredients thereof. Through the comparative experimental study of medicinal effect of different administration modes of each component, compared with two preparation modes of total formula colon administration and total formula oral administration, a preparation prepared by combining the oral administration of astragalus polysaccharide and saponin with the enteroclysis of matrine and oxymatrine can resist rat colon tissue pathological injury caused by trinitro-benzene-sulfonic acid obviously, improve apparent physical signs such as hematochezia, diarrhoea, anorexia, indolence and the like of animals and play a role by regulating the peripheral blood TNF-alpha level and inflammatory reactions in colon tissue, has obvious curative effect and can both regulate the integral immunologic function and resist the local inflammatory reactions in colon.
Description
Technical field
The present invention relates to colon-specific drug delivery system prepared by stomach release system prepared by the Radix Astragali or its effective ingredient and Radix Sophorae Flavescentis or its effective ingredient purposes in the combination medicine of preparation treatment ulcerative colitis.
Background technology
Ulcerative colitis is a kind of former, and non-specific colon chronic disease is only invaded mucosa and the tela submucosa of colon under colonoscope as seen, and has rotten to the corn and shallow table ulcer.At present by the raw material Radix Astragali and Radix Sophorae Flavescentis compatibility treatment ulcerative colitis, existing Patents or bibliographical information, as: number of patent application: 200410040960.7, denomination of invention: a kind of pharmaceutical composition for the treatment of ulcerative colitis and its production and use, this Patent Application Publication a kind of pharmaceutical composition for the treatment of ulcerative colitis, it is that containing by the Radix Astragali, Radix Sophorae Flavescentis is the medicament that raw material is prepared from, and the present invention also provides preparation method and the purposes of this pharmaceutical composition.Drug regimen raw material refining formula of the present invention, drug dose is little, material base is clear and definite, determined curative effect, toxic and side effects are little; Colon targeting preparation of the present invention has dosage form novelty, medicine and is directly positioned target area (colon), makes target area drug level higher than other normal structure, reduces systemic side effects, and taking convenience, dose is little, material base is clear and definite, curative effect is more remarkable.The peaceful prescription of the intestinal research of healing of Chinese medicine new compound, medicine Leader the 27th the 3rd phase of volume of March in 2008, adopt the optimization experiment of trinitro-benzene-sulfonic acid ulcerative colitis in rats (ulcerative colitis, UC) model to heal the intestinal peaceful prescription side of tearing open and best proportioning; Adopt two kinds of UC rat models to compare the more peaceful drug effect of intestinal of decoction pieces compatibility and active component compatibility.Result tentatively determines that more the peaceful flavour of a drug of intestinal consist of: the Radix Astragali, each 15g of Radix Sophorae Flavescentis; The side of tearing open experiment shows, two medicines share curative effect and are better than alonely, and take the Radix Astragali: Radix Sophorae Flavescentis is, curative effect is best at 1: 1 o'clock; The curative effect no significant difference of decoction pieces compatibility and active component compatibility, confirm that Radix Astragali saponin, astragalus polysaccharides, matrine, oxymatrine are the material base of the party's onset, and determined that more the intestinal peaceful day takes recipe quantity and be: astragalus polysaccharides 239mg, oxymatrine 150mg, matrine 50mg, Radix Astragali saponin 25mg.
In sum, existing preparation is that all employing is colon locating administrated by the Radix Astragali, Radix Sophorae Flavescentis active component, and because material composition is complicated, can this kind of form of medication reach optimum medicine efficacy, for a person skilled in the art, is unforeseen.
Summary of the invention
Technical scheme of the present invention has been to provide colon-specific drug delivery system prepared by stomach release system prepared by a kind of Radix Astragali or its effective ingredient and Radix Sophorae Flavescentis or its effective ingredient purposes in the combination medicine of preparation treatment ulcerative colitis, and another technical scheme of the present invention has been to provide a kind of pharmaceutical composition for the treatment of ulcerative colitis.
The invention provides colon-specific drug delivery system prepared by stomach release system prepared by the Radix Astragali or its effective ingredient and Radix Sophorae Flavescentis or its effective ingredient purposes in the combination medicine of preparation treatment ulcerative colitis.
Wherein, the weight proportion of the described Radix Astragali and Radix Sophorae Flavescentis is: Radix Astragali 2-6 part, Radix Sophorae Flavescentis 3-7 part.
Further preferably, the described Radix Astragali and the weight proportion of Radix Sophorae Flavescentis are:
The weight proportion of the Radix Astragali and Radix Sophorae Flavescentis is: 4 parts of the Radixs Astragali, 4 parts of Radix Sophorae Flavescentiss.
Wherein, the effective ingredient of the described Radix Astragali is: Radix Astragali saponin extract, astragalus polysaccharide extract; The effective ingredient of described Radix Sophorae Flavescentis is Radix Sophorae Flavescentis total alkaloids, and its weight proportion is:
Radix Astragali saponin extract 10-43 part, astragalus polysaccharide extract 5-40 part, Radix Sophorae Flavescentis total alkaloids 20-70 part.
Wherein, in described Radix Sophorae Flavescentis total alkaloids, contain matrine and oxymatrine; The weight proportion of Radix Astragali saponin extract, astragalus polysaccharide extract, matrine and oxymatrine is:
Radix Astragali saponin extract 10-43 part, astragalus polysaccharide extract 5-30 part, matrine 5-18 part, oxymatrine 15-52 part.
The weight proportion of further preferably, described Radix Astragali saponin extract, astragalus polysaccharide extract, matrine and oxymatrine is:
13.3 parts of astragaloside extracts, 39.8 parts of astragalus polysaccharide extracts, 10.2 parts of matrines, 30.6 parts of oxymatrines.
Wherein, the percentage composition that contains astragaloside in described Radix Astragali saponin extract is 5%-10%, and the percentage composition of total saponins is 40%-73.61%; Polyoses content 40%-70% in astragalus polysaccharide extract; The percentage composition that contains total alkaloids in Radix Sophorae Flavescentis total alkaloids is 70%.
The present invention also provides a kind of combination medicine for the treatment of ulcerative colitis, the unit formulation that it comprises different size, the colon-specific drug delivery system of preparing to stomach release system prepared by the medicine Radix Astragali or its effective ingredient and Radix Sophorae Flavescentis or its effective ingredient for simultaneously, respectively or successively, and pharmaceutically acceptable carrier.
Combination medicine described in the present invention can include but not limited to stomach release system and colon-specific drug delivery system, be mixed and made into a compound preparation, or stomach release system and colon-specific drug delivery system mix as being placed in a medicine box or medicine packaging apparatus and use, or stomach release system is placed in different packings and uses together from colon-specific drug delivery system.
The present invention can be in identical or different pharmaceutical composition administration simultaneously, or can sequential administration.If sequential administration, lose to interval time of the second active component and should not make the benefit of the coordinating effect that active component associating can produce.No matter it is also understood that it is simultaneously administration or order administration, stomach release system and colon-specific drug delivery system can be individually dosed or with its any cooperative programs administration.
Wherein said stomach release system is to be prepared from micropill by the raw material of following weight proportioning and adjuvant: astragalus polysaccharide extract 199-597 part, Radix Astragali saponin extract 68-204 part, microcrystalline Cellulose 477.5-1432.5 part, micropowder silica gel 93-279 part, carboxymethyl starch sodium 53-159 part, coating 44.5-133.5 part; Described colon-specific drug delivery system is to be prepared from by the raw material of following weight proportioning and adjuvant: Radix Sophorae Flavescentis total alkaloids extract 102-306 part, microcrystalline Cellulose 377.5-1132.5 part, carboxymethyl starch sodium 30.5-91.5 part, coating 306-612 part.
Wherein, described stomach release system is that raw material and adjuvant by following weight proportioning is prepared from micropill: 398 parts of astragalus polysaccharide extracts, 136 parts of Radix Astragali saponin extracts, 955 parts of microcrystalline Cellulose, 186 parts of micropowder silica gels, 106 parts of carboxymethyl starch sodium, 89 parts of coatings; Described colon-specific drug delivery system is to be prepared from by the raw material of following weight proportioning and adjuvant: 204 parts of Radix Sophorae Flavescentis total alkaloids extracts, 755 parts of microcrystalline Cellulose, 61 parts of carboxymethyl starch sodium, coating 306-612 part.
Further preferably, the coating described in described stomach release system is Eudragit E100, Eudragit I or other gastric solubleness coating pre-mixing agents; The coating material of described colon-specific drug delivery system is: Eudragit S100, internal layer bag Eudragit E100 outer bag Eudragit L100 or internal layer bag EudragitE100, middle level involucrum polysaccharide, outer bag Eudragit L100.
The present invention also provides a kind of method of preparing this pharmaceutical composition, and it comprises the steps:
A, get the Radix Astragali or its effective ingredient, Radix Sophorae Flavescentis or its effective ingredient;
B, the Radix Astragali or its effective ingredient are prepared into stomach release system; Radix Sophorae Flavescentis or its effective ingredient are prepared into colon-specific drug delivery system mixing;
C, two parts are mixed to the pharmaceutically conventional preparation that adds that pharmaceutically acceptable adjuvant or complementary composition be prepared into.
Stomach release system of the present invention (or stomach release unit), refers to that medicine enters after stomach, shorter time release fast under sour environment, thus guarantee that medicine can be intestinal absorption rapidly, and the release that can guarantee medicine dosage form completely.Colon-specific drug delivery system (or colon positioning release unit) refers to proper method, avoid discharging medicine at Stomach duodenum, jejunum and ileum front end, but discharge a kind of drug-supplying system that medicine is brought into play local and whole body therapeutic effect after being transported to ileocecus, be a kind of preparation that is positioned at colon drug delivery.
The advantage of medicine of the present invention, is embodied in three aspects:
1. preparation mentality of designing is innovated the complexity based on disease pathogenesis and compound medicine character, fully holding colitis and the Chinese medicine of the peaceful raw material prescription of intestinal of healing, on the basis of modern medicine relevant information, adopt polynary release technology to process the cut the garment according to the figure preparation of formula of the peaceful side's Chinese medicine of intestinal more, to give full play to characteristic separately, and around the wholistic therapy of disease and each preparation unit is combined, form an organic modern Chinese medicine compound recipe medicine-releasing system, both inherited the marrow of Chinese medical theory and practice, possesses again modern preparation stabilization, controlled, feature easily.
2. because said preparation can effectively regulate whole immunologic function, the damage of colon local inflammation can be resisted again, treating both the principal and secondary aspects of a disease can be played, the dual characteristic that whole part is taken into account.Be used for the treatment of that light medium-sized UC can improve rapidly diarrhoea, the symptom such as have blood in stool, can regulate again patient's body constitution, regulate the inflammation state of colon intestinal, can prevent its recurrence.
3. easy to use.Tcm clinical practice treatment UC, generally adopts two kinds of modes of oral decoction and coloclysis to treat, and administration is frequent, complicated operation, and administration time is longer, and it is very inconvenient to treat.Adopt said preparation, directly common oral, both can realize the therapeutic purposes of clinical two kinds of administering modes.
Drug effect contrast experiment by each component different modes of administration studies, find and full reef knot enteral administration, two kinds of preparation ways of full side's oral administration are compared, more the peaceful astragalus polysaccharides of intestinal, saponin oral liquor and matrine, the combination of oxymatrine coloclysis, can significantly resist the rat colon histopathology damage that trinitro-benzene-sulfonic acid causes, improving animal has blood in stool, diarrhoea, anorexia, the outward appearance signs such as lazyness is moving, and by regulating inflammatory reaction in peripheral blood TNF-alpha levels and colon to play a role, curative effect is the most remarkable, both the whole immunologic function of scalable, can resist colon local inflammatory response again.
Obviously, according to foregoing of the present invention, according to ordinary skill knowledge and the customary means of this area, not departing under above-mentioned basic fundamental thought prerequisite of the present invention, can also make modification, replacement and the change of other various ways.
The specific embodiment of form, is described in further detail foregoing of the present invention again by the following examples.But this should be interpreted as to the scope of the above-mentioned theme of the present invention only limits to following example.All technology realizing based on foregoing of the present invention all belong to scope of the present invention.
Accompanying drawing explanation
Fig. 1 compound recipe medicine-releasing system of the present invention concrete technology route
The in vivo release curve of tri-batches of Radix Astragali micropills of Fig. 2
The in vivo release curve of Fig. 3 sophora root pellet
The specific embodiment
The preparation of embodiment 1 medicine of the present invention
1, the preparation of Radix Astragali stomach release unit:
Got Radix Astragali saponin extract 2.5g, astragalus polysaccharide extract 7.5g, microcrystalline Cellulose 18g, micropowder silica gel 3.5g, the carboxymethyl starch sodium 2g of No. six sieves of pharmacopeia (100 orders, 150 μ m), powder, mix, appropriate 50% ethanol of take is prepared soft material as wetting agent, extrudes, round as a ball, obtains micropill.Adopt the molten coating of Eudragit E100 alcohol, its coating prescription is Eudragit E100 10g; Pulvis Talci 5g; PEG6000 1g; Water 2g; Ethanol 140g.Eudragit E100 is dissolved in to about 70g ethanol; PEG6000 is water-soluble; Pulvis Talci adds in residue ethanol, high speed homogenize 20 minutes; PEG6000 solution and Pulvis Talci suspension are added in Eudragit E100 solution, stir evenly, obtain coating solution.Adopt fluidized bed coating technology, micropill is carried out to coating, make to increase weight 5%, obtain.
2, the preparation technology of Radix Sophorae Flavescentis colon positioning release unit:
The preparation of 2.1 sophora root pellets: got No. six sieve (100 orders of pharmacopeia, 150 μ m) Radix Sophorae Flavescentis total alkaloids 10g, microcrystalline Cellulose 37g, carboxymethyl starch sodium 3g, mix, with the appropriate moistening of water, prepare soft material, under the condition of speed 25Hz, soft material is extruded, under the condition of round as a ball speed 60Hz, rolling 4min, obtains micropill.The micropill of preparation is adopted respectively to following three kinds of mode coatings:
2.2pH responsive type single coats technique: get Eudragit S100 10g, Pulvis Talci 2.5g, triethyl citrate 1g, the 95% ethanol 170ml of take is dissolution with solvents, be mixed with the mixed liquor that concentration is about 8% (W/V), after stirring is spent the night, coating solution is crossed 200 mesh sieves and is filtered, 35 ℃ of inlet temperature; Materialization pressure 1.32～1.45bar; Fluidisation pressure 0.29～0.39bar; Feed liquor speed 20～69; 28 ℃ of temperature of charge; 75～80 ℃ of delivery temperatures; Under supply gas pressure 0.56mpa～0.66mpa condition, coating makes to increase weight 30%, takes out coated micropill, adds 1% Pulvis Talci, is dried 2h and get final product in 40 ℃.
2.3pH responsive type double-layer coatings technique: internal layer coating: adopt the molten coating prescription of Eudragit E100 alcohol: Eudragit E100 10g; Triethyl citrate 1g; 80% ethanol 140mL.10g EudragitE100 is dissolved in to about 70mL ethanol, and triethyl citrate adds in residue ethanol, and triethyl citrate solution is added in Eudragit E100 solution, after stirring is spent the night, crosses 200 mesh sieves and filters.Adopt fluidized bed coating, inlet temperature: 35 ℃, materialization pressure: 1.3～1.45bar, fluidisation pressure: 0.29～0.5bar, feed liquor speed: 19～59, temperature of charge: 28 ℃, delivery temperature: 80 ℃ of left and right, supply gas pressure: 0.68mpa left and right.Coating makes to increase weight 17%, takes out coated micropill, adds 1% Pulvis Talci, is dried 2h and get final product in 40 ℃.
Outer coating: adopt the molten coating prescription of Eudragit L100 alcohol: Eudragit L100 10g; Triethyl citrate 1g; 95% ethanol 140mL.Eudragit L100 is dissolved in to about 70mL ethanol, and triethyl citrate adds in residue ethanol, and stirring adds triethyl citrate solution in EudragitE100 solution after spending the night, and stirs evenly, and crosses 200 mesh sieves and filters.The same method is carried out outer coating to wrapping the micropill of internal layer clothing, and coating makes to increase weight 15%, takes out coated micropill, adds 1% Pulvis Talci, is dried 2h and get final product in 40 ℃.
2.4pH sensitivity-enzymatic is sent out dual control coating: internal layer coating: adopt the molten coating of pH responsive type double-layer coatings technique internal layer Eudragit E100 alcohol, with method coating, make to increase weight 10%, take out coated micropill, add 1% Pulvis Talci, be dried 2h and get final product in 40 ℃.
Middle level coating: get chitosan and add 1% acetate dissolution, be mixed with concentration and be 1% solution, then add the Pulvis Talci of polymer dry weight 100%, 10% dibutyl phthalate, stir spend the night after coating solution cross 200 mesh sieves.To wrapping the coated micropill of Eudragit E100 clothing layer, carry out secondary coating, coating makes micropill weightening finish 30%, take out coated micropill, in 2% aqueous alkali, soak 30min, washing is extremely neutral rapidly with clear water for micropill, blot immediately surperficial excessive moisture, add 1% Pulvis Talci to mix, standby at 40 degree oven drying 12h.
Outer coating: adopt the molten coating preparation of pH responsive type double-layer coatings technique internal layer Eudragit L100 alcohol coating solution, in fluid bed, Eudragit E100 and chitosan double-layer coatings micropill are carried out to coating for the third time, coating makes to increase weight 20%, take out coated micropill, add 1% Pulvis Talci, in 40 ℃, dry 2h obtains coated micropill.
3. the mixing of liang release unit
By stomach release unit coated micropill and colon positioning release coated micropill according to dosage ratio mix, minute encapsulated, obtain pharmaceutical preparation of the present invention.
The preparation of embodiment 2 medicines of the present invention
1, stomach release unit preparation prescription
Radix Astragali coated micropill
Astragalus polysaccharide extract 398mg (containing polysaccharide 355mg) Radix Astragali saponin extract 136mg (containing saponin 100mg)
Microcrystalline Cellulose 961mg micropowder silica gel 187mg
Carboxymethyl starch sodium 107mg coating 90mg
Be total to 1879mg
2, colon positioning release unit preparation prescription
2.1pH responsive type single coats micropill
Matrine extract 51mg (matrine 50mg) oxymatrine extract 153mg (oxymatrine 150mg)
Microcrystalline Cellulose 755mg carboxymethyl starch sodium 61mg
Coating 306mg
Be total to 1326mg
2.2pH responsive type double-layer coatings micropill
Matrine extract 51mg oxymatrine extract 153mg
Microcrystalline Cellulose 755mg carboxymethyl starch sodium 61mg
Coating 326mg
Be total to 1346mg
2.3pH sensitivity-enzymatic is sent out dual control coated micropill
Matrine extract 51mg oxymatrine extract 153mg
Microcrystalline Cellulose 755mg carboxymethyl starch sodium 61mg
Coating 612mg
Be total to 1632mg
According to three of colon positioning release unit kinds of coating schemes, more the daily dose of the peaceful stomach-colon of intestinal subrelease capsule is slightly variant, is respectively 3.21g, 3.23g, 3.52g, be sub-packed in 9 capsules, every heavily about 0.36g, 0.36g, 0.39g respectively, every day three times, each 3.
Embodiment 3 excipient substance of the present invention and preparation technology's screening test
One, the preparation of medicine stomach-colon subrelease capsule of the present invention
Medicine of the present invention is divided into two parts, wherein be prepared into can be in the stomach preparation unit of release fast for Radix Astragali saponin, polysaccharide, matrine, oxymatrine are prepared into the preparation unit with colon positioning release performance, then the compound recipe medicine-releasing system that two preparation unit Hybrid assemblings are become to comprise two kinds of drug release behaviors.
Concrete technology route is shown in Fig. 1.
1 instrument and reagent:
1.1 instrument
Granulator for spherical particulate (East China University of Science and technology Chemical Engineering Machine Insitute)
Friability monitor (FT-2000 type, Tianjin silicon new science and technology company limited)
Powder flowbility analyzer (GTB type, German EWEKA)
Small-sized coating pan (BY300A type, Shanghai Huanghai Sea medicine inspection instrument plant)
Test-type fluid bed (German GLatt company)
Intelligence digestion instrument (ZRS-8C Xing， Radio Factory of Tianjin Univ.)
High performance liquid chromatograph (DIONEX, P680 HpLc Pump, ASI-100 Automated SampLeinjector, Thermostatted CoLumn Compartmeat TCC-100, UVD170U)
Ultraviolet spectrophotometer (SHIMADZU, UV1700)
Optical microscope (JAPAN, OLYMPUS)
1.2 reagents:
Astragalus polysaccharide extract (70%, commercially available prod, lot number is: 080201)
Radix Astragali saponin extract (70%, commercially available prod, lot number is: 080102)
Matrine (98%, according to first matrine of the national drug standards > > of < < State Food and Drug Administration)
Oxymatrine (98%, according to the 16 kurarinone of the national drug standards > > of < < State Food and Drug Administration)
Micropowder silica gel (the extensive Industrial Co., Ltd. in Chengdu, 20080207)
Carboxymethyl starch sodium (the extensive Industrial Co., Ltd. in Chengdu, 20071205)
Microcrystalline Cellulose (101 types, coLorcon)
Eudragit S100, Eudragit E100, Eudragit L100 (German degussa company)
Chitosan (Sigma, 101K4178)
Triethyl citrate (Chengdu Long March chemical reagent factory, 20080105)
Dibutyl phthalate (Chengdu Long March chemical reagent factory, 20071202)
2, the preparation technology of stomach release unit
The preparation of 2.1 Radix Astragali micropills:
2.1.1 the selection of preparation method
Coating pan machinery agitation procedure, two kinds of methods of extrusion-spheronization are carried out to contrast test.
(1) test method
Coating pan machinery agitation procedure, by the ratio of 3: 1: 8, was got Radix Astragali saponin extract, astragalus polysaccharide extract, microcrystalline Cellulose powder that pharmacopeia is sieved (100 orders, 150 μ m) for No. six, mix, put in coating pan, spray into appropriate 50% ethanol, rolling, general ball.
Extrusion-spheronization, in the ratio of 3: 1: 8, was got Radix Astragali saponin extract, astragalus polysaccharide extract, the microcrystalline Cellulose powder of No. six sieves of pharmacopeia (100 orders, 150 μ m), mixed, and appropriate 50% ethanol of take is prepared soft material as wetting agent, extruded, round as a ball.
Measure by the following method the finished product yield of two kinds of preparation methoies, the angle of repose of micropill, friability.
Finished product yield get finished product respectively with No. one, pharmacopeia sieve (10 orders, 2mm), (24 orders, 0.85mm) sieve No. two sieves, and the sample of collection cut size between No. one, No. two sieves, weighs, and according to theoretical yield, calculate finished product yield.
Adopt powder flowbility analyzer angle of repose, to stipulate that conical bottom method measures angle of repose.
Friability is got micropill 10g, adds the bead that 25 diameters are 7mm and puts together the interior rotation of friability instrument 10 minutes, then material is put to No. four sieve (80 orders of pharmacopeia, 250um), jolting 5 minutes, collects and the weighed fine powder amount of passing through sieve, calculates fine powder and accounts for the percentage rate that micropill is heavy.
(2) result of the test
The contrast of the different microsphere and its preparations of table 1
Result shows, agitation procedure is compared with extrusion-spheronization, and molding is difficulty comparatively, and the roundness of gained micropill is poor, friability large, surface is extremely unsmooth, and finished product yield is low.Therefore, select extrusion-spheronization to carry out the preparation of Radix Astragali micropill.
2.1.2 the screening of preparation prescription
(1) impact of preparation prescription on soft material preparation and quality
According to the research to astragalus polysaccharides, Radix Astragali saponin extract physicochemical property, requirement in conjunction with micropill preparation to soft material, work out the prescribed regimen as table 2, get Radix Astragali saponin extract, astragalus polysaccharide extract, microcrystalline Cellulose, micropowder silica gel, cross No. six sieve (100 orders of pharmacopeia, 150 μ m), mix, progressively spray into appropriate 50% ethanol moistening, prepare soft material, the mixing of supplementary material, soft material preparation and quality thereof are investigated, and result of the test is in Table 3.
The different preparation prescription test cards of table 2 (g)
The impact that the different preparation prescriptions of table 3 are prepared soft material
Result shows, the easier mix homogeneously of 6 material of writing out a prescription, and Material Compatibility is better, is difficult for producing segregation phenomenon, is easy to prepare soft material, and gained soft material viscosity is suitable, possesses good plasticity.Therefore, take writes out a prescription 6 carries out computer experiment as basis, further optimize preparation prescription.
(2) impact of preparation prescription on micropill preparation and quality
On preliminary experiment basis, work out the preparation prescription as table 4, by table, get Radix Astragali saponin extract, astragalus polysaccharide extract, microcrystalline Cellulose, micropowder silica gel, carboxymethyl starch sodium powder, with legal system for soft material, under the condition of speed 25Hz, soft material is extruded, under the condition of round as a ball speed 60Hz, rolling 4min, obtains.
The test card of table 4 Radix Astragali micropill prescription screening
To soft material extrude, cutting, round as a ball situation investigate, and by method under 3.2.1.1 item, micropill yield, angle of repose, friability investigated, result of the test is in Table 5.
The result of the test of table 5 Radix Astragali micropill prescription screening
Result shows, the 1 soft material viscosity of writing out a prescription is bigger than normal, extrudes comparatively difficulty after upper machine, and material is larger in extruder internal loss, and soft material plasticity is less than normal, more difficult round as a ball, has part to be small cylindrical, and product yield is not high.Other 2～No. 4 prescription soft material all can be extruded and round as a ball smoothly, and gained micropill roundness, friability are better, and product yield is high, and 2 quality of wherein writing out a prescription are even more ideal, and supplementary product consumption still less, intend selecting prescription 2.
The art for coating of 2.2 Radix Astragali micropills
In Radix Astragali micropill, astragalus polysaccharides, saponin extract critical relative humidity are respectively 63%, 57%, and hygroscopicity is stronger, and the Radix Astragali micropill of preparation is easily moisture absorption also, affects its stability, need to carry out protection against the tide and process.According to preparation design object, Radix Astragali micropill need be intended selecting Eudragit E100 in stomach release fast, and this material has good protection and isolation performance, and can be dissolved in the acid solution below pH5, is usually used in gastric solubleness coating.
3.2.2.1 test method
Coating prescription and coating solution compound method adopt the molten coating prescription of Eudragit E100 alcohol: Eudragit E100 10g; Pulvis Talci 5g; PEG6000 1g; Water 2g; Ethanol 140g.EudragitE100 is dissolved in to about 70g ethanol; PEG6000 is water-soluble; Pulvis Talci added in residue ethanol, by high speed homogenize 20 minutes; PEG6000 solution and Pulvis Talci suspension are added in Eudragit E100 solution, stir evenly, in coating process, continue to stir.
Coating method adopts fluidized-bed coating machine to carry out coating, inlet temperature: 35 ℃, materialization pressure: 1.3～1.45bar, fluidisation pressure: 0.29～0.5bar, feed liquor speed: 19～59, temperature of charge: 28 ℃, delivery temperature: 80 ℃ of left and right, supply gas pressure: 0.68mpa left and right, presses 2mg Eudragit E100/cm
2micropill surface area coating makes micropill weightening finish 5%, obtains.
The quality evaluation of coated micropill get Radix Astragali micropill and coated micropill appropriate, put respectively in weighing botle, bottom set portion fills the glass exsiccator (relative humidity is 75%) of NaCL supersaturated solution, in 25 ℃ of thermostatic drying chambers, preserve, weigh at regular intervals once, calculate the moisture absorption percentage rate of each time.
3.2.2.2 result of the test
Hygroscopic contrast before and after table 6 Radix Astragali coating of pellets
Result shows, micropill substantially no longer moisture absorption after 72 hours after coating, and moisture absorption percentage rate is lower than 1%, and the micropill of coating did not still have obvious moisture absorption at 144 hours, and accumulation moisture absorption percentage rate is 9.86%, after visible coating, micropill moisture pick-up properties obviously reduces.
The contrast of dissolution before and after table 7 Radix Astragali coating of pellets
Result shows, the dissolution in vitro before and after Radix Astragali coating of pellets is except at 10min, there is some difference, and 20min, 30min dissolution no significant difference, show that coating does not make significant difference to the dissolution rate of composition in Radix Astragali micropill.
3, the preparation technology of colon positioning release unit
The preparation of 3.1 sophora root pellets
3.1.1 the selection of preparation method
With reference to the result of study of Radix Astragali micropill, select extrusion-spheronization.
3.1.2 the screening of preparation prescription
(1) screening of diluent
According to the research to Radix Sophorae Flavescentis total alkaloids raw material physicochemical property, selection microcrystalline Cellulose is diluent, intends its consumption to screen.Work out the preparation prescription as table 8, by table, get Radix Sophorae Flavescentis total alkaloids, microcrystalline Cellulose, press 3.2.1.2 item below legal system for micropill, preparation and micropill quality are investigated, and measured micropill yield, angle of repose, friability.Result of the test is in Table 9.
Table 8 diluent screening prescription test card (g)
Table 9 diluent screening test result
Result shows, 4 preparations of writing out a prescription are smooth, and gained micropill quality is better, and product yield is high.Therefore, take and write out a prescription 4 as the further optimization in basis preparation prescription.
(2) screening of disintegrating agent, wetting agent
Selection carboxymethyl starch sodium is disintegrating agent, take water as wetting agent, works out the preparation prescription as table 10, and the consumption of disintegrating agent, wetting agent is screened.By table, get Radix Sophorae Flavescentis total alkaloids, microcrystalline Cellulose, carboxymethyl starch sodium powder, prepare soft material, under the condition of speed 25Hz, soft material is extruded, rolling 4min under the condition of round as a ball speed 60Hz, to soft material extrude, cutting, round as a ball situation investigate, and measure micropill yield, angle of repose, friability.Result of the test is in Table 11.
The test card of table 10 sophora root pellet disintegrating agent, wetting agent screening
Table 11 disintegrating agent, wetting agent screening test result
Result shows, 3, the 4 soft material viscosity of writing out a prescription are bigger than normal, has part to stick phenomenon after upper machine, and it is 1,2 low that micropill yield is write out a prescription, and granularity is not too even.1, No. 2 soft material of writing out a prescription all can be extruded smoothly and be round as a ball, and gained micropill roundness is better, friability is little, and product yield is high, intends selecting prescription 2.
The art for coating research of 3.2 sophora root pellets
Coating design, according to the progress of current colon locating administrated technology, from production application and two angles of basic research, selects three coating schemes to study: 1. pH sensitive single-layer coating: take Eudragit S100 as coating material; 2. the responsive double-layer coatings of pH: internal layer be take Eudragit E100 as coating material, outerly take Eudragit L100 as coating material; 3. pH sensitivity-enzymatic is sent out dual control coating: take chitosan as enzyme control material coating, take Eudragit L100 as enteric coating.
Coating method sprays packaging technique at the bottom of adopting fluid bed, its hydrojet direction is consistent with material movement direction, coating solution with treat that coating material distance is short, drying time is short, can obtain preferably coating uniformity and efficiency, and can, according to material ruuning situation, technological parameter be regulated and controled in good time, guarantee carrying out smoothly of coating, application is comparatively ripe.
Art for coating parameter, in the coating process of each coating scheme, as the case may be, regulates and controls major parameters such as gas flow temperature, throughput, atomizing pressure, hydrojet speed, determines best technological parameter, guarantees coating quality.
3.2.1pH responsive type single coats technique
(1) screening of coating fluid prescription
Reference material service condition
[63]requirement in conjunction with colon positioning release, work out Eudragit S100 coating prescription as table 12, by table, get Eudragit S100, Pulvis Talci, triethyl citrate, dibutyl phthalate, take 95% ethanol as dissolution with solvents, be mixed with the mixed liquor that concentration is about 8% (W/V), after stirring is spent the night, coating solution is crossed 200 mesh sieves and is filtered, 35 ℃ of inlet temperature; Materialization pressure 1.32～1.45bar; Fluidisation pressure 0.29～0.39bar; Feed liquor speed 20～69; 28 ℃ of temperature of charge; 75～80 ℃ of delivery temperatures; Under supply gas pressure 0.56mpa～0.66mpa condition, coating makes to increase weight 25%, takes out coated micropill, adds 1% Pulvis Talci, is dried 2h and get final product in 40 ℃.
The test card of table 12Eudragit S100 coating prescription screening
To the coating process of different coating prescriptions, the outward appearance of coated micropill is investigated, and measures as follows its Release Performance, and result of the test is in Table 13.
Release Performance assay method adopts the second method in second appendix XC dissolution method of < < Chinese Pharmacopoeia > > version in 2005, it is oar method, get micropill 1g, rotating speed 100 ± 1r/min, 37 ± 0.5 ℃ of temperature, release medium is simulated gastric fluid (0.1moL/L hydrochloric acid solution) 900mL, artificial intestinal fluid (phosphate buffer of pH6.8) 1000mL, artificial colonic fluid (phosphate buffer of pH.7.8) 1000mL.Respectively at simulated gastric fluid 2h, artificial intestinal fluid 4h, artificial colonic fluid 2h sampling 5mL (simultaneously adding the dissolution medium of equivalent) crosses 0.46 μ m micro-filtration membrane, obtains.At 250 * 0.46mm, DiamonsiL C18, the phosphate buffer of acetonitrile: pH=3.0 (5: 95), 1mL/min, 40 ℃ of column temperatures, detect under wavelength 205nm chromatographic condition, precision pipettes dissolution fluid 5～20 μ L, sample introduction is measured peak area, and calculates matrine, oxymatrine concentration, with two indexes cubage average accumulated release.
The preparation situation of the different coating prescriptions of table 13 Eudragit S100
Table 14 Eudragit S100 coated micropill cumulative release percentage rate (%)
Result shows, with 1 coating of writing out a prescription, product yield is high, and micropill outward appearance and release in vitro are comparatively desirable.
(2) screening of coating thickness
Eudragit S100 is as enteric coating, and thickness is 30～50um approximately, the about 6mg/cm of polymer overmold amount
2 [63].According to micropill ball, heavy and surface area can calculate the theoretical weightening finish of the coating roughly needing: ball core average diameter 1.4mm, the heavy 2.9mg of average ball;
Micropill surface area estimation: S=4 π r
2=6.1544mm
2;
Coating calculates with polymer: every ball polymer volume 6*6.1544/100=0.3692mg
Theoretical rate of body weight gain=(polymer concentration in amount of polymers/coating)/ball weight=(the 0.3692/0.909)/2.9*100%=14.0% of coating.
The theoretical rate of body weight gain of enteric coating of take is reference, specific requirement in conjunction with colon positioning release, design 15%, 30%, 45% 3 coating thickness level, to screen definite coating prescription 1 by (1) lower method preparation coating solution, and carry out coating, make micropill coating weightening finish 15%, 30%, 45% respectively, take out coated micropill, add 1% Pulvis Talci, in 40 ℃, dry 2h obtains three kinds of coated micropills, and measure micropill release in vitro, result of the test is in Table 15.
The cumulative release percentage rate (%) of the different coating thickness micropills of table 15 Eudragit S100
Result shows, can reduce in a large number its leakage in gastric juice and intestinal fluid, and can in colonic fluid, discharge comparatively rapidly medicine to thicken 30% coated micropill.
3.2.2pH responsive type double-layer coatings technical study
(1) internal layer coating technical study
1. test method
Coating prescription, with reference to Eudragit E100 service condition, in conjunction with the requirement of colon positioning release, adopts the molten coating prescription of Eudragit E100 alcohol: Eudragit E100 10g; Triethyl citrate 1g; 80% ethanol 140mL.
Coating solution compound method is dissolved in about 70mL ethanol by 10g Eudragit E100, and triethyl citrate adds in residue ethanol, and triethyl citrate solution is added in Eudragit E100 solution, after stirring is spent the night, crosses 200 mesh sieves and filters.
Coating method adopts fluidized-bed coating machine to carry out coating, inlet temperature: 35 ℃, and materialization pressure: 1.3～1.45bar, fluidisation pressure: 0.29～0.5bar, feed liquor speed: 19～59, temperature of charge: 28 ℃, delivery temperature: 80 ℃ of left and right, supply gas pressure: 0.68mpa left and right.
Coating thickness Eudragit E100 is about 4mg/cm as the coating thickness of protective separation
2surface area, can calculate according to micropill ball weight and surface area thereof the coating weightening finish roughly needing:
Coating calculates with polymer: every ball polymer volume 4*6.1544/100=0.2462mg;
Theoretical rate of body weight gain=(polymer concentration in amount of polymers/coating)/ball weight=(the 0.2462/0.909)/2.9*100%=9.3% of coating.According to the theoretical rate of body weight gain of coating, 10%, 17%, 25% 3 level of design coating weightening finish is carried out coating, takes out coated micropill, adds 1% Pulvis Talci, and in 40 ℃, dry 2h obtains three kinds of coated micropills, and the release in vitro of coated micropill is investigated.
Release Performance assay method adopts the second method in second appendix XC dissolution method of < < Chinese Pharmacopoeia > > version in 2005, it is oar method, get micropill 1g, phosphate buffer except artificial colonic fluid employing pH4.6, extends to 4h sample time.
2. result of the test
The cumulative release percentage rate (%) of the different coating thickness micropills of table 16 Eudragit E100
Result shows, under the coating prescription of working out and art for coating condition, bag Eudragit E100 clothing layer makes micropill thicken 17%, 25%, and external have a good positioning release medicine performance, considers and select weightening finish 17%.
(2) research of outer art for coating
1. test method
Coating prescription is the service condition for enteric coating with reference to Eudragit L100, works out Eudragit E100 coating prescription: Eudragit L10010g; Triethyl citrate 1g; 95% ethanol 140mL.
Coating solution compound method is dissolved in about 70mL ethanol by Eudragit L100, triethyl citrate adds in residue ethanol, after the stirring of Eudragit L100 alcoholic solution is spent the night, triethyl citrate solution is added in Eudragit E100 solution, stir, cross 200 mesh sieves and filter.
Coating method is got the coated micropill that is surrounded by internal layer clothing film, adopt fluidized-bed coating machine to carry out coating, inlet temperature: 35 ℃, materialization pressure: 1.3～1.45bar, fluidisation pressure: 0.29～0.5bar, feed liquor speed: 19～59, temperature of charge: 28 ℃, delivery temperature: 80 ℃ of left and right, supply gas pressure: under the condition of 0.68mpa left and right, carry out coating.
Coating thickness Eudragit L100 is as enteric coating, and similar to Eudragit S100, thickness is 30～50um approximately, the about 6mg/cm of polymer overmold amount
2[63]can effectively resist the erosion of gastric acid, with reference to result of calculation under 3.4.1.2 item, the theoretical rate of body weight gain of coating is 14.0%, 10%, 15%, 20% 3 level of design coating weightening finish is carried out coating, takes out coated micropill, adds 1% Pulvis Talci, in 40 ℃, dry 2h obtains three kinds of coated micropills, and the release in vitro of coated micropill is investigated.
Release Performance assay method adopts the second method in second appendix XC dissolution method of < < Chinese Pharmacopoeia > > version in 2005, it is oar method, get micropill 1g, rotating speed 100 ± 1r/min, 37 ± 0.5 ℃ of temperature, release medium is simulated gastric fluid (hydrochloric acid solution of pH1.0) 900mL.Respectively at simulated gastric fluid 2h, 4h, 6h sampling 5mL (adding the dissolution medium of equivalent) simultaneously, cross 0.46 μ m micro-filtration membrane, obtain.At 250 * 0.46mm, DiamonsiL C18, the phosphate buffer of acetonitrile: pH=3.0 (5: 95), 1mL/min, 40 ℃ of column temperatures; Detect under wavelength 205nm chromatographic condition, precision pipettes dissolution fluid 5～20 μ L, and sample introduction is measured peak area, and calculates matrine, oxymatrine concentration, with two indexes cubage average accumulated release.
2. result of the test
The cumulative release percentage rate (%) of the different coating thickness micropills of table 17Eudragit L100
Result shows, under the coating prescription of working out and art for coating condition, Eudragit L100 coating increases weight 15% time, and micropill can reach the effect that good antagonism gastric juice infiltrates.
3.2.3pH sensitivity-enzymatic is sent out the research of dual control art for coating
(1) Chitosan Coating technique and micropill performance study
According to the research to chitosan free film performance, intending adopting deacetylation is the coating that 95% chitosan carries out sophora root pellet, and detects the release performance of coated micropill.
1. test method
Coating prescription and coating solution compound method are got chitosan and are added 1% acetate dissolution, are mixed with concentration and are 1% solution, then add the Pulvis Talci of polymer dry weight 100%, the dibutyl phthalate of polymer dry weight 10%, stir spend the night after coating solution cross 200 mesh sieves.
Coating method is got sophora root pellet, adopt fluidized-bed coating machine to carry out coating, inlet temperature: 43～47 ℃, materialization pressure: 1.3～1.5bar, fluidisation pressure: 0.29～0.55bar, feed liquor speed: 15-2535-49, temperature of charge: 28～32 ℃, delivery temperature: 76 ℃ of left and right, supply gas pressure: under the condition of 0.57～0.66mpa left and right, carry out coating.
Coating thickness is according to the result of study to chitosan free film thickness and permeability, thickness is greater than 110 μ m when above, and under artificial intestinal fluid environment, the penetrating speed of matrine is less, therefore, coating makes micropill thicken approximately 150 μ m, and now micropill rate of body weight gain is about 30%.
The basification of Chitosan Coating, with reference to the processing method to chitosan free film, is taken out coated micropill, soaks respectively 10min in 2% aqueous alkali, 30min, and 1h, collects alkaline soak liquid, adds concentrated hydrochloric acid and adjusts pH to 7.0 and be diluted with water to finite concentration, standby.For micropill clear water rapidly washing to neutral, blot immediately surperficial excessive moisture, add 1% Pulvis Talci to mix, standby at 40 degree oven drying 12h.
Alkali treatment Chinese medicine leaks and the mensuration of micropill Release Performance is got three kinds of each 1g of micropill in different alkalization processing times, adopt the second method in second appendix XC dissolution method of < < Chinese Pharmacopoeia > > version in 2005, be that oar method is measured dissolution, rotating speed 100 ± 1r/min, 37 ± 0.5 ℃ of temperature, release medium is artificial intestinal fluid (phosphate buffer of pH6.8) 1000mL.Respectively at artificial intestinal fluid 2h, 4h sampling 5mL (adding the dissolution medium of equivalent) simultaneously, cross 0.46 μ m micro-filtration membrane, obtain.At 250 * 0.46mm, DiamonsiL C18, the phosphate buffer of acetonitrile: pH=3.0 (5: 95), 1mL/min, 40 ℃ of column temperatures; Detect under wavelength 205nm chromatographic condition, precision pipettes each dissolution fluid and alkaline soak liquid 5～20 μ L, and sample introduction is measured peak area, and calculates matrine, oxymatrine concentration, with two indexes cubage average accumulated release or slip.
2. result of the test
The cumulative release degree (%) of table 18 Chitosan Coating micropill
Result shows:
1. coating of pellets film and chitosan free film permeability are basically identical, and in micropill, matrine, oxymatrine can see through Chitosan Coating film swimmingly, in artificial intestinal fluid, exist obviously and leak.
2. in aqueous alkali processing procedure, soak time is longer, and micropill Chinese medicine is more from alkali liquor leakage, and the coated micropill of different soak times Release Performance in artificial intestinal fluid is basically identical, all exists obviously and leaks.
3. result of study prompting, if chitosan is used for to conlon targeting coating, must reduce its permeability to medicine.
(2) Eudragit E100 spacer processes research
Result of study according to (1) item, adopts separately Chitosan Coating, and a large amount of leakages of medicine in intestinal fluid will be inevitable.Therefore, according to the pH environment of intestinal fluid, with reference to pertinent literature, consider to wrap one deck Eudragit E100 contagion gown in chitosan clothing film, to resist intestinal fluid through the etch to the ball heart after chitosan clothing film, reduce the leakage of medicine.
1. test method
Get Eudragit E100 preparation coating solution and carry out coating, coating makes micropill weightening finish 10%.Get chitosan preparation coating solution, and carry out secondary coating to wrapping the coated micropill of Eudragit E100 clothing layer, coating makes micropill weightening finish 30%, with method, above-mentioned double-layer coatings micropill is carried out to basification, obtain Eudragi tE100 and chitosan double-layer coatings micropill, and measure the leakage of basification process Chinese medicine and coated micropill Release Performance.
2. result of the test
The cumulative release degree (%) of table 19 chitosan double-layer coatings micropill
Result shows:
1. this double-layer coatings micropill soaks different time medicine all substantially without leaking in aqueous alkali, in contrast table 3.36, corresponding data can clearly be found out, Eudragit E100 clothing film can effectively resist the etch of alkali liquor to the inner ball heart, and can obviously reduce the leakage of micropill Chinese medicine in intestinal fluid;
2. to double-layer coatings micropill, the leakage in intestinal fluid has a significant effect in aqueous alkali processing, the coated micropill that does not carry out aqueous alkali processing exists obviously and leaks, and along with alkaline soak time lengthening, the leakage of micropill in intestinal fluid slightly reduces, but in alkali liquor, leak also and slightly increase, consider and select alkalization to soak 30min.
(3) Eudragit L100 enteric coating art for coating research
Get Eudragit L100 preparation coating solution, in fluid bed, Eudragit E100 and chitosan double-layer coatings micropill are carried out to coating for the third time, coating makes to increase weight 15%, 20%, 25%, take out coated micropill, add 1% Pulvis Talci, in 40 ℃, dry 2h obtains three kinds of coated micropills, and with method, carries out the investigation of coated micropill release in vitro.The results are shown in Table 20.
The cumulative release percentage rate (%) of the different Eudragit L100 of table 20 coating thickness micropill
Result shows, Eudragit L100 coating increases weight 20% time, and micropill can reach the effect that good antagonism gastric juice infiltrates.
By pharmacodynamics test, prove beneficial effect of the present invention below.
The test of pesticide effectiveness of test example 1 medicine of the present invention
2.5.1 experimental technique
2.5.1.1 experiment grouping
Get SD rat, by body weight, be divided at random 6 groups, normal group, model group, positive group, full side's gavage group, full side's clyster group, Radix Astragali gavage+Radix Sophorae Flavescentis clyster group, every group each 10.
2.5.1.2 the preparation of medicinal liquid
(1) positive group: get sulfasalazine enteric coatel tablets, porphyrize, sieves, porphyrize adds the suspension that normal saline is mixed with 30mg/mL;
(2) full side's gavage group: get astragalus polysaccharide extract, astragaloside extract, matrine, oxymatrine appropriate, add physiological saline solution and be mixed with the mixed solution that concentration is respectively 3.98mg/mL, 1.33mg/mL, 0.51mg/mL, 1.53mg/mL;
(3) full side's clyster group: get said medicine, add normal saline and be mixed with the mixed solution that concentration is respectively 7.96mg/mL, 2.66mg/mL, 1.02mg/mL, 3.06mg/mL;
(4) Radix Astragali gavage+Radix Sophorae Flavescentis clyster group: get astragalus polysaccharide extract, astragaloside extract, add normal saline and be mixed with the mixed solution that concentration is respectively 3.98mg/mL, 1.33mg/mL, separately get matrine, oxymatrine, add the mixed solution that normal saline is mixed with 1.02mg/mL, 3.06mg/mL;
(5) modeling TNBS alcoholic solution: get 5% (w/v) TNBS aqueous solution, add ethanol, be mixed with 50% alcoholic solution of 20mg/mL.Each is organized medicinal liquid and puts into Refrigerator store, stand-by.
2.5.1.3 administration and modeling method
Gavage is pressed 1mL/100g, and coloclysis is pressed 0.5mL/100g administration, positive group gastric infusion, normal group and model group coloclysis give the normal saline of same volume, and every day 1 time, after continuous three days, water 24h is can't help in fasting, the 4th day except normal group, with etherization, by gavage pin by the about 8cm of the light and slow insertion of anus, by 80mg/kg body weight, pour into TNBS solution, hold 30s, make animal keep lying low, naturally clear-headed.Second day rises, and continues administration seven days.
2.5.1.4 evaluation index
Activity, the situation of suffering from diarrhoea, have blood in stool of different time animal after observation modeling, body weight rate of change before and after experiment with computing: (the front body weight of body weight-test before putting to death)/body weight * 100% before testing.
Animal is put to death in femoral artery blood sampling in the 8th day, collects blood in test tube, and the standing natural coagulation of room temperature in 37 ℃ of water-baths, is hatched 20min after shrinking, with the centrifugal 10min of 4000r/min, and separation of serum ,-20 ℃ of preservations, detect for TNF-α.
Get colon, vertical profile is opened, and with ice normal saline, cleans feces, and filter paper suck dry moisture weighs, and calculates the heavy coefficient of intestinal: colon weight/body weight * 100.Then get segment colon and fix with 10% formaldehyde, paraffin embedding, section, HE dyeing, the pathological change of light Microscopic observation colon, its standards of grading are in Table 21.Separately get inflammation and change obvious colon, clean and make homogenate, the centrifugal 15min of 4000r/min, gets supernatant packing, and-20 ℃ of preservations, detect for MPO.
The standards of grading of table 21 colon pathology degree of injury
Statistical method adopts statistic software SPSS 13.0 to carry out one factor analysis of variance.
2.5.2 experimental result
2.5.2.1 outward appearance sign
Within after each treated animal modeling the 2nd day, all occur having blood in stool, diarrhoea situation, there is anorexia, lazy moving phenomenon in most animals, the integral status such as positive group, gavage+clyster group animal activity, diet are obviously better than model group, have blood in stool, diarrhoea degree is all obviously lighter than model group, all the other administration group no significant differences.With normal group comparison, model group rat body weight significantly reduces, and shows that modeling affects large (p < 0.01) to animal diet followed and integrality.Compare with model group, clyster group, gavage+clyster group body weight change have significant difference, can obviously suppress the reduction (p < 0.05,0.01) of the weight of animals, and concrete outcome is in Table 22.
Each treated animal integral status result (x ± S, n=8) of table 22
Note: with normal group comparison,
△p < 0.05,
△ △p < 0.01; With model group comparison,
*p < 0.05,
*p < 0.01.
2.5.2.2 damage is learned by colon
With normal group comparison, the heavy coefficient of model group rat intestine significantly increases (p < 0.01), and rat colon damage is serious, and mucous hyperemia, edema, erosion, have a plurality of ulcer, and damage index has significant difference.Compare with model group, the heavy coefficient of clyster group intestinal significantly reduces, Traumatic Colon is light (p < 0.05), the heavy coefficient of intestinal of gavage+clyster group extremely significantly reduces, Traumatic Colon is light (p < 0.01), show that this medicine can obviously resist the colon hypertrophy that inflammatory damage causes, and the effect of gavage+clyster group is the most obvious, concrete outcome is in Table 23.
The result (x ± S, n=8) of Biao23Ge treated animal colon degree of injury
Note: with normal group comparison,
△p < 0.05,
△ △p < 0.01; With model group comparison,
*p < 0.05,
*p < 0.01.
2.5.2.3TNF-α, MPO testing result
With normal group comparison, the TNF-α of model group, MPO level have utmost point significant difference (p < 0.01).Compare with model group, gavage group only has remarkable reducing effect (p < 0.01) to TNF-α, other administration group all can reduce TNF-α, MPO level (p < 0.01) by significance, wherein remarkable with the effect of gavage+clyster group, show that abnormal immunoreation has better regulating action to this administering mode to body, and the effect that suppresses colon inflammatory reaction is stronger, and concrete outcome is in Table 24.
Table 24 is respectively organized rat blood serum TNF-α, mucous membrane of colon is organized MPO level determination result (x ± S, n=8)
Note: with normal group comparison,
△p < 0.05,
△ △p < 0.01; With model group comparison,
*p < 0.05,
*p < 0.01.
2.5.3 experiment conclusion
By this section research, show:
1. medicine astragalus polysaccharides of the present invention, saponin oral liquor are combined with matrine, oxymatrine coloclysis, can significantly resist the rat colon histopathology damage that trinitro-benzene-sulfonic acid causes, improve that animal is had blood in stool, diarrhoea, anorexia, the lazy outward appearance sign such as moving, and by regulating inflammatory reaction in peripheral blood TNF-alpha levels and colon to play a role.
2. more the peaceful full side's coloclysis administration of intestinal can obviously improve colon's damage, and the inflammatory reaction of colon local organization is had to obvious antagonism, meanwhile, also can obviously resist the abnormal TNF-alpha levels raising in peripheral blood.
3. more the peaceful full side's oral administration human peripheral blood TNF-alpha levels of intestinal has obvious regulating action, and to colon local inflammation, reaction has the trend alleviating.
4. in three kinds of administering modes, the most remarkable with the curative effect that matrine, oxymatrine coloclysis are combined with astragalus polysaccharides, saponin oral liquor, both the whole immunologic function of scalable, can resist colon local inflammatory response again.
The experiment of test example 2 medicine in vivo release of the present invention performance evaluation
1, the research of medicine stomach-colon subrelease capsule of the present invention Release Performance in digestive tract in rats
(1) the in vivo release performance study of stomach release unit
1. test method
Get body weight 220g～250g SD rat, etherization, gets 10 of Radix Astragali coated micropills, and accurately weighed weight adopts gastric intubation administration.Respectively at 15min, 30min, 60min after administration, put to death animal, open abdominal cavity, minute stomach, small intestinal section are got digestive tract, and little intestinal segment starts to ileocecal orifice from pylorus place.Cut intestinal open, observe micropill at intestinal distribution situation and state of appearance (seeing Fig. 2) thereof.
2. result of the test
Distribution and the appearance character (n=3) of table 25 Radix Astragali micropill in rat body
The Release Performance testing result (%) of table 26 Radix Astragali micropill in rat body
Result shows, the stable performance of three batches of Radix Astragali micropill in vivo releases, and all can be complete with interior basic release at 30min, basically identical with experiment in vitro result.
(2) the in vivo release performance study of colon positioning release unit
1. test method
Get body weight 220g～250g SD rat, etherization, gets each 20 of three kinds of Radix Sophorae Flavescentis coated micropills, and accurately weighed weight, adopts gastric intubation administration.Respectively at after administration 1,2,3,4,6,8,10,12,16,20,24hr puts to death animal, opens abdominal cavity, minute stomach, small intestinal, colonic segment are got digestive tract, little intestinal segment starts to ileocecal orifice from pylorus, colonic segment is from ileocecal orifice to rectum end.Cut intestinal open, observe micropill at intestinal distribution situation and state of appearance thereof.Get micropill and measure and wherein remain matrine, oxymatrine concentration by method under 4.2.2 item, and calculate cumulative leaching rate.Separately get not coating element ball of Radix Sophorae Flavescentis, except being set as 0.5,1 sample time, 2hr, all the other the same processing (seeing Fig. 3).
2. result of the test
Distribution and the appearance character (n=3) of table 27 kurarinone ball in rat body
The distributive law (%, n=3) of table 28pH sensitive single-layer type sophora root pellet in rat body
The distribution (n=3) of the responsive double deck type sophora root pellet of table 29pH in rat body
Table 30pH sensitivity-enzymatic is sent out the distribution (n=3) of double-cotrolled type sophora root pellet in rat body
The Release Performance testing result (%, n=3) of table 31 sophora root pellet in rat body
The statistical result that three kinds of micropills are distributed in rat body shows:
1. micropill becomes seriality from the discharge of stomach, and all micropills are discharged approximately and needed 8～10 hours from stomach, discharge half micropill and approximately need 4 hours, and gastric emptying speed exists larger difference between rat individuality; Compare three kinds of micropills, it is relatively slow that pH sensitivity-enzymatic is sent out double-cotrolled type micropill gastric emptying speed, and all the other two kinds of micropill no significant differences.
2. three kinds of situations that micropill does not all exist a large amount of micropills to distribute in each time point small intestinal portion, show that micropill can pass through small intestinal swimmingly, do not have obvious transhipment retardance, and micropill is once gastric emptying, and small intestinal just can be transported to colon comparatively rapidly.
3. three kinds of micropills are approximately just having part to arrive at colon for 3～4 hours after administration, and most of micropill is distributed in colon for 6～10 hours after administration, and wherein pH sensitivity-enzymatic is sent out the time that double-cotrolled type micropill arrives at colon and slightly postponed.
4. in general, if take after administration, within 8 hours, be boundary line, before 8 hours, three kinds of micropills are mainly distributed in the upper digestive tracts such as stomach, small intestinal front end, are mainly distributed in the lower digestive tracts such as distal small intestine or colon portion after 8 hours.
The in vivo release performance of three kinds of micropills of contrast, known in conjunction with its in vivo release tracing analysis:
1. the basic release in 0.5 hour in digestive tract in rats of kurarinone ball is complete, compares with plain ball, and three kinds of coated micropills all have obvious release hesitation.
2. three kinds of coated micropills are in upper gastrointestinal drug release rate and leakage rate, minimum with the responsive double deck type micropill of pH, pH sensitive single-layer type micropill is placed in the middle, and it is maximum that pH sensitivity-enzymatic is sent out double-cotrolled type coated micropill, and within 8 hours, accumulation leakage rate is respectively 11.77%, 17.44%, 27.14%.
3. three kinds of coated micropills lower digestive tract drug release rate and release amount, maximum with pH sensitive single-layer type micropill, the responsive double deck type micropill of pH is placed in the middle, it is minimum that pH sensitivity-enzymatic is sent out double-cotrolled type coated micropill, within 20 hours, preparation is respectively 97.87%, 91.33%, 83.51%, all has obvious colon positioning release feature.
4. in general, three kinds of coated micropills positioning release medicine performance in digestive tract in rats, the most desirable with the responsive double deck type micropill of pH, secondly, it is slightly poor that pH sensitivity-enzymatic is sent out double-cotrolled type micropill to pH sensitive single-layer type micropill.
In sum, drug effect contrast experiment by each component different modes of administration proves, the combination of colon-specific drug delivery system prepared by stomach release system prepared by the Radix Astragali of the present invention or its effective ingredient and Radix Sophorae Flavescentis or its effective ingredient, obviously be better than the full reef knot enteral administration of raw material, two kinds of preparation ways of full side's oral administration, can significantly resist the rat colon histopathology damage that trinitro-benzene-sulfonic acid causes, improving animal has blood in stool, diarrhoea, anorexia, the outward appearance signs such as lazyness is moving, and by regulating inflammatory reaction in peripheral blood TNF-alpha levels and colon to play a role, curative effect is the most remarkable, both the whole immunologic function of scalable, can resist colon local inflammatory response again, by different dosage forms, multiple dosage form comparison in the colon-specific drug delivery system of especially preparing for Radix Sophorae Flavescentis or its effective ingredient, with the best results of the responsive double deck type micropill of pH.
Claims (11)
1. colon-specific drug delivery system prepared by the stomach release system that prepared by the Radix Astragali or its effective ingredient and Radix Sophorae Flavescentis or its effective ingredient purposes in the combination medicine of preparation treatment ulcerative colitis, wherein, described Radix Astragali effective ingredient is: Radix Astragali saponin extract, astragalus polysaccharide extract; The effective ingredient of described Radix Sophorae Flavescentis is Radix Sophorae Flavescentis total alkaloids.
2. purposes according to claim 1, is characterized in that: the described Radix Astragali and the weight proportion of Radix Sophorae Flavescentis are: Radix Astragali 2-6 part, Radix Sophorae Flavescentis 3-7 part.
3. purposes according to claim 2, is characterized in that: the described Radix Astragali and the weight proportion of Radix Sophorae Flavescentis are:
The weight proportion of the Radix Astragali and Radix Sophorae Flavescentis is: 4 parts of the Radixs Astragali, 4 parts of Radix Sophorae Flavescentiss.
4. according to the purposes described in claim 1-3 any one, it is characterized in that: the effective ingredient of the described Radix Astragali is: Radix Astragali saponin extract, astragalus polysaccharide extract; The effective ingredient of described Radix Sophorae Flavescentis is Radix Sophorae Flavescentis total alkaloids, and its weight proportion is:
Radix Astragali saponin extract 10-43 part, astragalus polysaccharide extract 5-40 part, Radix Sophorae Flavescentis total alkaloids 20-70 part.
5. according to the purposes described in claim 1-3 any one, it is characterized in that: the effective ingredient of described Radix Sophorae Flavescentis is matrine and oxymatrine; The weight proportion of Radix Astragali saponin extract, astragalus polysaccharide extract, matrine and oxymatrine is:
Radix Astragali saponin extract 10-43 part, astragalus polysaccharide extract 5-30 part, matrine 5-18 part, oxymatrine 15-52 part.
6. purposes according to claim 5, is characterized in that: the weight proportion of described Radix Astragali saponin extract, astragalus polysaccharide extract, matrine and oxymatrine is:
13.3 parts of astragaloside extracts, 39.8 parts of astragalus polysaccharide extracts, 10.2 parts of matrines, 30.6 parts of oxymatrines.
7. purposes according to claim 4, is characterized in that: the percentage composition that contains astragaloside in described Radix Astragali saponin extract is 5%-10%, and the percentage composition of total saponins is 40%-73.61%; Polyoses content 40%-70% in astragalus polysaccharide extract; The percentage composition that contains total alkaloids in Radix Sophorae Flavescentis total alkaloids is 70%.
8. a combination medicine for the treatment of ulcerative colitis, it is characterized in that, the unit formulation that it comprises different size, for simultaneously, respectively or the colon-specific drug delivery system prepared of the stomach release system prepared of the Radix Astragali of administration successively or its effective ingredient and Radix Sophorae Flavescentis or its effective ingredient, and pharmaceutically acceptable carrier, wherein, described Radix Astragali effective ingredient is: Radix Astragali saponin extract, astragalus polysaccharide extract; The effective ingredient of described Radix Sophorae Flavescentis is Radix Sophorae Flavescentis total alkaloids.
9. the combination medicine for the treatment of ulcerative colitis according to claim 8, it is characterized in that, described stomach release system is to be prepared from micropill by the raw material of following weight proportioning and adjuvant: astragalus polysaccharide extract 199-597 part, Radix Astragali saponin extract 68-204 part, microcrystalline Cellulose 477.5-1432.5 part, micropowder silica gel 93-279 part, carboxymethyl starch sodium 53-159 part, coating 44.5-133.5 part; Described colon-specific drug delivery system is to be prepared from by the raw material of following weight proportioning and adjuvant: Radix Sophorae Flavescentis total alkaloids extract 102-306 part, microcrystalline Cellulose 377.5-1132.5 part, carboxymethyl starch sodium 30.5-91.5 part, coating 306-612 part.
10. combination medicine according to claim 9, is characterized in that: described stomach release system is to be prepared from micropill by the raw material of following weight proportioning and adjuvant: 398 parts of astragalus polysaccharide extracts, 136 parts of Radix Astragali saponin extracts, 955 parts of microcrystalline Cellulose, 186 parts of micropowder silica gels, 106 parts of carboxymethyl starch sodium, 89 parts of coatings; Described colon-specific drug delivery system is to be prepared from by the raw material of following weight proportioning and adjuvant: 204 parts of Radix Sophorae Flavescentis total alkaloids extracts, 755 parts of microcrystalline Cellulose, 61 parts of carboxymethyl starch sodium, coating 306-612 part.
11. according to the combination medicine described in claim 9 or 10, it is characterized in that: the coating described in described stomach release system is Eudragit E100, Eudragit I or other gastric solubleness coating pre-mixing agents; The coating material of described colon-specific drug delivery system is: Eudragit S100, internal layer bag Eudragit E100 outer bag Eudragit L100 or internal layer bag Eudragit E100, middle level involucrum polysaccharide, outer bag Eudragit L100.
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| Title |
|---|
| 中药新复方愈肠宁的组方研究;盛艳梅;《医药导报》;20080331;第27卷(第3期);258-261 * |
| 盛艳梅.中药新复方愈肠宁的组方研究.《医药导报》.2008,第27卷(第3期),258-261. |
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